Supporting Information

 Copyright Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, 2012

Terpenoids are Widespread in Actinomycetes: A

Correlation of Secondary Metabolism and Genome Data
Christian A. Citron,[a] Julia Gleitzmann,[a] Gianfranco Laurenzano,[a] Rdiger Pukall,[b] and
Jeroen S. Dickschat*[a]

cbic_201100641_sm_miscellaneous_information.pdf
 FULL PAPERS
DOI: 10.1002/cbic.200((will be filled in by the editorial staff))

Terpene Biosynthesis

Terpenoids are Widespread in Actinomycetes: A

Correlation of Secondary Metabolism and
Genome Data
Christian A. Citron,[a] Julia Gleitzmann,[a] Gianfranco Laurenzano,[a] Rüdiger
Pukall,[b] and Jeroen S. Dickschat*[a]

Table 1. Bacteria investigated for the production of volatiles and terpene cyclases encoded in their genomes.

ID[a] Strain Geosmin Synthase[b] 2-MIB Synthase[b] Other terpene cyclases[b]

1 Actinosynnema mirum DSM 43827 YP_003098781 – –

2 Catenulispora acidiphila DSM 44928 YP_003116895 YP_003115314 YP_003114277

3 Chitinophaga pinensis DSM 2588 – – YP_003121494

YP_003123612
YP_003123761
YP_003124367

4 Haliangium ochraceum DSM 14365 YP_003265710 – –

5 Herpetosiphon aurantiacus DSM 785 – – YP_001545753

YP_001545754

6 Kitasatospora setae KM-6054 (NBRC 14216) BAJ30389 BAJ32779 BAJ27126

BAJ25873
BAJ31972

7 Kribbella flavida DSM 17836 YP_003382082 – –

8 Ktedonobacter racemifer DSM 44963 – – ZP_06965495

9 Micromonospora olivasterospora NRRL 8178 – BAK26793 –

10 Nocardiopsis dassonvillei subsp. dassonvillei DSM 43111 – YP_003680543 –

11 Rubrobacter xylanophilus DSM 9941 – – YP_643279

12 Saccharopolyspora erythraea NRRL 2338 YP_001105388 YP_001105919 –

YP_001106173
YP_001107098

13 Stackebrandtia nassauensis DSM 44728 YP_003509930 YP_003510780 –

14 Streptomyces albus J1074 ZP_04705018 – ZP_06593440, (+)-epi-isozizaene synthase

15 Streptomyces ambofaciens ATCC 23877 – CAJ89344 –

16 Streptomyces avermitilis MA-4680 NP_823339 – NP_821250, avermitilol synthase

NP_824174, pentalenene synthase
NP_824208, (+)-epi-isozizaene synthase

1
17 Streptomyces clavuligerus ATCC 27064 ZP_06769636 – ZP_05002924
ZP_05002948
ZP_05003209, 1,8-cineol synthase
ZP_05003212
ZP_05004575
ZP_05004823, (-)--cadinene synthase
ZP_05005335
ZP_05005339
ZP_05005402
ZP_05006242, (+)-T-muurolol synthase
ZP_05007964, (3R)-linalool/nerolidol synthase
ZP_05007976
ZP_06775944
ZP_08219819
ZP_08219821

18 Streptomyces coelicolor A3(2) NP_630182 NP_733742 NP_629369, (+)-epi-isozizaene synthase

19 Streptomyces flavogriseus ATCC 33331 ADW07414 ADW07061 ADW03055

ADW05938
ADW06796, (+)-1-epi-cubenol synthase

20 Streptomyces ghanaensis ATCC 14672 ZP_04685148 – ZP_06576746, (+)-epi-isozizaene synthase

21 Streptomyces griseoflavus Tü 4000 ZP_07309957 – ZP_07310844, (+)-epi-isozizaene synthase

22 Streptomyces griseus subsp. griseus NBRC 13350 YP_001828351 YP_001822781 YP_001823591, (+)-caryolan-1-ol synthase
YP_001827577, (+)-1-epi-cubenol synthase

23 “Streptomyces lasaliensis” NRRL 3382 – BAI77523 –

24 Streptomyces lividans TK 24 ZP_05522837 ZP_05521425 ZP_06528571, (+)-epi-isozizaene synthase

ZP_05521426 ZP_06533634

25 Streptomyces peucetius subsp. caesius ATCC 27952 ABY50951 – –

26 Streptomyces pristinaespiralis ATCC 25486 ZP_06913794 – ZP_06911744

ZP_06913376

27 Streptomyces filamentosus NRRL 15998 – – ZP_06582730, (+)-1-epi-cubenol synthase

ZP_06587258, (+)-caryolan-1-ol synthase

28 Streptomyces sp. Tü 6071 ZP_08453284 – ZP_08452581, (-)-epi-zizaene synthase

29 Streptomyces scabiei 87.22 YP_003487693 YP_003486275 YP_003492893

YP_003493696

30 Streptomyces sviceus ATCC 29083 ZP_06920565 – ZP_06919672, (+)-epi-isozizaene synthase

31 Streptomyces venezuelae ATCC 10712 CCA53556 CCA60397 CCA53839

32 Streptomyces violaceusniger Tü 4113 ZP_07608000 ZP_07604855 ZP_07605120

33 Streptomyces viridochromogenes DSM 40736 ZP_07307300 ZP_07301463 ZP_07302078

ZP_07306383, (+)-epi-isozizaene synthase
ZP_07308339

34 Streptosporangium roseum DSM 43021 YP_003335875 – YP_003338321

YP_003342315

35 Thermomonospora curvata DSM 43183 – – –

[a] Strain identification number used in Table 2. [b] Genebank accession number. A minus sign indicates that no orthologous gene is encoded in the genome of the
respective organism.

2
Table 2. Volatile terpenoids from bacteria.

[a] [b] [c] [d] [e]

Compound I I (Lit.) Identification Found in the strains
[1]
-Thujene (82) 925 924 ms, ri 26
[1]
-Pinene (65) 933 932 ms, ri, std 1, 3, 5, 9, 26, 31, 34
[1]
Camphene 947 946 ms, ri 1
[1]
-Pinene (66) 975 974 ms, ri, std 1, 5, 9, 31

2-Methylenebornane (64) 978 ms 6, 10, 19, 22, 24, 31, 33

[1]
-3-Carene 1008 1008 ms, ri 1, 3, 5

2-Methyl-2-bornene (63) 1015 ms 6, 9, 10, 12, 19, 22, 23, 24, 29, 31, 32, 33
[1]
Limonene (81) 1023 1024 ms, ri, std 1, 2, 3, 4, 5, 7, 12, 13, 15, 16, 17, 20, 21, 25, 26, 30, 32, 33, 34
[1]
1,8-Cineole (76) 1026 1026 ms, ri, std 4, 12, 17, 22, 32
[1]
cis-Linalool oxide 1072 1067 ms, ri 2, 20, 28, 30, 34
[1]
Dihydromyrcenol 1073 1069 ms, ri 1, 2, 3, 5, 12, 13, 16, 17, 22, 25, 33, 34
[1]
trans-Linalool oxide 1086 1084 ms, ri 12, 17, 28, 30
[1]
Linalool (77) 1098 1095 ms, ri 1, 6, 12, 15, 17, 19, 23, 27, 28

Tetrahydrolinalool 1098 1098 ms, ri 30

[1]
Nopinone 1135 1135 ms, ri 38
[1]
Camphor 1142 1141 ms, ri, std 1, 6, 12, 21, 25, 26
[1]
Isoborneol 1155 1155 ms, ri 12, 22
[1]
-Terpineol 1164 1162 ms, ri 17
[1]
Borneol 1164 1165 ms, ri 22
[1]
Isomenthol 1174 1179 ms, ri 12
[1]
2-Methylisoborneol (2) 1177 1178 ms, ri, std 6, 9, 10, 12, 15, 18, 19, 22, 23, 24, 29, 31, 32, 33

3-Oxocineol 1179 ms 17
[1]
-Terpineol 1188 1186 ms, ri 15, 17, 27

(8S,9R,10S)-8,10-Dimethyl-1-octalin (4) 1224 ms, std 1, 4, 6, 12, 14, 15, 16, 18, 19, 20, 21, 23, 24, 26, 28, 29, 30, 31, 32, 33

Citronellol 1226 1223[1] ms, ri 12

(8S,10R)-8,10-Dimethyl-1(9)-octalin (39) 1233 ms, std 1, 4, 6, 12, 14, 15, 16, 18, 19, 20, 21, 23, 24, 26, 28, 29, 30, 31, 32, 33

Carvon 1242 1239[1] ms, ri 17

(10R)-8,10-Dimethyl-8-octalin (43) 1245 ms, std 1, 12, 14, 15, 16, 21, 30
[1]
Geraniol 1254 1249 ms, ri, std 12
[1]
Cogeijerene (44) 1283 1283 ms, ri 15, 21, 30
[1]
Silphiperfol-5-ene (115) 1325 1326 ms, ri 16, 32
[1]
-Elemene 1335 1335 ms, ri 21, 25
[2]
Bicycloelemene (119) 1336 1338 ms, ri 12, 14, 19, 27, 28, 30, 33
[2]
Pentalenene (12) 1339 1343 ms, ri 16
[1]
-Cubebene (96) 1348 1348 ms, ri 14, 19, 21, 22, 27, 30, 34
[2]
African-1-ene (107) 1354 1356 ms, ri 32
[2]
Clovene 1360 1365 ms, ri 27
[2]
Dehydrogeosmin (47) 1367 1362 ms, ri 15, 21, 26, 30
[1]
-Ylangene 1371 1373 ms, ri 14, 21, 30

-Copaene (97) 1374 1374[1] ms, ri 14, 19, 21, 22, 27, 30, 34
[1]
Silphiperfol-6-ene (114) 1377 1377 ms, ri 16, 32

3
 [2]
African-3-ene (108) 1387 1391 ms, ri 17, 32
[1]
Isolongifolene 1387 1389 ms, ri 12
[1]
-Cubebene (32) 1387 1387 ms, ri 1, 12, 14, 19, 21, 26, 27, 28, 30, 31, 34
[1]
-Bourbonene (112) 1387 1387 ms, ri 23
[1]
-Elemene (120) 1389 1389 ms, ri 1, 12, 13, 14, 15, 16, 17, 18, 19, 21, 23, 26, 27, 28, 30, 31, 32, 33
[1]
Geosmin (1) 1401 1399 ms, ri, std 1, 4, 6, 12, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 28, 29, 30,

31, 32, 33, 34

[2]
African-3(15)-ene 1403 1400 ms, ri 32
[1]
Longifolene (122) 1403 1407 ms, ri 17
[3]
6,10-Dimethylundecan-2-one 1406 1408 ms, ri 2, 3, 11, 15, 16, 19, 20, 25, 26, 27
[1]
-Barbatene (109) 1408 1407 ms, ri 17
[1]
-Gurjunene (106) 1409 1409 ms, ri 22, 27

-Cedrene (105) 1412 1410[1] ms, ri, std 14, 15, 18, 21, 31
[1]
cis--Bergamotene 1413 1411 ms, ri 17
[2]
Tritomarene (111) 1413 1416 ms, ri 19, 22, 27
[1]
-Funebrene (104) 1413 1413 ms, ri 8, 14, 21, 28

Aristolene (102) 1418 1423[2] ms, ri 19, 22

[1]
(E)--Caryophyllene (117) 1418 1417 ms, ri 17, 27, 32
[1]
-Ylangene (30) 1419 1419 ms, ri 1, 12, 14, 16, 21, 23, 28, 29, 30, 31, 33
[1]
-Cedrene (62) 1420 1419 ms, ri, std 8, 13, 14, 18, 21, 28

Bourbon-11-ene (113) 1421 1424[2] ms, ri 19, 22, 27

Opposita-4(15),11-diene 1426 1424[2] ms, ri 30

cis-Thujopsene 1429 1429[1] ms, ri 17

-Copaene (31) 1430 1430[1] ms, ri 1, 12, 14, 16, 19, 21, 23, 30, 31, 34

-Gurjunene/Calarene (103) 1430 1431[1] ms, ri 19, 22, 27

[1]
-Elemene (121) 1432 1434 ms, ri 25, 26, 27
[1]
trans--Bergamotene 1434 1432 ms, ri 14

Isobazzanene 1435 1436[1] ms, ri 17

[1]
-Barbatene (110) 1441 1440 ms, ri 17
[2]
Selina-5,11-diene (99) 1442 1444 ms, ri 19, 22, 27, 30
[2]
Sesquisabinene B 1442 1446 ms, ri 14, 18, 21
[2]
Isogermacrene D 1443 1445 ms, ri 12, 30, 31

epi-Isozizaene (9) 1444 ms 14, 15, 16, 18, 20, 21, 23, 24, 28, 33
[1]
-Himachalene 1447 1449 ms, ri 17
[1]
Cadina-3,5-diene (91) 1451 1451 ms, ri 19, 27, 34
[2]
Cadina-4,11-diene 1449 1446 ms, ri 13
[1]
-Humulene (116) 1453 1452 ms, ri, std 16, 22, 27

epi-Prezizaene (70) 1453 14, 18, 28

[1]
Geranyl acetone 1453 1453 ms, ri, std 2, 3, 4, 5, 8, 11, 12, 15, 16, 17, 21, 26, 32, 33, 34
[2]
-Muurolene (90) 1454 1455 ms, ri 12, 21, 30
[1]
(E)--Farnesene 1455 1454 ms, ri 17

epi-Zizaene (71) 1460 14, 15, 18, 20, 21, 28

[1]
-Acoradiene (58) 1462 1464 ms, ri 13, 17

4
 [4]
Muurola-4(15),5-diene (93) 1463 1468 ms 19, 27
[2]
6,11-Epoxyisodaucane (24) 1464 1463 ms 1, 6, 12, 14, 15, 16, 18, 19, 21, 23, 24, 26, 28, 29, 30, 31, 33

(1R*,6S*,10S*)-6,10- 1469 ms, std 12, 14, 30

Dimethylbicyclo[4.4.0]decan-3-one (45)
[1]
trans-Cadina-1(6),4-diene (86) 1472 1475 ms, ri 19, 27, 34
[1]
-Neocallitropsene 1473 1474 ms, ri 8, 14
[2]
Selina-4,11-diene 1474 1475 ms, ri 12
[1]
-Chamigrene 1474 1476 ms, ri 17

10-epi--Acoradiene 1476 1474[1] ms, ri 28

[1]
-Muurolene (89) 1477 1478 ms, ri 19, 21, 22, 27, 30, 34
[1]
-Curcumene (54) 1478 1481 ms, ri 2
[2]
cis-4,10-epoxy-Amorphane (98) 1478 1481 ms, ri 17
[2]
Amorpha-4,7(11)-diene 1480 1479 ms, ri 17
[1]
Germacrene D (5) 1480 1480 ms, ri 1, 6, 12, 14, 15, 16, 18, 19, 21, 23, 24, 26, 28, 29, 30, 31, 33
[1]
-Curcumene (59) 1481 1479 ms, ri 2, 17, 32
[1]
-Amorphene (95) 1483 1483 ms, ri 33
[2]
Isolepidozene (20) 1484 1483 ms, ri 12, 28
[1]
(E)--Ionone 1485 1487 ms, ri 11

-Selinene (100) 1487 1489[1] ms, ri 19, 21

[1]
Bicyclosesquiphellandrene (92) 1492 1493 ms, ri 12, 19, 21, 27, 30, 34
[2]
-Selinene 1495 1498 ms, ri 19, 21
[1]
Bicyclogermacrene (16) 1497 1500 ms 31
[1]
epi-Zonarene 1499 1501 ms, ri 21, 27
[1]
-Muurolene (94) 1500 1500 ms, ri 19, 22, 26, 27, 34

(Z)--Bisabolene (51) 1503 1506[1] ms, ri 7, 13

Cuparene 1504 1504[1] ms, ri 17

[1]
Isodihydroagarofuran (36) 1503 1503 ms, ri, std 1, 6, 12, 14, 15, 16, 18, 19, 21, 23, 24, 26, 28, 29, 30, 31, 33, 34
[1]
-Bisabolene (50) 1506 1505 ms, ri 2, 7, 13, 15, 17, 19
[1]
-Curcumene (55) 1510 1514 ms, ri 2, 17

-Cadinene (83) 1513 1513[1] ms, ri 3, 14, 17, 19, 21, 22, 27, 30

-Alaskene (57) 1514 1512[1] ms, ri 7, 8, 13

[1]
Cubebol 1514 1514 ms, ri 27
[1]
(Z)--Bisabolene 1513 1514 ms, ri 14, 18
[1]
-Sesquiphellandrene 1520 1521 ms, ri 14

trans-Calamenene (87) 1521 1521[1] ms, ri 12, 14, 19, 22, 27

[1]
-Cadinene (79) 1522 1522 ms, ri 14, 17, 19, 21, 27, 34
[1]
Zonarene (88) 1525 1528 ms, ri 19, 22, 27
[5]
Dihydroactinidiolide 1530 1525 ms, ri 11, 35
[1]
Cadina-1,4-diene (85) 1532 1533 ms, ri 19, 22, 27, 34
[1]
-Cadinene (84) 1536 1537 ms, ri 14, 17, 19, 21, 22, 27, 30
[2]
Selina-4(15),7(11)-diene (101) 1537 1534 ms, ri 25, 26
[1]
Selina-3,7(11)-diene 1541 1545 ms, ri 26
[1]
-Calacorene 1544 1544 ms, ri 14, 19, 27

5
 [1]
-Agarofuran (40) 1544 1548 ms, ri 19
[6]
(E)--Bisabolene (52) 1544 1540 ms, ri 7, 13

(6S*,10S*)-6,10-Dimethylbicyclo[4.4.0]dec-1- 1551 ms, std 30

en-3-one (46)
[1]
Germacrene B 1561 1559 ms, ri 26
[1]
(E)-Nerolidol (78) 1561 1561 ms, ri 2, 11, 17
[1]
-Calacorene 1563 1564 ms, ri 19, 21

Caryolan-1-ol (74) 1564 ms 22, 27

[1]
Germacrene D-4-ol (118) 1574 1574 ms, ri 19

8-epi-Rosifoliol (28) 1587 8, 12, 14, 15, 21, 28, 30, 33

(4S)-Albaflavenol (10a) 1616 ms 14, 18, 20

(4R)-Albaflavenol (10b) 1626 ms 14, 15, 18, 20, 21

[1]
epi-Cubenol (75) 1627 1627 ms, ri 19, 22, 27
[1]
-Acorenol 1631 1632 ms, ri 28

T-Muurolol (80) 1641 1640[1] ms 17, 27, 34

(1(10)E,5E)-Germacradien-11-ol (3) 1642 ms, std 1, 4, 6, 12, 14, 15, 16, 17, 18, 19, 20, 21, 23, 24, 26, 28, 29, 30, 31, 33

-Cadinol 1654 1652[1] ms, ri 19, 27

7-epi--Eudesmol (35) 1657 1662[1] ms, ri 15, 19

4,5-Epoxy-2-epi-zizaan-6-ol (11) 1675 ms 15, 18, 21, 23, 24, 33

[1]
Cadalene 1675 1675 ms, ri 19

Albaflavenone (8) 1690 ms 14, 15, 18, 20, 21, 23, 28, 33

(Z)-Nuciferal (60) 1731 1727 ms, ri 2

Mintsulfide 1741 1740[1] ms, ri 21

6,10,14-Trimethylpentadecan-2-one 1842 ms, std 11, 15, 18, 26

Cembrene (125) 1934 1937[1] ms, ri 11

Isocembrene 1947 1951[2] ms, ri 11

(3Z)-Cembrene A (124) 1962 1965[1] ms, ri 11

[7]
Cembrene C (123) 2006 2002 ms, ri 11

[a] Unidentified compounds, artifacts, and medium compounds are not mentioned. [b] Arithmetic retention indices according to van den Dool and Kratz [Ref]. [c]
Literature data of arithmetic retention indices determined on the same (HP-5 MS) or similar GC column. [d] Compound identification based on comparison of the
mass spectrum to a data base spectrum (ms), a retention index matching literature data within a range of ±5 index points (ri), or direct comparison to a synthetic or
commercially available standard (std). [e] Numbers refer to the strain identification numbers given in Table 1.

6
Table 3. Growth conditions of investigated bacteria.

[a] [b] [b]

ID Strain Growth temperature Medium 1 Medium 2

1 Actinosynnema mirum DSM 43827 28°C 65 (pH 7.2) –

2 Catenulispora acidiphila DSM 44928 28°C 65 (pH 5.5) SFM (pH 5.5)

3 Chitinophaga pinensis DSM 2588 22°C 67 (pH 7.2) –

4 Haliangium ochraceum DSM 14365 28°C 958 (pH 7.5) –

5 Herpetosiphon aurantiacus DSM 785 30°C 67 (pH 7.2) –

6 Kitasatospora setae KM-6054 (NBRC 14216) 28°C 214 (pH 7.2) SFM (pH 7.2)

7 Kribbella flavida DSM 17836 28°C 830 (pH 7.2) SFM (pH 7.2)

8 Ktedonobacter racemifer DSM 44963 28°C 65 (pH 6.0) SFM (pH 6.0)

9 Micromonospora olivasterospora NRRL 8178 28°C 65 (pH 7.2) SFM (pH 7.2)

10 Nocardiopsis dassonvillei subsp. dassonvillei DSM 43111 28°C 65 (pH 7.2) –

11 Rubrobacter xylanophilus DSM 9941 50°C 878 (pH 7.2) –

12 Saccharopolyspora erythraea NRRL 2338 28°C 65 (pH 7.2) SFM (pH 7.2)

13 Stackebrandtia nassauensis DSM 44728 28°C 554 (pH 7.2) SFM (pH 7.2)

14 Streptomyces albus J1074 28°C 65 (pH 7.2) SFM (pH 7.2)

15 Streptomyces ambofaciens ATCC 23877 28°C 65 (pH 7.2) SFM (pH 7.2)

16 Streptomyces avermitilis MA-4680 28°C 425 (pH 7.2) SFM (pH 7.2)

17 Streptomyces clavuligerus ATCC 27064 28°C 65 (pH 7.2) SFM (pH 7.2)

18 Streptomyces coelicolor A3(2) 28°C 65 (pH 7.2) SFM (pH 7.2)

19 Streptomyces flavogriseus ATCC 33331 28°C 65 (pH 7.2) SFM (pH 7.2)

20 Streptomyces ghanaensis ATCC 14672 28°C 65 (pH 7.2) SFM (pH 7.2)

21 Streptomyces griseoflavus Tü 4000 28°C 65 (pH 7.2) SFM (pH 7.2)

22 Streptomyces griseus subsp. griseus NBRC 13350 28°C 228 (pH 7.3) SFM (pH 7.2)

23 “Streptomyces lasaliensis” NRRL 3382 28°C 425 (pH 7.2) SFM (pH 7.2)

24 Streptomyces lividans TK 24 28°C 65 (pH 7.2) SFM (pH 7.2)

25 Streptomyces peucetius subsp. caesius ATCC 27952 28°C 65 (pH 7.2) SFM (pH 7.2)

26 Streptomyces pristinaespiralis ATCC 25486 28°C 65 (pH 7.2) SFM (pH 7.2)

27 Streptomyces filamentosus NRRL 15998 28°C 65 (pH 7.2) SFM (pH 7.2)

28 Streptomyces sp. Tü 6071 28°C 65 (pH 7.2) SFM (pH 7.2)

29 Streptomyces scabiei 87.22 28°C 65 (pH 7.2) SFM (pH 7.2)

30 Streptomyces sviceus ATCC 29083 28°C 65 (pH 7.2) SFM (pH 7.2)

31 Streptomyces venezuelae ATCC 10712 28°C 65 (pH 7.2) SFM (pH 7.2)

32 Streptomyces violaceusniger Tü 4113 28°C 65 (pH 7.2) SFM (pH 7.2)

33 Streptomyces viridochromogenes DSM 40736 28°C 65 (pH 7.2) SFM (pH 7.2)

34 Streptosporangium roseum DSM 43021 28°C 65 (pH 7.2) –

35 Thermomonospora curvata DSM 43183 45°C 550 (pH 7.2) –

[a] Strain identification number used in Table 2. [b] For medium compositions see below.

7
Medium compositions

Medium ingredients are given per 1 L of distilled water. For agar plates 15 g of agar (Carl Roth GmbH) were added. The pH was
adjusted according to Table 3.

Medium 65

4.0 g Glucose
4.0 g Yeast extract
10.0 g Malt extract
2.0 g CaCO3 (only for agar plates, do not add for liquid cultures)

Medium 67

3.0 g Casitone
1.36 g CaCl2 x 2 H2O
1.0 g Yeast extract

Medium 214

4.0 g Glucose
4.0 g Yeast extract
10.0 g Malt extract
2.0 g CaCO3 (only for agar plates, do not add for liquid cultures)
20.0 g Starch

Medium 228

1.0 g Yeast extract

1.0 g Beef extract
2.0 g NZ Amine
10.0 g Glucose

Medium 425

10.0 g Oat flakes

10.0 g Oatmeal

Medium 550

48 g Czapek Dox Agar

2.0 g Yeast extract
6.1 g Casamino acids
0.02 g Tryptophan

Medium 554

10.0 g Glucose
20.0 g Starch, soluble
5.0 g Yeast extract
5.0 g NZ Amine
1.0 g CaCO3

8
Medium 830

0.5 g Yeast extract

0.5 g Proteose peptone
0.5 g Casamino acids
0.5 g Glucose
0.5 g Starch, soluble
0.3 g Sodium pyruvate
0.3 g K2HPO4
0.05 g MgSO4 x 7 H2O

Medium 878

1.0 g Yeast extract

1.0 g Tryptone
0.1 g Nitrilotriacetic acid
0.04 g CaSO4 x 2 H2O
0.2 g MgCl2 x 6 H2O
0.5 mL 0.01 M Fe(III)-citrate
0.5 mL Trace element solution
100 mL Phosphate buffer

Trace element solution

0.5 mL H2SO4
2.28 g MnSO4 x H2O
0.5 g ZnSO4 x 7 H2O
0.5 g H3BO3
25 mg CuSO4 x 5 H2O
25 mg Na2MoO4 x 2 H2O
45 mg CoCl2 x 2 H2O
1L Distilled water

Phosphate buffer

5.44 g KH2PO4
43 g Na2HPO4
1L Distilled water, adjust to pH 7.2

Medium 958

20.0 g NaCl
2.5 g Yeast cell paste (baker’s yeast, washed in distilled water, wet weight)
0.5 mg Cyanocobalamine (vitamin B12)
1L Sea water salt solution (instead of distilled water)

Sea water salt solution

0.01 g Fe(III)-citrate
8.0 g MgSO4 x 7 H2O
1.0 g CaCl2 x 2 H2O
0.5 g KCl
0.16 g NaHCO3
0.02 g H3BO3
0.08 g KBr
0.03 g SrCl2 x 6 H2O
0.01 g Disodium--glycerophosphate
1 mL Trace elements solution
1L Distilled water

9
SFM (soja flour medium)

20.0 g Mannitol
20.0 g Soja flour

10
 Figure 1. Alignment of geosmin synthases.

Position 1 11 21 31 41 51 61
YP_003098781 (DSM 43827) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
AEK39836 (S699) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
YP_003763541 (U32) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - M A D T A A G W
AEA03338 (CHAB 1432) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
AEA03341 (CHAB 2155) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
YP_716636 (ACN14a) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
YP_483306 (Ccl3) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
YP_001509819 (EAN1pec) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
YP_004017381 (Eul1c) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
YP_003265710 (DSM 14365) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
BAJ30389 (KM-6054) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
YP_634376 (DK 1622) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
YP_001866236 (PCC 73102) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ZP_07114089 (PCC 6056) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ABU93239 (P2r) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ABU93238 (P2r) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
YP_001105388 (NRRL 2338) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
YP_001107098 (NRRL 2338) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
YP_001106173 (NRRL 2338) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
YP_001612078 (So ce 56) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ZP_01460669 (DW 4/3-1) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
YP_003950745 (DW 4/3-1) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ZP_04705018 (J1074) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
NP_823339 (MA-4680) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ADI05189 (BCW-1) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
CCB75658 (NRRL 8057) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ZP_08240572 (XylebKG-1) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ZP_06769636 (ATCC 27064) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
NP_630182 (A3(2)) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ADW07414 (ATCC 33331) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ZP_04685148 (ATCC 14762) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ZP_08289513 (M045) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ZP_07309957 (Tü 4000) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
YP_001828351 (NBRC 13350) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ZP_07296488 (ATCC 53653) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ZP_07293917 (ATCC 53653) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ZP_05522837 (TK 24) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ABY50951 (ATCC 27952) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ZP_06913794 (ATCC 25486) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
YP_003487693 (87.22) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ZP_07276967 (AA4) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ZP_06273105 (ACTE) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ZP_07289987 (C) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ZP_05001875 (Mg1) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ZP_07976693 (SA3_actG) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ZP_07273435 (SPB78) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ZP_08453284 (Tü 6071) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ZP_06920565 (ATCC 29083) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - M G
CCA53556 (ATCC 10712) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ZP_07608000 (Tü 4113) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ZP_07307300 (DSM 40736) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
YP_003335875 (DSM 43021) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
YP_003382082 (DSM 17836) M R A P Q P P G A A A R L V S H V G R D A T D E P G G L T K G G S P P T G R P T P S T G Q S T L R T G Q P T S N A R Q P A P S R E
YP_003509930 (DSM 44728) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
YP_003116895 (DSM 44928) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
Highly conserved residues are shown in red, deviations from otherwise highly conserved residues are shown in dark red. The strictly conserved motifs are shown in boxes. Added nucleotides of corrected sequences are shown in bold.

11
 Figure 1. Alignment of geosmin synthases.

71 81 91 101 111 121 131

YP_003098781 (DSM 43827)
AEK39836 (S699)
YP_003763541 (U32)
AEA03338 (CHAB 1432)
AEA03341 (CHAB 2155)
YP_716636 (ACN14a)
YP_483306 (Ccl3)
YP_001509819 (EAN1pec)
YP_004017381 (Eul1c)
YP_003265710 (DSM 14365)
BAJ30389 (KM-6054)
YP_634376 (DK 1622)
YP_001866236 (PCC 73102)
ZP_07114089 (PCC 6056)
ABU93239 (P2r)
ABU93238 (P2r)
YP_001105388 (NRRL 2338)
YP_001107098 (NRRL 2338)
YP_001106173 (NRRL 2338)
YP_001612078 (So ce 56)
ZP_01460669 (DW 4/3-1)
YP_003950745 (DW 4/3-1)
ZP_04705018 (J1074)
NP_823339 (MA-4680)
ADI05189 (BCW-1)
CCB75658 (NRRL 8057)
ZP_08240572 (XylebKG-1)
ZP_06769636 (ATCC 27064)
NP_630182 (A3(2))
ADW07414 (ATCC 33331)
ZP_04685148 (ATCC 14762)
ZP_08289513 (M045)
ZP_07309957 (Tü 4000)
YP_001828351 (NBRC 13350)
ZP_07296488 (ATCC 53653)
ZP_07293917 (ATCC 53653)
ZP_05522837 (TK 24)
ABY50951 (ATCC 27952)
ZP_06913794 (ATCC 25486)
YP_003487693 (87.22)
ZP_07276967 (AA4)
ZP_06273105 (ACTE)
ZP_07289987 (C)
ZP_05001875 (Mg1)
ZP_07976693 (SA3_actG)
ZP_07273435 (SPB78)
ZP_08453284 (Tü 6071)
ZP_06920565 (ATCC 29083)
CCA53556 (ATCC 10712)
ZP_07608000 (Tü 4113)
ZP_07307300 (DSM 40736)
YP_003335875 (DSM 43021)
YP_003382082 (DSM 17836)
YP_003509930 (DSM 44728)
YP_003116895 (DSM 44928)
Highly conserved residues are shown in red, deviations from otherwise highly conserved residues are shown in dark red. The strictly conserved motifs are shown in boxes. Added nucleotides of corrected sequences are shown in bold.
- - - - - M P Q P F Q L P E F Y M P Y P A R L N P N L E Q A R A H S K A W A R S M D M I D V P Q H G T - - - - V V W D E S D L D S
- - - - M P E Q P F V L P E F Y L P Y P A R L N P H L D R A R E H S K A W A R E H D M I D V P Q H G T - - - - V I W T E H D L D S
L P R G V P E Q P F V L P E F Y L P Y P A R L N P H L D R A R E H S K A W A R E H D M I D V P Q H G T - - - - V I W T E H D L D S
- - - - - - M Q P F K L P A F Y M P W P A R L N P N L E A A R V H S K A W A Y E M G I L G S K E E S Q - - G E P I W D E R K F D A
- - - - - - M Q P F K L P A F Y M P W P A R L N P N L E A A R V H S K A W A Y E M G I L G S K E E S Q - - G E P I W D E R K F D A
- - - - - - M Q P F T L P E F Y V P Y P A R L S P H L E Q A R E H S R E W A R A M E M I D T P Q H G I - - - - A I W T E R D L D A
- - - - - - M Q P F T L P E F Y V P Y P A R L N P N L E Q A R V H S R A W A D E M E M I D S P Q H G T - - - - A I W T E A D F D A
- - - - - - M Q P F T L P E F Y L P Y P P R L N P N L E H A R V H S R A W A G E M E M I D V P Q D G V - - - - A I W S G Q D F D S
- - - - - - M K P F T L P R F Y V P Y P A R L S P H L D A A R T H S R A W A A E M E M I G P A P G G E - - - - V I W S E Q D F D A
- - - - - M S H P F Q L P E F Y V P Y P A R L N P N L E G A R V H S K A W A F E M G M L G S E D D A A S G G E P I W T E R K L D A
- - - - - M A Q P F E L P D F Y V P Y P A R I N P H Y E R A R V H T K E W A R G F G M L E - - - - G S - - - - G V W E E H D L D S
M S T A K N K Q P F E L P D F Y V P W P A R L N P N L E G A R V H S K A W A R E L G I I G R P K D G S - - A P E I W S E A K F D A
- - - - - - M Q P F E L P E F Y M P W P A R L N P N L E A A R S H S K A W A Y Q M G I L G S K E E A E - - S S V I W D E R T F D A
- - - - - - M Q P F K L P D F Y M P W P A R L N P N L E A A R V H S K A W A Y E M G I L A S E E E S Q - - D E P I W D E R T F D A
- - - - - - M Q P F K L P D F Y M P W P A R L N P N L E A A R V H S K A W A Y D M G I L G S K E E A N - - G E P I W D E R K F D A
- - - - - - M Q P F K L P D F Y M P W P A R L N P N L E A A R V H S K A W A Y E M G I L G S K E E A N - - G E P I W D E R K F D A
- - - - - - M Q P F Q Q P E F Y M P Y P A R L N P N L E R A R E H S K A W A C A M D M I D V P Q E G T - - - - L I W D E N D F D S
- - - - - - M Q P F R L P E F Y V P W P A R L N P H L E T A R E H S K A W A R E M G M L P G G P L G D - - D Q A V W D E A T F D A
M P A P Q Q R Q P Y R L P A F Y L P R P A R L N P D L E A A R A R S R R W A E E M G M L G S R A E P E - - G E Q V W T R E D F D R
- - - - - - M Q P F T L P D F Y M P H P A R L N P H L E G A R A H T K A W S Y E M G I L E E N P K E K - - - - P I W T E S D L D A
M A D A Q V K Q P F K L P E F Y V P W P A R L N P H L E A A R V H S K A W A Y E M G I I G V P K D G S - - A P E I W N E A K F D A
M A D A Q V K Q P F K L P E F Y V P W P A R L N P H L E A A R V H S K A W A Y E M G I I G V P K D G S - - A P E I W N E A K F D A
- - - - - V T Q P F V L P D F Y V P Y P A R L N P H L E E A R A H A R V W A D K M G M L E - - - - G S - - - - G I W D L A D L E A
- - - - - M T Q P F Q L P H F Y M P Y P A R L N P H L D E A R A H S T R W A R G M G M L E - - - - G S - - - - G I W E Q S D L D A
- - - - - M S Q P F E L P D F Y E P Y P A R L N P H L A A A R D H S K R W A L E M E M I E - - - - G S - - - - G V W D E A D F D S
- - - - - V T Q P F E L P D F Y M P Y P A R L N P N L E E A R T H T K Q W A R D M G M L E - - - - G S - - - - G I W D E H D L E S
- - - - - M A Q P F S L P D F Y V P Y P A R L N P H V E A A R T H T R A W A R A M G M L E - - - - G S - - - - G I W E E R D L E A
- - - - - M A Q P F V L P D F Y V P Y P A R L N P H V E T A R A H T R A W A R E M G M L E - - - - G S - - - - G V W E T R D L D A
- - - - M T Q Q P F Q L P H F Y L P H P A R L N P H L D E A R A H S T T W A R E M G M L E - - - - G S - - - - G V W E Q S D L E A
- - - - - M A Q P F S L P E F Y V P Y P A R L N P H L D A A R S H T R E W A R G M G M L E - - - - G S - - - - G I W E E K D L E S
- - - - - M T Q P F E L P H F Y L P H P A R L N P H V D E A R A H S T A W A R G M G M L E - - - - G S - - - - G V W E Q A D L D A
- - - - - M T Q P F H L P H F Y M P Y P A R L N P H L D E A R A H S T A W A R E M G M L E - - - - G S - - - - G V W E Q S D L D A
- - - - - M T Q P F E L P H F Y L P H P A R L N P H V E E A R A H S T G W A R E M G M L E - - - - G S - - - - G V W E Q A D L D A
- - - - - M A Q P F S L P D F Y V P Y P A R L N P H V E A A R T H T R A W A R A M G M L E - - - - G S - - - - G I W E E K D L E A
- - - - - M T Q P F Q L P D F Y M P Y P A R L N P H V Q E A R E H S T R W A R E R G M L E - - - - G S - - - - G I W E Q E D L D A
- - - - - M T Q P F E L P D F Y M P H P A R L S P H L E E A R R H S K R W A R E M G M L E - - - - G S - - - - G V W E E R D L D A
- - - - M T Q Q P F Q L P H F Y L P H P A R L N P H L D E A R A H S T T W A R E M G M L E - - - - G S - - - - G V W E Q S D L E A
- - - - - M A Q P F V L P D F Y V P Y P A R L N R H V E E A R R H S K K W A R R M G M L E - - - - G S - - - - G I W E E S D L D A
- - - - - M A Q P F V L P D F Y V P Y P A R L N P H V E E A R R H T R K W A R R M G M L E - - - - G S - - - - G I W E E S D L E A
- - - - - M T Q P F A L P H F Y L P Y P A R L N P H L E E A R A H S S V W A R E M G M L E - - - - G S - - - - G V W N Q A D L D A
- - - - - - M Q P F V L P E F Y L S Y P A R L N P H L D G A R K H S K A W A Y S M D M I D V P Q H G T - - - - V I W N E H D L D S
- - - - - M A Q P F T L P D F Y V P Y P A R L N P H L E A A R T H T R A W A R R M G M L E - - - - G S - - - - G I W E E K D L E S
- - - - - M T Q P F R L P E F Y V P Y P A R L N P H L E E A R A H S K R W A R S F G M L E - - - - G S - - - - G V W E E S D L D S
- - - - - M T Q P F Q L P D F Y V P H P A R L N P H L E E A R V H T K R W A R A L G M L E - - - - G S - - - - G V W E E S D L D S
- - - - - M A Q P F T L P D F Y V P Y P A R L N P H L E A A R V H A R A W A R S M G M L E - - - - G S - - - - G V W E Q R D L D A
- - - - - M A Q P F T L P D F Y V P Y P A R L N P H L E A A R V H A R A W A R S M G M L E - - - - G S - - - - G V W E Q R D L D A
- - - - - M A Q P F T L P D F Y V P Y P A R L N P H L E A A R V H A R A W A R S M G M L E - - - - G S - - - - G V W E Q R D L D A
R A R L M T Q Q P F E L P H F Y M P Y P A R L N P H V E E A R A H S T V W A R E M G M L E - - - - G S - - - - G I W E Q S D L D A
- - - - - M P Q P F V M P D F Y V P Y P A R L N P H L E A A R T H T R D W A R A M G M L E - - - - G S - - - - G V W E Q H D L D S
- - - - - V T Q P F Q L P E F Y M P Y P A R L S P H V Q E A R E H S T Q W A R A K G M L E - - - - G S - - - - G I W E Q K D L D A
- - - - - M T Q P F E L P H F Y L P H P A R L N P H V D E A R A H S T H W A R E M G M L E - - - - G S - - - - G V W E Q A D L D A
- - - - - - M Q A F T L P E F Y M P Y P A R I N P H M E R S R A H S A A W A R Q M G M L D A P K P G G - - - G V V W D D A E L A R
R A A P S G G Q P Y Q L P E F Y V P Y P A R I N P H L E Q A R E H S R A W A Y A F D M I D V P Q Q G K - - - - A I W D L R D F D S
- - - M T S E Q P F T L P N F Y M P Y P A R L N P N L A G A R G H T M A W A K D M G M L D S P S A G G - - - G L I W D E A E L A R
- - - - - M P K P F Q L P D F Y M P Y P A R L N P H L E F A R V Q S K G W A R G L A M I E - - - - G S - - - - G V W D E G D F D R

12
 Figure 1. Alignment of geosmin synthases.

141 151 161 171 181 191

YP_003098781 (DSM 43827)
AEK39836 (S699)
YP_003763541 (U32)
AEA03338 (CHAB 1432)
AEA03341 (CHAB 2155)
YP_716636 (ACN14a)
YP_483306 (Ccl3)
YP_001509819 (EAN1pec)
YP_004017381 (Eul1c)
YP_003265710 (DSM 14365)
BAJ30389 (KM-6054)
YP_634376 (DK 1622)
YP_001866236 (PCC 73102)
ZP_07114089 (PCC 6056)
ABU93239 (P2r)
ABU93238 (P2r)
YP_001105388 (NRRL 2338)
YP_001107098 (NRRL 2338)
YP_001106173 (NRRL 2338)
YP_001612078 (So ce 56)
ZP_01460669 (DW 4/3-1)
YP_003950745 (DW 4/3-1)
ZP_04705018 (J1074)
NP_823339 (MA-4680)
ADI05189 (BCW-1)
CCB75658 (NRRL 8057)
ZP_08240572 (XylebKG-1)
ZP_06769636 (ATCC 27064)
NP_630182 (A3(2))
ADW07414 (ATCC 33331)
ZP_04685148 (ATCC 14762)
ZP_08289513 (M045)
ZP_07309957 (Tü 4000)
YP_001828351 (NBRC 13350)
ZP_07296488 (ATCC 53653)
ZP_07293917 (ATCC 53653)
ZP_05522837 (TK 24)
ABY50951 (ATCC 27952)
ZP_06913794 (ATCC 25486)
YP_003487693 (87.22)
ZP_07276967 (AA4)
ZP_06273105 (ACTE)
ZP_07289987 (C)
ZP_05001875 (Mg1)
ZP_07976693 (SA3_actG)
ZP_07273435 (SPB78)
ZP_08453284 (Tü 6071)
ZP_06920565 (ATCC 29083)
CCA53556 (ATCC 10712)
ZP_07608000 (Tü 4113)
ZP_07307300 (DSM 40736)
YP_003335875 (DSM 43021)
YP_003382082 (DSM 17836)
YP_003509930 (DSM 44728)
YP_003116895 (DSM 44928)
Highly conserved residues are shown in red, deviations from otherwise highly conserved residues are shown in dark red. The strictly conserved motifs are shown in boxes. Added nucleotides of corrected sequences are shown in bold.
H D Y A L L C S Y T H P D A T A A D L D L V T D W Y V W V F Y F D D H F L E L Y K R N P D M A G A K A Y L D R L P L F M P V D G -
H D Y A L L C A Y T H P D A G A E E L D L I T D W Y V W V F Y F D D H F L E L Y K R T G D I D S A R G Y L D R L E L F M P A E - -
H D Y A L L C A Y T H P D A G A E E L D L I T D W Y V W V F Y F D D H F L E L Y K R T G D I D S A R G Y L D R L E L F M P A E - -
H D Y A L L C S Y T H P D T P S T E L N L V T D W Y V W V F F F D D H F L E I Y K R S Q D L I G A K E Y L D R L P A F M P I Y P -
H D Y A L L C S Y T H P D T P S T E L N L V T D W Y V W V F F F D D H F L E I Y K R S Q D L I G A K E Y L D R L P A F M P I Y P -
H D Y A L L C A Y T H P D A T A D R L N L I T D W Y V W V F Y F D D H F L E L Y K R S H D L A G A R A Y L D R L P A F M P V D - -
H D Y A L L C A Y T H P D S V S R K L D L V T D W Y V W V F Y F D D H F L E L Y K R S H D M A G A R A Y L D R L P A F M P V D - -
H D Y A L L C A Y T H P D A D E A R L D L I T D W Y V W V F Y F D D H F L E V Y K R G R D V A G A R R Y L D R L R L F M P V E - -
H D Y A L L C A Y T H P D S S A D R L N L V T D W Y V W V F Y F D D H F L E L F K R T G D M A G A R D Y L D R L R A F M P V D A G
H D Y A L L C A Y T H P D A S S A E L D L I T D W Y V W V F F F D D H F L E T F K R T R D M Q G A K Q Y L G R L P A F M P I G Q -
H D Y A L L C S Y T H P D C G P E A L D L V T D W Y T W V F F F D D H F L E T F K R T L D R E G G K A Y L D R L P A F M P M D P -
M D Y A L L C A Y T H P E A P G P E L D L V T D W Y V W V F Y F D D H F L E L Y K R P Q D Q V G A K A Y L D R L P L F M P V D P -
H D Y A L L C S Y T H P D A P G T E L D L V T D W Y V W V F F F D D H F L E I Y K R T Q D M A G A K E Y L G R L P M F M P I Y P -
H D Y A L L C S Y T H P D A P G A E L D L V T D W Y V W V F F F D D H F L E I Y K R T Q D M T G A K E Y L H R L P A F M P I H P -
H D Y A L L C A Y T H P D T P G T E L D L I T D W Y V W V F F F D D H F L E I Y K R S Q D M V G A K A Y L D R L P A F M P I F - -
H D Y A L L C A Y T H P D A P A T E L D L I T D W Y V W V F F F D D H F L E I Y K R S Q D M I G A K A Y L D R L P A F M P V Y P -
H D Y A L L C A Y T H P D A D G P M L D L I T D W Y V W V F Y F D D H F V E L Y K R N P D L A G A K E Y L D R L P A F M P V E - -
H D Y A L L C A Y T H P D A T A H E L G L V T D W Y V W V F Y F D D H F L E Y Y K R T R D L T G A R E Y L A G L A A F M P A E L -
H D Y A L L C A Y A H P D A S A P A L E L I T G W Y V W A F F F D D H F L A R Y K R T G D V D G A R A H L L G L A E L M P V G P -
H D Y A L L C A Y T H P D A P A P E L N L I T D W Y V W V F F F D D H F L E I Y K R T K D M R G A K A Y L D R L P L F M P V V P -
M D Y A L L C A Y T H P E A P S L E L D L V T D W Y V W V F Y F D D H F L D V Y K R T Q D Q V G A R E Y L D R L P A F M P V D L -
M D Y A L L C A Y T H P E A P S L E L D L V T D W Y V W V F Y F D D H F L D V Y K R T Q D Q V G A R E Y L D R L P A F M P V D L -
H D Y A L L C A Y T H P D C D G P A L S L V T D W Y V W V F F F D D H F L E L F K R T Q D R E G A K A Y L E R L P A F M P M D L -
H D Y G L L C A Y T H P D C D G P A L S L I T D W Y V W V F F F D D H F L E T F K R T Q D R E G G K A Y L D R L P L F M P L D L -
H D Y A L L C A Y T H P D A P A E V L A T V T D W Y V W V F F F D D H F L E S F K R S R D M A G A K A Y L D R L R A F M P V L P A
H D Y A L L C A Y T H P D T T G P K L S L V T D W Y V W V F F F D D H F L E T F K R S Q D R A G G K A Y L D R L P E F M P M D L -
H D Y A L L C A Y T H P D C S A E A L S L V T D W Y V W V F F F D D H F L E L F K R T P D R E G G K R Y L D R L P A F M P M G R -
H D Y A L L C A Y T H P E C D A E A L N L V T D W Y T W V F F F D D H F L E Q F K R S L D R A G G K A Y L D R L P A F M P L D P -
H D Y G L L C A Y T H P D C D G P A L S L I T D W Y V W V F F F D D H F L E K Y K R S Q D R L A G K A H L D R L P L F M P L D D -
H D Y A L L C A Y T H P D C S S E A L S L V T D W Y V W V F F F D D H F L E L F K R T P D R E G G R K Y L D R L P A F M P M E R -
H D Y G L L C A Y T H P D C D G P A L C L I T D W Y V W V F F F D D H F L E R F K R T Q D R D G G K A H L D R L P L F M P A D P -
H D Y G L L C A Y T H P D C D G P A L S L I T D W Y V W V F F F D D H F L E L F K R T Q D R A A G K A H L D R L P L F M P A D P -
H D Y G L L C A Y T H P D C D G P A L S L V T D W Y V W V F F F D D H F L E K F K R S Q D R T A G K A H L D R L P L F M P L D P -
H D Y A L L C A Y T H P D C S A E A L S L V T D W Y V W V F F F D D H F L E L F K R T P D R E G G K R Y L D R L P A F M P M G R -
H D Y A L L C A Y T H P D C S A A E L A L I T D W Y V W V F F F D D H F L E R F K R S Q D R A G G K A Y L D R L P L F M P M D P A
H D Y A L L C A Y T H P D A S G P A L S L V T D W Y V W V F F F D D H F L E E F K Y T N D R E G A K A Y L D R L P L F M P T D P -
H D Y G L L C A Y T H P D C D G P A L S L I T D W Y V W V F F F D D H F L E K Y K R S Q D R L A G K A H L D R L P L F M P L D D -
H D Y A L L C A Y T H P D C D A D A L G L V T D W Y V W V F F F D D H F L E V F K R S Q D L A G G K A Y L D R L P A F M P M D L -
H D Y A L L C A Y T H P D C D A Q A L G L V T D W Y V W V F F F D D H F L E V F K R S Q D L A G A K A Y L G R L P A F M P M D L -
H D Y G L L C A Y T H P D C D G P A L S L I T D W Y V W V F F F D D H F L E L Y K R S Q D R P G G K A H L D R L P L F M P L D L -
H D Y A L L C S Y T H P D A A P R E L D L V T D W Y V W V F Y F D D H F L E L F K R T G D I E H A R A Y L D R I A R F M P A E - -
H D Y A L L C A Y T H P D C S D E A L S L V T D W Y V W V F F F D D H F L E L F K R T P D R E G G K R Y L D R L P A F M P M E R -
H D Y A L L C S Y T H P D C D A E A L S L V T D W Y V W V F F F D D H F L E T F K R S R D R A G A K A Y L D R L P A F M P M D L -
H D Y A L L C A Y T H P D C D S E A L S L V T D W Y V W V F F F D D H F L E M Y K R S Q D R A G A K A Y L D R L A A F M P M D L -
H D Y A L L C A Y T H P D C D E E A L N L V T D W Y V W V F F F D D H F L E L F K R G Q D R E G G K A Y L D R L P A F M P A D L -
H D Y A L L C A Y T H P D C D E E A L N L V T D W Y V W V F F F D D H F L E L F K R G Q D R E G G K A Y L D R L P A F M P A D L -
H D Y A L L C A Y T H P D C D E E A L N L V T D W Y V W V F F F D D H F L E L F K R G Q D R E G G K A Y L D R L P A F M P A D L -
H D Y G L L C A Y T H P D C D G P A L S L I T D W Y V W V F F F D D H F L E T F K R T Q D R A G G K A Y L D R L P L F M P M D L -
H D Y A L L C S Y T H P D C D E E A L N L V T D W Y T W V F F F D D H F L E I Y K R P Q D R S G G K A Y L D R L P L F M P A D P -
H D Y A L L C A Y T H P D C S G A E L S L I T D W Y V W V F F F D D H F L E T F K R S Q D R E A G K T Y L D R L P A F M P M D P -
H D Y G L L C A Y T H P D C D A P A L S L I T D W Y V W V F F F D D H F L E M Y K R S Q D R V A G K A H L D R L P L F M P L D L -
M D Y A L M C A Y T H P D C D G P T L D L I T D W Y V W V F F F D D H F L E Q F K Y S R D L L G A K A Y L D H L E L F M T A D - -
H D Y A L L C A Y T H P D A G A A A L D L V T D W Y V W V F Y F D D H F L E L Y K K T Q D T A G A K A Y L N R L A A F M P V D - -
H D Y G L L C A Y T H P D C D Q A M L D L I T D W Y V W V F F F D D H F L E Q F K R T R D T V G A K E Y L D R L H L F M P V S - -
H D Y A L L C S Y T H P D C D A E E L A L V T D W Y V W V F F F D D H F W E I Y K R P R D M V G A Q A Y L D R L P A F M P I G D -

13
 Figure 1. Alignment of geosmin synthases.

201 211 221 231 241 251 261

YP_003098781 (DSM 43827)
AEK39836 (S699)
YP_003763541 (U32)
AEA03338 (CHAB 1432)
AEA03341 (CHAB 2155)
YP_716636 (ACN14a)
YP_483306 (Ccl3)
YP_001509819 (EAN1pec)
YP_004017381 (Eul1c)
YP_003265710 (DSM 14365)
BAJ30389 (KM-6054)
YP_634376 (DK 1622)
YP_001866236 (PCC 73102)
ZP_07114089 (PCC 6056)
ABU93239 (P2r)
ABU93238 (P2r)
YP_001105388 (NRRL 2338)
YP_001107098 (NRRL 2338)
YP_001106173 (NRRL 2338)
YP_001612078 (So ce 56)
ZP_01460669 (DW 4/3-1)
YP_003950745 (DW 4/3-1)
ZP_04705018 (J1074)
NP_823339 (MA-4680)
ADI05189 (BCW-1)
CCB75658 (NRRL 8057)
ZP_08240572 (XylebKG-1)
ZP_06769636 (ATCC 27064)
NP_630182 (A3(2))
ADW07414 (ATCC 33331)
ZP_04685148 (ATCC 14762)
ZP_08289513 (M045)
ZP_07309957 (Tü 4000)
YP_001828351 (NBRC 13350)
ZP_07296488 (ATCC 53653)
ZP_07293917 (ATCC 53653)
ZP_05522837 (TK 24)
ABY50951 (ATCC 27952)
ZP_06913794 (ATCC 25486)
YP_003487693 (87.22)
ZP_07276967 (AA4)
ZP_06273105 (ACTE)
ZP_07289987 (C)
ZP_05001875 (Mg1)
ZP_07976693 (SA3_actG)
ZP_07273435 (SPB78)
ZP_08453284 (Tü 6071)
ZP_06920565 (ATCC 29083)
CCA53556 (ATCC 10712)
ZP_07608000 (Tü 4113)
ZP_07307300 (DSM 40736)
YP_003335875 (DSM 43021)
YP_003382082 (DSM 17836)
YP_003509930 (DSM 44728)
YP_003116895 (DSM 44928)
Highly conserved residues are shown in red, deviations from otherwise highly conserved residues are shown in dark red. The strictly conserved motifs are shown in boxes. Added nucleotides of corrected sequences are shown in bold.
- - - - P I T - - - E E P T N P V E K G L A D L W A R T T P V H T A D W R R R F A D N T K H L L D E S L W E L A N I S E G R L S N
- - G E I T A - - - D P - E N P V E R G L T D L W N R T V P H R S A G W R R R F A D S T K A L L D E S L W E L A N I N E G R L A N
- - G E I T A - - - D P - E N P V E R G L T D L W N R T V P H R S A G W R R R F A D S T K A L L D E S L W E L A N I N E G R L A N
- - Q D N L P - - - F P - T N P V E R G L A D L W S R T A F T K S V E W R Q R F F E S T K N L L D E S M W E L A N I N Q N R I A N
- - Q D N L P - - - F P - T N P V E R G L A D L W S R T A F T K P V E W R Q R F F E S T K N L L D E S M W E L A N I N Q N R I A N
- - G E I T E - - - E P - S N P V E R G L A D L W T R T V P A R S A D W R A R F A V S T R N L L D E S L W E L E N I N A A R L S N
- - - G E I T - - - E T P T N P V E R G L A D L W T R T V P E R S A D W R R R F A V S T K N L L D E S L W E L A N I N A G R L A N
- - G A V T A - - - E P - A N P V E R G L A D L W S R T V P D R T P A W R R R F A T S T R H L L D E S L W E L A N I D E N R L A N
A V A G E A P T G Q D A P T N P V E R G L A D L W A R T V P D R S A A W R Q R F V V S T R N L L D E S L W E L A N I N A N R V A N
- - S K G A L - - - E P - S N A V E R G L V N L W A R T I P T A S A D W Q R R F K L H N E H L L E E S L W E L T N I R A E R I S N
- - E A G Y P - - - E P - T N P V E A G L A D L W R R T V P H M S A D W R A R F A E S T R N L L N E S L W E L S N I N E G R I S N
- - - A A T P - - - P P P T N P V E A G L L D L W N R T V P S R S M A W R R R F F E S T K H L L D E S S W E L S N I S D R R V S N
- - - T E T P - - - P V P T N P V E C G L A D L W S R T A F T K S V D W R L R F F E S T K N L L E E S L W E L A N I N Q D R V A N
- - - T D T P - - - S L P T N P V E R G L A D L W S R T A F T K S V D W R L R F F E S T K N L L E E S L W E L A N I N Q N R V A N
- - A D E T P - - - P V P T N P V E R G L L N L W A R T V F T K S V E W R R R F F A S T K H L L D E S M W E L A N I N Q D R I A N
- - - T E A P - - - P V P T N P V E R G L S N L W F R T A F T K S V E W R Q R F F E S T K Y L L E E S M W E L A N I N Q N R I A N
- - G P I T A - - - E P - T N P V E R G L A D L W Q R T V P A R T A D W R R R Y A E N T K H L L D E S L W E L S N I S R N R L S N
- - T A E Q P - - - - T A K N P V E W G L V D L W A R S V P I M S A D W L R R F S E S T R N L L E D C V W E L T N I T H G Q V P N
- - S D A A P - - - - A A T G P V E R G L A D L W V R T A P E V P A R W L V R F A A S T R E L L E N R L R E L T G T S R C G V P N
- - - T G T P - - - P E P T N V V E R A L A D L W A R T V P L T S V H W R E R F L E A T K N L M L E C M W E L A N I Q E N R V A N
- - - S A A P - - - P T P T N P V E R G L A D L W A R T V P T K S E A W R R R F F E S T K S L L E E S N W E L N N I S E R R V S N
- - - S A A P - - - P T P T N P V E R G L A D L W A R T V P T K S E A W R R R F F E S T K S L L E E S N W E L N N I S E R R V S N
- - A D G F P - - - E P - E N P V E A G L A D L W A R T V P A M S E D W R R R F A L S T E N L L N E S M W E L S N I N A G R V S N
- - S A P V P - - - E P - E N P V E A G L A D L W A R T V P A M S A D W R K R F A V S T E H L L N E S L W E L S N I N E G R I A N
A R A A R A A - - - G E P A N A V E R G L A D L W A R T T P A M S G G W R R R F A E A T R H L L E E S M W E L S N I S E G R I A N
- - A A G F P - - - E P - T N P V E A G L A D L W A R T V P S M S L A W R T R F A E S T A N L L N E S L W E L S N I N A D R V P N
- - G A P T P - - - E P - E N P V E A G L A D L W A R T V P S M S D A W R A R F A E A T E A L L N E S L W E L A N I H E G R V A N
- - A E A A P - - - - P A T N P V E A G L A D L W T R T V P A M S P A W R A R F T E S T R N L L N E S L W E L S N I H E G R V A N
- - A A G M P - - - E P - R N P V E A G L A D L W T R T V P A M S A D W R R R F A V A T E H L L N E S M W E L S N I N E G R V A N
- - G A A T P - - - E P - A N P V E A G L A D L W T R T V P A M S D A W R A R F A E S T A N L L N E S L W E L S N I N E G R I A N
- - A A P V P - - - - A P E N P V E A G L A D L W A R T V P A M S A D W R R R F A L S T E H L L N E S L W E L S N I N E G R I A N
- - A T P V P - - - E P - Q N P V E A G L A D L W A R T V P A M S A D W R R R F A V A T E H L L N E S L W E L S N I N E G R I S N
- - A T P V P - - - E P - E N P V E A G L A D L W A R T V P A M S A D W R R R F A V A T E H L L N E S M W E L S N I N E G R I A N
- - G A P T P - - - E P - E N P V E A G L A D L W A R T V P S M S D A W R A R F A E A T E A L L N E S L W E L A N I H E G R V A N
- - A A G M P - - - E P - E N P V E A G L K D L W L R T V P A M S P D W R A R F R E S T E N L L N E S L W E L S N I N A G R T P N
- - A T A V P - - - E P - V N P V E A G L A D L W T R T V P S M S A G W R A R F A E S T E N L L N E S L W E L S N I N A H R I P N
- - A A G M P - - - E P - R N P V E A G L A D L W T R T V P A M S A D W R R R F A V A T E H L L N E S M W E L S N I N E G R V A N
- - S R G T P - - - E P - R N P V E A G L A D L W Q R T V P S M S P A W R T R F A E A T E H L L N E S M W E L T N I D A G R V A N
- - S Q G T P - - - E P - T N P V E A G L A D L W E R T V P S M S P A W R A R F A E S T K N L L D E S M W E L A N I D A G R V A N
- - S T P V P - - - E P - R N P V E A G L A D L W A R T V P S M S M D W R R R F A V A T E H L L N E S M W E L S N I N E G R I A N
- - G E I T - - - - E T P E N P V E R G L T D L W N R T V P H R S A G W R H R F R E S T R N L L D E S L W E L A N I N E S R I A N
- - G A P T P - - - E P - T N P V E A G L A D L W A R T V P A M S D A W R A R F A E A T E N L L N E S L W E L A N I N E G R I A N
- - A E G F P - - - - E A V N P V E A G L A D L W A R T V P A M S A D W R E R F S L S T K N L L D E S M W E L A N I G I G R V A N
- - A D G F P - - - E P - A G P V E A G L A D L W E R T V P A M S P H W R E R F A E S T R N L L N E S M W E L A N I N I G R V A N
- - A D G F P - - - E P - E N P V E A G L A D L W R R T V P A M S A D W R E R F S T S T R N L L N E S L W E L S N I N A G R I S N
- - A D G F P - - - E P - E N P V E A G L A D L W R R T V P A M S A D W R E R F S T S T R N L L N E S L W E L S N I N A G R I S N
- - A D G F P - - - E P - E N P V E A G L A D L W R R T V P A M S A D W R E R F S T S T R N L L N E S L W E L S N I N A G R I S N
- - S T P M P - - - E P - E N P V E A G L A D L W T R T V P S M S V D W R R R F A V A T E H L L N E S L W E L S N I N E G R I A N
- - A A G M P - - - E P - T N P V E A G L A D L W L R T V P S M S E G W R V R F A E A T E H L L Y E S L W E L D N I N D G R V A N
- - A A G T P - - - E P - T N P V E A G L A D L W A R T V P S M S S D W R A R F R E S T E N L L N E S L W E L S N I N I H R V P N
- - S T S V P - - - E P - E N P V E A G L A D L W A R T V P K M S Q D W R R R F A V A T E H L L N E S M W E L S N I N E G R I A N
- - - G E T P - - - P E P A N P A E A G L K D L W E R T V P A M S H G W R Q R F I T S T H N L M V E S M W E L D N I D R G R I A N
- - G A I T - - - - E T P E N P V E R G L A D L W A R T V P S M S E G W R R R F A A T T E S L L A E S L W E L S N I S A G R L P N
- - G Q E L P - - - E P - T N P V E K G L A D L W Q R T A P H R S E D W K Q R F S V S T R N L L M E S L W E L S N I T T G R V A N
- - L G A M P - - - E P - T N A V E A G L A D L W L R T V P S K S E P W R R R F A E S N R H L L A E S L W E L A N I T E D R V S D

14
 Figure 1. Alignment of geosmin synthases.

271 281 291 301 311 321

YP_003098781 (DSM 43827)
AEK39836 (S699)
YP_003763541 (U32)
AEA03338 (CHAB 1432)
AEA03341 (CHAB 2155)
YP_716636 (ACN14a)
YP_483306 (Ccl3)
YP_001509819 (EAN1pec)
YP_004017381 (Eul1c)
YP_003265710 (DSM 14365)
BAJ30389 (KM-6054)
YP_634376 (DK 1622)
YP_001866236 (PCC 73102)
ZP_07114089 (PCC 6056)
ABU93239 (P2r)
ABU93238 (P2r)
YP_001105388 (NRRL 2338)
YP_001107098 (NRRL 2338)
YP_001106173 (NRRL 2338)
YP_001612078 (So ce 56)
ZP_01460669 (DW 4/3-1)
YP_003950745 (DW 4/3-1)
ZP_04705018 (J1074)
NP_823339 (MA-4680)
ADI05189 (BCW-1)
CCB75658 (NRRL 8057)
ZP_08240572 (XylebKG-1)
ZP_06769636 (ATCC 27064)
NP_630182 (A3(2))
ADW07414 (ATCC 33331)
ZP_04685148 (ATCC 14762)
ZP_08289513 (M045)
ZP_07309957 (Tü 4000)
YP_001828351 (NBRC 13350)
ZP_07296488 (ATCC 53653)
ZP_07293917 (ATCC 53653)
ZP_05522837 (TK 24)
ABY50951 (ATCC 27952)
ZP_06913794 (ATCC 25486)
YP_003487693 (87.22)
ZP_07276967 (AA4)
ZP_06273105 (ACTE)
ZP_07289987 (C)
ZP_05001875 (Mg1)
ZP_07976693 (SA3_actG)
ZP_07273435 (SPB78)
ZP_08453284 (Tü 6071)
ZP_06920565 (ATCC 29083)
CCA53556 (ATCC 10712)
ZP_07608000 (Tü 4113)
ZP_07307300 (DSM 40736)
YP_003335875 (DSM 43021)
YP_003382082 (DSM 17836)
YP_003509930 (DSM 44728)
YP_003116895 (DSM 44928)
Highly conserved residues are shown in red, deviations from otherwise highly conserved residues are shown in dark red. The strictly conserved motifs are shown in boxes. Added nucleotides of corrected sequences are shown in bold.
P I E Y V E M R R K V G G A P W S A N L V E H V T G M E V P A A I A L S R P L G V L R D T F S D A V H L R N D L F S Y E R E V Q D
P I E Y V E M R R K V G G A P W S A N L V E H S V H A E V P G A I A A S R P M E V L R D C F A D S V H L R N D L F S Y Q R E V Q D
P I E Y V E M R R K V G G A P W S A N L V E H S V H A E V P G A I A A S R P M E V L R D C F A D S V H L R N D L F S Y Q R E V Q D
P I E Y I E M R R K V G G A P W S A D L V E H A A F V E V P A K I A A T R P M R V L K D T F A D G V H L R N D L F S Y Q R E V E E
P I E Y I E M R R K V G G A P W S A D L V E H A A F V E V P A K I A A T R P M R V L K D T F A D G V H L R N D L F S Y Q R E V E E
P I E Y I E M R R K V G G A P W S A N L V E H A A D A E V P A R V A A T R P L Q V L R D T F A D A V H L R N D L F S Y E R E V T E
P I E Y V E M R R K V G G A P W S A N L V E H A A D A E V P A Q V A A T R P L Q V L R D T F A D A V H L R N D L F S Y Q R E V E E
P V E Y I E M R R K V G G A P W S A N L V E H A A D A E V P D A I A A T R P A Q V L R D T F S D A I H L R N D L F S Y Q R E V Q E
P I E Y I E M R R K V G G A P W S A N L V E H A A D A E V P A A I A G L R P M R V L R D T F A D A I H L R N D L F S Y Q R E V E L
P I E Y I E M R R K V G G A P W S A N L I E H A H G I E V P A R V V D A R P M K V L T A T F A D A V H L R N D L F S Y E R E V N E
P V E Y I E M R R K V G G A P W S A G L I E Y A A N A E V P E S V A H S R P L R V L R D A F S D S V H L R N D L F S Y E R E I G E
P I E Y I E M R R K V G G A P W S A N L V E H A V F A E V P D R V A A S R P M R V L K D T F S D A V H L R N D L F S Y E R E I L E
P I E Y I E M R R K V G G A P W S A D L V E H A V F I E I P A D I A S T R P M R V L K D T F A D G V H L R N D L F S Y Q R E V E D
P I E Y I E M R R K V G G A P W S A D L V E H A A F V E V P A Q I A A T R P M R V L K D T F A D G V H L R N D I F S Y Q R E V E D
P I E Y I E M R R K V G G A P W S A D L V E H A C F V E V P A K I A A T R P M R V L K D T F A D G V H L R N D L F S Y Q R E V E D
P I E Y I E M R R K V G G A P W S A D L V E H A V F I E V P A K I A A T R P M R V L K D T F S D G V H L R N D L F S Y Q R E V E D
P I E Y I E M R R K V G G A P W S A N L V E H A V D S E V P A A I A S A R P M Q V L R D T F S D A V H L R N D L F S Y Q R E V Q D
P I D Y V E M R R R V G G A P W S A D L V E L A A R V E V P A Q I A R T R P M S V L K D T F A D A V H L R N D I F S Y Q R E T E E
P V D H I A M R R E A G G A S W S A A L V E Y A A G S E V P D V V A R S R P M R V L R D S F C D G V H L R N D I F S Y P R E T S E
P V E Y I E M R R K V G G A P W S A G L V E I A V G A E V P A A V A A T R P L E V L R D T F S D G V H L R N D L F S Y Q R E V E V
P I E Y I E M R R K V G G A P W S A D L V E H A V F A E I P A R I A A S R P M T V L K D T F S D G V H L R N D L F S Y Q R E I Q E
P I E Y I E M R R K V G G A P W S A D L V E H A V F A E I P A R I A A S R P M T V L K D T F S D G V H L R N D L F S Y Q R E I Q E
P L E Y I E M R R K V G G A P W S A G L I E Y A A Q A E V P E A V A Y S R P L R V L T D A F S D G V H L R N D L F S Y Q R E V E E
P V E Y I E M R R K V G G A P W S A G L V E Y A T - A E V P A A V A G S R P L R V L M E T F S D G V H L R N D L F S Y Q R E V E E
P L E Y V E M R R K V G G A P W S A G L V E Y A A G A E V P E A V A A S R P L L V L R D A F A D A V H L R N D L F S Y Q R E V E D
P V E Y I E M R R K V G G A P W S A G L V E F A V G A E V P D V I A D S R P M R V L R D S F A D G V H L R N D L F S Y Q R E T E E
P V E Y I E M R R K V G G A P W S A G L V E Y A A G A E V P A S V A D A R P L R V L R D A F S D A V H L R N D L F S Y Q R E V E D
P V E Y I E M R R K V G G A P W S A Q L V E Y A V G A E V P A A V A G S R P L R V L T D T F A D A V H L R N D L F S Y Q R E V E D
P V E Y I E M R R K V G G A P W S A G L V E Y A T - A E V P A A V A G T R P L R V L M E T F S D A V H L R N D L F S Y Q R E V E D
P V E Y I E M R R K V G G A P W S A G L V E Y A A N A E V P A S L A D A R P L R V L R D A F S D G V H L R N D L F S Y Q R E V E D
P V E Y I E M R R K V G G A P W S A G L V E Y A T - A E V P A A V A G T R P L R V L M E T F S D A V H L R N D L F S Y Q R E V E D
P V E Y I E M R R K V G G A P W S A G L V E Y A T - A E V P A R V A E S R P L R V L M E T F S D G V H L R N D L F S Y Q R E V E D
P V E Y I E M R R K V G G A P W S A G L V E Y A T - A E V P A A V A G T R P L R V L M E T F S D A V H L R N D L F S Y Q R E V E D
P V E Y I E M R R K V G G A P W S A G L V E Y A A G A E V P A S V A D A R P L R V L R D A F S D A V H L R N D L F S Y Q R E V E D
P V E Y I E M R R K V G G A P W S A G L V E H A V G A E V P A V I A E S R P L R V L R D A F A D A V H L R N D L F S Y Q R E I E E
P V E Y I E M R R K V G G A P W S A G L V E Y A I G A E V P A P V A A S R P L Q V L R D S F S D A V H L R N D L F S Y Q R E V E E
P V E Y I E M R R K V G G A P W S A G L V E Y A T - A E V P A A V A G T R P L R V L M E T F S D A V H L R N D L F S Y Q R E V E D
P V E Y I E M R R K V G G A P W S A G L V E Y A A Q A E V P E S V A G A R P L R V L R D S F S D A V H L R N D L F S Y Q R E V E D
P L E Y I E M R R K V G G A P W S A G L V E Y A A G A E V P E Q V A M T R P L R V L R D S F S D A V H L R N D L F S Y Q R E V E D
P V E Y I E M R R K V G G A P W S A G L V E Y A T - A E V P E S V A D T R P L R V L M E T F S D A V H L R N D L F S Y Q R E V E E
P I E Y V A M R R K V G G A P W S A N L I E H S L N A E V P D E I A A K R P M E V L R D C F A D A V H L R N D L F S Y Q R E V E D
P V E Y I E M R R K V G G A P W S A G L V E Y A A H A E V P A S L A D A R P L R V L R D A F S D G V H L R N D L F S Y Q R E V E D
P L E Y I E M R R K V G G A P W S A G L V E Y V A - A E V P A R V A H S R P L G V L R D A F S D A V H I R N D L F S Y Q R E V E D
P L E Y I E M R R K V G G A P W S A G L V E Y V S - A E V P A R V A H T R P L A V L R D A F S D A V H I R N D L F S Y E R E V A D
P V E Y I E M R R K V G G A P W S A G L V E Y A A G A E L P A R V A G T R P L R V L T D A F S D A V H L R N D L F S Y Q R E V E D
P V E Y I E M R R K V G G A P W S A G L V E Y A A G A E L P A R I A G T R P L R V L T D A F S D A V H L R N D L F S Y Q R E V E D
P V E Y I E M R R K V G G A P W S A G L V E Y A A G A E L P A R V A G T R P L R V L T D A F S D A V H L R N D L F S Y Q R E V E D
P V E Y I E M R R K V G G A P W S A G L V E Y A T - A E V P A A V A G S R P L R V L M E T F S D A V H L R N D L F S Y Q R E V E D
P V E Y I E M R R K V G G A P W S A G L V E Y A A G A E V P A Q V A Y S R P L R V L R D A F S D A V H L R N D L F S Y Q R E V E D
P V E Y I E M R R K V G G A P W S A G L V E H A V G A E V P A M I A G S R P M R V L R D A F A D A V H L R N D L F S Y Q R E I E D
P V E Y I E M R R K V G G A P W S A G L V E Y A T - A E V P M S V A R S R P L R V L M E T F S D A V H L R N D L F S Y Q R E V E D
P I E Y V Q M R R R V G G A P W S A N L V E Y A V G A E I P D G L A G T R P M R V L S D T F S D A V H L R N D L F S Y Q R E V Q E
P I E Y V E M R R K V G G A P W S A N L V E Y A V Q A E V P E E V A H A R P L K V L S D T F S D A V H L R N D L F S Y Q R E V E A
P I E Y V E M R R K V G G A P W S A N L I E H A V N A E V P A R L A P T R P L R V L R D T F S D A V H L R N D L F S Y Q R E V Q D
P V D Y I E M R R K V G G A P W S A H L V E H A N G V E V P D R V A A S R P L R V L K D T F A D A V H L R N D L F S Y Q R E V E Q

15
 Figure 1. Alignment of geosmin synthases.

331 341 351 361 371 381 391

YP_003098781 (DSM 43827)
AEK39836 (S699)
YP_003763541 (U32)
AEA03338 (CHAB 1432)
AEA03341 (CHAB 2155)
YP_716636 (ACN14a)
YP_483306 (Ccl3)
YP_001509819 (EAN1pec)
YP_004017381 (Eul1c)
YP_003265710 (DSM 14365)
BAJ30389 (KM-6054)
YP_634376 (DK 1622)
YP_001866236 (PCC 73102)
ZP_07114089 (PCC 6056)
ABU93239 (P2r)
ABU93238 (P2r)
YP_001105388 (NRRL 2338)
YP_001107098 (NRRL 2338)
YP_001106173 (NRRL 2338)
YP_001612078 (So ce 56)
ZP_01460669 (DW 4/3-1)
YP_003950745 (DW 4/3-1)
ZP_04705018 (J1074)
NP_823339 (MA-4680)
ADI05189 (BCW-1)
CCB75658 (NRRL 8057)
ZP_08240572 (XylebKG-1)
ZP_06769636 (ATCC 27064)
NP_630182 (A3(2))
ADW07414 (ATCC 33331)
ZP_04685148 (ATCC 14762)
ZP_08289513 (M045)
ZP_07309957 (Tü 4000)
YP_001828351 (NBRC 13350)
ZP_07296488 (ATCC 53653)
ZP_07293917 (ATCC 53653)
ZP_05522837 (TK 24)
ABY50951 (ATCC 27952)
ZP_06913794 (ATCC 25486)
YP_003487693 (87.22)
ZP_07276967 (AA4)
ZP_06273105 (ACTE)
ZP_07289987 (C)
ZP_05001875 (Mg1)
ZP_07976693 (SA3_actG)
ZP_07273435 (SPB78)
ZP_08453284 (Tü 6071)
ZP_06920565 (ATCC 29083)
CCA53556 (ATCC 10712)
ZP_07608000 (Tü 4113)
ZP_07307300 (DSM 40736)
YP_003335875 (DSM 43021)
YP_003382082 (DSM 17836)
YP_003509930 (DSM 44728)
YP_003116895 (DSM 44928)
Highly conserved residues are shown in red, deviations from otherwise highly conserved residues are shown in dark red. The strictly conserved motifs are shown in boxes. Added nucleotides of corrected sequences are shown in bold.
E G E L S N G V L V L E R F L D V D T Q S A A E A V N D L L T S R L H Q F E H T A L V E V P P L L D E H G V D P L G R L A V L A Y
E G E L S N G V L V F E K F L G L G T Q E A A D A V N D L I T S R L H Q F E H T A L T E V P A L L D D H C V D P A A R A A T F A Y
E G E L S N G V L V F E K F L G L G T Q E A A D A V N D L I T S R L H Q F E H T A L T E V P A L L D D H C V D P A A R A A T F A Y
E G E N S N C V L V V E R F L N V S T Q E A A N L T N E L L N S R L Y Q F D N T A V T E L P S L F E E Y G V D P V E R V N V L L Y
E G E N S N C V L V V E R F L N V S T Q E A A N L T N E L L N S R L Y Q F D N T A V T E L P S L F E E Y G V D P V E R V N V L L Y
E G E L S N G V L V V E R F L D I D T Q A A A D T V N D L L T S R L H Q F E H T A A T E L P A V L D E H A I D P A G R L A A L A Y
E G E L S N G V L V I E R F L G C G T Q E A A D T V N D L L T S R L H Q F E H T A V T E L P A V L E E H G V D P G S R L E V L A Y
E G E L S N G V L V L E R F L D C P T Q Q A A D A V N D L L T S R L H Q F E H T A L T E L P P V L D E H G V T P T A R R D V L A Y
E G E L S N G V L V I E R F L D C D T Q S A A E L V N D L L T S R L H Q F E H T A I S E V P P V L E E N G I D P L A R A A V V A Y
E G E L A N C V L V T E T F L G C D T Q R A A D L T N D L L T S R L Q Q F E H T A A T E V P L M C E E Y A L E P A E R A G V L A Y
E G E L S N G V L V L E T F L G C S T Q Q A A D A V N D L L T S R L Q Q F E N T A L T E L A P L C A E H A L D P E E A G R V L A Y
E G E L S N G V L V M E K F L N I S P P S A A H L V N E V L T S R L Q Q F E N T V L T E L P S L F V E F G L N P V E Q A Q V L T Y
E G E N A N C V L V L E R F L N V S T Q E A A N L T N E L L T S R L Y Q F D N T A V T E L P P L F E E Y G L D P V A R V N V L L Y
E G E N A N C I L V L E R F L N V S T Q E A A N L T N E L L T S R L Y Q F D N T A V T E L P P L F E E H G L D P A A R V S V L L Y
E G E N S N C V L V V E K F L N I S T Q D A A N L T N D L L N S R L Y Q F E N T A V T E L P F L F E E H G V D P V E R V N V L L Y
E G E N S N C V L V V E K F L N V N P Q D A A N L T N D L L T S R L H Q F E N T A I T E L P L L F A E Y G I D P V E Q A N V L L Y
E G E L S N S V L V F E K F L D C S T Q D A A D T V N D L L T S R L H Q F E H T A L T E V P A L L D E N G V D P Q G R L A V L G Y
E G E L N N G V L V F E R F L D C G P Q E A A D T T N E L L T S R L Q Q F E N T A L T E V P P L C E E Y G L D P A E R A A V L T Y
E G E L G N G V L V V E R F F D T D P Q E A A D T V N D L L T S R L H Q F E N V T L T E L P A M F E E H G L S P V E R A D V L D Y
E G E N A N C V L V L E R F L G V P T Q K A A D L T N E I L T S R L Q Q F E N T A L V E V P A L C V E Y G L T P A Q A L D V A R Y
E G E L A N C V L V F E K F L N V D A Q R A A N L V N E V L T S R L Q Q F E N T A L T E L P S L F E E N A L N P V E R A H V L T Y
E G E L A N C V L V F E K F L N V D A Q R A A N L V N E V L T S R L Q Q F E N T A L T E L P S L F E E N A L N P V E R A H V L T Y
E G E L S N G V L V L E R F L E C T T Q E A A E A V N D L L T S R L Q Q F E N T A L T E L G P L F A D K G L H P A E S A S V L A Y
E G E L S N G V L V L E T F F G C T T Q E A A E T V N D I L T S R L H Q F E H T A L T E V P A L A L E K G L T P P E V A A V A A Y
E G E L S N G V L V F E R F L G C D T Q A A A E A V N D L I T S R L Q Q F E H T V F T E L P P L F V E S G L D P R S C A D V L T Y
E G E F S N G V L V L E K F L H C T T Q E A A D S V N D L L T S R L Q Q F E N T A L T E V P P L L L E K G I D P K S C A D V M A Y
E G E N S N G V L V L E K F L G C S T Q E A A E A V N D L L T S R L Q Q F E N T A L T E L G P L C A E K A L D P A Q T A A V L A Y
E G E L S N G V L V L E T F L G C T T Q E A A E T V N D L L T S R L Q Q F E N T A L T E V P R L C A E L D L P P S D C A A I A L Y
E G E L S N G V L V L E T F F G C T T Q E A A D L V N D V L T S R L H Q F E H T A F T E V P A V A L E K G L T P L E V A A V G A Y
E G E N S N G V L V L E R F L D C S T Q E A A D A V N D L L T S R L H Q F E N T A L T E L P P L C A E K G L G P D E T A A V L A Y
E G E L S N G V L V L E T F F G C S T Q E A A D T V N D V L T S R L H Q F E H T A F T E V P A V A L E K G L T P D Q V A A V A A Y
E G E L S N G V L V L E T F F G C T T Q E A A E A V N D I L T S R L H Q F E H T A L T E V P A L A L E H G L A P D E V A A I A A Y
E G E L S N G V L V L E T F F G C T T Q E A A D I V N D V L T S R L H Q F E H T A F T E V P A V A L E K G L D P A G A A A V A A Y
E G E N S N G V L V L E K F L G C S T Q E A A E A V N D L L T S R L Q Q F E N T A L T E L G P L C A E K V L D P A Q T A A V L A Y
E G E R S N G V L V L E T F L G C T T Q Q A A D A V N D L L T S R L Q Q F E N T T F T E L A P L F A E H G L D A V A C A D V M A Y
E G E N S N G V L V L E T F L G C A T Q E A A D A V N D L L T S R L Q Q F E N T V F T E L P P L F A E T G L S P D Q C A A V L A Y
E G E L S N G V L V L E T F F G C T T Q E A A E L V N D V L T S R L H Q F E H T A F T E V P A V A L E N G L T P P E V A A V G A Y
E G E N S N G V L V L E R F L G C G T Q E A A E V V N D L L T S R V Q Q F E N T A L T E V P A L C V Q K G L A P A E C A A I A A Y
E G E N S N G V L V L E R F L G C T T Q E A A D A V N D L L T S R V Q Q F E N T A L T E V P A L C L D K G L T P Q E C T A I A A Y
E G E N S N G V L V L E T F F G C G T Q Q A A E T V N D I L T S R L H Q F E D T A L T E V P A I A V E K G L T P G E V A A V A A Y
E G E L S N S V L V F E K F L G C S T Q E A A D A V N N L L T S R L H Q F E H T A V T E V P A L F D E H D V Q P A G R A E T L A Y
E G E N S N G V L V L E R F L G C S T Q E A A D A V N D L L T S R L Q Q F E N T A L T E L P P L C A E K G L S P D E T V A V M A Y
E G E L S N A V L V L E T F L G C T T Q Q A A E I S N D L I T S R L Q Q F E Q T A L T E L P R L F A E H A L A P A E I A A V L A Y
E G E L S N A V L V L E T F L G C T T Q E A A E A S N D L L T S R L Q Q F E Q T A L G E L P Q L F A D H A M D P A E I A A V L A Y
E G E L S N G V L V L E H F L G C T T P E A A E A V N D L L T S R L Q Q F E N T A L V E L P A L S A E Y G L D P A E N A A L A S Y
E G E L S N G V L V L E H F L G C T T P E A A E A V N D L L T S R L Q Q F E N T A L V E L P A L S A E Y G L D P A E N A A L A S Y
E G E L S N G V L V L E H F L G C T T P E A A E A V N D L L T S R L Q Q F E N T A L V E L P A L S A E Y G L D P A E N A A L A S Y
E G E L S N G V L V L E T F F G C T T Q E A A D T V N D V L T S R L H Q F E H T A L T E V P A V A L E N G L S P A E V T A V A R Y
E G E N S N G V L V L E K F L G C T T Q E A A N A V N D L L T S R L Q Q F E N T A L T E V P L L A A E K G L S A A E C A A V A A Y
E G E L S N G V L V L E T F L C C T T Q E A A D S V N E L L T S R L R Q F E D T T L T E L G P L F A E H G L D A A A C A G V L A Y
E G E N S N G V L V L E T F F G C T T Q E A A D T V N D V L T S R L H Q F E H T A F T E V P A V A L E K G L T P D E V V A V A A Y
E G E N S N A V L V F E R F F D C P T Q E A A E L V N D L L T S R L Q Q F E N T T L I E V P A L L A E N T V P V H E Q L G V A A Y
E G E L N N G V L V V E R F L G C S P Q R A A D L V N D I L T S R L H Q F E N T A L T E V P P L L A E H R I S P Q G R L D V L S Y
E G E N A N A V L V F E K F L G R S T Q E S A D L V N E L L T S R L H Q F E S T A L T E M P L L V A A K G V T P D Q Q A A V A A Y
E G E N S N C V L V L E R F L S C D T Q T A A D Y T N Q L L T S R L H Q F E Q T A L T E L A P L F A E H G V G P V E A V Q V G L Y

16
 Figure 1. Alignment of geosmin synthases.

401 411 421 431 441 451

YP_003098781 (DSM 43827)
AEK39836 (S699)
YP_003763541 (U32)
AEA03338 (CHAB 1432)
AEA03341 (CHAB 2155)
YP_716636 (ACN14a)
YP_483306 (Ccl3)
YP_001509819 (EAN1pec)
YP_004017381 (Eul1c)
YP_003265710 (DSM 14365)
BAJ30389 (KM-6054)
YP_634376 (DK 1622)
YP_001866236 (PCC 73102)
ZP_07114089 (PCC 6056)
ABU93239 (P2r)
ABU93238 (P2r)
YP_001105388 (NRRL 2338)
YP_001107098 (NRRL 2338)
YP_001106173 (NRRL 2338)
YP_001612078 (So ce 56)
ZP_01460669 (DW 4/3-1)
YP_003950745 (DW 4/3-1)
ZP_04705018 (J1074)
NP_823339 (MA-4680)
ADI05189 (BCW-1)
CCB75658 (NRRL 8057)
ZP_08240572 (XylebKG-1)
ZP_06769636 (ATCC 27064)
NP_630182 (A3(2))
ADW07414 (ATCC 33331)
ZP_04685148 (ATCC 14762)
ZP_08289513 (M045)
ZP_07309957 (Tü 4000)
YP_001828351 (NBRC 13350)
ZP_07296488 (ATCC 53653)
ZP_07293917 (ATCC 53653)
ZP_05522837 (TK 24)
ABY50951 (ATCC 27952)
ZP_06913794 (ATCC 25486)
YP_003487693 (87.22)
ZP_07276967 (AA4)
ZP_06273105 (ACTE)
ZP_07289987 (C)
ZP_05001875 (Mg1)
ZP_07976693 (SA3_actG)
ZP_07273435 (SPB78)
ZP_08453284 (Tü 6071)
ZP_06920565 (ATCC 29083)
CCA53556 (ATCC 10712)
ZP_07608000 (Tü 4113)
ZP_07307300 (DSM 40736)
YP_003335875 (DSM 43021)
YP_003382082 (DSM 17836)
YP_003509930 (DSM 44728)
YP_003116895 (DSM 44928)
Highly conserved residues are shown in red, deviations from otherwise highly conserved residues are shown in dark red. The strictly conserved motifs are shown in boxes. Added nucleotides of corrected sequences are shown in bold.
V K G L Q D W Q S G G H E W H M R S S R Y M N K G A A T S G H - P L L G G - - - - - P - - - - - - - - - - - - - - T - - - - - G L
V K G L Q D W Q S G G H E W H L R S S R Y M N E G A L T D R G - G P D L G G A - - - N - - - - - - - - - - - - - - - - - - - - G L
V K G L Q D W Q S G G H E W H L R S S R Y M N E G A L T D R G - G P D L G G A - - - N - - - - - - - - - - - - - - - - - - - - G L
I K G L Q D W Q S G G H E W H M R S S R Y M N K Q E P D N S G - T S V T L G G - - - P - - - - - - - - - - - - - - T - - - - - G L
I K G L Q D W Q S G G H E W H M R S S R Y M N K Q E P D N S G - T S V T L G G - - - P - - - - - - - - - - - - - - T - - - - - G L
I K G L Q D W Q S G G H E W H L R S S R Y M N R E A T P D A V - P P G L G P L A G L G G T G S L V P A A G L P G I P - - G I P S L
V K G L Q D W Q S G G H E W H L R S S R Y M N R A V A P E S G - E L S G L L G - - - L - - - - - - - - - - - - - - T - - - - - G L
V K G L Q D W Q A G G H E W H M R S S R Y M N A E S G A T G P - V P G S L P G D - - A - - - - - - - - - - - - - - T - - - - - G L
V K G L Q D W Q S G G H E W H L R S S R Y M N D G P R P D G I - A E T S Q R V A D R G I L P H - - - - - - - - - - - - - G P T G L
V K G L Q D W Q S G G H E W H M R S S R Y M N E G A L D D E G - G L P L G P - - - - T - - - - - - - - - - - - - - - - - - - - G L
V K G M Q D W Q S G G H E W H M R S S R Y M N G G G A A A P A - T G P T - - - - - - G - - - - - - - - - - - - - - - - - - - - - L
V R G L Q D W Q S G G H E W H M R S S R Y M N K G S G G A G G - F F L G P - - - - - N - - - - - - - - - - - - - - - - - - - - G L
I K G L Q D W Q S G G H E W H M R S S R Y M N K G G D N S P T - S T V L G G - - - - P - - - - - - - - - - - - - - T - - - - - G L
I K G L Q D W Q S G G H E W H M R S S R Y M N K E A D N S Q K - P P L F P G G - - - P - - - - - - - - - - - - - - T - - - - - G L
I K G L Q D W Q S G G H E W H M R S S R Y M N Q Q E D D N E V - K P T I L G W - - - P - - - - - - - - - - - - - - T - - - - - G L
I K G L Q D W Q S G G H E W H L R S S R Y M N E E A E T S P I - T G R L V I G - - - P - - - - - - - - - - - - - - T - - - - - G L
V K G L Q D W Q S G G H E W H I R S S R Y M N E G L V E Q S A L A G Q S A P G Q P A L P Q S A P D G T G P A T Q P V L G G P T G L
V K G L Q D W Q S G G H E W H L R S S R Y M N D G A L A G A R S P F G G P - - - - T - - - - - - - - - - - - - - - - - - - - - G L
V K G L Q D W Q S G A H E W H L R S G R Y A V P G G A E P R E P R R F L S G - - P - - - - - - - - - - - - - - H - - - - - - - G L
V K G L Q D W Q S G G H E W H M R S S R Y M N E N A D K S A D V P D F P G T - - - P - - - - - - - - - - - - - - T - - - - - - G L
V R G L Q D W Q S G G H E W H M R S S R Y M N K G A G G A G D T D G L P L G L - - S - - - - - - - - - - - - - - - - - - - - - G L
V R G L Q D W Q S G G H E W H M R S S R Y M N K G A G G A G D T D G L P L G L - - S - - - - - - - - - - - - - - - - - - - - - G L
V K G L Q D W Q S G G H E W H M R S S R Y M N E G A V D G E A A P P T P G G S P G A L A A A - - - - - - - - - - - - - L L G G T L
A R G L Q D W Q S G G H E W H L R S S R Y M N E G A L S Q K R P F G L S - - - - - A - - - - - - - - - - - - - - I - - - - - - - -
A K G L Q D W Q A G G H E W H M R S S R Y M N G G G A G D P G G P V P G G P L - - G - - - - - - - - - - - - - - L - - - - - - - -
V K G L Q D W Q S G G H E W H M R S S R Y M N G G A A P A A A T R W S P F A I - - G - - - - - - - - - - - - - - - - - - - - - G P
A K G L Q D W Q S G G H E W H M R S S R Y M N G G G A G E A V - - - - - - - - - - P - - - - - - - - - - - - - - - - - - - - - G F
T K G L Q D W Q S G G H E W H L R S S R Y M N E G L V D A G N P G G P - - - - - - I - - - - - - - - - - - - - - - - - - - - - - -
T K G L Q D W Q S G G H E W H M R S S R Y M N K G E R P L A G W Q A L T - - - - - G - - - - - - - - - - - - - - P - - - - - - - -
V K G L Q D W Q S G G H E W H M R S S R Y M N G G P S A T G T T S - - - - - - - - G - - - - - - - - - - - - - - F - - - - - - - -
A K G L Q D W Q S G G H E W H L R S S R Y M N Q G A R T T G P W A A P V G P G - - G - - - - - - - - - - - - - - P - - - - - - - -
A R G L Q D W Q S G G H E W H L R S S R Y M N E G A V A A R R P F G L P - - - - - A - - - - - - - - - - - - - - T - - - - - - - -
T K G L Q D W Q S G G H E W H M R S S R Y M N E G A R S G N P W G A P S - - - - - G - - - - - - - - - - - - - - P - - - - - - - -
A K G L Q D W Q S G G H E W H M R S S R Y M N G G G A G E A V P - - - - - - - - - G - - - - - - - - - - - - - - F - - - - - - - -
V K G L Q D W Q S G G H E W H M R S S R Y M N G G G V A A G E P A W S P F S V S G L G V G - - - - - - - - - - - G V - - - - S G L
V K G L Q D W Q S G G H E W H M R S S R Y M N G S G S E G S P A P S G L G G F S L L L S G - - - - - - - - - - - P T - - - - - G L
T K G L Q D W Q S G G H E W H M R S S R Y M N K G E R P L A G W Q A L T - - - - - G P - - - - - - - - - - - - - - - - - - - - - -
T K G L Q D W Q S G G H E W H M R S S R Y M N E G V E T E R S R F E G V - - - - - L - - - - - - - - - - - - - - A - - - - - - - -
T K G L Q D W Q S G G H E W H M R S S R Y M N D G V G E A V G P S S V A G - - - - V - - - - - - - - - - - - - - W - - - - - - - -
T K G L Q D W Q S G G H E W H M R S S R Y M N E G A T S A R G P L D L G G A V L S G P A L V - - - - - - - - - - T R - - - - A G H
V K G L Q D W Q S G G H E W H L R S S R Y M N K G A L D T L S G P E I L G G P - - T - - - - - - - - - - - - - - G - - - - - - - L
V K G L Q D W Q S G G H E W H M R S S R Y M N D G G A A Q A A P - - - - - - - - - G - - - - - - - - - - - - - - F - - - - - - - -
T K G L Q D W Q S G G H E W H M A S S R Y M N E G A R A T G R T T L P F L P T - - G - - - - - - - - - - - - - - L - - - - - - - -
A K G L Q D W Q S G G H E W H M V S S R Y M N K E A R P T A P L S L P F M P T - - G - - - - - - - - - - - - - - L - - - - - - - -
V K G L Q D W Q S G G H E W H L R S S R Y M N E G A V S G V A A G L T G V - - - - L - - - - - - - - - - - - - - - - - - - - - - -
V K G L Q D W Q S G G H E W H L R S S R Y M N E G A V S G V A A G L T G V - - - - L - - - - - - - - - - - - - - - - - - - - - - -
V K G L Q D W Q S G G H E W H L R S S R Y M N E G A V S G V A A G L T G V - - - - L - - - - - - - - - - - - - - - - - - - - - - -
T Q G L Q D W Q S G G H E W H M R S S R Y M N A R A D T T S P W Q A L T - - - - - G - - - - - - - - - - - - - - R - - - - - - - -
A K G L Q D W Q S G G H E W H M R S S R Y M N E G M V G G P S R L E G V - - - - - L - - - - - - - - - - - - - - - - - - - - - - -
V K G L Q D W Q S G G H E W H M C S S R Y M N G A N E A G G A H G A D E A G G T G Q A A E P G - A - - - - - - - W S P F A L S G L
T K G L Q D W Q S G G H E W H L R S S R Y M N E N A A R G S G P W P G F T - - G - - - - - - - - - - - - - - L - - - - - - - - - -
V K G L Q D W Q S G G H E W H A R S S R Y M N E G A A S G P A G V L R G P T - - - G - - - - - - - - - - - - - - L - - - - - - - -
V K G L Q D W Q S G G H E W H L R S S R Y M N D G G Q A A T L - G P S A L L D - - - - - - G - - - - - - - - - - - - - - - P K G L
V K G L Q D W Q S G G H E W H M R S S R Y M N E S A R A V P L G P - - - - - - - - M - - - - - - - - - - - - - - - - - - - - - G L
I K G L Q D W Q A G G H E W H M R S S R Y M N D A A D G T G A - G T S A G G A S A A L A G - - - - - - - - - - - - P - - - - L G L

17
 Figure 1. Alignment of geosmin synthases.

461 471 481 491 501 511 521

YP_003098781 (DSM 43827)
AEK39836 (S699)
YP_003763541 (U32)
AEA03338 (CHAB 1432)
AEA03341 (CHAB 2155)
YP_716636 (ACN14a)
YP_483306 (Ccl3)
YP_001509819 (EAN1pec)
YP_004017381 (Eul1c)
YP_003265710 (DSM 14365)
BAJ30389 (KM-6054)
YP_634376 (DK 1622)
YP_001866236 (PCC 73102)
ZP_07114089 (PCC 6056)
ABU93239 (P2r)
ABU93238 (P2r)
YP_001105388 (NRRL 2338)
YP_001107098 (NRRL 2338)
YP_001106173 (NRRL 2338)
YP_001612078 (So ce 56)
ZP_01460669 (DW 4/3-1)
YP_003950745 (DW 4/3-1)
ZP_04705018 (J1074)
NP_823339 (MA-4680)
ADI05189 (BCW-1)
CCB75658 (NRRL 8057)
ZP_08240572 (XylebKG-1)
ZP_06769636 (ATCC 27064)
NP_630182 (A3(2))
ADW07414 (ATCC 33331)
ZP_04685148 (ATCC 14762)
ZP_08289513 (M045)
ZP_07309957 (Tü 4000)
YP_001828351 (NBRC 13350)
ZP_07296488 (ATCC 53653)
ZP_07293917 (ATCC 53653)
ZP_05522837 (TK 24)
ABY50951 (ATCC 27952)
ZP_06913794 (ATCC 25486)
YP_003487693 (87.22)
ZP_07276967 (AA4)
ZP_06273105 (ACTE)
ZP_07289987 (C)
ZP_05001875 (Mg1)
ZP_07976693 (SA3_actG)
ZP_07273435 (SPB78)
ZP_08453284 (Tü 6071)
ZP_06920565 (ATCC 29083)
CCA53556 (ATCC 10712)
ZP_07608000 (Tü 4113)
ZP_07307300 (DSM 40736)
YP_003335875 (DSM 43021)
YP_003382082 (DSM 17836)
YP_003509930 (DSM 44728)
YP_003116895 (DSM 44928)
Highly conserved residues are shown in red, deviations from otherwise highly conserved residues are shown in dark red. The strictly conserved motifs are shown in boxes. Added nucleotides of corrected sequences are shown in bold.
G T S - - E L R S I L R T L P Q - - - - - - - - - - - - R A R S F T H R P F A E V P P R G R - - P N T R L P Y R V E L S P H W R S
G T S A A R I F A S V L A T A P Q - - - - - - - - - - - R L R A Y G S T P F R A V D V P R - - - P S V D V P F P L R L S P H L A A
G T S A A R I F A S V L A T A P Q - - - - - - - - - - - R L R A Y G S T P F R A V D V P R - - - P S V D V P F P L R L S P H L A A
G T S A A R L E S L S T T L G L R - - - - - - - - - - - R F K S F T H V P Y Q T V G P V K L - - P K F Y M P F S T T L N P N L D A
G T S A A R L E S L S T T L G L R - - - - - - - - - - - R F K S F T H V P Y Q T V G P V K L - - P K F Y M P F S T T L N P N L D A
G T S A I Q V L P S L L A T A P R - - - - - - - - - - - R I R S F A N V P F R L V G P T P L - - P E F Y L P Y T T G L S P H L D S
G T S A A R I V P S L V T T T P R - - - - - - - - - - - R I R S F T H I P H Q I V G P L R H - - P D F C M P F S T G Q S P H L D A
G T S A V R I A A S L L A T A P A - - - - - - - - - - - R M R A F T H V P H Q V V G P V K L - - P A F Y M P F T T G E S R H L A A
G S S I A R P L E S V L A T A P Q - - - - - - - - - - - R G R A F S H V P F Q V V G P S R A - - P A P Y M P F P V G D N P H L P G
G T G S L R Q I A A T F K Q G L R - - - - - - - - - - - R G K R Y S H P P F R R V G P V P L - - P D F Y M P Y S T T L N P H L D G
G T S A A N I R L T A G L R G - - - - - - - L - - - - - - - R G L A H T P Y R K T G P A L L - - P D F P M P F P L S I N P H L D A
G T S A A R L P Q S P T A L G L T - - - - - - - - - - - R L K N F S H V P Y Q P V G P V K L - - P K F Y M P Y S T K P S P H L D A
G T S A A R I E S L Y A A L G L G - - - - - - - - - - - R I K S F T H V P Y Q P V G P V T L - - P K F Y M P F T T S L N P H L N A
G T S A A R I E S L Y T T L G L G - - - - - - - - - - - R F K S F T H V P Y Q A V G P V K L - - P K F Y M P F S T S L N P N L D A
G T S G I R F E S L Y A T L G L G - - - - - - - - - - - R F N S F T H V P Y Q A V G P V K L - - P K F Y M P F S T T L N P N L D A
G T S L A R I F D L P R D T K R P G - - - - G - - - - - - L K T E K Q V K D D F N Y E F E F - - P N F Y M P F S A Q V N P H L E A
G T S A A R I V Q S L L S T A P Q - - - - - - - - - - - R I R S F T H T P Y E P A G P I R M - - P E I Y M P F D L S L S P H L D V
G T S A A H N A L A R V R P G I R - - - - - R - - - - - H R E Q H S H A P F A P V G H L P L - - P E I Y M P F P V R M S P H L D A
G T S S S H L G S L L R T - - - - - - - - V - - - - - - - - R P G L P I P H G Q L R Y A R I A V P A M S S P H P V R T N P Q V G T
G T S A F R I R L T P A A L G L Q - - - - - - - - - - - R I K T L T H P P F Q P V G R M P L - - P K I S M P F K L R L S P H L S A
G L S A V R F P F S A S A L G L N - - - - - - - - - - - R F K S F T H T P Y M P V G P V K L - - P K F Y M P Y S T S V S P H L D A
G L S A V R F P F S A S A L G L N - - - - - - - - - - - R F K S F T H T P Y M P V G P V K L - - P K F Y M P Y S T S V S P H L D A
G T S A L D V R A L F G R P A R Q - - - - - - - - - - - R A R S F S H V P Y Q R T G A S L L - - P D F K L P F P T R L S P A L E Q
G T S A A D L R G L L A D A G A E - - - - - - - - - - - R L R R Y T H V P F Q K V G P S R I - - P D F H M P F Q V E L S P H L E G
G T S A A R I A T S L A A T L P F - - - - - - - - - - - R L R S H A Q P P R R V V G P V R I - - P D L H M P F P A R L S P H L D R
G T S A A D L R L T T G R V G A G - - - - - - - - - - - R V R A F T H V P H Q R V G P S R L - - P A F H M P F P T T L N P H L P T
G M S A A S I R F T L R S E T A - - - - - - - - - - - - R A R S L S H V P F Q Y V G P S L L - - P D F D L P F G T T L S P H L A G
G I S A L D I R T L F G R P A A Q - - - - - - - - - - - R L R S L T H L P H Q R V D S R V - - - P D F E L P F P L S L S P Y R H G
G T S A A D V G A L L A D A V A Q - - - - - - - - - - - R A R S Y T Y V P F Q K V G P S V I - - P D I R M P Y P L E L S P A L D G
G M A A A S V R L T P R S E S A - - - - - - - - - - - - R L R S L T H V P Y R H V G P S L L - - P D F E M P F T T T L S P H L G G
G T S A A D V R A L L A A P G A P G A P S A P - - - W R R T R A H T H V P Y Q K V G P S L I - - P D I R M P F P L A L S P H L D A
G T S A A D V R G L L A D A G A E - - - - - - - - - - - R L R A Y T H V P F Q K V G P S R I - - P E M R M P F T L R L S E H L D G
G T T A A D V G A L L A G A A T E - - - - - - - - - - - R L R A H A H V P Y R K V G P S V I - - P E I R M P F P L R L S P A L D G
G M S A A S I R F T L R S E T A - - - - - - - - - - - - R A R S L S H V P F Q Y V G P S L L - - P D F D L P F G T T L S P H L A G
G V S A A D I R L T T G R A E R A - - - - - - - - - - - R A R G F S H V P H Q R V G P S R L - - P D F F M P F T A R L S P H L D R
G T S A A R I G L T T T R R A E S G - - - - - - - - - - R L R S F T H V P H Q R V G P S Q L - - P D F F M P F T T R L S P H L D G
G T S A A D V G A L L A D A V A Q - - - - - - - - - - - R A R S Y T Y V P F Q K V G P S V I - - P D I R M P F P L E L S P A L D G
- T S A L D I R T L F G R P A A A - - - - - - - - - - - R M R T L T H R P - Q Q V G P S W L - - P D F D L P F P L S L S P H L E Q
G T S A L D I R T L F G R P A A A - - - - - - - - - - - R L R T L T H R P R Q - V G P S L L - - P D F A L P F P L T L S P H L D D
G T S A A D V G A L L A T A A A Q - - - - - - - - - - - R L R A H T H Q P Y Q K V G P S L L - - P D F H M P F R V A L C P H L D G
G T S A A R I F R S V T A T A P Q - - - - - - - - - - - R T R A F T H Q P H E Q L G P V A L - - P S L P L P F R L Q L S P H L A A
A M A A A S I R F T P R S E S A - - - - - - - - - - - - R L R S H T H V P Y Q H V G P S L L - - P D F E M P F S T T L S P Y L D G
G T S A F D L R S A F T R R S M E L - - - - - - - - - - R R R S F T H V P H Q K T G P L L L - - P D I R M P Y E L R L S P H L D H
G G A A L D L R S V L E P R A L E R - - - - - - - - - - R R R S F S H V P F E R T G P S V I - - P D I H M P F P L S L S P H H A H
G T S A V H L A A A L G T P A R A - - - - - - - - - - - R L R S H T R P P H T H E G P S L L - - P D F T L S Y P L G L S P Y H E R
G T S A V H L A A A L G T P A R A - - - - - - - - - - - R L R S H T R P P H T H E G P S L L - - P D F T L S Y P L G L S P Y H E R
G T S A V H L A A A L G T P A R A - - - - - - - - - - - R L R S H T R P P H T H E G P S L L - - P D L T L S Y P L G L S P Y H E R
G T S A A D V G A L L A A A G A E - - - - - - - - - - - R L R A H T H V P F Q K V G P S L L - - P D F Y M P F Q V E H S P H L P G
G T S A F D I R T L F G R P A A S - - - - - - - - - - - R L R S L T H V P H Q R V G P S L L - - P E F D L P Y P L A L S P H H A E
G V S A A S L P L T T G R A E A A - - - - - - - - - - - R A R T F S H V P F Q K T G P S L L - - P D F F M P F P L R L N A H L P T
G T S A A D V R A L L A T A G A M G A P P T P F G Q W G R L R S Y T H V P Y Q K V G P A K I - - P D I R M P F P L E L S P H L D D
G T S A A V P T L S P A R L G L R R - - - - - - - - - - R S Q Q Q S H R P F Q P V G H L P L - - P D L Y M P Y P V R T S P H L D A
G T A A S R I L Q S M V T S A P Y - - - - - - - - - - - R I R S Y T H R L Y D V V E P F E R - - P E L Y M P Y R A K L S P H L D R
G A D S L R A G V S - - - - - - - - - - - - G - - - - - - L G N H S R V P F Q K V G K Q R L - - P D M Y M P F Q V Q L N P A L D A
G T S A A R I L A S I A K T M P A - - - - - - - - - - - R L K R V S H P P H E Q V G P T P I - - P A I A A P S E L R L S P H L G L

18
 Figure 1. Alignment of geosmin synthases.

531 541 551 561 571 581

YP_003098781 (DSM 43827)
AEK39836 (S699)
YP_003763541 (U32)
AEA03338 (CHAB 1432)
AEA03341 (CHAB 2155)
YP_716636 (ACN14a)
YP_483306 (Ccl3)
YP_001509819 (EAN1pec)
YP_004017381 (Eul1c)
YP_003265710 (DSM 14365)
BAJ30389 (KM-6054)
YP_634376 (DK 1622)
YP_001866236 (PCC 73102)
ZP_07114089 (PCC 6056)
ABU93239 (P2r)
ABU93238 (P2r)
YP_001105388 (NRRL 2338)
YP_001107098 (NRRL 2338)
YP_001106173 (NRRL 2338)
YP_001612078 (So ce 56)
ZP_01460669 (DW 4/3-1)
YP_003950745 (DW 4/3-1)
ZP_04705018 (J1074)
NP_823339 (MA-4680)
ADI05189 (BCW-1)
CCB75658 (NRRL 8057)
ZP_08240572 (XylebKG-1)
ZP_06769636 (ATCC 27064)
NP_630182 (A3(2))
ADW07414 (ATCC 33331)
ZP_04685148 (ATCC 14762)
ZP_08289513 (M045)
ZP_07309957 (Tü 4000)
YP_001828351 (NBRC 13350)
ZP_07296488 (ATCC 53653)
ZP_07293917 (ATCC 53653)
ZP_05522837 (TK 24)
ABY50951 (ATCC 27952)
ZP_06913794 (ATCC 25486)
YP_003487693 (87.22)
ZP_07276967 (AA4)
ZP_06273105 (ACTE)
ZP_07289987 (C)
ZP_05001875 (Mg1)
ZP_07976693 (SA3_actG)
ZP_07273435 (SPB78)
ZP_08453284 (Tü 6071)
ZP_06920565 (ATCC 29083)
CCA53556 (ATCC 10712)
ZP_07608000 (Tü 4113)
ZP_07307300 (DSM 40736)
YP_003335875 (DSM 43021)
YP_003382082 (DSM 17836)
YP_003509930 (DSM 44728)
YP_003116895 (DSM 44928)
Highly conserved residues are shown in red, deviations from otherwise highly conserved residues are shown in dark red. The strictly conserved motifs are shown in boxes. Added nucleotides of corrected sequences are shown in bold.
A L E H N V E W V G R M G V Y D D S P - - - - - A V W T A R K V R D Y D F A L C S A G L D P D A T A E A L N L S A D W L A F G T Y
C R I R N V D W A R R T G F L D - - - - - - - G V V W D E R K L R A A D L P L C A A G I H P D A T A E G L D V T S D W L T W G T Y
C R I R N V D W A R R T G F L D - - - - - - - G V V W D E R K L R A A D L P L C A A G I H P D A T A E G L D V T S D W L T W G T Y
A R N H S K E W A R R M G M L E S L P E I P D A F I W N D H K F D V A D V A L C G A W I H P N G S E H E L N L T A C W L V W G T Y
A R N H S K E W A R R M G M L E S L P E I P D A F I W N D H K F D V A D V A L C G A W I H P N G S E H E L N L T A C W L V W G T Y
S R R A I I P W A R S M G M L D R V P - - - - - G I W D E H K L W S Y D F A L C S A G I H P D A T A D E L D L T T A W L T W G T Y
S R R E N I I W A R A V G M L D P I P - - - - - G I W D E H K L R A F D F A L C S A G I H P D A T L P E L N L T T D W L T W G T Y
A R H N I V E W S A A V G F L D P V P - - - - - G I W D E H K L R A A D F A L C S A A I H P N A T A A E L D L T T G W L T W G T Y
C R R D S V R W A R D M G L L A Q V P G - - - - - I W D E H K L V S Y D F P L C S A G L D P D A S Q A D L L L S A C W L T W G T Y
S R R F S K E W A R S M G L L D V V P E V P G G Y I W D E H K F D V A D V A L C G A L I H P E A S A A E L N V T A C W L V W G T Y
A R A Y V V G W A R S T G L L D P E P G V P A S D I W T E A K L R D Y D F P L C A A G I D P D G T P A E L D L S S A W L T W G T Y
A R R D S K A W A R R M G M L D V L P G V P G G Y I W D D H K F D V A D V A L C G A L I H P H A T A A Q L N L S S C W L V W G T Y
A R K H S K E W A R Q M G M L E S L P G I P D A V I W D D H K F D V A D V A L C G A L I H P N G S G L E L N L T A C W L V W G T Y
A R K H S K E W A R Q M G M L E S L P G I P D A F I W N D H K F D V A D V A L C G A L I H P H G S E H E L N L S A C W L V W G T Y
A R K H S K E W A R Q M G M L A T V P G I P D A F I W N D H K F D V A D V A L C G A W I H P N G S E H E L N L T A C W L V W G T Y
V R L H I K A W A I A M G M L S P G E D S L N L G I W D E R K F D L M N L A F F A S V T N P D L T I I Q L E I V A D W C V W M F F
C R E N T A A W A R A M G I F D D V P - - - - - R V W D E N Q M R G Y D L P L C S A G L D P D A T P E E L D L S A A W L T W G T Y
A R Q H A V D W A R E M G M F D S V P G S E V G G V W N E R R F V G F D F P H C A A M I H A D A G P E Q L D L S S D W L A W G T Y
V R A H A K E W A R R M G M L D - - - - - - G S G V W T A N V F D A A D F G Q F S A M A H P D S P G P E L E L V N D W H V W G W F
A R R S T L E W A R R F A M L D V V P G V F G S G I W D E R R F I G F D F P I C A A G I H P D A S P H E L D L S S A W L T W G T Y
A R R H S K E W A R Q M G M L D S L P G L P G V Y I W D D H K F D V A D V A L C G A L I H P E A S A E Q L N L T A C W L V W G T Y
A R R H S K E W A R Q M G M L D S L P G L P G V Y I W D D H K F D V A D V A L C G A L I H P E A S A E Q L N L T A C W L V W G T Y
S R V D T V E W A H R M G L L V P Q P G V P G S D V W D E E A L A G Y D F P L C A A G L H P E A D L E E L N L A S Q W L T W G T Y
A R A R L T P W M H S T G M L Q - - - - - - - E G V W D E D K L T A Y D L P L C S A G L D P D A T P D E L D L S S R W L A W G T Y
S R E H I M E W G W R M G M L D V P P G L P G F G V W D E H K L R A F D F A L A A A G I H P D A T G E E L D L T T G W L T W G T Y
A R R A T V A W G H R M G M L E P L P E L L G G H L W D E H K L H A F D F P L C A A G I N P D A P P E A L D L A S Q W L T W G T Y
A R G R L V D W A R R M G I L E A Q P G V P G S H I W D E R R I A A I D L P L C A A G I H P D A S P D E L D L S S G W L A W G T Y
A L E Q S I A W A R Q M G L L D - - - - - - - - G L W D E T M L R G F D F A L C A A G I D P D A T P E E L E L S T Q W M T W G T Y
A R R H L S E W C R E M G I L S - - - - - - - E G V W D E D K L E S C D L P L C A A G L D P D A T Q D Q L D L A S G W L A F G T Y
A R V R I V E W S R R M G L L E A Q P G V P G S Y I W D E A R L I A T D L P L C A A G L H P D A T P E E L D L S S G W L T W G T Y
A R A H L V D W C H G T G I L H - - - - - - - E G V W D E D K L T A Y D L A L C S A G L D P D A T P E A L D L S A Q W L A F G T Y
A R A R L L P W T R E A G M L S - - - - - - - E G V W D E D K L A S C D L P L C A A G L H P D A T P E A L D L A S Q W L A W G T Y
A R S H L L E W S R R M G I L - - - - - - - G E G V W D E D K L E S C D L A L C A A G L D P D A T Q D E L D L A S G W L A F G T Y
A R V R L V D W A R R M G I L E A Q P G V P G S H I W D E R R I A A I D L P L C A A G I H P D A T P D E L D L S S G W L A W G T Y
A R R N L I E W S H R M G L F G P Q P G A L D S C V W D E H R L L A A D L P L C A A G I H P G A S P D E L D L T S G W L T W G T Y
A R R N A V E W G R R M G M L E A Q P G I P G S H I W T A R K L A G F D F P L C A A G I H P D A T P D Q L D I T S G W L T W G T Y
A R R H L S E W C R E M G I L S - - - - - - - E G V W D E D K L E S C D L P L C A A G L D P D A T Q D Q L D L A S G W L A F G T Y
A R A A S V A W A G R M G L L G - - - - - - - - D I W D E A K L T G F D F A L C S A G L D P D A T P E E L E L S A E W L T W G T Y
A R R K S V D W A G R M G L L N - - - - - - - - D I W D E A K I K G F D L A L C A A G L D P D A T P E E L E L S A E W L T W G T Y
A R P R L T A W A H A M G I L S - - - - - - - E G V W D E E R L A A A D L P L C S A G L D P D A T P E Q L D L S S A W L A W G T Y
E R K K I T D W P R R M G F L S - - - - - - - E G V W D A T R L H K I D I P V A S A G L C P A A P P E Q L S L T T D W L T W G T Y
A R V R I V D W S R R M G L L E A Q P G V P G S H I W D E E R L I A T D L P L C A A G L H P D A T P E E L D L S S G W L T W G T Y
A R E G S L S W A R R T G L L D A R P G D P G S A I W D E Q K V R G Y D F A L C S A A I A P D A T A G A L L L N A C W L T W G T Y
A R E G S V A W A R E M G L L D P Q P G D P G S A I W N E P K L R G Y D F A L C S A G I D P D A T R E A L L L N A C W L T W G T Y
A R T A S V D W A E R M G L L N - - - - - - - - D I W D R E K L E G Y D F A L C S A G L D P D A S P E E L E L S A E W L T W G T Y
A R T A S V D W A E R M G L L N - - - - - - - - D I W D R E K L E G Y D F A L C S A G L D P D A S P E E L E L S A E W L T W G T Y
A R T A S V D W A E R M G L L N - - - - - - - - D I W D R E K L E G Y D F A L C S A G L D P D A S P E E L E L S A E W L T W G T Y
A R P R L V E W A H R M G M L Q - - - - - - - E G V W D E D K L A A A D L P L C S A G I D P D A S P E A L D L S S H W L A W G T Y
A G R L S L D W A E R F G L L D - - - - - - - - D L W D R P M A E G F D L A L C S A G L D P D A T L E E L E L S T E W L T W G T Y
A R R H L T D W A H R M G I L E P Q P G L P G S Q V W D E E R L L A A D L S L C A A G I H P D G S L D E L D L V S G W L A W G T Y
A R R R L R D W V G R M G I L A - - - - - - - E G V W D E D K L R A Y D L A L C S A G L D P D A T P E A L G L S A Q W L A W G T Y
A R R Y A V G W A R R M G M F D A I P G V E A G G L W D E R R F I G F D F A H C A A M I H A D A S P E Q L N L S S D W L A W G T Y
A R A N V L E W G Q R M G L T D - - - - - - - G V I W D A R M L L A N D L P L C A A G I H P D A D P D A L D L S S Q W L A W G T Y
A R E H N V E W S R E M G F L S P I P G V P G P G V W N E R Q I R V F D L A L C A A G I D P D G T P E E L E L S A D W L A W G T Y
A A R H T V D W S R R I G F F D - - - - - - - - G I W D E Q H L L S C D L A L C A A G I S P D A T P D E L N L A T D W L S W G T Y

19
 Figure 1. Alignment of geosmin synthases.

591 601 611 621 631 641 651

YP_003098781 (DSM 43827)
AEK39836 (S699)
YP_003763541 (U32)
AEA03338 (CHAB 1432)
AEA03341 (CHAB 2155)
YP_716636 (ACN14a)
YP_483306 (Ccl3)
YP_001509819 (EAN1pec)
YP_004017381 (Eul1c)
YP_003265710 (DSM 14365)
BAJ30389 (KM-6054)
YP_634376 (DK 1622)
YP_001866236 (PCC 73102)
ZP_07114089 (PCC 6056)
ABU93239 (P2r)
ABU93238 (P2r)
YP_001105388 (NRRL 2338)
YP_001107098 (NRRL 2338)
YP_001106173 (NRRL 2338)
YP_001612078 (So ce 56)
ZP_01460669 (DW 4/3-1)
YP_003950745 (DW 4/3-1)
ZP_04705018 (J1074)
NP_823339 (MA-4680)
ADI05189 (BCW-1)
CCB75658 (NRRL 8057)
ZP_08240572 (XylebKG-1)
ZP_06769636 (ATCC 27064)
NP_630182 (A3(2))
ADW07414 (ATCC 33331)
ZP_04685148 (ATCC 14762)
ZP_08289513 (M045)
ZP_07309957 (Tü 4000)
YP_001828351 (NBRC 13350)
ZP_07296488 (ATCC 53653)
ZP_07293917 (ATCC 53653)
ZP_05522837 (TK 24)
ABY50951 (ATCC 27952)
ZP_06913794 (ATCC 25486)
YP_003487693 (87.22)
ZP_07276967 (AA4)
ZP_06273105 (ACTE)
ZP_07289987 (C)
ZP_05001875 (Mg1)
ZP_07976693 (SA3_actG)
ZP_07273435 (SPB78)
ZP_08453284 (Tü 6071)
ZP_06920565 (ATCC 29083)
CCA53556 (ATCC 10712)
ZP_07608000 (Tü 4113)
ZP_07307300 (DSM 40736)
YP_003335875 (DSM 43021)
YP_003382082 (DSM 17836)
YP_003509930 (DSM 44728)
YP_003116895 (DSM 44928)
Highly conserved residues are shown in red, deviations from otherwise highly conserved residues are shown in dark red. The strictly conserved motifs are shown in boxes. Added nucleotides of corrected sequences are shown in bold.
G D D Y Y P Y V F G R T P G M A A P K A L T E R L K Q L M P V E - - - - G P A P I S - P A S P L E L M L A D L W A R T T E P M D V
A D D Y Y P V E F G A T R D L A G A K A A N L R L S A F M P V H - - - D S P M P A - - P A T A L E A G L A D L W P R T T A G M P T
A D D Y Y P V E F G A T R D L A G A K A A N L R L S A F M P V H - - - D S P M P A - - P A T A L E A G L A D L W P R T T A G M P T
A D D Y F P A I Y G N N R D L A G A K V F N A R L S A F M P L D - - - - N S T P P V - A T N P V E K G L A D I W S R T A G P M S S
A D D Y F P A I Y G N N R D L A G A K V F N A R L S A F M P L D - - - - N S T P P V - A T N P V E K G L A D I W S R T A G P M S S
G D D Y Y P V I F G A S R N L A A A K L C N E R L R L F M P V D - - - G P L T E P - - P V N A L E R G L A D L W E R T G A G M E P
A D D Y Y P V I F G R T R D I L G A K V C N A R L S E F M P L D - - - - - S P V T A V P A N A L E R G L A D L W T R T T E T M A P
A D D L Y P V L Y G R T R D L A G A R A C T E R L K E L M P V E - - - - - P G P L P V P V G G L E R G L A D L W P R T T R D M T P
A D D Y Y P V I F G R A R D L A G A R A C N E R L R Q F L P V E F A T L A G A T P P - P L L A L E R G L A D L W A R T A P P L S L
A D D Y F P A L F G D R R D W L G A K L S V S R L S A F M P L E A A - E L A N P S A V P V G P L E H S L A D L W A R S A P G L S S
A D D Y Y P A V Y G R P R D L L G A K A Q G E R L H A L M T L D - - - - - G T L P F A P R N A L E T G L A D L W R R T V A P F D Q
A D D Y F P A F Y G H T K D M A G A K V F N A R L A L F V P E D A - - G A V V P P - - P T N P V E R G L A D L W A R T T E G V T P
A D D Y F P A L Y G N N R N M A G A K V F N A R L S A F M P L D - - - - D S T P S E V P T N P V E A G L A D I W S R T A G P M S A
A D D Y F P A L Y G N N R D M V G A K V F N A R L S A F M P L D - - - - D S T P P A V P T N P V E R G L A D I W S R T A G P M S S
A D D Y F P A I Y G N I R D L A G A K V F N A R L G A F M P L D - - - - D S T P P A V P T N P V E K G L A D I W S R T A G P M S S
E D D Y F H E R Y K R T R D L V G A K E F I K R I P A F M P V D - - - - - L T P P P V P T N P L E R A L I D L W P R T A S H L P L
G D D Y Y P R V F G R T L D M A G A R A C N A R L K E L M P V E - - - - - S A P A T A P V T P L E R G L A D L W A R T A G P M P V
G D D F F P V V F G A T R N L A A A K V C N D R L S A F M P I D - - - G G G V P E - - P A N V L E R G L A D L W R R T A G P M P A
F D D F F T E V F K R S R N R A G A E A F L A R L P G F M P A D T - - - R R T P A - - P A N P V E R G L A D L W A R S T P V L A P
G D D Y F P R V F G A T R D M A G A K R F N A R L S M F M P L D - - - - - C A G A P V P E N P V E S G L L D L W I R T A S P M S M
A D D Y F P A F Y G Y T R D M A G A K L F N A R L S A F M P - D - - - - - G P C T A V P T N P V E H G L A D L W A R T A G P M T D
A D D Y F P A F Y G Y T R D M A G A K L F N A R L S A F M P - D - - - - - G P C T A V P T N P V E H G L A D L W A R T A G P M T D
G D D Y Y P A V F G R A R N F A G A K L C T D R L R D F M P V E - - - - Q P S A G P V P T S A L E R S L A D L W V R T A S A M P E
G D D Y Y P M V F G P R R D L A A A K L C T R R L S A C M P V D - - - G E E V P A - - P V N G M E R G L I D L W A I T T A E M T P
A D D Y Y P M V F G R A F D L V G A R V C N E R L S A F M P V D - - - - - S T A V P T P V T A L E R G L A D L W S R T A G P M T V
G D D Y F P V V F G R T R D L A G A K A C N D R L S A F M P I G - - - A S A T P P - - P A T P L E R G L A D L W A R T A G P M T T
G D D W F P V V H G R T R D L A G A R L A N E R L S L F M P L D - - - G G G T P E - - P V N A L E R S L G D L W R R T A A P M D L
T D D W Y P A V F G R S R D L A G A K L C H E R L L A C V P L H - - - D P A A G A H A A G T P M E R A L A D L W A R T A G P M G M
G D D Y Y P L V Y G H R R D L A A A R L T T T R L S D C M P L D - - - G E P V P P - - P G N A M E R S L I D L W V R T T A G M T P
G D D W F P V V H G R S R D L A G A R V A N E R L S L F M P L D - - - G A S A P E - - P A N A L E R G L A D L W R R T A A P M D G
G D D Y Y P L V Y G H R R D L A A A R L T T A R L A D C M P V D - - - G E P V P P - - P A N A M E R A L T D L W A R T T A A M T P
G D D Y Y P L M Y G H R R D L A G A R L C T E R L S A C M P V D - - - - - G E P T L L P V N A L E R S L I D L W Q R T T A G M D R
G D D Y Y P L V Y G H R R D L A A A R L T T A R L S A C M P L D - - - G E E V P P - - P V N A M E R S L T D L W T R T T A G M T P
G D D W F P V V H G R T R D L A G A R L A N E R L S L F M P L D - - - G G G T P E - - P V N A L E R S L G D L W R R T A A P M D L
A D D Y Y P Q V F G R R R D L T G A K A C N E R L K T F M P V D - - - - - A G P A P V P A G G L E R S L A D L W A R T A G P M E E
G D D Y F P V V F G R T R D L A G A K A A N Q R L S S F M P V D - - - - - G S A P P A P A N A L E R G L A D L W T R T A G P M T P
G D D Y Y P L V Y G H R R D L A A A R L T T T R L S D C M P L D - - - G E P V P P - - P G N A M E R S L I D L W V R T T A G M T P
G D D Y Y P L V F G R A R A L E G A R L C N E R L K A C M P V D - - - E P A A G A A V A V A P M E R S L A D L W A R T A G P M S P
G D D Y Y P V V F G A P R D L A G A R L C N D R L K A C M P V D - - - A P E A G A A V A V A P M E R S L A D L W A R T A G P M S A
G D D Y Y P L V F G H R R D L A A A R L T T A R L S D C M P L D - - - - - G E R A P L P S N A M E R A L V D L W T R T T A A M T P
A D D Y Y T E A L S H - - D L S A A R L T T E R L K L F M P T D - - - - - G K P P P T P Q T A L E T G L A N L W Q R T T G P L S P
G D D W F P V V H G R S R D L A G A R L A N E R L S L F M P L D - - - - - G T S V P E P V N A L E R G L A D L W R R T A G P M D E
A D D Y Y P V V F G R T G D I A A A K A A T A R L V A M M P S Q - - - - A G E A G P A P V T V L E R G L A D L W A R T V R G M G A
G D D Y Y P V V F A Q T K N L P A A K A T T A R L I A M I P L D - - - - - H T E R P E P A T A M E R A L G D L W V R T S A Q M G P
G D D Y Y P L V F G R P R D L V A A R V L H E R L L A C M P L E - - - D P A L A A A F A A A P I E R S L A D L W T R T A G P M D P
G D D Y Y P L V F G R P R D L V A A R V L H E R L L A C M P L E - - - D P A L A A A F A A A P I E R S L A D L W T R T A G P M D P
G D D Y Y P L V F G R P R D L V A A R V L H E R L L A C M P L E - - - D P A L A A S F A A A P I E R S L A D L W T R T A G P M D P
G D D Y Y P L V Y G G R R D L A A A R L T T Q R L S D C M P L D - - - - - G E Q T L V P V N A M E R G L I D L W A R T T A E M T P
G D D Y Y P A V F G R T R N L L G A K E Q T E R F K A C M P L D - - - D P A A G A A L A V N P M E R S L A D L W A R T A G P Q S P
A D D Y Y P A V F G R T H D L A G A R A C N A R L G A F M P L D - - - - - A G P T P A P V T A L E G S L A D L W A R T A G P M E D
G D D Y Y P L V H G H R R D L A A A R L T T A R L S A C M P L A G - - - - - E E P P V P A N A M E R G L V D L W L R T T A G M T P
G D D Y F P A V F G A P R D L V A A K L C N E R L S A F M P L D - - - - - A G A T P E P T N P I E R G L E D L W R R T A E P M S V
A D D Y Y P V A F G R S R D L V G A K A T N K R L S A L M P L E - - - L D A A G P P V P G N A L E R S L A D L W S R T A G P M A P
G D D L Y P T V Y G R A K D L A G A Y A Q N Q R L V R F M P V E - - - G E E A P Q - - A V N A L E R G L A D L W R R T V Q P L D D
A D D Y Y P A V F G P T A D R V G A K L S N E R L S A L M A - D - - - - - N P P A - - P V S A L E R G L A D L W R R S T A D A S P

20
 Figure 1. Alignment of geosmin synthases.

661 671 681 691 701 711

YP_003098781 (DSM 43827)
AEK39836 (S699)
YP_003763541 (U32)
AEA03338 (CHAB 1432)
AEA03341 (CHAB 2155)
YP_716636 (ACN14a)
YP_483306 (Ccl3)
YP_001509819 (EAN1pec)
YP_004017381 (Eul1c)
YP_003265710 (DSM 14365)
BAJ30389 (KM-6054)
YP_634376 (DK 1622)
YP_001866236 (PCC 73102)
ZP_07114089 (PCC 6056)
ABU93239 (P2r)
ABU93238 (P2r)
YP_001105388 (NRRL 2338)
YP_001107098 (NRRL 2338)
YP_001106173 (NRRL 2338)
YP_001612078 (So ce 56)
ZP_01460669 (DW 4/3-1)
YP_003950745 (DW 4/3-1)
ZP_04705018 (J1074)
NP_823339 (MA-4680)
ADI05189 (BCW-1)
CCB75658 (NRRL 8057)
ZP_08240572 (XylebKG-1)
ZP_06769636 (ATCC 27064)
NP_630182 (A3(2))
ADW07414 (ATCC 33331)
ZP_04685148 (ATCC 14762)
ZP_08289513 (M045)
ZP_07309957 (Tü 4000)
YP_001828351 (NBRC 13350)
ZP_07296488 (ATCC 53653)
ZP_07293917 (ATCC 53653)
ZP_05522837 (TK 24)
ABY50951 (ATCC 27952)
ZP_06913794 (ATCC 25486)
YP_003487693 (87.22)
ZP_07276967 (AA4)
ZP_06273105 (ACTE)
ZP_07289987 (C)
ZP_05001875 (Mg1)
ZP_07976693 (SA3_actG)
ZP_07273435 (SPB78)
ZP_08453284 (Tü 6071)
ZP_06920565 (ATCC 29083)
CCA53556 (ATCC 10712)
ZP_07608000 (Tü 4113)
ZP_07307300 (DSM 40736)
YP_003335875 (DSM 43021)
YP_003382082 (DSM 17836)
YP_003509930 (DSM 44728)
YP_003116895 (DSM 44928)
Highly conserved residues are shown in red, deviations from otherwise highly conserved residues are shown in dark red. The strictly conserved motifs are shown in boxes. Added nucleotides of corrected sequences are shown in bold.
D A K A T M R T A V S R M L D A W L W E L Q N Q Q L N R V P D P V D Y V E M R R D T F G S D L T K S L A R F A H - - - G D R V P P
E N R R A V R R A V E S M T S S W L W E L A N Q A Q N R I P D P I D Y V E M R R R T F G S D L T M S L S R F S H - - - G R S V P P
E N R R A V R R A V E S M T S S W L W E L A N Q A Q N R I P D P I D Y V E M R R R T F G S D L T M S L S R F S H - - - G R S V P P
T A R T E F R R A I Q D M T D S W V W E L A N Q T Q N R I P D P I D Y I E M R R K T F G S D L T M S L S R L S Q - - - G G E I P M
T A R T E F R R A I Q D M T D S W V W E L A N Q T Q N R I P D P I D Y I E M R R K T F G S D L T M S L S R L S Q - - - G G E I P M
A A R A T F R R T I E V M I D S W L W E L A N Q A H N R I P D P V D Y L E M R R A T F G S D L T M S L C R L A R - - - W H S V P A
G A R E T F R G T V E V M I D S W L W E L A N Q A Q N R I P D P I D Y I E M R R A T F G S D L T M S L A R L A R L A Q E Q T V P P
D S R R T F R R T V C I M L D S W Q W E L A N Q A Q N R I P D P V D Y I E M R R R T F G S D L T M S L S R L G H - - - G R S V P P
S A R R S F R A A I D L M L D S W L W E L A N Q A E N R I P D P I D Y L E M R R A T F G S D L T M A L A R V S G - - - E R L V P T
A A R V T L R K A I I D M T E S W L W E L A N Q I Q N R I P D P V D Y I E M R R K T F G A D L T K G L S R M A L - - - D A T L P E
G A R A K F R Q A V G D M L D S W L W E L A N T A Q N R I P D P V D Y I E M R R A T F G S D L T M S L S R L G R - - - G D T I P A
A S R S L F R K A I L D M T E S W V W E L A N Q I Q N R I P D P I D Y V E M R R Q T F G S D L T M S L S R L A H - - - G D A L P P
N A R T Q F R R A I Q D M T D S W V W E L A N Q I Q N R I P D P I D Y V E M R R K T F G S D L T M S L S R L A Q - - - G S E I P Q
N A R T Q F R R A I Q D M T D S W V W E L A N Q I Q N R I P D P I D Y I E M R R K T F G S D L T M S L S R L S Q - - - G S E I P M
T A R S E F R R A I Q D M T D S W V W E L A N Q T Q N R I P D P I D Y I E M R R K T F G S D L T M S L S R L S Q - - - G G E I P M
A W R Q K F A N Y M Q S Y I E A E W W E I S N V V Q D R V P D P V D Y V E M R R R T A A G D I T I A L A Q Y G L - - - S E I I T P
E T R R R F R A A V D T M I D S W L W E L H N Q H L N R I P D P V D Y F E M R R R T F G S D L T I S L A K F S H - - - G E A V P P
D S R R Q F R K A V E D M T S S W L W E L A N Q T Q N R I P D P V D Y I E M R R R T F G S D M T M S L S R L A N - - - A A V V P A
R L R R R F P E H V R N F V G S W L W E L D N L I Q N R V S D P V D Y L R M R R R T G G S A F R G A L A R H T L - - - G A G L A P
S S R Q S L R R S V E T M T G S W L W E L L N Q T Q N R I P D P V D Y V E M R R K T F G A D L T M S L S K L T L - - - G D V V P A
N A R R L F R K A I Q D M T A S W L W E L A N Q I Q N R I P D P V D Y V E M R R K T F G S D L T M S L S R L A H - - - G D A I P Q
N A R R L F R K A I Q D M T A S W L W E L A N Q I Q N R I P D P V D Y V E M R R K T F G S D L T M S L S R L A H - - - G D A I P Q
K A R R E F R Q T I E V M L D S W V W E L D N Q A I G R I P D P I D Y M E M R R A T F G S D L T M S L C R L R H - - - G N I V P E
D E R R T F R A S V D V M T E S W V W E L S N Q L Q H R I P D P I D Y L E M R R A T F G A D L T L S L C R V G H - - - G P K V P P
E A R R A F R S T V E D M T G S W L W E L A G E M E R R V P D P V D Y I E M R R K T F G A D L T M A L S R L S H - - - G R T L P A
E N R Q A L R T S V E V M T E S W L W E L A N Q A Q N R I P D P V D Y I E M R R R T F G A D L T M A L C R I Q H - - - G R R V P P
S G R R A F R T A V E S M T E S W L W E L A N Q A Q N R I P D P V D Y V E M R R A T F G S D L T M S L C R L G H - - - G R K V P D
A R R R Q F V A A L T E M L E S W L W E L H N R V Q H R V P D P V D Y L E M R R A T F G A D L T M L F G R L R R - - - D T T I P A
E E R R P L K K A V D D M T E A W L W E L S N Q I Q N R V P D P V D Y L E M R R A T F G S D L T L G L C R A G H - - - G P A V P P
A A R R M F R A A V E S M T A S W L W E L A N Q A Q N R I P D P V D Y M E M R R A T F G S D L T M S L C R L G H - - - G R K V P D
E A R G T L K N A V N V M T E S W L W E L A N Q I H H R V P D P V D Y L E M R R Q T F G S D L T L S L C R M G H - - - G P A V P P
E Q R R T M R R S V D V M T A S W V W E L S N Q I Q H R V P D P V D Y L E M R R D T F G S D L T K N L C R M G H - - - A P N V P A
D E R R P L R S A V D T M T E S W V W E L S N Q I Q N R V P D P V D Y L E M R R A T F G S D L T L G L C R A G H - - - G P A V P P
S G R R A F R T A V E S M T E S W L W E L A N Q A Q N R I P D P V D Y V E M R R A T F G S D L T M S L C R L G H - - - G R K V P D
G A R R V F R Q A V D D M L D S W L W E L A N Q A Q H R I P D P V D Y V E M R R H T F G S G L T M S L C R L A Y - - - G R R V P P
Q A R R T F R T A I E D M T S S W L W E L A N Q A H H R I P D P V D Y I E M R R M T F G S D L T M S L C R L A H - - - G Q Q V P P
E E R R P L K K A V D D M T E A W L W E L S N Q I Q N R V P D P V D Y L E M R R A T F G S D L T L G L C R A G H - - - G P A V P P
G A R S S L R S A I D V M L D S W L W E L H N Q A Q H R V P D P V D Y I E M R R L T F G S D L T M S L C R L R H - - - E G E L P P
E A R G S L R A A V D V M L D S W L W E L H N Q A Q H R V P D P V D Y I E M R R F T F G S D M T M S L C R L R H - - - E G E L P A
D E R R G L K E S V D K M T E S W V W E V F N Q I H H R V P D P V D Y L E M R R A T F G S D L T L S M C R M G H - - - G P Q I P P
A E R E A F R A T V E S M I D S W V W E V A N Q A Q N R I P D P I D Y V E M R R R T F G S E L V T S I S R L T N - - - S R T V P P
S A R R T F R E A V E S M T A S W L W E L A N Q A Q N R I P D P V D Y V E M R R A T F G S D L T M S L C R L G H - - - G R K V P E
A G R A E F R A T M V E M L E S W V W E V E N Q F H R R V P D P V D Y A E M R R L T F G S R M T M Y M C R L G Q - - E G R G I P D
R V R A E F R A T L V S M L E S W L W E V D N Q I Q N R I P D P V D Y A E M R R R T F G S H L T M Y L C R L G Q - - Q G R G I P E
V T R A A F R D G V E H M L E S W L W E L G N Q A Q H R V P D P V D Y L E M R R H T F G S E L T M S L S R M R H - - - A A A L P A
V T R A A F R D G V E H M L E S W L W E L G N Q A Q H R V P D P V D Y L E M R R H T F G S E L T M S L S R M R H - - - A A A L P A
V T R A A F R D G V E H M L E S W L W E L G N Q A Q H R V P D P V D Y L E M R R H T F G S E L T M S L S R M R H - - - A A A L P A
D Q R R T L K D S V N V M T E S W L W E L A N Q I Q H R V P D P V D Y L E M R R A T F G S D L T M S M C R M G H - - - G P A V P P
E A R A Q L R Y A L D V M L D S W M W E L H N Q A Q H R V P D P V D Y I E M R R S T F G S E L T M L M C R L R Q - - - D D V L P P
S A R R D L R Q S I E D M T A S W L W E L A N Q T K N R I P D P V D Y I E M R R H T F G S D L T M S L C R L A R - - - G R R V S P
E Q R R P L K D A I D S M T E S W L W E L S N Q L Q N R V P D P V D Y L E M R R A T F G S D L T L S L A R M G H - - - G P A V P P
P A R Q Q F R E A V E D M T A G W L W E L V N Q T Q H R V P D P V D Y I E M R R K T F G S D M T M S L A R L A H - - - S D M M P A
D A R R A F K D A V E T M I D S W L W E L A N Q F Q H R I P D P V D Y L E M R R R T F G S D L T M S L C R I G H - - - G R T V P P
A G R R E F R A A V T V M L D S W L W E L H H Q I Q N R I P D P I D Y I E M R R E T F G A D M T M S L A R L R N - - - G R Q V P R
A G R E G L R R G T Q K M L D S W L W E L E N K A V H R V P D P I D Y V E M R R D A F G S D L T M A L S R I R H - - - - D T I P H

21
 Figure 1. Alignment of geosmin synthases.

721 731 741 751 761 771 781

YP_003098781 (DSM 43827)
AEK39836 (S699)
YP_003763541 (U32)
AEA03338 (CHAB 1432)
AEA03341 (CHAB 2155)
YP_716636 (ACN14a)
YP_483306 (Ccl3)
YP_001509819 (EAN1pec)
YP_004017381 (Eul1c)
YP_003265710 (DSM 14365)
BAJ30389 (KM-6054)
YP_634376 (DK 1622)
YP_001866236 (PCC 73102)
ZP_07114089 (PCC 6056)
ABU93239 (P2r)
ABU93238 (P2r)
YP_001105388 (NRRL 2338)
YP_001107098 (NRRL 2338)
YP_001106173 (NRRL 2338)
YP_001612078 (So ce 56)
ZP_01460669 (DW 4/3-1)
YP_003950745 (DW 4/3-1)
ZP_04705018 (J1074)
NP_823339 (MA-4680)
ADI05189 (BCW-1)
CCB75658 (NRRL 8057)
ZP_08240572 (XylebKG-1)
ZP_06769636 (ATCC 27064)
NP_630182 (A3(2))
ADW07414 (ATCC 33331)
ZP_04685148 (ATCC 14762)
ZP_08289513 (M045)
ZP_07309957 (Tü 4000)
YP_001828351 (NBRC 13350)
ZP_07296488 (ATCC 53653)
ZP_07293917 (ATCC 53653)
ZP_05522837 (TK 24)
ABY50951 (ATCC 27952)
ZP_06913794 (ATCC 25486)
YP_003487693 (87.22)
ZP_07276967 (AA4)
ZP_06273105 (ACTE)
ZP_07289987 (C)
ZP_05001875 (Mg1)
ZP_07976693 (SA3_actG)
ZP_07273435 (SPB78)
ZP_08453284 (Tü 6071)
ZP_06920565 (ATCC 29083)
CCA53556 (ATCC 10712)
ZP_07608000 (Tü 4113)
ZP_07307300 (DSM 40736)
YP_003335875 (DSM 43021)
YP_003382082 (DSM 17836)
YP_003509930 (DSM 44728)
YP_003116895 (DSM 44928)
Highly conserved residues are shown in red, deviations from otherwise highly conserved residues are shown in dark red. The strictly conserved motifs are shown in boxes. Added nucleotides of corrected sequences are shown in bold.
E V Y R T R T L Q N L E N S A A D Y G W M V N D L Y S H R K E V Q Y E G E V H N L V V V V Q N F L D C D L D R A F E V V E R L Q E
A L Y R T R P V Q A I E H S A T D V A C L I N D L Y S Y R K E I Q Y E G E L H N A V L V V R N F L D C T E E R A F G V V T D L V H
A L Y R T R P V Q A I E H S A T D V A C L I N D L Y S Y R K E I Q Y E G E L H N A V L V V R N F L D C T E E R A F G V V T D L V H
Q I Y Y S R P M R S L E N A A A D F A C F T N D I F S Y Q K E I E F E G E I H N C V L V V Q N F L N C D L L K A V E I V N N L M T
Q I Y Y S R P M R S L E N A A A D F A C F T N D I F S Y Q K E I E F E G E I H N C V L V V Q N F L N C D L L K A V E I V N N L M T
E V F G T R P L R A L E N A A A D Y A C L L N D I F S Y Q K E I Q F E G E I H N C V L V V E N F L D C D R G R A V E V V N A L M T
E I Y R T R P I Q A L E N A A A D Y A C L L N D V F S Y Q K E I Q F E G E I H N C V L V V E N F L D C D R E R A L A V V N D L M T
E I Y G T R P I R A L E N S A A D Y S C L L N D I F S Y Q K E I Q F E G E I H N C V L V F Q N F L G C G A E R A I G V V N D L M T
E I Y A T R P I R A L E S S A S D Y A T L L N D L F S Y Q K E I Q F E G E L H N G V L V A E K F L G C S R E R A V T V V N D L M T
A I F R T R P I V N L E N A V A D Y A G I T N D V F S Y Q K E I E F E G E I H N C V L V V E R F L D L D S A S A V A V V N E L M T
E V Y A S G T L R A I E N S A A D Y G C L V N D L H S Y Q K E V E Y E G E F H N L V R V V E N F F S C D Y P V A A D I V A D L M A
E V F H T R P I R S L E N S A A D Y A C L I N D V F S Y Q K E I E F E G E L N N G V L V V Q R F L D L D P A R A V S V V N D L M T
E I Y R T R T M R S L D N S A A D F A C L T N D I F S Y Q K E I E F E G E I H N C V L V V Q N F L N C D L P Q A V E V V N N L M T
E I Y Y S R S M R S L E N S A A D F A C F T N D I F S Y Q K E I E F E G E I H N I V L V V Q N F L N C D L P Q A V E I V N N L M T
E I Y Y S R P M R S L E N A A A D F A C F T N D I F S Y Q K E I E F E G E I H N C V L V V Q N F L N I D I P Q A V E I V N N L M T
E I N S N R Q I Q S L N N I T I D W I G L S N D I V S Y R K E M E F E G G I H N S V L V F K R F L D C P L Q E S V N I V N N L L T
E I Y R T R T I R N M E N S A I D Y A T M L N D V F S Y R K E I E Y E G E V H N A V L V V R N F L D C D Q D R A F E I V G D L M T
E I Y R T R V M R E L E W S A Q D Y A C F T N D L F S Y Q K E I E F E G E V H N M V L V V E N F L G V D R L T A R D V V A D L M K
A V F D T P E M R A L H E N W A D V G P L R N D L F S Y H K E V D R E T E V T N G V L A V Q R F F D C G L Q Q A A A V V A D L A E
P I F R T R T M R G L D N T T A D V G G W I N D L Y S Y R K E I E F E G E L S N F V L V A E R F L E C D A Q R A K E V V N D L I T
E I F H T R P V R G L E N S A A D Y A C L T N D I F S Y Q K E I E Y E G E L N N G V L V V Q R F L E I E P P Q A V E I V N D L M T
E I F H T R P V R G L E N S A A D Y A C L T N D I F S Y Q K E I E Y E G E L N N G V L V V Q R F L E I E P P Q A V E I V N D L M T
E V Y R S G T I Q A M E K A V S D Y G A L I N D V F S Y Q K E V E F E G E I H N A V L V T Q N F F D C D Y P T G L A Y V D A L M N
E I Y R S G P V R S L E N A A V D Y G M L I N D V F S Y Q K E I E Y E G E V H N A I L V V Q N F F G C D Y P T A L G V I N D L M T
E V L R A R P V Q A M E H A V G D F A G L Q N D L H S F R K E V E F S G E I H N G V L V V Q N F F G C G T S E A L H I V A D L V N
E V Y A S G P I R S L E N S A V D Y A C L L N D V F S Y Q K E I E F E G E V H N G I L V V Q N F F N C D Y P T G L G I V H D L M T
E V Y R S G P M R S L E N A A A D Y A C L M N D L F S Y Q K E I E Y E G E V H N G V L V V Q N F F G V D Y P T G V A I V H D L M N
E V F A S G T L R T L E R S A Q D W C T L L N D I H S Y R K E I E T E G E P N N A V L V V R T F F G C D D A S A V D V V H D L M R
E V Y R S G P V R S L E N A A I D Y A C L L N D V F S Y Q K E I E Y E G E I H N A V L V V Q N F F G V D Y P A A L G V V Q D L M N
E V Y R S G P L R S L E N A A A D Y A C L M N D L F S Y Q K E I E Y E G E V H N G V L V V Q N F F G V D Y P T G V A I V H D L M K
E V Y R S G P V R S L E N A A I D Y A C L L N D V F S Y Q K E I E Y E G E I H N A I L V V Q T F F G C D Y P R A L G I V H D L M D
E V Y T S G P I C S L E N A A M D Y A V L L N D V F S Y Q K E I E Y E G E I H N A I L V V Q N F F Q C G Y A E A V R V V G D L M N
E V Y R S G P V R S L E N A A I D Y A C L L N D V F S Y Q K E I E F E G E M H N A V L V V Q N F F G V D Y P T A L P V V Q D L M N
E V Y R S G P M R S L E N A A A D Y A C L M N D L F S Y Q K E I E Y E G E V H N G V L V V Q N F F G V D Y P T G V A I V H D L M N
D I Y D S G P V R S L E S A A M D Y A T L L N D V F S Y Q K E I E F E G E V H N G V L V V Q N F F D C D Y P T G V A I V Q D L M T
E I Y R S G P L R S L E N A A A D Y A C L L N D V F S Y Q K E I E Y E G E V H N G V L V V Q N F F N C D Y P T A L G I V H D L M T
E V Y R S G P V R S L E N A A I D Y A C L L N D V F S Y Q K E I E Y E G E I H N A V L V V Q N F F G V D Y P A A L G V V Q D L M N
E L Y A S G P V R G L E N A A M D Y A C L I N D L F S Y Q K E I E Y E G E V H N A V L V V Q T F F D C D R P T A A A M T D A L M R
E L Y A S G P V R A L E N A A M D Y A C L L N D L F S Y Q K E V E Y E G E I H N A V L V V Q T F F D C D Y P T G F A M T A D L M H
E V Y R S G P V R S L E N A A I D Y G C L I N D V F S Y Q K E I E Y E G E V H N A I L V V Q N F F G C D Y P A A L G V V H D L M T
E I H R N R V I E A L E D S A A D Y A W L I N D L F S Y R K E I E Y E G A L H N A V L V V R S F L D V D Q D R A I Q I V A D L A E
A V Y R S G P L R S L E N A A A D Y A C L L N D L F S Y Q K E I E Y E G E V H N G V L V V Q N F F G V D Y P T G V A I V H D L M N
E L Y R S G T I R S L E N A A A D A G C L V N D I F S Y R K E V E F E G E V H N H V L V T R N F F D I G Y P E A L R I C H S L L V
E I Y A S G T I R S L E N A A A D A A C M I N D I F S Y Q K E V E V E G E V H N F V L V T R N F F D I G Y P E A L R I C H D L L T
G V L D S G T V R A L E N A V A D Y G C L I N D V F S Y Q K E V Q Y E G E L H N L V L V V E N F F G C D Y P T A F R M V E H L M A
G V L D S G T V R A L E N A V A D Y G C L I N D V F S Y Q K E V Q Y E G E L H N L V L V V E N F F G C D Y P T A F R M V E H L M A
G V L D S G T V R A L E N A V A D Y G C L I N D V F S Y Q K E V Q Y E G E L H N L V L V V E N F F G C D Y P T A F R M V E H L M A
E V Y R S G P V R S L E N A A V D Y A C L L N D V F S Y Q K E I E Y E G E V H N S I L V V Q N F F G I D Y P T A L R V V H D L M T
E L Y R S G T V R A L E N A V M D Y G A L I N D L F S Y Q K E I E V E G E V H N G V L V L Q T F F D C D Y P T A V A M V D D L M R
Q I Y R S G P V R S L E N A A M D Y A T L L N D V F S Y Q K E I E F E G E V H N G V L V V Q N F F D C D Y P T G V A V V N D L M T
E L Y R S G P V R S L E N A A F D F A C L L N D V F S Y Q K E I E Y E G E I H N A I L V V Q S F F G C D Y P A A L G V V H D L M S
E I Y Q T R V M R E L D T A A Q D Y A C F T N D L F S Y Q K E I E F E G E V H N L V L V V E N F L E V D R W K A R D V V A D L M T
E I Y R T R P I R A L E S A A A D Y A C L L N D V F S Y R K E I Q Y E G E L H N G V L V V R N F L D C D L S T A F A V V N E L M T
A V Y D S R P I R E L E N S A V D Y A C L L N D L F S Y Q K E I E F E G E V I N G V L V V Q E F L G C S V P E A M A I V N D L M T
E V L R S R P V L A L E H A A A D W G C L V N D V F S Y Q K E I Q F D G D V H N A V L V V Q D F F D C D R D R A L E I V S S L M A

22
 Figure 1. Alignment of geosmin synthases.

791 801 811 821 831 841

YP_003098781 (DSM 43827)
AEK39836 (S699)
YP_003763541 (U32)
AEA03338 (CHAB 1432)
AEA03341 (CHAB 2155)
YP_716636 (ACN14a)
YP_483306 (Ccl3)
YP_001509819 (EAN1pec)
YP_004017381 (Eul1c)
YP_003265710 (DSM 14365)
BAJ30389 (KM-6054)
YP_634376 (DK 1622)
YP_001866236 (PCC 73102)
ZP_07114089 (PCC 6056)
ABU93239 (P2r)
ABU93238 (P2r)
YP_001105388 (NRRL 2338)
YP_001107098 (NRRL 2338)
YP_001106173 (NRRL 2338)
YP_001612078 (So ce 56)
ZP_01460669 (DW 4/3-1)
YP_003950745 (DW 4/3-1)
ZP_04705018 (J1074)
NP_823339 (MA-4680)
ADI05189 (BCW-1)
CCB75658 (NRRL 8057)
ZP_08240572 (XylebKG-1)
ZP_06769636 (ATCC 27064)
NP_630182 (A3(2))
ADW07414 (ATCC 33331)
ZP_04685148 (ATCC 14762)
ZP_08289513 (M045)
ZP_07309957 (Tü 4000)
YP_001828351 (NBRC 13350)
ZP_07296488 (ATCC 53653)
ZP_07293917 (ATCC 53653)
ZP_05522837 (TK 24)
ABY50951 (ATCC 27952)
ZP_06913794 (ATCC 25486)
YP_003487693 (87.22)
ZP_07276967 (AA4)
ZP_06273105 (ACTE)
ZP_07289987 (C)
ZP_05001875 (Mg1)
ZP_07976693 (SA3_actG)
ZP_07273435 (SPB78)
ZP_08453284 (Tü 6071)
ZP_06920565 (ATCC 29083)
CCA53556 (ATCC 10712)
ZP_07608000 (Tü 4113)
ZP_07307300 (DSM 40736)
YP_003335875 (DSM 43021)
YP_003382082 (DSM 17836)
YP_003509930 (DSM 44728)
YP_003116895 (DSM 44928)
Highly conserved residues are shown in red, deviations from otherwise highly conserved residues are shown in dark red. The strictly conserved motifs are shown in boxes. Added nucleotides of corrected sequences are shown in bold.
A R V R Q F Q H V L D V E M P Q M F E D L G L D D V A R R A L L D Y G D E L K D W M A G I V N W H E N C V R Y H D E D L - - - - -
A R L A E F E H A A A I E L P A L V R D H A I E P A V E E M L L G Y V Q E L R D W L A G I L N W H E N V G R Y T E P E L - - - - -
A R L A E F E H A A A I E L P A L V R D H A I E P A V E E M L L G Y V Q E L R D W L A G I L N W H E N V G R Y T E P E L - - - - -
A R A Q Q F Q H I V E T E L P A L F D D F N L D K N T R E K L L K Y I E K L E Q W M C G V L K W H T K V D R Y K E F E L - - - - -
A R A Q Q F Q H I V E T E L P A L F D D F N L D K N T R E K L L K Y I E K L E Q W M C G V L K W H T K V D R Y K E F E L - - - - -
A R M R Q F E H V V D R E L P D L F D R L D L D G E A R A A I V S Y A R E L Q N W L A G I L R W H Q G T H R Y E E A E L - - - - -
S R I R Q F E H I V A H E L P A L F D S F A L D A S A R Q A L L G Y A R E L Q N W L A G I L R W H E G T H R Y E E S E L - - - - -
A R L R E F E H V V D V E L P A L F D T Y E L T E E A R D V L R G Y V G E L K S W L A G V L R W H Q G T R R Y D E A E L - - - - -
A R M L Q F E H I V E Q E L P A L F E T Y D L A E D A Q G A L L T Y V E D L R N W L A G I L R W H Q G T R R Y D E S E L - - - - -
G R M R E I E H I V A V E L P V L V R E Y E L D S A A E A D L Y R Y V H R L Q Q Y T A G V L R W H F A V D R Y K E P E L - - - - -
S R L S Q F R H V V D S E L P L L K Q D F A L D A E A A R A L D G Y V R E L E D W M A A V L N W H Q Q C D R Y T E D V L - - - - -
A R M Q Q F E Y I I A N E L E P L A R N F N L D G K A Q D K L K Q Y V Q K L Q W W M S G V L I W H Q T V D R Y K E F E L - - - - -
S R A L Q F Q L I V A T E L P V L F D D F D L D A S T R E K L L G Y V K K L E Q W M C G V L K W H I T V D R Y K E F E L - - - - -
A R A Q Q Y Q H I V D T E L P A L L D D F N L D E S T R E K L L G Y V K K L E N W M C G V L K W H I T V D R Y K E F E L - - - - -
A R A Q Q F Q H I V K T E L P A L F E D F N L D E S T R E K L M K Y V E K L E Q W M C G V L K W H T K V D R Y K E F E L - - - - -
A R L H E F E H I V A - E L P T L F E Q L N L D Q N T R Q Q C F E Y V K R L E I W I A G N F V W T N K T R R Y N D F P V - - - - -
A R M K Q F Q Y T V D D E L P V L C E D F G L S S E S R A V L T R Y A D E L R D W M S G I L N W H R E C V R Y K D E D L - - - - -
A R M R Q F E R I L A E E L P T L I D E F E L D E A A R T A L T R Q C D E L K D W T S G I L E W H R R C V R Y T D A E L R R T R S
V R L R R F T A V A E Q E L P A L A H R F E P G R A P R E E L D R Y V R G L H D W L A G E L A W S Q V T G R Y R E P S V - - - - -
A R V L E F E H I V A T E L P R I L D E F E L D K A A R E A V L G Y V E K Q R L Y M A G V L H W H V V T S R Y V D A E L - - - - -
A R M R Q F E H T V K M E L P L L I R S T G L D A K A Q E K L R T Y V E K L Q R W M C G V L R W H M T V D R Y K E F E L R N T R K
A R M R Q F E H T V K M E L P L L I R S T G L D A K A Q E K L R T Y V E K L Q R W M C G V L R W H M T V D R Y K E F E L - - - - -
A R L E Q F E H I V A H E L P V L Y E D F Q L D G A A R K V L D G Y V R Q L E N W L S A I L T W H R G C K R Y K E E D L - - - - -
Q R M H Q F E H V A A H E L P L L Y K D F K L P Q E V R D I M D G Y V V E L Q N W M S G I L K W H Q D C H R Y G A A D L - - - A R
S R M R E F E R I V S T E L P A L V E D L A L D A T V R A T L D G Y T R E L Q N Y M S G V L V W H R Q T P R Y A E S E L - - - - -
S R M K Q F Q H V A A H E L P V L Y D D L K L T A D V R R I L D G Y V R E L Q N W M A G I L T W H Q G C H R Y Q E S E L R Y R P G
S R L R Q F L H V A E V E L P V L C D D F G L D A E A R E T L S G Y V R E L E H W I A G I L I W H R G C R R Y R E E D L - - - - -
G R L R Q F L H V K E D E L P V L Y E D L A L S G T E R A A V G R Y V G E L E D F Q A A M F N W H R T V R R Y G A E D P - - - - -
Q R M R Q F E H V V A H E L P V V Y D D F Q L S E E A R T V M R G Y V T D L Q N W M A G I L N W H R N V P R Y K A E Y L - - - A G
G R M E Q F Q H V A E H E L P L L C E E F G L D T E A R E T L S G Y V R E L R H W L A G I L I W H R G C R R Y R E E D L - - - - -
R R L S Q F E H V V A H E L P I L Y E D F G L S G E A R A I M G E Y V A D L R N W L A G I L N W H R S V D R Y K A E W L - - - A G
Q R M E Q F E H V S A H E L P I L Y E D F A L S P E A R A T V E G Y V V Q L Q N W M S G I L N W H R N V D R Y R A E Y L - - - A R
Q R M R Q F E H V T E H E L P V V Y D D F Q L S E D A R A I M S G Y V E D L R N W M A G I L N W H R N V D R Y K D E Y L - - - S G
S R L R Q F L H V A E V E L P V L C D D F G L D A E A R E T L S G Y V R E L E H W I A G I L I W H R G C R R Y R E E D L - - - - -
S R M R Q F Q H V A A H E F P V L Y E D F G L S A E G R A V L E G Y V R D L Q N W M A G I L T W H R E V A R Y R E E E L - - - - R
S R M R Q F Q H V A R H E L P V V Y D D F K L G S D A R R I L D G Y V R E L E N W L S G I L T W H Q G C R R Y K E D D L - - - - -
Q R M R Q F E H V V A H E L P V V Y D D F Q L S E E A R T V M R G Y V A D L Q N W M A G I L N W H R N V P R Y K A E Y L - - - A G
S R L E Q F L H T K E H E L P L V C E E F G L D E G G S A A L G T Y V R E L E D W L A G I L N W H R K V R R Y K E E D L - - - - -
S R L E Q F L H V K E H E L P V V C D E F G L G E N G R A A L A T Y V R E L E D W L A G I L N W H R R V R R Y K E E D L - - - - -
Q R M R Q F E H V V A H E L P V V Y D D F R L S R E A R D I M G G Y V T D L Q N W M A G I L N W H R N V D R Y K P E F L - - - A R
A R L T E F R H A V E V G L P G L F A D Y G L D E E A R N T L T D Y A D L L K N L M T G I A N W H Q K S A R Y T D E E L - - - - D
G R M R Q F Q H V A E H E L P V L Y D D F G L D A E A R E I L A G Y V V E L Q H W L A G I L I W H R D C R R Y R E E D L - - - - -
R R V E E F Q H V A A E Q L P V L C D D W R L D A G A R A G L D A Y V G G L R D W L A G V L H W H R T T R R Y R D E D L - - - - -
R R T E E F E H I V A N Q L P L L Y D D W K L D A G A R A G L D A Y V G E L E D W L A G I L N W H R K V R R Y R E E D L - - - - -
A R L D Q F E H A V A N E L P V L Y E D F G L T S G Q I E T M G T Y L G E L R D W L A G I L N W H R G V R R Y D A A F L P G C L P
A R L D Q F E H A V A N E L P V L Y E D F G L T S G Q I E T M G T Y L G E L R D W L A G I L N W H R G V R R Y D A A F L - - - - =
A R L D Q F E H A V A N E L P V L Y E D F G L S H E Q I G T M E T Y L G E L R D W L A G I L N W H R G V R R Y D A A F L - - - - =
Q R M Q Q F E H V A A H E L P V V Y D D F A L S E K A R E V M R G Y V T D L Q H W M A G I L H W H R T V D R Y K A D H L - - - A R
G R R R Q Y E H L K A R E V P L M Y E E F G L D A A G R Q A F E A Y L R E L E D W L A G I L N W H R K V R R Y G A E D V - - - - -
S R M R Q F Q H V A E H E F P V L Y D D F G L A P E A R K T M D G Y V R E L R H W M T G I M N W H R E V P R Y R E A E L L R A H R
Q R M R Q F E H V V E H E L P V L Y D D F Q L T D E G R A V M G E Y V A D L R N W L A G I L N W H Q S V D R Y K D E W L - - - S R
A R M Q Q F E H I V A N G L P A L F D D F A L D E Q A R R I L T R H A D D L K E W M S G I L E W H R R C A R Y T E A E L - - - - -
A R M R Q F E Q L V A V D L P V L F D D Y E L G D D V R R V L T G Y A E E L Q N W M S G I L V W H A G C R R Y D E G A Q L - - - -
A R M R Q F Q N I E A N E L P I V F E E F G L A E E A R A A V G K Y V D E L K D W M A A I L N W H R G I T R Y E E S E L - - - - -
A R L A Q F H H S A E V E I P A L A D T M R L S S A T R E A L D L H I Q E L R D W L T G V L N W H Q R T G R Y A E A E Q H R L L V

23
 Figure 1. Alignment of geosmin synthases.

851 861 871 881 891 901 911

YP_003098781 (DSM 43827)
AEK39836 (S699)
YP_003763541 (U32)
AEA03338 (CHAB 1432)
AEA03341 (CHAB 2155)
YP_716636 (ACN14a)
YP_483306 (Ccl3)
YP_001509819 (EAN1pec)
YP_004017381 (Eul1c)
YP_003265710 (DSM 14365)
BAJ30389 (KM-6054)
YP_634376 (DK 1622)
YP_001866236 (PCC 73102)
ZP_07114089 (PCC 6056)
ABU93239 (P2r)
ABU93238 (P2r)
YP_001105388 (NRRL 2338)
YP_001107098 (NRRL 2338)
YP_001106173 (NRRL 2338)
YP_001612078 (So ce 56)
ZP_01460669 (DW 4/3-1)
YP_003950745 (DW 4/3-1)
ZP_04705018 (J1074)
NP_823339 (MA-4680)
ADI05189 (BCW-1)
CCB75658 (NRRL 8057)
ZP_08240572 (XylebKG-1)
ZP_06769636 (ATCC 27064)
NP_630182 (A3(2))
ADW07414 (ATCC 33331)
ZP_04685148 (ATCC 14762)
ZP_08289513 (M045)
ZP_07309957 (Tü 4000)
YP_001828351 (NBRC 13350)
ZP_07296488 (ATCC 53653)
ZP_07293917 (ATCC 53653)
ZP_05522837 (TK 24)
ABY50951 (ATCC 27952)
ZP_06913794 (ATCC 25486)
YP_003487693 (87.22)
ZP_07276967 (AA4)
ZP_06273105 (ACTE)
ZP_07289987 (C)
ZP_05001875 (Mg1)
ZP_07976693 (SA3_actG)
ZP_07273435 (SPB78)
ZP_08453284 (Tü 6071)
ZP_06920565 (ATCC 29083)
CCA53556 (ATCC 10712)
ZP_07608000 (Tü 4113)
ZP_07307300 (DSM 40736)
YP_003335875 (DSM 43021)
YP_003382082 (DSM 17836)
YP_003509930 (DSM 44728)
YP_003116895 (DSM 44928)
Highly conserved residues are shown in red, deviations from otherwise highly conserved residues are shown in dark red. The strictly conserved motifs are shown in boxes. Added nucleotides of corrected sequences are shown in bold.
V R H F G T - - - - - - - - A G S P V A G P A V G A A G P G P G A L P A L G G P S G L G T S A A R V L A K A R - - - - - - - - - -
R Y H P A A - - - - - - - A T A T T F G G P T - G I G T S S L R I S S L L P A G H - - - - - - - - - - - - - - - - - - - - - - - -
R Y H P A A - - - - - - - A T A T T F G G P T - G I G T S S L R I S S L L P A G H - - - - - - - - - - - - - - - - - - - - - - - -
- R N S A S - - - - - - - P I V R L L N G P T - G F G T S A A H I R S L V G A T N S V I - - - - - - - - - - - - - - - - - - - - -
- R N S A S - - - - - - - P I V R L L N G P T - G F G T S A A H I R S L V G A T N S V I - - - - - - - - - - - - - - - - - - - - -
R Y H P A A - - - - - - - - D R R P F G S P T - G L G T S A A D V R R L A S R - - - - - - - - - - - - - - - - - - - - - - - - - -
R Y H P A A - - - - - - - - G V R P F G G P T - G L G T S S A H V R P R P A A A A G A A G D S E M - - - - - - - - - - - - - - - -
R H H P A V - - - - - - - - G V R P F G G P V - G L G T S A A D I R R A L S G K S G Q P T A L T G S - - - - - - - - - - - - - - -
R Q H P A A - - - - - - - - G V P P F G S P V - G L G T S A A R L A S L T R R S S G G L G A T S S P S G P R G H R P P P A R P R -
Q A E R A R - - - - - - - - - K H A I G R P S - G L G T S A S R I A A L F A A G G H A T R - - - - - - - - - - - - - - - - - - - -
R R H F A - - - - - - - - - - - - P P T A V T P E L G M S A A R I L Q T I G R - - - - - - - - - - - - - - - - - - - - - - - - - -
R A S R K L - - - - - - - - A P R L S S G P T - G L G T S A A R I T S L F A N L R S G A - - - - - - - - - - - - - - - - - - - - -
- R N S L - - - - - - - - - A G R L L S G P R - G L G T S A R R I G S L I G Q G S L K S L L G Q - - - - - - - - - - - - - - - - -
- R N S A S - - - - - - - P I V R L L N G P T - G F G T S A A R L T S L V G A S S F S G K M P L N N S D R F V G K - - - - - - - -
- R D S A S - - - - - - - - P V V R A I D T S K G F D T S T L Q I G S F I S K G N H F V K N G - - - - - - - - - - - - - - - - - -
- - P N L P - - - K - - - - - A E P V V S K T P F L G N S A F K I G S L V G T S N P F V N Q R - - - - - - - - - - - - - - - - - -
- R H D A V - - - - - S Q G L A A L L R G P S - G L G T S A V E L R - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
E H H H G T G P - E P H L P L R R R L S G P T - G I G T S A A R L A R R G S S A T G L N R - - - - - - - - - - - - - - - - - - - -
S A V G A D - - - - - - - - L P A A P L G I T G A A G - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
E R Q P I - - - - - - - - - - E R A I D G A R - G F G A S A A R I S S F F G A G R A G A G A P A A K A D L S M G T A R S V P V K Q
P R R G G W - - - E - - - - D P R D G A P P R P A S R R S L G A T G A E V E K K L E K S G S S T - - - - - - - - - - - - - - - - -
- R N T R K - - - - - - - - P R R V V G G P S - G R G T S A A R L A S L F G G N R A E V E K K L E K S G S S T - - - - - - - - - -
S H H S P G - - - V - - - - - R R V I E A A P R G I G P N G L G A S S L G T A A A R V F A G G - - - - - - - - - - - - - - - - - -
R A H G F V - - - - - - - - P D R A P S A P F T A W A A P V A R - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
R R P V G L - - - - - - - - P P R T S Y G P T - G L G T A A A R L A A T L G G G R A A A I G - - - - - - - - - - - - - - - - - - -
P R T G V S A P - G - - - - V G A V P A G P T - G L G T S A A R L P L L T S G R P R - - - - - - - - - - - - - - - - - - - - - - -
- R G G A G - - - - - - - - A P W R P G G P T - G L G T S A A Q M P R V M A R L A G S P G - - - - - - - - - - - - - - - - - - - -
- - - - A - - - - - - - - - P G S P L L G P P T G L G T S A A R L A S A L A R - - - - - - - - - - - - - - - - - - - - - - - - - -
R T H G F L - - - - - - - - P D R I P A P P V P R S S P A L T H - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
- R R G S G - - - - - - - - A P W H L G G P T - G L G T S A A Q V I R L L T E Q G R F S P A G R V R R A A V - - - - - - - - - - -
R V H G F L - - - - - - - - P G R P P A L P V L V - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
R S H G F L - - - - - - - - P D R G P A L P V - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
R V H G F L - - - - - - - - P H R T P A P P V L V - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
- R G G A G - - - - - - - - A P W R P G G P T - G L G T S A A Q V P R V M A R L A G S A G - - - - - - - - - - - - - - - - - - - -
Y Q A G A P - - - R - - - - L L R Q L F G P T - G L G T S A A R L P V G A G R - - - - - - - - - - - - - - - - - - - - - - - - - -
R R D L P G - - - R - - - - P H R R L D I P T - G L G T S A A H L P R L T G R R - - - - - - - - - - - - - - - - - - - - - - - - -
R T H G F L - - - - - - - - P D R I P A P P V P R S S P A L T H - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
- R G G A V - - - - - - - - - P R R L G A P T - G L G T S A A R L S L P S R L S G V G V - - - - - - - - - - - - - - - - - - - - -
- R G G A A - - - - - - - - - P W R L G A P T - G L G T S A A R I S L P A Q R S A I R V - - - - - - - - - - - - - - - - - - - - -
R A H N F V - - - - - - - - P D R P P T L S L T P L R T G - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
R V R N T Q - - - L - - - - - H T A A A A P I - - D T A K A L Q H P G C T G T P R G P K P L L T S H S Q - - - - - - - - - - - - -
- R R G T G - - - T - - - - - P W H L S G P T - G F G T S A A Q V T R L L T Q G R F S P A G T A R P A A V - - - - - - - - - - - -
- R P P V D - - - A - - - - - - L S A L V L S S G F G M A A A R I P G R - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
- - H G L P - - - D - - - - - L L S S G V W S S S F G M S A A R L S L P R - - - - - - - - - - - - - - - - - - - - - - - - - - - -
Y G A G P E - - - T - - - - A V R G A G V P G P G A G A E G A F A T R P A A V G A A A A V A V P G A A L A - - - - - - - - - - D -
- - - P G C L - - - P - - - - Y G A G P E T A V R G A G V P G A A G A G A T G P G A G A G G V F A M R P A A V G A - - - - A A V A
- - - P G C - - - - - - - - - - - - L P Y G S V P E T A V R G A G V P G A A G P G P G A G G V F A T G P A A V G A - - - - A A V A
R T H G F L - - - - - - - - P D R P P A L P M A G - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
- R S G S G - - - S - - - - - - T L L R G P T - G L G T S T A R V A A L L G R - - - - - - - - - - - - - - - - - - - - - - - - - -
R P V G A G A - - G S G A R P V A P W H G P T - G L G T S A A R I P V P A G A R S - - - - - - - - - - - - - - - - - - - - - - - -
R A H R F L - - - - - - - - P D R P P A V P V P A L G - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
R R S R L P - - - G - - - - A P A G F S L L P A G L G T S A V R V G A G R R G - - - - - - - - - - - - - - - - - - - - - - - - - -
- - - - - - - - - - - - - H L P R K L L G G A T G L G T S A E S I G R S L A M K R L E G A L - - - - - - - - - - - - - - - - - - -
R R - - - - - - - - - - - - P T T A L G G P M - G L G T N G N R P P R P H V P V V A P N P G A D R P S S A V A A M N T P V N H P K
T R L G H R S P W R P K T P S S V P S G A P S G I S G F S G P P F P A H T T S P T A A T P - - - - - - - - - - - - - - - - - - - -

24
 Figure 1. Alignment of geosmin synthases.

921 931 941 951 961 971

YP_003098781 (DSM 43827)
AEK39836 (S699)
YP_003763541 (U32)
AEA03338 (CHAB 1432)
AEA03341 (CHAB 2155)
YP_716636 (ACN14a)
YP_483306 (Ccl3)
YP_001509819 (EAN1pec)
YP_004017381 (Eul1c)
YP_003265710 (DSM 14365)
BAJ30389 (KM-6054)
YP_634376 (DK 1622)
YP_001866236 (PCC 73102)
ZP_07114089 (PCC 6056)
ABU93239 (P2r)
ABU93238 (P2r)
YP_001105388 (NRRL 2338)
YP_001107098 (NRRL 2338)
YP_001106173 (NRRL 2338)
YP_001612078 (So ce 56)
ZP_01460669 (DW 4/3-1)
YP_003950745 (DW 4/3-1)
ZP_04705018 (J1074)
NP_823339 (MA-4680)
ADI05189 (BCW-1)
CCB75658 (NRRL 8057)
ZP_08240572 (XylebKG-1)
ZP_06769636 (ATCC 27064)
NP_630182 (A3(2))
ADW07414 (ATCC 33331)
ZP_04685148 (ATCC 14762)
ZP_08289513 (M045)
ZP_07309957 (Tü 4000)
YP_001828351 (NBRC 13350)
ZP_07296488 (ATCC 53653)
ZP_07293917 (ATCC 53653)
ZP_05522837 (TK 24)
ABY50951 (ATCC 27952)
ZP_06913794 (ATCC 25486)
YP_003487693 (87.22)
ZP_07276967 (AA4)
ZP_06273105 (ACTE)
ZP_07289987 (C)
ZP_05001875 (Mg1)
ZP_07976693 (SA3_actG)
ZP_07273435 (SPB78)
ZP_08453284 (Tü 6071)
ZP_06920565 (ATCC 29083)
CCA53556 (ATCC 10712)
ZP_07608000 (Tü 4113)
ZP_07307300 (DSM 40736)
YP_003335875 (DSM 43021)
YP_003382082 (DSM 17836)
YP_003509930 (DSM 44728)
YP_003116895 (DSM 44928)
Highly conserved residues are shown in red, deviations from otherwise highly conserved residues are shown in dark red. The strictly conserved motifs are shown in boxes. Added nucleotides of corrected sequences are shown in bold.
- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
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G R S - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
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- - - - - - - V Q M T A P A G V P A A G S V P R Q A A S G A A P A T A S G A A P E T E A S A V P A V A A T A P G W Q Y P S G I G T
V P G A A L A D V Q M T A P A G V P A A G S V P R Q A A S G S A P A T A S G A T P E T D A S A V P A V A A T A P R W Q Y P S G I G
V P G A A L A D V Q M T A P A G V P A A G S V P R Q A A S G A A P A T A S G A A P E T D A S A V P S V A A T A P G W Q H P S G I G
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P A I A T I G V G A V K H - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
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25

YP_003098781 (DSM 43827)
AEK39836 (S699)
YP_003763541 (U32)
AEA03338 (CHAB 1432)
AEA03341 (CHAB 2155)
YP_716636 (ACN14a)
YP_483306 (Ccl3)
YP_001509819 (EAN1pec)
YP_004017381 (Eul1c)
YP_003265710 (DSM 14365)
BAJ30389 (KM-6054)
YP_634376 (DK 1622)
YP_001866236 (PCC 73102)
ZP_07114089 (PCC 6056)
ABU93239 (P2r)
ABU93238 (P2r)
YP_001105388 (NRRL 2338)
YP_001107098 (NRRL 2338)
YP_001106173 (NRRL 2338)
YP_001612078 (So ce 56)
ZP_01460669 (DW 4/3-1)
YP_003950745 (DW 4/3-1)
ZP_04705018 (J1074)
NP_823339 (MA-4680)
ADI05189 (BCW-1)
CCB75658 (NRRL 8057)
ZP_08240572 (XylebKG-1)
ZP_06769636 (ATCC 27064)
NP_630182 (A3(2))
ADW07414 (ATCC 33331)
ZP_04685148 (ATCC 14762)
ZP_08289513 (M045)
ZP_07309957 (Tü 4000)
YP_001828351 (NBRC 13350)
ZP_07296488 (ATCC 53653)
ZP_07293917 (ATCC 53653)
ZP_05522837 (TK 24)
ABY50951 (ATCC 27952)
ZP_06913794 (ATCC 25486)
YP_003487693 (87.22)
ZP_07276967 (AA4)
ZP_06273105 (ACTE)
ZP_07289987 (C)
ZP_05001875 (Mg1)
ZP_07976693 (SA3_actG)
ZP_07273435 (SPB78)
ZP_08453284 (Tü 6071)
ZP_06920565 (ATCC 29083)
CCA53556 (ATCC 10712)
ZP_07608000 (Tü 4113)
ZP_07307300 (DSM 40736)
YP_003335875 (DSM 43021)
YP_003382082 (DSM 17836)
YP_003509930 (DSM 44728)
YP_003116895 (DSM 44928)
Highly conserved residues are shown in red, deviations from otherwise highly conserved residues are shown in dark red. The strictly conserved motifs are shown in boxes. Added nucleotides of corrected sequences are shown in bold.
Figure 1. Alignment of geosmin synthases.
981 991
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S A A R V G P G A V R -
T S A A R V G P G A V R
T S A A R V G P G A V R
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26
A) Nostoc punctiforme SAG 60.79 (GQ287652)
Chitinophaga pinensis DSM 2588 (NC_013132)
B) 100 ZP_05522837 (Streptomyces lividans TK 24)
48 NP_630182 (Streptomyces coelicolor A3(2))
91 Ktedonobacter racemifer DSM 44963 (NZ_ADVG01000001) ZP_07309957 (Streptomyces griseoflavus Tü 4000)
99 Herpetosiphon aurantiacus DSM 785 (CP000875) 38 66
ZP_07307300 (Streptomyces viridochromogenes DSM 40736)
Roseiflexus castenholzii DSM 13941 (CP000804) 33
ZP_04685148 (Streptomyces ghanaensis ATCC 14762)
100 Haliangium ochraceum DSM 14365 (NC_013440) 43 ZP_06920565 (Streptomyces sviceus ATCC 29083)
96 Plesiocystis pacifica SIR-1 (NR_024795) ZP_08289513 (Streptomyces griseoaurantiacus M045)
15
96 Sorangium cellulosum So ce 56 (NC_010162) 90
NP_823339 (Streptomyces avermitilis MA-4680)
100 Myxococcus xanthus ATCC 25232 (DQ768116)
Stigmatella aurantiaca ATCC 25190 (DQ768127) YP_003487693 (Streptomyces scabiei 87.22)
15
Rubrobacter xylanophilus DSM 9941 (NC_008148) CCA53556 (Streptomyces venezuelae ATCC 10712)
24
ZP_06769636 (Streptomyces clavuligerus ATCC 27064)
Catenulispora acidiphila DSM 44928 (NC_013131) 51 21 ZP_07273435 (Streptomyces sp. SPB78)
Actinosynnema mirum DSM 43827 (NC_013093) 100
82 ZP_08453284 (Streptomyces sp. Tü 6071)
92 Saccharopolyspora erythraea NRRL 2338 (NC_009142) 75
99 ZP_07976693 (Streptomyces sp. SA3_actG)
Amycolatopsis mediterranei U32 (NC_014318)

Kribbella flavida DSM 17836 (NC_013729) ZP_06913794 (Streptomyces pristinaespiralis ATCC 25486)
93 24 100
100 Micromonospora olivasterospora DSM 43868 (X92613) ABY50951 (Streptomyces peucetius subsp. caesius ATCC 27952)
Micromonospora aurantiaca ATCC 27029 (NC_014391)
Micromonospora sp. L5 (NC_014815) ZP_04705018 (Streptomyces albus J1074)
100 Frankia sp. Eul1c (NC_014666) ZP_07296488 (Streptomyces hygroscopicus ATCC 53653)
Frankia sp. EAN1pec (NC_009921) 99
89 63 ZP_07608000 (Streptomyces violaceusniger Tü 4113)
Frankia sp. Ccl3 (NC_007777) 13
100 Frankia alni ACN14a (NC_008278) CCB75658 (Streptomyces cattleya NRRL 8057)
75
77 ZP_07293917 (Streptomyces hygroscopicus ATCC 53653)
100 60 27
Stackebrandtia nassauensis DSM 44728 (NC_013947) BAJ30389 (Kitasatospora setae KM-6054 (NBRC 14216))
Thermomonospora kurvata DSM 43183 (NC_013510)
70 ZP_06273105 (Streptomyces sp. ACTE)
68 Nocardiopsis dassonvillei DSM 43111 (X97886) 56
Streptosporangium roseum DSM 43021 (NC_013595) ADW07414 (Streptomyces flavogriseus ATCC 33331)
99
76 Planobispora rosea JCM 3166 (AB028654) 47 YP_001828351 (Streptomyces griseus subsp. griseus NBRC 13350)
100 ZP_08240572 (Streptomyces cf. griseus XylebKG-1)
Streptomyces cattleya NRRL 8057 (FQ859185)
Streptomyces albus subsp. albus DSM 40313 (AJ621602) ZP_05001875 (Streptomyces sp. Mg1)
„Streptomyces bingchenggensis“ BCW-1 (CP002047) 99
61 Streptomyces violaceusniger Tü 4113 (NZ_AEDI01000202.1) 24 ZP_07289987 (Streptomyces sp. C)

100 Streptomyces hygroscopicus subsp. hygroscopicus NBRC 13472 (AB184428) YP_003116895 (Catenulispora acidiphila DSM 44928)
48
Streptomyces ghanaensis ATCC 14672 (NZ_DS999641) ADI05189 (“Streptomyces bingchenggensis“ BCW-1)
Streptomyces griseoaurantiacus NBRC 15440 (AB184676)
Streptomyces pseudogriseolus NRRL B-3288T (DQ442541) YP_001612078 (Sorangium cellulosum So ce 56)
Streptomyces griseoflavus LMG 19344 (AJ781322) 15 23 YP_003509930 (Stackebrandtia nassauensis DSM 44728)
Streptomyces ambofaciens NBRC 12836 (AB184182) 92
YP_003335875 (Streptosporangium roseum DSM 43021)
Streptomyces viridochromogenes NRRL B-1511T (DQ442555)
Streptomyces sp. SA3_actG (NZ_ADXA01000222) YP_003382082 (Kribbella flavida DSM 17836)
Streptomyces coelicolor A3(2) (NC_003888)
92 YP_004017381 (Frankia sp. EuI1c)
Streptomyces odorifer DSM 40347 (Z76682) 43
77 YP_001509819 (Frankia sp. EAN1pec)
Streptomyces arenae ISP 5293 (AJ399485)
70 68
YP_716636 (Frankia alni ACN14a)
Kitasatospora setae KM-6054 (U93332) 92
YP_483306 (Frankia sp. Ccl3)
Streptomyces scabiei 87.22 (NC_013929)
Streptomyces clavuligerus ATCC 27064 (NZ_CM000913) 78
100Streptomyces griseus subsp. griseus NBRC 13350 (NC_010572) 46 YP_001105388 (Saccharopolyspora erythraea NRRL 2338)
95
YP_003098781 (Actinosynnema mirum DSM 43827)
Streptomyces cf. griseus XylebKG-1 (NZ_ADFC00000000.2) 49
Streptomyces peucetius NBRC 100596 (AB249907) ZP_07276967 (Streptomyces sp. AA4)
Streptomyces sp. ACTE (HM235473) 100 YP_003763541 (Amycolatopsis mediterranei U32)
Streptomyces flavogriseus CBS 101.34T (AJ494864) 100 AEK39836 (Amycolatopsis mediterranei S699)
Streptomyces filamentosus NBRC 12767 (AB184130)
98
Streptomyces pristinaespiralis ATCC 25486 (ABJI00000000) YP_003265710 (Haliangium ochraceum DSM 14365)
93 Streptomyces venezuelae ATCC 10712 (FR845719) ZP_01460669 (Stigmatella aurantiaca DW 4/3-1)
Streptomyces avermitilis MA-4680 (NC_003155) 39 YP_003950745 (Stigmatella aurantiaca DW 4/3-1)
99 100
Streptomyces sviceus ATCC 29083 (AB184559) 82
YP_634376 (Myxococcus xanthus DK 1622)
Streptomyces lasaliensis JCM 3373 (HQ537060) 0.1
Streptomyces griseocarneus JCM 5010 (AY999766) 84 ZP_07114089 (Oscillatoria sp. PCC 6056)
93
YP_001866236 (Nostoc punctiforme PCC 73102)
100 ABU93238 (Phormidium sp. P2r)
0.1 ABU93239 (Phormidium sp. P2r)
58
66 AEA03341 (Anabaena ucrainica CHAB 2155)
100 AEA03338 (Anabaena ucrainica CHAB 1432)

YP_001107098 (Saccharopolyspora erythraea NRRL 2338)

Figure 2. Comparison of a 16S rRNA gene-based phylogenetic tree (A) with the phylogenetic YP_001106173 (Saccharopolyspora erythraea NRRL 2338)
tree based on the amino acid sequences of geosmin synthases (B). Groups of related strains
and their geosmin synthases are shown in the same colour. Outgroup 27
Figure 3. Total ion chromatograms of representative headspace extracts from bacteria (part I).

28
Figure 3. Total ion chromatograms of representative headspace extracts from bacteria (part II).

29
Figure 3. Total ion chromatograms of representative headspace extracts from bacteria (part III).

30
Figure 3. Total ion chromatograms of representative headspace extracts from bacteria (part IV).

31
Figure 3. Total ion chromatograms of representative headspace extracts from bacteria (part V).

32
Figure 3. Total ion chromatograms of representative headspace extracts from bacteria (part VI).

33
Figure 3. Total ion chromatograms of representative headspace extracts from bacteria (part VII).

34
Figure 3. Total ion chromatograms of representative headspace extracts from bacteria (part VIII).

35
Figure 3. Total ion chromatograms of representative headspace extracts from bacteria (part IX).

36
Figure 3. Total ion chromatograms of representative headspace extracts from bacteria (part X).

37
Figure 3. Total ion chromatograms of representative headspace extracts from bacteria (part XI).

38
Figure 3. Total ion chromatograms of representative headspace extracts from bacteria (part XII).

39
Figure 3. Total ion chromatograms of representative headspace extracts from bacteria (part XIII).

40
Figure 3. Total ion chromatograms of representative headspace extracts from bacteria (part XIV).

41
Figure 3. Total ion chromatograms of representative headspace extracts from bacteria (part XV).

42
Figure 3. Total ion chromatograms of representative headspace extracts from bacteria (part XVI).

43
Figure 4. Mass spectrum of an unidentified compound (A) and a similar mass spectrum of the known compound rosifoliol (B). Due to the similarities between the
mass spectra the unknown compound is suggested to be a stereoisomer of rosifoliol. Biosynthetic considerations lead to the suggested structure of 8-epi-rosifoliol
(Scheme 1 of main text).

44
Figure 5. Alignment of 2-MIB synthases. Highly conserved amino acid residues are shown in red. Highly conserved motifs are shown in boxes. Missing parts of sequences are indicated by question marks.
Position 1 11 21 31 41 51 61
YP_003765432 (U32) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
YP_003115314 (DSM 44928) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
BAJ32779 (KM-6054) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - M P S D P S D P P F L V A A A A A V A E A V A A T
BAK26793 (NRRL 8178) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ZP_04609212 (ATCC 39149) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
YP_003680543 (DSM 43111) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - M P P L P R M S A S A A S P E V A A L V D R L L R D S R R P G R P D L L A S P A
YP_347573 (Pf0-1) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
YP_001105919 (NRRL 2338) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
YP_003510780 (DSM 44728) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
CAJ89344 (ATCC 23877) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - M P D S G P L G P H S P D H R P T P A T T V P D A P
ADI12075 (BCW-1) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ZP_08234965 (XylebKG-1) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - M P V P E L P P P R S S L P E A V T R F G A S V L G A V A A R A H D
NP_733742 (A3(2)) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - M P D S G T L G T P P P E Q G P T P P T T L P D V P
ZP_05801975 (ATCC 33331) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - M T A P E L P P P R S S L P E A A S R F G A H L L A S A A A H A L D
YP_001822781 (NBRC 13350) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - M P V P E L P P P R S S L P E A V T R F G A S V L G A V A A R A H D
ZP_05520932 (ATCC 53653) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
BAI77523 (NRRL 3382) - - - - M P D S G S L G P P T S L P E Q P P A P P A T A P D A P A A T V T D R P V T S S V A H F L A G L H P P V T R P S S P P S P S M P P A
YP_003486275 (87.22) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - M P T P A S S A A V P D A A A P A A A P D H R P V
ZP_07279119 (AA4) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ZP_07290591 (C) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ZP_05002186 (Mg1) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
CCA60397 (ATCC 10712) M P E P G P S P L Q S S L P S A A A H F G A H V L A R A A A P A G A P A P V P T D A A P T A R A V P A A P S G P V V P S G P V V P A A L A V
ZP_07604855 (Tü 4113) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - M P A S E P S P P Q P S P P G A G S P
ZP_07301463 (DSM 40736) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ZP_05521425/6 (TK 24) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - M P D S G T L G T P P P E Q G P T P P T T L P D V P

45
Figure 5. Alignment of 2-MIB synthases. Highly conserved amino acid residues are shown in red. Highly conserved motifs are shown in boxes. Missing parts of sequences are indicated by question marks.
71 81 91 101 121 131 141
YP_003765432 (U32) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - M T G E S T K T T C P Q W S A G E A A G L - - - - - - L A G P F G L G T S A A
YP_003115314 (DSM 44928) - - - - - - - - - - - M T L I A D T G R S A A A V L P G L A A L T G P P P A R P P W E A A A P E A R S E H W - - - - H S S D V D P G P E S P
BAJ32779 (KM-6054) A A A A A A A P A T A T A T A T T P A T V E T V P A P S V S G A S V P A P S V P E P V - A V P E A D G L Q R I - - - L R G P S G L G T G G L
BAK26793 (NRRL 8178) - - - - - - - - - - M S A A D A L S G F A A D A L S G F A A D A L S G F A A A I L A A T G R A P S A E L S Q V - - - A A G P T A L D R L T D
ZP_04609212 (ATCC 39149) - - - - - - - - - - - M F L P S L D S G S A G K L T G L A A T I L S D M V G A P P A E L T R L T G P T L R - - - - - L N G P T G L G T A A L
YP_003680543 (DSM 43111) A P R T P P A P R L P V G S R Q V G T S A L R L R L P D S H V A D Q S P G A L A V R T G L R L P G H P G G L G - - - V P G P T A P A P G R P
YP_347573 (Pf0-1) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
YP_001105919 (NRRL 2338) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - M I E L I G H E T P V P
YP_003510780 (DSM 44728) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - M T T Q A R V L P - - T G P V G L G T D L A
CAJ89344 (ATCC 23877) A S K P P D V A V T P T A S E F L A A L H P P V P I P S P S P P S G S A S A A A D T P D A T T V G S A L Q R I - - - L R G P T G P G T A A L
ADI12075 (BCW-1) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ZP_08234965 (XylebKG-1) S E A T V G G P S G G R P L P S P P A - - - G L S F G P P S P A A P P A D V P A P E A P G R - - G A D L E R L - - - L C G P H G L G T A G L
NP_733742 (A3(2)) A P V I P S A S V T S A A S D F L A A L H P P V T V P D P A P P P P P A P A A G N P P D T V T G D S V L Q R I - - - L R G P T G P G T T S L
ZP_05801975 (ATCC 33331) V E G V T G G P P G T T S L P A P P V T S A S A L A A R P L P A A P A A G A P A P S - - - - - - - A G L D R I - - - L R G P S G L G T A G L
YP_001822781 (NBRC 13350) S E A T V G G P S G G R P L P S P P A - - - G L S F G P P S P A A P S A D V P A P E A P G R - - G A D L E R L - - - L C G P H G L G T A G L
ZP_05520932 (ATCC 53653) M S A P E F S P P P S L S A P P P V D I A P A D T A A V D I A P A D T T P A D T A A A E V R S A S V G L E R I - - - L R G P S G L G T A S L
BAI77523 (NRRL 3382) S S N P S S P P S S S M P P A S W A P P S P L S P P A P S L P P T S P P A T A P E T S A A T G S D S V V R R V - - P - V G P T G L G T T A L
YP_003486275 (87.22) A P T A S G F L A A L H P P V A L L V P P P T V P T P T P S P P V E P S A A V P G P P D T T A A D D A L R R I - - - L R A P T G P G T A S L
ZP_07279119 (AA4) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - M T G I A A V R P D L A C P S S L - - W - A G P V G L G T S A A
ZP_07290591 (C) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
ZP_05002186 (Mg1) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
CCA60397 (ATCC 10712) P S A P A A P F A P S A P V V P V A P V G P L V R V A S A A P S A P A A S A A P A A V G D S P D R P D L G L V - - - L R G P T G L G T A G L
ZP_07604855 (Tü 4113) S G A F V L A E A A A R A R D I R A A F G G G R S L A P S L P A P P P A D A - - P A T G A R T A G A D L G R I - - - L R G P S G L G T V G L
ZP_07301463 (DSM 40736) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - M E R S T S V N T P L T P P A P S P
ZP_05521425/6 (TK 24) A P V I P S A S V T S A A S D F L A A L H P P V T V P D P A P P P P P A P A A G N P P D T V T G D S V L Q R I - - - L R G P T G P G T T S L

46
Figure 5. Alignment of 2-MIB synthases. Highly conserved amino acid residues are shown in red. Highly conserved motifs are shown in boxes. Missing parts of sequences are indicated by question marks.
151 161 171 181 191 201 211
YP_003765432 (U32) R I A T R F A P S P G H D P A Y A Y L - - - - P - - - - - - W G D G S A A P L Y C P V Q G R V D D G L A A E V D R R L V A W A - - D G C G F
YP_003115314 (DSM 44928) A R P P Q H P E R L A P A P I E L R A - - - - - - - - - - - W G D G S H S P L Y C P A P S R L D E A L A A D V N A R L V A W A - E R I G L H
BAJ32779 (KM-6054) F R V S P L P E R L P E P G A E V P G A P E A S A S L P E P V E G V P V P G L Y F H P V A E P D P V R V A E V S R R I K D W A V D E V D L F
BAK26793 (NRRL 8178) S T G L G R S A F R I P R S P M L P - - - - - - - - - - - P P T D D G V P E L F C P G P V R D D P A L G E T V N D G I V E W A - G Q V G I Y
ZP_04609212 (ATCC 39149) R I S P P S A P A A V P P A P D - - - - - - - - - - - - - - - - - R G R P E L F C P G P V R D D P A L G E E V N D R V V E W A - E Q V G I Y
YP_003680543 (DSM 43111) G T G V P P E G P V V A S E P E G P E - - - - - - - - - - - V D G E R I P A L Y C P P A V R D D P A L G D E V D E R L L V W A - E E M G V Y
YP_347573 (Pf0-1) - - - - - M N Q S S S A R T P R S A - - - - - - - - - - - - T A P F I V R A V R C P P P T R I D E A L G Q E V N E R L M E W I - S N I G I F
YP_001105919 (NRRL 2338) S Q Q Q H T G G V R G T S A C T P P - - - - - - - - - - - - G V G E R T T V L Y C P P P P P E R P E V A A E I N R R V V V W M - Q G L G L G
YP_003510780 (DSM 44728) R L F S V P P P K A A G S A G K A S P T - - P - - - - - - - K G S V T V P E L Y C P D A V R D D A A L G A E V D R R L A V W G R E E I G L -
CAJ89344 (ATCC 23877) A L S V R H D P P S L P G S P A P A E - - - - P - - - - - - A A G R A V P G L Y H H P V P E P D P A R V E E V S R R I K R W A E D E V Q L Y
ADI12075 (BCW-1) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - M R V E E V G R R I K A W A L D E V D L Y
ZP_08234965 (XylebKG-1) R L T P G K E R P V P A T A R E - - - - - - - - - - - - - - - - G R P I P G L Y H H P V P E P D E A R V E E V S R R I K A W A L D E V S L Y
NP_733742 (A3(2)) A P A V R Y G R Q P G P E A P A S A P - - - - P - - - - - - A A G R A V P G L Y H H P V P E P D P V R V E E V S R R I K R W A E D E V Q L Y
ZP_05801975 (ATCC 33331) R L F P R D E P Q A P P S P P A A A - - - - - - - - - - - - A E G R P V P G L Y H H T V T E P D P G L V E E V S R R I K A W A L D E V S L Y
YP_001822781 (NBRC 13350) R L T P G K E R P V P A T A R E - - - - - - - - - - - - - - - - G R P I P G L Y H H P V P E P D E A R V E E V S R R I K A W A L D E V S L Y
ZP_05520932 (ATCC 53653) H P A A R R E E P R T P P T P P E A P - - - - - - - - - - - A E G S P I P G L Y H H P V P E P D P G R V E E V S R R I K A W A L D E V D L Y
BAI77523 (NRRL 3382) S L A R R Q A A V P P D A V P A P S G P - - - S - - - - - - A E G P V V P G L Y H H P I P E P D P V R V A E V S R R I K R W A E D E V R L Y
YP_003486275 (87.22) V V A D R F A P P L P S P V S R A P V E - - - P - - - - - - A A G R A V P G L Y H H P V P E P D P V R V E E V S R R I K R W A E E E V Q L Y
ZP_07279119 (AA4) R I A A Q F A P S G G Q D P A Y A E R E - - - W - - - - - - - G D G S A S P L Y C P V V R R V D E P L A E E V D R R L V A W A E D - - C G F
ZP_07290591 (C) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - M A E L S R R I K A W A L D E I Q L Y
ZP_05002186 (Mg1) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - M P E P D P V R V E E V S R R I K E W A V D E V E L Y
CCA60397 (ATCC 10712) S L A R R E A V P E L P E P P A P E A P - - - - - - - - - - P A G R P I P G L Y F H P I A E P D P V R V D E V S R R I K A W A L D E I Q L Y
ZP_07604855 (Tü 4113) H L A R C E G A S A P V E Q V E P P T A - - - P - - - - - - A E G T P V P G L Y H H P V P E P D P V R V A E V S R R I K T W A L D E V D L Y
ZP_07301463 (DSM 40736) G F T P P S P P S G P P S V T R G G H P - - - - - R - - - - - P G E P V P G L R H R P A V P P D P E K V E E I D R R L E A W A - H E L K L F
ZP_05521425/6 (TK 24) A P A V R Y G R Q P G ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? I K R W A E D E V Q L Y

47
Figure 5. Alignment of 2-MIB synthases. Highly conserved amino acid residues are shown in red. Highly conserved motifs are shown in boxes. Missing parts of sequences are indicated by question marks.
221 231 241 251 261 271 281
YP_003765432 (U32) T G K G L E Q I A G A G F G R L A M L A H T D C D D P D R L L V A A Q L N A V W W A A D D Y Y A D D S T L - G A A P T E L P P R L A L V M A
YP_003115314 (DSM 44928) A G H - L E E F A K T G F G R L I T L A H P E C D D P D L L L V S A Q M N A A W W A S D D Y Y A D E T D L - G A V A E A L P E R L A L V S S
BAJ32779 (KM-6054) P D E W E G Q F D G F S V G R Y M V V C H P D A P T V E H V M V A T R L M V A E N A V D D C Y C E D H - - - G G S P V G L G S R L L L A H T
BAK26793 (NRRL 8178) P G Q - L D R L R A Y N F G R L I M L T H P A T N D P D R L L A A A K C V V A E W A T D D Y V V D E V S L - G A D P A V V G S R L A K L H A
ZP_04609212 (ATCC 39149) P G R - L D R L R A G N F G R L I M L A H P A T S D P D R L L A A T K C V V A E W A A D D Y M V D E V S L - G A D P T V V G S R L A K L H V
YP_003680543 (DSM 43111) P G Q - L D R V R S A G F G R L I M L A H P E T D D P D R L L A A A K C A L A E W S V D D H Y V D G E A E - E A Q P E L L G Q R L A I A H S
YP_347573 (Pf0-1) A G K - E E K I R A S D F G R Y A M L C H A D T N N P D R L L L V A Q C F A A L F A V D D H Y C D D Q S L - G G R P E T V A E S L S F A L T
YP_001105919 (NRRL 2338) G E D N V A G V Y K H D P G R G I T L C H P G S Q D V E R M T A A G K M I V A E T A V D D Y F C E T N S R R D A N D Q T I G P N L S L A Q S
YP_003510780 (DSM 44728) P D E H V D K L G K C G Y G R L I M L T H P D C D D P D R L L A A A K C A V A E W S V D D L Y L D G D S A - E P E P E R L G P R L A L A Y A
CAJ89344 (ATCC 23877) P E D W E G E F D G F S V G R Y M V A C H P D A P T V D H L M L A T R L M V A E N A V D D C Y C E D H - - - G G S P V G L G G R L L L A H T
ADI12075 (BCW-1) P E D W E D Q F D G F S V G R Y M V A C H P D A P T V D H L M L A T R L M V A E N A V D D C Y C E D H - - - G G S P I G L G G R L L L A H T
ZP_08234965 (XylebKG-1) P E E W E E Q F D G F S V G R Y M V G C H P D A P T V D H L M L A T R L M V A E N A V D D C Y C E D H - - - G G S P V G L G E R L L L A H T
NP_733742 (A3(2)) P E E W E G Q F D G F S V G R Y M V G C H P D A P T V D H L M L A T R L M V A E N A V D D C Y C E D H - - - G G S P V G L G G R L L L A H T
ZP_05801975 (ATCC 33331) P E D W E E Q F D G F S V G R Y M V G C H P D A P T V D H L M L A T R L M V A E N A V D D C Y C E D H - - - G G S P V G L G E R L L L A H T
YP_001822781 (NBRC 13350) P E E W E E Q F D G F S V G R Y M V G C H P D A P T V D H L M L A T R L M V A E N A V D D C Y C E D H - - - G G S P V G L G E R L L L A H T
ZP_05520932 (ATCC 53653) P E D W E D Q F D G F S V G R Y M V A C H P D A P T V D H L M L A T R M M V A E N A V D D C Y C E D H - - - G G S P I G L G G R L L L A H T
BAI77523 (NRRL 3382) P E E W E G Q F D G F S V G R Y M V A C H P D A P T V D H L M L A T R L M V A E N A V D D C Y C E D H - - - G G S P V G L G G R L L L A H T
YP_003486275 (87.22) P E E W E G Q F D G F S V G R Y M V A C H P D A P T T D H L M L A T R L M V A E N A V D D C Y C E D H - - - G G S P V G L G G R L L L A H T
ZP_07279119 (AA4) D G E G L D Q I A N A G F G K L A M L A H P D S S D P D R L L I A A Q L N A V W W A A D D Y Y A D D S S L - G A S P T E L P P R L A L V M A
ZP_07290591 (C) P D D W E G E F D G F S T G R Y M V A C H P D A P T I E H L M I A A R L M V A E N A V D D M Y C E D H - - - G G S P I G L G G R L L L A H S
ZP_05002186 (Mg1) P P E W E D Q F D G F S V G R Y M V A C H P D A P T V D H L M I A T R L M V A E N V V D D C Y C E D H - - - G G S P V G L G G R L L L A H T
CCA60397 (ATCC 10712) P E D W E G E F D G F S V G R Y M V A C H P D A P T V E H L M L A T R L M V A E N A V D D C Y C E D H - - - G G S P V G L G G R L L L A H T
ZP_07604855 (Tü 4113) P E D W E D Q F D G F S V G R Y M V A C H P D A P T V D H L M L A T R L M V A E N A V D D C Y C E D H - - - G G S P I G L G G R L L L A H T
ZP_07301463 (DSM 40736) P P A W T G D F A G F Q F G R A V V L Q H P G A A D L D R L T V A G E L L L A E N L V D S C Y C S E D E D R G G S S R G L G G R L V I A Q S
ZP_05521425/6 (TK 24) P E E W E G Q F D G F S E G R Y M V G C H P D A P T V D H L M L A T R L M V A E N A V D D C Y C E D H - - - G G S P V G L G G R L L L A H T

48
Figure 5. Alignment of 2-MIB synthases. Highly conserved amino acid residues are shown in red. Highly conserved motifs are shown in boxes. Missing parts of sequences are indicated by question marks.
291 301 311 321 331 341 351
YP_003765432 (U32) A M D P V A - P A G E F S G P L E E A L R R D P I L V G L H S G I D H L G R H G S P V L V Q R V C Y A T F S M F V S W D A Y A A W R H T G R
YP_003115314 (DSM 44928) A L D P P P - P V G E F T P P L Q D A V V S D P V L V S L R S A L A H V T R H A S A A Q V M R V R H T T H Q M Y V S W N A Y N A W R H A G I
BAJ32779 (KM-6054) A L D P V H - T T A E Y A P D W A E S L G S D A P R R A Y R S A M R Y F T E L A S P S Q A D R F R H D M A R L H L G Y L A E A S W S Q D D H
BAK26793 (NRRL 8178) V V D P A R - L P A R Y A P Q L D A Y R R D E P I A T A F R S A M E H L A R Y T T V A Q L G R F Q H Q L G I L F L A W N Q E A D W H V N G R
ZP_04609212 (ATCC 39149) I V D P A R - L P A R Y A P Q L D Q Y R R E E P I A T A F R S A M Q H L S R Y V S V P Q L A R F Q H Q M S I L F V A W N Q E A D W H V N R R
YP_003680543 (DSM 43111) V I D Q A H - L P L R Y A P Q L E E V V R A D P V M R A L R S S L D G L G L Y A T A S Q V R R L R H E L A I M F V A Y N Q E G V W L A T G H
YP_347573 (Pf0-1) A I D P V Y - L P S P F D K E L L K Q Q M C D P V I R G L L A Y M K R V A Q F C T P S Q V A R V R Q I T I A M F V T M A A E G P W R L Y G T
YP_001105919 (NRRL 2338) A I D A P R - L T P D L Q A L W N K C R D D H P V L R A Q H E A F G D L E R I S S P A Q A Q R V R H D I A Q L Y L G Y N A E N G W R L L N R
YP_003510780 (DSM 44728) A M A P A R V P P P P Y R E P V K E R I R E D L C L R M I H S A W K N L A R Y A S H T Q V F R L R H E L A V M F V A Y N Q E A E W H I T E R
CAJ89344 (ATCC 23877) A I D P F H - T T A E Y A P P W R E S L T S D A P R R A Y R S A M D Y F V R A A T P S Q A D R Y R H D M A R L H L G Y L A E A A W A Q T D H
ADI12075 (BCW-1) A L D P L H - T T K E Y Q P P W A E S L H S D A P R R A Y R S A M E Y F L Q A A S A S Q A D R F R H D M A R L H L G Y L A E A A W A Q T D H
ZP_08234965 (XylebKG-1) A L D P L Y - T A R E Y Q P G W A A S L H A D A P R R A Y R S A M D Y F V R A A G P S Q A D R L R H D M A R L H L G Y L A E A A W A Q Q D Q
NP_733742 (A3(2)) A I D H F H - S T A E Y T P T W Q A S L A A D A P R R A Y D S A M G Y F V R A A T P S Q S D R Y R H D M A R L H L G Y L A E G A W A Q T G H
ZP_05801975 (ATCC 33331) A L D P L Y - T T E E Y R P Q W A E S L H A D A P R R A Y R S A M E Y F V R A A S A S Q A D R F R H D M A R L H M G Y L A E A A W A Q L D H
YP_001822781 (NBRC 13350) A L D P L Y - T A R E Y Q P G W A A S L H A D A P R R A Y R S A M D Y F V R A A G P S Q A D R L R H D M A R L H L G Y L A E A A W A Q Q D Q
ZP_05520932 (ATCC 53653) A L D P L Y - T A K E Y Q P Q W A E S L H S D A P R R A Y R S A M E Y F V R E A S A S Q A D R L R H D M A R L H M G Y L A E A A W A Q T D H
BAI77523 (NRRL 3382) A L D H L H - T T A E Y A P E W S E S L G S D A P R R A Y R S A M D H F V R A A T P S Q A D R Y R H D M A R L H L G Y L A E A A W A E T G H
YP_003486275 (87.22) A L D H F H - T T A E Y A P A W Q E S L A S D A P R R A Y R S A M D H F V G A A T P S Q A D R Y R H D M A R L H L G Y L A E A A W A Q T G H
ZP_07279119 (AA4) A M D P V A - P A G E F T A P L E E A L R A D P I R V G F H S A V D H L G R H G S P V L V Q R V C Y S T F A M F V S W D A Y A A W R H T G R
ZP_07290591 (C) A L D P L H - T T E E Y R P R W A E S L H E D A P R R S Y R S A M E Y F V Q Q S T P S Q A D R F R H D M A R L H M G Y L A E A A W A Q T D H
ZP_05002186 (Mg1) A L D A L H - T T R E Y A P D W E E S L H S D A P R R A Y R S A M E Y F T R E A T A S Q A D R Y R H D M A R L H L G Y L A E A A W A Q T D Y
CCA60397 (ATCC 10712) A L D P V H - T T K E Y Q P L W A E S L Y A D A P R R A Y R S A M E Y F V N Q T T A S Q A D R F R H D M A R L H L G Y L A E A A W A E T N H
ZP_07604855 (Tü 4113) A L D P L H - T T K E Y Q P R W A E S L H S D A P R R A Y R S A M E Y F V Q A A G A S Q A D R Y R H D M A R L H M G Y L A E A A W A Q T H H
ZP_07301463 (DSM 40736) A L D P Y H - G P P E V E R E W R R G L T A D G P L R S Y H C A L R D Y A A F A T P S Q T N R F V H D V A R L H L G Y V A E A A W A Q T R Y
ZP_05521425/6 (TK 24) A I D H F H - S T A E Y T P T W Q A S L A A D A P R R A Y D S A M G Y F V R A A T P S Q S D R Y R H D M A R L H L G Y L A E G A W A Q T G H

49
Figure 5. Alignment of 2-MIB synthases. Highly conserved amino acid residues are shown in red. Highly conserved motifs are shown in boxes. Missing parts of sequences are indicated by question marks.
361 371 381 391 401 411 421
YP_003765432 (U32) Y P P A W E Y L A A R Q H D S F Y T S M T L V D A V G G Y E L A A P F Y Y D P R V R E A M M R A G T A S V L V N D L H S V A K D A A D E K P
YP_003115314 (DSM 44928) T P Q A W R Y L A A R Q H D S F Y T S M I L I D V V G G Y E L P P E L F A E P L F H R A L T Q A G T A A V L V N D L A S A A R E - A G E D P
BAJ32779 (KM-6054) V P E V W E Y L A M R Q F N N F R P C P T I T D T V G G Y E L P A D L Q A M P Q T Q R V I A L A G N A T T I V N D L Y S Y T K E L A A P G R
BAK26793 (NRRL 8178) T P P V W E Y L V Q R H L N N F L P P M V L V D A V A G Y E L S P D E F F D P R V R R A Y T T A A L A N V L L N D I H S G T C E - - - S D T
ZP_04609212 (ATCC 39149) T P P V W E Y L V Q R H L N S Y L P P M I L V D A V A G Y E L S P E E F F H P L V R R A F T T A G L A A V L L N D I Y S G A Q E - - - S D T
YP_003680543 (DSM 43111) R P P V W E F L M H R H E N S F V P C M A L I D A V A G Y E L P H Q V F S E P S V R R V F T L A G S A S V I V N D L Y S M G K E - - - D V T
YP_347573 (Pf0-1) Q P T V A E Y L A S R Q V N S F W P C L V L I D L I G G Y E V P A N T Y S R P D I H H V T A L A S L A T T L V N D L Y S A Y K E H L N E T G
YP_001105919 (NRRL 2338) L P P V W Q Y L A N R Q M N S F R P C L N L T D A L D G Y E L A P Q L Y A H P L V Q D C T A R A T L I A T L Y N D L A S C E R E I R E H G L
YP_003510780 (DSM 44728) V P P T W E F L L H R W E N A F C P C M V L T D V V G G Y E V P V F E Y A D P R V R R V F T T A G V A S V L V N D L Y S L E K E K E I N G F
CAJ89344 (ATCC 23877) V P E V W E Y L A M R Q F N N F R P C P T I T D T V G G Y E L P A D L H A R P D M Q R V I A L A G N A T T I V N D L Y S Y T K E L D S P G R
ADI12075 (BCW-1) V P E V W E Y L A M R Q F N N F R P C P T I T D T V G G Y E L P A D L H A Q P A M Q R V I A L A G N A T T I V N D L Y S Y T K E L A S P G R
ZP_08234965 (XylebKG-1) V P E V W E Y L A M R Q F N N F R P C P T I T D T V G G Y E L P A D L H A Q A A M Q K V I A L A S N A T T I V N D L Y S Y T K E L A A P G R
NP_733742 (A3(2)) V P E V W E Y L A M R Q F N N F R P C P T I T D T V G G Y E L P A D L H A R P D M Q R V I A L A G N A T T I V N D L Y S Y T K E L N S P G R
ZP_05801975 (ATCC 33331) V P E V W E Y L A M R Q F N N F R P C P T I T D T V G G Y E L P A D L H A Q A G M Q K V I A L A G N A T T I V N D L Y S Y T K E L D A P G H
YP_001822781 (NBRC 13350) V P E V W E Y L A M R Q F N N F R P C P T I T D T V G G Y E L P A D L H A Q A A M Q K V I A L A S N A T T I V N D L Y S Y T K E L A A P G R
ZP_05520932 (ATCC 53653) V P E V W E Y L V M R Q F N N F R P C P T I T D T V G G Y E L P A D L H A R P E M Q R V I A L A G N A T T I V N D L Y S Y T K E L A S P G K
BAI77523 (NRRL 3382) V P E V C E Y L A M R Q F N N F R P C P T I T D T V G G Y E L P A D L H A R P D M Q R V I A L A G N A T T I V N D L Y S Y T K E L D S P G R
YP_003486275 (87.22) V P E V W E Y L A M R Q F N N F R P C P T I T D T V G G Y E L P A D L H A R P D M Q R V I A L A G N A T T I V N D L Y S Y T K E L D S P G H
ZP_07279119 (AA4) Y P P A W E Y L A A R Q H D S F Y T S M T L I D A V G G Y E L P A P F Y Y D P R V R E A M M R A G T A T V L V N D L H S V A K D A A D E N P
ZP_07290591 (C) M P E V W E Y L V M R Q F N N F R P C P T I T D T V G G Y E L P S D L H A Q P A M Q R V L A L A G N V S T I V N D L Y S Y T K E L A S P G R
ZP_05002186 (Mg1) V P Q V W E Y L A M R Q F N N F R P C P T I T D T V G G Y E L P A D L H A Q A A V Q R V I A L A G N A T T I V N D L Y S Y T K E L A S P G R
CCA60397 (ATCC 10712) V P E V W E Y L S M R Q F N N F R P C P T I T D S V G G Y E L P A D L H A Q P A M Q R V L A L A G N A T T I V N D L Y S Y T K E L A S P G K
ZP_07604855 (Tü 4113) V P E V W E Y L A M R Q F N N F R P C P T I T D T V G G Y E L P A D L H A T P A M Q R V I A L A G N A T T I V N D L Y S Y T K E L A S P G H
ZP_07301463 (DSM 40736) T P R V W E Y L V M R Q F N N F R P C L S I V D A V D G W E L P E A V Y A R P E I Q R I T A L A C N A T T I V N D L Y S F T K E L A G D P D
ZP_05521425/6 (TK 24) V P E V W E Y L A M R Q F N N F R P C P T I T D T V G G Y E L P A D L H A R P D M Q R V I A L A G N A T T I V N D L Y S Y T K E L N S P G R

50
Figure 5. Alignment of 2-MIB synthases. Highly conserved amino acid residues are shown in red. Highly conserved motifs are shown in boxes. Missing parts of sequences are indicated by question marks.
431 441 451 461 471 481 491
YP_003765432 (U32) V C N M V L Q I A A D R D C P V E E A V Q A T V E L H N K I V H E F E A G H R E L M - A V P S P E L Q R F L V G V R S W M G G G F T W H A T
YP_003115314 (DSM 44928) D C N L V L L L A A E R D C S I A E A T E Q V V A L H N D V V R G F E H S R A A L A - A V P S P E L Q R F V L G A R A W M G G C L E W H - D
BAJ32779 (KM-6054) H L N L P V V I S E R E G L S D K D G Y L K A V E V H N E L M R A F E A E S A A L A A A C P A P Q L L R F L R G V A A W V D G N H H W H Q T
BAK26793 (NRRL 8178) D F N L P R V I S I E E G C S L R D A V T R T V E I H N E L M H A F V A D A A T L S - L I G S P N L R R F L A D I W A W L G G S R E W H A T
ZP_04609212 (ATCC 39149) D F N L P R V I A A E H N C A L E E A I T R T V E I H N E L M H E F V A D A V T L S - L T G S P M L R R F L A D T W A W L G G S R E W H A T
YP_003680543 (DSM 43111) D L S L P R L I A T E D G C S L G E A V R R T V D I H D E L M H A F E A E A A A L A - L T G S P E L R R F L W G L W A W L G G S R E W H A R
YP_347573 (Pf0-1) D F K L P Y L L A A R H N C S L Q E A I D L A A D I H D A V M E E Y E R L H A T L M K G T R S P V L R R Y L T G L S T W I G G N L E W H K H
YP_001105919 (NRRL 2338) P F N L P A V I A A E E R I A L D E A F V R A C E I H N E L I Q A L E E A T G H A A S A L A D P A L S R Y L T G L W S W L A G S R H W H F T
YP_003510780 (DSM 44728) D Y S L P G V L V A Q D G C T L Q E A V D R T A E L H D E L V R F V E S E S A L L S - A N H T P M L A R F L T G V W N W M G G G K E W H A T
CAJ89344 (ATCC 23877) H L N L P V V I A E R E R L S E R D A Y L K A V E V H N E L Q H A F E A A A A E L A K A C P L P T V L R F L K G V A A W V D G N H D W H R T
ADI12075 (BCW-1) H L N L P V V I A E R E G I S D R E G Y L K A V E V H N E L M H D F E A E A A A L A A A C P V P S V Q R F V R G V A V W V D G N H Y W H Q T
ZP_08234965 (XylebKG-1) H L N L P V V I A E R E G L S D Q D A Y L K S V E I H N E L M H A F E S E A A A L A A A C P V P S V Q R F L R G V A A W V D G N H H W H R S
NP_733742 (A3(2)) H L N L P V V I A E R E Q L C E R D A Y L K A V E V H N E L Q H S F E A A A A D L A E A C P L P P V L R F L R G V A A W V D G N H D W H R T
ZP_05801975 (ATCC 33331) H L N L P V V I A E R E G L S E R D A Y L K S I E V H N D L M H D F E R E A A A L A E A C P A P G V H R F L R G V A A W V D G N H H W H Q S
YP_001822781 (NBRC 13350) H L N L P V V I A E R E G L S D Q D A Y L K S V E I H N E L M H A F E S E A A A L A A A C P V P S V Q R F L R G V A A W V D G N H H W H R S
ZP_05520932 (ATCC 53653) H L N L P V V I A E R E G V S E Q E A Y L K A V E V H N D L M H D F E A A A A A L A A A C P V P T V Q R F L R G V A V W V D G N H Y W H Q T
BAI77523 (NRRL 3382) H L N L P V V I A E R E H L S D R D A Y L K A V E V H N E L M H A F E A A A A E L A A D C P V P A V L R F L R G V A A W V D G N H D W H R T
YP_003486275 (87.22) H L N L P V V I A E R E R L P V R D A Y L K A V E V H N E L Q H A F E A A S A E L A E A C P L P A V L R F L K G V A A W V D G N H D W H R T
ZP_07279119 (AA4) V C N M V L Q I A A D R N C P V S E A V E T T V A L H N R I V R E F E E G H R A L L - A I P S P E L Q R F L A G V R D W M G G G F E W H A T
ZP_07290591 (C) H L N L P V V I A E R E G V S D R E G Y L K A I E V H N E L M H D F E A E T A A L A A A C P V P S A L R F L R G V A V W V D G N H Y W H Q T
ZP_05002186 (Mg1) H L N L P V V V A E H E G G D V R D A Y L K A V E V H N D L M H A F E A E A A E L A A A C P V P S V L R F L R G V A A W V D G N H Y W H Q T
CCA60397 (ATCC 10712) H L N L P V V I A E R E G L S E R D A Y L K A V E V H N D L M R E F E A E A A A L A A A C P V P T V L R F L R G V A V W V D G N H Y W H Q T
ZP_07604855 (Tü 4113) H L N L P V V I A E R A G I S D R E A Y L K S V D V H N D L M H D F E A A A A A L A A A C P V P S V Q R F L R G V A A W V D G N H Y W H Q T
ZP_07301463 (DSM 40736) H L N L P L V V A A E E R C G L K A A Y L K A V E I H N Q I M D A F E E E S A A L S A G - - S P L V E R Y A R G L A S W V S G N H E W H A T
ZP_05521425/6 (TK 24) H L N L P V V I A E R E Q L C E R D A Y L K A V E V H N E L Q H S F E A A A A D L A E A C P L P P V L R F L R G V A A W V D G N H D W H R T

51
Figure 5. Alignment of 2-MIB synthases. Highly conserved amino acid residues are shown in red. Highly conserved motifs are shown in boxes. Missing parts of sequences are indicated by question marks.
501 511
YP_003765432 (U32) N P - R Y Q - - - - - - - - - -
YP_003115314 (DSM 44928) N S S R Y K - - - - - - - - - -
BAJ32779 (KM-6054) N T Y R Y S L P D F W - - - - -
BAK26793 (NRRL 8178) T S - R Y H G E A T T R S - - -
ZP_04609212 (ATCC 39149) T A - R Y H K Q Q G A D P A - -
YP_003680543 (DSM 43111) S P - R Y H G A G T D - - - - -
YP_347573 (Pf0-1) S A - R Y H I - - - - - - - - -
YP_001105919 (NRRL 2338) T A - R H R A - - - - - - - - -
YP_003510780 (DSM 44728) S R - R Y H K Q E S D N S D A A
CAJ89344 (ATCC 23877) N T Y R Y S L P D F W - - - - -
ADI12075 (BCW-1) N T Y R Y S L P D F W - - - - -
ZP_08234965 (XylebKG-1) N T Y R Y S L P D F W - - - - -
NP_733742 (A3(2)) N T Y R Y S L P D F W - - - - -
ZP_05801975 (ATCC 33331) N T Y R Y T L P D F W - - - - -
YP_001822781 (NBRC 13350) N T Y R Y S L P D F W - - - - -
ZP_05520932 (ATCC 53653) N T Y R Y N L P D F W - - - - -
BAI77523 (NRRL 3382) N T Y R Y S L P D F W - - - - -
YP_003486275 (87.22) N T Y R Y T L P D F W - - - - -
ZP_07279119 (AA4) N P - R Y R S - - - - - - - - -
ZP_07290591 (C) N T Y R Y S L P D F W - - - - -
ZP_05002186 (Mg1) N T Y R Y S L P D F W - - - - -
CCA60397 (ATCC 10712) N T Y R Y S L P D F W - - - - -
ZP_07604855 (Tü 4113) N T Y R Y S L P D F W - - - - -
ZP_07301463 (DSM 40736) N T H R Y H L P N Y W - - - - -
ZP_05521425/6 (TK 24) N T Y R Y S L P D F W - - - - -

52
Figure 6. Phylogenetic dendrogram based on the analysis of amino acid sequences from bacterial 2-methylisoborneol synthases. For phylogenetic analysis the
neighbor-joining method was used. The amino acid sequence of the pentalenene synthase from S. avermitilis (accession number NP_824174) was used as
outgroup. Bootstrap values are given next to the tree nodes. 2-Methylisoborneol production is indicated by black diamonds, and no 2-methylisoborneol production is
indicated by white diamonds. The proteins from strains included in this study are shown in bold.

53
Figure 7. Genes for the biosynthesis of 2-methylisoborneol (2). The colour code is black: regulatory nucleotide binding protein, purple: 2-methylisoborneol synthase,
grey: GPP-C-methyltransferase, white: genes that are not related to the biosynthesis of 2.

54
A) Nostoc punctiforme SAG 60.79 (GQ287652)
Chitinophaga pinensis DSM 2588 (NC_013132)
B) 50
34
Streptomyces scabiei 87.22 (YP_003486275)
Streptomyces coelicolor A3(2) (NP_733742)
Ktedonobacter racemifer DSM 44963 (NZ_ADVG01000001) 99 Streptomyces ambofaciens ATCC 23877 (CAJ89344)
91
99 Herpetosiphon aurantiacus DSM 785 (CP000875) 48
„Streptomyces lasaliensis“ NRRL 3382 (BAI77523)
Roseiflexus castenholzii DSM 13941 (CP000804) 20
Streptomyces sp. Mg1 (ZP_05002186)
100 Haliangium ochraceum DSM 14365 (NC_013440) 26 Kitasatospora setae KM-6054 (NBRC 14216) (BAJ32779)
96 Plesiocystis pacifica SIR-1 (NR_024795) Streptomyces flavogriseus ATCC 33331 (ZP_05801975)
96 Sorangium cellulosum So ce 56 (NC_010162) 91 Streptomyces griseus subsp. griseus NBRC 13350 (YP_001822781)
20
100 Myxococcus xanthus ATCC 25232 (DQ768116)
Stigmatella aurantiaca ATCC 25190 (DQ768127) 100 Streptomyces cf. griseus XylebKG-1 (ZP_08234965)

34 Streptomyces hygroscopicus ATCC 53653 (ZP_05520932)

74
Rubrobacter xylanophilus DSM 9941 (NC_008148) Streptomyces violaceusniger Tü 4113 (ZP_07604855)
47
100 „Streptomyces bingchenggensis“ BCW-1 (ADI12075)
Catenulispora acidiphila DSM 44928 (NC_013131)
Actinosynnema mirum DSM 43827 (NC_013093) Streptomyces venezuelae ATCC 10712 (CCA60397)
82 99 Streptomyces sp. C (ZP_07290591)
92 Saccharopolyspora erythraea NRRL 2338 (NC_009142)
99 64
Amycolatopsis mediterranei U32 (NC_014318) Streptomyces viridochromogenes DSM 40736 (ZP_07301463)
Saccharopolyspora erythraea NRRL 2338 (YP_001105919)
Kribbella flavida DSM 17836 (NC_013729)
93 Pseudomonas fluorescens Pf0-1 (YP_347573)
100 Micromonospora olivasterospora DSM 43868 (X92613)
Micromonospora aurantiaca ATCC 27029 (NC_014391) Streptomyces sp. AA4 (ZP_07279119)
76 100
Micromonospora sp. L5 (NC_014815) Amycolatopsis mediterranei U32 (YP_003765432)
100
100 Frankia sp. Eul1c (NC_014666) Catenulispora acidiphila DSM 44928 (YP_003115314)
Frankia sp. EAN1pec (NC_009921) 42
89 Stackebrandtia nassauensis DSM 44728 (YP_003510780)
Frankia sp. Ccl3 (NC_007777)
93 Nocardiopsis dassonvillei subsp. dassonvillei DSM 43111 (YP_003680543)
100 Frankia alni ACN14a (NC_008278) 85 Micromonospora olivasterospora NRRL 8178 (BAK26793)
100 100
60 Stackebrandtia nassauensis DSM 44728 (NC_013947) Micromonospora sp. ATCC 39149 (ZP_04609212)
Thermomonospora kurvata DSM 43183 (NC_013510)
68 Nocardiopsis dassonvillei subsp. dassonvillei DSM 43111 (X97886) Outgroup
Streptosporangium roseum DSM 43021 (NC_013595)
76 Planobispora rosea JCM 3166 (AB028654)
0.2
Streptomyces cattleya NRRL 8057 (FQ859185)
Streptomyces albus subsp. albus DSM 40313 (AJ621602)
„Streptomyces bingchenggensis“ BCW-1 (CP002047)
61 Streptomyces violaceusniger Tü 4113 (NZ_AEDI01000202.1)

100 Streptomyces hygroscopicus subsp. hygroscopicus NBRC 13472 (AB184428)

Streptomyces ghanaensis ATCC 14672 (NZ_DS999641)
Streptomyces griseoaurantiacus NBRC 15440 (AB184676)
Streptomyces pseudogriseolus NRRL B-3288T (DQ442541)
Streptomyces griseoflavus LMG 19344 (AJ781322)
Streptomyces ambofaciens NBRC 12836 (AB184182)
Streptomyces viridochromogenes NRRL B-1511T (DQ442555)
Streptomyces sp. SA3_actG (NZ_ADXA01000222)
Streptomyces coelicolor A3(2) (NC_003888)
92
Streptomyces odorifer DSM 40347 (Z76682)
77
Streptomyces arenae ISP 5293 (AJ399485)

Kitasatospora setae KM-6054 (U93332)

Streptomyces scabiei 87.22 (NC_013929)
Streptomyces clavuligerus ATCC 27064 (NZ_CM000913)
100Streptomyces griseus subsp. griseus NBRC 13350 (NC_010572)
Streptomyces cf. griseus XylebKG-1 (NZ_ADFC00000000.2)
Streptomyces peucetius NBRC 100596 (AB249907)
Streptomyces sp. ACTE (HM235473)
Streptomyces flavogriseus CBS 101.34T (AJ494864)
Streptomyces filamentosus NBRC 12767 (AB184130)
98
Streptomyces pristinaespiralis ATCC 25486 (ABJI00000000)
93 Streptomyces venezuelae ATCC 10712 (FR845719)
Streptomyces avermitilis MA-4680 (NC_003155)
Streptomyces sviceus ATCC 29083 (AB184559) Figure 8. Comparison of a 16S rRNA gene-based phylogenetic tree (A) with the phylogenetic
tree based on the amino acid sequences of geosmin synthases (B). Groups of related strains
0.1
Streptomyces lasaliensis JCM 3373 (HQ537060)
Streptomyces griseocarneus JCM 5010 (AY999766) and their geosmin synthases are shown in the same colour. 55
Figure 9. Mass spectrum of two unidentified compounds (A and C) and similar mass spectra of the known compounds zizaene (B) and prezizaene (D). Due to the
similarities between the mass spectra and due to biosynthetic considerations the unknown compounds are suggested to be epi-zizaene (A) and epi-prezizaene (B).
These compounds have the same stereochemistry as the main product epi-isozizaene of the respective sesquiterpene cyclase.

56
Figure 10. Partial total ion chromatograms of headspace extracts from albaflavenone producing actinomycetes. Continued on next page.

57
Figure 10 (continued). Partial total ion chromatograms of headspace extracts from albaflavenone producing actinomycetes (A) and mass spectra of the tentatively
identified sesquiterpene alcohol 72 and the known compound zizanol (B).

58
Figure 11. Structures of further identified terpenes from bacteria that are produced by unidentified bacterial sesquiterpene synthase homologs.

59
References

[1] R. P. Adams, Identification of Essential Oil Components by Gas Chromatography/ Mass Spectrometry, Allured, Carol Stream,
2009.
[2] D. Joulain, W. A. König, The Atlas of Spectral Data of Sesquiterpene Hydrocarbons, E. B.-Verlag, Hamburg, 1998.
[3] Q. Wang, Y. Yang, X. Zhao, B. Zhu, P. Nan, J. Zhao, L. Wang, F. Chen, Z. Liu, Y. Zhong, Food Chem. 2006, 98, 52-58.
[4] C. A. Citron, R. Riclea, N. L. Brock, J. S. Dickschat, RSC Advances 2011, 1, 290-297.
[5] J. A. Pino, J. Mesa, Y. Munoz, M. P. Marti, R. Marbot, J. Agric. Food Chem. 2005, 53, 2213-2223.
[6] J. S. Dickschat, N. L. Brock, C. A. Citron, B. Tudzynski, ChemBioChem 2011, 12, 2088-2095.
[7] S. Hamm, J. Bleton, J. Connan, A. Tchapla, Phytochemistry 2005, 66, 1499-1514.

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