Korean Journal of Life Science Vol. 12. No.5. 632~648, 2002 a4 Polyketide O|AHCHAHES9| seta 20! ‘gata ahat Bah geen etal ee} Baa Se Ht, AeA a Biosynthesis of Polyketide Secondary Metabolites Yoon, Yeo Joon and Sohng, Jae Kyung! University of Ulsan, School of Ohemical Engineering and Bioengineering San 29, Muger 2-dong, Nam-gu, Ulsan, 680-749, Korea Surmoon University, Division of Natural Sciences #100, Kalsan-ri, Tangjeong-myeon, Asan-si, Chungnam, 335-708, Korea Abstract The term polyketide defines a class of natural products synthesized through the successive condensation of small carboxylic acids, which results in products containing multiple carbonyl or hydroxyl groups, each separated by one carbon atom, as in the structural element CH:C(-O)CH:CH(OH)CH.C(-O}. Plant flavonoids, fungal aflatoxins, as well as hundreds of compounds of different structures that can inhibit the growth of bacteria, viruses, fungi, parasites or human tumor cells are included in this se group. Some of antifungal polyketides also have immunosuppresive activity Polyketides can vary widely in structure, and the diversity of polyketide structures reflects the wide variety of their biological properties. This review focuses on the biosynthesis of polyketides and recent progress in combinatorial biosynthesis of new hybrid polyketide compounds. Koy words ~ polyketides, Streptomyces, combinatorial biosynthesis Af Polyketidex) S}¥0]2 multiple ketone 23, (CH-CO), # Aa NE BAM} |]. CEM FA acetyl, pro- pionyl, = butyhCoas YET sto} Se wse AA GA a AS Abela FH ketone (Pketone) HEE ke- “To whom all correspondence should be addressed Tel: 04530-2246, Fax: 04151-7425 E-mail: sohng@omega sunmoom.ac kt 2 /-Bs}s4812) tones} Sl¥o] Gzioe Baste eolae ae sit B10}, polyketoneol Slt HES] keto 15-2 Mal FS) SPS AA Ge] 714) HE 715718. ACE (Scheme 1), Polyketide AIG AS 2el= HE 4741 1074 bui- sok On On \ ai ee Scheme 1Polyketide LMHS 8A ling blocks) 4 Wao} PAAAR, Ae Mole wet Japa abe. Polyketidesd HE ol7t AY Bed 8 ALE GE FH) polykelider| aE] Aaa @aaol 2 BOVIS oF ST AE ss] BH BHU AR sich. YHA erythromycin, tetracycline, methymycin, 444] doxorubicin, neocazinostatins, 82% ‘all rapamycin, F506, antiparasitic ala] avermectin, emadectin, 217A) amphotericin, griseofulvin, A13E B19 lovastatin, compactin 12]32 $82 monensin, tylosina] =2 292} ao]o} rapamycins} FKSIGE: 13.5) signal transduction #241 AMESI71S she Fig. 1) [19]. APE polyketide: Yat Fo] Base Fae a ‘GEZA| Coenzyme A-activated carboxyl acide] S4}2. 2 Aa Bey GY FRE ole polyketide) st BoE OG ZF poly-bketone SUAS AA HY Je SEAS AR Gl ABA, oleh Boles AAS 9) SAS oS. HAbSHh Polyketides} faty acids) $34} © CoAB HE] ao] a acyl APE Garter unije] f+ etoacyl synthase (KS)9] 218} Wo] AabeIch. AE BE 9} 2249} Gee carboxylated acyl ALG (extender unit) ©] CoARI} acyl cartier protein (ACPI. BAA, Ate A ATM} Le F MS acyl TES decarbosylation =} 24 SMA, Fatty acd AYA BF Sree Ah] B- carbonyl® methylene®:2 gato} o]s]2)eh, poly- otide $829] BF S7HE AES] 8 carbonyl 10} 2 keto THOS YAU, BE Fol lal, hydroxyl, oF AGT HL methylene 715712 Aolsleh. Polyketides] 2X} €2] Bol, MA AE, stereochemistry 5! cyclization 2 PKSo aie} cfs, cyclized polyketdesd} #4212 2 G2, methylation, acylation, glycosylation 3 abe} ol she} Zar; ASA VAY Sey Polyketides} fatty aids] S82] FRE SA eat ee Salad dee ual aA Sb] SHS UE + Set (ig 2. 8 At DS of] 48H actinorhodin, granaticin ¥ tetracenomycins} @ & bocterial aromatic polyketide YebJsh= PKSS} Dacterial 3} plant FASE Li) 22 active site 95) 4} gkch. 72}9} adive site= Sa RAE FSI} g oF Wad TEE SE fungal] FAS 3 Gmethyts alicyclic acid synthase (MSAS)$} 2:¢ fungal PKS=4} 84} SE FAY polypeptides PYsIo} YH, SYA active sitet domain ehz 9sIo Qe}. 4H (autienzyme) he Fr dS Veh sled ad eq dal Be WS FMC AS 45) active ster} sIt He] SI 172 2 8 AS) active site GH WHALES 788 BSE 47} SO) F Wat THO) 7d Seb. A Wa ABE F WAS de] TE 4p HAA|D modular PKS 9} Sol ale. CaNSS Waa wase Av, 22}9} active site 2}o] HAWS ASS, active sitet module JessI9} $20}, 7 module B24} S91 FGA Melo POs active sites AHIR Sle aL 2A LHL chalcones} stilbene #4] HA€) PKSE GH 72] eae FAS Sz, Hot Bae PRS FASS #4} Aol de Wo. 1S Ge Bedee de qyase Bojaleh. o}]5} G2 polyketide synthases} nonrib- ‘somal polypeptide synthase) hydrid@ Wel @al2}} reduction WE eh Go] SEAS (7, 7) 2) Aromatic PKS9 heterogeneous Zt Aromatic polyketide AYES WHY alah 23S F eA ses G4 ede Hee Rae ss Slant He LU Sol Vola Bspo|ch. Aromatic polyketide 49 SUAS WAS AS HL polyd -ketoneo] 445171 GE Sus YY Bro] 71a ae a4 Selah Sa Bee] Teele aaa in vitro 2¥o] B7hsaech. olst Be Byes] Gale MZG PKS FAAS PAA AND TAN ASE BES) FAA AG HAs Seloleke Tyo A F4HIAD Ho4oe W2e PLS polyketides BAT Me PAS AA Ach. AA PKS HAAS oleate] Mola Ase Sesto] vad deat go] y BR MO} (Table 2, Fig 5) @ 71% S4q poly-f-ketones| %o]= minimal PKS 2}8 USC. Minimal PKSe] FA) VAIS SHR ACE Ie B44 doll AE FA] a we ‘ad $ 20} (Table 2, entry 1~6), KSa3} KSBE SL BSG BSA Me Bet FEMA (Table 2 entry 7-19), DKRE 2el7} HE poly--tetones: Veh ea 22 94 PLS] carbony7]E JESS 71Z Be) ‘att (Table 2, entry 20-24), ® Poly--ketones] GUE KRo] Me BF, UaAos EAE cyclase] FHA AeA] BBS} (Table 2, entry 25-28) AA) 1 G29} 9 Ge Te] 2 32) ofsiaiog 7a Use| dade aaa oh KRO] Re BP BTS Masi CLA aa Ay eiee @ MZY PKS] BH 2a} EE 3A} Dese RAS ol oh #24) BLAM ely age BEALE Typell PRS} 915}2] 4448 aromatic poly- ketides) PAE AYA Boleks TARAS 88 A EA aN} AAR o}oHVe: Typell PKS Halzho} ap 714% $42 oS ZA (combinatorial biosynthetis) oz Mati] Ae P29 EE Heyl ae So WAL ASE eld. ASE 729) aromatic polyletide? Qa] ME AE ol Ve QA Be of BORE PKS 74 HAS UY He] qylascs 1S Vol. 12. No.5 (2002. 10) / 637BE SF Mint oance nee mm BOO Oo, Opens dita ooh. S88, HS - | Actinorhoain Fig, 4. Biolsynthesis of actinorhodin. Table 2. Polyketides produced by recombinant PKSs. Entry Koo KS} ACP KR Metabolites Reference ‘ACP changed at act att SEK 31 2 at ack SEK4 9 3 act act act acta VL DMAC B 4 at act grt act at VILIV DMAC B 5 act acl fren act act VILIV DMAC 2 6 at act tomcat VIL DMAC B KSotp change? fon tom act act at VILIV M20 B 8 fn fen act act, aVILIV. DMAGRMI8,RMI80 2 9 act act tem tom uw » 10 ops des tom tem TemF2 B 0 jad jad tom tem)N TemF2 4 Ka change,12 gm act, act, achat VILIV DMAC B B ton act, act acta VILIV DMAC B “ fren act act act, act DMAC » 6 at tam tem temnjN - 39 KSB change.16 ft gmt act acta ILIV DMAC 2B wv fom act ho tomjN Tem, UWMI(trace) 2” 8 ect tem ack acta VIL - B 19 at fren act achat ILIV : a KR change.20 ton tom stomach RMI 31 a fom fom tom act tern N R206 0 2 fem tomtom dps tem RM20b,TemF2 3 B fem tomtom fad tem M206 4 m ofc oe eat oie M26 2 Folding change25 tem = km tom tem]N TemF2 9 2% at att tamjN RM? 30 2 fren fen fron temJN Pe B 8 fom tem tom act VILIV SEKISSEKISb 3 68 / BAHIA)Polyketide o|a}4 229) “84h Be “Os. Fig, 5. Hybrid aromatic polyketides produced recombinant PKS, FEUGI Ge GI Fold lz Ad. Hig. 6 oA Be ¥}S} to] AE CHE 24} GAHE ao] obzyet a eae Sots RAE aA F SHS Golan ole $414} (4SE9} tetracenomycin2} tom9} actinorhodin9} aciVi-orfl) 22} 38 BBA eH hybrid antibiotics AG + Ne esol ST sac. ols} BE wyol da Aisle sHMeS Fae a Fad aA de 7) G7] Met FH] VES WYAE oxidase, sugar transferase, methyltransferase $2 Rap SMD F SLE SRE SE Bole. 3) Modular polyketide synthase Mactoide polyketidet= ©}7)'5 #29) type 1 PKS 5] A Set 4 Fol BA, GAldehydration FE eldeh- ydration-@41 39 Catch} 7A] macrolide ek 3} GE (highly reduced macrolide)Z4 4194. Type | PKS ©] 919] 2S polyketidet= aromatic polyketides#-} poly- Preto Szat Bch also} See Boh oe} ANS A (keto, hydroxyl, enoly $2 alkane}$: 7H S}KZ= East] macrolide, macrolactam, polyether, pol- yene $ He} be] BEB] ols} BE aS AM Ba dt} macrolide polyketidee) erythromycin aglycone (eryA}st avermectin aglycone (er) BBB ABER aah aaa) wala leh ARYA Saute) P43} ee aa aald hee, 4 2% moduleZ#=} @ units} polyketider} 9514 fatty acid 42} HHA STO}, 222} modules HS} S ARPYE 718 @elet polyketider? PAA. Module fA gees ade] gle) AEH. eel complex polyketides} 499 she, 44shiah Suet acyl group (acetate, propionate, butylate, m-C;N unit)®} AG, PEAS WY FA age At So} aol, & cycles} sho] BA Balada gL polyketdet ACPS 712) #, 8 a¥4) Hola} modules} KS3} Vol. 12. No.5 2002. 10) / 639ee ea “Tetrwcenomycia nes Michramyein = runorabica oe = og Figure 6, Structures of aromatic polyketides involved in cyclization step. acyl group unit (extender)s} West] 2 G8 OA ae o] AGT. ©] HS WHOZ TYTN, 2 elongation Gali ABE modules] aelsie} laa Cares} 7|ol SER wip) ALC, Sectaropolyspora erin 44) 4AS}E erythromycin] 32121 of 24) aglycone 40 / 482485 8)2) AWA FAA eA HAA) AAA] BT SISIG. 1S Galt WASH 6AEB (6-deoryerythronolide By] PAs} aheFsHs 37H] eryA ORF (eryAl, eryll9} erlll) #22H2 modulesle}T Be] WASTE synthase univ OE FY} RDF aris F Wks FASPolyketide 14}: 839) $3 ©} a, $2) 6deoxy-erythronolide BE YBAEAT Bal SHE exy PKS 4 AFH2 32 kb of A} Oh sal S QA AAT YT, 6deoryerythronolide B (6-dEB) ] erythromycin Avia] 81:2 Galol eeiahe non-PKS Fale Bel AIT LOM, al erythromycin B GABE HAAS oO kde HLA [20] Fig. 7. 2 Module 4 stoheh S79} a7} volo}, 3) BALE WA BLE olFolal Habel macrolided & GED AAA Set (Fig. 8. yA) Sl LE modules} = SPP else ketosynthase (KS), ATS} ACPA} ABIES domaine] FPHo} 3x, Pearbons Wa) = 715% 7#al domain [KR, dehydratase (DH), enoylre- ductase (ER): 2} modules} wz} Be} LAS}o} she. ap module® #23}. DEBSI 9 JE} starter unit@l prop- fonateol sfSit2 AT®} ACP domain®.2 798 loading module®} 3151, DEBS3 % 2240} chain extensions} & BE Yeu TE domaino@ Ale end modulee} 2 5. eR moduledol= KR domaine} ge} nonfunc- tional. #7}5)"] DEBS32} 2 module? thicesterase % 191 S}94 polyketide chains} lactonization® &214]713, 2 AS} RASH macrolides release 7} 7153-2 7+ AZ SIC. modules of} methylene groups YAH UA E sete] BA domaing @apal 7D sk. ‘t}2}4] module 42} erythromycin PKS2] tH2 module AF ©}} 3}0]430] ATS} KR domain AFo]} th= DH, ER 2H KR) SeohFeh cep 715O] Welle gle a $24 Gh. 19] AAA He Halas] $9 domain © FAS (gene disruption)é}e} 2} domains] 7} a qaalz, BY hie Hat 759] Felabs olgsta} FE WHE F 1SE BITE Ue ole de FEROS Gag FAA Jal Alsjojop spe dal Z Bol MAIC GLE] PRSE 7Idl de Solo} go} Al7| AEA o| BUS BA polyketides| FAVBE He) olBsin wet (Fig. 9. 4) Module polyketide #2121 Reprograming Exythromycins} PKS #ralaHe chain extensiono} 2} moduleol 2]a) GSI IS BF sz, Subse A 29) proteing ‘cassette’ Belslal 7 alc. Ee olel FAOR UFO] | af cassettes] EYES ZAYOLH gt cassette ¥E} CHE cassette®.9] growing chain2] °}'$0] Wee AE VF a. @ Module #:2}2] 282 St polyketide FAW Module 745) 22% Eel] 9412 polyketides) 7 EE EACOSM complex polyketide} module types} Siskel I Saez ahaa werisot slat GATS WSR + sd. DEE Aeee delay 7s RAF, modulel3} module2S ¥ tet DEBSI2} DEBSS2} thiesterase (TE}S ARS} tiketide 4 4 Ae UEFA I AAA) olFo}] aE a ©} SBS|AAF [16], Moduledo} TES hybrid 2121 RAs} DEBSIG. HSH HAAS S. eolclor CHI) Baal BAA AF tetroketider} A519} [27] (Fig. 10), module 59} module 6% hybrid 2)2] AAS} DEBS! 2213. DEBS2 Haz}s] PaVeA Ze] GABBA ring 27]7} SE SHES] Adee AS BolgoeA 7, 18] Te} at ie rao] epee za} module A BAS Fal macrolide polyketides] a7 BV + Se AS SUA Fig 10). Wee 4 DEBSIATE hybrid $28 18% 34 in vittools) avER propionyl CoAd}’l acetyl CoA, butylyl CoA EE ZLmethylS-hydroxybutyl Nacetyleysteamine thioester7} PAL ASI, ZEA NADPHE a7hsh] Be W_ Gethyl-2-hydroxy-3,5-dimethyl-2H-pyran-2-oneo} 842 ATE AS SUVSI4 FEE FH2I erythromycin SAS UE $1 ASAE Bol FH. ® Domain Ea 218 polyketide F209 Exythromeyin PKS 212} modulel, 2, 68} AT dom- ain rapamycin PKS % malonylCoAS S932 AT domain&.2 21G33 sf malonylCoA7} extender units. SIE erythromycin VFL aisle Fig. 1). Table 3 4A] BE vist gol Rar] ai) SHS extender unit 144 4 91 AT domaine] #2151347] "AE polyketide lactone ring?) AU Ea) 2H) 2.2 OAH) FEE SF NS Rol. Avermectins| 4 starter units isobutyryCoAS} 2methyl. butyryl CoA BY O42 HOS FHA HES VGH a Vol. 12 No.5 (2002, 10) / 681844 - Ene eryPKS genes, onal coal > enyaltt o 10 20 30 0 50 60 kb AE non PKS genes ton PKS genes I~ ve exBieyc epay eB ency mE ct” ena rg tor ent "entvi ent” at ORF Fig. 7. Genomic map of , erylhraea involved in erythromycin biosynthesis. a wal (sar) rast DEBS i fl Polen i ra sists) betel ie Gebetst J islet babslal betel z 1 T T T 4 { I \ =o no 0 mo 5 o 0 on 1 S6 on )~on on " on Interment in Chain ed oH oH 0 Extemion Cycler \ on H \ é ” Deoxyerythronlide B 9 on on Fig, 8. Biosynthesis of 6-deoxyerythronolide B 642 / gaits]Polyketide olAbciAHSa) 4h ell ) coal, ) Ss -DEBs— Fig. 10, Intermediates produced by hybrid PKSe in which DEBS TE domain was replaced. Vol. 12. No.5 2002. 10) / 64384S Fae aaa oH oc ( . Das} ‘Pou PKS modieation ° seucenes tH SiS ‘KOSOIS.20h RIH, ROH Fig, 11. New compounds produced by hybrid PKSs in which AT domain of module 1, 2, 6 of DEBS was substituted by malonyl-CoA specific rapamycin AT domain, Table 3. Polyketides biosynthesized by Type I PKS. ‘Compound Starter Extender ~Moules ORF Reference Erythromycin PropionylCoA __MethymalonylCoA 6 3 0 Tylosin PropionyiCoA ——_4MethyimalonylCoA 7 5 2 2MalonylCoA ‘BthylmalonylCoA \Niddamycin —-MalonylCoA——‘IMethyimalonyiCoA 7 5 Senelestis 5 4MalonyICoA ‘1BthylmatonylCoA ‘HydroxymalonylCoA ‘AvermectinIsobutyrylCoA or SMethylmatonylCoA 2 2 Savers B 2-Methyl ‘7TMalonylCoA butyrylCoa, Rifamycin AHBA 8MethylmatonylCos 10 5 Amediterrne 1 2MalonylCoA Rapamycin ‘Shikimate ‘7MethyimalonyiCoA 6 3 Shygroscopicus 8 7MealonylCoA, __Pipeoolic acd i & ©} erythromycin module 19] starter unit 9] BE vhs} ge] erythromycin} starter unit7} ost HE AT domaing avermecting) starter units SI3H= AS. isobutyryICoAS} 2-methyl butyryICoAS oa F AT domaine fal} Al#o| Asse. 2B 7 Ze Wastsieh 644 / t2}chs3)Polyketide 13-229) 8G SAGA VE domaing SUE BHIE I AT FE HH ABET OE domain wee IaH Be UF ole Far 28 GIF USS Ue Se A oleh. ol Se} exythromycin modiule23} KR domaine] rapamycin PKS false] 2af8}= DH/KR sE& DH/ ER/KR domaine 2 ABSIRE BF VAI SHAS] HANA (Fig. 12). 5) Combinatorial biosynthesis SUS4d Yas 4U9 ded G8 aol 20g Ale AG Sate WSIS VTS Whsto} Ado] EAs] BE VAIEAS Buster safe] 2 Aad dea Peeve. 2a MS gaa Wyle T2agel Nes PISAEAS ANS fra ae B48] 724 UGHS Avaeie wos al 7b $2 polyketide AW] PIEA FREI Eas Zt. SH M2 Val Se FSIS Ael poly- ketide P4Ea UY Hae Ase Aas sal al BAA A7} PKSo Saha} GIGS SEEM erythro mycino] Qc. 713% 7H) PKSE S0i}8S HES aon a aes Fig. 12. Biosynthesis of tetraketide produced by replacement of DEBS KR2 domain with DH/KR domains and structures of hybrid macrolides produced by feeding acylNAC to DEBS KSI null mutant BEY} RE VALS EAs] ALA module PKSe}at = BUC. module PRSE: SE AWAEIRA (FAS) © pla AG HAUS BolD] ARHIEA (Ketos- ynthase, KS), o}42 eat: (acyltransferase, AT), 2478] 2 (dehydratase, DH), 18-8182: (enoylreductase, ER), AEBURA (ketoreductase, KR), o}2aS Ma (acyl carrier protein, ACP), E.@AAH23:R2 (thioesterase, Te) & ES FAS Sage PVE LF VHT Up SATE BE FAS OS BESS WE AbG sth NS} Bt GAS FI }A-COA (acetylCoa)s} wz. ¥W-CoA (malonylCoA)2] + HSS Soh Sou) esl E PKS F AS] Be, A AS] HA BL Ao] Wa Sal 2 Fad A2BAs Azo] SY USS F Ae] BE A Soh Oth we}y pezbol= SHS] PARA BA) SH OS] AMAAEATS Hele As Ha At BAS] VRE FAs} Sle AS ASU + ale SE FASS, Va PKS] SE HE AOlBS BE PKS Hike BA YP OMYolo. SH TASE Ese 2 S48 (sae Le Pkse Ba BIH F24 We AS, MSA), Ba OS GE TL SE = dae F ae. 2 Ba PWS oly) at (KOSAN biosciences)o} 4 KEV oe] Helsigia, AB BAe sede = Fo] SG HASHES ESSER seh [Ml]. o] YY 22 294 77} Sa = 2 Ae ae] laa a Aa) Bald Fee AGA AF AsslD wed, o] = polyketided BAe 2H V4 AY Roads 34 Bad Agen Dold Wey GAVE ated de 0] HS 7H A BHM polyketidets HY a7 2] QE} Beye AS 37194 polyketide7} W2b@ Zest, Bad ME de 752s BAN seh P27) CE polyketider} Ba oleh a] st Basle W40] S7L polyketides BIE F sich AB Se} ryt hromycins] 34 2% c}243 (combinatorial potential) PKSo} le] BA S FRE AGRA ATS] R29 #8A ARM BE FR Moe 440] Phestch F CP=ATL x (ATe x 4)" Ie] ZY E74) (loading domain) 9) Soe AT FR ATI GANG Dole VIE SYACLA(malonyliransferase) FA ATy Becarbon Vol. 12. No.5 2002. 10) / 645BS oA 8 PR 4, eZ Mo) WEE ASEM. Exythromycina) BH Mr6, AT=3 2. Th FobE ele} CP=10,000,000 o} AACE. polyhetide= VEO] OBS] MEd Ze + de BARS acd. ee he UES] BAAS Ve Tala ale $9] GALE ASAT deorysugar)z 9} Fela] 2H, Bal Bay ME SAGE 2000) 72 o} 240] deal lc. wep) olSat AE BO GSE a AES GAG + Vd. AB felt ALE Sireplomyers venczuclac?} Bak BPA methymycinel SAG A Jol HE AS OTD Meh 1H6 6) azead 1. August, P. R, L. Tang, Y. J. Yoon, S. Ning, R, Mueller, T. W. Yu, M, Taylor, D. Hoffmann, C. G. Kim, X. Zrang, C. R. Hutchinson and HG. Floss. 1998. Biosynthesis of the ansamycin antibiotic rifamycin: deductions from the molecular analysis of the rif biosynthetic gene cluster of Amycolatopsis meiterraei 699, Chom, Biol. 5, 69-79. 2 Beck, J, S. Ripka, A. Siegner, B. Schiltz and E, Schweizer. 1990. The multifunctional 6-methylsaic acid synthase gene of Pencil patulu. Its gene stt- uucture relative to that of other polyketide synthases. Eur. |. Biochem. 192, 487-498, 3. Bedford, D. J, E. Schweizer, D. A. Hopwood and C. 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