TP CH SINH HC 2013, 35(4): 469-476

TI U HA THNH PHN MI TRNG TO KH HYDRO SINH HC

CA CHNG VI KHUN K KH Thermoanaerobacterium aciditolerans Trau Dat
PHN LP TI VIT NAM BNG PHNG PHP P NG B MT (RSM)
Nguyn Th Yn1, Li Thy Hin1, Nguyn Th Thu Huyn1,2*
1

Vin Cng ngh sinh hc, Vin Hn lm KH & CN Vit Nam, *huyen308@gmail.com
2
Trng i hc Nguyn Tt Thnh, thnh ph H Ch Minh

TM TT: Th k 21 l k nguyn ca ngun nng lng ti to, trong nng lng hydro sinh hc l
la chn sng gi c th thay th ngun nhin liu ha thch ang ngy cng cn kit. Hydro sinh hc ang
c nhiu nc quan tm, bi n c nhiu u im v c to ra bng con ng ln men vi sinh vt.
Chng vi khun Trau DAt phn lp Vit Nam c kh nng to kh H2 sinh hc trong iu kin ln men k
kh. Th tch kh cng nh thnh phn kh to ra ca chng vi khun ny ph thuc nhiu vo cc yu t dinh
dng v iu kin mi trng nui cy. Khi nghin cu nh hng ca tng yu t dinh dng trong mi
trng nui cy cho thy glucose, cao nm men, FeSO4.7H2O c tc ng mnh n qu trnh to kh H2 ca
chng vi khun Trau DAt. S dng phng php qui hoch ha thc nghim bng p ng b mt da trn
kiu tm phc hp vi 3 bin l glucose, cao nm men v FeSO4.7H2O, xc nh c s tng tc gia
cc yu t nu trn cng tc ng n th tch kh hydro sinh hc do chng Trau DAt to ra. Phng php
qui hoch thc nghim cng xc nh c hm lng ti u ca 3 yu t ny cho qu trnh to kh hydro l
glucose 12 g/l; cao men 2,5 g/l; FeSO4.7H2O 400 mg/l. Th tch kh thu c khi ln men chng Trau DAt
vi cc thng s ti u ni trn l 176,02 ml/150ml mi trng. Ln men theo m vi th tch 600 ml mi
trng trong iu kin ti u thu c 420 ml kh hydro.
T kha: Hydrogen sinh hc, RSM, vi khun, Vit Nam.
M U

Hydro sinh hc c coi nh ngun nng

lng thay th trong tng lai do khi t chy
sinh ra nng lng ln, khng to ra kh CO2, v
vy, khng gy nhim mi trng. Nghin
cu sn xut hydro sinh hc c nhiu quc
gia trn th gii quan tm, trong c qu trnh
sn xut kh hydro sinh hc t cc vi khun k
kh [1, 6, 10]. Cc kt qu nghin cu qu trnh
ln men thu kh hydro t vi khun k kh cho
thy qu trnh ny b nh hng bi nhiu yu
t dinh dng cng nh yu t mi trng. Do
, ti u ha cc thnh phn mi trng nui
cy nhm nng cao kh nng to kh hydro ca
cc chng vi khun rt quan trng [1, 2, 3, 6,
11]. Gn y, nghin cu ti u cc yu t mi
trng ln men to kh hydro s dng
phng php qui hoch thc nghim bng p
ng b mt c nghin cu nhiu [7, 8, 12].
S dng phng php ton hc qui hoch thc
nghim cho php xc nh c iu kin ti u
ca a yu t mt cch chnh xc v mc
nh hng ca cc yu t n qu trnh to kh
hydro trong cng mt thi im. Thm vo ,

phng php ny cn cho thy vai tr tc ng

qua li gia cc yu t trong cng mt thi gian
nui cy vi s ln th nghim t, nh gi c
sai s trong mi ln thc nghim theo cc tiu
chun thng k, xem xt nh hng ca cc yu
t vi mc tin cy cn thit, tit kim thi
gian v chi ph m vn em li hiu qu mong
mun. Bng vic s dng phng php qui
hoch thc nghim, nhiu tc gi ch ra s
tng tc qua li gia cc yu t mi trng c
tc ng n qu trnh to kh hydro. ng thi,
s dng phng php ny cng xc nh
c nng ti u cho cc yu t dinh dng
v mi trng cho qu trnh ln men to kh
hydro ca cc chng vi khun [7, 8, 9, 12].
Trong cng trnh ny, chng ti trnh by kt
qu ti u ha ba yu t glucose, cao nm men,
FeSO4.7H2O trong mi trng ln men to kh
hydro sinh hc t chng vi khun k kh
Thermoanaerobacterium aciditolerans Trau
DAt bng phng php qui hoch ha thc
nghim p ng b mt (Response Surface
Methodology-RSM) v kt qu phn tch thnh
phn kh khi ln men th tch 600 ml dch
nui cy trong iu kin ti u tm c.
469

Nguyen Thi Yen, Lai Thuy Hien, Nguyen Thi Thu Huyen
VT LIU V PHNG PHP NGHIN CU

Chng ging v mi trng nui cy

Chng
vi
khun
k
kh
Thermoanaerobacterium aciditolerans Trau
DAt c phn lp trn mi trng chn lc
NMV t mu phn tru ng Anh, H Ni [4].
Mi trng nui cy chng vi khun Trau
DAt l mi trng NMV [4].
Phng php
Nui cy vi khun k kh trn mi trng
chn lc NMV.
S dng phng php qui hoch ha thc
nghim bng p ng b mt thit k ma trn
thc nghim [4].
Xc nh th tch kh hydro bng phng
php th ch nc (water displacement method)
[4].
Phn tch cht lng v hm lng kh
hydro bng my sc k kh GC-TCD (Thermo
Trace GC-Thermo Electro-USA) vi phng
php th EDC VI-003 GC [4].
Thit k th nghim v x l s liu
Phng php qui hoch thc nghim
Xc nh hm lng ti u ca 3 yu t nh
hng glucose, cao nm men, FeSO4.7H2O
bng cch s dng qui hoch trc giao i
xng, mi yu t tin hnh ti 3 mc (-1, 0, +1)
nh trong bng 1. Qui hoch thc nghim a
ra bng ma trn thc nghim gm 20 th
nghim, trong : 16 th nghim ti tm (qui
hoch ton phn 24), 8 th nghim ti im sao
(2 th nghim cho mi bin) trong c 3 th

nghim lp ti tm, vi 1 hm mc tiu l th

tch kh trn tng th tch mi trng nui cy.
Cc th nghim cho nghin cu ti u c
tin hnh vi th tch dch mi trng 150 ml
trong bnh nui c th tch 150 ml iu kin k
kh, nhit 55oC vi t l tip ging (ging
ang pha sinh trng) 10% (v/v), tui ging
ang pha sinh trng.
X l s liu
X l s liu thc nghim bng phn mm
thng k Design-Expert 7.1 (Stat-Ease, Inc.,
Minneapolis, USA) phn tch cc h s hi
qui, b mt p ng v ti u ha vi thut ton
hm mong i.
KT QU V THO LUN

Chn min kho st

Khi nghin cu iu kin mi trng ph
hp cho to kh hydro ca chng
Thermoanaerobacterium aciditolerans Trau DAt
cho thy, qu trnh ny ph thuc vo nhiu yu
t nh t l tip ging, ngun cacbon, ngun
nit, hm lng FeSO4.7H2O, pH... Trong , ba
yu t glucose, cao nm men v st nh hng
mnh n kh nng sinh kh ca chng Trau DAt
(kt qu cha cng b). V vy, trong nghin cu
ny, chng ti chn min kho st ca ba yu t
ny tin hnh ti u iu kin nui cy ca
chng vi khun Trau DAt nh sau: glucose 8-12
g/l, cao nm men 2,5-3,5 g/l, FeSO4.7H2O 400600 mg/l vi hm mc tiu l th tch kh to ra
trn th tch dch nui cy. Mi tng quan gia
gi tr m ha v gi tr thc c ch ra bng 1
v phng trnh (1).

Bng 1. Gi tr m ha v gi tr thc nghim ca cc yu t thc nghim

Bin s
K hiu
n v
K hiu gi tr m ha
-1
0
+1
Glucose (A)
X1
g/l
8
10
12
Cao nm men (B)
X2
g/l
2,5
3
3,5
FeSO4.7H2O (C)
X3
mg/l
400
500
600
xi=(Xi-Xo)/Xi (1). Trong , xi l gi tr m ha ca yu t bin thin th i, Xi l gi tr tht ca yu t th i,
Xo l gi tr tht ca Xi ti im trung tm, Xi l bc nhy.

Thit lp m hnh
Gi tr m ha, kt qu thit k vi ma trn
k hoch thc nghim c trnh by bng 2.
470

Bng 2 gm 20 th nghim tng ng l 20 gi

tr khc nhau ca ba yu t glucose, cao nm
men, FeSO4.7H2O v th tch kh thu c

TP CH SINH HC 2013, 35(4): 469-476

tng ng vi cc gi tr ba yu t trn.
nh hng ca cc yu t glucose, cao nm
men, FeSO4.7H2O cng nh s tng tc gia
cc yu t n hm mc tiu (th tch kh thu
c) c tin hnh xy dng bi hm hi qui
bc 2 cho hm mc tiu (th tch kh do chng
Trau DAt to thnh) nh sau:

Yi=o + ki=0 ixi + iixi^2 + ijxixj (1)

Trong , Yi l hm mc tiu, o l h s t
do, i, ii, ij l cc vect tham s ca m hnh
c xc nh qua thc nghim. M hnh thng
k ch c ngha v c s dng sau khi tha
mn cc tiu chun thng k (Fisher).

Bng 2. Ma trn thc nghim vi 3 yu t glucose, cao nm men, FeSO4.7H2O v kt qu th nghim

Glucose
Cao nm men
FeSO4.7H2O
Th tch
K hiu
kh*
Nng
K hiu
Nng
K hiu
Nng
STT
mc
(ml/150ml
Glucose
mc tin
cao nm
mc tin FeSO4.7H2O
tin
dch nui)
(g/l)
hnh
men (g/l)
hnh
(mg/l)
hnh
1
6,64
-1
3,00
0
500,00
0
71,00
2
10,00
0
3,00
0
500,00
0
90,00
3
8,00
-1
2,50
-1
400,00
-1
65,68
4
8,00
-1
3,50
+1
400,00
-1
68,98
5
10,00
0
3,00
0
500,00
0
90,00
6
8,00
-1
3,50
+1
600,00
+1
80,00
7
12,00
+1
2,50
-1
600,00
+1
86,60
8
10,00
0
3,00
0
500,00
0
90,00
9
10,00
0
3,84
+1
500,00
0
89,00
10
10,00
0
3,00
0
331,82
-1
54,90
11
12,00
+1
3,50
+1
600,00
+1
88,60
12
10,00
0
3,00
0
500,00
0
90,05
13
8,00
-1
2,50
-1
600,00
+1
71,08
14
10,00
0
3,00
0
668,18
+1
73,00
15
13,36
+1
3,00
0
500,00
0
88,42
16
10,00
0
2,15
-1
500,00
0
85,00
17
12,00
+1
3,50
+1
400,00
-1
74,00
18
10,00
0
3,00
0
500,00
0
91,00
19
10,00
0
3,00
0
500,00
0
90,05
20
12,00
1
2,50
-1
400,00
-1
75,55
Phn tch s c ngha ca m hnh vi
thc nghim
Phn tch s ph hp ca m hnh v s c
ngha ca m hnh c nh gi qua phn tch
phn ANOVA (bng 3) v cc ch s tng
quan (bng 4). S c ngha ca cc h s hi qui
c kim nh bi chun F, vi cc gi tr p<
0,05 cho bit cc h s hi qui c ngha. Nh
vy, bng 3 cho thy gi tr Model-F-value l
994,2388 v m hnh hon ton c ngha
thng k vi tin cy 99,99% (p<0,0001). Vi
tt c cc yu t glucose, cao nm men,

FeSO4.7H2O v tng cp cc yu t ny u c
gi tr p<0,05 cho bit tng yu t ny cng nh
tng tc tng cp yu t u c ngha (bng 3).
iu ny c minh ha r hn khi quan st b
mt p ng hnh 1a, hnh 1b v hnh 1c.
Thm vo chun F cho s khng tng
thch-Lack of fit ca m hnh l 1,96
(p=0,2388), iu chng t m hnh hon
ton tng thch vi thc nghim. Kt qu thu
c cho thy, cc yu t glucose, cao nm
men, FeSO4.7H2O u c nh hng n qu
trnh to kh hydro sinh hc ca chng Trau
DAt. Kt qu nghin cu ca mt s tc gi
471

Nguyen Thi Yen, Lai Thuy Hien, Nguyen Thi Thu Huyen

khc cng ch ra nh hng cc yu t ny n

qu trnh to kh ca nhiu chng vi khun khc
nhau [6, 9].

v c ngha ca m hnh vi thc nghim. Kt

qu phn tch ANOVA cho thy gi tr R2 l
0,998 (R-Squared) bng 4 gn bng 1, chng
t gi tr th tch kh thu c t thc nghim
gn vi gi tr d on ca m hnh.

Bng 4 ch ra kt qu phn tch s ph hp

Bng 3. Kt qu phn tch ANOVA ti u qu trnh tng hp cc yu t

Tng bnh
phng
2128,591
341,647
27,55029
384,991
17,31661
10,64911
10,51111
192,4262
16,70669
1226,703
2,378806

Yu t
M hnh
A-glucose
B-cao men
C-FeSO4.7H2O
AB
AC
BC
A^2
B^2
C^2
Phn d
S khng
tng thch
Sai s thun
Tng tng quan

Bc
t do
9
1
1
1
1
1
1
1
1
1
10

Trung bnh
bnh phng
236,5101
341,647
27,55029
384,991
17,31661
10,64911
10,51111
192,4262
16,70669
1226,703
0,237881

5
5
19

0,315094
0,160667

1,575472
0,803333
2130,97

Gi tr F
994,2388
1436,212
115,8156
1618,421
72,7954
44,76663
44,18651
808,9193
70,23142
5156,803

Gi tr p
Prob > F
< 0,0001
< 0,0001
< 0,0001
< 0,0001
< 0,0001
< 0,0001
< 0,0001
< 0,0001
< 0,0001
< 0,0001

1,961169

0,2388

Tin cy

Khng tin
cy

Bng 4. Kt qu phn tch s ph hp ca m hnh vi thc nghim

Thng s
lch cun
Gi tr trung bnh
H s bin thin %
Tng bnh phng phn d
d on (PRESS)

Gi tr
0,48773
80,6455
0,604783
13,95849

Design-Expert Software

Design-Expert Software

Th tch kh
Design points above predicted value
Design points below predicted value
91

Th tch kh
Design points above predicted value
Design points below predicted value
91

101,5572

Th tch kh
Design points above predicted value
Design points below predicted value
91
92

93

54.9

84.5

Actual Factor
A: glucose = 10.00

80.75

77

54.9
87.5

X1 = A: glucose
X2 = C: FeSO4.7H2O

83

Actual Factor
B: cao men = 3.00

78.5

74

3.50

12.00
3.25
3.00

B: cao men

9.00
2.50 8.00

3.50
550.00

10.00
2.75

a. Glucose v cao nm men

600.00

3.25
500.00

A: glucose

80.5

74.25

68

600.00

11.00

86.75

T h tc h k h

X1 = B: cao men
X2 = C: FeSO4.7H2O

T h tch kh

88.25

T h tc h k h

Actual Factor
C: FeSO4.7H2O = 500.00

Gi tr
0,998884
0,997879
0,99345

Design-Expert Software

92

54.9
X1 = A: glucose
X2 = B: cao men

Thng s
R2
R2 hiu chnh
R2 d on
chnh xc ph hp
(Adeq Precision)

C: FeSO4.7H2O

2.75
400.00 2.50

12.00
550.00

3.00
450.00

11.00
500.00

B: cao men

b. FeSO4.7H2O v cao nm men

C: FeSO4.7H2O

10.00
450.00

9.00

A: glucose

400.00 8.00

c. FeSO4. 7H2O v glucose

Hnh 1. B mt p ng ca tng cp yu t nh hng

n qu trnh to kh hydro chng Trau DAt
T cc gi tr phn tch c ngha trn, gi
tr hm mong i c phn mm DX7 a ra
c biu din theo phng trnh c th sau:
472

Y = -311,574 + 22,300X1 + 31,931X2 + 0,849

X3 -1,471X1X2 + 0,006X1X3 + 0,023X2X3 0,913X12 - 4, 307X22 - 0,0009X32 (2).

TP CH SINH HC 2013, 35(4): 469-476

Trong , Y l th tch kh mong i; X1,

X2, X3 l gi tr hm lng glucose, cao nm
men v FeSO4.7H2O.
Ngoi ra, 39 gii php ti u vi hm lng
3 bin xc nh l glucose, cao nm men v

FeSO4.7H2O t s dng thut ton hm mong

i bng phng php p ng b mt cng
c a ra, kt hp vi phng trnh hm mong
i tm ra, th tch kh tnh c tng ng
vi 3 bin xc nh c trnh by bng 5.

Bng 5. Cc gii php ti u vi hm lng 3 bin xc nh v gi tr hm mong i ti u

Th tch kh
Glucose
Cao nm men
FeSO4.7H2O
STT
(mg/l)
(ml/150 ml dch nui)
(g/l)
(g/l)
1
8,00
3,50
400,00
87,75
2
10,00
3,00
500,00
139,03
3
12,00
3,50
400,00
164,45
4
12,00
3,50
600,00
163,62
5
8,00
3,50
600,00
82,31
6
8,00
2,50
600,00
87,99
12,00
600,00
7
2,50
175,19
8
8,00
2,50
400,00
93,43
12,00
400,00
9
2,50
176,02
10
9,72
2,97
471,42
133,28
11
9,09
3,00
452,00
119,28
12
11,82
3,03
545,18
173,87
13
8,46
2,76
534,64
106,82
14
11,44
3,11
488,28
167,26
15
8,92
3,44
434,14
110,49
16
8,07
2,98
572,06
92,91
17
10,06
2,87
471,22
141,02
18
9,04
3,41
482,98
115,87
19
9,32
2,93
561,78
120,90
20
9,89
2,57
410,80
133,82
21
10,41
2,96
578,82
141,01
22
9,79
2,57
462,38
137,02
23
10,85
2,80
466,30
157,72
24
11,07
3,18
487,16
158,94
25
10,45
2,98
425,70
144,49
26
10,37
2,52
512,50
149,73
27
11,75
3,36
489,46
170,25
28
9,83
2,62
544,02
135,70
29
10,27
2,87
524,98
144,70
30
8,52
3,03
417,32
104,30
31
8,25
3,47
599,82
87,72
32
9,30
3,10
413,74
118,88
33
8,21
3,16
511,42
100,92
34
11,99
3,42
401,62
165,84
35
11,56
2,68
566,36
169,56
36
10,63
2,80
461,00
152,75
37
11,86
2,54
409,82
174,53
38
9,90
3,39
416,00
127,90
39
8,24
2,89
482,29
103,89
473

Nguyen Thi Yen, Lai Thuy Hien, Nguyen Thi Thu Huyen

Bng kt qu ti u cho thy vi 3 cp gi

tr bin glucose, cao nm men, FeSO4.7H2O
khc nhau cc gi tr hm mong i thu c l
khc nhau. Bng 5 cho thy, 5 t hp thnh
phn glucose, cao nm men, FeSO4.7H2O ti
cc im c s th t l 7, 9, 12, 27, 37 cho th
tch kh mong i cao hn c v tng ng
nhau. Tin hnh kim tra tnh ng n ca m
hnh ti u, tin hnh cc th nghim kim

chng ti 5 im ti u m hnh a ra bng 5

t chn cp bin cho kt qu hm mong i
ti u nht (bng 6).
T kt qu kim tra thc nghim bng 6,
gi tr ti u cho 3 bin c xc nh nh sau:
glucose 12 g/l, cao nm men 2,5 g/l,
FeSO4.7H2O 400 mg/l, th tch kh thu c l
176 ml /150 ml dch nui cy.

Bng 6. Kt qu kim tra th tch kh thu c t m hnh v thc t

STT
m
hnh
7
9
12
27
37

Glucose
(g/l)

Cao nm
men (g/l)

FeSO4.7H2O
(mg/l)

12,00
12,00
11,82
11,75
11,86

2,50
2,50
3,03
3,36
2,54

600,00
400,00
545,18
489,46
409,82

Hnh 2. Thnh phn kh do chng

Thermoanaerobacterium aciditolerans Trau
DAt to ra (t tri qua phi ph cao nht l ph
H2, tip theo l ph H2S, cui cng l ph CO2)
trong iu kin ti u
Ln men qui m 600 ml trong iu kin ti
u xc nh
Sau khi xc nh c hm lng ti u
cho 3 yu t, chng ti tin hnh ln men th
tch dch nui cy 600 ml trong iu kin ti u
xc nh th tch kh thu c cng nh phn
tch thnh phn kh do chng vi khun
Thermoanaerobacterium aciditolerans Trau
DAt to ra. Kt qu phn tch thnh phn kh
hnh 2 cho thy, thnh phn kh hydro to ra
chim 77,2% tng th tch kh, t 420 ml kh

474

Th tch kh theo
thut ton (ml/150ml
dch nui cy)
175,19
176,02
173,87
170,25
174,53

Th tch kh theo
thc t (ml/150ml
dch nui cy)
175,00
176,02
172,60
169,00
174,17

hydro/600 ml dch nui trong iu kin ti u.

Th tch kh hydro to ra cao gn gp 5 ln th
tch kh hydro trc khi ti u ca chng vi
khun ny (th tch kh hydro thu c khi cha
ti u l 85 ml/600 ml dch nui cy-kt qu
cha cng b). Kt qu ny mt ln na khng
nh vic s dng phng php ton hc qui
hoch thc nghim hon ton c ngha v
mang li hiu qu cao trong nghin cu nh
hng ca cc yu t glucose, cao nm men,
FeSO4.7H2O n qu trnh to kh hydro sinh
hc
ca
chng
vi
khun
Thermoanaerobacterium aciditolerans Trau
DAt.
KT LUN

S dng phng php ton hc qui hoch

ha thc nghim bng p ng b mt xc
nh c hm lng ti u cho mi trng ln
men to kh hydro sinh hc ca chng
Thermoanaerobacterium aciditolerans Trau
DAt vi 3 yu t glucose l 12 g/l, cao nm men
l 2,5 g/l, FeSO4.7H2O 400 mg/l. C 3 yu t
cng nh s tng tc gia cc yu t ny u
c nh hng n qu trnh to kh hydro ca vi
khun ny. Ln men th tch 600 ml dch nui
cy thu c lng kh hydro l 420 ml, cao
gp 5 ln th tch kh hydro khi cha ti u

TP CH SINH HC 2013, 35(4): 469-476

thnh phn ba yu t glucose, cao nm men,

FeSO4.7H2O, iu ny chng t s dng
phng php qui hoch ha thc bng p ng
b mt ph hp cho nghin cu ti u to kh
hydro ca chng vi khun Trau DAt.
Li cm n: Cng trnh c thc hin bi s
ti tr ca Vin Hn lm Khoa hc v Cng
ngh Vit Nam (VAST 05.02/11-12) v s hp
tc ca Vin Nghin cu v pht trin ng dng
cc hp cht thin nhin (trng i hc Bch
khoa H Ni).
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Nguyen Thi Yen, Lai Thuy Hien, Nguyen Thi Thu Huyen

OPTIMIZATION OF MEDIUM COMPONENTS FOR HYROGEN PRODUCTION

OF ANAEROBIC BACTERIAL STRAIN Thermoanaerobacterium aciditolerans Trau
DAt ISOLATED FROM VIETNAM BY USING RESPONSE SURFACE
METHODOLOGY
Nguyen Thi Yen1, Lai Thuy Hien1, Nguyen Thi Thu Huyen1,2
1

Institute of Biotechnology, VAST

Nguyen Tat Thanh University, Ho Chi Minh city

SUMMARY

The 21st century is era of renewable energy in which biohydrogen is the most potential candidate that may
replace the fossil fuel. Many countries pay more attention to biohydrogen since it can produce by bacteria via
fermentation and gas-biosynthesis has many advantages. Bacterial strain Thermoanaerobacterium aciditolerans
Trau DAt isolated from Vietnam is capable of biohydrogen production in anaerobic condition. The gas volume
as well as the gas components produced by the strain Trau DAt depends on nutrient and fermentation conditions.
Hydrogen production of strain Trau DAt was highly affected by glucose, yeast extract and FeSO4.7H2O. By
using response surface methodology (RSM) with central composite design, the mutual effects of these factors on
hydrogen production of strain Trau DAt were studied. RSM analysis showed that the highest hydrogen
production potential (176.02 ml/150 ml culture medium) was obtained under the condition of glucose 12 g/l,
yeast extract 2.5 g/l, FeSO4.7H2O 400 mg/l. Under optimized condition, the maximum H2 volume of 420 ml/600
ml culture medium was obtained by dark fermentation.
Keywords: Biohydrogen, RSM, bacteria, Vietnam.

Ngy nhn bi: 25-6-2013

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