TAP CHI STNH HOC 2013, 35(4): 469^76
T O I U t J H O A T H A N H P H A N M O I T R U ' O N G T A O K H I H Y D R O S I N H H O C C U A C H U N G V I K H U A N K I K H I Thermoanaerobacterium aciditolerans T r a u D a t P H A N L A P T A I V I E T N A M B A N G P H U O N G P H A P D A P U N G B E M A T ( R S M )
Nguyen T h j Y e n ' , L a i Thiiy H i e n ' , Nguyen Thi T h u Huyen'"^*
'Vien Cdng nghe sinh hoc, Vien Han lam KH & CN Viet Nam, *[email protected]
^Trudng Dai hoc Nguyen Tat Thanh, thanh ph6 Ho Chi Minh
1 OM TAT: The ky 21 la ky nguyen ciia nguon nang lugng tai tao, trong do nang luong hydro smh hoc la lira chon sang gia co the thay the nguon nhien lieu hoa thach dang ngay cang can kiel. Hydro sinh hoc dang dugc nhieu nuoc quan tam, boi no co nhieu iru diem va duoc tao ra bang con duong len men vi sinh vat Chung vi khuan Trau DAt phan lap 6 Viet Nam co kha nang tao khi H2 sinh hoc trong dieu kien ten men ki khi. The lich khi cung nhu thanh phan khi lao ra ciia chimg vi khuan nay phu thuoc nhieu vao cac yeu to dmh duong va dieu kien moi truong nuoi cay. Khi nghien cihi anh hirong cua timg yeu to dinh du&ng trong moi truang nuoi cay cho thay glucose, cao nam men, FeS04.7H20 co tac dong manh den qua trinh tao khi H2 ciia chiing vi khuan Trau DAt Sir dung phucmg phap qui hoach hoa thuc nghiem bang dap iing be mat dua tren kieu tam phuc hop vdi 3 bicn la glucose, cao nam men va FeS04 7H2O, da xac dinh duoc su tuong tic giua cac yeu to neu tren cung tac dong den the tich khi hydro sinh hoc do chiing Trau DAt tao ra Phuong phap qui hoach thuc nghiem cung xac dinh dugc ham lugng toi uu ciia 3 yeu to nay cho qua trinh lao khi hydro la glucose 12 g/l; cao men 2,5 g/1; FeSOj 7H2O 400 mg/l, Thi ti'ch khi thu duoc khi len men chiing Trau DAt VOI cac thong so toi uu noi tren la 176,02 ml/150ml moi trudng Len men theo me vdi the tich 600 ml moi Injong trong dieu kien toi uu thu duoc 420 ml khi hydro.
Tit khoa: Hydrogen smh hpc, RSM, vi khuan, Vict Nam.
MO DAU
Hydro sinh hgc duoc coi nliu ngu6n nang lugng Ihay the trong tuong lai do kbi dot chay sinh ra nang lucmg lan, khong Iao ra kbi CO;, vi vay, khong gay 6 nhiem moi trub'ng. Nghien cuu san xuat hydro sinh hgc da duoc nhieu quoc gia Iren the gigi quan tam, Irong do cb qua trinh san xuat khi hydro sinh hgc tir cac vi khuan ki khi [I, 6. 10]. Cac k i t qua nghien cihi qua trinh len men Ihu khi hydro tir vi khuan ki khi cho Ihay qua trinb nay bi anh huong bai nhieu yeu to dinh duong cung nhu yeu to moi truong. Do do, 161 uu hoa cac thanh phan moi truong nuoi cay nham nang cao kha nang tao khi hydro ciia cac chiing vi khuan rat quan trgng [ I , 2, 3, 6, I I ] . Gan day, nghien ciiu toi iru cac yeu to moi truong de len men tao kbi hydro su dung phuong phap qui hoach thuc nghiem bang dap img be m3l dugc nghien cuu nhieu [7, 8, 12].
Su dung phuong phap loan hgc qui hoach thuc nghigm cho phep xac dmh dugc dieu kien toi uu ciia da yeu t6 mgt each chinh xac va miic do anh huong ciia cac yeu to den qua trinh tao khi hydro Irgng ciing mgt thiri diem Them vao do.
phugng phap nay con cho Ibay vai Iro lac dgng qua lai giQa cac yeu to trong ciing mpt thai gian nu6i cay vcri so lan thi nghiem it. danb gia dugc sai s6 trong moi lan thuc nghiem theo cac tieu chuan thong ke, xem xet anh huang cua cac yeu t6 vol muc do tin cay can thiet, tiet kiem Ihgi gian va chi phi ma van dem lai hieu qua mong muon Bang viec su dung phuong phap qui hoach thuc nghiem, nhieu tac gia da cbi ra su luong tac qua lai giira cac yen to m6i truang c6 tac dgng den qua trinh tao khi hydro. Dong thoi, su dung phuang phap nay cung da xac dinh duoc nong do toi uu cho cac yeu to dinh duong va moi truong cho qua Irinh len men Iao khi hydro cua cac chung vi khuan [7, 8, 9, 12]
Trong cong trinh nay, chiing t6i trinh bay ket qua l6i uu hoa ba yeu to glucose, cao nam men, FeS04.7H20 trong moi truong len men Iao khi hydro sinb hgc tii chung vi khuan ki khi Thermoanaerobacterium aciditolerans Trau DAt bing phuong pbap qui hoach hoa Ihuc nghiem dap img be mat (Response Surface Methodology-RSM) va ket qua phan tich Ihanh phan khi khi len men 6 the tich 600 ml dich nuoi cay trong dieu kien t6i uu tim dugc.
Nguven Thi Yen. Lat Thuy Hien. Nguyen Tlu Thu Huyei
VAT LIEU VA PHU'O'NG PHAP NGHIEN CUlT C h u n g giong va moi t r u o n g nuoi cay
Chung VI khuan ki kbi Thermoanaerobacterium aciditolerans Trau DAt dugc pban lap tren men truong chon ioc N M V tir mau pban Irau D6ng Anh, Ha Noi [4]
M6i truong nu6i cay chiing vi khuan Trau DAt la moi truang N M V [4],
P h u o n g p h a p
Nu6i cay vi khuan ki khi tren moi tnrong chon loc NMV,
Su dung phuong phap qui hoach hoa thuc nghiem bang dap img be mat de thiet ke ma tran thuc nghiem [4],
Xac dinh the tich khi hydro bang phuong phap the cli6 nuoc (water displacement method) [4].
Phan tich chat luong va ham lugng khi hydro bang may sac ki khi GC-TCD (Thermo Trace GC-Tbermo Electro-USA) vai phuong phap thtr EDC VI-003 GC [4],
Thiet ke thi nghiem va xir ly so lieu Phwcmg phap qui hoach ihyc nghiem
Xac dinh bam lugng toi uu cua 3 yeu to anh huong glucose, cao nam men, FeS04 7HiO bang each sir dung qui boacli true giao d6i xirng, m6i yeu to lien banh tai 3 muc (-1, 0, +1) nhu trong bang 1 Qui hoach tliuc ngliiem dua ra bang ma Iran thuc nghiem gom 20 thi nghiem, trong do. 16 thi nghiem tai Iam (qui hoach loan phan 2"), 8 thi nghiem tai diem sao (2 thi nghiem cho moi bien) trong do co 3 thi
nghiem lap tai tam, vai 1 ham muc tieu !a the tich khi tren tong the tich m6i truong nuoi cay
Cac thi nghiem cho nghien ciru l6i un duoc ti8n hanh vm the tich dich ITI6I truang 150 ml trong binh nuoi co the tich 150 ml a dieu kien ki khi, nhiet do 5 5 ^ voi ty le tigp giong (giong dang a pha smh truang) 10% (v/vj, moi giong dang a pba sinh t r u a n g
Xif ly so lieu
X u li so lieu thuc nghiem bang phan mem thong ke Design-Expert 7 1 (Stat-Ease, Inc., Minneapolis, USA) de phan ticb cac he so hoi qui, be mat dap iing va toi uu hoa vgi thual loan ham mong dgi.
KET QUA VA THAO LUAN C h g n mien k h a o sat
Khi nghien ciui dieu kien moi tmong phii hop cho tao khi hydro ciia chiing Thcrmoanaernhaclerium aciditolerans Trau DAl clio thay, qua trinh nay pbu thugc vao nhieu yeu to nhu ty le Hep gi6ng, nguon cacbon, nguon nito, ham luang FeS04.7H20, pH .. Trong do, ba yeu t6 glucose, cao nam men va sat anh huong manh den kba nang smh khi cua chting Trau DAt (ket qua chua cong bo). Vi vay, Irong ngbien cim nay, chiing loi chgn mien khao sat cua ba yeu to nay de tien banh toi un dieu kien nu6i cay cua chung VI khuan Trau DAt nhu sau glucose 8-12 g/l, cao x\km men 2,5-3,5 g/1, FeSOj 7 H : 0 400- 600 mg/l vdl ham muc tieu la the tich kbi Iao ra tren the ticb dich nu6i cay. Moi luong quan giiia gia tri ma hoa va gia tn thuc dugc cbi ra 6 bang I va phu'ang trinh (1),
Bdng I Gia tri ma boa va gia tri thuc nghiem cua cac yeu t6 Ihuc nghie Bien .s6
Glucose (A) Cao nam men (B) FCSO4 7H2O (C)
Ky hieu X|
X . X ,
Don Vj R/l R''l mg/l
Ky hicu gia tn ma hoa -1
8 2,5 400
0 10 3 500
+1 12 3,5 600 x=(X,-Xi,)/AX, (1) Trong do. x, la gia tn ma hoa ciia yeu to bicn thicn thir 1, X, la gia Ui chat cua yeu to thir 1, X„ la gia tn that cua Xi tai diem tmng tam, AX, la budc nhay
Thiet lap mo hinh Bang 2 gom 20 thi ngbiem t u a n g irng la 20 gia Gia tri ma boa, k i t qua tliifil k^ vai ma tran ^n khac n l i a u c i i a ba y6u 16 glucose, cao nam kc hoach thuc nghiem d u g c trinli bay o bang 2 FcSOj 7 H ; 0 va 'cb kill Ihu dugc
TAP CHI SINH HOC 2013, 35(4) 469-476 mong ung voi cac gia tn ba yeu to tren.
.\nh buong ciia cac yeu to glucose, cao nam men. FeSOj 7H2O ciing nhu su tuong tac giua cac yeu to den ham muc lieu (the tich khi thu duoc) duac tien hanh xay dung bai bam hoi qui hac 2 cho ham muc Ueu (the tich khi do chiing Trau DAt lao thanh) nhu sau'
Y,=p„ + I ' , ^ p,x, + I p„x,-2 + X p„x,x, (1)
Trong do, Yi la ham muc lieu, po la he so tu do, PI. PH, pij la cac vecto tham so ciia mo hinh duac xac dmh qua thuc nghiem. Mo hinh th6ng ke chl cb y nghTa va duoc sir dung sau khi thoa man cac lieu chuan thong ke (Fisher) Bang 2STT
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20
\1a Iran thuc nghiem va Glucose Nong 66 Glucose (E") 6,64 10,00 8,00 8,00 10,00 8,00 12,00 10,00 10,00 10,00 12,00 10,00 8,00 10,00 13,36 10.00 12,00 10,00 10,00 12,00
Ky hieu muc tien hanh
-1 0 -1 -1 0 -1 + 1 0 0 0 +1 0 -1 0 + 1
0 H 0 0 1
3 yeu to glucose, cao nam men, F e S 0 j - 7 H ; 0 va ke Cao nam men
Nong do cao nam men (g/1) 3,00 3,00 2,50 3,50 3,00 3,50 2,50 3,00 3,84 3,00 3,50 3,00 2,50 3,00 3,00 2,15 3,50 3.00 3,00 2,50
Ky hieu miJc tien hanh
0 0
•1 + 1
0 + 1 -1 0 +1 0 +1 0
FeSOj 7 H , 0 Nong dp FeSO^ 7 H . 0
(mg'l) 500.00 500,00 400,00 400,00 500,00 600.00 600.00 500,00 500,00 331,82 600,00 500,00 -1 ' 600,00 0
0 -1 + 1
0 0
•1
668,18 500,00 500,00 400,00 500,00 500,00 400,00
Ky hieu mirc ti^n hinh 0 0 -1 -1 0 + 1 + 1 0 0 -1 + 1 0 + 1 H 0 0 -1 0 0 -1
qua thi nghiem The tich
khi*
(ml'!50ml dich nuoi) 71.00 90.00 65.68 68,98 90,00 80.00 86.60 90,00 89,00 54,90 88,60 90,05 71,08 73,00 i 88,42 85,00 74,00 91,00 90,05 75,55 Phan tich sir c6 nghia ciia m o hinh voi
Ihirc nghiem
Phan tich su phii hop cua mo hinh va su c6 nghTa cua m6 hinh dugc danh gia qua phan tich phSn ANOVA (bang 3) va cac chi so tuong quan (bang 4) Su co ngliTa cua cac be s6 hoi qui dugc kicm dmh boi chuan F, vai cac gia in p<
0,05 cho bicl cac he so hoi qui co nghTa Nhu vay. bang 3 cho thay gia Iri "Modcl-F-valuc" la 994.2388 va m6 hinh hoan loan co y nghTa thong kc voi d6 tin cay 9 9 , 9 9 % (p<0,000l), Voi tat ca cac yeu 16 glucose, cao nam men,
FeSOj 7 H ; 0 va timg cap cac yeu 16 nay deu co gia In p<0.05 cho biel tiing yeu to nay ciing nhu tuong tac timg cap yeu to deu co nghTa (bang 3) DICU nay duoc mmh hoa ro hon khi quan sal be mat dap irng a binh la, hinh lb va hinh Ic Them vao do chuan F cbo "su khong tuong tliich-Lack of fit" ciia mo binh la 1.96 (p=0,2388), dieu do chung lb mo hinh hoan loan luong thich voi Ihuc nghiem Kcl qua thu dugc cho thay, cac yeu to glucose, cao nam men. FcS04,7H;0 deu co anh huong den qua Irinh Iao khi hydro sinh hgc cua chung Trau DAt Kcl qua nghien cuu cua mot so Iac gia
Nguyen Thi Yen, Lai Thuy Hien. Nguyen Tin Thu Huyen khac cung chi ra anb b u o n g cac yen to nay den
qua trinh tao khi cua nhieu cbung vi khuan khac nhau [6, 9],
Bang 4 chi ra ket qua pban tich su phii hgp
va CO nghTa cua m o hinh vai tbyrc ngbiem Ket qua phan tich A N O V A cho t h i y gia trj R^ la 0,998 (R-Squared) 6 bang 4 gan biing 1, chung to gia tri thg tich khi thu d u g c tir thuc nghiem gan vai gia tri d u doan ciia m o hinh.
Bdng 3. Ket qua phan ticb A N O V A toi uu qua trinh tong bgp cac yeu t6 Yeu to
Mo hinh A-glucose B-cao men C - F e S 0 4 . 7 H , 0 AB AC BC A»2 B"2 C-^2 Phan du Sir khong tuong thich Sai so thuan Tong tuong quan
Tong binh p h u o n g 2128,591
341,647 27,55029 384,991 17,31661 10,64911 10,51111 192,4262 16,70669 1226,703 2,378806 1,575472 0,803333 2130,97
Bac tu do
9 1 1 1 1 1 1 1 1 1 10 5 5 19
Tioing binh binh phuong
236,5101 341,647 27,55029 384,991 17,31661 10,64911 10,51111 192,4262 16,70669 1226,703 0,237881 0,315094 0,160667
Gia tri F
994,2388 1436,212 115,8156 1618,421 72,7954 44,76663 44,18651 808,9193 70,23142 5156,8031,961169 Gta tri p Prob > F
< 0,0001
< 0,0001
< 0,0001
< 0,0001
< 0,0001
< 0,0001
< 0,0001
< 0,0001
< 0,0001
< 0,0001
0,2388
Tin cay
Khong tin cay
Bang 4 Ket qua phan tich su phu hgp ciia mo hinh voi thuc nghiem Thong so
D6 lech cuan Gia In Irung binh He so bien thien % T6ng binh p h u a n g phan du du doan (PRESS)
Gia Iri 0,48773 80,6455 0.604783 13,95849
Tb6ng so R '
R" bieu chinh R" du doan Do chinh xac phii hgp (Adeq Precision)
Gia tn 0,998884 0,997879 0,99345 101,5572
a. Glucose va cao nam men b. FeS04.7H20 va cao nam men c. FeS04. 7H2O va glucose Hinh 1. Be mat dap ling ciia timg cap yeu to anh h u o n g
den qua trinb tao khi hydro a cluing Trau D A t Tir cac gia tri phan tich co ngbia a tren, gia
In bam mong dgi dugc phan mem DX7 dua ra dugc bieu dien theo phuong trinh cu the sau:
Y = -311,574 + 22,300X, + 31,931X2 + 0,84f Xj -1,471X1X3 + 0,006X1X3 + 0,023X2X3 0 , 9 1 3 X r - 4, 307X2^ - 0,0009X3' (2).
TAP CHI SINH HOC 2013, 35(4): 469^76 Trong do, Y la the tich khi mong dgi; Xj,
X2, X J la gia tri ham lugng glucose, cao nam men va FeSOj.7H20.
Ngoai ra, 39 giai phap toi uu voi ham lugng 3 bien xac dinh la glucose, cao nam men va
FeS04.7H20 tu su dung Ihuat toan ham mong dgi bang phucmg phap dap iing be mat ciing dugc dua ra, ket hgp vcii phuong trinh ham mong dgi da tim ra, the tich khi tinh dugc tucmg ung vdi 3 bien xac dinh dugc trinh bay a bang 5.
Bang 5.
STT 1 2 3 4 5, 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 2S 29 30 11 t2 r , 14 IS M' 37 38 19
Cac giai phap toi uu vai ham Itrcmg Glucose
8,00 10,00 12,00 12,09 8,00 8,00 12,00
8,00 12,00 9,72 9,09 11,82
8,46 11,44
8,92 8,07 10,06 9,04 9,32 9,89 10,41 9,79 10,85 11,07 10,45 10,37 11,75 9,83 10,27 8,52 8.25 9.30 8,21 11.99 11,56 10.63 11,86 9,90 8,24
Cao nam men (g/l) 3,50 3,00 3,50 3,50 3,50 2,50 2,50 2,50 2,50 2,97 3,00 3,03 2,76 3,11 3,44 2,98 2,87 3,41 2,93 2,57 2,96 2,57 2,80 3,18 2,98 2,52 3,36 2.62 2.87 3.01 3,47 3,10 3,16 3,42 2,68 2,80 2,54 3,39 2,89
3 bien xac dinh va gia FcSOj.VHiO
(mg/l) 400,00 500,00 400,00 600,00 600,00 600,00 600,00 400,00 400,00 471,42 452,00 545,18 534,64 488,28 434,14 572,06 471,22 482,98 561,78 410,80 578,82 462,38 466,30 487,16 425,70 512,50 489,46 544,02 524,98 417,32 599,82 413,74 511,42 401,62 566,36 461,00 409,82 416,00 482,29
tri ham mong doi toi uu The tich khi (ml/150 ml dich nuoi)
87,75 139,03 164,45 163,62 82,31 87,99 175,19 93,43 176,02 133,28 119,28 173,87 106,82 167,26 110,49 92,91 141,02 115,87 120,90 133,82 141,01 137,02 157,72 158,94 144,49 149,73 170,25 135,70 144,70 104,30 87,72 118,88 100,92 165,84 169,56 152,75 174,53 127,90 103,89
Nguyen Thi Yen. Lai Thuy Hien. Nguyen Thi Thu Huyen Bang ket qua toi uu cho thay VO'i 3 cap gia
tri bien glucose, cao nam men, FeS04.7H20 kbac nhau cac gia tri ham mong dgi thu d u a c la kbac nhau. Bang 5 cbo thay, 5 to bgp thanh phan glucose, cao nam men, FeS04.7H20 tai cac diem co s6 tbu tu la 7, 9, 12, 27, 37 cho the Iich kbi mong dgi cao hon ca va tuang d u a n g nhau. Tien banh kiem tra tinh diing dan ciia m6 hinh Ioi uu, tien hanh cac thi nghiem kiem
chiing tai 5 diem toi u u m o hinh d u a ra a bang 5 tir do chgn cap bien cho ket qua ham mong dgi t6i uu nhat (bang 6).
Tir ket qua kiem tra tbyc nghiem a bang 6, gia tri t6i uu cho 3 bien d u g c xac djnh n b u sau:
glucgse 12 g/l, cao nam men 2,5 g/l, F e S 0 4 . 7 H 2 0 400 mg/l, I h l tich khi thu dugc la 176 ml /150 ml dich nu6i cay.
Bdng 6. Ket qua kiem tra the tich khi thu dugc tii m6 binh va thuc te
STT
mo hinh 7 9 12 27 37
Glucose (g/1) 12,00 12,00 11,82 11,75 11,86
Cao nam men (g/1) 2,50 2,50 3,03 3,36 2,54
FeS04,7H,O (mg/l) 600,00 400,00 545,18 489,46 409,82
The tich khi theo thuat toan (ml/150ml
dich nuoi cay) 175,19 176,02 173,87 170,25 174,53
The tich khi theo thtrc te (ml/150ml dich nuoi cay)
175,00 176,02 172,60 169,00 174,17
Hinh 2. Thanh phan khi do chiing Thermoanaerobacterium aciditolerans Trau DAt tao ra (tir trai qua phai pho cao nhat la pho Hi. Hep Iheo la ph6 HiS, cuoi ciing la ph6 CO2)
trong dieu kien toi uu Len men qui m o 600 ml t r o n g d i l u k i l n toi un da xac dinh
Sau khi da xac dinh dugc ham luong tai uu cho 3 yeu to, chiing toi tien hanh len men o the Iich dich nuoi cay 600 ml Irong diSu kien t6i im de xac dinh the Iich kbi tbu d u a c ciing n b u phan tich Ihanh phan khi do chiing vi khuSn Thermoanaerobacterium aciditolerans Trau DAt tao ra Ket qua pban lich thanh p h i n khi a hinh 2 cho thay, thanh phan khi hydro tao ra chiem 77,2% tfing the tich khi, dat 420 ml khi
hydro/600 ml dich nu6i trong dieu kien toi uu.
The tich kbi hydro tao ra cao gan gap 5 lan the tich khi hydro truoc kbi toi uu cua chiing vi khuan nay (the tich khi hydro tbu duoc khi chua toi uu la 85 ml/600 ml dicb nu6i cay-ket qua chua cong bo). Ket qua nay mpl lan nija khang dinb viec sii dung p h u a n g phap loan hgc qui hoach thuc nghiem boan toan co y nghTa va mang lai bieu qua cao trong nghien cuu anh huong ciia cac yeu to glucose, cao nam men, FeS04.7H20 den qua trinh tao kbi hydro sinh bgc ciia chung vi khuan Thermoanaerobacterium aciditolerans Trau DAt.
KET LUAN
Sii dung p h u a n g phap toan hpc qui hoach hoa thuc nghiem bang dap ting be mat da xac dinh d u g c ham lugng Ioi uu cho moi Irucmg len men tao khi hydro sinh hgc cua chiing Thermoanaerobacterium aciditolerans Trau DAt vdl 3 yeu 16 glucase la 12 g/l, cao nam men la 2,5 g/l, F e S 0 4 . 7 H 2 0 400 mg/l. Ca 3 y^u _t6 Cling n b u sir tuong tac giiia cac ygu t6 nay deu CO anh h u a n g den qua trinh tao khi hydro cua vi khuan nay. Len men a the tich 600 ml djch nuoi cay thu d u g c luong khi hydro la 420 ml, cao gap 5 lan the tich khi hydro khi chua toi mi
TAP CHI SINH HOC 2013, 35(4): 469-476 thanh phan ba yen to glucose, cao nam men,
FeS04.7H20, dieu nay chung to su dung phuong phap qui hoach hoa thuc bang dap iing be mat phii hgp cho nghien ciiu Ioi uu tao khi hydro ciia chiing vi khuan Trau DAt, Loi cdm cm Cong Irinh dugc thuc hien bcri su tai trg cua Vien Han lam Khoa hoc va Cong nghe Viet Nam (VAST 05.02/11-12) va su bgp tac ciia Vien Nghien ciiu va phat trien img dung cac hcrp chat thien nhien (truang Dai hgc Bach khoa Ha Ngi).
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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 Yen', Lai Thuy Hien", Nguyen Thi Thu Huyen"'^
'institute ofBiotechnology, VAST
^Nguyen Tat Thanh University, Ho Chi Minh city
The 21st century is era of renewable energy in which biohydrogen is the most potential candidate that may replace the fossil fiiel. Many countnes pay more attention to biohydrogen since it can produce by bactena via femientation and gas-biosynthesis has many advantages, Bactenal strain Thermoanaerobaclermm 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 femientation conditions.
Hydrogen production of strain Trau DAt was highly affected by glucose, yeast extract and FeSO^ 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 condiUon of glucose 12 g/l, yeast extract 2,5 g/1. FcSOj THjO 400 mg/l. Under optimized condition, the maximum Hj volume of 420 ml/600 ml culture medium was obtained by dark femientation.
Keywords Biohydrogen, RSM, bactena, Vietnam.
Ngdy nhdn bdi. 25-6-2013
