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cng lun vn tt nghip

S GIO DC V O TO TRNG I HC KHOA HC T NHIN H QG TP.HCM KHOA SINH HC CNG KHA LUN TT NGHIP

ST S TNG TRNG V S TCH LY LIPID TRONG T BO TO NC NGT TRN MI TRNG LNG.


TI: KHO Sinh vin: NGUYN TH PHNG THANH Mssv: 0818290 Ngnh: Cng ngh sinh hc

1.M U. 1.1 Tng quan v nhin liu sinh hc. Trong bi cnh th gii ngy nay, vic m bo ngun nng lng di hn thay th nng lng ho thch ang ngy cng tr nn cp thit, nht l khi du m ang cn dn v tr nn t . Hin tng tri t ang nng dn ln v gia tng pht thi kh ccbonic -kh gy ra hiu ng nh knh i hi pht trin cc ngun nng lng mi. Nhin liu sinh hc (NLSH Biofuel) l nhin liu ti to (renewable fuels) c sn xut t nguyn liu sinh hc (sinh khi), c dng kh v lng, c dng trong vn ti. NLSH dng lng bao gm ethanol sinh hc (bioethanol), diesel sinh hc (biodiesel), FischerTropsch fuel, dng kh gm hydro sinh hc (biohydro) v metan sinh hc (biometan). Th h NLSH u tin c sn xut ch yu t cc loi cy lng thc, thc phm nh ng, sn, ma, u nnh, c, ht ci, vv tuy lm gim ng k kh CO2 pht thi so vi nhin liu ha thch, nhng khng thc s pht trin bn vng do nguyn liu cho NLSH li l ngun lng thc cho con ngi v gia sc. Gia tng s dng ngun nguyn liu cho NLSH thuc loi nu trn s lm gia tng gi c cc loi cy trng ny
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cng lun vn tt nghip

v lm gia tng bt n lng thc. Ngoi ra, pht trin nguyn liu cho NLSH, qu t trng trn th gii s khng p ng (Chisti, 2008a). Vn tm kim v pht trin cc loi nhin liu thay th v ci tin hn v mt cng ngh c tip tc t ra. Th h NLSH th hai c pht trin ch yu da vo lignocellulosic ca cc loi cy khng phi l cy lng thc v c th trng cc vng t cn ci, hoang ha (v d nh cy c ngt (sweetgrass), cy cc ro (jatropha). Mc d nguyn liu th cho sn xut NLSH th h 2 rt phong ph, sn c ty thuc tng a phng, sn xut nhin liu ny vn cha thc s c hiu qu kinh t do cc ro cn v mt k thut ch bin. Hai loi nhin liu in hnh ca NLSH th h 2 l cellulosic ethanol and Fischer Tropsch fuels u cha c sn xut i tr. Tuy nhin, c th thy r NLSH th h 2 gim r rt pht thi kh CO2, khng cnh tranh vi cy lng thc v c th cnh tranh c vi xng v du c ngun gc t du m trong tng lai nu c cc t ph trong cng ngh. Vic dng to sn xut nhin liu sinh hc thay th du m ging nh mt mi tn bn trng hai ch: va to ra nng lng va lm sch mi trng. Mi t bo to l mt nh my sinh hc nh, s dng qu trnh quang hp chuyn ha CO2 v nh sng mt tri thnh nng lng d tr trong t bo v to ra cc sn phm th cp c gi tr cao. Hot ng chuyn i ca chng hiu qu n mc sinh khi ca chng c th tng gp nhiu ln trong mt ngy. Ngoi ra, trong qu trnh quang hp, to cn sn xut ra du ngay trong t bo ca chng. Trn cng mt n v din tch, lng du m to to ra nhiu gp 30 ln lng du t u nnh. ng thi to c th tng kh nng sn xut du bng cch b sung kh CO2 trong qu trnh nui trng chng hoc s dng cc mi trng giu cht hu c (nh nc thi) nui trng (Demirbas, 2009b, Demirbas, 2010). iu ny va to ra NLSH, va lm gim lng CO2 cng nh lm sch mi trng. Hin nay, NLSH dng ch yu trong vn ti l ethanol sinh hc (thay th 1 phn xng), diesel sinh hc (thay th 1 phn diesel du m) v NLSH c pht trin t toth h NLSH th ba ang c quan tm nghin cu.
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cng lun vn tt nghip

1.2 Tng quan v to. Vi to (Microalgae) l tt c cc to (Algae) c kch thc hin vi. Mun quan st chng phi s dng ti knh hin vi.Trong s khong 50 000 loi to trn th gii th vi to chim n khong 2/3. Vi to ch yu thuc v cc chi trong cc ngnh sau y: -Ngnh To lc (Chlorophyta): Cc chi Closterium, Coelastrum, Dyctyosphaerium, Scenedesmus, Pediastrum, Staurastrum, Dunaliella, Chlamydomonas, Haematococcus, Tetraselmis, Chlorella,... - Ngnh To lng roi lch (Heterokontophyta) : Cc chi Melosira, Asterionella, Cymatopleurra, Somphonema, Fragilaria, Stephanodiscus, Navicula, Malomonas, Dinobryon, Peridinium, Isochrysis, Chaetoceros, Phaeodactylum, Skeletonema, Nitzschia...... - Ngnh To mt (Euglenophyta): Cc chi Phacus, Trachelomonas, Ceratium... - Ngnh To (Rhodophyta): Cc chi Porphyridium, Rhodella... 1.2.1 Vai tr ca vi to trong t nhin v trong i sng nhn loi: To ni chung v vi to ni ring c vai tr rt quang trng trong t nhin v trong i sng nhn loi. Chng ta bit rng i dng chim 71% din tch b mt Tri t. Mt s tc gi Hoa K cho rng hng nmg to c th tng hp ra trong i dng 70280 t tn cht hu c. Trong cc thy vc nc ngt to cung cp xy v hu ht thc n s cp cho c v cc ng vt thy sinh khc. To gp phn bo v mi trng nui thy sn bng cch tiu th bt lng mui khong d tha. Canh tc bin l nhm trng v thu hoch cc to sinh khi ln v c hm lng dinh dng cao. Nhiu to bin cn khai thc sn xut thch (agar), alginate, sn phm giu iod... Nhiu to n bo c nui trng cng nghip to ra nhng ngun thc n cho ngnh nui tm hay thuc b tr giu protein , vitamin v vi khong dng cho ngi. Mt s vi to c dng sn xut carotenoid, astaxanthin, cc acid bo khng bo ha... To silic to ra cc m diatomid, l loi nguyn liu xp, nh, mn c dng trong nhiu ngnh cng nghip.

cng lun vn tt nghip

To phn b ht sc rng ri khp mi ni, t nh ni cao n y bin su. Nhng to sng lp nc pha trn c gi l To ph du (Phytoplankton) cn nhng to sng bm di y thy vc, bm trn cc vt sng hay thnh tu thuyn c gi l To y (Phytobentos). Dng to cng sinh vi nm thnh a y cng l dng phn b rt rng ri v nhiu loi c khai thc dng lm dc phm, nc hoa, phm nhum v cc mc ch kinh t khc (hin bit ti 20 000 loi a y thuc 400 chi khc nhau). 1.2.2 Vai tr ca vi to trong sn xut nhin liu sinh hc. To c cu trc t bo n gin, tc sinh sn cc k nhanh v hm lng cht bo li rt ln nn c u tin la chn lm nguyn liu sn xut nng lng thay th biodiesel.
_u v nhc im ca sn xut NLSH t sinh khi vi to

u im

Nhc im

Tc sinh trng cao Khng i hi cc ngun nc ngt cho nui trng (c th trng trong nc thi, nc l, nc bin). Nhu cu dng nc t hn cc loi cy lng thc. Hiu sut cao trong chuyn ha kh thi CO2 thnh sinh khi, gip gim pht thi kh nh knh.

Mt sinh khi ca vi to thp, v s hn ch ca nh sng thm nhp su vo trong lp nc. Vn u t ban u cao hn cc loi cy trng khc

cng lun vn tt nghip

Hiu sut cho du cao hn so vi t cy lng thc nn din tch cho nui trng c th thu hp hn so vi cc cy khc v c th pht trin cc vng t cn ci, hoang ha, nhng c nh sng v nc.

Sn xut diesel sinh hc t sinh khi vi to l hng i m nhiu nc trn th gii ang hng n. u th vt tri ca cng ngh ny l khng cnh tranh t canh tc nng nghip. To chng t l ngun nguyn liu bn vng cho sn xut NLSH. So vi NLSH th h 2, to-nguyn liu cho NLSH th h th 3, cho nng sut sinh khi v cho du cao hn t 30-100 ln (Demirbas, 2009a, Chisti, 2008b). 2.TNG QUAN TI LIU. 2.1Trn th gii. _1978, USA, Phng th nghim nng lng ti to quc gia chng trnh cc loi thy sn nghin cu hn 3000 loi to tm kim cc loi to c lng du cao, nng sut cao v s lng ln thay th cho nhin liu ha thch. _Chan Yoo v cng s qua nghin cu ca mnh cho ra kt lun sau: Trong ba loi c kho st th Scenedesmus sp l loi t nng sut sinh khi cao nht. Sinh khi ti a t c l 3.13g/L vo ngy th 14, nng sut sinh khi t 217.50 11.24 mg sinh khi kh/L.ngy Hm lng lipid tng trong vi to t t 6.25% ti 25.79% khi lng kh. _Morais v Costa cho kt qu: Nng sut ti a ca hai loi Chlorella sp. v Scenedesmus sp. vi 6% CO2 ln lt l 87 v 85 mg sinh khi kh/L.ngy. Hn na chng c th sng trong mi trng cha CO218%. Nng sut sinh khi ca Scenedesmus obliquus l 140mg sinh khi kh/L.ngy vi nng 12% CO2.
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_Lee v cc cng s xut rng khi tng mt t bo ban u th s tng c sc chu ng ca vi to i vi nng CO2 cao v gim c thi gian thch nghi. Nh vy, nu B. braunii c cho thch nghi vi nng CO2 thp trc , th mc sinh trng v sn lng sinh khi s c th gia tng m khng cn tri qua thi k thch nghi. _Guillard xc nh nng nitrogen trung bnh trong mi trng nui cy Chlorella vulgaris l 1.50g/L. Attilio v cng s nghin cu nng 0.750 v 0.375g/L so snh, cho kt lun hm lng nitrogen thp trong mi trng nui cy lm tng ng k s tch ly lipidtrong vi to. 2.2Ti Vit Nam. _Nm 2008, TS. Trng Vnh v cc cng s H Nng Lm TP.HCM va c nhng kt qu nghin cu cho thy to bin Chlorella c nhiu trin vng ng dng ti VN, l ngun sn xut biodiesel phong ph m khng xm hi an ninh lng thc nh nhng loi cy trng ly du biodiesel khc. Nghin cu s dng ngun to ging Chlorella trong nc, c cung cp t Khoa Thu sn Trng H Nng Lm TP.HCM, Khoa Thu sn Trng H Cn Th v Trung tm Quc gia ging Hi sn Nam B. Th nghim cho thy to Chlorella cho du c mu vng sm, nng sut chuyn i du thnh biodiesel l 97% sau 2 gi phn ng. TS. Trng Vnh ngh, nn nhp cc ging to hm lng du cao cc n v thu sn nghin cu trin khai nui trng cc vng ngp mn, hoang ho. ng thi cn nghin cu ng dng cc thit b nui quang hp, chit tch du t ch to, gim gi thnh sn xut biodiesl trong tng lai. 3. PHNG PHP VT LIU. 3.1Phng php. Qui trnh p dng trong ti ny nh sau:
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-Nui 8 loi to nc ngt trn mi trng BB v MWH. -Xc nh lng du c trong to bng Nile red - UV hunh quang, chn 3 con. -Tip tc cy v tng sinh 3 to chn trong bnh 20l. -Thc hin cc th nghim to stress kho st s tch ly lipid:s thay i nhit thp, cng chiu sng mnh, gim lng Nit, tng lng mui, tng lng st cho kt qu tng lipid ln 30%- 40% khi lng kh. Phng php chit du t to: -Soxhlet v phng php un hon lu, my c quay thu hi dung mi. - Thu sinh khi to v tch du t sinh khi theo kiu truyn thng l ly tm, tch phn ni v phn kt lng, lc mng hoc dng sng siu m. -Phng php s dng hexane lm dung mi ha tan. -Phng php chit lng ti hn Supercritical fluid extraction. -Ly trch bng enzymephas hy t bo vi nc l dung mi ha tan. -Osmotic shock l gy sc bng s kh t ngt trong p sut thm thu lm v t bo, phng thch thnh phn t bo nh du. 3.2 Vt liu 3.2.1 Ha cht _Mi trng BB: Bold- Basal NaNO3 MgSO4.7H2O NaCl K2HPO4 KH2PO4 CaCl2.2H2O dd vi lng nc ct 0.25g 0.075g 0.025 0.075g 0.175g 0.025g 1.0ml 1000ml

+Thnh phn vi lng

cng lun vn tt nghip

EDTA ZnSO4.7H2O MnCl2.4H2O MoO3 CuSO4.5H2O Ca(NO3)2.6H2O FeSO4.7H2O H3BO3 nc ct

50g 8.82g 1.44g 0.71g 1.57g 0.49g 4.98g 11.42g 1000ml

_Mi trng MWH: Modified Woods Hole CaCl2.2H2O MgSO4.7H2O NaHCO3 K2HPO4.3H2O NaNO3 Na2SiO3.5H2O dd vi lng nc ct 0.0368g 0.037g 0.0126g 0.0114g 0.085g 0.0212g 1.0ml 1000ml

+Thnh phn vi lng EDTA FeCl3.6H2O CuSO4.5H2O ZnSO4.7H2O CoCl2.6H2O MnCl2.4H2O Na2MoO4.2H2O H3BO3 nc ct 3.2.2 Thit b -Knh hin vi -Soxhlet -My c quay
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4.36g 3.15g 0.01g 0.022g 0.01g 0.18g 0.006g 1g 1000ml

cng lun vn tt nghip

-Chai nc bin -Bnh cy 15-20l -a petri, que cy, n cn. IV. D KIN KT QU T C. Theo cng nu trn, d kin kt qu thu c nh sau: -Xc nh c t nht l 3 ging to c tim nng tng sinh v tch ly lipid cao. -Xc nh ng cong tng trng, tc tng trng v tng lng lipid thu c. -Kho st c iu kin ti u no thch hp cho nui cy tng sinh v tch ly lipid ca to.

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