殏 檪檪 殏 檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪 China Biotechnology 202 32 3 9-99 殏 殏 8 Mitchell Bieniek Mitchell Bieniek 2 Garrett Flack 2 * 00086 2 60506 Clostridium acetobutylicum AS. 7 Clostridium acetobutylicum AS. 32 Clostridium acetobutylicum AS. 34 Clostridium beijerinckii NCMIB 8052 NCMIB8052 Q85 20-2-29 202-02-24 * lvjihua@ rdfz. cn 5%
92 China Biotechnology Vol. 32 No. 3 202 2. 3. 4 0g 3 00ml 5ml 0. 5mol /L 24 % 24 ph NaOH Ca OH 2 6. 5. 4 4. 4. 5-6 45ml. 4. 2 35 80 0h 24h 48h 72h OD 600nm. 4. 3 0h 24h 48h 72h min 5 000r /min. Clostridium acetobutylicum AS. 7 Clostridium acetobutylicum AS. 32 Clostridium acetobutylicum AS. 34 Clostridium beijerinckii NCMIB 8052. 2 SoftMax SPECTRA MAX 90 5424AJ770005 Eppendorf 200 series Agilent Technologies HDL APPARATUS ZHWY-202 2. 3. 3. g /L AS. 32 NCIMB 8052 40 5 NH 4 2 SO 4 2 KH 2 PO 4 0. 75 K 2 HPO 4 a d 3H 2 O 0. 75 NaCl MgSO 4 7H 2 O 0. 4 MnSO 4 H 2 O 24 OD 0. 0 FeSO 4 7H 2 O 0. 0 2 48. 3. 2 g /L 40 5 NH 4 2 SO 4 2 KH 2 PO 4 0. 75 K 2 HPO 4 3H 2 O 0. 75 NaCl MgSO 4 7H 2 O 0. 4 MnSO 4 H 2 O 0. 0 FeSO 4 7H 2 O 0. 0 2 ph. 3. 3 g /L 7 30 0 5 NH 4 2 SO 4 2 KH 2 PO 4 0. 75 K 2 HPO 4 3H 2 O 0. 75 NaCl MgSO 4 7H 2 O 0. 4 HPX-87H 5 0. 05 mmol /L H 2 SO 4 0. 5 ml /L 2 2. 2.. OD 2a 2d MnSO 4 H 2 O 0. 0 FeSO 4 7H 2 O 0. 0 2 ABE 8
202 32 3 93 AS. 34 24 OD 2. 3. 2 0. 9g /L AS. 7 48 AS. 32 NCIMB8052 2.. 2 Fig. The change of OD and butanol concentration verses time 2.. 2 8 AS. 7 AS. 32 NCMIB 8052 AS. 7 AS. 7 AS. 32 24h ph NCMIB8052 48h AS. 7 2.. 48h AS. 7 2 0. 3g /L AS. 7 2c AS. 32 NCMIB 8052 ABE AS. 7
94 China Biotechnology Vol. 32 No. 3 202 Fig2 2 The change of other substances concentration verses time 2. 2 NCMIB8052 3a AS. 32 AS. 32 AS. 7 NCMIB8052 48h OD 48h 2. 2. 3 OD 2.. 0. 9g / L 4 AS. 34. 7 2. 2. 2 AS. 34 3b 2. 2. 2 S AS. 34 AS. 32 NCMIB8052 24 AS. 34 OD NCMIB8052 OD 3 2. 3. 2 AS. 34 3c 3d AS. 7
202 32 3 95 Fig3 3 The change of OD and xylose concentration verses time AS. 34 g /L a c d AS. 32 AS. 7 NCMIB 0 ~ 72 2. 3. 5 24 5 AS. 34 AS. 7 AS. 32 NCMIB8052 4a 4d AS. 32 NCMIB8052 NCMIB8052 0. 5g /L 2.. 2 AS. 7 NCMIB8052 AS. 32 NCMIB8052 NCMIB8052 AS. 34 NCMIB8052 2. 3
96 China Biotechnology Vol. 32 No. 3 202 Fig4 4 The change of other substances concentration verses time Fig5 5 The change of sugar concentration and xylose concentration verses time 2. 3. 2 6 NCMIB8052 AS. 32 AS. 34 5b 6 6b 24
202 32 3 97 2b 6b AS. 34 AS. 34 3. 5g /L AS. 34 AS. 7 AS. 32 NCMIB8052 AS. 7 AS. 34 2 AS. 7 2. 2 24 24h AS. 7 OD 5a AS. 34 2. 5g /L AS. 34 24 OD AS. 34 Fig6 6 The change of other substances concentration verses time 2. 4 IMSA AS. 32 AS. 34 NCMIB8052 ph NaOH Ca OH 2 7
98 China Biotechnology Vol. 32 No. 3 202 8 Ca OH 2 NCMIB8052 ABE 8 NCMIB8052 7 NCMIB8052 2 Fig. 7 AS. 32 8g /L NCMIB8052 AS. 32 AS. 34 24 8 8a NaOH 7 8b Ca OH 2 AS. 34 ABE Fig8 8 The fermentation result with hydrolysate as substrate 3 NCMIB8052 Max McGee.. 2009 38 7 059-063. Wu X Gu L L Shen L Z et al. Study on the developing trend of fuel ethanol and ethanol gasoline for motor vehicles. Applied Chemical Industry 2009 38 7 059-063. 2.. 2003 7 2 9-2. Mei Y F. Gansu Chemical Industry 2003 7 2 9-2.
202 32 3 99 3 Johnson J L Toth J Santiwatanakul S et al. Cultures of Clostridium acetobutylicum from Various Collections Comprise Clostridium acetobutylicum Clostridium beijerinckii and two other distinct types based on DNA-DNA reassociation. International Journal of Systematic Bacteriology 997 47 2 420-424. 4 Gu Y Li J Zhang L et al. Improvement of xylose utilization in Clostridium acetobutylicum via expression of the tala gene encoding transaldolase from Escherichia coli. Journal of Biotechnology 2009 43 4 284-287. 5. Clostridium beijerinckii NCIMB 8052 8052 strains with high concentration butanol tolerance in cassava fermentation. Liquor-Making Science &Technology 200 9. 200 9 8 Thaddeus Chukwuemeka Ezeji Nasib Qureshi Hans Peter 36-39. Su H F Yang D F. Screening of Clostridium beijerinckii NCIMB Blaschek. Bioproduction of butanol from biomass from genes to bioreactors. Current Opinion in Biotechnology 2007 8 220-227. 36-39. 6 Isar J Rangaswamy V. Improved n-butanol production by solvent tolerant Clostridium beijerinckii. Biomass and Bioenergy 202 37 9-5. 7 Maddox I S Steiner E Hirsch S et al. The Cause of Acid Crash and Acidogenic Fermentations During the Batch Acetone-Butanol-Ethanol ABE Fermentation Process. J Mol Microbiol Biotechnol 2000 2 95-00. Study on the Ability of Butanol Production of Different Bacteria with the Fermentable Sugar GUO Yong-an TENG Ya-qun ZHU Ouhaodi DAU Yi-chen ZHA Jing-jing ZHU Xu ZENG Xiao XING Xiao-xue Mitchell Bieniek 2 Garrett Flack 2 LV Ji-hua The High School Affiliated to Renmin University of China Beijing 00080 China 2 Illinois Mathematics and Science Academy Illinois 60506 U. S. A Abstract With the development of the new generation of biomass energy it has become a popular topic that to use fermentable sugar and Clostridia to produce butanol. The study is going to use Clostridium acetobutylicum AS. 7 Clostridium acetobutylicum AS. 32 Clostridium acetobutylicum AS. 34 and Clostridium beijerinckii NCMIB 8052 which can produce butanol in various sources of sugar by fermentation. By comparing growth behaviors ratio of using sugars yield of butanol and by-product and endurance of butanol and xylose we are going to find out the most suitable Clostridium which can be used to the industrial. In the experiment NCMIB8052 is the most outstanding Clostridium as it has the highest yield relative high endurance and the ability using multiple sugars. Key words Biomass Butanol Fermentation Clostridium Endurance Xylose