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1 微生物学通报 DEC 20, 2009, 36(12): 1871~1875 Microbiology 2009 by Institute of Microbiology, CAS 简报正交实验优选光合细菌混合菌群产氢的最佳条件 * 崔宝臣张国欣侯博邵姝媛 ( ) 摘要 : 对光合细菌混合菌群产氢影响因子进行了实验研究通过单因素实验和正交实验, 系统考察了碳源氮源碳源浓度氮源浓度初始 ph 值光照方式接种量等因素对产氢量的影响, 实验得出最佳工艺条件为 : 采用 3 号菌群, 碳源为葡萄糖, 碳源浓度为 3 g/l, 氮源为尿素, 氮源浓度为 9 g/l, 接种量为 10%, ph 值为 8.5, 光照方式为 12 h 光照 -12 h 黑暗交替光照, 培养温度为 30 C 菌种碳源碳源浓度氮源是影响产氢量的重要因素关键词 : 光合细菌, 混合菌群, 产氢, 正交实验 Optimization of Hydrogen Production Conditions of Photosynthetic Bacteria Group by Orthogonal Test CUI Bao-Chen * ZHANG Guo-Xin HOU Bo SHAO Shu-Yuan (College of Chemistry and Chemical Engineering, Daqing Petroleum Institute, Daqing, Heilongjiang , China) Abstract: The influence factors for hydrogen production using photosynthetic bacteria group were studied. The effects of carbon, nitrogen, carbon concentration, nitrogen concentration, initial ph value, illumination mode and inoculation amount on hydrogen production were investigated by experiment of single factor and orthogonal experiment. The optimum process conditions were as follows: the best bacteria group was number 3, the glucose was carbon source and concentration of carbon source was 3 g/l; the urea was nitrogen source and concentration of nitrogen source was 9 g/l; the inoculation amount was 10%; initial ph value was 8.5; the illumination mode was 12 h light and 12 h dark alternation; incubation temperature was 30 C. The main factors affecting hydrogen production were strain, carbon, carbon source concentration and nitrogen suorces. Keywords: Photosynthetic bacteria, Mixed bacteria group, Hydrogen production, Orthogonal test,,,, [1], (Rhodobacter sphaeroides) [2] (Rhodopseudomonas)H, [3] C/N Barbosa [4] 基金项目 :(No. GZ08B205) * 通讯作者 :Tel: ; : [email protected] 收稿日期 : ; 接受日期 :

2 1872 微生物学通报 2009, Vol.36, No.12 3, ph,, 1 材料与方法 1.1 菌种与培养基 123 : NH 4 Cl 0.1 g, NaHCO g, K 2 HPO g, CH 3 COONa 0.3 g, MgSO 4 7H 2 O 0.02 g, NaCl 0.15 g, 1 ml, 1 ml, 97 ml : (C 5 H 8 NO 4 Na) 0.1 g, NaHCO g, K 2 HPO g, CH 3 COONa 0.3 g, MgSO 4 7H 2 O 0.02 g, NaCl 0.15 g, 1 ml, 1 ml, 97 ml 1.2 试剂与仪器 ; SP3420 ; LRH-250 ; HWS26 ; PHS-25 ph 1.3 实验方法 10%, 50 C 30 min,,, 100 W, 30 C0.5 C 8 d,, = 1000/, : ml H 2 /(L-); =/( ), : ml H 2 /(L-h) 1 2, ph, 3 Wakayama [5], 4, 5 10%,, 1 Table 1 Effect of different carbon and nitrogen on hydrogen production Nutrient Carbon Nitrogen Category H 2 Hydrogen amount [mlh 2 /(L-culture)] The highest hydrogen rate [mlh 2 /(L-culture h)] 结果与讨论 2.1 影响产氢量的主要参数的确定, ph 1 图 1 葡萄糖浓度对产氢影响 Fig. 1 Effect of glucose concentration on hydrogen production

3 : 1873 图 2 谷氨酸钠浓度对产氢影响 Fig. 2 Effect of nitrogen concentration on hydrogen production 图 3 不同初始 ph 值对产氢影响 Fig. 3 Effect of initial ph value on hydrogen production 图 5 接种量对产氢影响 Fig. 5 Effect of inoculation amount on hydrogen production 2.2 参数优化正交实验结果, ph 9, 3, 2 L 27 (3 13 ) 9 3 3,,, ph 4, P 0.05,,, 3 结论图 4 光照方式对产氢影响 Fig. 4 Effect of illumination mode on hydrogen production : 12/12: 12 h, 12 h ; 16/8: 16 h, 8 h ; 24 h:. Note: 12/12: 12 h light, 12 h dark alternation; 16/8: 16 h light, 8 h dark alternation; 24h: Continuous illumination. 1), ph, 2) 9, : 3 3 g/l 9 g/l 10%pH

4 1874 微生物学通报 2009, Vol.36, No.12 Levels A Strains B Carbon C Nitrogen Table 2 表 2 正交实验水平因素 Levels and factors of orthogonal experiment D Carbon concentration (g/l) Factors E Nitrogen concentration (g/l) F Inoculation amount (%) G ph Initial ph value H Illumination mode I Incubation temperature ( C) * * * 35 : 1 * : 24 h ; 2 * : 16 h, 8 h ; 3 * : 12 h, 12 h. Note: 1 * : 24 h continuous illumination; 2 * : 16 h light, 8 h dark alternation; 3 * :12 h light,12 h dark alternation. No. 表 3 L 27 (3 13 ) 正交实验表与结果 Table 3 L 27 (3 13 ) orthogonal experiment table and results Factors A B C D E F G H L Hydrogen amount [mlh 2 /(L-culture)] R

5 : 1875 表 4 方差分析 Table 4 Variance analysis Source SS DF MS F P A B C D E F G H I Error h -12 h 30 C, 参考文献 [1],,,.., 2004, 55 (): [2],,,. H., 1994, 21(5): [3],,,.., 2007, 28( 2): [4] Barbosa MJ, Rocha JMS, Tramper J, et al. Acetate as a carbon source for hydrogen production by photosynthetic bacteria. Journal of Biotechnology, 2001, 85(1): [5] Wakayama T, Nakada E, Asada Y, et al. Effect of light/dark cycle on bacterial hydrogen production by Rhodobacter sphaeroides RV. Applied Biochemistry and Biotechnology, 2000, 1-9(84/86): 年中科院微生物所期刊联合编辑部联合征订全面启动! 征订启事 ( 4 ), 55.00, 660 ISSN ; CODEN WSHPA82-504; BM67 ( 25 ), 65.00, 780 ISSN ; CODEN SGXUED82-13; BM5608 ( 20 ), 48.00, 576 ISSN ; CODEN WSWPDI2-817; BM413 ( 15 ), 80, 480 ISSN ; CODEN JXUUAE2-499; Q723,. (100101) 1 3 B401 ; (010) ; [email protected]

H 2 SO ml ml 1. 0 ml C 4. 0 ml - 30 min 490 nm 0 ~ 100 μg /ml Zhao = VρN 100% 1 m V ml ρ g

H 2 SO ml ml 1. 0 ml C 4. 0 ml - 30 min 490 nm 0 ~ 100 μg /ml Zhao = VρN 100% 1 m V ml ρ g 16 6 2018 11 Chinese Journal of Bioprocess Engineering Vol. 16 No. 6 Nov. 2018 doi 10. 3969 /j. issn. 1672-3678. 2018. 06. 004 1 2 1 1 1. 330200 2. 330006 4 Box-Behnken 1 25 g /ml 121 92 min 2 6. 284%