厌氧发酵产氢细菌的厌氧操作与紫外线诱变技术

Anaerobic operation and UV-radiation mutagenesis for obligate anaerobic fermentative hydrogen-producing bacteria

结合几种厌氧操作,即Hungate滚管分离法、厌氧培养瓶分离法、平板夹层厌氧法和培养皿平板分离法,对乙醇型产氢发酵细菌进行紫外诱变及突变体分离筛选,比较证明在提供厌氧操作条件下(厌氧操作箱),直接涂布平板分离法是培养、分离、筛选严格厌氧产氢突变菌株的最佳方法,通过对分离培养基成分的调整及观测菌落的生长突变及分离效果,确定紫外分离厌氧产氢突变菌株的分离培养基。同时确定乙醇型发酵产氢细菌紫外辐射的最适照射时间控制在3 m in之内,突变菌株遗传稳定性的传代次数至少控制在6次。紫外诱变获得的高效稳定产氢突变体的产氢能力比对照菌株提高40%—65%。

Ethanol-type hydrogen-producing bacteria were mutated by UV-radiation and screened mutants in combined with several anaerobic operations, such as Hungate technique, anaerobic culture flask, plate interlay anaerobic isolation and indirect petri-dish plate isolation. The results indicate that the optimal method is direct spread petri-dish plate isolation in cultivating, isolating and screening obligate anaerobic mutants under anaerobic conditions （anaerobic operation box）. The isolation media was confined by regulating the composition of isolated media and observing the effects of isolation and mutated amount of hydrogen-producing mutants colonies. Moreover, the experiment proves that the optimal UV-radiating time and pass generation times are controlled within 3 min and at least 6 times for ethanol-type fermentative H2-production bacteria, respectively. The H2-producing ability of mutants screened by UV is improved by 40%-65% than that of control strains.