产氢菌株Clostridium sp.T7的快速筛选

A Rapid Screening for Isolation of Hydrogen-Producing Bacterium Clostridium sp.T7

取自潮间带的污泥分别在不同温度下(80、100、121,℃)进行热休克预处理,富集产氢菌群并测定其产氢量,利用变性梯度凝胶电泳(DGGE)分析混合菌群组成.结果表明:3种热处理条件下混合菌群的产氢量都要高于对照未处理菌群.DGGE图谱表明,与80、100,℃热休克处理混合菌群相比,经121,℃热休克处理富集的混合菌群,其电泳条带最少,测序结果发现该混合菌群中包括产氢菌Clostridium sp..从该混合菌群中纯化并鉴定了1株产氢菌株Clostridiumsp.T7(登录号HM104461).培养温度对菌株T7产氢有一定影响,温度在25～55,℃范围内菌株Clostridium sp.T7都能产氢,最适产氢温度是35,℃.

To determine the effects of heat-shock pretreatment on hydrogen production and hydrogen-producing microbial community,sludge from the intertidal zone was pretreated with three different heat treatment temperature,respectively 80,℃,100,℃ and 121,℃.The results showed that hydrogen production of mixed culture by three heat-shock pretreatments was higher than those in the control group.The result of denaturing gradient gel electrophoresis（DGGE）showed that electro-phoretic bands of mixed culture pretreated by heat-shock temperature of 121,℃ were the least,compared with those by heat-shock pretreatment of 80,℃ and 100,℃.Although the composition of microbial community isolation by heat-shock tempera-ture of 121,℃ was simple,this heat treatment temperature was favorable to enrich the dominant hydrogen-producing bacterium,i.e.Clostridium sp..A hydrogen-producing bacterium strain T7（accession number HM104461）was isolated from the mixed culture by heat-shock pretreatment of 121,℃.The effect of culture temperature on hydrogen production of Clostrid-ium sp.T7 was determined.The strain was able to produce hydrogen over a wide range of culture temperature from 25,℃ to 55,℃,with an optimum culture temperature of 35,℃.