Kanayama H, Sode K, Karube I
Tokyo Research Laboratory, Japan Synthetic Rubber Co., Ltd., Asao-ku Kawasaki.
Appl Biochem Biotechnol. 1987 Aug;15(2):97-106. doi: 10.1007/BF02801311.
Citrobacter freundii genes that complemented Escherichia coli hyd-(hydrogenase activity) mutation were cloned in plasmids pCBH4 (6.2 kb) and pCBH6(5.7 kb). Hydrogen evolution by the transformant E. coli HK-8(pCBH4 or pCBH6) was investigated. The optimum culture temperature of recombinant E. coli cells for hydrogen evolution from glucose was in the neighborhood of 18 degrees C. The recombinant E. coli cells cultured at this condition showed a several-fold increase of hydrogen evolution, as compared with that of the wild-type cells. The plasmid-retention stability of this recombinant E. coli was extremely high, especially plasmid pCBH4, which was completely retained during 2 wk without any restriction. Hydrogen production by immobilized recombinant E. coli was then investigated using cells cultured at 18 degrees C. The hydrogen evolution rate from glucose and Lennox-broth were about twofold higher than that of E. coli C600, and this high hydrogen evolution rate was maintained for more than 1 mo.
弗氏柠檬酸杆菌中可互补大肠杆菌hyd-（氢化酶活性）突变的基因被克隆到质粒pCBH4（6.2 kb）和pCBH6（5.7 kb）中。对转化体大肠杆菌HK-8（pCBH4或pCBH6）的析氢情况进行了研究。重组大肠杆菌细胞从葡萄糖析氢的最适培养温度在18℃左右。在此条件下培养的重组大肠杆菌细胞与野生型细胞相比，析氢量增加了几倍。这种重组大肠杆菌的质粒保留稳定性极高，尤其是质粒pCBH4，在2周内完全保留，没有任何限制。然后使用在18℃培养的细胞研究固定化重组大肠杆菌的产氢情况。从葡萄糖和Lennox肉汤中的析氢速率比大肠杆菌C600高约两倍，并且这种高析氢速率维持了1个月以上。
