Klebsiella sp.WL1316乙醇脱氢酶基因过表达提高乙醇生产效率  

Overexpression of alcohol dehydrogenase in Klebsiella sp.WL1316 to improve hydrogen production efficiency

作　　者：周柱[1] 查凡 张琴[1] 陈振[1] 陈翔宇 胡永安 ZHOU Zhu;ZHA Fan;ZHANG Qin;CHEN Zhen;CHEN Xiang-yu;HU Yong-an(College of Biological and Food Engineering,Anhui Polytechnic University,Wuhu 241000,Anhui Province,China)

机构地区：[1]安徽工程大学生物与食品工程学院,安徽芜湖241000

出　　处：《化学工程》2022年第10期1-7,共7页Chemical Engineering(China)

基　　金：安徽省重点研究与开发项目(201904a07020003);安徽省自然科学基金项目(2108085MB34);安徽省大学生创新项目(S202010363252)。

摘　　要：为获得能用于纤维素乙醇高效生产的基因重组菌株,文中针对能利用木质纤维素水解液的Klebsiella sp.WL1316,对其乙醇脱氢酶adh基因进行了克隆和同源过表达。成功构建adh-pET-28a-Klebsiella sp.WL1316重组菌株,通过对菌株发酵稻草水解液生产乙醇过程的监测确定该重组菌适宜的发酵时间为48 h。结果表明:进一步通过单因素和响应面试验获得菌株乙醇发酵生产的优化工艺参数为IPTG诱导浓度0.89 mmol/L,发酵起始pH值7.59,起始还原糖质量浓度60.60 g/L。在此优化工艺条件下进行验证实验,乙醇产量达(8.28±0.55)g/L,与模型的理论预测值较接近,说明建立的模型是切实可行的,并且,该乙醇产量为同等发酵条件下野生菌的2.26倍,说明adh基因的过表达促进了乙醇产量的显著提高。To obtain a genetically recombinant strain that can be used for efficient cellulosic ethanol production,the alcohol dehydrogenase adh gene of Klebsiella sp.WL1316 with lignocellulose hydrolysate-utilizing ability,was cloned and overexpressed.The recombinant strain adh-pET-28a-Klebsiella sp.WL1316 was successfully constructed,and the suitable fermentation time of the recombinant strain was determined to be 48 h by monitoring the process of ethanol production from rice straw hydrolysate fermentation.The results show that the optimized technological parameters of fermentative ethanol production were obtained via single factor and response surface tests as follows:the IPTG induced concentration is 0.89 mmol/L,the initial pH value of fermentation is 7.59,and the initial reducing sugar concentration is 60.60 g/L.The verification experiment was carried out under the optimized parameters,and the ethanol yield is reached(8.28±0.55)g/L,which is close to the theoretical predicted value of the model,indicating the established model is feasible,moreover,such ethanol yield is 2.26 times that of wild strain under the same fermentation condition,indicating the overexpression of adh gene improves the ethanol production significantly.

关 键 词：Klebsiella sp. 乙醇脱氢酶基因 过表达 乙醇 发酵 

分 类 号：Q939.9[生物学—微生物学]