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产肠杆菌内切葡聚糖酶基因工程酿酒酵母生产纤维素乙醇。

Cellulosic ethanol production by Zymomonas mobilis harboring an endoglucanase gene from Enterobacter cloacae.

机构信息

Department of Biotechnology, Anna University Tiruchirappalli, Tiruchirappalli 620024, Tamil Nadu, India.

出版信息

Bioresour Technol. 2011 Feb;102(3):2585-9. doi: 10.1016/j.biortech.2010.09.110. Epub 2010 Oct 23.

Abstract

Enterobactercloacae was isolated from the gut of the wood feeding termite, Heterotermes indicola, and a 2.25-kb fragment conferring cellulase activity was cloned in Escherichia coli. The cloned fragment contained a 1083-bp ORF which could encode a protein belonging to glycosyl hydrolase family 8. The cellulase gene was introduced into Zymomonas mobilis strain Microbial Type Culture Collection centre (MTCC) on a plasmid and 0.134 filter paper activity unit (FPU)/ml units of cellulase activity was observed with the recombinant bacterium. Using carboxymethyl cellulose and 4% NaOH pretreated bagasse as substrates, the recombinant strain produced 5.5% and 4% (V/V) ethanol respectively, which was threefold higher than the amount obtained with the original E.cloacae isolate. The recombinant Z. mobilis strain could be improved further by simultaneous expression of cellulase cocktails before utilizing it for industrial level ethanol production.

摘要

从木质素取食的土白蚁 Heterotermes indicola 的肠道中分离到阴沟肠杆菌,并在大肠杆菌中克隆了一个可赋予纤维素酶活性的 2.25kb 片段。该克隆片段包含一个编码属于糖苷水解酶家族 8 的蛋白质的 1083bpORF。纤维素酶基因被引入到质粒上的运动发酵单胞菌微生物类型培养中心(MTCC)菌株中,并观察到重组菌的纤维素酶活性为 0.134 滤纸活性单位(FPU)/ml。使用羧甲基纤维素和 4%NaOH 预处理的蔗渣作为底物,重组菌分别产生 5.5%和 4%(V/V)的乙醇,比原始阴沟肠杆菌分离株的产量高三倍。在将重组运动发酵单胞菌菌株用于工业水平的乙醇生产之前,通过同时表达纤维素酶混合物,可以进一步改进该重组菌。

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