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通过代谢途径工程开发一种能够发酵阿拉伯糖的运动发酵单胞菌菌株。

Development of an arabinose-fermenting Zymomonas mobilis strain by metabolic pathway engineering.

作者信息

Deanda K, Zhang M, Eddy C, Picataggio S

机构信息

National Renewable Energy Laboratory, Golden, Colorado 80401, USA.

出版信息

Appl Environ Microbiol. 1996 Dec;62(12):4465-70. doi: 10.1128/aem.62.12.4465-4470.1996.

Abstract

The substrate fermentation range of the ethanologenic bacterium Zymomonas mobilis was expanded to include the pentose sugar, L-arabinose, which is commonly found in agricultural residues and other lignocellulosic biomass. Five genes, encoding L-arabinose isomerase (araA), L-ribulokinase (araB), L-ribulose-5-phosphate-4-epimerase (araD), transaldolase (talB), and transketolase (tktA), were isolated from Escherichia coli and introduced into Z. mobilis under the control of constitutive promoters that permitted their expression even in the presence of glucose. The engineered strain grew on and produced ethanol from L-arabinose as a sole C source at 98% of the maximum theoretical ethanol yield, based on the amount of consumed sugar. This indicates that arabinose was metabolized almost exclusively to ethanol as the sole fermentation product, with little by-product formation. Although no diauxic growth pattern was evident, the microorganism preferentially utilized glucose before arabinose, apparently reflecting the specificity of the indigenous facilitated diffusion transport system. This microorganism may be useful, along with the previously developed xylose-fermenting Z. mobilis (M. Zhang, C. Eddy, K. Deanda, M. Finkelstein, and S. Picataggio, Science 267:240-243, 1995), in a mixed culture for efficient fermentation of the predominant hexose and pentose sugars in agricultural residues and other lignocellulosic feedstocks to ethanol.

摘要

产乙醇细菌运动发酵单胞菌的底物发酵范围得以扩展,纳入了戊糖L-阿拉伯糖,这种糖常见于农业废弃物和其他木质纤维素生物质中。从大肠杆菌中分离出五个基因,分别编码L-阿拉伯糖异构酶(araA)、L-核糖激酶(araB)、L-核糖ulose-5-磷酸-4-表异构酶(araD)、转醛醇酶(talB)和转酮醇酶(tktA),并在组成型启动子的控制下导入运动发酵单胞菌,这些启动子能使它们即使在有葡萄糖存在的情况下也能表达。基于消耗糖的量,工程菌株以L-阿拉伯糖作为唯一碳源生长并产生乙醇,乙醇产量达到最大理论产量的98%。这表明阿拉伯糖几乎完全代谢为乙醇这一唯一发酵产物,几乎没有副产物形成。尽管没有明显的二次生长模式,但该微生物在利用阿拉伯糖之前优先利用葡萄糖,这显然反映了原生促进扩散转运系统的特异性。这种微生物可能与之前开发的能发酵木糖的运动发酵单胞菌(M. Zhang、C. Eddy、K. Deanda、M. Finkelstein和S. Picataggio,《科学》267:240 - 243,1995年)一起,用于混合培养,以将农业废弃物和其他木质纤维素原料中主要的己糖和戊糖高效发酵为乙醇。

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