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构建并鉴定能够高效转运木糖的重组枯草芽孢杆菌 JY123。

Construction and characterization of recombinant Bacillus subtilis JY123 able to transport xylose efficiently.

机构信息

Department of Advanced Fermentation Fusion Science and Technology, Kookmin University, Seoul 136-702, Republic of Korea.

出版信息

J Biotechnol. 2012 Nov 15;161(4):402-6. doi: 10.1016/j.jbiotec.2012.07.192. Epub 2012 Aug 10.

Abstract

It has been known that wild type Bacillus subtilis cannot grow rapidly in a minimal medium containing xylose as a sole carbon source because it does not have a xylose-specific transporter. In this study, the arabinose:H(+) symporter, AraE protein from B. subtilis was expressed in B. subtilis 168 in order to transport xylose efficiently. The AraE expression cassette was constructed to contain the xylose-inducible xylA promoter, araE gene and fba terminator, and integrated into the chromosomal amyE gene in B. subtilis 168. Batch cultures in a defined medium with xylose only or a mixture of xylose and glucose showed that expression of AraE led to fast and complete consumption of initially added xylose and hence a considerable increase in cell growth of the recombinant B. subtilis JY123 expressing AraE. Considering the systematic analysis of cell growth, sugar consumption, respiratory quotient and xylulokinase activity, it was certain that AraE protein could transport xylose into B. subtilis efficiently.

摘要

已知野生型枯草芽孢杆菌不能在以木糖为唯一碳源的基础培养基中快速生长,因为它没有木糖特异性转运蛋白。在这项研究中,为了高效转运木糖,我们在枯草芽孢杆菌 168 中表达了阿拉伯糖:H(+)协同转运蛋白 AraE 蛋白。AraE 表达盒包含木糖诱导的 xylA 启动子、araE 基因和 fba 终止子,并整合到枯草芽孢杆菌 168 的 amyE 基因中。在含有木糖或木糖和葡萄糖混合物的限定培养基中的分批培养表明,AraE 的表达导致初始添加的木糖快速且完全消耗,从而使表达 AraE 的重组枯草芽孢杆菌 JY123 的细胞生长显著增加。考虑到对细胞生长、糖消耗、呼吸商和木酮糖激酶活性的系统分析,可以确定 AraE 蛋白可以有效地将木糖转运到枯草芽孢杆菌中。

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