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在[具体内容缺失]中开发基于质粒的表达系统及其用于筛选双功能醇脱氢酶的异源表达。

Development of a plasmid-based expression system in and its use to screen heterologous expression of bifunctional alcohol dehydrogenases (s).

作者信息

Hon Shuen, Lanahan Anthony A, Tian Liang, Giannone Richard J, Hettich Robert L, Olson Daniel G, Lynd Lee R

机构信息

Thayer School of Engineering, Dartmouth College, Hanover, NH, USA.

BioEnergy Science Center, Oak Ridge, TN, USA.

出版信息

Metab Eng Commun. 2016 Apr 22;3:120-129. doi: 10.1016/j.meteno.2016.04.001. eCollection 2016 Dec.

DOI:10.1016/j.meteno.2016.04.001
PMID:29142822
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5678826/
Abstract

is a promising candidate for ethanol production from cellulosic biomass, but requires metabolic engineering to improve ethanol yield. A key gene in the ethanol production pathway is the bifunctional aldehyde and alcohol dehydrogenase, . To explore the effects of overexpressing wild-type, mutant, and exogenous s, we developed a new expression plasmid, pDGO144, that exhibited improved transformation efficiency and better gene expression than its predecessor, pDGO-66. This new expression plasmid will allow for many other metabolic engineering and basic research efforts in . As proof of concept, we used this plasmid to express 12 different genes (both wild type and mutant) from several organisms. Ethanol production varied between clones immediately after transformation, but tended to converge to a single value after several rounds of serial transfer. The previously described mutant D494G gave the best ethanol production, which is consistent with previously published results.

摘要

是纤维素生物质生产乙醇的一个有潜力的候选者，但需要代谢工程来提高乙醇产量。乙醇生产途径中的一个关键基因是双功能醛和醇脱氢酶，。为了探究过表达野生型、突变型和外源的影响，我们开发了一种新的表达质粒pDGO144，它比其前身pDGO - 66表现出更高的转化效率和更好的基因表达。这种新的表达质粒将有助于在中开展许多其他代谢工程和基础研究工作。作为概念验证，我们使用该质粒表达了来自几种生物体的12种不同的基因（野生型和突变型）。转化后立即克隆间的乙醇产量有所不同，但经过几轮连续传代后趋于收敛到一个单一值。先前描述的突变型D494G产生了最佳的乙醇产量，这与先前发表的结果一致。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e227/5678826/b275c7c5fa18/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e227/5678826/5019b9619c18/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e227/5678826/0d6f9bb96ef8/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e227/5678826/87472013ed62/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e227/5678826/7bff31f833f8/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e227/5678826/b275c7c5fa18/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e227/5678826/5019b9619c18/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e227/5678826/0d6f9bb96ef8/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e227/5678826/87472013ed62/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e227/5678826/7bff31f833f8/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e227/5678826/b275c7c5fa18/gr5.jpg

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