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本文引用的文献

1
Modelling overflow metabolism in Escherichia coli with flux balance analysis incorporating differential proteomic efficiencies of energy pathways.利用通量平衡分析对大肠杆菌中的溢流代谢进行建模，该分析纳入了能量途径的差异蛋白质组学效率。
BMC Syst Biol. 2019 Jan 10;13(1):3. doi: 10.1186/s12918-018-0677-4.
2
Enhancement of NAD(H) pool for formation of oxidized biochemicals in Escherichia coli.增强 NAD(H) 池以在大肠杆菌中形成氧化生物化学物质。
J Ind Microbiol Biotechnol. 2018 Nov;45(11):939-950. doi: 10.1007/s10295-018-2072-y. Epub 2018 Aug 29.
3
Improving E. coli growth performance by manipulating small RNA expression.通过操纵小 RNA 表达来提高大肠杆菌的生长性能。
Microb Cell Fact. 2017 Nov 14;16(1):198. doi: 10.1186/s12934-017-0810-x.
4
Overflow metabolism in Escherichia coli results from efficient proteome allocation.大肠杆菌中的代谢溢出是由于有效的蛋白质组分配所致。
Nature. 2015 Dec 3;528(7580):99-104. doi: 10.1038/nature15765.
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Constitutive expression of the sRNA GadY decreases acetate production and improves E. coli growth.小RNA GadY的组成型表达可减少乙酸盐生成并改善大肠杆菌的生长。
Microb Cell Fact. 2015 Sep 18;14:148. doi: 10.1186/s12934-015-0334-1.
6
Reduction of Acetate and Lactate Contributed to Enhancement of a Recombinant Protein Production in E. coli BL21.乙酸盐和乳酸盐的减少有助于提高大肠杆菌BL21中重组蛋白的产量。
J Microbiol Biotechnol. 2015 Jul;25(7):1093-100. doi: 10.4014/jmb.1503.03023.
7
Aerobic expression of Vitreoscilla hemoglobin efficiently reduces overflow metabolism in Escherichia coli.透明颤菌血红蛋白的需氧表达有效降低了大肠杆菌中的溢流代谢。
Biotechnol J. 2014 Jun;9(6):791-9. doi: 10.1002/biot.201300388. Epub 2014 Mar 18.
8
Systems biology approach reveals that overflow metabolism of acetate in Escherichia coli is triggered by carbon catabolite repression of acetyl-CoA synthetase.系统生物学方法表明，大肠杆菌中乙酸盐的溢流代谢是由乙酰辅酶A合成酶的碳分解代谢物阻遏引发的。
BMC Syst Biol. 2010 Dec 1;4:166. doi: 10.1186/1752-0509-4-166.
9
Acetate accumulation through alternative metabolic pathways in ackA (-) pta (-) poxB (-) triple mutant in E. coli B (BL21).在大肠杆菌 B (BL21) 的 ackA (-) pta (-) poxB (-) 三重突变体中，通过替代代谢途径积累醋酸盐。
Biotechnol Lett. 2010 Dec;32(12):1897-903. doi: 10.1007/s10529-010-0369-7. Epub 2010 Aug 12.
10
Metabolic selective pressure stabilizes plasmids carrying biosynthetic genes for reduced biochemicals in Escherichia coli redox mutants.代谢选择压力稳定了携带生物合成基因的质粒，这些基因用于减少大肠杆菌氧化还原突变体中的生化物质。
Appl Microbiol Biotechnol. 2010 Sep;88(2):563-73. doi: 10.1007/s00253-010-2774-1. Epub 2010 Jul 30.

在具有升高的NAD（H）库的K-12中重组蛋白生产过程中的乙酸盐形成。

Acetate formation during recombinant protein production in K-12 with an elevated NAD(H) pool.

作者信息

Han Qi, Eiteman Mark A

机构信息

School of Chemical Materials and Biomedical Engineering University of Georgia Athens GA USA.

出版信息

Eng Life Sci. 2019 Sep 8;19(11):770-780. doi: 10.1002/elsc.201900045. eCollection 2019 Nov.

DOI:10.1002/elsc.201900045
PMID:32624970
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6999358/
Abstract

Acetate formation is a disadvantage in the use of for recombinant protein production, and many studies have focused on optimizing fermentation processes or altering metabolism to eliminate acetate accumulation. In this study, MEC697 (MG1655 ) maintained a larger pool of NAD(H) compared to the wild-type control, and also accumulated lower concentrations of acetate when grown in batch culture on glucose. In steady-state cultures, the elevated total NAD(H) found in MEC697 delayed the threshold dilution rate for acetate formation to a growth rate of 0.27 h. Batch and fed-batch processes using MEC697 were examined for the production of β-galactosidase as a model recombinant protein. Fed-batch culture of MEC697/pTrc99A- compared to MG1655/pTrc99A- at a growth rate of 0.22 h showed only a modest increase of protein formation. However, 1 L batch growth of MEC697/pTrc99A- resulted in 50% lower acetate formation compared to MG1655/pTrc99A- and a two-fold increase in recombinant protein production.

摘要

在用于重组蛋白生产时，乙酸盐的形成是一个不利因素，许多研究都集中在优化发酵过程或改变代谢以消除乙酸盐积累。在本研究中，与野生型对照相比，MEC697（MG1655）维持了更大的NAD（H）库，并且在以葡萄糖进行分批培养时，乙酸盐的积累浓度也更低。在稳态培养中，MEC697中升高的总NAD（H）将乙酸盐形成的阈值稀释率延迟至0.27 h的生长速率。使用MEC697的分批和补料分批工艺用于生产作为模型重组蛋白的β-半乳糖苷酶。在0.22 h的生长速率下，与MG1655/pTrc99A-相比，MEC697/pTrc99A-的补料分批培养仅显示蛋白质形成有适度增加。然而，与MG1655/pTrc99A-相比，MEC697/pTrc99A-的1 L分批培养导致乙酸盐形成降低50%，重组蛋白产量增加两倍。