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缺乏磷酸烯醇丙酮酸:碳水化合物磷酸转移酶系统的大肠杆菌菌株中的乙酸代谢;碳循环策略和无效循环的证据

Acetate metabolism in Escherichia coli strains lacking phosphoenolpyruvate: carbohydrate phosphotransferase system; evidence of carbon recycling strategies and futile cycles.

作者信息

Sigala Juan Carlos, Flores Salvador, Flores Noemí, Aguilar César, de Anda Ramón, Gosset Guillermo, Bolívar Francisco

机构信息

Departamento de Ingeniería Celular y Biocatálisis, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca/Morelos, México.

出版信息

J Mol Microbiol Biotechnol. 2009;16(3-4):224-35. doi: 10.1159/000151219. Epub 2008 Aug 5.

DOI:10.1159/000151219
PMID:18679018
Abstract

The ptsHIcrr operon was deleted from Escherichia coli wild-type JM101 to generate strain PB11 (PTS(-)). In a mutant derived from PB11 that partially recovered its growth capacity on glucose by an adaptive evolution process (PB12, PTS(-)Glc(+)), part of the phosphoenolpyruvate not used in glucose transport has been utilized for the synthesis of aromatic compounds. In this report, it is shown that on acetate as a carbon source, PB11 displayed a specific growth rate (mu) higher than PB12 (0.21 and 0.13 h(-1), respectively) while JM101 had a mu of 0.28 h(-1). To understand these growth differences on acetate, we compared the expression profiles of central metabolic genes by RT-PCR analysis. Obtained data revealed that some gluconeogenic genes were downregulated in both PTS(-) strains as compared to JM101, while most glycolytic genes were upregulated in PB12 in contrast to PB11 and JM101. Furthermore, inactivation of gluconeogenic genes, like ppsA, sfcA, and maeB,and poxB gene that codes for pyruvate oxidase, has differential impacts in the acetate metabolism of these strains. Results indicate that growth differences on acetate in the PTS(-) derivatives are due to potential carbon recycling strategies, mainly in PB11, and futile carbon cycles, especially in PB12.

摘要

将磷酸转移酶系统(PTS)的ptsHIcrr操纵子从大肠杆菌野生型JM101中删除,以构建PB11菌株(PTS缺失型)。在由PB11衍生的一个突变体中,该突变体通过适应性进化过程部分恢复了在葡萄糖上的生长能力(PB12,PTS缺失型葡萄糖利用能力恢复型),部分未用于葡萄糖转运的磷酸烯醇丙酮酸已被用于芳香族化合物的合成。在本报告中,研究表明,以乙酸盐作为碳源时,PB11的比生长速率(μ)高于PB12(分别为0.21和0.13 h⁻¹),而JM101的μ为0.28 h⁻¹。为了解这些菌株在乙酸盐上的生长差异,我们通过逆转录聚合酶链反应(RT-PCR)分析比较了中心代谢基因的表达谱。获得的数据显示,与JM101相比,两个PTS缺失型菌株中的一些糖异生基因表达下调,而与PB11和JM101相比,大多数糖酵解基因在PB12中表达上调。此外,糖异生基因(如ppsA、sfcA和maeB)以及编码丙酮酸氧化酶的poxB基因的失活,对这些菌株的乙酸盐代谢有不同影响。结果表明,PTS缺失型衍生物在乙酸盐上的生长差异是由于潜在的碳循环策略(主要在PB11中)和无效的碳循环(特别是在PB12中)。

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