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通过异源表达 L-乳酸脱氢酶基因在柠檬明串珠菌中生产 L-乳酸。

Production of L-lactate in Leuconostoc citreum via heterologous expression of L-lactate dehydrogenase gene.

机构信息

Department of Food Science and Technology, Research Center for Bioresource and Health, Chungbuk National University, Cheongju, South Korea.

出版信息

J Biotechnol. 2009 Oct 26;144(2):160-4. doi: 10.1016/j.jbiotec.2009.08.012. Epub 2009 Aug 21.

Abstract

D-form lactate is often found in fermented foods and excessive dietary intake of D-lactate may cause metabolic stress in both infants and patients. Leuconostoc citreum is a major lactic acid bacterium that produces D-lactate in fermented foods. The aim of this study was to change the pyruvate carbon flux in L. citreum from D-lactate into L-lactate by heterologous expression of L-lactate dehydrogenase (ldhL) gene. For this, ldhL from Lactobacillus plantarum was cloned and introduced into L. citreum using a shuttle vector pLeuCM. In the transformant, ldhL was successfully transcribed and L-lactate dehydrogenase was expressed. As a consequence of transformation, the ratio between D- and L-isomers was changed due to the increment of L-lactate and the decrement of D-lactate, but no significant differences were found in total lactate concentration between the host and transformant cells. This is the first report of metabolic engineering in Leuconostoc by modulating the central carbon flux into health-favored way.

摘要

D-乳酸通常存在于发酵食品中,过量摄入 D-乳酸可能会给婴儿和患者带来代谢应激。肠膜明串珠菌是一种主要的乳酸细菌,它在发酵食品中产生 D-乳酸。本研究旨在通过异源表达 L-乳酸脱氢酶(ldhL)基因将 L. citreum 中的丙酮酸碳通量从 D-乳酸转变为 L-乳酸。为此,从植物乳杆菌中克隆了 ldhL 并使用穿梭载体 pLeuCM 引入到 L. citreum 中。在转化体中,ldhL 成功转录并表达了 L-乳酸脱氢酶。由于转化,由于 L-乳酸的增加和 D-乳酸的减少,D-和 L-异构体的比例发生了变化,但在宿主和转化细胞之间的总乳酸浓度没有发现显著差异。这是通过调节中心碳通量以有利于健康的方式对肠膜明串珠菌进行代谢工程的首次报道。

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