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首页> 美国卫生研究院文献>other >Heterologous Expression of Argininosuccinate Synthase From Oenococcus oeni Enhances the Acid Resistance of Lactobacillus plantarum
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Heterologous Expression of Argininosuccinate Synthase From Oenococcus oeni Enhances the Acid Resistance of Lactobacillus plantarum

机译：Oenococcus oeni的精氨酸琥珀酸合酶的异源表达增强了植物乳杆菌的耐酸性

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摘要

Oenococcus oeni can survive well in wine (an acid-stress environment) and dominate malolactic fermentation (MLF). To demonstrate a possible role of argininosuccinate synthase gene (argG) in the acid tolerance response of O. oeni, a related argG gene was inserted into a plasmid pMG36e and heterologously expressed in Lactobacillus plantarum SL09, a wine isolate belonging to a species of relevant importance in MLF. The expression levels of the argG gene in L. plantarum were analyzed by RT-qPCR, argininosuccinate synthase (ASS) activity and cell properties (amino acids, pH, H+-ATPase activity, and ATP levels) were determined at pH 3.7 in comparison with that at pH 6.3. Results showed that the recombinant strain L. plantarum SL09 (pMG36eargG) exhibited stronger growth performance compared with the control strain (without argG gene), and the expression levels of hsp1, cfa, atp, the citrate and malate metabolic genes were apparently increased under acid stress. In addition, the recombinant strain exhibited 11.0-, 2.0-, 1.9-fold higher ASS activity, H+-ATPase activity and intracellular ATP level, compared with the corresponding values for control strain during acid-stresses condition, which may take responsible for the acid tolerance enhancement of the recombinant strain. This is the first work report on heterologous expression of argG gene, and the results presented in this study will be beneficial for the research on acid stress response of O. oeni.
机译：Oenococcus oeni可以在葡萄酒（酸胁迫的环境）中很好地生存，并在苹果乳酸发酵（MLF）中占主导地位。为了证明精氨酸琥珀酸合酶基因（argG）在O. oeni的耐酸反应中的可能作用，将一个相关的argG基因插入质粒pMG36e中，并在植物乳杆菌SL09（一种具有重要意义的葡萄酒分离株）中异源表达。在MLF中。通过RT-qPCR，精氨酸琥珀酸合酶（ASS）活性和细胞特性（氨基酸，pH，H + -ATPase活性和ATP水平）分析了植物乳杆菌argG基因的表达水平。与pH 6.3相比，在pH 3.7下测定了pH值。结果表明，重组菌株植物乳杆菌SL09（pMG36eargG）比对照菌株（无argG基因）表现出更强的生长性能，在酸性条件下，hsp1，cfa，atp，柠檬酸和苹果酸代谢基因的表达水平明显升高。强调。此外，与酸胁迫条件下对照菌株的相应值相比，该重组菌株的ASS活性，H + -ATPase活性和细胞内ATP水平高出11.0-，2.0-，1.9倍。 ，这可能是重组菌株耐酸性增强的原因。这是关于argG基因异源表达的第一份工作报告，本研究结果将有益于O. oeni的酸胁迫反应研究。

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