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利用葡萄糖和替代碳源,通过谷氨酸棒杆菌提高相容溶质海藻糖的发酵生产。

Improved fermentative production of the compatible solute ectoine by Corynebacterium glutamicum from glucose and alternative carbon sources.

机构信息

Genetics of Prokaryotes, Faculty of Biology & CeBiTec, Bielefeld University, Universitätsstr. 25, 33615 Bielefeld, Germany.

Fermentation Technology, Technical Faculty & CeBiTec, Bielefeld University, Universitätsstr. 25, 33615 Bielefeld, Germany.

出版信息

J Biotechnol. 2017 Sep 20;258:59-68. doi: 10.1016/j.jbiotec.2017.04.039. Epub 2017 May 3.

Abstract

The cyclic amino acid ectoine is a compatible solute serving as a protective substance against osmotic stress. Ectoine finds various applications due to its moisturizing effect. To avoid the disadvantages of the prevailing so-called "bacterial milking ectoine production process" caused by the high salt concentration, low salt fermentation strategies are sought after. As l-lysine and ectoine biosynthesis share l-aspartate-semialdehyde as common precursor, l-lysine producing strains can be converted to ectoine producing strains. Corynebacterium glutamicum, which is used for l-lysine production in the million-ton-scale, was engineered for ectoine production by heterologous expression of the ectoine biosynthesis operon ectABC from Chromohalobacter salexigens. Derepression of glucose metabolism by deletion of the regulatory gene sugR and avoiding l-lactate formation by deletion of the lactate dehydrogenase gene ldhA increased ectoine productivity. In bioreactor fed-batch cultivations an ectoine titer of 22gL and a volumetric productivity of 0.32gLh were obtained. The ectoine yield of 0.16gg, to the best of our knowledge, exceeded previously reported yields. Moreover, ectoine production from the alternative carbon sources glycerol, glucosamine, xylose, arabinose, and soluble starch was achieved.

摘要

环状氨基酸羟乙基呱嗪乙酸盐是一种可与水混溶的溶质,作为一种抗渗透胁迫的保护物质。由于其保湿作用,羟乙基呱嗪乙酸盐有多种应用。为了避免由于高盐浓度而导致的现有所谓“细菌挤奶羟乙基呱嗪生产工艺”的缺点,人们寻求低盐发酵策略。由于 l-赖氨酸和羟乙基呱嗪生物合成共享 l-天冬氨酸-4-半醛醛作为共同前体,因此可以将生产 l-赖氨酸的菌株转化为生产羟乙基呱嗪的菌株。用于百万吨级 l-赖氨酸生产的谷氨酸棒杆菌通过异源表达来自盐红杆菌的羟乙基呱嗪生物合成操纵子 ectABC 进行了羟乙基呱嗪生产工程改造。通过删除调节基因 sugR 来解除葡萄糖代谢的阻遏作用,以及通过删除乳酸脱氢酶基因 ldhA 来避免形成 l-乳酸,提高了羟乙基呱嗪的生产力。在生物反应器分批补料培养中,获得了 22gL 的羟乙基呱嗪浓度和 0.32gLh 的比生产率。据我们所知,羟乙基呱嗪的产率为 0.16gg,超过了之前报道的产率。此外,还实现了从替代碳源甘油、葡糖胺、木糖、阿拉伯糖和可溶性淀粉生产羟乙基呱嗪。

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