工程 Gluconobacter cerinus CGMCC 1.110 用于直接生产 2-酮基-L-古洛糖酸,Applied Microbiology and Biotechnology

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工程 Gluconobacter cerinus CGMCC 1.110 用于直接生产 2-酮基-L-古洛糖酸
Applied Microbiology and Biotechnology ( IF 3.9 ) Pub Date : 2022-11-29 , DOI: 10.1007/s00253-022-12310-5
Zhijie Qin _{1,

2,

3,

4} , Yue Chen _{1,

2,

3,

4} , Shiqin Yu _{1,

2,

3,

4} , Jian Chen _{1,

2,

3,

4} , Jingwen Zhou _{1,

2,

3,

4}

Affiliation

摘要

Gluconobacter是单步生产 2-keto-L-gulonic acid (2-KLG) 的潜在菌株，2-keto-L-gulonic acid (2-KLG) 是维生素 C 的直接前体。三种脱氢酶，即山梨糖醇脱氢酶 (SLDH)、山梨糖脱氢酶 (SDH)和山梨糖酮脱氢酶 (SNDH)，参与从 D-山梨糖醇生产 2-KLG。在本研究中，Gluconobacter cerinus菌株中潜在的 SNDH/SDH 基因簇通过基因组分析挖掘出CGMCC 1.110，验证了其将L-山梨糖转化为2-KLG的功能。蛋白质组学分析表明SNDH/SDH的表达水平对2-KLG的效价影响很大，发酵结果表明SDH是限速酶。然后进行系统的代谢工程过程，理论上适用于增加许多涉及来自Gluconobacter的膜结合脱氢酶的产品的滴度，以将G. cerinus CGMCC 1.110中的 2-KLG 滴度从检测不到提高到 51.9 g/L发酵优化后的 5 L 生物反应器。本研究采用的策略可为挖掘葡糖杆菌的其他潜在应用提供参考。

关键点

•挖掘了G. cerinus CGMCC 1.110 中潜在的SNDH/SDH 基因簇。

•执行了系统工程过程以提高2-KLG 的效价。

• 2-KLG 滴度成功地从检测不到增加到 51.9 g/L。

"点击查看英文标题和摘要"

Engineering Gluconobacter cerinus CGMCC 1.110 for direct 2-keto-L-gulonic acid production

Abstract

Gluconobacter is a potential strain for single-step production of 2-keto-L-gulonic acid (2-KLG), which is the direct precursor of vitamin C. Three dehydrogenases, namely, sorbitol dehydrogenase (SLDH), sorbose dehydrogenase (SDH), and sorbosone dehydrogenase (SNDH), are involved in the production of 2-KLG from D-sorbitol. In the present study, the potential SNDH/SDH gene cluster in the strain Gluconobacter cerinus CGMCC 1.110 was mined by genome analysis, and its function in transforming L-sorbose to 2-KLG was verified. Proteomic analysis showed that the expression level of SNDH/SDH had a great influence on the titer of 2-KLG, and fermentation results showed that SDH was the rate-limiting enzyme. A systematic metabolic engineering process, which was theoretically suitable for increasing the titer of many products involving membrane-bound dehydrogenase from Gluconobacter, was then performed to improve the 2-KLG titer in G. cerinus CGMCC 1.110 from undetectable to 51.9 g/L in a 5-L bioreactor after fermentation optimization. The strategies used in this study may provide a reference for mining other potential applications of Gluconobacter.