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从葡萄糖和替代碳源发酵生产 5-氨基戊酸的新代谢途径。

A new metabolic route for the fermentative production of 5-aminovalerate from glucose and alternative carbon sources.

机构信息

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

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

出版信息

Bioresour Technol. 2017 Dec;245(Pt B):1701-1709. doi: 10.1016/j.biortech.2017.04.108. Epub 2017 Apr 29.

Abstract

Here, a new metabolic pathway for the production of 5-aminovalerate (5AVA) from l-lysine via cadaverine as intermediate was established and this three-step-pathway comprises l-lysine decarboxylase (LdcC), putrescine transaminase (PatA) and γ-aminobutyraldehyde dehydrogenase (PatD). Since Corynebacterium glutamicum is used for industrial l-lysine production, the pathway was established in this bacterium. Upon expression of ldcC, patA and patD from Escherichia coli in C. glutamicum wild type, production 5AVA was achieved. Enzyme assays revealed that PatA and PatD also converted cadaverine to 5AVA. Eliminating the by-products cadaverine, N-acetylcadaverine and glutarate in a genome-streamlined l-lysine producing strain expressing ldcC, patA and patD improved 5AVA production to a titer of 5.1gL, a yield of 0.13gg and a volumetric productivity of 0.12gLh. Moreover, 5AVA production from the alternative feedstocks starch, glucosamine, xylose and arabinose was established.

摘要

在这里,建立了一条从 l-赖氨酸经尸胺作为中间体生产 5-氨基戊酸(5AVA)的新代谢途径,该途径由 l-赖氨酸脱羧酶(LdcC)、腐胺转氨酶(PatA)和 γ-氨基丁醛脱氢酶(PatD)组成。由于谷氨酸棒状杆菌被用于工业 l-赖氨酸生产,因此该途径在该细菌中建立。在谷氨酸棒状杆菌野生型中表达来自大肠杆菌的 ldcC、patA 和 patD 后,实现了 5AVA 的生产。酶活性测定表明 PatA 和 PatD 也可将尸胺转化为 5AVA。在表达 ldcC、patA 和 patD 的基因组简化的 l-赖氨酸生产菌株中消除了副产物尸胺、N-乙酰尸胺和戊二酸,将 5AVA 的产量提高到 5.1gL、0.13gg 和 0.12gLh。此外,还建立了从替代原料淀粉、氨基葡萄糖、木糖和阿拉伯糖生产 5AVA 的途径。

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