Efficient Production of the Dicarboxylic Acid Glutarate by Corynebacterium glutamicum via a Novel Synthetic Pathway.
Efficient Production of the Dicarboxylic Acid Glutarate by Corynebacterium glutamicum via a Novel Synthetic Pathway.
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谷氨酸棒杆菌通过一种新型合成途径高效生产戊二酸（二羧酸）
DOI：
10.3389/fmicb.2018.02589
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发表时间：
2018
影响因子：
5.2
通讯作者：
Wendisch VF
中科院分区：
文献类型：
--
作者：
Pérez-García F;Jorge JMP;Dreyszas A;Risse JM;Wendisch VF
戊二酸二羧酸是一种重要的结构单元，在化学和制药行业中引起了人们的兴趣。在此，开发了一种通过放线菌谷氨酸棒杆菌发酵生产戊二酸的合成途径。该途径不需要分子氧，通过赖氨酸脱羧酶进行操作，然后进行两个转氨基反应和两个 NAD 依赖性氧化反应。以基因组精简的L-赖氨酸生产菌株为基础，进行代谢工程，以五步合成途径将L-赖氨酸转化为戊二酸，该合成途径包括来自大肠杆菌的赖氨酸脱羧酶、腐胺转氨酶和γ-氨基丁醛脱氢酶以及来自谷氨酸棒杆菌或来自三种的GABA/5AVA氨基转移酶和琥珀酸/戊二酸半醛脱氢酶假单胞菌属物种。通过删除各自的乙酰化酶和输出基因，避免了由于副产物尸胺和N-乙酰尸胺的形成而造成的碳损失。由于合成戊二酸生物合成途径中的两个转氨反应产生 L-谷氨酸，因此通过谷氨酸脱氢酶进行 L-谷氨酸生物合成预计将被淘汰，事实上，删除其基因 gdh 可使戊二酸滴度增加 10%。在生物反应器（n = 2）中测试了最终菌株的戊二酸产量，以研究该过程的稳定性和可靠性。通过补料分批发酵 (n = 1) 实现了最有效的葡萄糖生产戊二酸，其体积生产率为 0.32 g L-1 h-1，总产量为 0.17 g g-1，效价为 25 g L-1。
The dicarboxylic acid glutarate is an important building-block gaining interest in the chemical and pharmaceutical industry. Here, a synthetic pathway for fermentative production of glutarate by the actinobacterium Corynebacterium glutamicum has been developed. The pathway does not require molecular oxygen and operates via lysine decarboyxylase followed by two transamination and two NAD-dependent oxidation reactions. Using a genome-streamlined L-lysine producing strain as basis, metabolic engineering was performed to enable conversion of L-lysine to glutarate in a five-step synthetic pathway comprising lysine decarboxylase, putrescine transaminase and γ-aminobutyraldehyde dehydrogenase from Escherichia coli and GABA/5AVA amino transferase and succinate/glutarate semialdehyde dehydrogenase either from C. glutamicum or from three Pseudomonas species. Loss of carbon via formation of the by-products cadaverine and N-acetylcadaverine was avoided by deletion of the respective acetylase and export genes. As the two transamination reactions in the synthetic glutarate biosynthesis pathway yield L-glutamate, biosynthesis of L-glutamate by glutamate dehydrogenase was expected to be obsolete and, indeed, deletion of its gene gdh increased glutarate titers by 10%. Glutarate production by the final strain was tested in bioreactors (n = 2) in order to investigate stability and reliability of the process. The most efficient glutarate production from glucose was achieved by fed-batch fermentation (n = 1) with a volumetric productivity of 0.32 g L-1 h-1, an overall yield of 0.17 g g-1 and a titer of 25 g L-1.
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影响因子：
4.2
作者：
Leßmeier L;Wendisch VF
通讯作者：
Wendisch VF
影响因子：
8.4
作者：
Becker, Judith;Zelder, Oskar;Wittmann, Christoph
通讯作者：
Wittmann, Christoph
影响因子：
3.8
作者：
Jorge, Joao M. P.;Nguyen, Anh Q. D.;Wendisch, Volker F.
通讯作者：
Wendisch, Volker F.
影响因子：
5
作者：
Lubitz, Dorit;Jorge, Joao M. P.;Wendisch, Volker F.
通讯作者：
Wendisch, Volker F.
影响因子：
4.4
作者：
Blombach, Bastian;Riester, Tanja;Eikmanns, Bernhard J.
通讯作者：
Eikmanns, Bernhard J.