缺失分枝菌酸层提高谷氨酸棒状杆菌L-谷氨酸产量的研究
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国家“十四五”重点研发计划项目(2021YFC2100900)


Deletion of Mycolic Acid Layer Increased L-glutamate Production in Corynebacterium glutamicum
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    摘要:

    谷氨酸棒状杆菌(Corynebacterium glutamicumC. glutamicum)的分枝菌酸层对维持细胞的通透性有重要作用,但是其合成需要消耗大量的底物和能源。作者通过敲除谷氨酸棒状杆菌ATCC13869中聚酮合酶编码基因pks13,构建了突变菌株Δpks13。与野生型相比,突变菌Δpks13细胞体积变大、形状变圆且细胞壁结合松散容易脱落,但L-谷氨酸产量提高了8倍。薄层层析和液相色谱-质谱联用分析发现,Δpks13菌株细胞壁中的脂质主要为磷脂。实时荧光定量PCR分析发现,与野生型相比,Δpks13谷氨酸外排相关基因mscCG的转录水平上调,而α-酮戊二酸脱氢酶编码基因odhA和下游产物精氨酸合成相关基因(argBargCargDargFargGargHargJ)转录水平下调。这些基因转录水平的改变解释了Δpks13 L-谷氨酸产量提高的机制。

    Abstract:

    The mycolic acid layer in Corynebacterium glutamicum (C. glutamicum) plays important role in maintain cell permeability, however, its synthesis consumes substantial energy and substrate in the synthesis process. A mutant strain, Δpks13, was constructed by deleting the polyketide synthase gene pks13 in C. glutamicum ATCC13869. Compared to the wild type, the mutant strain Δpks13 cells were larger, more rounded, and with a loosely bound cell wall that tended to detach easily,while the L-glutamate yield increased 8-fold. Thin layer chromatography and liquid chromatography mass spectrometry analysis revealed that phospholipids were the primary cell wall lipids in Δpks13. Quantitative real-time PCR analysis revealed that, compared with wild-type strain,the transcript level of the glutamate efflux-related gene mscCG was upregulated, while the transcription levels of the α-ketoglutarate dehydrogenase-encoding gene odhA and the downstream arginine synthesis-related gene (argB, argC, argD, argF, argG, argH and argJ) were downregulated in Δpks13 cells. These changes in genes transcription levels explain the mechanism underlying the enhanced L-glutamate production in Δpks13.

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李贺丹,张德志,赵桂红,李姿含,胡晓清,王小元.缺失分枝菌酸层提高谷氨酸棒状杆菌L-谷氨酸产量的研究[J].食品与生物技术学报,2024,43(8):31-40.

LI He-dan, ZHANG De-zhi, ZHAO Gui-hong, LI Zi-han, HU Xiao-qing, WANG Xiao-yuan. Deletion of Mycolic Acid Layer Increased L-glutamate Production in Corynebacterium glutamicum[J]. Journal of Food Science and Biotechnology,2024,43(8):31-40.

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  • 在线发布日期: 2024-12-11
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