构建pyc启动子突变文库改造谷氨酸棒杆菌高效生产β-丙氨酸
onstruct a pyc promoter library and modify Corynebacterium glutamicum to produce β-alanine efficiently
β-丙氨酸是唯一天然存在的β型氨基酸,在制药、食品和化学工业中有着广泛的应用。鉴于日益增长的市场需求,β-丙氨酸生产的研究是至关重要的。本研究利用改良的产赖氨酸菌株XQ-5.5作为出发菌株,通过底盘细胞以及pyc启动子突变文库的构建进一步促进β-丙氨酸的合成。为了增加细胞内草酰乙酸的供应并确保其转化为天冬氨酸(β-丙氨酸的直接前体),磷酸烯醇式丙酮酸羧化酶编码基因pck被删除。同时为了保证充足的丙酮酸维持三羧酸循环和草酰乙酸的合成,对丙酮酸氧化酶编码基因poxB也进行了删除。接下来,试图对基因pyc进行精细调控进一步保证充足的草酰乙酸含量供β-丙氨酸的合成。通过pyc启动子随机突变文库的构建与筛选提高丙酮酸到草酰乙酸的碳通量。最终,菌株XQ-5.7在5 L发酵罐中分批补料培养,β-丙氨酸产量为62.8 g L-1。
β-Alanine is the only naturally occurring β-type amino acid, with various applications in the pharmaceutical, food, and chemical industries. Given the growing market demand, the study of β-alanine production is important. In this study, the modified lysine-producing strain XQ-5.5 was used as the starting strain to further promote the synthesis of β-alanine through the construction of chassis cells and pyc promoter mutation library. To increase the intracellular supply of oxaloacetate and ensure its conversion to aspartate (a direct precursor of β-alanine), the phosphoenolpyruvate carboxylase coding gene pck was deleted. In order to ensure sufficient pyruvate to maintain tricarboxylic acid cycle and oxalacetate synthesis, the pyruvate oxidase coding gene poxB was also deleted. Next, an attempt was made to fine-regulate the gene pyc to further ensure sufficient oxalacetate content for β-alanine synthesis. The carbon flux from pyruvate to oxalacetate was increased through the construction and screening of pyc promoter random mutation library. Finally, the strain XQ-5.7 was cultured in a 5L fermenter with batch feeding, and the yield of β-alanine was 62.8 g L-1.
宋雅琪、张伟国、徐建中
生物工程学生物化学
β-丙氨酸谷氨酸棒杆菌代谢工程生物调控元件
orynebacterium glutamicumβ-alanineMetabolic engineeringBiological control elements.
宋雅琪,张伟国,徐建中.构建pyc启动子突变文库改造谷氨酸棒杆菌高效生产β-丙氨酸[EB/OL].(2024-03-06)[2025-08-11].http://www.paper.edu.cn/releasepaper/content/202403-70.点此复制
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