溶解氧对γ-聚谷氨酸合成的影响

doi:10.13523/j.cb.20150306

中国生物工程杂志

2015, Vol. 35

Issue (3): 42-48 DOI: 10.13523/j.cb.20150306

研究报告

溶解氧对γ-聚谷氨酸合成的影响

李小鑫¹, 高明昊¹, 张苗苗¹, 刘晓晨¹, 乔长晟^1,2

1. 天津科技大学工业发酵微生物教育部重点实验室天津 300457;
2. 天津北洋百川生物技术有限公司天津 300457

Effect of Dissolved Oxygen on γ-PGA Fermentation

LI Xiao-xin¹, GAO Ming-hao¹, ZHANG Miao-miao¹, LIU Xiao-chen¹, QIAO Chang-sheng^1,2

1. Key Laboratory of Industrial Microbiology, Ministry of Education, Tianjin University of Science and Technology, Tianjin 300457, China;
2. Tianjin Peiyang Biotrans Company Limited, Tianjin 300457, China

全文: PDF(737 KB) HTML

摘要：

在5L发酵罐上研究了溶解氧(DO)对地衣芽孢杆菌分批发酵生产γ-聚谷氨酸(γ-PGA)的影响并考察在8h、32h、56h时,葡萄糖激酶、6-磷酸葡萄糖脱氢酶、丙酮酸脱氢酶、异柠檬酸脱氢酶和谷氨酸脱氢酶的活性及对应时间点上γ-PGA的生产速率.通过溶解氧电极和搅拌转速的串联控制发酵过程中溶解氧水平,发现高溶解氧(60%)水平和低溶解氧(10%)水平均不能高效积累γ-PGA.6-磷酸葡萄糖脱氢酶活性的提高对产物的积累有抑制作用,葡萄糖激酶和谷氨酸脱氢酶的酶活提高对产物积累有促进作用,过高的丙酮酸脱氢酶和异柠檬酸脱氢酶的活性在一定程度上可以促进菌体生长但不利于产物的积累.此外,通过对三种不同DO水平下γ-PGA生物合成途径中相关代谢流量的计算表明,在pH 6.5的条件下,对于谷氨酸依赖型生产菌株,提高外源谷氨酸利用率可以促进γ-PGA的生物合成.

关键词： γ -聚谷氨酸; 溶解氧; 代谢通量; 关键酶活性

Abstract:

Effects of different dissolved oxygen (DO) on the production rate of γ-PGA, at the 8, 32 and 56 hours were studied, using the 5L fermentor to synthesis γ-PGA with Bacillus licheniformis. Results indicated that neither high nor low DO value could efficiently accumulate γ-PGA based on the analysis of relevant data. The research also concerned about five kinds of enzyme activity which were key enzymes of metabolic flux of biosynthesis for γ-PGA. The enzymes includes hexokinase, glucose-6-phosphate dehydrogenase(G-6-Dp), pyruvate dehydrogenase(PDH), isocitrate dehydrogenase(ICDH)and glutamate dehydrogenase(GDH). And G-6-D Pcould depress the synthesis of γ-PGA. High activity of Hexokinase and GDH would promote the production of γ-PGA. PDH and ICDH might advance the production rate of cell. The higher activity of PDH and ICDH might be not conductive to the fermentation. Moreover, the extracellular metabolites profiles of fermentation under three different DO values were acquired and the metabolic flux redistribution of pathways related to γ-PGA biosynthesis was calculated based on the collected data. As a result, the metabolic flux favored to distribute toward glycolytic pathway at DO 30%, in which the ingestion rate of extracellular glutamic acid was higher and the subsequent γ-PGA biosynthesis was enhanced.

Key words: γ-PGA Dissolved oxygen Metabolic flux Key enzyme activity

收稿日期: 2014-10-08 出版日期: 2015-03-25

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通讯作者: 乔长晟 E-mail: qiaochangsheng@163.com

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引用本文:

李小鑫, 高明昊, 张苗苗, 刘晓晨, 乔长晟. 溶解氧对γ-聚谷氨酸合成的影响[J]. 中国生物工程杂志, 2015, 35(3): 42-48.

LI Xiao-xin, GAO Ming-hao, ZHANG Miao-miao, LIU Xiao-chen, QIAO Chang-sheng. Effect of Dissolved Oxygen on γ-PGA Fermentation. China Biotechnology, 2015, 35(3): 42-48.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20150306 或 https://manu60.magtech.com.cn/biotech/CN/Y2015/V35/I3/42

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