menA过表达菌株构建及两阶段pH控制促进VK2合成
Enhance Vitamin K2 Production Via menA Gene Overexpression and a Two-Stage pH Control Strategy
DOI:10.3969/j.issn.1673-1689.2019.12.005
中文关键词: 纳豆芽孢杆菌 过表达 控制策略 pH 维生素K2
英文关键词: Bacillus subtilisnatto,overexpression,control strategy,pH,vitamin K2
基金项目:
作者
单位
刘艳
安徽工程大学 生物与化学工程学院,安徽 芜湖 241000
芜湖张恒春药业有限公司,安徽 芜湖 241009
杨自名
薛正莲
杨超英
胡刘秀
程良成
王洲
丁秀敏
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中文摘要:
为提高纳豆芽孢杆菌合成维生素K2的能力,构建了纳豆芽孢杆菌体内维生素K2合成途径中的关键酶—1,4-二羟基-2-萘甲酸-聚异戊二烯转移酶(MenA)—过表达菌株,并在5 L发酵罐水平考察了不同pH对该过表达菌株生长和维生素K2合成能力的影响。结果表明:menA基因的过量表达能使Bacillus subtilis natto维生素K2合成量提高51%。单一恒定的pH不能满足既能使菌株生长充分,又能最大化产物合成的条件。通过分析不同pH下发酵过程曲线和动力学参数,提出了两阶段pH控制策略:即在发酵前期0~40 h控制pH 7.0,发酵后期40~80 h控制pH 4.0,菌体生物量和维生素K2合成量分别达到了(11.46±0.67) g/L和(63.60±1.81) mg/L,是原始菌分批发酵pH自然状态下的1.5倍和2.6倍。
英文摘要:
To improve vitamin K2 production,1,4-dihydroxy-2-naphthoicacid polyprenyltransferase (MenA)which was a key enzyme for vitamin K2synthesis inBacillus subtilisnatto was overexpressed. The effects of pH on cell growth and vitamin K2 production was also investigated in a 5 L fermentator. It was shown that the vitamin K2production was enhanced by 51% when menA gene was overexpressed. The mono-control of pH could not balance the cell growth and vitamin K2production. A two-stage pH control strategy was employed,i.e. pH was controlled at 7.0 between 0~40 h and switched to 4.0 between 40~80 h. The biomass and vitamin K2production reached(11.46±0.67)g/L and(63.60±1.81) mg/L,which were 1.5 and 2.6 times of that of pH uncontrolled condition by original strain,respectively.
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