通过催化假定的托芬合成蛋白酶的托芬生产
Lulu Cao1, Jiaqi Zhang1,2, Jia Chen1
1Cooperative Innovation Center of Industrial Fermentation (Ministry of Education & Hubei Province), Key Laboratory of Fermentation Engineering (Ministry of Education), National "111", Center for Cellular Regulation and Molecular Pharmaceutics, Hubei University of Technology, Wuhan, 430068, PR China.
Archives of microbiology
|September 2, 2024
概括
氨酸合成酶是生产氨酸的必不可少的酶,已经成功地从Sphingomonas soilsilvae克隆并表达出来. 这项研究优化了它的活性,并确定了功能性食品中潜在的大规模托合成的关键活性部位.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 食品科学 食品科学 食品科学
背景情况:
- 托芬是一种必需的氨基酸,对代谢功能和功能性食物发育至关重要.
- 在环保细菌中发现了一种托合成酶基因 Sphingobacterium soilsilvae Em02.02.
研究的目的:
- 克隆和表达来自S. soilsilvae在E. coli中的托芬合成酶基因.
- 描述酶的活性,最佳条件和活性位点.
主要方法:
- 在大肠杆菌中基因克隆和基因表达.
- 使用印和L-氨酸的酶活性测定.
- 部位定向突变发生,以确定活性部位.
- 优化反应条件 (温度,pH,金属离子).
主要成果:
- 再组合三合成酶 (42 KDa) 在可溶性形式成功表达.
- 在50°C和pH值7.0下观察到最佳活性,增强了Mg2+,Sr2+和Mn2+.
- 在共识模式HK-S-[GGGSN]-E-S中的关键活性位点通过突变发生法被确定.
结论:
- 有特征的托合成酶表现出β子单元的特性.
- 这种酶具有大规模生产托的潜力.
- 食品科学中的应用可以提高食品的营养含量和健康益处.
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