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在Bacillus subtilis168中基于Pylb的P450细胞染色体过度表达

Thanaporn Wichai¹, Sarintip Sooksai¹, Sajee Noitang¹

¹Institute of Biotechnology and Genetic Engineering, Chulalongkorn University, Bangkok 10330, Thailand.

Enzyme and microbial technology

|January 26, 2025

概括

使用Bacillus subtilis中的Pylb促进体的无诱导体表达系统,可以为生物炼油应用提供高效的P450细胞染色体生产. 这种系统通过消除对化学诱导剂的需求来降低成本和简化过程.

科学领域:

生物技术是生物技术.
合成生物学合成生物学
工业微生物学工业微生物学

背景情况:

无诱导体的表达系统为生物炼油厂提供了成本效益和简化的生物工艺.
细胞染色体P450酶是生产增值化合物的多功能生物催化剂,但需要控制表达以避免细胞应激.

研究的目的:

评估来自Bacillus subtilis168的Pylb促进剂,用于开发B. subtilis中无诱导体表达系统.
为了评估Pylb促进剂在表达细胞染色体P450酶的有效性,用于生物炼油应用.

主要方法:

使用绿色光蛋白 (GFP) 记者来比较Pylb与重组大肠杆菌和B. subtilis.中的组成性促进物 (P43)
在30°C和37°C培养重组B. subtilis以确定最佳表达温度.
采用子优化的P450-BM3作为模型酶和紫色试验来确认酶表达和活性.

主要成果:

与P43相比,Pylb在大肠杆菌和B. subtilis中显示出不同的表达模式.
与30°C相比,在37°C的培养导致细菌光度增加2.8倍.
基于Pylb的系统成功地表达了活性细胞染色体P450-BM3,将八转化为八醇 (0.15 ± 0.04 mM 1-八醇和0.31 ± 0.08 mM 2-八醇在一小时内).

结论:

无诱导剂的Pylb促进剂对Bacillus subtilis中的细胞染色体P450表达是有效的.
这种基于Pylb的系统显示出作为可持续生物炼油工艺的生物催化剂的巨大潜力.

关键词:

这种细菌是 Bacillus subtilis.细菌的P450细胞染色体.没有诱导剂的促进剂.

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