罕见的金氏化物化合物K的高效生物合成使用工程Pichia pastoris与表面定Sulfolobus solfataricus的β-Glucosidase
Yizi Luo1,2, Pan Wang1,2, Ruyao Wang1,2
1Engineering Research Center of Western Resource Innovation Medicine Green Manufacturing, Ministry of Education, School of Chemical Engineering, Northwest University, Xi'an 710069, China.
Journal of agricultural and food chemistry
|December 30, 2025
概括
研究人员使用工程酵母改进了罕见的金氏化物化合物K (CK) 的生产. 这种全细胞固定方法提高了酶的稳定性和工业应用的经济可行性.
科学领域:
- 生物技术是生物技术.
- 酵素工程是什么? 酶工程是什么
- 微生物工程 微生物工程
背景情况:
- 苏尔福洛布斯·索尔法塔里克斯β-葡萄糖酶 (SS-bgly) 对于将人参化物转化为K (CK) 化合物至关重要.
- 现有的方法在工业环境中面临酶稳定性和成本效益方面的挑战.
- 需要新的策略来提高SS-bgly的性能,以便在实际CK生产中使用.
研究的目的:
- 开发一种强大的全细胞生物催化剂,以高效稳定地生产化合物K (CK).
- 通过酶固定和应变工程来克服自由酶应用的局限性.
- 优化发酵条件以最大限度地提高CK产量和转化效率.
主要方法:
- 在基因上将SS-bgly定在Pichia pastoris上,使用GPI定GCW61蛋白进行全细胞固定.
- 采用ARTP突变发生因子用于定向进化,以改善酶活性和稳定性.
- 通过协同表达N-乙转移酶MPR1.1,对增强氧化应激抗性的菌株进行工程设计.
- 优化发酵参数以提高β-葡萄糖酶活性和CK生产.
主要成果:
- 与原始菌株相比,工程菌株Cbg61-T37-MPR1的β-葡萄糖酶活性增加了58.9%.
- 实现了93.79%的转化率和6.42 mg/mL的高CK产量.
- 固定全细胞生物催化剂表现出显著的稳定性,在五个周期后保持了63.17%的效率.
结论:
- 开发的酶表面显示,定向进化和反应优化的策略有效地提高了CK的产量.
- 这种方法提高了使用SS-bgly用于工业化合物K合成的实用性和经济可行性.
- 工程酵母菌株为人参化物生物转化提供了一个稳定高效的平台.
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