脱乙酶的异质表达及其在L-glufosinate制剂中的应用
Yuan-Shan Wang1,2,3, Mei-Hua Gong1,2,3, Jin-Hao Wang1,2,3
1Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, No. 18, Chaowang Road, Hangzhou, 310014, Zhejiang, People's Republic of China.
Bioprocess and biosystems engineering
|September 21, 2023
概括
研究人员开发了一种新的生物催化方法,用于合成L-glufosinate,使用一种新型的脱乙酶NAP-Das2.3.3. 这种在Pichia pastoris中表达的酶有效地产生高纯度的L-glufosinate,为工业生物合成提供了一个有前途的替代品.
科学领域:
- 生物催化剂是一种生物催化剂.
- 酵素工程是什么意思 酵素工程
- 工业生物技术 工业生物技术
背景情况:
- 酸是一种强大的除草剂,具有重要的商业利益.
- 目前的合成方法在效率和立体选择性方面面临挑战.
- 生物催化提供了一种可持续和精确的方法来生产L-glufosinate.
研究的目的:
- 为了异质地表达和表征NAP-Das2.3,一种来自Arenimonas malthae的脱乙酸酶.
- 为了优化NAP-Das2.3的表达和活性,用于L-glufosinate生物合成.
- 评估NAP-Das2.3在转化N-乙-L-糖酸盐到L-糖酸盐中的立体选择效率.
主要方法:
- 在大肠杆菌和牧羊中表达了NAP-Das2.3.
- 在各种发酵条件下 (摇瓶,生物反应器) 评估了酶活性.
- 确定了酶动力学和最佳反应条件 (温度,pH).
主要成果:
- 再组合NAP-Das2.3在大肠杆菌中表现出有限的活性,这是由于包含体的形成.
- 在Pichia pastoris生物反应器发酵中实现了高效的可溶性表达和高活性 (1287.52 U/L).
- 在45°C和pH值8.0下观察到最佳活性,L-glufosinate的基质转化率高 (92.71%) 和反体过量 (>99.9%).
结论:
- 在Pichia pastoris中,NAP-Das2.3的异质表达是有效的L-glufosinate生物合成的可行策略.
- 描述的NAP-Das2.3酶表现出高的立体选择性和活性,使其成为一个有前途的生物催化剂.
- 这项研究为工业规模生产纯L-glufosinate提供了一个潜在的替代方案.
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