来自Marasmius fiardii的新型非特异性过氧酶的鉴定,异质表达和表征PR-91010
Xin Fu1, Kexin Lin1, Xiaodong Zhang1
1State Key Laboratory of Biocatalysis and Enzyme Engineering, Hubei Key Laboratory of Industrial Biotechnology, School of Life Sciences, Hubei University, #368 Youyi Road, Wuhan, 430062, People's Republic of China.
Bioresources and bioprocessing
|April 22, 2024
概括
研究人员从Marasmius fiardii PR910 (MfiUPO) 开发出了一种新的非特异性过氧酶 (UPO),在Pichia pastoris中达到创纪录的高表达水平. 这一突破增强了UPO在生物催化剂和酶工程中的应用.
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
- 蛋白质表达方式
背景情况:
- 非特异性过氧酶 (UPO) 是一种使用过氧化 (H2O2) 进行基质氧功能化的糖化酶.
- 由于UPO的异质表达不佳,限制了它们的工业和实际应用.
研究的目的:
- 识别和异质地表达一个来自Marasmius fiardii PR910 (MfiUPO) 的新型UPO.
- 为了实现MfiUPO的高水平蛋白质生产,用于潜在的生物催化剂应用.
主要方法:
- 识别和克隆MfiUPO基因.
- 在Pichia pastoris中使用双副本表达盒表达MfiUPO的异构表达.
- 生物反应器培养和表达酶的特征.
主要成果:
- 在5L生物反应器中为MfiUPO实现了1.18g L-1的记录蛋白位.
- 再组合MfiUPO (rMfiUPO) 显示了氧化,环氧化和酒精氧化活动.
- 在乙处理后,酶活性增强了150%,并且得到了54.2%的4-OH-β-ionone产量.
结论:
- MfiUPO表现出高表达水平和多功能催化活性,使其成为一个有前途的生物催化剂.
- rMfiUPO的高表达水平为酶工程提供了坚实的基础,以提高稳定性和选择性.
- 这项工作克服了以前的表达限制,为实际的UPO应用铺平了道路.
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