Identification,heterologous expression and characterization of a new unspecific peroxygenase from Marasmius fiardii PR-910  

作　　者：Xin Fu Kexin Lin Xiaodong Zhang Zhiyong Guo Lixin Kang Aitao Li 

机构地区：[1]State 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

出　　处：《Bioresources and Bioprocessing》2024年第1期477-488,共12页生物资源与生物加工(英文)

基　　金：funded by the National Key Research and Development Program of China(2021YFC2102700);was Supported by the Key Science and Technology Innovation Project of Hubei Province(2021BAD001);as well as the Research Program of State Key Laboratory of Biocatalysis and Enzyme Engineering。

摘　　要：Unspecific peroxygenases(UPOs)are glycosylated enzymes that provide an efficient method for oxyfunctionalizing a variety of substrates using only hydrogen peroxide(H2O2)as the oxygen donor.However,their poor heterologous expression has hindered their practical application.Here,a novel UPO from Marasmius fiardii PR910(MfiUPO)was identified and heterologously expressed in Pichia pastoris.By employing a two-copy expression cassette,the protein titer reached 1.18 g L−1 in a 5 L bioreactor,marking the highest record.The glycoprotein rMfiUPO exhib-ited a smeared band in the 40 to 55 kDa range and demonstrated hydroxylation,epoxidation and alcohol oxidation.Moreover,the peroxidative activity was enhanced by 150%after exposure to 50%(v/v)acetone for 40 h.A semipreparative production of 4-OH-β-ionone on a 100 mL scale resulted in a 54.2%isolated yield with 95%purity.With its high expression level,rMfiUPO is a promising candidate as an excellent parental template for enhancing desirable traits such as increased stability and selectivity through directed evolution,thereby meeting the necessary criteria for practical application.

关 键 词：Unspecific peroxygenase Marasmius fiardii Heterologous expression CHARACTERIZATION Semi-preparative 

分 类 号：O62[理学—有机化学]