Biochemical characterization and molecular modification of a zearalenone hydrolyzing enzyme Zhd11D from Phialophora attinorum

Zhaoxiang Wang; Feifan Luo; Sijing Jiang; Jonathan Nimal Selvaraj; Yuling Zhou; Guimin Zhang

doi:10.1016/j.enzmictec.2023.110286

Biochemical characterization and molecular modification of a zearalenone hydrolyzing enzyme Zhd11D from Phialophora attinorum

Enzyme Microb Technol. 2023 Oct:170:110286. doi: 10.1016/j.enzmictec.2023.110286. Epub 2023 Jul 10.

Authors

Zhaoxiang Wang¹, Feifan Luo², Sijing Jiang¹, Jonathan Nimal Selvaraj¹, Yuling Zhou³, Guimin Zhang⁴

Affiliations

¹ State Key Laboratory of Biocatalysis and Enzyme Engineering, College of Life Sciences, Hubei University, Wuhan 430062, Hubei, China.
² State Key Laboratory of Biocatalysis and Enzyme Engineering, College of Life Sciences, Hubei University, Wuhan 430062, Hubei, China; Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China.
³ State Key Laboratory of Biocatalysis and Enzyme Engineering, College of Life Sciences, Hubei University, Wuhan 430062, Hubei, China. Electronic address: zyl609@hubu.edu.cn.
⁴ College of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, China. Electronic address: zhangguimin@buct.edu.cn.

PMID: 37499311
DOI: 10.1016/j.enzmictec.2023.110286

Abstract

ZEN lactone hydrolase (ZHD) can hydrolyze zearalenone (ZEN) to less or non-toxic product, providing an environment-friendly way for food or feeds-containing ZENs detoxification. Here, a newly identified ZHD from Phialophora attinorum, annotated as Zhd11D, was characterized to exhibit highest activity against ZEN at pH 8.0 and 35 ℃ with a specific activity of 304.7 U/mg, which was far higher than most of the reported ZHDs. A nonspecific protein engineering method was introduced through fusing a segment of amphiphilic short peptide S1 at the N-terminus of Zhd11D, resulting in both improved activity (1.5-fold) and thermostability (2-fold at 40 ℃). Biochemical analysis demonstrated that self-aggregation caused by intermolecular interactions between S1 contributed to the improvement of the enzymatic properties of Zhd11D. Additionally, S1-Zhd11D showed a higher hydrolysis rate of ZEN than Zhd11D in peanut oil.

Keywords: Amphiphilic peptide; Enzymatic characterization; ZEN hydrolysis; ZEN-hydrolyzing enzyme.

MeSH terms

Hydrolases / metabolism
Lactones
Phialophora / metabolism
Zearalenone* / chemistry
Zearalenone* / metabolism

Substances

Zearalenone
Hydrolases
Lactones

Supplementary concepts

Phialophora attae