The Protein Engineering of Zearalenone Hydrolase Results in a Shift in the PH Optimum of the Relative Activity of the Enzyme

Overview

Journal Toxins (Basel)

Publisher MDPI

Specialty Toxicology

Date 2024 Dec 27

PMID 39728798

Authors

Anna Dotsenko

Igor Sinelnikov

Ivan Zorov

Yury Denisenko

Aleksandra Rozhkova

Larisa Shcherbakova

Affiliations

Soon will be listed here.

Abstract

An acidic shift in the pH profile of zearalenone hydrolase (ZHD), the most effective and well-studied zearalenone-specific lactone hydrolase, is required to extend the range of applications for the enzyme as a decontamination agent in the feed and food production industries. Amino acid substitutions were engineered in the active center of the enzyme to decrease the pKa values of the catalytic residues E126 and H242. The T216K substitution provided a shift in the pH optimum by one unit to the acidic region, accompanied by a notable expansion in the pH profile under acidic conditions. The engineered enzyme demonstrated enhanced activity within the pH range of 3-5 and improved the activity within the pH ranging from 6 to 10. The D31N and D31A substitutions also resulted in a two-unit shift in the pH optimum towards acidic conditions, although this was accompanied by a significant reduction in the enzyme activity. The D31S substitution resulted in a shift in the pH profile towards the alkaline region. The alterations in the enzyme properties observed following the T216K substitution were consistent with the conditions required for the ZHD application as decontamination enzymes at acidic pH values (from 3.0 to 6.0).

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