Substrate specificity study of zearalenone lactonase by analyzing interaction networks of residues near the β6-α6 region

Recently, how could microbial lactonase react to the mycotoxin zearalenone (ZEN) and its derivatives such as α-zearalenol (α-ZOL) is still unclear, resulting in limited applications. In this study, the interaction networks of residues near the β6-α6 region in lactonase from Monosporascus sp. GIB2 (ZENM) were analyzed. As a result, the residue M157 in the β6-α6 region was found significant to the specificity of ZENM, and two mutants including ZENM and ZENM that exhibited higher degradation activity than the wild-type (WT) against α-ZOL was achieved. The molecular dynamics simulation showed that the binding free energy of ZENM and ZENM was -38.68 and -40.84 Kcal/mol for α-ZOL, much lower than the wild-type enzyme (-33.03 Kcal/mol). Moreover, approximately a 54° torsion of the C6' hydroxyl group in α-ZOL was presented in mutants ZENM and ZENM conformation, resulting in a shorter distance between the catalytic pocket and α-ZOL.