8JLV

Beneficial flip of substrate orientation enable determine substrate specificity for zearalenone lactone hydrolase

Summary for 8JLV

• Entry DOI: 10.2210/pdb8jlv/pdb
• Descriptor: AB hydrolase-1 domain-containing protein (1 entity in total)
• Functional Keywords: mycotoxin, zearalenone hydrolase, hydrolase
• Biological source: Exophiala aquamarina CBS 119918
• Total number of polymer chains: 6
• Total formula weight: 175885.25

Authors

Xiang, L.,Wang, M.,Zhang, G.,Zhou, J. (deposition date: 2023-06-02, release date: 2023-12-06, Last modification date: 2024-06-19)

Primary citation

Wang, M.,Zhang, F.,Xiang, L.,Li, M.,Lu, Z.,Wu, P.,Sheng, X.,Zhou, J.,Zhang, G.
Enhancing the activity of zearalenone lactone hydrolase toward the more toxic alpha-zearalanol via a single-point mutation.
Appl.Environ.Microbiol., 90:e0181823-e0181823, 2024

PubMed Abstract: Zearalenone (ZEN) and its derivatives are estrogenic mycotoxins known to pose significant health threats to humans and animals. Especially, the derivative α-zearalanol (α-ZAL) is over 10 times more toxic than ZEN. Simultaneous degradation of ZEN and its derivatives, especially α-ZAL, using ZEN lactone hydrolases (ZHDs) is a promising solution to eliminate their potential hazards to food safety. However, most available ZHDs exhibit limited activity toward the more toxic α-ZAL compared to ZEN. Here, we identified a broad-substrate spectrum ZHD, named ZHDAY3, from CBS 119918, which could not only efficiently degrade ZEN but also exhibited 73% relative activity toward α-ZAL. Through rational design, we obtained the ZHDAY3(N153H) mutant, which exhibited the highest specific activity (253.3 ± 4.3 U/mg) reported so far for degrading α-ZAL. Molecular docking, structural comparative analysis, and kinetic analysis collectively suggested that the shorter distance between the side chain of the catalytic residue His242 and the lactone bond of α-ZAL and the increased binding affinity to the substrate were mainly responsible for the improved catalytic activity of ZHDAY3(N153H) mutant. This mechanism was further validated through additional molecular docking of 18 mutants and experimental verification of six mutants.IMPORTANCEThe mycotoxins zearalenone (ZEN) and its derivatives pose a significant threat to food safety. Here, we present a highly promising ZEN lactone hydrolase (ZHD), ZHDAY3, which is capable of efficiently degrading both ZEN and the more toxic derivative α-ZAL. Next, the ZHDAY3(N153H) mutant obtained by single-point mutation exhibited the highest specific activity for degrading α-ZAL reported thus far. We further elucidated the molecular mechanisms underlying the enhanced hydrolytic activity of ZHDAY3(N153H) toward α-ZAL. These findings represent the first investigation on the molecular mechanism of ZHDs against α-ZAL and are expected to provide a significant reference for further rational engineering of ZHDs, which will ultimately contribute to addressing the health risks and food safety issues posed by ZEN-like mycotoxins.

PubMed: 38332488
DOI: 10.1128/aem.01818-23

Experimental method

X-RAY DIFFRACTION (2.99863596858 Å)