8CLQ

Zearalenone lactonase of Streptomyces coelicoflavus mutant H286Y in complex with hydrolyzed zearalenone

Summary for 8CLQ

• Entry DOI: 10.2210/pdb8clq/pdb
• Descriptor: Hydrolase, 2,4-dihydroxy-6-[(1E,10S)-10-hydroxy-6-oxoundec-1-en-1-yl]benzoic acid (3 entities in total)
• Functional Keywords: zearalenone, mycotoxin, lactonase, carboxylesterase, esterase, hydrolase, biodegradation, streptomyces coelicoflavus
• Biological source: Streptomyces coelicoflavus
• Total number of polymer chains: 4
• Total formula weight: 140802.21

Authors

Puehringer, D.,Mlynek, G. (deposition date: 2023-02-17, release date: 2024-02-28, Last modification date: 2024-03-20)

Primary citation

Fruhauf, S.,Puhringer, D.,Thamhesl, M.,Fajtl, P.,Kunz-Vekiru, E.,Hobartner-Gussl, A.,Schatzmayr, G.,Adam, G.,Damborsky, J.,Djinovic-Carugo, K.,Prokop, Z.,Moll, W.D.
Bacterial Lactonases ZenA with Noncanonical Structural Features Hydrolyze the Mycotoxin Zearalenone.
Acs Catalysis, 14:3392-3410, 2024

PubMed Abstract: Zearalenone (ZEN) is a mycoestrogenic polyketide produced by and other phytopathogenic members of the genus . Contamination of cereals with ZEN is frequent, and hydrolytic detoxification with fungal lactonases has been explored. Here, we report the isolation of a bacterial strain, PFA D8-1, with ZEN hydrolyzing activity, cloning of the gene encoding α/β hydrolase ZenA encoded on the linear megaplasmid pSFRL1, and biochemical characterization of nine homologues. Furthermore, we report site-directed mutagenesis as well as structural analysis of the dimeric ZenA of and the more thermostable, tetrameric ZenA of with and without bound ligands. The X-ray crystal structures not only revealed canonical features of α/β hydrolases with a cap domain including a Ser-His-Asp catalytic triad but also unusual features including an uncommon oxyanion hole motif and a peripheral, short antiparallel β-sheet involved in tetramer interactions. Presteady-state kinetic analyses for ZenA and ZenA identified balanced rate-limiting steps of the reaction cycle, which can change depending on temperature. Some new bacterial ZEN lactonases have lower and higher than the known fungal ZEN lactonases and may lend themselves to enzyme technology development for the degradation of ZEN in feed or food.

PubMed: 38449531
DOI: 10.1021/acscatal.4c00271

Experimental method

X-RAY DIFFRACTION (1.53 Å)