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structure of patulin-detoxifying enzyme Y155F/V187K with NADPHstructure of patulin-detoxifying enzyme Y155F/V187K with NADPH
Structural highlights
FunctionPublication Abstract from PubMedPatulin is a fatal mycotoxin that is widely detected in drinking water and fruit-derived products contaminated by diverse filamentous fungi. CgSDR from Candida guilliermondii represents the first NADPH-dependent short-chain dehydrogenase/reductase that catalyzes the reduction of patulin to the nontoxic E-ascladiol. To elucidate the catalytic mechanism of CgSDR, we solved its crystal structure in complex with cofactor and substrate. Structural analyses indicate that patulin is situated in a hydrophobic pocket adjacent to the cofactor, with the hemiacetal ring orienting toward the nicotinamide moiety of NADPH. In addition, we conducted structure-guided engineering to modify substrate-binding residue V187 and obtained variant V187F, V187K and V187W, whose catalytic activity was elevated by 3.9-, 2.2- and 1.7-fold, respectively. The crystal structures of CgSDR variants suggest that introducing additional aromatic stacking or hydrogen-bonding interactions to bind the lactone ring of patulin might account for the observed enhanced activity. These results illustrate the catalytic mechanism of SDR-mediated patulin detoxification for the first time and provide the upgraded variants that exhibit tremendous potentials in industrial applications. Structure-based rational design of a short-chain dehydrogenase/reductase for improving activity toward mycotoxin patulin.,Dai L, Li H, Huang JW, Hu Y, He M, Yang Y, Min J, Guo RT, Chen CC Int J Biol Macromol. 2022 Dec 1;222(Pt A):421-428. doi: , 10.1016/j.ijbiomac.2022.09.121. Epub 2022 Sep 20. PMID:36176222[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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