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• W3205056360 abstract "Abstract Hydrogen peroxide is involved in a variety of enzyme catalysis as an oxidant or toxic by‐product. Thereby, attenuation of the H 2 O 2 ‐driven oxidative stress is one of the key issues for preparative biocatalysis. Here, a rational approach to improve the robustness of enzymes, in particular, Baeyer‐Villiger monooxygenases (BVMOs) against H 2 O 2 was investigated. The enzyme access tunnels, which may serve as exit paths for H 2 O 2 from the active site to the bulk, were predicted by using the CAVER and/or protein energy landscape exploration (PELE) software for the phenylacetone monooxygenase variant (PAMO_C65D) from Thermobifida fusca and the BVMO from Pseudomonas putida KT2440. The amino acid residues, which are susceptible to oxidation by H 2 O 2 (e. g., methionine and tyrosine) and located in vicinity of the predicted H 2 O 2 migration paths, were substituted with less reactive or inert amino acids (e. g., leucine and isoleucine). This led to design of the H 2 O 2 ‐resistant enzyme variants, which became robust biocatalysts for synthetic applications. For instance, the H 2 O 2 ‐resistant P. putida BVMO reached turnover numbers of 4,100 for the BV oxygenation of 4‐decanone, which is 2.8‐fold greater than the parent enzyme. Moreover, the H 2 O 2 ‐resistant P. putida BVMO allowed 2‐fold enhancement in titer of 9‐(nonanoyloxy)nonanoic acid (8) formation in a cascade fatty acid biotransformation. Therefore, it was assumed that the CAVER/PELE‐based H 2 O 2 migration path engineering represents an efficient rational design approach to improve not only oxidative stability but also biotransformation performance of the H 2 O 2 ‐forming or utilizing enzymes (e. g., BVMOs, oxidases, and peroxidases). magnified image" @default.
• W3205056360 created "2021-10-25" @default.
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• W3205056360 date "2021-11-05" @default.
• W3205056360 modified "2023-10-04" @default.
• W3205056360 title "Enzyme Access Tunnel Engineering in Baeyer‐Villiger Monooxygenases to Improve Oxidative Stability and Biocatalyst Performance" @default.
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• W3205056360 doi "https://doi.org/10.1002/adsc.202101044" @default.
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