9qgh
BV333 aminotransferase from Streptomyces sp. mutant F61C
Structural highlights
Publication Abstract from PubMedA (S)-selective amine transaminase from a Streptomyces strain, Sbv333-ATA, is a biocatalyst showing both high thermostability with a melting temperature of 85 degrees C and broad substrate specificity for the amino acceptor. This enzyme was further characterized both biochemically and structurally. The Sbv333-ATA is stable in the presence of up to 20% (v/v) of the water-miscible cosolvents methanol, ethanol, acetonitrile, and dimethyl sulfoxide, and in biphasic systems with petroleum ether, toluene, and ethyl acetate as an organic phase. The enzyme showed also a good activity toward different amino donors, such as (S)-methylbenzylamine and 2-phenylethylamine, aliphatic mono- and di-amines, like propylamine and cadaverine, and selected amino acids. However, more sterically hindered aromatic amines were not accepted. Based on the knowledge of the three-dimensional structures obtained, a rational approach to site specific mutagenesis was carried out to broaden the substrate specificity of Sbv333-ATA. The mutant W89A showed the highest activity toward bulky amines as substrates, such as the diaromatic compound 1,2-diphenylethylamine. The 3D structures of the holo and inhibitor gabaculine bound forms of native Sbv333-ATA, and holo W89A and F61C mutants were determined at high resolutions of 1.49, 1.24, and 1.31 (both mutants) A, respectively. These structures were important for revealing further details of the active site binding pockets of the Sbv333-ATA and its mechanism. KEY POINTS: * Sbv333-ATA is a highly thermostable transaminase with a broad substrate scope. * Sbv333-ATA remains active in various organic cosolvents and biphasic systems. * Mutant W89A expands substrate range to accept bulky diaromatic amines. Functional and structural insights into a thermostable (S)-selective amine transaminase and its improved substrate scope by protein engineering.,Patti S, De Rose SA, Isupov MN, Magrini Alunno I, Riva S, Ferrandi EE, Littlechild JA, Monti D Appl Microbiol Biotechnol. 2025 Aug 12;109(1):180. doi: , 10.1007/s00253-025-13536-9. PMID:40796981[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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