title: Rational assignment of key motifs for function guides in silico enzyme identification.
abstract: Biocatalysis has emerged as a powerful alternative to traditional chemistry, especially for asymmetric synthesis. One key requirement during process development is the discovery of a biocatalyst with an appropriate enantiopreference and enantioselectivity, which can be achieved, for instance, by protein engineering or screening of metagenome libraries. We have developed an in silico strategy for a sequence-based prediction of substrate specificity and enantiopreference. First, we used rational protein design to predict key amino acid substitutions that indicate the desired activity. Then, we searched protein databases for proteins already carrying these mutations instead of constructing the corresponding mutants in the laboratory. This methodology exploits the fact that naturally evolved proteins have undergone selection over millions of years, which has resulted in highly optimized catalysts. Using this in silico approach, we have discovered 17 (R)-selective amine transaminases, which catalyzed the synthesis of several (R)-amines with excellent optical purity up to >99% enantiomeric excess.
authors.affiliation: Department of Biotechnology and Enzyme Catalysis, Institute of Biochemistry, Greifswald University, Greifswald, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Höhne
authors.forename: Sebastian
authors.initials: S
authors.lastname: Schätzle
authors.forename: Helge
authors.initials: H
authors.lastname: Jochens
authors.forename: Karen
authors.initials: K
authors.lastname: Robins
authors.forename: Uwe T
authors.initials: UT
authors.lastname: Bornscheuer
mesh_terms: D000465:Algorithms; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D001419:Bacteria / Q000201:enzymology*; D055162:Biocatalysis; D019295:Computational Biology / Q000379:methods*; D030562:Databases, Protein; D018698:Glutamic Acid / Q000737:chemistry / Q000378:metabolism; D007656:Ketoglutaric Acids / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D019289:Pyruvic Acid / Q000737:chemistry / Q000378:metabolism; D016415:Sequence Alignment; D013237:Stereoisomerism; D013329:Structure-Activity Relationship; D013379:Substrate Specificity; D000637:Transaminases / Q000032:analysis* / Q000737:chemistry* / Q000145:classification / Q000378:metabolism
chemical_list: D007656:Ketoglutaric Acids; D018698:Glutamic Acid; D019289:Pyruvic Acid; D000637:Transaminases