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Bioprospecting Reveals Class III omega-Transaminases Converting Bulky Ketones and Environmentally Relevant Polyamines

AuthorsCoscolín, Cristina; Katzke, Nadine; García-Moyano, Antonio; Navarro-Fernández, José; Almendral, David; Martínez-Martínez, Mónica CSIC ORCID; Bollinger, Alexander; Bargiela, Rafael CSIC ORCID CVN; Gertler, Christoph; Chernikova, Tatyana N.; Rojo, David; Barbas, Coral; Tran, Hai; Golyshina, Olga V.; Koch, Rainhard; Yakimov, Michail M.; Bjerga, Gro E. K.; Golyshin, Peter N.; Jaeger, Karl-Erich; Ferrer, Manuel CSIC ORCID; INMARE Consortium
KeywordsAmine transaminases
Chiral aminemeta
Issue Date2019
PublisherAmerican Society for Microbiology
CitationApplied and Environmental Microbiology 85: e02404- e02418 (2019)
AbstractAmination of bulky ketones, particularly in (R)-configuration, is an attractive chemical conversion, however known ω-transaminases (ω-TAs) show insufficient performances. By applying two screening methods we discovered 10 amine transaminases from the class III ω-TAs family, 38% to 76% identical to homologues. We present examples of such enzymes preferring bulky ketones over keto acids and aldehydes with stringent (S)-selectivity. We also report representatives from the class III ω-TAs capable of converting (R) and (S) amines and bulky ketones, and one converting amines with longer alkyl substituents. The preference for bulky ketones was associated with the presence of a hairpin region proximal to the conserved Arg414 and residues conforming and close to it. The outward orientation of Arg414 additionally favored the conversion of (R)-amines. This configuration was also found to favor the utilization of putrescine as amine donor, so that class III ω-TAs with Arg414 in outward orientation may participate in vivo in the catabolism of putrescine. The positioning of the conserved Ser231 contributes also to the preference of amines with longer alkyl substituents. Optimal temperatures for activity ranged from 45 to 65°C, and a few enzymes retained ≥ 50% of their activity in water-soluble solvents (up to 50% (v/v)). Hence, our results will pave the way to design, in the future, new class III ω-TAs converting bulky ketones and (R)-amines for production of high value products, and screen those converting putrescine.
Publisher version (URL)
Identifiersdoi: 10.1128/AEM.02404-18
e-issn:: 1098-5336
issn:: 0099-2240
Appears in Collections:(ICP) Artículos

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