The identification and use of robust transaminases from a domestic drain metagenome
2019
Leipold, Leona | Dobrijevic, Dragana | Jeffries, Jack W. E. | Bawn, Maria | Moody, Thomas S. | Ward, John M. | Hailes, Helen C.
Transaminases remain one of the most promising biocatalysts for use in chiral amine synthesis, however their industrial implementation has been hampered by their general instability towards, for example, high amine donor concentrations and organic solvent content. Herein we describe the identification, cloning and screening of 29 novel transaminases from a household drain metagenome. The most promising enzymes were fully characterised and the effects of pH, temperature, amine donor concentration and co-solvent determined. Several enzymes demonstrated good substrate tolerance as well as an unprecedented robustness for a wild-type transaminase. One enzyme in particular readily accepted IPA as an amine donor giving the same conversion with 2–50 equivalents, as well as being tolerant to a number of co-solvents, and operational in up to 50% DMSO – a characteristic as yet unobserved in a wild-type transaminase. This work highlights the value of using metagenomics for biocatalyst discovery from niche environments, and here has led to the identification of one of the most robust native transaminases described to date, with respect to IPA and DMSO tolerance.
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