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Issue 40, 2018, Issue in Progress
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Metagenomic discovery of a novel transaminase for valorization of monoaromatic compounds

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Abstract

The profitability of next-generation biorefineries is acutely contingent on the discovery and utilization of biocatalysts that can valorize lignin. To this end, the metabolic catalogues of diverse microbiota have been mined previously using functional metagenomics in order to identify biocatalysts that can selectively degrade lignin into monoaromatic compounds. Herein, we have further improved the valorization factor of biorefining by deploying functional metagenomics toward the identification of a novel transaminase that can selectively functionalize lignin-derived monoaromatics to produce value-added feedstocks for pharmaceutical synthesis. We implemented a high-throughput colorimetric assay using o-xylylenediamine as the amino donor and successfully identified a transaminase that utilizes the canonical cofactor, pyridoxal 5′-phosphate, to aminate as many as 14 monoaromatic aldehydes and ketones. We subsequently identified the optimal conditions for enzyme activity towards the most favoured amino acceptor, benzaldehyde, including temperature, pH and choice of co-solvent. We also evaluated the specificity of the enzyme towards a variety of amino donors, as well as the optimal concentration of the most favoured amino donor. Significantly, the novel enzyme is markedly smaller than typical transaminases, and it is stably expressed in E. coli without any modifications to its amino acid sequence. Finally, we developed and implemented a computational methodology to assess the activity of the novel transaminase. The methodology is generalizable for assessing any transaminase and facilitates in silico screening of enzyme–substrate combinations in order to develop efficient biocatalytic routes to value-added amines. The computational pipeline is an ideal complement to metagenomics and opens new possibilities for biocatalyst discovery.

Graphical abstract: Metagenomic discovery of a novel transaminase for valorization of monoaromatic compounds

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Supplementary files

Article information


Submitted
30 Mar 2018
Accepted
12 Jun 2018
First published
20 Jun 2018

This article is Open Access

RSC Adv., 2018,8, 22490-22497
Article type
Paper

Metagenomic discovery of a novel transaminase for valorization of monoaromatic compounds

Sandip V. Pawar, S. J. Hallam and V. G. Yadav, RSC Adv., 2018, 8, 22490
DOI: 10.1039/C8RA02764A

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