Discovery of enzymes for toluene synthesis from anoxic microbial communities

Beller, Harry R.; Rodrigues, Andria V.; Zargar, Kamrun; Wu, Yu-Wei; Saini, Avneesh K.; Saville, Renee M.; Pereira, Jose H.; Adams, Paul D.; Tringe, Susannah G.; Petzold, Christopher J.; Keasling, Jay D.

doi:10.1038/s41589-018-0017-4

Discovery of enzymes for toluene synthesis from anoxic microbial communities

Journal Article · Mon Mar 19 00:00:00 EDT 2018 · Nature Chemical Biology

DOI:https://doi.org/10.1038/s41589-018-0017-4· OSTI ID:1530356

^[1]; Rodrigues, Andria V. ^[2]; Zargar, Kamrun ^[2]; ^[3]; Saini, Avneesh K. ^[2]; Saville, Renee M. ^[2]; Pereira, Jose H. ^[1]; Adams, Paul D. ^[4]; ^[5]; ^[2]; ^[6]

Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States)
Taipei Medical Univ. (Taiwan)
Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Univ. of California, Berkeley, CA (United States)
USDOE Joint Genome Institute (JGI), Walnut Creek, CA (United States)
Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Univ. of California, Berkeley, CA (United States); Technical Univ. of Denmark, Lyngby (Denmark)

Microbial toluene biosynthesis was reported in anoxic lake sediments more than three decades ago, but the enzyme catalyzing this biochemically challenging reaction has never been identified. Here we report the toluene-producing enzyme PhdB, a glycyl radical enzyme of bacterial origin that catalyzes phenylacetate decarboxylation, and its cognate activating enzyme PhdA, a radical S-adenosylmethionine enzyme, discovered in two distinct anoxic microbial communities that produce toluene. The unconventional process of enzyme discovery from a complex microbial community (>300,000 genes), rather than from a microbial isolate, involved metagenomics- and metaproteomics-enabled biochemistry, as well as in vitro confirmation of activity with recombinant enzymes. This work expands the known catalytic range of glycyl radical enzymes (only seven reaction types had been characterized previously) and aromatic-hydrocarbon-producing enzymes, and will enable first-time biochemical synthesis of an aromatic fuel hydrocarbon from renewable resources, such as lignocellulosic biomass, rather than from petroleum.

View Accepted Manuscript (DOE)

Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 1530356

Journal Information:: Nature Chemical Biology, Journal Name: Nature Chemical Biology Journal Issue: 5 Vol. 14; ISSN 1552-4450

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: English

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