Metabolic production of a novel polymer feedstock, 3-carboxy muconate,from vanillin |
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Authors: | Aaron Gosling S Jane Fowler Michael S O’Shea Melissa Straffon Geoff Dumsday Michael Zachariou |
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Institution: | (1) Faculty of Life and Social Sciences, Swinburne University of Technology, Hawthorn, VIC, Australia;(2) Department of Biological Sciences, University of Calgary, Calgary, Canada;(3) CSIRO Materials Science and Engineering, Bayview Avenue, Clayton, VIC, Australia;(4) Future Manufacturing Flagship, CSIRO, Bayview Avenue, Clayton, VIC, Australia;(5) BioMarin Pharmaceutical Inc., Novato, CA, USA |
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Abstract: | Vanillin can be produced on a commercial scale by depolymerising renewable lignin. One product of microbial metabolism of
vanillin by common soil microbes, such as Acinetobacter baylyi, is a tricarboxylic acid with a butadiene backbone known as 3-carboxy muconate (3CM). Three enzymes, 4-hydroxy benzaldehyde
dehydrogenase, vanillate monooxygenase and protocatechuate 3,4-dioxygenase, catalyse the biotransformation of vanillin to
3CM. These three enzymes were metabolically engineered into an Escherichia coli host, giving a biocatalyst that converted vanillin into 3CM. The biocatalyst was found to give 100% yield of 3CM from 1 mM
of vanillin after 39 h. The rate-limiting reaction was identified as the conversion of vanillate to 3,4-dihydroxybenzoate
catalysed by vanillate monooxygenase. Low expression of the reductase subunit of this enzyme was identified as contributing
to the reduced rate of this reaction. Proof of principle of a novel application for 3CM was demonstrated when it was converted
into a trimethyl ester derivative and copolymerised with styrene. |
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