Expansion of the ω‐oxidation system AlkBGTL of Pseudomonas putida GPo1 with AlkJ and AlkH results in exclusive mono‐esterified dicarboxylic acid production in E. coli
نویسندگان
چکیده
The AlkBGTL proteins coded on the alk operon from Pseudomonas putida GPo1 can selectively ω-oxidize ethyl esters of C6 to C10 fatty acids in whole-cell conversions with Escherichia coli. The major product in these conversions is the ω-alcohol. However, AlkB also has the capacity to overoxidize the substrate to the ω-aldehyde and ω-acid. In this study, we show that alcohol dehydrogenase AlkJ and aldehyde dehydrogenase AlkH are able to oxidize ω-alcohols and ω-aldehydes of esterified fatty acids respectively. Resting E. coli expressing AlkBGTHJL enabled exclusive mono-ethyl azelate production from ethyl nonanoate, with an initial specific activity of 61 U gcdw-1 . Within 2 h, this strain produced 3.53 mM mono-ethyl azelate, with a yield of 0.68 mol mol-1 . This strain also produced mono-ethyl dicarboxylic acids from ethyl esters of C6 to C10 fatty acids and mono-methyl azelate from methyl nonanoate. Adding ethyl nonanoate dissolved in carrier solvent bis-(2-ethylhexyl) phthalate enabled an increase in product titres to 15.55 mM in two-liquid phase conversions. These findings indicate that E. coli expressing AlkBGTHJL is an effective producer of mono-esterified dicarboxylic acids from fatty acid esters.
منابع مشابه
Combination of ester biosynthesis and ω-oxidation for production of mono-ethyl dicarboxylic acids and di-ethyl esters in a whole-cell biocatalytic setup with Escherichia coli
BACKGROUND Medium chain length (C6-C12) α,ω-dicarboxylic acids (DCAs) and corresponding esters are important building blocks for the polymer industry. For DCAs of 12 carbon atoms and longer, a sustainable process based on monooxygenase catalyzed ω-oxidation of fatty-acids has been realized. For medium-chain DCAs with a shorter chain length however, such a process has not been developed yet, sin...
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