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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

Youri M van Nuland, Fons A de Vogel, Gerrit Eggink, Ruud A Weusthuis

Microb Biotechnol. 2017 May;10(3):594-603.

PMID: 28321989

Abstract:

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.

Chemicals Related in the Paper:

Catalog Number Product Name Structure CAS Number Price
AP1731846 Methyl nonanoate Methyl nonanoate 1731-84-6 Price
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