Methyl-branched poly(hydroxyalkanoate) biosynthesis from 13-methyltetradecanoic acid and mixed isostearic acid isomer substrates

Abstract

Pseudomonas resinovorans, a known medium-chain-length (mcl-) poly(hydroxyalkanoate) (PHA) producer, was grown on 13-methyltetradecanoic acid (13-MTDA) and a mixture of isostearic acid (IA) isomers to produce methyl-branched mcl-PHA polymers. Shake-flask experiments revealed polymer productivities (the percent of the cell mass that is polymer) of 31 +/- 1% (n = 3) and 23 +/- 3% (n = 3) when grown in 13-MTDA and IA, respectively. Monomer content was determined by a combination of gas chromatography/mass spectrometry (GC/MS) of the acid hydrolyzed, silylated methyl esters, and nuclear magnetic resonance spectroscopy. Results showed that the mcl-PHA polymer derived from 13-MTDA was primarily composed of 3-hydroxy-7-methyloctanoic acid and 3-hydroxy-9-methyldecanoic acid (67 and 16 mol% by GC/MS, respectively). In contrast, the mcl-polymers synthesized from the IA isomeric mixture were more complex, containing both even and odd chain-length monomers as well as varying distributions of methyl-branched derivatives. The PHA distributions among the C8, C10, C12, and C14 carbon chain-length monomers included three isomers of C8, five isomers of C10, seven isomers of C12, and nine isomers of C14 each containing one linear-chain derivative and n-6 methyl-branched derivatives where n equals the total number of carbon atoms in each monomer unit (C8-C14).