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

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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).

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References

  • Anzai Y, Kim H, Park J-Y, Wakabayashi H, Oyaizu H (2000) Phylogenetic affiliation of the pseudomonads based on 16s rRNA sequence. Int J Syst Evol Microbiol 50:1563–1589

    Article  CAS  PubMed  Google Scholar 

  • Ashby RD, Foglia TA (1998) Poly(hydroxyalkanoate) biosynthesis from triglyceride substrates. Appl Microbiol Biotechnol 49:431–437

    Article  CAS  Google Scholar 

  • Ashby RD, Solaiman DKY, Foglia TA (2002) Poly(ethylene glycol)-mediated molar mass control of short-chain- and medium-chain-length poly(hydroxyalkanoates) from Pseudomonas oleovorans. Appl Microbiol Biotechnol 60:154–159

    Article  CAS  PubMed  Google Scholar 

  • Ashby RD, Solaiman DKY, Foglia TA (2004) Bacterial poly(hydroxyalkanoate) polymer production from the biodiesel co-product stream. J Polym Environ 12:105–112

    Article  CAS  Google Scholar 

  • Ashby RD, Solaiman DKY, Foglia TA (2005) Synthesis of short-/medium-chain-length poly(hydroxyalkanoate) blends by mixed culture fermentation of glycerol. Biomacromolecules 6:2106–2112

    Article  CAS  PubMed  Google Scholar 

  • Brandl H, Gross RA, Lenz RW, Fuller RC (1988) Pseudomonas oleovorans as a source of poly(β-hydroxyalkanoates) for potential applications as biodegradable polyesters. Appl Environ Microbiol 54:1977–1982

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Casini E, de Rijk TC, de Waard P, Eggink G (1997) Synthesis of poly(hydroxyalkanoate) from hydrolyzed linseed oil. J Environ Polym Degrad 5:153–158

    CAS  Google Scholar 

  • Choi MH, Yoon SC (1994) Polyester biosynthesis characteristics of Pseudomonas citronellolis grown on various carbon sources, including 3-methyl-branched substrates. Appl Environ Microbiol 60:3245–3254

    CAS  PubMed  PubMed Central  Google Scholar 

  • Christie WW (1982) Lipid analysis: isolation, separation, identification, and structural analysis of lipids. Pergamon, Oxford New York

    Google Scholar 

  • de Smet M-J, Eggink G, Witholt B, Kingma J, Wynberg H (1983) Characterization of intracellular inclusions formed by Pseudomonas oleovorans during growth on octane. J Bacteriol 154:870–878

    Article  PubMed  PubMed Central  Google Scholar 

  • de Waard P, van der Wal H, Huijberts GNM, Eggink G (1993) Heteronuclear NMR analysis of unsaturated fatty acids in poly(3-hydroxyalkanoates). J Biol Chem 268:315–319

    PubMed  Google Scholar 

  • Doi Y, Kunioka M, Nakamura Y, Soga K (1986) Nuclear magnetic resonance studies on poly(β-hydroxybutyrate) and a copolyester of β-hydroxybutyrate and β-hydroxyvalerate isolated from Alcaligenes eutrophus H16. Macromolecules 19:2860–2864

    Article  CAS  Google Scholar 

  • Eggink G, van der Wal H, Huijberts GNM, de Waard P (1993) Oleic acid as a substrate for poly-3-hydroxybutyrate formation in Alcaligenes eutrophus and Pseudomonas putida. Ind Crops Products 1:157–163

    Article  Google Scholar 

  • Fall RR, Brown JL, Schaeffer TL (1979) Enzyme recruitment allows the biodegradation of recalcitrant branched hydrocarbons by Pseudomonas citronellolis. Appl Environ Microbiol 38:715–722

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Foglia TA, Vail PD (1993) An efficient, large-scale synthesis of triisopentadecanoin. Org Prep and Proc Int 25:209–213

    Article  CAS  Google Scholar 

  • Fritzsche K, Lenz RW, Fuller RC (1990a) Production of unsaturated polyesters by Pseudomonas oleovorans. Int J Biol Macromol 12:85–91

    Article  CAS  PubMed  Google Scholar 

  • Fritzsche K, Lenz RW, Fuller RC (1990b) Bacterial polyesters containing branched poly(β-hydroxyalkanoate) units. Int J Biol Macromol 12:92–101

    Article  CAS  PubMed  Google Scholar 

  • Goddard TD, Kneller DG (2008) SPARKY 3. University of California, San Francisco http://www.cgl.ucsf.edu/home/sparky

    Google Scholar 

  • Gross RA, DeMello C, Lenz RW, Brandl H, Fuller RC (1989) Biosynthesis and characterization of poly(β-hydroxyalkanoates) produced by Pseudomonas oleovorans. Macromolecules 22:1106–1115

    Article  CAS  Google Scholar 

  • Haywood GW, Anderson AJ, Ewing DF, Dawes EA (1990) Accumulation of a polyhydroxyalkanoate containing primarily 3-hydroxydecanoate from simple carbohydrate substrates by Pseudomonas sp. strain NCIMB 40135. Appl Environ Microbiol 56:3354–3359

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hazer B, Lenz RW, Fuller RC (1994) Biosynthesis of methyl-branched poly(β-hydroxyalkanoate)s by Pseudomonas oleovorans. Macromolecules 27:45–49

    Article  CAS  Google Scholar 

  • Huijberts GNM, Eggink G, de Waard P, Huisman GW, Witholt B (1992) Pseudomonas putida KT2442 cultivated on glucose accumulates poly(3-hydroxyalkanoates) consisting of saturated and unsaturated monomers. Appl Environ Microbiol 58:536–544

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lageveen RG, Huisman GW, Preusting H, Ketelaar P, Eggink G, Witholt B (1988) Formation of polyesters by Pseudomonas oleovorans: effect of substrates on formation and composition of poly(R)-3-hydroxyalkanoates and poly(R)-3-hydroxyalkenoates. Appl Environ Microbiol 54:2924–2932

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lau EP, Gibson KM, Fall RR (1980) Alternate microbial strategies for the metabolism of a 3-methyl branched alkanoic acid. Curr Microbiol 4:163–167

    Article  CAS  Google Scholar 

  • Lee EY, Choi CY (1995) Gas chromatography-mass spectrometric analysis and its application to a screening procedure for novel bacterial polyhydroxyalkanoic acids containing long chain saturated and unsaturated monomers. J Ferm Bioeng 80:408–414

    Article  CAS  Google Scholar 

  • Ngo HL, Nuñez A, Lin W, Foglia TA (2007) Zeolite-catalyzed isomerization of oleic acid to branched-chain isomers. Eur J Lipid Sci Technol 108:214–224

    Article  Google Scholar 

  • Pirnik MP, McKenna EJ (1977) Microbial oxidation of methyl branched alkanes. Crit Rev Microbiol 5:413–422

    Article  CAS  Google Scholar 

  • Satoh H, Mino T, Matsuo T (1992) Uptake of organic substrates and accumulation of polyhydroxyalkanoates linked with glycolysis of intracellular carbohydrates under anaerobic conditions in the biological excess phosphate removal process. Wat Sci Technol 26:933–942

    Article  CAS  Google Scholar 

  • Shimamura E, Kasuya K, Kobayashi G, Shiotani T, Shima Y, Doi Y (1994) Physical properties and biodegradability of microbial poly(3-hydroxybutyrate-co-3-hydroxyhexanoate). Macromolecules 27:878–880

    Article  CAS  Google Scholar 

  • Solaiman DKY, Ashby RD, Foglia TA (2002) Physiological characterization and genetic engineering of Pseudomonas corrugata for medium-chain-length polyhydroxyalkanoates synthesis from triacylglycerols. Curr Microbiol 44:189–195

    Article  CAS  PubMed  Google Scholar 

  • Solaiman DKY, Ashby RD, Hotchkiss AT Jr, Foglia TA (2006) Biosynthesis of medium-chain-length poly(hydroxyalkanoates) from soy molasses. Biotechnol Lett 28:157–162

    Article  CAS  PubMed  Google Scholar 

  • Steinbuchel A, Valentin HE (1995) Diversity of bacterial polyhydroxyalkanoic acids. FEMS Microbiol Lett 128:219–228

    Article  Google Scholar 

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Acknowledgment

The authors thank Marshall Reed for his technical assistance throughout the study.

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Correspondence to Richard D. Ashby.

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Ashby, R.D., Ngo, H.L., Solaiman, D.K.Y. et al. Methyl-branched poly(hydroxyalkanoate) biosynthesis from 13-methyltetradecanoic acid and mixed isostearic acid isomer substrates. Appl Microbiol Biotechnol 85, 359–370 (2009). https://doi.org/10.1007/s00253-009-2134-1

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  • DOI: https://doi.org/10.1007/s00253-009-2134-1

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