Browse > Article
http://dx.doi.org/10.9713/kcer.2017.55.3.363

Inhibition of poly 3-hydroxybutyrate (PHB) synthesis by phaR deletion in Methylobacterium extorquens AM1  

Kim, Yujin (Department of Chemical and Biomolecular Engineering, Sogang University)
Lee, Kwanghyun (Department of Life Sciences, Sogang University)
Kim, Hyeonsoo (Department of Chemical and Biomolecular Engineering, Sogang University)
Cho, Sukhyeong (C1 Gas Refinery R&D Center, Sogang University)
Lee, Jinwon (Department of Chemical and Biomolecular Engineering, Sogang University)
Publication Information
Korean Chemical Engineering Research / v.55, no.3, 2017 , pp. 363-368 More about this Journal
Abstract
Methylotrophy is able to use reduced one-carbon compound, such as methanol and methylamine, as a sole carbon source. Methylobacterium extorquens AM1 is the most extensively studied methylotroph utilizing serine-isocitrate lyase cycle. Because the Poly 3-hydroxybutyrate (PHB) synthesis pathway in M. extorquens AM1 is likely to interlink with EMCP (ethylmalonyl-CoA pathway), glyoxylate, and TCA cycles, regulation of PHB production is needed to produce EMCP-derived acid or TCA acids. To adjust carbon flux to PHB production, PhaR, which seems to have function of regulator of PHB synthesis and acetyl-CoA flux, was knocked out in M. extorquens AM1 by using markerless gene deletion methods. As a result, PHB granules were remarkably reduced in the knockout strain ${\Delta}phaR$ compared to parental strain. Although lag phase was extended for 12h, ${\Delta}phaR$ showed similar cell growth and methanol consumption rate compared to wild type.
Keywords
PhaR; PHB; Methylobacterium extorquens AM1; poly(3-hydroxybutyrate) granule formation;
Citations & Related Records
Times Cited By KSCI : 1  (Citation Analysis)
연도 인용수 순위
1 Salem, A. R. and Quayle, J. R., "Mutants of Pseudomonas AM1 that Require Glycollate or Glyoxylate for Growth on Methanol and Ethanol," Biochem. J., 124, 74(1971).   DOI
2 Chistoserdova, L., Chen, S. W., Lapidus, A. and Lidstrom, M. E., "Methylotrophy in Methylobacterium extorquens AM1 from a Genomic Point of View," J. Bacteriol., 185(10), 2980-2987(2003).   DOI
3 Chistoserdova, L., Metabolism of formaldehyde in M. extorquens AM1: Microbial Growth on C1 Compounds, Springer Netherlands, 16-24(1996).
4 Marx, C. J. and Lidstrom, M. E., "Development of Improved Versatile Broad-host-range Vectors for Use in Methylotrophs and Other Gram-negative Bacteria," Microbiology, 147(8), 2065-2075 (2001).   DOI
5 http://www.integratedgenomics.com/genomereleases.html#list6.
6 Anderson, A. J. and Dawes, E. A., "Occurrence, Metabolism, Metabolic Role, and Industrial Uses of Bacterial Polyhydroxyalkanoates," Microbiol Rev., 54(4), 450-472(1990).
7 Ginige, M. P., Hugenholtz, P., Daims, H., Wagner, M., Keller, J., and Blackall, L. L., "Use of Stable-isotope Probing, Full-cycle rRNA Analysis, and Fluorescence in situ Hybridization-microautoradiography to Study a Methanol-fed Denitrifying Microbial Community," Appl. Environ. Microbial., 70(1), 588-596(2004).   DOI
8 Korotkova, N., Chistoserdova, L. and Lidstrom, M. E., "Poly-${\beta}$- hydroxybutyrate Biosynthesis in the Facultative Methylotroph Methylobacterium extorquens AM1: Identification and Mutation of gap11, gap20, and phaR," J. Bacteriol., 184(22), 6174-6181(2002).   DOI
9 Korotkova, N. and Lidstrom, M. E., "Connection Between Poly-${\beta}$-Hydroxybutyrate Biosynthesis and Growth on $C_1$ and $C_2$ Compounds in the Methylotroph Methylobacterium extorquens AM1," J. Bacteriol., 183(3), 1038-1046(2001).   DOI
10 Hou, C.-T., Methylotrophs: Microbiology, biochemistry, and genetics, CRC pp. 1-53(1984).
11 Faust, U. and Pr Fa, H., "Biomass from Methane and Methanol," Biotechnology. VCH Weinheim 3, 84(1991).
12 Kim, P., Kim, J.-H. and Oh, D.-K., "Improvement in Cell Yield of Methylobacterium sp. by Reducing the Inhibition of Medium Components for Poly-${\beta}$-hydroxybutyrate Production," World J. Microbiol. Biotechnol., 19, 357(2003).   DOI
13 Schrader, J., Schilling, M., Holtmann, D., Sell, D., Villela Filho, M., Marx, A. and Vorholt, J. A., "Methanol-based Industrial Biotechnology: Current Status and Future Perspectives of Methylotrophic Bacteria," Trends Biotechnol., 27, 107(2009).   DOI
14 Van Dien, S. J., Strovas, T. and Lidstrom, M. E., "Quantification of Central Metabolic Fluxes in the Facultative Methylotroph Methylobacterium extorquens AM1 Using $^{13}C$-label Tracing and Mass Spectrometry," Biotechnol. Bioeng., 84, 45-55(2003).   DOI
15 Schafer, A., Tauch, A., Jager, W., Kalinowski, J., Thierbach, G., and Puhler, A., "Small Mobilizable Multi-purpose Cloning Vectors Derived from the Escherichia coli Plasmids pK18 and pK19: Selection of Defined Deletions in the Chromosome of Corynebacterium glutamicum," Gene, 145(1), 69-73(1994).   DOI
16 Lidstrom, M. E., Murrell, J. C. and Dalton, H., (Ed.), The genetics and molecular biology of methanol-utilizing bacteria: Methane and Methanol Utilizers, Springer US, 183-206(1992).
17 Anthony, C., The Biochemistry of Methylotrophs, Academic Press, London, United Kingdom (1982).
18 Senior, P. J. and Windass, J., "The ICI Single Cell Protein Process," J. Biotechnol. Lett., 2(5), 205-210(1980).   DOI
19 Asenjo, J. A. and Suk, J. S., J. Fem Technol., 64, 271-2789(1986).   DOI
20 Chistoserdova, L., "Modularity of Methylotrophy, Revisited," Environ. Microbiol., 13, 2603-2622(2011).   DOI
21 Kim, H. G., Han, G. H., Eom, C. Y. and Kim, S. W., "Isolation and Taxonomic Characterization of a Novel type I Methanotrophic Bacterium," J. Bacteriol., 46(1), 45-50(2008).
22 Degelau, A., Scheper, T., Bailey, J. E. and Guske, C., "Fluorometric Measurement of Poly-${\beta}$ Hydroxybutyrate in Alcaligenes eutrophus by Flow Cytometry and Spectrofluorometry," Appl Microbiol Biotechnol., 42(5), 653-657(1995).   DOI
23 Spiekermann, P., Rehm, B. H., Kalscheuer, R., Baumeister, D., and Steinbüchel, A., "A Sensitive, Viable-colony Staining Method Using Nile Red for Direct Screening of Bacteria that Accumulate Polyhydroxyalkanoic Acids and Other Lipid Storage Compounds," Arch Microbiol., 171(2), 73-80(1999).   DOI
24 Lee, O. K., Hur, D. H., Nguyen, D. T. N. and Lee, E. Y., "Metabolic Engineering of Methanotrophs and Its Application to Production of Chemicals and Biofuels from Methane," Biofuels, Bioprod. Bioref. (2016).
25 Lee, W., "Selection of Medium Components by Plackett-Burman Design for Cell Growth of a Newly Isolated Methylobacterium sp. WJ4," Korean Chem. Eng. Res., 54(6), 812-816(2016).   DOI