Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing oxygen-containing organic compound
Patent
1996-07-26
1998-09-22
McKelvey, Terry A.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Preparing oxygen-containing organic compound
435141, 435243, 435375, C12P 762, C12P 752, C12N 100, C12N 502
Patent
active
058112723
ABSTRACT:
A method has been developed for control of molecular weight and molecular weight dispersity during production of polyhydroxyalkanoates in genetically engineered organism by control of the level and time of expression of one or more PHA synthases in the organisms. The method was demonstrated by constructing a synthetic operon for PHA production in E. coli in which the level of PHA synthase activity could be tightly controlled by placement of the synthase behind an inducible promoter. Modulation of the total level of PHA synthase activity in the host cell by varying the concentration of the inducer, isopropyl .beta.-D-thiogalactoside (IPTG), was found to effect the molecular weight of the polymer produced in the cell. Specifically, high concentrations of synthase activity were found to yield polymers of low molecular weight while low concentrations of synthase activity yielded polymers of higher molecular weight. Polymer molecular weight dispersity is also proportional to the amount of synthase activity, with less dispersity in polyhydroxyalkanoate compositions produced in expression systems with an initial burst of synthase activity, and higher levels of molecular weight dispersity in polyhydroxyalkanoate compositions produced in expression systems with the levels of synthase activity varied during synthesis of the polyhydroxyalkanoate.
REFERENCES:
patent: 4477654 (1984-10-01), Holmes et al.
patent: 5245023 (1993-09-01), Peoples et al.
Anderson and Dawes, Occurrence, Metabolism, Metabolic Role, and Industrial Uses of Bacterial Polyhydroxyalkanoates, Microbiological Reviews, 54(4), 450-472 (1990).
Berg, D.E., et al., "The Prokaryotic Transposable Element Tn5," Biotechnology, 1, 417-135 (1983).
Berndt and Schlegel, "Kinetics and Properties of .beta.-Ketothiolase from Clostridium pasteurianum," to Arch. Microbiol., 103, 21-30 (1975).
deSmet, et al., "Characterization of Intracellular Inclusions Formed by Pseudomonas olevorans During Growth on Octane," J. Bacteriol., 154, 870-878 (1983).
Ditta, et al., "Broad Host Range DNA Cloning System for Gram-Negative Bacteria: Construction of a Gene Bank of Rhizobium melioti," Proc. Natl. Acad. Sci. USA, 77, 7347-7351 (1980).
Fukui et al., "Enzymatic Synthesis of Poly-.beta.-Hydroxybutyrate in Zoogloea ramigera," Microbiol. 1976, 110, 149-156.
Fukui, et al., "Purification and Characterization of NADP-linked acctoacetyl-CoA Reductase from Zoogloea ramigera 1-16-M," Biochem. Biophys. Acta, 917, 365-371 (1987).
Gerngross et al., "Overexpression and Purification of the Soluble Polyhydroxyalkanoate Synthase from Alcaligense eutrophus: Evidence for a Required Posttranslational Modification for Catalytic Activity," Biochemistry, 33, 9311-9320 (1994).
Gerngross and Martin, "Enzyme-catalyzed synthesis of vitro,," Proc. Natl. Acad. Sci. USA 92, 6279-6283 (Jul. 1995).
Gilbert, "Molecular Weight Distributions in Free-Radical Polymerizations: Their Cause and Cure" Trends in Polymer Science 3, 222-226 (1995).
Griebel and Merrick, "Metabolism of Poly-.beta.-Hydroxybutyrate: Effect of Mild Alkaline Extraction on Native Poly-Hydroxybutyrate Granules," J. Bacteriol, 108, 782-789 (1971).
Guzman et al., "Tight Regulation, Modulation, and High-Level Expression of Vectors Containing the Arabinose P.sub.BAD Promoter," J. Bacteriol. 177, 4121-4130, 1995.
Hahn et al., "Communication to the Editor: Optimization of Microbial Poly(3-hydroxybutyrate) Recovery Using Dispersions of Sodium Hypochlorite Solution and Chloroform," Biotechnology and Bioengineering, 44, 256-261 (1994).
Haywood et al., "The Importance of PHB-synthase Substrate Specificity in Polyhydroxyalkanoate Synthesis of Alcaligenes eutrophus," FEMS Microbiology Letters, 57, 1-6 (1989).
Huang and Reusch, "Genetic Competence in Escherichia coli Requires Poly-.beta.-Hydroxybutyrate/Calium Polyphosphate Membrane Complexes and Certain Divalent Cations," J. Bacteriol. 177(2), 486-490 (1995).
Huisman, et al., "Metabolism of Poly(3-hydroxyalkanoates) (PHAs) by Pseudomonas oleovorans," J. Biol. Chem. 266(4), 2191-2198 (1991).
Hustede, et al., "Characterization of the Polyhydroxyalkanoate Synthease Gene Locus of Rhodobacter sphaeroides," Biotechnol. Lett. 15(7), 709-714 (1993).
Kawaguchi and Doi, "Kinetics nd Mechanism of Synthesis and Degradation of Poly(3-hydroxybutyrate) in Alcaligenses eutrophus," Macromolecules 25, 2324-2329 (1992).
Kidwell et al., "Regulated Expression of the Alcaligenese eutrophus pha Biosynthesis Genes in Escherichia coli", Applied and Environmental Microbiology 61(4), 1391-1398 (1995).
Kim et al., "Production of Poly-.beta.-Hydroxybutyrate by Fed-Batch Culture of Recombinant Escherichia coli," Biotechnology Letters, 14(9), 811-818 (1992).
Klechner, N., et al., "Genetic Engineering in Vivo Using Translocatable Drug-resistance Elements," J. Mol. Biol., 116, 125-159 (1977).
Koizumi, et al., "Molecular Weight of Poly(3-hyroxybutyrate) During Biological Polymerization in Alcaligenes eutrophus," Chemical Abstracts, 124:50315f (1996).
Law and Slepecky, "Assay of Poly-.beta.-Hydroxybutyric Acid," J. Bacteriol. 82, 33-36 (1961).
Lee, et al., "Construction of Plasmids, Estimation of Plasmid Stability, and Use of Stable Plasmids for the Production of Poly(3-Hydroxybutyric acid) by Recombinant Escherichia coli," Journal of Biotechnology 32, 203-211 (1994).
Lee, S. Y., "Bacterial Polyhydroxyalkanoates," Biotechnology and Bioengineering 49, 1-14, (1996).
Lee, et al., "Production of Poly(.beta.-Hydroxybutyric Acid) by Recombinant Escherichia coli.sup.a ", Annals of the New Your Academy of Sciences, 721, 43-53 (1994).
Liebergesell, et al., "Cloning and Nucleotide Sequences of Genes Relevant for Biosynthesis of Poly(3-Hydroxybutric acid) in Chromatium vinosum strain D," Eur. J. Biochem. 209, 135-150 (1992).
Liebergesell, et al., "Cloning and Molecular Analysis of the Poly(3-Hydroxybutyric Acid) Biosynthetic Genes of Thiocystis violacea," Appl. Microbiol. Biotechnol., 38, 493-501 (1993).
Nawrath et al., "Targeting of the Polyhydroxybutyrate biosynthetic pathway to the Plastids of Arabidopsis thaliana results in high levels of polymer accumulation," Proc. Natl. Acad. Sci USA 91, 12760-12764, 1994.
Nishimura et al., "Purification and Properties of .beta.-Ketothiolase from Zoogloea ramigera," Arch. Microbiol. 1978, 116, 21-27.
Oeding and Schlegel, .beta.-Ketothiolase from Hydrogenomonas eutropha H16 and its Significance in the Regulation of Poly-.beta.-hydroxybutyrate Metabolism, Biochm. J., 134, 239-248 (1973).
Peoples and Sinskey, "Poly-.beta.-hydroxybutyrate Biosynthesis in Alicaligenes eutrophus H16," J. Biol. Chem. 1989, 264, 15293-15297.
Peoples and Sinskey, "Poly-.beta.-Hydroxybutyrate (PHB) Biosynthesis in Alcaligense eutrophus H16," J. Biol. Chem. 1989, 264, 15298-15303.
Peoples, et al., "Biosynthetic Thiolase fromZoogloea ramigera," J. Biol. Chem., 262, 97-102 (1987).
Pieper, et al., "Identification Cloning and Sequence Analysis of the Poly(3-Hydroxyalkanoic Acid) Synthase Gene of the Gram-Positive bacterium Rhodococcus ruber," FEMS Microbiol. Lett., 96, 73-80 (1992).
Ploux, et al., "the NADPH-linked Acetoacetyl-CoA-Reductase from Zoogloea ramigera: Characterization and Mechanistic Studies of the Cloned Enzyme Over-Produced in Escherichia coli," Eur. J. Biochem. 174, 177-182 (1988).
Poirier et al., "Production of Polyhydroxyalkanoates, a Family of Biodegradable Plastics and Elastomers, in Bacteria and Plants," Bio/Technology 13, 142-150 (1995).
Rhie and Dennis, "Role of fadR and atoC(Con) Mutations in Poly(3-Hydroxybutyrate-Co-3-Hydroxyvalerate) Synthesis in Recombinant ph Escherichia coli," Applied and Environmental Microbiology, 1995, 61, 2487-2492.
Saito, et al., "An NADP-Linked Acetoacetyl CoA Reductase from Zoogloea ramigera," Arch. Microbiol., 114, 211-217 (1977).
Schembri, et al., "Cloning and Analysis of the Polyhydroxyalkanoic acid Synthase Gene from an Acinetobacter sp.: Evidence that the Gene is Both Plasmid and Chromosomally Located," FEMS Microbiol. Lett. 118, 145-152 (1994).
Schlegel, et al., "The Isolation of Mutants Not Accumulating Poly-.beta.-Hydroxybutyric Acid," Arch. Microbiol., 71
Hogan Scott A.
Rha ChoKyun
Sim Sang Jun
Sinskey Anthony J.
Snell Kristi D.
Massachusetts Institute of Technology
McKelvey Terry A.
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