Biotechnology and Bioprocess Engineering:BBE

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
권호 표지

High Level Production of Supra Molecular Weight Poly(3-Hydroxybutyrate) by Metabolically Engineered Escherichia coli

  • Park, Jong-il (Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical & Biomolecular Engineering and BioProcess Engineering Research Center) ;
  • Lee, Sang-Yup (Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical & Biomolecular Engineering and BioProcess Engineering Research Center, Department of BioSystems and Bioinformatics Research Center, Korea Advanced Institute of Science and Technology)
  • Published : 2004.06.01
Download PDF
⟨ Previous Next ⟩

Abstract

The supra molecular weight poly(〔R〕-3-hydroxybutyrate) (PH B), having a molecular weight greater than 2 million Da, has recently been found to possess improved mechanical properties compared with the normal molecular weight PHB, which has a molecular weight of less than 1 million Da. However, applications for this PHB have been hampered due to the difficulty of its production. Reported here, is the development of a new metabolically engineered Escherichia coli strain and its fermentation for high level production of supra molecular weight PHB. Recombinant E. coli strains, harboring plasm ids of different copy numbers containing the Alcaligenes latus PHB biosynthesis genes, were cultured and the molecular weights of the accumulated PHB were compared. When the recombinant E. coli XL1-Blue, harboring a medium-copy-number pJC2 containing the A. latus PHB biosynthesis genes, was cultivated by fed-batch culture at pH 6.0, supra molecular weight PHB could be produced at up to 89.8 g/L with a productivity of 2.07 g PHB/L-h. The molecular weight of PHB obtained under these conditions was as high as 22 MDa, exceeding by an order of magnitude the molecular weight of PHB typically produced in Ralstonia eutropha or recombinant E. coli.

Keywords

References

  1. Microbiol Rev. v.54 Occurrence, metabolism, metabolic role, and industrial uses of bacterial polyhydroyalkanoates Anderson,A.J.;E.A.Dawes
  2. Biotechnol. Bioeng. v.49 Bacterial polyhydroxyalkanoates Lee,S.Y. https://doi.org/10.1002/(SICI)1097-0290(19960105)49:1<1::AID-BIT1>3.0.CO;2-P
  3. Trends Biotechnol. v.14 Plastic bacteria? Progress and prospects for polyhydroxyalkanoate production in bacteria Lee,S.Y. https://doi.org/10.1016/0167-7799(96)10061-5
  4. Metabolic Engineering Metabolic engineering strategies for the production of polyhydroxyalkanoates, a family of biodegradable polymers Lee,S.Y.;J.Choi;S.Y.Lee(ed.);E.T.Papoutsakis(ed.)
  5. Trends Biotechnol. v.16 Bacterial and other biological systems for polyester production Steinbuchel,A.;B.Fuchtenbusch https://doi.org/10.1016/S0167-7799(98)01194-9
  6. Developments in Crystalline Polymers v.2 Biologically produced PHA polymers and copolymers Holmes,P.A.;D.C.Bassett(ed.)
  7. J. Bacteriol. v.170 Cloning of the Alcaligenes eutrophus poly-β-hydroxybutyrate synthetic pathway and synthesis of PHB in Escherichia coli. Schubert,P.;A.Steinbuchel;H.G.Shlegel https://doi.org/10.1128/jb.170.12.5837-5847.1988
  8. Appl. Environ. Microbiol. v.64 Cloning of the Alcaligenes latus polyhydroxyalkanoate biosynthesis gene and use of these genes for enhanced production of poly(3-hydroxybutyrate) by recombinant Escherichia coli. Choi,J.;S.Y.Lee;K.Han
  9. J. Microbiol. Biotechnol. v.9 Production of poly(3-hydroxybutyrate) [P(3HB)] with high P(3HB) content by recombinant Escherichia coli harboring the Alcaligenes latus P(3HB) biosynthesis genes and the E. coli ftsZ gene Choi,J.;S.Y.Lee
  10. Appl. Environ. Microbiol. v.12 Production of poly(3-hydroxybutyrate) by fed-batch culture of filamentation-suppressed recombinant Escherichia coli Wang,F.;S.Y.Lee
  11. Appl. Microbiol. Biotechnol. v.47 Molecular weight of ploy[(R)-3-hydroybutyric acid] produced in a recombinant Escherichia coli Kusaka,S.;H.Abe;S.Y.Lee;Y.Doi https://doi.org/10.1007/s002530050902
  12. J. Macromol. Sci. Pure Appl. Chem. v.A35 Microbial synthesis and physical properties of ultra-high-molecular-weight poly[(R)-3-hydroxybutyrate] Kusaka,S.;T.Iwata;Y.Doi
  13. Int. J. Biol. Macromol. v.25 Properties and biodegradability of ultra-high-molecular weight poly[(R)-3-hydroxybutyrate] produced by a recombinant Escherichia coli Kusaka,S.;T.Iwata;Y.Doi https://doi.org/10.1016/S0141-8130(99)00019-7
  14. Poly Degrad. Stabil. v.79 Mechanical properties and highly ordered structure of ultra-high-molecular-weight poly [(R)-3-hydroxybutyrate] films: Effect of annealing and two-step drawing Aoyagi,Y.;Y.Doi;T.Iwata https://doi.org/10.1016/S0141-3910(02)00273-2
  15. Int. J. Pharm. v.29 In vitro and in vivo studies on biodegradable polyester microparticles containing sulphamethiazole Brophy,M.R.;P.B.Deasy https://doi.org/10.1016/0378-5173(86)90119-5
  16. Eur. J. Appl. Microbiol. Biotechnol. v.6 A rapid gas chromatographic method for the determination of poly-β-hydroxybutyric acid in microbial biomass Braunegg,G.;B.Sonnleitner;R.M.Lafferty https://doi.org/10.1007/BF00500854
  17. Biotechnol. Bioeng v.62 Efficient and economical recovery of poly(3-hydroxybutyrate) from recombinant Escherichia coli by simple digestion with chemicals Choi,J.;S.Y.Lee https://doi.org/10.1002/(SICI)1097-0290(19990305)62:5<546::AID-BIT6>3.0.CO;2-0
  18. Proc. Natl. Acad. Sci. USA v.92 Enzyme-catalyzed synthesis of poly [(P)-(-)-3-hydroxybutyrate]: Formation of macroscopic granules in vitro Gerngross,T.U.;D.P.Martin https://doi.org/10.1073/pnas.92.14.6279
  19. Appl. Microbiol Biotechnol. v.49 In vitro biosynthesis of poly(3-hydroxybutyric acid) by using purified poly(hydroxyalkanoic acid) synthase of Chromatium vinosum Jossek,R.;R.Reichelt;A.Steinbuchel https://doi.org/10.1007/s002530051166
  20. J. Macromol. Sci. Pure Appl. Chem. v.A35 Molecular weights of poly(3-hydroybutyrate) during biological polymerization in Alcaligenes eutrophus Koizumi,F.;Y.Doi
  21. Nature Biotech. v.15 PHA synthase activity controls the molecular weight and polydispersity of polyhydroxybutyrate in vivo Sim,S.J.;K.D.Snell;S.A.Hogan;J.Stubbe;C.Rha;A.J.Sinskey https://doi.org/10.1038/nbt0197-63
  22. Biotechnol. Bioprocess. Eng. v.7 Pilot scale production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) by fed-batch culture of recombinant Escherichia coli Choi,J.;S.Y.Lee;K.Shin;W.G.Lee;S.J.Park;H.N.Chang;Y.K.Chang https://doi.org/10.1007/BF02933524