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Abstract:

A recombinant E. coli strain (K24K) was constructed and evaluated For poly(3-hydroxybutyrate) (PHB) production from whey and corn steep liquor as main carbon and nitrogen sources. This strain bears the pha biosynthetic genes from Azotobacter sp. strain FA8 expressed from a T5 promoter under the control of the lactose operator. K24K does not produce the lactose repressor, ensuring constitutive expression of genes involved in lactose transport and utilization. PHB was efficiently produced by the recombinant strain grown aerobically in fed-batch cultures in a laboratory scale bioreactor on a semisynthetic medium supplemented with the agroindustrial by-products. After 24 h, cells accumulated PHB to 72.9% of their cell dry weight, reaching a volumetric productivity of 2.13 g PHB per liter per hour. Physical analysis of PHB recovered from the recombinants showed that its molecular weight was similar to that of PHB produced by Azotobacter sp. strain FA8 and higher than that of the polymer from Cupriavidus necator and that its glass transition temperature was approximately 20°C higher than those of PHBs from the natural producer strains. Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Registro:

Documento: Artículo
Título:New recombinant Escherichia coli strain tailored for the production of poly(3-hydroxybutyrate) from agroindustrial by-products
Autor:Nikel, P.I.; De Almeida, A.; Melillo, E.C.; Galvagno, M.A.; Pettinari, M.J.
Filiación:Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina
Instituto de Investigaciones Biotecnológicas, Universidad Nacional de General San Martín, Buenos Aires, Argentina
Departamento de Ingeniería Química, Facultad de Ingeniería, Universidad de Buenos Aires, Buenos Aires, Argentina
Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Ciudad Universitaria-Pabellón 2, 1428 Buenos Aires, Argentina
Palabras clave:Bioreactors; Carbon; Cells; Genes; Glass transition; Molecular weight; Nitrogen; Polymers; Volumetric analysis; Azotobacter; Cupriavidus necator; Lactose operator; Liquor; Poly(3-hydroxybutyrate) (PHB); Escherichia coli; carbon; lactose; lactose repressor; nitrogen; poly(3 hydroxybutyric acid); polymer; unclassified drug; bacterium; ester; gene expression; genetic analysis; plant extract; agriculture; alcoholic beverage; article; Azotobacter; bacterial gene; bacterial strain; bioreactor; corn; Escherichia coli; Escherichia coli k24k; evaluation; fed batch culture; gene expression; glass transition temperature; liquid; molecular weight; nonhuman; pha gene; productivity; repressor gene; volumetry; Wautersia eutropha; whey; Agriculture; Animals; Azotobacter; DNA Primers; Escherichia coli; Hydroxybutyrates; Industrial Waste; Kinetics; Milk; Plasmids; Polyesters; Recombination, Genetic; Zea mays; Azotobacter; Cupriavidus necator; Escherichia coli; Zea mays
Año:2006
Volumen:72
Número:6
Página de inicio:3949
Página de fin:3954
DOI: http://dx.doi.org/10.1128/AEM.00044-06
Título revista:Applied and Environmental Microbiology
Título revista abreviado:Appl. Environ. Microbiol.
ISSN:00992240
CODEN:AEMID
CAS:carbon, 7440-44-0; lactose, 10039-26-6, 16984-38-6, 63-42-3, 64044-51-5; nitrogen, 7727-37-9; poly(3 hydroxybutyric acid), 26063-00-3; DNA Primers; Hydroxybutyrates; Industrial Waste; poly-beta-hydroxybutyrate, 26063-00-3; Polyesters
PDF:https://bibliotecadigital.exactas.uba.ar/download/paper/paper_00992240_v72_n6_p3949_Nikel.pdf
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00992240_v72_n6_p3949_Nikel

Referencias:

  • Ahn, W.S., Park, S.J., Lee, S.Y., Production of poly(3-hydroxybutyrate) by fed-batch culture of recombinant Escherichia coli with a highly concentrated whey solution (2000) Appl. Environ. Microbiol., 66, pp. 3624-3627
  • Aldor, I.S., Keasling, J.D., Process design for microbial plastic factories: Metabolic engineering of polyhydroxyalkanoates (2003) Curr. Opin. Biotechnol., 14, pp. 475-483
  • Bandrup, J., Immergut, E.H., (1989) Polymer Handbook, 3rd Ed., , Interscience Publishers, New York, N.Y
  • Barham, P.G., Keller, A., Otum, E.L., Holmes, P.A., Crystallization and morphology of a bacterial thermoplastic: Poly-β-hydroxybutyrate (1984) J. Mater. Sci., 19, pp. 2781-2794
  • Braunegg, G., Sonnleitner, B., Lafferty, R.M., A rapid gas chromatographic method for the determination of poly-β-hydroxybutyric acid in bacterial biomass (1978) Eur. J. Appl. Microbiol. Biotechnol., 6, pp. 29-37
  • Choi, J.I., Lee, S.Y., Han, K., Cloning of the Alcaligenes latus polyhydroxyalkanoate biosynthesis genes and use of these genes for enhanced production of poly(3-hydroxybutyrate) in Escherichia coli (1998) Appl. Environ. Microbiol., 64, pp. 4897-4903
  • Dawes, E.A., Senior, P.J., The role and regulation of energy reserve polymers in micro-organisms (1973) Adv. Microb. Physiol., 10, pp. 135-266
  • Ditta, G., Schmidhauser, T., Yakobson, E., Lu, P., Liang, X.W., Finlay, D.R., Guiney, D., Helinski, D.R., Plasmids related to the broad host range vector, pRK290, useful for gene cloning and for monitoring gene expression (1985) Plasmid, 13, pp. 149-153
  • Hahn, S.K., Chang, Y.K., Lee, S.Y., Recovery and characterization of poly(3-hydroxybutyric acid) synthesized in Alcaligenes eutrophus and recombinant Escherichia coli (1995) Appl. Environ. Microbiol., 61, pp. 34-39
  • Hanahan, D., Techniques for transformation of Escherichia coli (1985) DNA Cloning: A Practical Approach, 1, pp. 109-135. , D.M. Glover (ed.). IRL Press, Oxford, England
  • Khanna, S., Srivastava, A.K., Recent advances in microbial polyhydroxyalkanoates (2005) Process Biochem., 40, pp. 607-619
  • Kidwell, J., Valentin, H.E., Dennis, D., Regulated expression of the Alcaligenes eutrophus pha biosynthesis genes in Escherichia coli (1995) Appl. Environ. Microbiol., 61, pp. 1391-1398
  • Kim, J.-H., Lee, J., Cloning, nucleotide sequence and expression of gene coding for poly-3-hydroxybutyric acid (PHB) synthase of Rhodobacter sphaeroides 2.4.1 (1997) J. Microbiol. Biotechnol., 7, pp. 229-236
  • Langenbach, S., Rehm, B.H., Steinbüchel, A., Functional expression of the PHA synthase gene phaC1 from Pseudomonas aeruginosa in Escherichia coli results in poly(3-hydroxyalkanoate) synthesis (1997) FEMS Microbiol. Lett., 150, pp. 303-309
  • Lee, S.Y., High cell-density culture of Escherichia coli (1996) Trends Biotechnol., 14, pp. 98-105
  • Lenz, R.W., Marchessault, R.H., Bacterial polyesters: Biosynthesis, biodegradable plastics and biotechnology (2005) Biomacromolecules, 6, pp. 1-8
  • Liu, S.J., Steinbüchel, A., A novel genetically engineered pathway for synthesis of poly(hydroxyalkanoic acids) in Escherichia coli (2000) Appl. Environ. Microbiol., 66, pp. 739-743
  • Madison, L.L., Huisman, G.W., Metabolic engineering of poly(3-hydroxyalkanoates): From DNA to plastic (1999) Microbiol. Mol. Biol. Rev., 63, pp. 21-53
  • Mahishi, L.H., Tripathi, G., Rawal, S.K., Poly(3-hydroxybutyrate) (PHB) synthesis by recombinant Escherichia coli harbouring Streptomyces aureofaciens PHB biosynthesis genes: Effect of various carbon and nitrogen sources (2003) Microbiol. Res., 158, pp. 19-27
  • Nikel, P.I., Pettinari, M.J., Galvagno, M.A., Méndez, B.S., Poly(3-hydroxybutyrate) synthesis by recombinant Escherichia coli arcA mutants in microaerobiosis (2006) Appl. Environ. Microbiol., 72, pp. 2614-2620
  • Nikel, P.I., Pettinari, M.J., Méndez, B.S., Galvagno, M.A., Statistical optimization of a culture medium for biomass and poly(3-hydroxybutyrate) production by a recombinant Escherichia coli strain using agroindustrial by-products (2005) Int. Microbiol., 8, pp. 243-250
  • Ostle, A.G., Holt, J.G., Nile blue A as a fluorescent stain for poly-β-hydroxybutyrate (1982) Appl. Environ. Microbiol., 44, pp. 238-241
  • Pettinari, J.M., Chanetón, L., Vázquez, G., Steinbüchel, A., Méndez, B.S., Insertion sequence-like elements associated with putative polyhydroxybutyrate regulatory genes in Azotobacter sp. FA8 (2003) Plasmid, 50, pp. 36-44
  • Pettinari, M.J., Vázquez, G.J., Silberschmidt, D., Rehm, B., Steinbüchel, A., Méndez, B.S., Poly(3-hydroxybutyrate) synthesis genes in Azotobacter sp. strain FA8 (2001) Appl. Environ. Microbiol., 67, pp. 5331-5334
  • Quagliano, J.C., Alegre, P., Miyazaki, S.S., Isolation and characterization of Azotobacter sp. for the production of poly-β-hydroxyalkanoates (1994) Rev. Argent. Microbiol., 26, pp. 21-27
  • Resch, S., Gruber, K., Wanner, G., Slater, S., Dennis, D., Lubitz, W., Aqueous release and purification of poly(β-hydroxybutyrate) from Escherichia coli (1998) J. Biotechnol., 65, pp. 173-182
  • Sambrook, J., Fritsch, E.F., Maniatis, T., (1988) Molecular Cloning: A Laboratory Manual, 2nd Ed., , Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y
  • Savenkova, L., Gercberga, Z., Nikolaeva, V., Dzene, A., Bibers, I., Kalnin, M., Mechanical properties and biodegradation characteristics of PHB-based films (2000) Process Biochem., 35, pp. 573-579
  • Schlegel, H.G., Lafferty, R., Krauss, I., The isolation of mutants not accumulating poly-β-hydroxybutyric acid (1970) Arch. Microbiol., 71, pp. 283-294
  • Schubert, P., Steinbüchel, A., Schlegel, H.G., Cloning of the Alcaligenes eutrophus genes for synthesis of poly(β-hydroxybutyric acid) (PHB) and synthesis of PHB in Escherichia coli (1988) J. Bacteriol., 170, pp. 5837-5847
  • Shi, H., Kyuwa, K., Takasu, M., Shimizu, K., Temperature-induced expression of phb genes in Escherichia coli and the effect of temperature patterns on the production of poly-3-hydroxybutyrate (2001) J. Biosci. Bioeng., 91, pp. 21-26
  • Shiloach, J., Fass, R., Growing E. coli to high cell density - A historical perspective on method development (2005) Biotechnol. Adv., 23, pp. 345-357
  • Simon, R., Priefer, U., Pühler, A., A broad host range mobilization system for in vivo genetic engineering: Transposon mutagenesis in gram negative bacteria (1983) Bio/Technology, 1, pp. 784-791
  • Steinbüchel, A., PHB and other polyhydroxyalkanoic acids (1995) Biotechnology: Products of Primary Metabolism, 1st Ed., 6, pp. 432-434. , G. R. H. J. Rehm, A. Pühler, and P. Stadler (ed.). Wiley-VCH, Weinheim, Germany
  • Steinbüchel, A., Füchtenbusch, B., Bacterial and other biological systems for polyester production (1998) Trends Biotechnol., 16, pp. 419-427
  • Vandamme, P., Coenye, T., Taxonomy of the genus Cupriavidus: A tale of lost and found (2004) Int. J. Syst. Evol. Microbiol., 54, pp. 2285-2289

Citas:

---------- APA ----------
Nikel, P.I., De Almeida, A., Melillo, E.C., Galvagno, M.A. & Pettinari, M.J. (2006) . New recombinant Escherichia coli strain tailored for the production of poly(3-hydroxybutyrate) from agroindustrial by-products. Applied and Environmental Microbiology, 72(6), 3949-3954.
http://dx.doi.org/10.1128/AEM.00044-06
---------- CHICAGO ----------
Nikel, P.I., De Almeida, A., Melillo, E.C., Galvagno, M.A., Pettinari, M.J. "New recombinant Escherichia coli strain tailored for the production of poly(3-hydroxybutyrate) from agroindustrial by-products" . Applied and Environmental Microbiology 72, no. 6 (2006) : 3949-3954.
http://dx.doi.org/10.1128/AEM.00044-06
---------- MLA ----------
Nikel, P.I., De Almeida, A., Melillo, E.C., Galvagno, M.A., Pettinari, M.J. "New recombinant Escherichia coli strain tailored for the production of poly(3-hydroxybutyrate) from agroindustrial by-products" . Applied and Environmental Microbiology, vol. 72, no. 6, 2006, pp. 3949-3954.
http://dx.doi.org/10.1128/AEM.00044-06
---------- VANCOUVER ----------
Nikel, P.I., De Almeida, A., Melillo, E.C., Galvagno, M.A., Pettinari, M.J. New recombinant Escherichia coli strain tailored for the production of poly(3-hydroxybutyrate) from agroindustrial by-products. Appl. Environ. Microbiol. 2006;72(6):3949-3954.
http://dx.doi.org/10.1128/AEM.00044-06