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Abstract:

Several biotechnological processes rely on the utilization of high-copy-number plasmids for heterologous gene expression, and understanding the interactions between plasmid DNA and bacterial hosts is highly relevant for bioprocess optimization. We assessed metabolic modifications and physiological changes exerted by expression of a plasmid-encoded alcohol-acetaldehyde dehydrogenase from Leuconostoc mesenteroides (adhE Lm ) in Escherichia coli redox mutants. Plasmid pET Lm, a pBluescript II KS(-)-derivative carrying adhE Lm, was introduced in E. coli CT1061 [arcA creC(Con)]. This recombinant was able to attain a higher ethanol concentration in glycerol cultures compared to the parental strain. pBluescript II KS(-) was rapidly lost in 72-h bioreactor cultures (7.8±1.2% of plasmid-bearing cells), while pET Lm was present in 92.4±7.2% of the cells. In E. coli CT1061 carrying pBluescript II KS(-) the plasmid copy number steadily diminished in bioreactor cultures to reach 334±45 copies per chromosome at 72 h, while pET Lm was stably maintained, reaching 498±18 copies per chromosome at the end of the cultivation. Plasmid pETΩ Lm, bearing a defective copy of adhE Lm interrupted by cat, reached 293±62 copies per chromosome, implying a functional role of adhE Lm on plasmid maintenance. The intracellular NADH/NAD+ content suggest that regeneration of oxidized co-factors by the heterologous bioreaction might play a relevant role in plasmid maintenance. © 2010 Springer-Verlag.

Registro:

Documento: Artículo
Título:Metabolic selective pressure stabilizes plasmids carrying biosynthetic genes for reduced biochemicals in Escherichia coli redox mutants
Autor:Nikel, P.I.; Pettinari, M.J.; Galvagno, M.A.; Méndez, B.S.
Filiación:Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Av. Intendente Güiraldes 2160, C1428EGA Ciudad Autónoma de Buenos Aires, Argentina
Instituto de Investigaciones Biotecnológicas, Universidad Nacional de San Martín, Av. General Paz 5445, San Martín, Buenos Aires B1650KNA, Argentina
Departamento de Ingeniería Química, Facultad de Ingeniería, Universidad de Buenos Aires, Av. Intendente Güiraldes 2160, C1428EGA Ciudad Autónoma de Buenos Aires, Argentina
Palabras clave:ArcAB; CreBC; Escherichia coli; Microaerobiosis; Plasmid copy number; Redox mutants; ArcAB; Copy number; CreBC; Microaerobiosis; Redox mutants; Aldehydes; Bearings (structural); Bioconversion; Bioreactors; Cell culture; Chromosomes; Escherichia coli; Ethanol; Gene expression; Glycerol; Maintenance; Metabolism; Organic polymers; Microbiology; alcohol; alcohol acetaldehyde dehydrogenase; aldehyde dehydrogenase; glycerol; nicotinamide adenine dinucleotide; reduced nicotinamide adenine dinucleotide; unclassified drug; biochemistry; bioreactor; chromosome; coliform bacterium; DNA; enzyme; ethanol; gene expression; metabolism; mutagenicity; plasmid; article; bacterial chromosome; bacterial gene; bacterium culture; bioreactor; biosynthesis; controlled study; Escherichia coli; Leuconostoc mesenteroides; mutant; nonhuman; oxidation; oxidation reduction reaction; physiology; plasmid; Alcohol Dehydrogenase; Aldehyde Oxidoreductases; Escherichia coli; Leuconostoc; Mutation; Oxidation-Reduction; Plasmids; Selection, Genetic; Arca; Bacteria (microorganisms); Escherichia coli; Leuconostoc mesenteroides
Año:2010
Volumen:88
Número:2
Página de inicio:563
Página de fin:573
DOI: http://dx.doi.org/10.1007/s00253-010-2774-1
Título revista:Applied Microbiology and Biotechnology
Título revista abreviado:Appl. Microbiol. Biotechnol.
ISSN:01757598
CODEN:AMBID
CAS:alcohol, 64-17-5; aldehyde dehydrogenase, 37353-37-0, 9028-86-8; glycerol, 56-81-5; nicotinamide adenine dinucleotide, 53-84-9; reduced nicotinamide adenine dinucleotide, 58-68-4; Alcohol Dehydrogenase, 1.1.1.1; Aldehyde Oxidoreductases, 1.2.-; acetaldehyde dehydrogenase, 1.2.1.5
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01757598_v88_n2_p563_Nikel

Referencias:

  • Avison, M.B., Horton, R.E., Walsh, T.R., Bennett, P.M., Escherichia coli CreBC is a global regulator of gene expression that responds to growth in minimal media (2001) J Biol Chem, 276, pp. 26955-26961. , 10.1074/jbc.M011186200
  • Bai, F.W., Anderson, W.A., Moo-Young, M., Ethanol fermentation technologies from sugar and starch feedstocks (2008) Biotechnol Adv, 26, pp. 89-105. , 10.1016/j.biotechadv.2007.09.002
  • Bentley, W.E., Mirjalili, N., Andersen, D.C., Davis, R.H., Kompala, D.S., Plasmid-encoded protein: The principal factor in the "metabolic burden" associated with recombinant bacteria (1990) Biotechnol Bioeng, 35, pp. 668-681. , 10.1002/bit.260350704
  • Bernofsky, C., Swan, M., An improved cycling assay for nicotinamide adenine dinucleotide (1973) Anal Biochem, 53, pp. 452-458. , 10.1016/0003-2697(73)90094-8
  • Bower, D.M., Prather, K.L.J., Engineering of bacterial strains and vectors for the production of plasmid DNA (2009) Appl Microbiol Biotechnol, 82, pp. 805-813. , 10.1007/s00253-009-1889-8
  • Bradford, M.M., A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding (1976) Anal Biochem, 72, pp. 248-254. , 10.1016/0003-2697(76)90527-3
  • Clomburg, J.M., Gonzalez, R., Biofuel production in Escherichia coli: The role of metabolic engineering and synthetic biology (2010) Appl Microbiol Biotechnol, 86, pp. 419-434. , 10.1007/s00253-010-2446-1
  • Cranenburgh, R.M., Hanak, J.A.J., Williams, S.G., Sherratt, D.J., Escherichia coli strains that allow antibiotic-free plasmid selection and maintenance by repressor titration (2001) Nucleic Acids Res, 29, p. 26. , 10.1093/nar/29.5.e26
  • Da Silva, G.P., MacK, M., Contiero, J., Glycerol: A promising and abundant carbon source for industrial microbiology (2009) Biotechnol Adv, 27, pp. 30-39. , 10.1016/j.biotechadv.2008.07.006
  • Datsenko, K., Wanner, B.L., One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products (2000) Proc Natl Acad Sci USA, 97, pp. 6640-6645. , 10.1073/pnas.120163297
  • Demain, A.L., Biosolutions to the energy problem (2009) J Ind Microbiol Biotechnol, 36, pp. 319-332. , 10.1007/s10295-008-0521-8
  • Díaz-Ricci, J.C., Hernández, M.E., Plasmid effects on Escherichia coli metabolism (2000) Crit Rev Biotechnol, 20, pp. 79-108. , 10.1080/07388550008984167
  • Díaz-Ricci, J.C., Bode, J., Rhee, J.I., Schügerl, K., Gene expression enhancement due to plasmid maintenance (1995) J Bacteriol, 177, pp. 6684-6687
  • Dumsday, G.J., Zhou, B., Yaqin, W., Stanley, G.A., Pamment, N.B., Comparative stability of ethanol production by Escherichia coli KO11 in batch and chemostat cultures (1999) J Ind Microbiol Biotechnol, 23, pp. 701-708. , 10.1038/sj.jim.2900690
  • Durany, O., Bassett, P., Weiss, A.M.E., Cranenburgh, R.M., Ferrer, P., López-Santín, J., De Mas, C., Hanak, J.A.J., Production of fuculose-1-phosphate aldolase using operator-repressor titration for plasmid maintenance in high cell density Escherichia coli fermentations (2005) Biotechnol Bioeng, 91, pp. 460-467. , 10.1002/bit.20527
  • Friehs, K., Plasmid copy number and plasmid stability (2004) Adv Biochem Eng Biotechnol, 86, pp. 47-82
  • Gibon, Y., Larher, F., Cycling assay for nicotinamide adenine dinucleotides: NaCl precipitation and ethanol solubilization of the reduced tetrazolium (1997) Anal Biochem, 251, pp. 153-157. , 10.1006/abio.1997.2283
  • Gokarn, R.R., Evans, J.D., Walker, J.R., Martin, S.A., Eiteman, M.A., Altman, E., The physiological effects and metabolic alterations caused by the expression of Rhizobium etli pyruvate carboxylase in Escherichia coli (2001) Appl Microbiol Biotechnol, 56, pp. 188-195. , 10.1007/s002530100661
  • Grabherr, R., Bayer, K., Impact of targeted vector design on ColE1 plasmid replication (2002) Trends Biotechnol, 20, pp. 257-260. , 10.1016/S0167-7799(02)01950-9
  • Hespell, R.B., Wyckoff, H., Dien, B.S., Bothast, R.J., Stabilization of pet operon plasmids and ethanol production in Escherichia coli strains lacking lactate dehydrogenase and pyruvate formate-lyase activities (1996) Appl Environ Microbiol, 62, pp. 4594-4597
  • Ingram, L.O., Conway, T., Clark, D.P., Sewell, G.W., Preston, J.F., Genetic engineering of ethanol production in Escherichia coli (1987) Appl Environ Microbiol, 53, pp. 2420-2425
  • Jarboe, L.R., Grabar, T.B., Yomano, L.P., Shanmugam, K.T., Ingram, L.O., Development of ethanologenic bacteria (2007) Adv Biochem Eng Biotechnol, 108, pp. 237-261
  • Koo, O.K., Jeong, D.W., Lee, J.M., Kim, M.J., Lee, J.H., Chang, H.C., Kim, J.H., Lee, H.J., Cloning and characterization of the bifunctional alcohol/acetaldehyde dehydrogenase gene (adhE) in Leuconostoc mesenteroides isolated from kimchi (2005) Biotechnol Lett, 27, pp. 505-510. , 10.1007/s10529-005-2541-z
  • Kroll, J., Steinle, A., Reichelt, R., Ewering, C., Steinbüchel, A., Establishment of a novel anabolism-based addiction system with an artificially introduced mevalonate pathway: Complete stabilization of plasmids as universal application in white biotechnology (2009) Metab Eng, 11, pp. 168-177. , 10.1016/j.ymben.2009.01.007
  • Lawford, H.G., Rousseau, J.D., Loss of ethanologenicity in Escherichia coli B recombinants pLOI297 and KO11 during growth in the absence of antibiotics (1995) Biotechnol Lett, 17, pp. 751-756. , 10.1007/BF00130363
  • Lawford, H.G., Rousseau, J.D., Factors contributing to the loss of ethanologenicity of Escherichia coli B recombinants pLOI297 and KO11 (1996) Appl Biochem Biotechnol, 5758, pp. 293-305. , 10.1007/BF02941709
  • MacKenzie, K.F., Eddy, C.K., Ingram, L.O., Modulation of alcohol dehydrogenase isoenzyme levels in Zymomonas mobilis by iron and zinc (1989) J Bacteriol, 171, pp. 1063-1067
  • Neidhardt, F.C., Ingraham, J.L., Schaechter, M., (1990) Physiology of the Bacterial Cell: A Molecular Approach, , Sinauer Sunderland
  • Nielsen, J., Villadsen, J., Lidén, G., (2003) Bioreaction Engineering Principles, , 2 Kluwer/Plenum New York
  • 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. , 10.1128/AEM.72.4.2614-2620.2006
  • Nikel, P.I., De Almeida, A., Pettinari, M.J., Méndez, B.S., The legacy of HfrH: Mutations in the two-component system CreBC are responsible for the unusual phenotype of an Escherichia coli arcA mutant (2008) J Bacteriol, 190, pp. 3404-3407. , 10.1128/JB.00040-08
  • Nikel, P.I., Pettinari, M.J., Galvagno, M.A., Méndez, B.S., Poly(3-hydroxybutyrate) synthesis from glycerol by a recombinant Escherichia coli arcA mutant in fed-batch microaerobic cultures (2008) Appl Microbiol Biotechnol, 77, pp. 1337-1343. , 10.1007/s00253-007-1255-7
  • Nikel, P.I., Pettinari, M.J., Ramirez, M.C., Galvagno, M.A., Méndez, B.S., Escherichia coli arcA mutants: Metabolic profile characterization of microaerobic cultures using glycerol as a carbon source (2008) J Mol Microbiol Biotechnol, 15, pp. 48-54. , 10.1159/000111992
  • Nikel, P.I., Zhu, J., San, K.Y., Méndez, B.S., Bennett, G.N., Metabolic flux analysis of Escherichia coli creB and arcA mutants reveals shared control of carbon catabolism under microaerobic growth conditions (2009) J Bacteriol, 191, pp. 5538-5548. , 10.1128/JB.00174-09
  • Nikel, P.I., Ramirez, M.C., Pettinari, M.J., Méndez, B.S., Galvagno, M.A., Ethanol synthesis from glycerol by Escherichia coli redox mutants expressing adhE from Leuconostoc mesenteroides (2010) J Appl Microbiol, 109, pp. 492-504
  • Overath, P., Schairer, H.U., Stoffel, W., Correlation of in vivo and in vitro phase transitions of membrane lipids in Escherichia coli (1970) Proc Natl Acad Sci USA, 67, pp. 606-612. , 10.1073/pnas.67.2.606
  • Ow, D.S.W., Nissom, P.M., Philp, R., Oh, S.K.W., Yap, M.G.S., Global transcriptional analysis of metabolic burden due to plasmid maintenance in Escherichia coli DH5α during batch fermentation (2006) Enzyme Microb Technol, 39, pp. 391-398. , 10.1016/j.enzmictec.2005.11.048
  • Peralta-Yahya, P.P., Keasling, J.D., Advanced biofuel production in microbes (2010) Biotechnol J, 5, pp. 147-162. , 10.1002/biot.200900220
  • Pettinari, M.J., Nikel, P.I., Ruiz, J.A., Méndez, B.S., ArcA redox mutants as a source of reduced bioproducts (2008) J Mol Microbiol Biotechnol, 15, pp. 41-47. , 10.1159/000111991
  • Rozkov, A., Avignone-Rossa, C.A., Ertl, P.F., Jones, P., O'Kennedy, R.D., Smith, J.J., Dale, J.W., Bushell, M.E., Characterization of the metabolic burden on Escherichia coli DH1 cells imposed by the presence of a plasmid containing a gene therapy sequence (2004) Biotechnol Bioeng, 88, pp. 909-915. , 10.1002/bit.20327
  • Sambrook, J., Russell, D.W., (2001) Molecular Cloning: A Laboratory Manual, , 3 Cold Spring Harbor Laboratory Cold Spring Harbor
  • Shiloach, J., Fass, R., Growing E. coli to high cell density-A historical perspective on method development (2005) Biotechnol Adv, 23, pp. 345-357. , 10.1016/j.biotechadv.2005.04.004
  • Singh, A., Lynch, M.D., Gill, R.T., Genes restoring redox balance in fermentation-deficient E. coli NZN111 (2009) Metab Eng, 11, pp. 347-354. , 10.1016/j.ymben.2009.07.002
  • Škulj, M., Okršlar, V., Jalen, Š., Jevševar, S., Slanc, P., Štrukelj, B., Menart, V., Improved determination of plasmid copy number using quantitative real-time PCR for monitoring fermentation processes (2008) Microb Cell Fact, 7, p. 6. , 10.1186/1475-2859-7-6
  • Solomon, B.D., Biofuels and sustainability (2010) Ann NY Acad Sci, 1185, pp. 119-134. , 10.1111/j.1749-6632.2009.05279.x
  • Summers, D., Timing, self-control and a sense of direction are the secrets of multicopy plasmid stability (1998) Mol Microbiol, 29, pp. 1137-1145. , 10.1046/j.1365-2958.1998.01012.x
  • Teich, A., Lin, H.Y., Andersson, L., Meyer, S., Neubauer, P., Amplification of ColE1 related plasmids in recombinant cultures of Escherichia coli after IPTG induction (1998) J Biotechnol, 64, pp. 197-210. , 10.1016/S0168-1656(98)00108-4
  • Vertès, A.A., Inui, M., Yukawa, H., Technological options for biological fuel ethanol (2008) J Mol Microbiol Biotechnol, 15, pp. 16-30. , 10.1159/000111989
  • Wang, Z., Xiang, L., Shao, J., Wȩgrzyn, A., Wȩgrzyn, G., Effects of the presence of ColE1 plasmid DNA in Escherichia coli on the host cell metabolism (2006) Microb Cell Fact, 5, p. 34. , 10.1186/1475-2859-5-34
  • Weber, A.E., San, K.Y., A comparison of two plating techniques to estimate plasmid stability of a prolonged chemostat culture (1989) Biotechniques, 3, pp. 397-400
  • Wȩgrzyn, G., Wȩgrzyn, A., Stress responses and replication of plasmids in bacterial cells (2002) Microb Cell Fact, 1, p. 2. , 10.1186/1475-2859-1-2
  • Yau, S.Y., Keshavarz-Moore, E., Ward, J., Host strain influences on supercoiled plasmid DNA production in Escherichia coli: Implications for efficient design of large-scale processes (2008) Biotechnol Bioeng, 101, pp. 529-544. , 10.1002/bit.21915
  • Zielenkiewicz, U., Cegłowski, P., Mechanisms of plasmid stable maintenance with special focus on plasmid addiction systems (2001) Acta Biochim Pol, 48, pp. 1003-1023

Citas:

---------- APA ----------
Nikel, P.I., Pettinari, M.J., Galvagno, M.A. & Méndez, B.S. (2010) . Metabolic selective pressure stabilizes plasmids carrying biosynthetic genes for reduced biochemicals in Escherichia coli redox mutants. Applied Microbiology and Biotechnology, 88(2), 563-573.
http://dx.doi.org/10.1007/s00253-010-2774-1
---------- CHICAGO ----------
Nikel, P.I., Pettinari, M.J., Galvagno, M.A., Méndez, B.S. "Metabolic selective pressure stabilizes plasmids carrying biosynthetic genes for reduced biochemicals in Escherichia coli redox mutants" . Applied Microbiology and Biotechnology 88, no. 2 (2010) : 563-573.
http://dx.doi.org/10.1007/s00253-010-2774-1
---------- MLA ----------
Nikel, P.I., Pettinari, M.J., Galvagno, M.A., Méndez, B.S. "Metabolic selective pressure stabilizes plasmids carrying biosynthetic genes for reduced biochemicals in Escherichia coli redox mutants" . Applied Microbiology and Biotechnology, vol. 88, no. 2, 2010, pp. 563-573.
http://dx.doi.org/10.1007/s00253-010-2774-1
---------- VANCOUVER ----------
Nikel, P.I., Pettinari, M.J., Galvagno, M.A., Méndez, B.S. Metabolic selective pressure stabilizes plasmids carrying biosynthetic genes for reduced biochemicals in Escherichia coli redox mutants. Appl. Microbiol. Biotechnol. 2010;88(2):563-573.
http://dx.doi.org/10.1007/s00253-010-2774-1