[논문]Cloning, Characterization of Pichia etchellsii <tex>$\\beta-Glucosidase$</tex> II and Effect of Media Composition and Feeding Strategy on its Production in a Bioreactor

Sethi Benu; Jain Monika; Chowdhary Manish; Soni Yogesh; Bhatia Yukti; Sahai Vikram; Mishra Saroj

doi:10.1007/bf02935879

Cloning, Characterization of Pichia etchellsii $\\beta-Glucosidase$ II and Effect of Media Composition and Feeding Strategy on its Production in a Bioreactor 원문보기

Biotechnology and bioprocess engineering : Bbe, v.7 no.1, 2002년, pp.43 - 51

Sethi Benu (Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology) , Jain Monika (Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology) , Chowdhary Manish (Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology) , Soni Yogesh (Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology) , Bhatia Yukti (Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology) , Sahai Vikram (Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology) , Mishra Saroj (Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology)

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The cloning and expression of $\beta-glucosidase$ II, encoded by the gene ${\beta}glu2$, from thermotolerant yeast Pichia etchellsii into Escherichia coli is described. Cloning of the 7.3 kb BamHI/SalI yeast insert containing ${\beta}glu2$ in pUC18, which allowed for reverse orientation of the insert, resulted in better enzyme expression. Transformation of this plasmid into E. coli JM109 resulted in accumulation of the enzyme in periplasmic space. At $50^{\circ}C$, the highest hydrolytic activity of 1686 IU/g protein was obtained on sophorose. Batch and fed-batch techniques were employed for enzyme production in a 14 L bioreactor. Exponential feeding rates were determined from mass balance equations and these were employed to control specific growth rate and in turn maximize cell growth and enzyme production. Media optimization coupled with this strategy resulted in increased enzyme units of 1.2 kU/L at a stabilized growth rate of $0.14\;h^{-l}$. Increased enzyme production in bioreactor was accompanied by formation of inclusion bodies.

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1 U/g) where no IPTG was added. In view of the multi copy nature of pUC₁₈ in which all lac regulatory sites may not be titrated out by the level of repressor in the uninduced culture (and thereby mask the difference between the two experimental conditions, uninduced and induced), an attempt was made to titrate out the repressor by co-transformation with plasmid pAH₃ (for description, see Materials and Methods) and selecting the transformants on LB + amp and kanamycin. However; stable transformants were not obtained.

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coli DH₅이 13] and JM109 [14]. Plasmids pUC₁₉ and pUC₁₈ were obtained from Bangalore Geneij Bangalore. Plasmid pAH₃ [15] was a kind gift from Dr.
The yeast Pichia etchellsii (Deutsche Sammlung Von Mikrorganismen (DSM) Germany) was used as the donor of the P-glucosidase gene. The bacterial strains used in this study were E. coli DH₅이 13] and JM109 [14]. Plasmids pUC₁₉ and pUC₁₈ were obtained from Bangalore Geneij Bangalore.
Southern hybridisation to determine the origin of /3glu2 from genomic DNA of P etchellsii was performed. The data showed hybridisation signals with a single 8.5 kb fragment of chromosomal DNA. Dot-blot hybridisation experiments were also performed to examine whether DNA sequences from 2.

참고문헌 (36)

CRC Crit. Rev. Biotechnol. V. S. Bisaria 9 61 1989 10.3109/07388558909040616 Bisaria, V. S. and S. Mishra (1989) Regulatory aspects of cellulase biosynthesis and secretion.CRC Crit. Rev. Biotechnol. 9: 61-103.

상세보기
Enzyme Microb. Technol. C. P. Kubicek 15 90 1993 10.1016/0141-0229(93)90030-6 Kubicek, C. P., R. Messner, F. Gruber, R. L. Mach, and E. M. Kubicek-Pranz (1993) TheTrichoderma cellulase regulatory puzzle: From the interior life of a secretory fungus.Enzyme Microb. Technol. 15: 90-99.

상세보기
Biotechnol. Bioprocess Eng. B. Sethi 4 189 1999 10.1007/BF02931927 Sethi, B., S. Mishra, and V. S. Bisaria (1999) Sorbose mediated enhancement of cellulase biosynthesis inTrichoderma ressei.Biotechnol. Bioprocess Eng. 4: 189-194.

상세보기
A. Esen 1993 10.1021/bk-1993-0533 ACS symposium series β-Glucosidases, Biochemistry and Molecular biolog Esen, A. (1993) β-Glucosidases, Biochemistry and Molecular biology, ACS symposium series, ACS Washington, DC, USA.
D. S. Fredrickson 730 1972 The metabolic basis of inherited disease 3rd ed. Fredrickson, D. S., and M. R. Sloan (1972) The pathogenicity of gaucher's disease. pp. 730-745. In: J. B. Stanbury, J. B. Wyngaarden, and D. S. Fredrickson (eds.), The metabolic basis of inherited disease. 3rd ed. McGraw-Hill, NY, USA.
Biotechnol. Lett. K. Ajisaka 9 243 1987 10.1007/BF01027157 Ajisaka, K., H. Nishida, and H. Fujimoto (1987) The synthesis of oligosaccharides by the reverse hydrolysis reaction of β-glucosidase at high substrate concentration and at high temperature.Biotechnol. Lett. 9: 243-248.

상세보기
Tetrahedron Lett. G. Vic 33 4567 1992 10.1016/S0040-4039(00)61314-X Vic, G. and D. Thomas (1992) Enzyme catalysed synthesis of alkyl-D-glucosides in organic media.Tetrahedron Lett. 33: 4567-4577.

상세보기
Biotechnol. Lett. L. Fischer 17 1169 1995 10.1007/BF00128380 Fischer, L., R. Bromann, and F. Wagner (1995) Enantioselective synthesis of several 1-O-D-glycoconjugates using almond β-glucosidase (EC 3.2.1.21).Biotechnol. Lett. 17: 1169-1174.

상세보기
J. Ferment. Bioeng. M. Pandey 80 446 1995 10.1016/0922-338X(96)80918-4 Pandey, M. and S. Mishra (1995) Cloning and expression of β-glucosidase gene from the yeastPichia etchellsii.J. Ferment. Bioeng. 80: 446-453.

상세보기
Gene M. Pandey 190 45 1997 10.1016/S0378-1119(96)00712-3 Pandey, M. and S. Mishra (1997) Expression and characterization ofPichia etchellsii β-glucosidase inEscherichia coli.Gene 190: 45-51.

상세보기
Methods: A Companion to Methods in Enzymology N. S. Lin 4 159 1992 10.1016/1046-2023(92)90048-D Lin, N. S. and J. R. Swartz (1992) Production of heterologous proteins from recombinant DNAEscherichia coli in bench fermentors.Methods: A Companion to Methods in Enzymology 4: 159-170.

상세보기
Bio/Technology L. Yee 10 1550 1993 Yee, L. and H. W. Blanch (1993) Recombinant protein expression in high cell density fed batch cultures ofEschrrichia coli.Bio/Technology 10: 1550-1556.
E. A. Raleigh 1989 Current Protocols in Molecular Biology Raleigh, E. A., K. Lech, and R. Brent (1989) Selected topics from classical bacterial genetics. Unit 1.4. In F. M. Ausubel, R. Brent, R. E. Kingston, D. D. Moore J. G. Seidman, J. A. Smith, and K. Struhl (eds.).Current Protocols in Molecular Biology. Publishing associates and Wiley Interscience, NY, USA.
Gene C. Yanisch-Perron 33 103 1985 10.1016/0378-1119(85)90120-9 Yanisch-Perron, C., J. Vieira, and J. Messing (1985) Improvement in M13 phage cloning vectors and host strains: nucleotide sequences of the M13m18 and pUC19 vectors.Gene 33: 103-119.

상세보기
Nucleic Acids Res. A. Higashitani 20 2685 1992 10.1093/nar/20.11.2685 Higashitani, A., D. Greenstein, and K. Horiuchi (1992) A single amino acid substitution reduces the super helicity requirement of a replication initiator protein.Nucleic Acids Res. 20: 2685-2691.

상세보기
Mol. Cell Biol. J. M. Cregg 5 3376 1985 10.1128/MCB.5.12.3376 Cregg, J. M., K. J. Barringer, A. Y Hessler, and K. R. Madden (1985)Pichia pastoris as a host system for transformations.Mol. Cell Biol. 5: 3376-3385.

상세보기
J. Sambrook 1.21 1989 Molecular Cloning: A Laboratory Manual 2nd ed. Sambrook, J., E. F. Fritsch, and T. Maniatis (1989)Molecular Cloning: A Laboratory Manual, 2nd ed., pp. 1.21-1.85. Cold Spring Harbor Laboratory Press, NY, USA.
J. Biol. Chem. N. G. Nossal 241 3055 1966 10.1016/S0021-9258(18)96497-5 Nossal, N. G. and L. A. Heppel (1966) The release of enzymes by osmotic shock fromEscherichia coli in exponential phase.J. Biol. Chem. 241: 3055-3062.

상세보기
S. Gallagher 1993 Current Protocols in Molecular Biology Gallagher, S., S. E. Winston, S. A. Fuller, and J. G. R. Huriell (1993) Analysis of Proteins. Unit 10.8 In: F. M. Ausubel, R. Brent, R. E. Kingston, D. D. Moore, J. G. Seidman, J. A. Smith, and K. Struhl (eds.).Current Protocols in Molecular Biology. Publishing Associates and Wiley Interscience, NY, USA.
S. J. Pirt 42 1975 Principles of Microbe and Cell Cultivation Pirt, S. J. (1975)Principles of Microbe and Cell Cultivation. pp. 42-44. Blackwell Scientific, Oxford, UK.
J. Forment. Bioeng. S. Shima 68 75 1989 10.1016/0922-338X(89)90050-0 Shima, S., J. Kato, Y. igarashi, and T. Kodama (1989) Cloning and expression of aClostridium cellobioparum cellulase gene and its expression inEscherichia coli JM109.J. Forment. Bioeng. 68: 75-78.

상세보기
J. Bacteriol. E. Painbeni 174 3087 1992 10.1128/jb.174.9.3087-3091.1992 Painbeni, E., S. Valles, J. Polaina, and A. Flors (1992) Purification and characterisation of aBacillus polymxa Bgl gene expressed inEscherichia coli.J. Bacteriol. 174: 3087-3091.

상세보기
J. Bacteriol. W. G. B. Voorhorst 177 7105 1995 10.1128/jb.177.24.7105-7111.1995 Voorhorst, W. G. B., R. I. L. Eggen, E. J. Leusink, and de W. M. Vos (1995) Characterization ofcelB gene encoding for β-glucosidase from the hypothermophilic archeonPyrococcus furiosus and its expression and site-directed mutation inEscherichia coli.J. Bacteriol. 177: 7105-7111.

상세보기
Process Biochem. D.-H. Kweon 36 537 2001 10.1016/S0032-9592(00)00237-5 Kweon, D.-H., N. S. Han, K.-M. Park, and J.-H. Seo (2001) Overproduction ofPhytolacca insularis protein in batch and fed-batch culture of recombinantEscherichia coli.Process Biochem. 36: 537-542.

상세보기
FEMS Microbiol. Lett. L. Marri 128 135 1995 Marri, L., S. Valentini, and D. Venditti (1995) Cloning and nucleotide sequence ofbglA fromErwinia herbicola and expression of bgl activity inEscherichia coli.FEMS Microbiol. Lett. 128: 135-138.

상세보기
Mol. Gen. Genet. S. Vroeman 246 465 1995 10.1007/BF00290450 Vroeman, S., J. Heldens, C. Boyd, B. Henrisaat, and N. T. Keen (1995) Cloning and characterization ofbgxa gene fromErwinia chrysanthemi D1 which encodes a β-glucosidase/ xylosidase enzyme.Mol. Gen. Genet. 246: 465-477.

상세보기
Appl. Environ. Microbiol. R. Breves 63 3902 1997 10.1128/AEM.63.10.3902-3910.1997 Breves, R., K. Bronnenmeier, N. Wild, F. Lottspeich, W. L. Staudenbauer, and J. Hofemeister (1997) Genes encoding two different β-glucosidases ofThermoanaerobacter brokii are clustered in a common operon.Appl. Environ. Microbiol. 63: 3902-3910.

상세보기
Appl. Microbiol. Biotechnol. D. Reisenberg 51 422 1999 10.1007/s002530051412 Reisenberg, D. and R. Guthke (1999) High cell density cultivation of microorganisms.Appl. Microbiol. Biotechnol. 51: 422-430.

상세보기
Biotechnol. Bioeng. B. Jenson 36 1 1990 10.1002/bit.260360102 Jenson, B. and S. Carlsen (1990) Production of recombinant human growth hormone inEscherichia coli: Expression of different precursors and physiological effects of glucose, acetate and salts.Biotechnol. Bioeng. 36: 1-11.

상세보기
Trends Biotechnol. S. Y. Lee 14 98 1996 10.1016/0167-7799(96)80930-9 Lee, S. Y. (1996) High cell-density culture ofEscherichia coli.Trends Biotechnol. 14: 98-105.

상세보기
Biotechnol. Prog. W. Srittmatter 13 247 1997 Srittmatter, W. and S. Matzku (1997) Production of heterologous protein inEscherichia coli under high cell density conditions.Biotechnol. Prog. 13: 247-257.
Biotechnol. Bioeng. S. K. Yoon 43 995 1994 10.1002/bit.260431013 Yoon, S. K., W. K. Kang, and T. H. Park (1994) Fed-batch operation of recombinantEscherichia coli containingtrp promoter with controlled specific growth rate.Biotechnol. Bioeng. 43: 995-999.

상세보기
J. Biotechnol. D. J. Korz 39 59 1995 10.1016/0168-1656(94)00143-Z Korz, D. J., V. Rinas, K. Hellmuth, E. A. Sanders, and W. D. Deckwer (1995) Simple fed-batch technique for high cell density cultivation ofEscherichia coli.J. Biotechnol. 39: 59-65 (1995).

상세보기
Biotechnol. Prog. C. H. Chou 10 644 1994 10.1021/bp00030a009 Chou, C. H., G. N. Bennet, and K. Y. San (1994) Effect of modulated glucose uptake on high level recombinant protein production in a denseEscherichia coli culture.Biotechnol. Prog. 10: 644-647.

상세보기
J. Biotechnol. M. Schmidt 68 71 1999 10.1016/S0168-1656(98)00189-8 Schmidt, M., K. R. Babu, N. Khanna, S. Marten, and U. Rinas (1999) Temperature induced production of recombinant human insulin in high-cell density cultures of recombinantEscherichia coli.J. Biotechnol. 68: 71-83.

상세보기
Biotechnol. Bioeng. C. Schiraldi 70 670 2000 10.1002/1097-0290(20001220)70:6<670::AID-BIT9>3.0.CO;2-7 Schiraldi, C., A. Martino, M. Acone, I. Di Lernia, A. Di Lazzaro, F. Marulli, M. Generoso, M. Carteni, and M. De Rosa (2000) Effective production of a thermostable α-glucosidase fromSulfolobus solfataricus inEscherichia coli exploiting a microfiltration bireactor.Biotechnol. Bioeng. 70: 670-676.

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