Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Production of Intracellular Invertase from Alkalophilic and Thermophilic Bacillus sp. TA-11 in the Recombinant E. coli

재조합 대장균에서 호알칼리성,고온성 Bacillus sp. TA-11의 세포내 Invertase의 생산

  • Yi, Sung-Hun (Department of Food Science and Agricultural Chemistry, Macdonald College of McGilll University) ;
  • Lee, Dae-Hyung (Department of Life Science and Genetic Engineering, Paichai University) ;
  • No, Jae-Duck (Department of Life Science and Genetic Engineering, Paichai University) ;
  • Lee, Jae-Won (Department of Life Science and Genetic Engineering, Paichai University) ;
  • Lee, Jong-Soo (Department of Life Science and Genetic Engineering, Paichai University)
  • 이성훈 (맥길대학교 식품.농화학과) ;
  • 이대형 (배재대학교 생명유전공학과) ;
  • 노재덕 (배재대학교 생명유전공학과) ;
  • 이재원 (배재대학교 생명유전공학과) ;
  • 이종수 (배재대학교 생명유전공학과)
  • Published : 2006.12.28
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Abstract

The intracellular invertase gene of alkalophilic and thermophilic Bacillus sp. TA-11 which was isolated from compost was cloned and expressed in E. coil HB101 using pUC19 as a vector. The invertase of the recombinant E. coli (pYC 17) was maximally produced when it was incubated at 37$^{\circ}C$ for 9 h in a SY medium containing 0.25% sucrose, 0.5% yeast extract, 0.1% each of $K_2HPO_4$ and $KH_2PO_4$, with an initial pH of 8.0. The final enzyme activity under the above condition was 47.7 U per ml of cell-free extract.

알칼리와 고온에 대한 내성을 가진 invertase를 대량생산하기 위하여 호알칼리성이며 고온성 세균인 Bacillus sp. TA-11의 세포내 invertase유전자를 pUC 19벡터를 이용하여 E. coli HB101에 클로닝 시키고 발현시켜 invertase를 강력하게 생산하는 재조합 E. coli (pYC17)를 얻었다. 재조합 E. coli(pYC17)를 이용한 invertase 생산 최적조건을 검토한 결과 E. coil(pYC17)를 0.25% sucrose, 0.5% yeast extract, 0.1% $K_2HPO_4$와 0.1% $KH_2PO_4$을 함유한 SY배지(초기 pH 9.0)에 접종하여 37$^{\circ}C$에서 9시간 배양하였을 때 친주보다도 많은 47.7 U/ml-cell free extract의 invertase가 생산되었다.

Keywords

References

  1. Abrams, B. B., R. Hckel, T. Mizunaga, and J. O. Lampen, 1978. Relationship of large and small invertase in Saccharomyese; Mutant selectively deficient in small invertase. J. Bacteriol. 135: 809-817
  2. Arnold, W. N. 1972. Location of acid phosphatase and ${\beta}$-fructofuranosidase within yeast cell envelopes. J. Bacteriol. 112: 1346-1352
  3. Babczinski, P. 1980. Partial purification, characterization and localization of the membrane-associated invertase of yeast. Biochim. Biophys. Acta. 614: 121-133
  4. Choi, Y. J. and J. S. Lee. 1995. Production of ${\beta}$-fructofuransosidase from alkalophilic, thermophilic Bacillus sp. TA-11. Korean J. Appl. Microbiol. Biotechnol. 23: 197-202
  5. Fitzgerald, R. J., D. M. Spinell, and T. H. Stoudt. 1968. Enzymatic removal of artificial plague. Arch. Oral. Biol. 13: 125-128 https://doi.org/10.1016/0003-9969(68)90042-3
  6. Dodyk, F. and A. Rothstein. 1964. Factors influencing the apperarance of invertase in Saccharomyces cerevisiae. Arch. Biochem. Biophys. 104: 478-486 https://doi.org/10.1016/0003-9861(64)90492-8
  7. Fouet, A., A. Klier, and G. Rapoport. 1986. Nucleotide sequence of the sucrose gene of Bacillus subtilis. Gene. 45: 221-225 https://doi.org/10.1016/0378-1119(86)90258-1
  8. Fujita, K., K. Hara, H. Hashimoto, and S. Kithata. 1990. Purification and some properties of ${\beta}$-furctofuranosidase I from Arthrobacter sp. K-1. Agric. Biol. Chem. 54: 913-919
  9. Hirayama, M., N. Sumi, and H. Hidaka. 1989. Purification and properties of a fructooligosaccharides-producing fructofuranosidase from Aspergillus niger ATCC 20611. Agric. Biol. Chem. 53: 667-673
  10. Houck, C. M., J. R. Pear, R. Elliott, and J. T. Perchorowicz. 1987. Isolation of DNA encoding sucrase genes from Streptococcus salivarius and partial characterization of the enzymes expressed in Escherichia coli. J. Bacteriol. 169: 3679-3684
  11. Kidby, D. K. and R. Davies. 1970. Invertase and disulphide bridges in the yeast wall. J. Gen. Microbiol. 61: 327-333
  12. Kim, B. M. 1980. Studies on invertase from Kroean ginseng, Panax ginseng C. A. Meyer. Korean J. Food Sci. Technol. 12: 1-5
  13. Krishnan, H. B., J. T. Blanchette, and T. W. Okita. 1985. Wheat invertase. Plant Physiol. 78: 241-245 https://doi.org/10.1104/pp.78.2.241
  14. Nakagawa, H., Y. Kawasaki, N. Ogura, and H. Takehata. 1971. Purification and some properties two types of ${\beta}$-fructofuranosidase from tomato fruit. Agric. Biol. Chem. 36: 18-26
  15. Obenland, D., U. Simmen, T. Boller, and A. Wiemken. 1993. Purification and characterization of three soluble invertase from barley leaves. Plant Physiol. 101: 1331-1339 https://doi.org/10.1104/pp.101.4.1331
  16. Porntaveewat, W., T. Takayanagi, and K. Yokotsuka. 1994. Purification and properties of invertase from Muscat Bailey A grapes. J. Ferment. Bioeng. 78: 288-292 https://doi.org/10.1016/0922-338X(94)90359-X
  17. Pressey, R. 1966. Separation and properties of potato invertase and invertase inhibitor. Arch. Biochem. Biophys. 113: 667-674 https://doi.org/10.1016/0003-9861(66)90246-3
  18. Unger, C., J. Hofsteenge, and A. Sturm. 1992. Purification and characterization of a soluble ${\beta}$-fructofuranosidase from Daucus carota. Eur. J. Biochem. 204: 915-921 https://doi.org/10.1111/j.1432-1033.1992.tb16712.x
  19. Yanase, H., H. Fukushi, N. Ueda, Y. Maeda, A. Toyoda, and K. Toromura. 1991. Cloning, Sequencing, and characterization of the intracelluar invertase gene from Zymomonas mobilis. Agric. Biol. Chem. 55: 1383-1390
  20. Tanzer, J. M., A. T. Brown, and M. F. Mclnerney. 1973. Identification, preliminary characterization, and evidence for regulation of invertase in Streptococcus mutants. J. Bacteriol. 116: 192-202
  21. Ottolenghi, P. 1971. A comparison of five genetically distinct invertase from Saccharomyces. Eur. J. Biochem. 18: 544-552 https://doi.org/10.1111/j.1432-1033.1971.tb01275.x
  22. Miller, G. L. 1959. Uses of dinitrosalicylic acid reagent for determination of reducing sugar. Anal. Chem. 31: 426-428 https://doi.org/10.1021/ac60147a030
  23. Rodriguez, R. L. and R. C. Tait. 1983 Recombinant DNA techniques: Introduction. pp. 37-51. Addison-Wesley Pub. co. Massachusette