CHARACTERISTICS COMPARISON BETWEEN PLASMID-HARBORING AND CHROMOSOME-INTEGRATED RECOMBINANT SACCHAROMYCES CEREVISIAE CULTURES

Cha HJ; Yoo YJ

Korean Journal of Chemical Engineering, Vol.12, No.5, 567-571, November, 1995

Cha HJ, Yoo YJ

It is important to understand the differences in characteristics between plasmid-harboring recombinant yeast and chromosome-integrated recombinant yeast for the design and optimization of bioprocess employing recombi-nant yeast. In the present study, heterologous glucoamylase gene was inserted into yeast. The glucoamylase activity per gene copy number of chromosome-integrated recombinant yeast MMY2SUCSTA-I-5 was 3 to 6 folds higher than that of plasmid-harboring recombinant yeast MMY2SUCSTA. And the genetic stability of chromosome-integrated recombinant yeast (99%) was far better compared to plasmid-harboring recombinant yeast (65%). Better genetic stability and glucoamylase activity per gene copy number of chromosome-integrated recombinant yeast can provide advantages in higher final expression level, especially in continuous culture, compared to the plasmid-harboring recombinant yeast. The optimal glucose concentration for maximum expression of glucoamylase in chromosome-integrated recombinant yeast was lower than that in plasmid-harboring recombinant yeast.

Keywords:Saccharomyces cerevisiae;Plasmid-harboring Recombinant Yeast;Chromosome-integrated Recombinant Yeast;Glucoamylase;Culture Characteristics

[References]

Ahn JH, "Regulation of Glucoamylase Gene Expression in Saccharomyces cerevisiae var. diastaticus,", Ph.D. Thesis, Seoul National Univ., Seoul, Korea (1994)
Barbotin JN, Sayadi S, Nasri M, Berry F, Thomas D, "Improvement of Plasmid Stability by Immobilization of Recombinant Microorganisms," Annals of the New york Academy of Science, 589, 41 (1990)
Birnboim HC, Doly J, Nucl. Acids Res., 7, 1513 (1979)
Blomqvist K, Suihko ML, Knoeles J, Penttila M, Appl. Environ. Microbiol., 57, 2796 (1991)
Carlson M, Botstein D, Cell, 28, 145 (1982)
Cha HJ, Yoo YJ, Ahn JH, Kang HS, Biotechnol. Lett., 14, 747 (1992)
Chu FK, Maley F, J. Biol. Chem., 255, 6392 (1980)
DaSilva NA, Bailey JE, Biotechnol. Bioeng., 37, 318 (1991)
Fogel S, Welch JW, Proc. Natl. Acad. Sci. USA, 79, 5342 (1982)
Hinnen A, Hicks JB, Fink GR, Proc. Natl. Acad. Sci. USA, 75, 1929 (1978)
Ito H, Fukuda Y, Murata K, Kimura A, J. Bacteriol., 153, 163 (1983)
Kingsman SM, Kingsman AJ, Mellor J, Trends Biotechnol., 5, 53 (1987)
Kushner SR, "An Improved Method with ColE1-Derived Plasmids," in Genetic Engineering (Eds. H.B. Boyer and S. Nicosia), Elsevier/North-Holland, Amsterdam, p. 17 (1978)
Marten MR, Seo JH, Biotechnol. Bioeng., 34, 1133 (1989)
Meacock PA, Cohan SN, Cell, 20, 529 (1980)
Rose MD, Methods Enzymol., 152, 481 (1987)
Sakai A, Shimizu Y, Hishinuma F, Appl. Microbiol. Biotechnol., 33, 302 (1990)
Seo JH, Bailey JE, Biotechnol. Bioeng., 27, 1668 (1985)
Siegel R, Ryu DY, Biotechnol. Bioeng., 27, 28 (1985)
Stockhaus J, Eckes P, Blau A, Schell J, Willmitzer L, Nucl. Acids Res., 15, 3479 (1987)
Summers DK, Sherratt DJ, Cell, 36, 1097 (1984)
Yamashita I, Suzuki K, Fukui S, J. Bacteriol., 161, 567 (1985)

[Referenced By]

[1] Ahn JH, "Regulation of Glucoamylase Gene Expression in Saccharomyces cerevisiae var. diastaticus,", Ph.D. Thesis, Seoul National Univ., Seoul, Korea (1994)

[2] Barbotin JN, Sayadi S, Nasri M, Berry F, Thomas D, "Improvement of Plasmid Stability by Immobilization of Recombinant Microorganisms," Annals of the New york Academy of Science, 589, 41 (1990)

[3] Birnboim HC, Doly J, Nucl. Acids Res., 7, 1513 (1979)

[4] Blomqvist K, Suihko ML, Knoeles J, Penttila M, Appl. Environ. Microbiol., 57, 2796 (1991)

[5] Carlson M, Botstein D, Cell, 28, 145 (1982)

[6] Cha HJ, Yoo YJ, Ahn JH, Kang HS, Biotechnol. Lett., 14, 747 (1992)

[7] Chu FK, Maley F, J. Biol. Chem., 255, 6392 (1980)

[8] DaSilva NA, Bailey JE, Biotechnol. Bioeng., 37, 318 (1991)

[9] Fogel S, Welch JW, Proc. Natl. Acad. Sci. USA, 79, 5342 (1982)

[10] Hinnen A, Hicks JB, Fink GR, Proc. Natl. Acad. Sci. USA, 75, 1929 (1978)

[11] Ito H, Fukuda Y, Murata K, Kimura A, J. Bacteriol., 153, 163 (1983)

[12] Kingsman SM, Kingsman AJ, Mellor J, Trends Biotechnol., 5, 53 (1987)

[13] Kushner SR, "An Improved Method with ColE1-Derived Plasmids," in Genetic Engineering (Eds. H.B. Boyer and S. Nicosia), Elsevier/North-Holland, Amsterdam, p. 17 (1978)

[14] Marten MR, Seo JH, Biotechnol. Bioeng., 34, 1133 (1989)

[15] Meacock PA, Cohan SN, Cell, 20, 529 (1980)

[16] Rose MD, Methods Enzymol., 152, 481 (1987)

[17] Sakai A, Shimizu Y, Hishinuma F, Appl. Microbiol. Biotechnol., 33, 302 (1990)

[18] Seo JH, Bailey JE, Biotechnol. Bioeng., 27, 1668 (1985)

[19] Siegel R, Ryu DY, Biotechnol. Bioeng., 27, 28 (1985)

[20] Stockhaus J, Eckes P, Blau A, Schell J, Willmitzer L, Nucl. Acids Res., 15, 3479 (1987)

[21] Summers DK, Sherratt DJ, Cell, 36, 1097 (1984)

[22] Yamashita I, Suzuki K, Fukui S, J. Bacteriol., 161, 567 (1985)