Browse > Article

Expression of Tkermomonoepora fusea Exoglucanase in Saccharomyces cerevisiae and Its Application to Cellulose Hydrolysis  

Park Hyun-Soon (Department of Biotechnology & Bioengineering, Pukyung National University)
Kim Hyun-Chul (Biomaterial Control, Dong-Eui University)
Shin Dong-Ha (Insect Biotech Co.)
Kim Joong-Kyun (Department of Biotechnology & Bioengineering, Pukyung National University)
Nam Soo-Wan (Department of Biotechnology & Bioengineering, Dong-Eui University)
Publication Information
Microbiology and Biotechnology Letters / v.33, no.4, 2005 , pp. 267-273 More about this Journal
Abstract
To develop effective and powerful probiotic, Saccharomyces cerevisiae strains producing cellulolytic enzymes were genetically brooded. For the production of exoglucanase, the plasmid pVT-TExo (8.8 kb) was constructed, in which Thermomonosporafusca exoglucanase gene (E3) was under the control of ADHl promoter, and introduced into S. cerevisiae SEY2102. When the transformant, S. cerevisiae SEY2102/pVT-TExo, was cultivated on YPD medium, the total expression level of avicelase reached about 190 unit/l. The secretion efficiency and plasmid stability were about $50\%\;and\;91\%$, respectively. Recombination exoglucanase enzyme bound to avicel better than Clostridium endoglucanase (CelA) and Trichoderma endoglucanase (C4) enzymes. The mixing ratio of E3 and CelA displaying the best synergistic hydrolysis for avicel was observed at 4:1. The mixture of endoglucanase (CelA) and exoglucanase (E3) resulted in 3.2-fold increase of avicelase activity and 2.5-fold enhanced production of sugar production from avicel, compared to the single enzyme treatment.
Keywords
Saccharomyces cerevisiae; exoglucanase; endoglucanase synergistic hydrolysis; cellulose;
Citations & Related Records

Times Cited By SCOPUS : 0
연도 인용수 순위
  • Reference
1 Draborg, H., S. Christgau, T. Halkier, G. Rasmussen, H. Dalboge, and S. Kauppinen. 1996. Secretion of an enzymatically active Trichoderma harzianum endochitinase by Saccharomyces cerevisiae. Curr. Genet. 29: 404-409   DOI   ScienceOn
2 Kim, D. W., Y. H. Jang, and Y. K. Jeong, 1997. Adsorption behaviours of two cellobiohydrolase and thier core proteins from Trichoderma reesei on avicel PH101. Biotechnol. Lett. 19: 893-897   DOI   ScienceOn
3 Maras, M., A. De Bruyn, J. Schraml, P. Herdewijn, M. Claeyssens, W. Fiers, and R. Contreras. 1997. Structural characterization of N-linked oligosaccharides from cellobiohydrolase I secreted by the filamentous fungus Trichoderma reesei RUTC 30. Eur. J. Biochem. 245: 617-625   DOI   ScienceOn
4 Suvajittanont, W., J. McGuire, and M. K. Bothwell. 2001. Adsorption of Thermomonospora fusca E(5) cellulase on silanized silica. Biotechnol. Bioeng. 67: 12-18
5 Teeri, T. T., P. Lehtovaara, S. Kauppinen, I. Salovuori, and J. K. C. Knowles. 1983. Homologous domains in Trichoderma reesei cellulolytic enzymes: gene sequence and expression of cellobiohydrolase II. Gene 51: 43-52   DOI   ScienceOn
6 Wong, W. K. R., C. Curry., R. S. Parekh., S. R. Parekh., M. Wayman., R. W. Davies., D. G Kilburn, and N. Skipper. 1988. Wood hydrolysis by Cellulomonas fimi endoglucanase and exoglucanase coexpressed as secreted enzymes in Saccharomyces cerevisiae. Bio/Technol. 6: 713-719   DOI
7 Zhang, S., G Lau, and D. B. Wilson. 1995. Characterization of Thermomonospora fusca exoglucanase. Biochemistry 34: 3386-3395   DOI   ScienceOn
8 Correa, M. G. and R. P. Tengerd. 1991. Production of cellulase on sugar cane bagasse by fungal mixed culture solid substrate fermentation. Biotechnol. Left. 19: 665-667
9 Barnett, C. C., R. M. Berka, and T. Fowler. 1991. Cloning and amplification of the gene encoding an extracellular ${\beta}$-glucosidase from Trichoderma reesei: evidence for improved rates of saccharification of cellulosic substrates. Bio/Technol. 9: 562-567   DOI   ScienceOn
10 Nam, S. W., D. K. Chung, and B. H. Chung. 1997. Constitutive expression of Clostridium thermocellum endoglucanase gene in Saccharomyces cerevisiae. Kor. J. Biotechnol. Bioeng. 12: 430-437
11 Vemet, T., D. Dignard, and D. Y. Thomas. 1987. A family of yeast expression vectors containing the phage f1 intergenic region. Gene 52: 225-233   DOI   PUBMED   ScienceOn
12 Mach R. L. and S. Zeilinger. 2003. Regulation of gene expression in industrial fungi, Trichoderma. Appl. Microbiol. Biotechnol. 60: 515-522   DOI
13 Son, M. I. and J. O. Kim, 1998 Optimum condition of cellulose hydrolysis reaction with mixed enzymes of cellulase and ${\beta}$-glucosidase. Kor. J. Biotechnol. Bioeng. 13: 20-25
14 Shin, D. H., J. B. Kim, B. W. Kim, and S. W. Nam. 1998. Expression and secretion of Trichoderma endoglucanase in Saccharomyces cerevisiae. Kor. J. Appl. Microbiol. Biotechnol. 26: 406-412
15 Irwin, D. C., M. Speaiw, L. P. Walker, and D. B. Wilson. 1993. Activity studies of eight purified cellulase: specificity, synergism, and binding effect. Biotechnol. Bioeng. 42: 1002-1013   DOI   ScienceOn
16 Kruus, K. and A. Andreacchi. 1995. Product inhibition of the recombinant CeIS, an exoglucanae component of the Clostrdium thermocellum cellulosome. Appl. Microbiol. Biotechnol. 44: 399-404   DOI   ScienceOn
17 Wang, T., L. Xiangmei, Y. Qian, Z. Xi, Q. Yinbo, G Peiji, and W. Tianhong. 2005. Directed evolution for engineering pH profile of endoglucanase III from Trichoderma reesei. Biomol. Eng. 22: 89-94   DOI   ScienceOn
18 Nam, S. W., K. Yoda, and M. Yamasaki. 1993. Secretion and localization of invertase and inulinase in recombinant Saccharomyces cerevisiae. Biotechnol. Lett. 15: 1049-1504   DOI   ScienceOn
19 Hoshino, E., M. Shiroshi, Y. Amano, M. Nomura, and T. Kanda, 1997. Synergistic action of exo-type cellulase in the hydrolysis of cellulase with different crystallinities. J. Ferment. Bioeng. 84: 300-306   DOI   ScienceOn
20 Medve, J. J. Karlsson, D. Lee, and F. Tjerneld. 1998. Hydrolysis of microcrystalline cellulose by cellobiohydrolae I and endoglucanase II from Trichoderma reesei: adsorption, sugar production pattern, and synergism of the enzyme. Biotechnol. Bioeng. 59: 621-634   DOI   ScienceOn
21 Penttila, M., L. Ander, M. Saloheimo, P. Lehtovaara, and J. K. C. Knowles. 1987. Expression of two Trichoderma reesei endoglucanase in yeast Saccharomyces cerevisiae. Yeast 3: 175-185   DOI   ScienceOn
22 Ito, H., Y. Fukuda, K. Murata, and A. Kimura. 1983. Transformation of intact yeast cells treated with alkali cations. J. Bacteriol. 153: 163-168   PUBMED
23 Skiper, N., M. Sutherland, R. W. Davies, D. Kilburn, R. C. Miller, Jr., A. Warren, and R. Womg. 1985. Secretion of a bacterial cellulase by yeast. Science 230: 958-960   DOI   PUBMED   ScienceOn
24 Esterbauer, H., W. Steiner, I. Labudova, A. Hermann, and M. Hayn. 1991. Production of Trichoderma cellulase in laboratory and pilot plant. Biores. Technol. 36: 67-76   DOI   ScienceOn
25 Wood, T. M. and S. J. Mccrae. 1978. The cellulase Trichoderma koningii: purification and properties of some endoglucanase components with special reference to their action on cellulose when action along in synergism with the cellobiohydrolase. J. Biochem. 171: 61-72   DOI
26 Miller, G. L., R. Blum, W. E. Glennon, and A. L. Burton. 1960. Measurement of carboxymethyl cellulase activity. Anal. Biochem. 2: 127-132