Construction of recombinant Saccharomyces cerevisiae strains containing endo-1,4-β-glucanase gene of Aspergillus niger into the chromosomal HO locus
Keywords:
cellobiohydrolase, endo-1, 4-â-glucanase, Saccharomyces cerevisiae, gene expression, enzymatic activity, glycosylationAbstract
The vector carrying a selective marker gene resistant to geneticin (G418), a constitutive promoter and terminator of yeast gene glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was created for integration of genetic material into the chromosomal HO locus. Through the use of a genomic integration vector, new yeast strains with efficient expression of endo-1,4-â-glucanase of the fungus Aspergillus niger were obtained. Approaches used in this study has been shown to be perspective in the development of industrial strains for the fermentation of cellulose-containing materials.References
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15 Quay D. H. X., Bakar F. D. A., Rabu A., Said M., Illias R. M., Mahadi N. M., Hassan O., and Murad A. M. A.
Overexpression, purification and characterization of the Aspergillus niger endoglucanase, EglA, in Pichia pastoris
//African Journal of Biotechnology. - 2011. -V. 10, ‡‚ 11. - P. 2101-2111.
16 Hasper A.A., Dekkers E., van Mil M., van de Vondervoort P.J. I., de Graaff L.H. EglC, a new endoglucanase from
Aspergillus niger with major activity towards xyloglucan // Appl. Environ. Microb. -2002. -V. 68, ‡‚ 4.-P.1556-1560.
17 Hong J., Wang Y., Kumagai H., Tamaki H. Construction of thermotolerant yeast expressing thermostable cellulase
genes // J. Biotechnol. -2007. .V.130. . P.114-123.
4 Lynd L.R., van Zyl W.H., McBride J. E., Laser M. Consolidated bioprocessing of cellulosic biomass: an update //
Current Opinion in Biotechnology. -2005. -V.16, ‡‚ 5. .P. 577.583.
5 Van Rensburg R., Van Zyl W.H., Pretorius I.S. Engineering yeast for efficient cellulose degradation // Yeast. -1998.-V.14. -P.67.76.
6 Fujita Y., Ito J., Ueda M., Fukuda H., Kondo H. Synergistic saccharification, and direct fermentation to ethanol, of
amorphous cellulose by use of an engineered yeast strain codisplaying three types of cellulolytic enzymes// Appl.
Environ. Microbiol. -2004. -V.70. .P.1207.1212.
7 Voth W.P., Richards J.D., Shaw J.M., Stillman D.J. Yeast vectors for integration at the HO locus // Nucleic Acids
Res. -2001. .V.29, ‡‚12.
8 Herskowitz I., Rine J., Strathern J. Mating type determination and mating-type interconversion in Saccharomyces
cerevisiae. In: The Molecular and Cellular Biology of the Yeast Saccharomyces: Gene Expression. Jones E.W., Pringle
J.R., Broach J.R. eds. Cold Spring Harbor Laboratory Press, Cold Spring Harbor - NYk, 1992. -P. 583-656.
9 Hong J., Tamaki H., Akiba S., Yamamoto K., Kumagai H. Cloning of a gene encoding a highly stable endo-b-1,4-ƒÀ-
glucanase from Aspergillus niger and its expression in yeast // J. Biosci. Bioeng. -2001. -V.92. -P. 434-441.
10 Miller, G.L. Use of dinitrosalicylic acid reagent for determination of reducing sugar//Anal. Chem.-1959. -Vol. 31.-P. 426.428.
11 Ornstein L., Davis B.J. Disc electrophoresis-I: Background and theory // Ann. NY. Acad. Sci. - 1964. - V.121. -P.321.349.
12 Laemmli U.K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4 //Nature. -1970.-V. 227.-P. 680-685.
13 Kasturi L., Chen H., Shakin-Eshleman S.H. Regulation of N-linked core glycosylation: use of a site-directed
mutagenesis approach to identify Asn-Xaa-Ser/Thr sequons that are poor oligosaccharide acceptors // Biochem. J. -1997. -V. 323. -P.415-419.
14 Maley F. et al. Characterization of glycoproteins and their associated oligosaccharides through the use of
endoglycosidases // Anal. Biochem. -1989. -V. 180. -P.195-204.
15 Quay D. H. X., Bakar F. D. A., Rabu A., Said M., Illias R. M., Mahadi N. M., Hassan O., and Murad A. M. A.
Overexpression, purification and characterization of the Aspergillus niger endoglucanase, EglA, in Pichia pastoris
//African Journal of Biotechnology. - 2011. -V. 10, ‡‚ 11. - P. 2101-2111.
16 Hasper A.A., Dekkers E., van Mil M., van de Vondervoort P.J. I., de Graaff L.H. EglC, a new endoglucanase from
Aspergillus niger with major activity towards xyloglucan // Appl. Environ. Microb. -2002. -V. 68, ‡‚ 4.-P.1556-1560.
17 Hong J., Wang Y., Kumagai H., Tamaki H. Construction of thermotolerant yeast expressing thermostable cellulase
genes // J. Biotechnol. -2007. .V.130. . P.114-123.
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