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Zhang, W., Liu, C., Wang, G., Ma, Y., Zhang, K., Zou, S., and Zhang, M. (2012). "Comparison of the expression in Saccharomyces cerevisiae of endoglucanase II from Trichoderma reesei and endoglucanase I from Aspergillus aculeatus," BioRes. 7(3), 4031-4045.

Abstract

Two distinct expression cassettes were synthesized by overlapping PCR for expressing the endoglucanase I gene (egl1) from Aspergillus aculeatus and the endoglucanase II gene (egl2) from Trichoderma reesei in a Saccharomyces cerevisiae host. One contained the anchored sequence from the S. cerevisiae cwp2 gene, while the other did not. The low and high copy number plasmids YCplac33 and YEplac195 were used. The enzymatic activities and viscosity changes in the YP-CMC medium varied between the eight recombinant yeast strains produced, and the greatest values were obtained with the YE-TrEII’ strain, which had an activity of 347.7 U/g dry cell weight (DCW) and viscosity at 12 h of 4.7% of the initial control value, respectively; YE-TrEII’ was YEplac195-based and contained T. reesei egl2 and no Cwp2 sequence. Strains YC-AaEI and YC-TrEII showed the lowest enzyme activitiy (80.5 and 30.4 U/g DCW, respectively) and viscosity changes at 12 h (20.5 and 26.2% of the initial control viscosity, respectively), which were YCplac33-based and contained the Cwp2 sequence. The results showed that gene copy number was the most significant factor to influence the expression of endoglucanases in S. cerevisiae, and the existence of Cwp2 sequence led to decreased enzymatic level and viscosity-reducing performance, while it was shown not to realize efficient surface display of these two endoglucanases.
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