Novel Properties for Endoglucanase Acquired by Cell-Surface Display Technique

In order to improve the stability of endoglucanase under thermal and acidic conditions, the endoglucanase gene was fused to the N-terminus of the Saccharomyces cerevisiae pir gene, encoding the cell wall protein PIR. The fusion gene was transformed into Pichia pastoris GS115 for expression. A result...

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Veröffentlicht in:Journal of microbiology and biotechnology 2015, Vol.25 (11), p.1856-1862
Hauptverfasser: Shi, Baosheng, Ke, Xiaojing, Yu, Hongwei, Xie, Jing, Jia, Yingmin, Guo, Runfang
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container_issue 11
container_start_page 1856
container_title Journal of microbiology and biotechnology
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creator Shi, Baosheng
Ke, Xiaojing
Yu, Hongwei
Xie, Jing
Jia, Yingmin
Guo, Runfang
description In order to improve the stability of endoglucanase under thermal and acidic conditions, the endoglucanase gene was fused to the N-terminus of the Saccharomyces cerevisiae pir gene, encoding the cell wall protein PIR. The fusion gene was transformed into Pichia pastoris GS115 for expression. A resulting strain with high expression and high activity was identified by examining resistance to Geneticin 418, Congo red staining, and quantitative analysis of enzyme activity. SDS-PAGE analysis revealed that the endoglucanase was successfully displayed on the yeast cell surface. The displayed endoglucanase (DEG) showed maximum activity towards sodium carboxyl methyl cellulose at approximately 275 IU/g cell dry weight. DEG exhibited greater than 60% residual activity in the pH range 2.5-8.5, higher than free endoglucanase (FEG), which had 40% residual activity at the same pH range. The highest tolerated temperature for DEG was 70℃, much higher than that of FEG, which was approximately 50℃. Moreover, DEG showed 91.1% activity at 65℃ for 120 min, while FEG only kept 77.8% residual activity over the same period. The half-life of DEG was 270 min at 65℃, compared with only 150 min for FEG. DEG could be used repeatedly at least three times. These results suggest that the DEG has broad applications as a yeast whole-cell biocatalyst, due to its novel properties of high catalytic efficiency, acid-thermal stabilities, and reusability.
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title Novel Properties for Endoglucanase Acquired by Cell-Surface Display Technique
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