| 研究报告 |
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| Cloning, Expression and property analysis of Arabinosidase in Pichia pastoris |
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Abstract SMART-RACE was performed after Isolating the total RNA of Armillariella tabescens to amplify the full-length cDNA of arabinosidase（GenBank Accession No. AJ620046）. Bioinformatics analysis was used to analyze the code frame of arabinosidase, to predict its structure and function. Recombinant plasmid pPIC9-AF was constructed and then electroporated into methylotrophic yeast Pichia pastoris GS115. The secreted 6×His fusion protein was purified to analyze its enzymology property. This arabinosidase had high activity at 30℃-35℃ under acid condition, and was stable within wide range of pH and temperature. It maintained about 80% activity at the range of pH4.0-8.0 and 20℃-40℃，which was wider than many other cloned arabinosidase. So it was worthy to go step further to study this enzyme. And recombinant expression provided a chance of highly expressing arabinosidase.
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Received: 09 October 2005
Published: 25 April 2006
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