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| The Stability Reconstruction of β-mannanase with N-glycosylation Modification |
| XIE Chun-fang1,3, LI Yu-feng2,3, LIU Da-ling1,3, YAO Dong-sheng2,3,4 |
1. Department of Bioengineering, Jinan University, Guangzhou 510632, China;
2. Institute of Biomedicine, Jinan University, Guangzhou 510632, China;
3. Guangdong Provincial Key Laboratory of Bioengineering Medicine, Guangzhou 510632, China;
4. National Engineering Research Center of Genetic Medicine, Guangzhou 510632, China |
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Abstract N-glycosylation is one of the most important posttranslational modification of proteins in eukaryotes. A rational strategy to introduce N-glycosylation site was proposed to Armillariella tabescens beta mannose Man47.Then g-123 mutant with EAS(enhanced aromatic sequence) sequence was built through the molecular docking, secondary structure analysis and feasibility analysis of glycosylation. The sequence of g-123 mutant was inserted into SMD1168 with the yeast α-mating factor, then transformed it into Pichia by electroporation to obtain the recombinants. Finally the thermal stability, acid and alkali stability, pepsin-resistance and trypsin-resistance of g-123 and wild type were analyzed. The results showed that compared with wild type, the thermal stability, acid and alkali stability, protease resistance of the mutant g-123 with glycosylation was improved.
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Received: 03 September 2013
Published: 25 December 2013
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