Cloning and expression of N-glycosylation-related mannosidase from Glaciozyma antarctica for the production of a mannosynthase
The controlled synthesis of oligosaccharides is of growing interest due to the important roles of oligosaccharides in various biological processes. Enzymatic synthesis enables regio- and stereo-selective control during synthesis which still remains a challenge using total chemical synthesis. In this...
Published in: | AIP Conference Proceedings, |
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Main Authors: | , , , , , , , |
Format: | Conference Object |
Language: | unknown |
Published: |
AIP Publishing
2016
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Subjects: | |
Online Access: | https://oro.open.ac.uk/47959/ https://doi.org/10.1063/1.4966749 |
Summary: | The controlled synthesis of oligosaccharides is of growing interest due to the important roles of oligosaccharides in various biological processes. Enzymatic synthesis enables regio- and stereo-selective control during synthesis which still remains a challenge using total chemical synthesis. In this study, endoplasmic reticulum 1,2-α-mannosidase from Glaciozyma antractica was recombinantly expressed in Pichia pastoris . The gene sequence for ER mannosidase was obtained from the Glaciozyma antractica database. The BLAST (Basic Local Alignment Search Tool) results from bioinformatics screening showed that ER mannosidase had 41 % identity with the equivalent mannosidases from Sacchromyces cerevesiae . ER mannosidase from G. antartica was then cloned into the pPICZαC expression vector and used to transform in the host Pichia pastoris X33 cells. The ER mannosidase (MW∼58 kDa) was successfully expressed at 25 °C with 1.0 % methanol induction. |
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