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Cloning and expression of N-glycosylation-related mannosidase from Glaciozyma antarctica for the production of a mannosynthase

Elangovan, Dharshini; Kamaruddin, Shazilah; Hashim, Noor Haza Fazlin; Bakar, Farah Diba Abu; Murad, Abd. Munir Abd.; Mahadi, Nor Muhammad ; Allman, Sarah Ann and Mackeen, Mukram Mohamed (2016). Cloning and expression of N-glycosylation-related mannosidase from Glaciozyma antarctica for the production of a mannosynthase. In: AIP Conference Proceedings, AIP Publishing, Melville, NY, 1784, article no. 30011.

DOI (Digital Object Identifier) Link: https://doi.org/10.1063/1.4966749
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Abstract

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.

Item Type: Conference or Workshop Item
Copyright Holders: 2016 The Authors
ISBN: 0-7354-1446-7, 978-0-7354-1446-4
ISSN: 1551-7616
Extra Information: Proceedings of the Universiti Kebangsaan Malaysia, Faculty of Science and Technology 2016 Postgraduate Colloquium
Academic Unit/School: Faculty of Science, Technology, Engineering and Mathematics (STEM) > Life, Health and Chemical Sciences
Faculty of Science, Technology, Engineering and Mathematics (STEM)
Interdisciplinary Research Centre: Biomedical Research Network (BRN)
Item ID: 47959
Depositing User: Sarah Allman
Date Deposited: 12 Dec 2016 09:19
Last Modified: 12 Dec 2016 09:19
URI: http://oro.open.ac.uk/id/eprint/47959
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