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

• Published in: AIP Conference Proceedings,
• Main Authors: Elangovan, Dharshini, Kamaruddin, Shazilah, Hashim, Noor Haza Fazlin, Bakar, Farah Diba Abu, Murad, Abd. Munir Abd., Mahadi, Nor Muhammad, Allman, Sarah Ann, Mackeen, Mukram Mohamed
• Format: Conference Object
• Language: unknown
• Published: AIP Publishing 2016
• Subjects: Antarc*; Antarctica; antartic*
• Online Access: https://oro.open.ac.uk/47959/; https://doi.org/10.1063/1.4966749

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.