Glycoengineering of the methylotrophic yeast Hansenula polymorpha for the production of glycoproteins with trimannosyl core N-glycan by blocking core oligosaccharide assembly.
- Authors
- Oh, Doo-Byoung; Park, Jeong-Seok; Kim, Moo Woong; Cheon, Seon Ah; Kim, Eun Jung; Moon, Hye Yun; Kwon, Ohsuk; Rhee, Sang Ki; Kang, Hyun Ah
- Issue Date
- May-2008
- Publisher
- Wiley - VCH Verlag GmbH & CO. KGaA
- Keywords
- ALG3; Glycoengineering; Hansenula polymorpha; Trimannosyl core N-glycan
- Citation
- Biotechnology Journal, v.3, no.5, pp 659 - 668
- Pages
- 10
- Journal Title
- Biotechnology Journal
- Volume
- 3
- Number
- 5
- Start Page
- 659
- End Page
- 668
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/57517
- DOI
- 10.1002/biot.200700252
- ISSN
- 1860-6768
1860-7314
- Abstract
- The initial lipid-linked oligosaccharide Glc3Man9GlcNAc2-dolichyl pyrophosphate (Dol-PP) for N-glycan is synthesized and assembled at the membrane of the endoplasmic reticulum (ER) and subsequently transferred to a nascent polypeptide by the oligosaccharide transferase complex. We have identified an ALG3 homolog (HpALG3) coding for a dolichyl-phosphate-mannose dependent α-1,3-mannosyltransferase in the methylotrophic yeast Hansenula polymorpha. The detailed analysis of glycan structure by linkage-specific mannosidase digestion showed that HpALG3 is responsible for the conversion of Man5GlcNAc2-Dol-PP to Man6GlcNAc2-Dol-PP, the first step to attach a mannose to the lipid-linked oligosaccharide in the ER. The N-glycosylation pathway of H. polymorpha has been remodeled by deleting the HpALG3 gene in the Hpoch1 null mutant strain blocked in the yeast-specific outer mannose chain synthesis and by introducing an ER-targeted Aspergillus saitoi α-1,2-mannosidase gene. This glycoengineered H. polymorpha strain produced glycoproteins mainly containing trimannosyl core N-glycan (Man3GlcNAc2), which is the common core backbone of various human-type N-glycans. The results demonstrate the high potential of H. polymorpha to be developed as an efficient expression system for the production of glycoproteins with humanized glycans.
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Collections - College of Natural Sciences > Department of Life Science > 1. Journal Articles
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