| Title |
Remodeling of the Glycosylation Pathway in the Methylotrophic Yeast Hansenula polymorpha to Produce Human Hybrid-Type N-Glycans |
| Author |
Seon Ah Cheon1,2, Hyunah Kim3, Doo-Byoung Oh2, Ohsuk Kwon2, and Hyun Ah Kang1,3* |
| Address |
1Research Center for Biomolecules and Biosystems, College of Natural Science, Chung-Ang University, Seoul 156-756, Republic of Korea, 2Systems and Synthetic Biology Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 305-806, Republic of Korea, 3Department of Life Science, College of Natural Science, Chung-Ang University, Seoul 156-756, Republic of Korea |
| Bibliography |
Journal of Microbiology, 50(2),341-348, 2012,
|
| DOI |
|
| Key Words |
Hansenula polymorpha, humanized hybrid-type N-glycans, ALG3, OCH1 |
| Abstract |
As a step forward to achieve the generation of human complex-
type N-glycans in the methylotrophic yeast Hansenula
polymorpha, we here report the modification of the yeast
glycosylation pathway by heterologous expression of the
human gene encoding β-1,2-N-acetylglucosaminyltransferase
I (GnTI). For the optimal expression of human GnTI in
the yeast Golgi compartment, the catalytic domain of the
GnTI was fused to various N-terminal leader sequences derived
from the yeast type II membrane proteins. The vectors
containing GnTI fusion constructs were introduced into
the H. polymorpha och1Δ single and och1Δalg3Δ double
mutant strains expressing the ER-targeted Aspergillus saitoi
α-1,2 mannosidase, respectively. Both of the glycoengineered
Hpoch1Δ and Hpoch1ΔHpalg3Δ strains were shown to produce
successfully the hybrid-type glycans with a monoantennary
N-acetylglucosamine (GlcNAc1Man5GlcNAc2 and
GlcNAc1Man3GlcNAc2, respectively) by N-glycan profile
analysis of cell wall proteins. Furthermore, by comparative
analysis of byproduct formation and the glycosylation site
occupancy, we propose that the Hpoch1Δ strain would be
more suitable than the Hpoch1ΔHpalg3Δ strain as a host
for the production of recombinant proteins with humanized
glycans. |
