Cell-Surface Expression of Aspergillus saitoi-Derived Functional alpha-1,2-Mannosidase on Yarrowia lipolytica for Glycan Remodelingopen access
- Authors
- Moon, Hye Yun; Van, Trinh Luu; Cheon, Seon Ah; Choo, Jinho; Kim, Jeong-Yoon; Kang, Hyun Ah
- Issue Date
- Aug-2013
- Publisher
- MICROBIOLOGICAL SOCIETY KOREA
- Keywords
- Yarrowia lipolytica; surface display; a-1,2-mannosidase; GPI-anchor
- Citation
- JOURNAL OF MICROBIOLOGY, v.51, no.4, pp 506 - 514
- Pages
- 9
- Journal Title
- JOURNAL OF MICROBIOLOGY
- Volume
- 51
- Number
- 4
- Start Page
- 506
- End Page
- 514
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/14434
- DOI
- 10.1007/s12275-013-3344-x
- ISSN
- 1225-8873
1976-3794
- Abstract
- Expression of proteins on the surface of yeast has a wide range of applications, such as development of live vaccines, screening of antibody libraries, and use as whole-cell bio-catalysts. The hemiascomycetes yeast Yarrowia hpolytica has been raised as a potential host for heterologous expression of recombinant proteins. In this study, we report the expression of Aspergillus saitoi alpha-1,2-mannosidase, encoded by the msdS gene, on the cell surface of Y. lipolytica. As the first step to achieve the secretory expression of msdS protein, four different signal sequences-derived from the endogenous Y. lipolytica Lip2 and Xpr2 prepro regions and the heterologous A. niger alpha-amylase and rice a-amylase signal sequences-were analyzed for their secretion efficiency. It was shown that the YlLip2 prepro sequence was most efficient in directing the secretory expression of msdS in fully N-glycosylated forms. The surface display of msdS was subsequently directed by fusing GPI anchoring motifs derived from Y. lipolytica cell wall proteins, YlCwp1p and YlYwp1p, respectively, to the C-terminus of the Lip2 prepro-msdS protein. The expression of actively functional msdS protein on the cell surface was confirmed by western blot, flow cytometry analysis, along with the alpha-1,2-mannosidase activity assay using intact Y. lipolytica cells as the enzyme source. Furthermore, the glycoengineered Y. lipolytica Delta och1 Delta mpo1 strains displaying alpha-1,2-mannosidase were able to convert Man(8)GlcNAc(2) to Man(5)GlcNAc(2) efficiently on their cell-wall mannoproteins, demonstrating its potential used for glycoengineering in vitro or in vivo.
- Files in This Item
-
- Appears in
Collections - College of Natural Sciences > Department of Life Science > 1. Journal Articles
![qrcode](https://api.qrserver.com/v1/create-qr-code/?size=55x55&data=https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/14434)
Items in ScholarWorks are protected by copyright, with all rights reserved, unless otherwise indicated.