Cell-Surface Expression of Aspergillus saitoi-Derived Functional α-1,2-Mannosidase on Yarrowia lipolytica for Glycan Remodeling

Hye Yun Moon; Trinh Luu Van; Seon Ah Cheon; Jinho Choo; Jeong-Yoon Kim; Hyun Ah Kang

doi:10.1007/s12275-013-3344-x

Articles

Page Path: HOME > J. Microbiol > Volume 51(4); 2013 > Article

Research Support, Non-U.S. Gov't
Cell-Surface Expression of Aspergillus saitoi-Derived Functional α-1,2-Mannosidase on Yarrowia lipolytica for Glycan Remodeling: Hye Yun Moon ¹, Trinh Luu Van ¹, Seon Ah Cheon ¹, Jinho Choo ¹, Jeong-Yoon Kim ², Hyun Ah Kang ¹; Journal of Microbiology 2013;51(4):506-514.
DOI: https://doi.org/10.1007/s12275-013-3344-x
Published online: August 30, 2013

¹Department of Life Science, Chung-Ang University, Seoul 156-756, Republic of Korea, ²Department of Microbiology, School of Bioscience and Biotechnology, Chungnam National University, Daejeon 305-764, Republic of Korea¹Department of Life Science, Chung-Ang University, Seoul 156-756, Republic of Korea, ²Department of Microbiology, School of Bioscience and Biotechnology, Chungnam National University, Daejeon 305-764, Republic of Korea

Corresponding author: Hyun Ah Kang , Tel: +82-2-820-5863,

Received: 26 June 2013 • Accepted: 17 July 2013

prev next

47 Views
0 Download
13 Crossref
16 Scopus

Full Article

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 biocatalysts. The hemiascomycetes yeast Yarrowia lipolytica has been raised as a potential host for heterologous expression of recombinant proteins. In this study, we report the expression of Aspergillus saitoi α-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 α-amylase and rice α-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 α-1,2-mannosidase activity assay using intact Y. lipolytica cells as the enzyme source. Furthermore, the glycoengineered Y. lipolytica Δoch1Δmpo1 strains displaying α-1,2-mannosidase were able to convert Man8GlcNAc2 to Man5GlcNAc2 efficiently on their cell-wall mannoproteins, demonstrating its potential used for glycoengineering in vitro or in vivo.

Keywords: Yarrowia lipolytica;surface display;a-1;2-mannosidase;GPI-anchor

Figure & Data

References

Citations

Citations to this article as recorded by

Engineering novel Yarrowia lipolytica whole-cell biocatalysts by cell surface display of the native Lip2 lipase for biodiesel production
Maria Orfanidou, Eleftheria Panagiotidou, Antonios M. Makris, Eleni Theodosiou
Biotechnology for the Environment.2025;[Epub] CrossRef
Yeast Surface Display System: Strategies for Improvement and Biotechnological Applications
Karla V. Teymennet-Ramírez, Fernando Martínez-Morales, María R. Trejo-Hernández
Frontiers in Bioengineering and Biotechnology.2022;[Epub] CrossRef
Dietary intake of GDF11 delays the onset of several biomarkers of aging in male mice through anti-oxidant system via Smad2/3 pathway
Lili Song, Fei Wu, Congjun Li, Shicui Zhang
Biogerontology.2022; 23(3): 341. CrossRef
Hydrolytic secretome engineering in Yarrowia lipolytica for consolidated bioprocessing on polysaccharide resources: review on starch, cellulose, xylan, and inulin
Ewelina Celińska, Jean-Marc Nicaud, Wojciech Białas
Applied Microbiology and Biotechnology.2021; 105(3): 975. CrossRef
Yarrowia lipolytica Strains and Their Biotechnological Applications: How Natural Biodiversity and Metabolic Engineering Could Contribute to Cell Factories Improvement
Catherine Madzak
Journal of Fungi.2021; 7(7): 548. CrossRef
Engineering Yarrowia lipolytica for Use in Biotechnological Applications: A Review of Major Achievements and Recent Innovations
Catherine Madzak
Molecular Biotechnology.2018; 60(8): 621. CrossRef
Synthetic biology tools for engineering Yarrowia lipolytica
M. Larroude, T. Rossignol, J.-M. Nicaud, R. Ledesma-Amaro
Biotechnology Advances.2018; 36(8): 2150. CrossRef
Development of recombinant Yarrowia lipolytica producing virus-like particles of a fish nervous necrosis virus
Van-Trinh Luu, Hye Yun Moon, Jee Youn Hwang, Bo-Kyu Kang, Hyun Ah Kang
Journal of Microbiology.2017; 55(8): 655. CrossRef
Using a vector pool containing variable-strength promoters to optimize protein production in Yarrowia lipolytica
Rémi Dulermo, François Brunel, Thierry Dulermo, Rodrigo Ledesma-Amaro, Jérémy Vion, Marion Trassaert, Stéphane Thomas, Jean-Marc Nicaud, Christophe Leplat
Microbial Cell Factories.2017;[Epub] CrossRef
Yarrowia lipolytica: recent achievements in heterologous protein expression and pathway engineering
Catherine Madzak
Applied Microbiology and Biotechnology.2015; 99(11): 4559. CrossRef
Biotechnological applications of Yarrowia lipolytica: Past, present and future
Hu-Hu Liu, Xiao-Jun Ji, He Huang
Biotechnology Advances.2015; 33(8): 1522. CrossRef
Functional characterization of extracellular chitinase encoded by the YlCTS1 gene in a dimorphic yeast Yarrowia lipolytica
Jeong-Nam Park, Chang Pyo Han, Dong-Jik Lee, Seon Ah Cheon, Hyun Ah Kang
Journal of Microbiology.2014; 52(4): 284. CrossRef
Characterization of putative glycosylphosphatidylinositol-anchoring motifs for surface display in the methylotrophic yeast Hansenula polymorpha
Seon Ah Cheon, Jinhee Jung, Jin Ho Choo, Doo-Byoung Oh, Hyun Ah Kang
Biotechnology Letters.2014; 36(10): 2085. CrossRef

Cite this Article

Cite this Article: export Copy Download Format; Close

Download Citation

Download a citation file in RIS format that can be imported by all major citation management software, including EndNote, ProCite, RefWorks, and Reference Manager.

Format:

RIS — For EndNote, ProCite, RefWorks, and most other reference management software
BibTeX — For JabRef, BibDesk, and other BibTeX-specific software

Include:

Citation for the content below
Citation and abstract for the content below

Cell-Surface Expression of Aspergillus saitoi-Derived Functional α-1,2-Mannosidase on Yarrowia lipolytica for Glycan Remodeling

J. Microbiol. 2013;51(4):506-514. Published online August 30, 2013

DOI: https://doi.org/10.1007/s12275-013-3344-x

Related articles