A cell surface display system using novel GPI-anchored proteins in Hansenula polymorpha

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Title
A cell surface display system using novel GPI-anchored proteins in Hansenula polymorpha
Author(s)
So-Young Kim; Jung Hoon Sohn; Y R Pyun; Eui Sung Choi
Bibliographic Citation
Yeast, vol. 19, no. 13, pp. 1153-1163
Publication Year
2002
Abstract
A cell surface display system was developed in yeast Hansenula polymorpha.The J four genes HpSED1, HpGAS1, HpTIP1 and HpCWP1, encoding glycosylphosphatidylinositol (GPI)-anchored cell surface proteins from H. polymorpha, were cloned, characterized and evaluated for their efficacies as cell surface display motifs of reporter proteins. Sequence analysis of these genes revealed that each encodes a typical GPI-anchored protein that is structurally similar to a counterpart gene in S. cerevisiae. The genes showed a high content of serine-threonine (alanine) and harboured a putative secretion signal in the N-terminus and the GPI-attachment signal in the C-terminus. The surface anchoring efficiency of these putative cell surface proteins was tested by fusion to the C-terminal of carboxymethy1cellulase (CMCase) from Bacillus subtilis. In all cases, high CMCase activities were detected in intact cell fraction, indicating anchoring of CMCase to the cell surface. HpCwp1p, HpGas1p and the 40 C-terminal amino acids of HpTip1p from H. polymorpha exhibited a comparatively high CMCase surface anchoring efficiency. When these proteins were used as anchoring motifs for surface display of the glucose oxidase (GOD) from Aspergillus niger, most enzyme activity was detected at the cell surface. Fluorescence activated cell sorter (FACS) analysis of cells displaying GOD on the cell surface demonstrated that GOD was well exposed on the cell surface. HpCwp1p showed the highest anchoring efficiency among others.
Keyword
CMCaseGlucose oxidaseGPI-anchored proteinHansenula polymorphaSurface display
ISSN
0749-503X
Publisher
Wiley
DOI
http://dx.doi.org/10.1002/yea.911
Type
Article
Appears in Collections:
Synthetic Biology and Bioengineering Research Institute > Synthetic Biology Research Center > 1. Journal Articles
Division of Bio Technology Innovation > BioProcess Engineering Center > 1. Journal Articles
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