Applied Microbiology and Biotechnology

, Volume 103, Issue 7, pp 3085–3097 | Cite as

Identification and evaluation of novel anchoring proteins for cell surface display on Saccharomyces cerevisiae

  • Apisan Phienluphon
  • Wuttichai Mhuantong
  • Katewadee Boonyapakron
  • Pacharawan Deenarn
  • Verawat Champreda
  • Duangdao Wichadakul
  • Surisa SuwannarangseeEmail author
Applied genetics and molecular biotechnology


The development of arming yeast strains as whole-cell biocatalysts involves a selection of effective anchoring proteins to display enzymes and proteins on yeast cell surface. To screen for novel anchoring proteins with improved efficiency, a bioinformatics pipeline for the identification of glycosylphosphatidylinositol-anchored cell wall proteins (GPI-CWPs) suitable for attaching passenger proteins to the cell surface of Saccharomyces cerevisiae has been developed. Here, the C-terminal sequences (CTSs) of putative GPI-CWPs were selected based on the criteria that the sequence must contain a serine/threonine-rich (S/T) region of at least 30% S/T content, a total threonine content of at least 10%, a continuous S/T stretch of at least 130 amino acids in length, and a continuous T-rich region of at least 10 amino acids in length. Of the predicted 790 proteins, 37 putative GPI-CWPs were selected from different yeast and fungal species to be evaluated for their performance in displaying yeast-enhanced green fluorescent protein and β-glucosidase enzyme. This led to the identification of five novel anchoring proteins with higher performance compared to α-agglutinin used as benchmark. In particular, the CTS of uncharacterized protein in Kluyveromyces lactis, namely 6_Kl, is the most efficient anchoring protein of the group. The CTS of 6_Kl protein provided a β-glucosidase activity of up to 23.5 U/g cell dry weight, which is 2.8 times higher than that of the CTS of α-agglutinin. These identified CTSs could be potential novel anchoring protein candidates for construction of efficient arming yeasts for biotechnology applications in the future.


Cell surface display Anchoring protein GPI-anchored protein Saccharomyces cerevisiae β-Glucosidase Bioinformatics 



We thank Dr. Wananit Wimuttisuk and Mr. Christopher Keith Campbell for critically reading the manuscript.

Funding information

This work was financially supported by the National Center for Genetic Engineering and Biotechnology, Thailand (Grant no. P1300748).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with animals or human participants performed by any of the authors.

Supplementary material

253_2019_9667_MOESM1_ESM.pdf (285 kb)
ESM 1 (PDF 284 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Microbial Biotechnology and Biochemicals Research Unit, National Center for Genetic Engineering and Biotechnology (BIOTEC)National Science and Technology Development AgencyPathumthaniThailand
  2. 2.Department of Computer Engineering, Faculty of EngineeringChulalongkorn UniversityBangkokThailand

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