Biotechnology Letters

, Volume 32, Issue 8, pp 1131–1136

Surface display of active lipase in Pichia pastoris using Sed1 as an anchor protein

Original Research Paper

DOI: 10.1007/s10529-010-0270-4

Cite this article as:
Su, G., Zhang, X. & Lin, Y. Biotechnol Lett (2010) 32: 1131. doi:10.1007/s10529-010-0270-4

Abstract

A Pichia pastoris cell-surface display system was constructed using the Sed1 anchor system that has been developed in Saccharomyces cerevisiae. Candida antarctica lipase B (CALB) was used as the model protein and was fused to an anchor that consisted of 338 amino acids of Sed1. The resulting fusion protein CALBSed1 was expressed under the control of the alcohol oxidase 1 promoter (pAOX1). Immunofluorescence microscopy of immunolabeled Pichiapastoris revealed that CALB was displayed on the cell surface. Western blot analysis showed that the fusion protein CALBSed1 was attached covalently to the cell wall and was highly glycosylated. The hydrolytic activity of the displayed CALB was more than 220 U/g dry cells after 120 h of culture. The displayed protein also exhibited a higher degree of thermostability than free CALB.

Keywords

Candida antarctica lipase BImmunofluorescence microscopyLipasePichia pastorisYeast surface display

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  1. 1.Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Bioscience and BioengineeringSouth China University of TechnologyGuangzhouPeople’s Republic of China