Constructon of Yeast Surface Display of ProROL by Using Flo1p Anchor System

Abstract:

A Pichia pastoris cell-surface display system was constructed using a Flo1p anchor system containing N-terminal flocculation functional domain (874 residues, FS), derived from Saccharomyces cerevisiae. The lipase from Rhizopus oryzae with a pro sequence (ProROL) and green fluorescent protein (GFP) gene were successfully cloned and genetically fused to the anchor system with their C-terminus free. Fluorescence microscope was used to detect the GFP displayed on the recombinant P. pastoris cell surface. The results showed that the yeast surface display system using Flo1p as the anchor protein was successfully constructed, and the activity of ProROL displayed on KM71H reached to 217.15 U/g, much higher than 61.30 U/g reported by Matsumoto. Besides, the yeast surface display system could effectively shorten the fermentation time compared with traditional fermentation.

• Periodical: Advanced Materials Research (Volumes 512-515)
• Pages: 356-360
• DOI: https://doi.org/10.4028/www.scientific.net/AMR.512-515.356
• Online since: May 2012
• Authors: Ye Wang, Wen Qian Li, Xun Li, Hao Shi, Fei Wang
• Keywords: Anchor Protein, Flo1p, Pichia pastoris, Rhizopus oryzae lipase, Surface Display

References

[1] Sang Yup Lee, Jong Hyun Choi, Zhaohui Xu: TRENDS in Biotechnology Vol.21 (2005), pp.45-52.

[2] GP Smith: Science, Vol. 228 (1985), pp.1315-1317.

[3] Boder Eric, Wittrup K. Dane: Nature biotechnology, Vol.15 (1997), pp.553-557.

[4] Teunissen A W: Yeast, Vol.9 (1993), pp.423-427.

[5] Tanino T, Fukuda H, Kondo A: Biotechnology Progress, Vol.22 (2006), pp.989-993.

[6] Y. d. Wang, X. y. Shen, F. Wang: Journal of Molecular Catalysis B: Enzymatic Vol.67 (2010), pp.45-51.