Display of both N- and C-terminal target fusion proteins on the Aspergillus oryzae cell surface using a chitin-binding module

作者：Tabuchi Soichiro; Ito Junji; Adachi Takashi; Ishida Hiroki; Hata Yoji; Okazaki Fumiyoshi; Tanaka Tsutomu; Ogino Chiaki; Kondo Akihiko^*

来源：Applied Microbiology and Biotechnology, 2010, 87(5): 1783-1789.

DOI：10.1007/s00253-010-2664-6

• Cell surface display
• Aspergillus oryzae
• Chitin-binding module
• Triacylglycerol lipase

A novel cell surface display system in Aspergillus oryzae was established by using a chitin-binding module (CBM) from Saccharomyces cerevisiae as an anchor protein. CBM was fused to the N or C terminus of green fluorescent protein (GFP) and the fusion proteins (GFP-CBM and CBM-GFP) were expressed using A. oryzae as a host. Western blotting and fluorescence microscopy analysis showed that both GFP-CBM and CBM-GFP were successfully expressed on the cell surface. In addition, cell surface display of triacylglycerol lipase from A. oryzae (tglA), while retaining its activity, was also successfully demonstrated using CBM as an anchor protein. The activity of tglA was significantly higher when tglA was fused to the C terminus than N terminus of CBM. Together, these results show that CBM used as a first anchor protein enables the fusion of both the N and/or C terminus of a target protein.

• 出版日期2010-8