Rui Yamaguchi
   Department   Kumamoto Health Science University  Department of Medical Technology, Faculty of Health Science
   Kumamoto Health Science University Graduate School  Division of Health Sciences, Graduate School of Health Sciences
   Position   Associate Professor
Language English
Publication Date 2015/12
Type
Peer Review Peer reviewed
Title Halophilic starch-binding domain as a novel fusion protein partner for efficient recombinant protein expression
Contribution Type
Journal Bull Soc Sea Water Sci.
Journal TypeJapan
Volume, Issue, Page 69,pp.384-385
Total page number 1
Author and coauthor Yamaguchi R, Ishibashi M, Tokunaga H, Arakawa T, Tokunaga M
Details Halophiles produce halophilic enzymes which function well in high salinity conditions. The starch binding domain(SBD)of halophilic α-amylase has been found to show typical halophilic characteristics with high aqueous solubility and folding capacity. Here we developed a fusion protein expression vector, pSBF-S1, using this SBD as a fusion partner, looking for highly soluble protein expression and efficient affinity-purification. The fusion protein expressed could be purified by dual affinity chromatographies, i.e. Ni-affinity resin with N-terminal His-tag and amylose-resin with SBD affinity. The target protein is then generated through thrombin-digestion of fusion protein at the cleavage site between the target protein and SBD.