Effect of location of the His-tag on the production of soluble and functional Buthus martensii Karsch insect toxin.
The low yield and poor folding efficiency in vivo of soluble and active recombinant cysteine-rich proteins expressed in Escherichia coli are a major challenge for large-scale protein production and purification. Expression vectors containing Buthus martensii Karsch insect toxin (BmK IT) fused to the C terminus of the intein Ssp DnaB were constructed in an attempt to overcome this problem. Following purification and intein self-cleavage, the fusion protein His(6)-intein-IT produced insoluble BmK IT, while intein-IT-His(6) generated soluble and properly folded BmK IT. This result indicated that the positioning of the His(6) tag has a key role in the production of soluble and functional BmK IT.
Xu
Cheng-Gang
CG
Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Institute of Biotechnology, Shanxi University, Wucheng Road 36, Taiyuan, Shanxi 030006, PR China.
Fan
Xiao-Jun
XJ
Fu
Yue-Jun
YJ
Liang
Ai-Hua
AH
D000818:Animals; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D006639:Histidine / Q000737:chemistry*; D006358:Hot Temperature; D047668:Inteins / Q000502:physiology*; D009842:Oligopeptides / Q000737:chemistry*; D010084:Oxidation-Reduction; D017510:Protein Folding; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012604:Scorpion Venoms / Q000096:biosynthesis*
C509305:Bmk IT toxin, Buthus martensii Karsch; C471213:His-His-His-His-His-His; D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; D012604:Scorpion Venoms; D006639:Histidine