Purification and on-column activation of a recombinant histidine-tagged pro-transglutaminase after soluble expression in Escherichia coli.
Microbial transglutaminase (MTG) is widely used as a protein crosslinking enzyme. Pro-transglutaminase from Streptomyces mobaraensis was expressed in Escherichia coli as a fusion protein carrying a C-terminal histidine tag (pro-MTG-His(6)) under high-density culture. A new method of on-column activation was designed for production. According to SDS-PAGE, 88.9% of pro-MTG-His(6) was transferred to mature MTG-His(6) with storage stabilization.
Yang
Hui-Lin
HL
College of Bioscience and Bioengineering, South China University of Technology, Guangzhou, China.
Pan
Li
L
Lin
Ying
Y
D000818:Animals; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D004792:Enzyme Precursors / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D006639:Histidine; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000378:metabolism; D012995:Solubility; D013302:Streptomyces / Q000201:enzymology; D011503:Transglutaminases / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D014357:Trypsin / Q000378:metabolism
D004792:Enzyme Precursors; D011994:Recombinant Proteins; D012710:Serum Albumin, Bovine; D006639:Histidine; D011503:Transglutaminases; D014357:Trypsin