Expression, purification, and characterization of arginine kinase from the sea cucumber Stichopus japonicus.
The arginine kinase gene of sea cucumber Stichopus japonicus was cloned and inserted into the prokaryotic expression plasmid pET-21b. The protein was expressed in a soluble and functional form in Escherichia coli and purified by Blue Sepharose CL-6B, DEAE-32, and Sephadex G-100 chromotography with a final yield of 83 mgL(-1) of LB medium. The specific activity, electrophoretic mobility, and isoelectric focusing were all identical with those of arginine kinase that was purified from sea cucumber muscle. The fluorescence emission spectrum of arginine kinase had a maximum fluorescence at a wavelength of 330 nm upon excitation at 295 nm. These results are the first report of this purified protein.
Guo
Shu-Yuan
SY
Department of Biological Science and Biotechnology, Tsinghua University, Beijing 100084, China.
Guo
Zhi
Z
Guo
Qin
Q
Chen
Bao-Yu
BY
Wang
Xi-Cheng
XC
D000818:Animals; D001122:Arginine Kinase / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D002848:Chromatography, DEAE-Cellulose; D002942:Circular Dichroism; D004926:Escherichia coli / Q000378:metabolism; D015870:Gene Expression; D007525:Isoelectric Focusing; D010957:Plasmids / Q000235:genetics; D012616:Sea Cucumbers / Q000201:enzymology*; D013050:Spectrometry, Fluorescence
D001122:Arginine Kinase