title: Expression, purification, and characterization of arginine kinase from the sea cucumber Stichopus japonicus.
abstract: 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.
authors.affiliation: Department of Biological Science and Biotechnology, Tsinghua University, Beijing 100084, China.
authors.forename: Shu-Yuan
authors.initials: SY
authors.lastname: Guo
authors.forename: Zhi
authors.initials: Z
authors.lastname: Guo
authors.forename: Qin
authors.initials: Q
authors.lastname: Guo
authors.forename: Bao-Yu
authors.initials: BY
authors.lastname: Chen
authors.forename: Xi-Cheng
authors.initials: XC
authors.lastname: Wang
mesh_terms: 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
chemical_list: D001122:Arginine Kinase