A CMP-N-acetylneuraminic Acid Synthetase Purified from a Marine Bacterium, Photobacterium leiognathi JT-SHIZ-145

• Published in: Bioscience, biotechnology, and biochemistry biotechnology, and biochemistry, 2011-01, Vol.75 (1), p.47-53
• Main Authors: KAJIWARA, Hitomi, MINE, Toshiki, MIYAZAKI, Tatsuo, YAMAMOTO, Takeshi
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; cytidine 5′-monophospho-N-acetylneuraminic acid (CMP-Neu5Ac) synthetase; Escherichia coli; Fundamental and applied biological sciences. Psychology; Kinetics; N-acetylneuraminic acid; N-Acylneuraminate Cytidylyltransferase - biosynthesis; N-Acylneuraminate Cytidylyltransferase - isolation & purification; N-Acylneuraminate Cytidylyltransferase - metabolism; N-glycolylneuraminic acid; Oligosaccharides - analysis; Oligosaccharides - biosynthesis; Oligosaccharides - isolation & purification; Photobacterium - classification; Photobacterium - enzymology; Photobacterium - metabolism; Photobacterium leiognathi; Sialyltransferases - metabolism; Substrate Specificity
• ISSN: 0916-8451; 1347-6947; 1347-6947
• DOI: 10.1271/bbb.100506

A cytidine 5′-monophospho-N-acetylneuraminic acid (CMP-Neu5Ac) synthetase was found in a crude extract prepared from Photobacterium leiognathi JT-SHIZ-145, a marine bacterium that also produces a β-galactoside α2,6-sialyltransferase. The CMP-Neu5Ac synthetase was purified from the crude extract of the cells by a combination of anion-exchange and gel filtration column chromatography. The purified enzyme migrated as a single band (60 kDa) on sodium dodecylsulfate-polyacrylamide gel electrophoresis. The activity of the enzyme was maximal at 35 °C at pH 9.0, and the synthetase required Mg 2+ for activity. Although these properties are similar to those of other CMP-Neu5Ac synthetases isolated from bacteria, this synthetase produced not only CMP-Neu5Ac from cytidine triphosphate and Neu5Ac, but also CMP-N-glycolylneuraminic acid from cytidine triphosphate and N-glycolylneuraminic acid, unlike CMP-Neu5Ac synthetase purified from Escherichia coli.