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C1999 Sigma

CMP-Sialic Acid Synthetase from Neisseria meningitidis group B

recombinant, expressed in E. coli BL21, ≥10 units/mg protein

Synonym: CTP: N-Acylneuraminate cytidylyltransferase

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Description

Analysis Note

Enzymatic activity assays are performed in Tris-HCl buffer (100 mM, pH 8.5) containing Neu-5-Ac (1 mM) and CTP (1 mM) at 37 °C for 30 min and analyzed using capillary electrophoresis with a UV detector (200 nm).

Unit Definition

One unit will catalyze the formation of 1 μmol CMP-Neu-5-Ac from Neu-5-Ac and CTP per minute at 37 °C at pH 8.0.

Physical form

Supplied as a lyophilized powder containing Tris-HCl and NaCl.

Application

The enzyme has been utilized to synthesize CMP-sialic acid and its derivatives.1

Protocols & Applications

Sialic Acid Synthesis and Signaling Overview

Price and Availability

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Safety & Documentation

Safety Information

WGK Germany 
3

Protocols & Articles

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Peer-Reviewed Papers

References

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1. Chemoenzymatic synthesis of CMPsialic acid derivatives by a one-pot two-enzyme system: comparison of substrate flexibility of three microbial CMP-sialic acid synthetases. Yu H, et al. Bioorg. Med. Chem. 12, 6427-6435, (2004)

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Purification and characterization of the recombinant CMP-sialic acid synthetase from Neisseria meningitides. Gilbert, M., et al Biotechnol. Lett. 19, 417-420, (1997)

Complete sequencing and characterization of 21,243 full-length human cDNAs. Ota, T. Nat. Genet. 36, 40-5, (2004)

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Glycomimicry: display of the GM3 sugar epitope on Escherichia coli and Salmonella enterica sv Typhimurium. Ilg K, Yavuz E, Maffioli C, et al. Glycobiology 20(10), 1289-97, (2010)

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Genetic engineering of Escherichia coli for the economical production of sialylated oligosaccharides. Fierfort N and Samain E J. Biotechnol. 134(3-4), 261-5, (2008)

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Initial characterization of the human central proteome. Burkard TR, Planyavsky M, Kaupe I, et al. BMC Syst. Biol. 5, 17, (2011)

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Haploid genetic screens in human cells identify host factors used by pathogens. Carette JE, Guimaraes CP, Varadarajan M, et al. Science 326(5957), 1231-5, (2009)

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Site-specific immobilization of CMP-sialic acid synthetase on magnetic nanoparticles and its use in the synthesis of CMP-sialic acid. Yu CC, Lin PC, and Lin CC Chem. Commun. (Camb.) (11), 1308-10, (2008)

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Addition of neuA, the gene encoding N-acylneuraminate cytidylyl transferase, increases the discriminatory ability of the consensus sequence-based scheme for typing Legionella pneumophila serogroup 1 strains. Ratzow S, Gaia V, Helbig JH, et al. J. Clin. Microbiol. 45(6), 1965-8, (2007)

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Sialome analysis of the cephalochordate Branchiostoma belcheri, a key organism for vertebrate evolution. Guérardel Y, Chang LY, Fujita A, et al. Glycobiology 22(4), 479-91, (2012)

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Construction of a functional CMP-sialic acid biosynthesis pathway in Arabidopsis. Castilho A, Pabst M, Leonard R, et al. Plant Physiol. 147(1), 331-9, (2008)

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Deficits in sialylation impair podocyte maturation. Weinhold B, Sellmeier M, Schaper W, et al. J. Am. Soc. Nephrol. 23(8), 1319-28, (2012)

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Bacterial CMP-sialic acid synthetases: production, properties, and applications. Mizanur RM and Pohl NL Appl. Microbiol. Biotechnol. 80(5), 757-65, (2008)

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Stable coexpression of two human sialylation enzymes in plant suspension-cultured tobacco cells. Kajiura H, Misaki R, Fujiyama K, et al. J. Biosci. Bioeng. 111(4), 471-7, (2011)

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The full-ORF clone resource of the German cDNA Consortium. Bechtel S, Rosenfelder H, Duda A, et al. BMC Genomics 8, 399, (2007)

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The rainbow trout CMP-sialic acid synthetase utilises a nuclear localization signal different from that identified in the mouse enzyme. Tiralongo J, Fujita A, Sato C, et al. Glycobiology 17(9), 945-54, (2007)

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Pasteurella multocida CMP-sialic acid synthetase and mutants of Neisseria meningitidis CMP-sialic acid synthetase with improved substrate promiscuity. Li Y, Yu H, Cao H, et al. Appl. Microbiol. Biotechnol. 93(6), 2411-23, (2012)

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Chemoenzymatic synthesis of CMP-N-acetyl-7-fluoro-7-deoxy-neuraminic acid. Hartlieb S, Günzel A, Gerardy-Schahn R, et al. Carbohydr. Res. 343(12), 2075-82, (2008)

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Chemo-enzymatic synthesis of C-9 acetylated sialosides. Rauvolfova J, Venot A, and Boons GJ Carbohydr. Res. 343(10-11), 1605-11, (2008)

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Chemoenzymatic synthesis of mono- and di-fluorinated Thomsen-Friedenreich (T) antigens and their sialylated derivatives. Yan J, Chen X, Wang F, et al. Org. Biomol. Chem. 11(5), 842-8, (2013)

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Cloning and characterization of a heat-stable CMP-N-acylneuraminic acid synthetase from Clostridium thermocellum. Mizanur RM and Pohl NL Appl. Microbiol. Biotechnol. 76(4), 827-34, (2007)

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Identification and biochemical characterization of two functional CMP-sialic acid synthetases in Danio rerio. Schaper W, Bentrop J, Ustinova J, et al. J. Biol. Chem. 287(16), 13239-48, (2012)

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Efficient chemoenzymatic synthesis of sialyl Tn-antigens and derivatives. Ding L, Yu H, Lau K, et al. Chem. Commun. (Camb.) 47(30), 8691-3, (2011)

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Parallel chemoenzymatic synthesis of sialosides containing a C5-diversified sialic acid. Cao H, Muthana S, Li Y, et al. Bioorg. Med. Chem. Lett. 19(20), 5869-71, (2009)

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Identification and characterization of important residues in the catalytic mechanism of CMP-Neu5Ac synthetase from Neisseria meningitidis. Horsfall LE, Nelson A, Berry A. FEBS J. 277, 2779-2790, (2010)

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Identification of the nuclear export signals that regulate the intracellular localization of the mouse CMP-sialic acid synthetase. Fujita A, Sato C, and Kitajima K Biochem. Biophys. Res. Commun. 355(1), 174-80, (2007)

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Cloning and expression of human sialic acid pathway genes to generate CMP-sialic acids in insect cells. Lawrence SM, Huddleston KA, Tomiya N, et al. Glycoconj. J. 18(3), 205-13, (2001)

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Synthesis of sulfone-based nucleotide isosteres: identification of CMP-sialic acid synthetase inhibitors. Wong JH, Sahni U, Li Y, et al. Org. Biomol. Chem. 7(1), 27-9, (2009)

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A C-terminal phosphatase module conserved in vertebrate CMP-sialic acid synthetases provides a tetramerization interface for the physiologically active enzyme. Oschlies M, Dickmanns A, Haselhorst T, et al. J. Mol. Biol. 393(1), 83-97, (2009)

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Direct detection of ligand binding to Sepharose-immobilised protein using saturation transfer double difference (STDD) NMR spectroscopy. Haselhorst T, Münster-Kühnel AK, Oschlies M, et al. Biochem. Biophys. Res. Commun. 359(4), 866-70, (2007)

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A CMP-N-acetylneuraminic acid synthetase purified from a marine bacterium, Photobacterium leiognathi JT-SHIZ-145. Kajiwara H, Mine T, Miyazaki T, et al. Biosci. Biotechnol. Biochem. 75(1), 47-53, (2011)

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The sialic acids. X. Purification and properties of cytidine 5'-monophosphosialic acid synthetase. Kean EL and Roseman S J. Biol. Chem. 241(23), 5643-50, (1966)

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The status, quality, and expansion of the NIH full-length cDNA project: the Mammalian Gene Collection (MGC). Gerhard, DS. Genome Res. 14, 2121-7, (2004)

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