The enzymic cleavage of N-acetylneuraminic acid

Brug, J.; Rj, Esser; Gb, Paerels

doi:10.1016/0006-3002(59)90521-9

Cited by 13 publications

(3 citation statements)

References 7 publications

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“…3.3] cleaves N-acylneuraminic acid in a reversible reaction to yield pyruvate and /V-acyl-Dmannosamine. This enzyme was originally found in some neuraminidase-producing bacteria such as Vibrio cholerae (1,2) and Clostridium perfringens (3)(4)(5). The enzyme was also found to be widely distributed in animal tissues such as rat liver and brain (5), pig kidney (6), and bovine kidney (7).…”

mentioning

confidence: 99%

Purification and Properties of N-Acetylneuraminate Lyase from Escherichia coli1

Uchida¹,

Tsukada²,

Sugimori³

1984

The Journal of Biochemistry

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N-Acetylneuraminate lyase [N-acetylneuraminic acid aldolase EC 4.1.3.3] from Escherichia coli was purified by protamine sulfate treatment, fractionation with ammonium sulfate, column chromatography on DEAE-Sephacel, gel filtration on Ultrogel AcA 44, and preparative polyacrylamide gel electrophoresis. The purified enzyme preparation was homogeneous on analytical polyacrylamide gel electrophoresis, and was free from contaminating enzymes including NADH oxidase and NADH dehydrogenase. The enzyme catalyzed the cleavage of N-acetylneuraminic acid to N-acetylmannosamine and pyruvate in a reversible reaction. Both cleavage and synthesis of N-acetylneuraminic acid had the same pH optimum around 7.7. The enzyme was stable between pH 6.0 to 9.0, and was thermostable up to 60 degrees C. The thermal stability increased up to 75 degrees C in the presence of pyruvate. No metal ion was required for the enzyme activity, but heavy metal ions such as Ag+ and Hg2+ were potent inhibitors. Oxidizing agents such as N-bromosuccinimide, iodine, and hydrogen peroxide, and SH-inhibitors such as p-chloromercuribenzoic acid and mercuric chloride were also potent inhibitors. The Km values for N-acetylneuraminic acid and N-glycolylneuraminic acid were 3.6 mM and 4.3 mM, respectively. Pyruvate inhibited the cleavage reaction competitively; Ki was calculated to be 1.0 mM. In the condensation reaction, N-acetylglucosamine, N-acetylgalactosamine, glucosamine, and galactosamine could not replace N-acetylmannosamine as substrate, and phosphoenolpyruvate, lactate, beta-hydroxypyruvate, and other pyruvate derivatives could not replace pyruvate as substrate. The molecular weight of the native enzyme was estimated to be 98,000 by gel filtration methods. After denaturation in sodium dodecyl sulfate or in 6 M guanidine-HCl, the molecular weight was reduced to 33,000, indicating the existence of 3 identical subunits. The enzyme could be used for the enzymatic determination of sialic acid; reaction conditions were devised for determining the bound form of sialic acid by coupling neuraminidase from Arthrobacter ureafaciens, lactate dehydrogenase, and NADH.

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confidence: 99%

Purification and Properties of N-Acetylneuraminate Lyase from Escherichia coli1

Uchida¹,

Tsukada²,

Sugimori³

1984

The Journal of Biochemistry

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“…237 In a repeated study, incubating N-acetylneuraminic acid with a V. cholerae filtrate (pH 6.1, 70 h, 37°C), D-ManNAc, but not D-GlcNAc, was found. 238 The earlier observation of splitting into D-GlcNAc and pyruvate 227 was also attributable to a paper chromatography protocol that could not separate D-GlcNAc and D-ManNAc. 238 In another report, the system D-ManNAc/ phosphoenolpyruvate/ATP/rat liver extract (not D-GlcNAc or D-GalNAc; not pyruvic acid) was described for the synthesis of N-acetylneuraminic acid.…”

Section: The Battle For the Stereochemistry 41 The C5 C6 C7 And Cmentioning

confidence: 99%

“…238 The earlier observation of splitting into D-GlcNAc and pyruvate 227 was also attributable to a paper chromatography protocol that could not separate D-GlcNAc and D-ManNAc. 238 In another report, the system D-ManNAc/ phosphoenolpyruvate/ATP/rat liver extract (not D-GlcNAc or D-GalNAc; not pyruvic acid) was described for the synthesis of N-acetylneuraminic acid. 239 Finally, several years later a protocol was described to determine the absolute configuration of N-acetylneuraminic acid by converting it into D-arabinose by chemical degradation.…”

Section: The Battle For the Stereochemistry 41 The C5 C6 C7 And Cmentioning

confidence: 99%

Exploration of the Sialic Acid World

Schauer

Kamerling

2018

Advances in Carbohydrate Chemistry and Biochemistry

252

225

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The Sialic Acid World Table 1 Survey of Reported Structures of Naturally Occurring Members of the Sialic Acid Family-cont'd Name Abbreviation References5-N-Acetyl-8-O-acetyl-9-Olactyl-neuraminic acid Neu5,8Ac 2 9Lt 45 5-N-Acetyl-8-O-methylneuraminic acid Neu5Ac8Me 6,19,25,33,46 5-N-Acetyl-4-O-acetyl-8-Omethyl-neuraminic acid Neu4,5Ac 2 8Me 19 5-N-Acetyl-9-O-acetyl-8-Omethyl-neuraminic acid Neu5,9Ac 2 8Me 6,19,33,46 5-N-Acetyl-9-O-methylneuraminic acid Neu5Ac9Me 47 5-N-Acetyl-4-O-sulfoneuraminic acid Neu5Ac4S 48 5-N-Acetyl-8-O-sulfoneuraminic acid Neu5Ac8S 19,32,49,50 5-N-Acetyl-4-O-acetyl-8-Osulfo-neuraminic acid Neu4,5Ac 2 8S 19 5-N-Acetyl-9-O-phosphoneuraminic acid f,g Neu5Ac9P 51 5-N-Acetyl-2-deoxy-2,3didehydro-neuraminic acid g,h Neu2en5Ac 6,30,33,43,52-54 5-N-Acetyl-9-O-acetyl-2deoxy-2,3-didehydroneuraminic acid g Neu2en5,9Ac 2 55,56 5-N-Acetyl-2-deoxy-2,3didehydro-9-O-lactylneuraminic acid g Neu2en5Ac9Lt 55,565-N-Acetyl-2,7-anhydroneuraminic acid g,i

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Bibliographie Néerlandaise

1959

Recl. Trav. Chim. Pays‐Bas

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The enzymic cleavage of N-acetylneuraminic acid

Cited by 13 publications

References 7 publications

Purification and Properties of N-Acetylneuraminate Lyase from Escherichia coli1

Purification and Properties of N-Acetylneuraminate Lyase from Escherichia coli1

Exploration of the Sialic Acid World

Bibliographie Néerlandaise

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