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A mini-scale method for assay of D-amino acid oxidase activity inTrigonopsis variabilisagainst cephalosporin C

Vijay Chintaman Sonawane

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Abstract

A mini-scale method for the estimation of D-amino acid oxidase activity against cephalosporin C was developed. The method allows implementation of the oxygenation and thorough mixing in a microfuge tube scale reactor. Dilution of reaction products in a buffer containing 0.03% hydrogen peroxide converted the intermediate product of the enzymatic reaction, α-keto adipyl-7-aminocephalosopranic acid (KA-7-ACA) to glutaryl-7-aminocephalosporanic acid (GL-7-ACA). In combination with high performance liquid chromatography, D-amino acid oxidase activity in the whole cells of Trigonopsis variabilis could be measured based on cephalosporin consumed in the reaction or the GL-7-ACA formed. This method could also be used for assay of D-amino acid oxidase activity in crude and soluble preparations.

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What this paper is about

A mini-scale method for the estimation of D-amino acid oxidase activity against cephalosporin C was developed. The method allows implementation of the oxygenation and thorough mixing in a microfuge tube scale reactor. Dilution of reaction products in a buffer containing 0.03% hydrogen peroxide converted the intermediate product of the enzymatic reaction, α-keto adipyl-7-aminocephalosopranic acid (KA-7-ACA) to glutaryl-7-aminocephalosporanic acid (GL-7-ACA). In combination with high performance liquid chromatography, D-amino acid oxidase activity in the whole cells of Trigonopsis variabilis could be measured based on cephalosporin consumed in the reaction or the GL-7-ACA formed. This method could also be used for assay of D-amino acid oxidase activity in crude and soluble preparations.

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Available abstract

A mini-scale method for the estimation of D-amino acid oxidase activity against cephalosporin C was developed. The method allows implementation of the oxygenation and thorough mixing in a microfuge tube scale reactor. Dilution of reaction products in a buffer containing 0.03% hydrogen peroxide converted the intermediate product of the enzymatic reaction, α-keto adipyl-7-aminocephalosopranic acid (KA-7-ACA) to glutaryl-7-aminocephalosporanic acid (GL-7-ACA). In combination with high performance liquid chromatography, D-amino acid oxidase activity in the whole cells of Trigonopsis variabilis could be measured based on cephalosporin consumed in the reaction or the GL-7-ACA formed. This method could also be used for assay of D-amino acid oxidase activity in crude and soluble preparations.

Key concepts: D-amino acid oxidase, Chemistry, Cephalosporin C, Oxidase test, Hydrogen peroxide, Chromatography, Cephalosporin, Amino acid

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A mini-scale method for assay of D-amino acid oxidase activity inTrigonopsis variabilisagainst cephalosporin C — Research Paper | ScholarLens