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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Characterization of Aldolase from Methanococcus jannaschii by Gas Chromatography

  • Published : 2007.09.30
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Abstract

The products of reactions catalyzed by Methanococcus. jannaschii (Mj) aldolase using various substrates were identified by gas chromatography (GC). Although Mj aldolase is considered a fuculose-1-phosphate aldolase based on homology searching after gene sequencing, it has not been proven to be a fuculose-1-phosphate aldolase based on its reaction products. Mj aldolase was found to catalyze reactions between glycoaldehyde or D, L-glyceraldehyde and DHAP (dihydroxyacetone phosphate). Before performing GC the ketoses produced were converted into peracetylated alditol derivatives by sequential reactions, i.e., dephosphorylation, $NaBH_4$ reduction, and acetylation. By comparing the GC data of final products with those of standard alditol samples, it was found that the enzymatic reactions with glycoaldehyde, D-glyceraldehyde, and D, L-glyceraldehyde produced D-ribulose-1-phosphate, D-psicose-1-phosphate, and a mixture of D-psicose and L-tagatose-1-phosphate, respectively. These results provide direct evidence that Mj aldolase is a fuculose-1-phosphate aldolase.

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References

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