Development of a satisfactory and general continuous assay for aminotransferases by coupling with (R)-2-hydroxyglutarate dehydrogenase

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Why is it important?

In the context of studying and varying (by mutagenesis) aminotransferase specificity it is essential to have a good continuous assay. Using glutamate dehydrogenase to monitor glutamate production iis problematical and so instead we use a bacterial dehydrogenase that reduces oxoglutarate produced in the amination of keto substrates. This works very well and has allowed us to carry out detailed kinetic studies of rates of production of various amino acids by the branched-chain aminotransferase.