... purification of the recombinant enzyme In order to construct an expression vector for Cl-idh, two oligonucleotides (sense, 5¢-AAAAACATATGGCAAGCA AATCGACCATCATCTACAC-3¢, and antisense, 5¢-AAA AAGGATCCCGGCTGAAAACCGGGCTGCATTA-3¢) ... dehydrogenase is known to predominantly catalyze the reduction of oxaloacetate to malate, and thereby lowering the possibilities of oxaloacetate accumulation Indeed, when we have analyzed the malate dehydrogenase ... during the decarboxylation of isocitrate, and indeed, the site-specific mutagenesis of K253 to Met led to an inactive protein [23] In addition, the alkylation of the adjacent M258 inactivated the...