Patent ID: 6352847
Filing Date: 2002-03-05
Classification: C12N,C12Q

Abstract:
An ammonia elimination reagent in a buffer solution, which comprises isocitrate dehydrogenase, 2-oxoglutaric acid, reduced nicotinamide adenine dinucleotide (NADH), isocitric acid, glutamate dehydrogenase, and magnesium or manganese ions, wherein the isocitrate dehydrogenase is an isocitrate dehydrogenase prepared by a method comprising:(1) preparing an expression vector comprising a modified nucleotide sequence of codon Nos. 1 to 427 in SEQ ID NO:2, wherein the modification is at least one replacement selected from the group consisting of (a) replacement of codon No. 2 (GCC) by a codon selected from the codon group consisting of GCT, GCA, and GCG, and (b) replacement of codon No. 9 (CCC) by a codon selected from the codon group consisting of CCT, CCA, and CCG; (2) transforming a microorganism by using the prepared expression vector; (3) culturing the obtained transformant in a medium; and (4) recovering isocitrate dehydrogenase from the resulting medium.