Patent ID: 6620616
Filing Date: 2003-09-16
Classification: C12N,C12Y

Abstract:
An isolated polynucleotide encoding a variant of a corresponding wild-type nitric oxide synthase,wherein the corresponding wild-type nitric oxide synthase comprises an oxygenase domain having a cysteine residue that binds heme, an alpha three helix, and a tryptohan residue that is located on the alpha 3 helix, six residues upstream from the cysteine residue that binds heme, a reductase domain having an NADPH binding site, and a calmodulin binding region; wherein said variant is a substitution mutant of the corresponding wild-type nitric oxide synthase in which the tryptophan residue that is located on the alpha 3 helix, six residues upstream from the cysteine that binds heme in the corresponding wild-type sequence is substituted with a tyrosine or phenylalanine residue and in which 0 to 10% of the other amino acids in the corresponding wild-type nitric oxide synthase are substituted with conservative amino acids, and wherein said variant has an in vitro enzymatic activity that is greater than the in vitro enzymatic activity of the corresponding wild-type nitric oxide synthase.