Patent ID: 6303360
Filing Date: 2001-10-16
Classification: A23J,C12N,C12P

Abstract:
An isolated nucleic acid sequence encoding a polypeptide having aminopeptidase activity obtained from a Sphingomonas strain, selected from the group consisting of:(a) a nucleic acid sequence encoding a polypeptide having an amino acid sequence which has at least 95% identity with amino acids 33 to 674 of SEQ ID NO:2;(b) a nucleic acid sequence which hybridizes under medium stringency conditions with (i) nucleotides 670 to 2592 of SEQ ID NO:1, (ii) its complementary strand, or (iii) a fragment thereof of at least 100 contiguous nucleotides of nucleotides 670 to 2592 of SEQ ID NO:1, wherein medium stringency conditions are defined as prehybndization and hybridization at 42.degree. C. in 5.times.SSPE, 0.3% SDS, 200 .mu.g/ml sheared and denatured salmon sperm DNA and 35% formamide and washing with 2.times.SSC, 0.2% SDS at 55.degree. C.(c) a nucleic acid sequence encoding an enzymatically active fragment of (a) or (b); and(d) a nucleic adid sequence encoding a polypeptide which (i) has aminopeptidase actvity in the pH range between pH 5.0-8.5 measured at 37.degree. C., (ii) has an isoelectric point in the range of 7.4-8.5, (iii) has aminopeptidase activity in the temperature range of 20-55.degree. C., measured at pH 7.5 using Gly-para-nitroanilide in Tris-HCl buffer, (iv) hydrolyzes Ala-para-nitroanilide, Gly-para-nitroanilide, Leu-para-nitroanilide, Glu-para-nitroanilide, Asp-para-nitroanilide, Lys-para-nitroanilide, Ile-para-nitroanilde and Val-para-nitroanilide; (v) does not hydrolyze Phe-para-nitroanilide or Pro-para-nitroanilide; (vi) is not inhibited by phenylmethanesulfonyl fluoride; and (vii) has a molecular mass of 67.+-.5 kDa by SDS-PAGE.