Molecular cloning and sequence analysis of the scpZ gene encoding the serine carboxypeptidase of Absidia zychae.
Carboxypeptidase Z is a serine carboxypeptidase secreted by Absidia zychae NRIC 1199. The cDNA and genomic DNA carrying the scpZ gene encoding carboxypeptidase Z were cloned and sequenced. The nucleotide sequences of the cDNA (1.4 kb) and the genomic DNA (3.3 kb) were analyzed and the intervening sequences were located by a comparison of the two. It was found that the scpZ gene was interrupted by 11 short introns, 50-75 nucleotides in length. Genomic Southern analysis showed that there was only one scpZ gene in the genome of A. zychae. The gene encoded a putative pre-pro-enzyme composed of 409 amino-acid residues of the mature carboxypeptidase Z (M(r) 45,421) and an additional N-terminal sequence of 51 amino-acid residues. The amino-acid sequence around the active serine residue of carboxypeptidase Z (-G-E-S-Y-G-G-) differed from the consensus (-G-E-S-Y-A-G-) which is conserved in most of the serine carboxypeptidases so far analyzed.
Lee
B R
BR
Laboratory of Molecular Biology and Microbial Chemistry, Tokyo University of Agriculture and Technology, Japan.
Takeuchi
M
M
Kobayashi
Y
Y
D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D002268:Carboxypeptidases / Q000235:genetics*; D003001:Cloning, Molecular; D017124:Conserved Sequence; D005656:Fungal Proteins / Q000235:genetics; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D009090:Mucorales / Q000235:genetics*; D010940:Plant Proteins / Q000235:genetics; D015183:Restriction Mapping; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D005656:Fungal Proteins; D010940:Plant Proteins; D002268:Carboxypeptidases; C022633:serine carboxypeptidase; C499287:carboxypeptidase Z