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PMID: 1886621 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Cloning and expression in yeast of a cDNA clone encoding Aspergillus oryzae neutral protease II, a unique metalloprotease.

Molecular & general genetics : MGG ·Vol. 228 ·No. 1-2 ·1991-08-00 ·Pages 97-103

Tatsumi H, Murakami S, Tsuji RF, Ishida Y, Murakami K, Masaki A, Kawabe H, Arimura H, Nakano E, Motai H

Abstract

The neutral protease II (NpII) from Aspergillus oryzae is a zinc-containing metalloprotease with some unique properties. To elucidate its structure, we isolated a full-length cDNA clone for NpII. Sequence analysis reveals that NpII has a prepro region consisting of 175 amino acids preceding the mature region, which consists of 177 amino acids. As compared with other microbial metalloproteases, NpII is found to be unique in that it shares only a limited homology with them around two zinc ligand His residues and that the positions of the other zinc ligand (Glu) and the active site (His) cannot be established by homology. When a plasmid designed to express the prepro NpII cDNA was introduced into Saccharomyces cerevisiae and the transformant was cultured in YPD medium (2% glucose, 2% polypeptone, 1% yeast extract), it secreted a proNpII. However, in a culture of the same medium containing 0.2 mM ZnCl2, it secreted a mature NpII with a specific activity and N-terminus identical to those of native NpII. This observation suggests that either an autoproteolytic activity or a yeast protease effected the processing.

MeSH Terms
Amino Acid Sequence Aspergillus oryzae/enzymology,genetics Base Sequence Cloning, Molecular Gene Expression Genes, Fungal Metalloendopeptidases/biosynthesis,genetics Molecular Sequence Data Plasmids Saccharomyces cerevisiae/genetics Sequence Homology, Nucleic Acid Transformation, Genetic
Chemicals
Metalloendopeptidases neutral protease II, Aspergillus oryzae
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Tatsumi H
Research and Development Division, Kikkoman Corporation, Chiba, Japan.
Murakami S
Tsuji R F
Ishida Y
Murakami K
Masaki A
Kawabe H
Arimura H
Nakano E
Motai H
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Article Info
Journal
Molecular & general genetics : MGG
Abbr.
Mol Gen Genet
ISSN
0026-8925
DOI
10.1007/BF00282453
Published
1991-08-00
Pages
97-103
Language
English
Region
Germany
NLM ID
0125036
Subset
IM
Databases
GENBANK
S53810, S55712, X56671, X56672, X63982, X63983, X63984, X63985, X63986, X63987
External Links
PubMed source DOI: 10.1007/BF00282453
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