Extracellular metalloprotease gene of Streptomyces cacaoi: structure, nucleotide sequence and characterization of the cloned gene product

Poa Chun Chang, Tai Chih Kuo, Akira Tsugita, Yan Hwa Wu Lee

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

The gene (npr) encoding an extracellular neutral metalloprotease (Npr) from Streptomyces cacaoi YM15 was cloned in Streptomyces lividans using pIJ702 as a vector. The nucleotide (nt) sequence of npr was determined. The deduced open reading frame encoded 550 amino acids (aa)(60 kDa) with a putative signal sequence of 34 aa at the N terminus. High-resolution S1 mapping identified the transcriptional start point at about 132-134 nt upstream from the start codon. The nt sequences at both -10 and -35 regions resemble the consensus sequence of typical Escherichia coli promoters and a fragment containing the promoter was functional in an E. coli promoter probe plasmid. In vitro transcription and translation of the cloned npr sequence revealed a 60-kDa protein product, correlated with the sequence data but not with the size (35 kDa) of the extracellular Npr. The N-terminal as sequence in conjunction with the aa composition analyses on the purified mature Npr led to the conclusion that it was processed from the 60-kDa pre-proenzyme form encoded by npr. The Npr protease contained putative zinc ligand-binding regions and two repeated motifs. Asp-Ser-Gly, similar to the active site residues of the aspartic acid and retroviral proteases.

Original languageEnglish
Pages (from-to)87-95
Number of pages9
JournalGene
Volume88
Issue number1
DOIs
Publication statusPublished - Mar 30 1990
Externally publishedYes

Fingerprint

Streptomyces
Metalloproteases
Amino Acids
Streptomyces lividans
Escherichia coli
Genes
Aspartic Acid Proteases
Enzyme Precursors
Initiator Codon
Consensus Sequence
Protein Sorting Signals
Open Reading Frames
Catalytic Domain
Plasmids
Nucleotides
Proteins

Keywords

  • gene expression
  • in vitro transcription and translation
  • pre-proenzyme
  • Recombinant DNA
  • S1 mapping
  • secretion
  • signal sequence
  • zinc-binding region

ASJC Scopus subject areas

  • Genetics

Cite this

Extracellular metalloprotease gene of Streptomyces cacaoi : structure, nucleotide sequence and characterization of the cloned gene product. / Chang, Poa Chun; Kuo, Tai Chih; Tsugita, Akira; Lee, Yan Hwa Wu.

In: Gene, Vol. 88, No. 1, 30.03.1990, p. 87-95.

Research output: Contribution to journalArticle

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