Characterization of the gene encoding the glutamic-acid-specific protease of Streptomyces griseus

1993 ◽  
Vol 71 (9-10) ◽  
pp. 454-461 ◽  
Author(s):  
Sachdev S. Sidhu ◽  
Gabriel B. Kalmar ◽  
Thor J. Borgford
Keyword(s):  
Glutamic Acid ◽  
Serine Protease ◽  
Streptomyces Griseus ◽  
Gene Encoding ◽  
Lysobacter Enzymogenes ◽  
Glutamic Acid Residue ◽  
Specific Protease ◽  
Mature Enzyme ◽  
Additional Segment ◽  
Pro Region

The complete gene sequence (sprE) for the glutamic-acid-specific serine protease (SGPE) of the gram-positive bacterium Streptomyces griseus is reported. The sprE gene encodes a 355 amino acid pre–pro–mature enzyme. The presence of a glutamic acid residue at the junction of the pro and mature segments of the protein suggests that the enzyme is self-processing. SGPE was found to have extensive homology with the S. griseus proteases A and B. However, there is an additional segment to the pro region of SGPE, lacking in proteases A and B, which has significant homology to the pro region of the α-lytic protease of the gram-negative bacterium Lysobacter enzymogenes. Expression of recombinant SGPE in Bacillus subtilis is also reported, and the enzyme is shown to be self-processing.Key words: serine protease, maturation, expression, propeptide.

PCR-based identification ofStaphylococcus epidermidistargetinggseAencoding the glutamic-acid-specific protease

2004 ◽  
Vol 50 (7) ◽  
pp. 493-498 ◽  
Author(s):  
Y Ikeda ◽  
Y Ohara-Nemoto ◽  
S Kimura ◽  
K Ishibashi ◽  
K Kikuchi
Keyword(s):  
Glutamic Acid ◽  
Staphylococcus Epidermidis ◽  
Single Step ◽  
Clinical Isolates ◽  
Rrna Gene ◽  
Pcr Assay ◽  
Coagulase Negative Staphylococci ◽  
Gene Encoding ◽  
Staphylococcus Capitis ◽  
Specific Protease

The frequency of the gseA gene encoding a glutamic acid-specific serine protease, GluSE, of Staphylococcus epidermidis was investigated. DNA hybridization analysis demonstrated that gseA existed exclusively in S. epidermidis but not in other bacteria examined. A single step PCR assay with a set of designed primers yielded amplification of gseA from all 69 clinical isolates of S. epidermidis taken from patients and healthy adults, whereas production of GluSE was observed in 74% (51/69) of the isolates. Furthermore, none of the 46 clinical isolates of other species of coagulase-negative staphylococci and 45 clinical isolates of Staphylococcus aureus showed amplification, except a Staphylococcus capitis strain. However, this strain was positive for a S. epidermidis-specific DNA region and the DNA sequence of the 16S rRNA gene showed 99% identity with that of S. epidermidis. Therefore, these results indicated that the present PCR assay for gseA was ubiquitous and highly specific for detection of S. epidermidis.Key words: Staphylococcus epidermidis, glutamic acid-specific protease, PCR assay, molecular identification.

Isolation and characterization of a gene encoding a chymotrypsin-like serine protease fromStreptomyces lividans66

1996 ◽  
Vol 42 (3) ◽  
pp. 284-288 ◽  
Author(s):  
Craig Binnie ◽  
Linda Liao ◽  
Eva Walczyk ◽  
Lawrence T. Malek
Keyword(s):  
Amino Acids ◽  
Serine Protease ◽  
Streptomyces Griseus ◽  
Nucleotide Sequence Analysis ◽  
Predicted Amino Acid Sequence ◽  
Gene Encoding ◽  
Hybridization Probe ◽  
Putative Gene ◽  
Isolation And Characterization ◽  
High Degree

A gene encoding a Streptomyces lividans homologue of the chymotrypsin-like serine protease (SAM-P20) of Streptomyces albogriseolus was isolated using the Streptomyces griseus prtB gene as a hybridization probe. Nucleotide sequence analysis of a representative clone uncovered the possible presence of a sequence of 900 nucleotides encoding 300 amino acids, including a putative "prepro" region of 115 amino acids. Alignment of the predicted amino acid sequence of this putative gene with other members of the family of Streptomyces extracellular chymotrypsin-like proteases indicated a high degree of homology in all cases, especially with the SAM-P20 protease. This gene product has been identified as the second member of a potentially larger family of SAL (SAM-P20-like) proteases in S. lividans 66. Keywords: Streptomyces, protease, chymotrypsin.

scholarly journals The Identification of a Glutamic Acid Residue as Part of the Active Site of Ribonuclease T1

1967 ◽  
Vol 242 (20) ◽  
pp. 4682-4690 ◽  
Author(s):  
Kenji Takahashi ◽  
William H. Stein ◽  
Stanford Moore
Keyword(s):  
Glutamic Acid ◽  
Active Site ◽  
Ribonuclease T1 ◽  
Glutamic Acid Residue

Mutations in the gene encoding the serine protease inhibitor, Kazal type 1 are associated with chronic pancreatitis

10.1038/76088 ◽  
2000 ◽  
Vol 25 (2) ◽  
pp. 213-216 ◽  
Author(s):  
Heiko Witt ◽  
Werner Luck ◽  
Hans Christian Hennies ◽  
Martin Claßen ◽  
Andreas Kage ◽  
...  
Keyword(s):  
Chronic Pancreatitis ◽  
Protease Inhibitor ◽  
Serine Protease ◽  
Serine Protease Inhibitor ◽  
Gene Encoding

scholarly journals Mirolase, a novel subtilisin-like serine protease from the periodontopathogen Tannerella forsythia

2015 ◽  
Vol 396 (3) ◽  
pp. 261-275 ◽  
Author(s):  
Miroslaw Ksiazek ◽  
Abdulkarim Y. Karim ◽  
Danuta Bryzek ◽  
Jan J. Enghild ◽  
Ida B. Thøgersen ◽  
...  
Keyword(s):  
Serine Protease ◽  
Calcium Ions ◽  
Enzyme Stability ◽  
Amidolytic Activity ◽  
Tannerella Forsythia ◽  
Calcium Dependent ◽  
Genes Encoding ◽  
Mature Enzyme ◽  
Almost All ◽  
Unique Mechanism

Abstract The genome of Tannerella forsythia, an etiological factor of chronic periodontitis, contains several genes encoding putative proteases. Here, we characterized a subtilisin-like serine protease of T. forsythia referred to as mirolase. Recombinant full-length latent promirolase [85 kDa, without its signal peptide (SP)] processed itself through sequential autoproteolytic cleavages into a mature enzyme of 40 kDa. Mirolase latency was driven by the N-terminal prodomain (NTP). In stark contrast to almost all known subtilases, the cleaved NTP remained non-covalently associated with mirolase, inhibiting its proteolytic, but not amidolytic, activity. Full activity was observed only after the NTP was gradually, and fully, degraded. Both activity and processing was absolutely dependent on calcium ions, which were also essential for enzyme stability. As a consequence, both serine protease inhibitors and calcium ions chelators inhibited mirolase activity. Activity assays using an array of chromogenic substrates revealed that mirolase specificity is driven not only by the substrate-binding subsite S1, but also by other subsites. Taken together, mirolase is a calcium-dependent serine protease of the S8 family with the unique mechanism of activation that may contribute to T. forsythia pathogenicity by degradation of fibrinogen, hemoglobin, and the antimicrobial peptide LL-37.

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