scholarly journals Cloning and expression in Bacillus subtilis of the npr gene from Bacillus thermoproteolyticus Rokko coding for the thermostable metalloprotease thermolysin

1994 ◽  
Vol 300 (2) ◽  
pp. 599-603 ◽  
Author(s):  
M J O'Donohue ◽  
B P Roques ◽  
A Beaumont
Keyword(s):  
Bacillus Subtilis ◽  
Bacillus Stearothermophilus ◽  
Biochemical Characterization ◽  
Signal Sequence ◽  
Cloning And Expression ◽  
Bacillus Species ◽  
Gene Coding ◽  
Strong Homology ◽  
Different Strains

We report the isolation, cloning and expression, in Bacillus subtilis, of the gene coding for thermolysin, a thermostable metalloprotease which is produced by Bacillus thermoproteolyticus Rokko. The nucleotide sequence has revealed that, like neutral proteases produced by other members of the Bacillus species, thermolysin is probably produced as a preproenzyme carrying a typical N-terminal membrane signal sequence. Further, the thermolysin gene shares a strong homology with two other previously cloned genes from two different strains of Bacillus stearothermophilus. The sequence of the mature secreted protease, inferred from the DNA sequence, is, with two exceptions, identical with the previously published protein sequence of thermolysin [Titani, Hermodson, Ericsson, Walsh and Neurath (1972) Nature (London) 238, 35-37]. The exceptions are Asn37 and Gln119, originally reported to be Asp and Glu respectively. The biochemical characterization of the secreted recombinant protein shows that it is indistinguishable from the wild-type thermolysin.

Crystal structure and biochemical characterization of the transmembrane PAP2 type phosphatidylglycerol phosphate phosphatase from Bacillus subtilis

2017 ◽  
Vol 74 (12) ◽  
pp. 2319-2332 ◽  
Author(s):  
Meriem El Ghachi ◽  
Nicole Howe ◽  
Rodolphe Auger ◽  
Alexandre Lambion ◽  
Annick Guiseppi ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Bacillus Subtilis ◽  
Biochemical Characterization

Structural and biochemical characterization of SpoIIIAF, a component of a sporulation-essential channel in Bacillus subtilis

2018 ◽  
Vol 204 (1) ◽  
pp. 1-8 ◽  
Author(s):  
N. Zeytuni ◽  
K.A. Flanagan ◽  
L.J. Worrall ◽  
S.C. Massoni ◽  
A.H. Camp ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Biochemical Characterization

scholarly journals Biochemical Characterization of the C4-Dicarboxylate Transporter DctA from Bacillus subtilis

2010 ◽  
Vol 192 (11) ◽  
pp. 2900-2907 ◽  
Author(s):  
Maarten Groeneveld ◽  
Ruud G. J. Detert Oude Weme ◽  
Ria H. Duurkens ◽  
Dirk Jan Slotboom
Keyword(s):  
Bacillus Subtilis ◽  
Biochemical Characterization ◽  
Krebs Cycle ◽  
Membrane Vesicles ◽  
Positive Bacterium ◽  
Proton Symport ◽  
Negative Membrane Potential ◽  
Isolated Membrane Vesicles

ABSTRACT Bacterial secondary transporters of the DctA family mediate ion-coupled uptake of C4-dicarboxylates. Here, we have expressed the DctA homologue from Bacillus subtilis in the Gram-positive bacterium Lactococcus lactis. Transport of dicarboxylates in vitro in isolated membrane vesicles was assayed. We determined the substrate specificity, the type of cotransported ions, the electrogenic nature of transport, and the pH and temperature dependence patterns. DctA was found to catalyze proton-coupled symport of the four C4-dicarboxylates from the Krebs cycle (succinate, fumurate, malate, and oxaloacetate) but not of other mono- and dicarboxylates. Because (i) succinate-proton symport was electrogenic (stimulated by an internal negative membrane potential) and (ii) the divalent anionic form of succinate was recognized by DctA, at least three protons must be cotransported with succinate. The results were interpreted in the light of the crystal structure of the homologous aspartate transporter GltPh from Pyrococcus horikoshii.

scholarly journals Characterization of the Bacillus subtilis ywtD Gene, Whose Product Is Involved in γ-Polyglutamic Acid Degradation

2003 ◽  
Vol 185 (7) ◽  
pp. 2379-2382 ◽  
Author(s):  
Takao Suzuki ◽  
Yasutaka Tahara
Keyword(s):  
Escherichia Coli ◽  
Bacillus Subtilis ◽  
Glutamic Acid ◽  
Recombinant Plasmid ◽  
Signal Sequence ◽  
High Molecular Mass ◽  
Enzymatic Analysis ◽  
Polyglutamic Acid ◽  
Partial Similarity

ABSTRACT The ywtD gene, which codes for an enzyme that degrades γ-polyglutamic acid (PGA), was cloned from Bacillus subtilis IFO16449. The gene is located immediately downstream of ywsC and ywtABC, a PGA operon involved in PGA biosynthesis, and it showed partial similarity to genes coding for dl-endopeptidase, a peptidoglycan-degrading enzyme. The ywtD gene, from which signal sequence is excised, was inserted into pET15b, and the recombinant plasmid was then transformed into Escherichia coli. Histidine-tagged YwtD was purified from sonicated cells of the transformant. The purified YwtD degraded PGA to yield two hydrolyzed products, a high-molecular-mass product (490 kDa with nearly 100% l-glutamic acid) and an 11-kDa product (with d-glutamic acid and l-glutamic acid in an 80:20 ratio). This finding and results of enzymatic analysis of the two products with carboxypeptidase G suggest that YwtD is a novel enzyme cleaving the γ-glutamyl bond only between d- and l-glutamic acids of PGA, and it may be designated γ-dl-glutamyl hydrolase.

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