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Information on EC 3.4.24.28 - bacillolysin and Organism(s) Geobacillus stearothermophilus and UniProt Accession P43133

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     3 Hydrolases
         3.4 Acting on peptide bonds (peptidases)
             3.4.24 Metalloendopeptidases
                3.4.24.28 bacillolysin
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This record set is specific for:
Geobacillus stearothermophilus
UNIPROT: P43133 not found.
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The taxonomic range for the selected organisms is: Geobacillus stearothermophilus
The expected taxonomic range for this enzyme is: Bacteria, Archaea, Eukaryota
Reaction Schemes
similar, but not identical, to that of thermolysin
Synonyms
neutral protease, neutrase, protease n, bacillolysin, thermostable neutral protease, bacillus subtilis neutral protease, bae16, bacillus subtilis neutral proteinase, mprbi, bl-ma, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Anilozyme P 10
-
-
-
-
Bacillus metalloendopeptidase
-
-
-
-
Bacillus metalloproteinase
-
-
-
-
Bacillus neutral proteinase
-
-
-
-
Bacillus subtilis neutral proteinase
-
-
-
-
MCP 76
-
-
-
-
Megateriopeptidase
-
-
-
-
Neutral protease
non-specific neutral protease
-
-
Thermostable neutral protease
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY hide
76774-43-1
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2-aminobenzoyl-AGLA-4-nitrobenzylamide + H2O
?
show the reaction diagram
-
-
-
-
?
2-aminobenzoyl-Ala-Gly-Leu-Ala-4-nitrobenzylamide + H2O
?
show the reaction diagram
-
-
-
-
?
3-(2-furylacryloyl)-glycyl-L-leucinamide + H2O
?
show the reaction diagram
-
-
-
-
?
N-(3-[2-furyl]acryloyl)-Gly-Leu amide + H2O
?
show the reaction diagram
-
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
-
5 mM Ca2+ stabilizes the enzyme
Zn2+
-
Zn2+ ions, although essential cofactors and exhibiting extremely high affinity, are without effect on the stability
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
guanidine hydrochloride
-
competitive inhibitor
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1000
guanidine hydrochloride
-
pH 7.5, 25°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
242
-
wild type enzyme at temperature optimum 75°C
273
-
pseudo wild type at temperature optimum 75°C
375
-
double mutant G8C/N60C at temperature optimum 80°C
608
-
boilysin at temperature optimum 95°C
78.5
-
Boilysin at 37°C with casein as substrate
81.5
-
pseudo wild type at 37°C with casein as substrate
81.9
-
wild type enzyme at 37°C with casein as substrate
82.4
-
double mutant G8C/N60C at 37°C with casein as substrate
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5
sharp decrease in activity below
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
60
-
activity toward N-(3-[2-furyl]acryloyl)-Gly-Leu amide is 4 times higher than at 25°C
80
-
double mutant G8C/N60C
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
THER_GEOSE
551
1
60617
Swiss-Prot
-
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
35000
x * 35000, SDS-PAGE
34500
-
Bacillus stearothermophilus NCIB 8924
35000
-
Bacillus stearothermophilus NRRL B-3880
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
G8C/N60C
W55F
-
higher Ca2+ concentration is needed than for wild-type for occupation of Ca2+-binding site III at amino acids 55-59
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
65
1 h, 80% residual activity
65
-
Bacillus stearothermophilus NCIB 8924 enzyme stable at
70
-
Bacillus stearothermophilus B-3880 enzyme stable at
92.5
-
G8C/N60C double mutation extremely thermostable, half-life 35.9 min
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
native wild type and G8C/N60C enzymes are stable toward autoproteolysis and keep their structural integrity for several weeks
-
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2-propanol
-
48 h, 37°C, 50% residual activity with 44.5% (v/v) and 65.5% (v/v) for pseudo wild-type and mutant G8C/N60C respectively
Acetone
-
48 h, 37°C, 50% residual activity with 30.0% (v/v) and 50.1% (v/v) for pseudo wild-type and mutant G8C/N60C respectively
acetonitrile
-
48 h, 37°C, 50% residual activity with 30.6% (v/v) and 51.6% (v/v) for pseudo wild-type and mutant G8C/N60C respectively
dimethylformamide
-
48 h, 37°C, 50% residual activity with 29.8% (v/v) and 45.8% (v/v) for pseudo wild-type and mutant G8C/N60C respectively
dioxane
-
48 h, 37°C, 50% residual activity with 32.1% (v/v) and 47.9% (v/v) for pseudo wild-type and mutant G8C/N60C respectively
DMSO
-
48 h, 37°C, 50% residual activity with 32.9% (v/v) and 48.7% (v/v) for pseudo wild-type and mutant G8C/N60C respectively
Methanol
-
48 h, 37°C, 50% residual activity with 56.6% (v/v) and 86.4% (v/v) for pseudo wild-type and mutant G8C/N60C respectively
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
after expression in Escherichia coli with N-terminal His-tag
-
wild-type and mutant enzymes
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression in Bacillus subtilis
nucleotide sequence and promoter region
-
RENATURED/Commentary
ORGANISM
UNIPROT
LITERATURE
protease activity can be significantly regained if the completely unfolded enzyme is transferred from 8 M guanidine-HCl into native-like conditions (0.4 M guanidine-HCl) by dilution with buffer
-
renaturation of enzyme from inclusion bodies after solubilization using guanidine hydrochloride
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis
-
production of enzyme as an active mature enzyme in Escherichia coli in the absence of its prosequence. Expression of enzyme with N-terminal His-tag, which accumulates as inclusion bodies and renaturation after solubilization using guanidine hydrochloride
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Takagi, M.; Imanaka, T.; Aiba, S.
Nucleotide sequence and promoter region for the neutral protease gene from Bacillus stearothermophilus
J. Bacteriol.
163
824-831
1985
Geobacillus stearothermophilus
Manually annotated by BRENDA team
Sidler, W.; Zuber, H.
Isolation procedures for thermostable neutral proteinases produced by Bacillus stearothermophilus
Eur. J. Appl. Microbiol. Biotechnol.
10
197-209
1980
Geobacillus stearothermophilus
-
Manually annotated by BRENDA team
Sidler, W.; Zuber, H.
Neutral proteases with different thermostabilities from a facultative strain of Bacillus stearothermophilus grown at 40.deg. and at 50.deg
FEBS Lett.
25
292-294
1972
Geobacillus stearothermophilus
Manually annotated by BRENDA team
Duerrschmidt, P.; Mansfeld, J.; Ulbrich-Hofmann, R.
Differentiation between conformational and autoproteolytic stability of the neutral protease from Bacillus stearothermophilus containing an engineered disulfide bond
Eur. J. Biochem.
268
3612-3618
2001
Geobacillus stearothermophilus
Manually annotated by BRENDA team
Duerrschmidt, P.; Mansfeld, J.; Ulbrich-Hofmann, R.
An engineered disulfide bridge mimics the effect of calcium to protect neutral protease against local unfolding
FEBS J.
272
1523-1534
2005
Geobacillus stearothermophilus
Manually annotated by BRENDA team
Mansfeld, J.; Petermann, E.; Duerrschmidt, P.; Ulbrich-Hofmann, R.
The propeptide is not required to produce catalytically active neutral protease from Bacillus stearothermophilus
Protein Expr. Purif.
39
219-228
2005
Geobacillus stearothermophilus
Manually annotated by BRENDA team
Mansfeld, J.; Ulbrich-Hofmann, R.
The stability of engineered thermostable neutral proteases from Bacillus stearothermophilus in organic solvents and detergents
Biotechnol. Bioeng.
97
672-679
2007
Geobacillus stearothermophilus
Manually annotated by BRENDA team
Zhang, M.; Zhao, C.; Du, L.; Lu, F.; Gao, C.
Expression, purification, and characterization of a thermophilic neutral protease from Bacillus stearothermophilus in Bacillus subtilis
Sci. China C Life Sci.
51
52-59
2008
Geobacillus stearothermophilus (P43133), Geobacillus stearothermophilus
Manually annotated by BRENDA team
Duerrschmidt, P.; Mansfeld, J.; Ulbrich-Hofmann, R.
Refolding of the non-specific neutral protease from Bacillus stearothermophilus proceeds via an autoproteolytically sensitive intermediate
Biophys. Chem.
147
66-73
2010
Geobacillus stearothermophilus
Manually annotated by BRENDA team