Information on EC 3.4.24.75 - lysostaphin

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY
3.4.24.75
-
RECOMMENDED NAME
GeneOntology No.
lysostaphin
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
Hydrolysis of the -Gly-/-Gly- bond in the pentaglycine inter-peptide link joining staphylococcal cell wall peptidoglycans
show the reaction diagram
-
-
-
-
Hydrolysis of the -Gly-/-Gly- bond in the pentaglycine inter-peptide link joining staphylococcal cell wall peptidoglycans
show the reaction diagram
a two-stage reaction controlled by two ionogenic groups of the active site, the active site of lysostaphin contains glutamic acid and lysine
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis of peptide bond
-
-
-
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
ALE-1
-
-
-
-
Glycyl-glycine endopeptidase
-
-
-
-
Glycylglycine endopeptidase
-
-
-
-
Glycylglycine endopeptidase
-
-
Glycylglycine endopeptidase
Lactobacillus fermentum BR11
-
-
-
Glycylglycine endopeptidase
-
-
Glycylglycine endopeptidase
-
-
-
Glycylglycine endopeptidase
-
-
lysostaphin
-
analysis of Bacillus subtilis bacteriophage phi29 gene product 13's lysostaphin activity
lysostaphin
-
-
lysostaphin
-
EC 3.4.99.17
r-lysostaphin
-
-
r-lysostaphin
-
recombinant lysostaphin
Staphylococcus aureus-specific cell wall endopeptidase lysostaphin
-
-
Lysostaphin endopeptidase
-
-
-
-
additional information
-
lysostaphin-type enzymes belong to so-called LAS, i.e. Lysostaphin, D-Ala-D-Ala carboxypeptidase, Sonic hedgehog enzyme, enzymes
CAS REGISTRY NUMBER
COMMENTARY
9011-93-2
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
Lactobacillus fermentum BR11
strain BR11
-
-
Manually annotated by BRENDA team
strain ATCC 14917
-
-
Manually annotated by BRENDA team
subspecies cremoris MG1363
-
-
Manually annotated by BRENDA team
Staphylococcus capitis EPK1
EPK1
-
-
Manually annotated by BRENDA team
biovar Staphylococcus simulans staphylolyticus
-
-
Manually annotated by BRENDA team
biovar staphylolyticus No. 1030
-
-
Manually annotated by BRENDA team
biovar staphylolyticus, ATCC 1362
-
-
Manually annotated by BRENDA team
biovar staphylolyticus, strain GNTs PM No. 1030
-
-
Manually annotated by BRENDA team
biovar staphylolyticus, strain NRRL B-2628
-
-
Manually annotated by BRENDA team
biovariant Staphylococcus simulans staphylolyticus
-
-
Manually annotated by BRENDA team
Staphylococcus simulans biovar staphylolyticus
Uniprot
Manually annotated by BRENDA team
Staphylococcus simulans NRRL B-2628
NRRL B-2628
-
-
Manually annotated by BRENDA team
Staphylococcus simulans TNK1
strain TNK1
-
-
Manually annotated by BRENDA team
strain K-6-WI
-
-
Manually annotated by BRENDA team
Staphylococcus sp. K-6-WI
strain K-6-WI
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
physiological function
-
chimeric phage lysins act synergistically with lysostaphin to kill mastitis-causing Staphylococcus aureus in mouse mammary glands
additional information
-
lysostaphin is effective, in combination with antibiotic ranalexin, against methicillin-resistant Staphylococcus aureus infections in human MRSA252 cell culture for hemolysis assay, the female ICR CD-1 mouse model for systemic infection, and a rabbit model for wound infection using New Zealand White rabbits, overview. Ranalexin alone causes hemolysis and shows signs of toxicity to Vero cells at over 50 mg/L, but lysostaphin alone causes no hemolysis and has no effect on Vero cell viability
additional information
-
lysostaphin clears Staphylococcus aureus biofilms in vitro and also eradicates established Staphylococcus aureus biofilms on implanted jugular vein catheters in mice, overview
additional information
-
lysostaphin is effective, in combination with antibiotic ranalexin, against methicillin-resistant Staphylococcus aureus infections. The combination is significantly more effective than treatment with ranalexin or lysostaphin alone
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
Elastin + H2O
Fragments of elastin
show the reaction diagram
-
-
-
-
-
L-lysine(N-epsilon-pentaglycine-N-(2-aminobenzoyl))-[N-(2,4-dinitrophenyl)ethylenediamine] + H2O
?
show the reaction diagram
-
-
-
-
?
MV11 + H2O
?
show the reaction diagram
-
developing a fluorescence resonance energy transfer substrate based on MV11
-
-
?
N-(2-aminobenzoyl)pentaglycine-[N-(2,4-dinitrophenyl)ethylenediamine] + H2O
?
show the reaction diagram
-
-
-
-
?
N-Acetylhexaglycine + H2O
?
show the reaction diagram
-
-
-
-
-
Peptidoglycan + H2O
Fragments of peptidoglycan
show the reaction diagram
-
-
-
-
?
Peptidoglycan + H2O
Fragments of peptidoglycan
show the reaction diagram
-
-
-
?
Peptidoglycan + H2O
Fragments of peptidoglycan
show the reaction diagram
-
-
-
-
?
Peptidoglycan + H2O
Fragments of peptidoglycan
show the reaction diagram
-
-
-
-
-
Peptidoglycan + H2O
Fragments of peptidoglycan
show the reaction diagram
-
-
-
-
?
Peptidoglycan + H2O
Fragments of peptidoglycan
show the reaction diagram
-
-
-
-
?
Peptidoglycan + H2O
Fragments of peptidoglycan
show the reaction diagram
-
-
-
-
?
Peptidoglycan + H2O
Fragments of peptidoglycan
show the reaction diagram
-
92 amino acids of the C-terminus of lysostaphin are necessary and sufficiant for targeting the substrate
-
-
-
Peptidoglycan + H2O
Fragments of peptidoglycan
show the reaction diagram
-
hydrolyis of pentaglycine interpeptide bridges of Staphylococcal peptidoglycan
-
-
-
Peptidoglycan + H2O
Fragments of peptidoglycan
show the reaction diagram
-
hydrolyis of pentaglycine interpeptide bridges of Staphylococcal peptidoglycan
-
-
-
Peptidoglycan + H2O
Fragments of peptidoglycan
show the reaction diagram
-
activities of three enzymes namely, glycylglycine endopeptidase, endo-beta-N-acetyl glucosamidase and N-acteyl muramyl-L-alanine amidase. Lysostaphin rapidly lyses actively growing and non-dividing cells including staphylococci in biofilms. Glycylglycine endopeptidase lyses staphylococcal cells by hydrolyzing glycylglycine bonds in the polyglycine bridges which form cross links between glycopeptide chains in the cell wall peptidoglycan of Staphylococcus aureus cells. Lytic principle of lysostaphin is a peptidase which liberates N-terminal glycine and alanine from Staphylococcus aureus cell wall. Hexosaminidase is present in lysostaphin preparation is specific for the gluocainyl-muramic acid bond of the bacterial carbohydrate backbone
-
-
-
Peptidoglycan + H2O
Fragments of peptidoglycan
show the reaction diagram
-
cleaves Gly-Gly bonds in peptidoglycans
-
-
?
Peptidoglycan + H2O
Fragments of peptidoglycan
show the reaction diagram
-
the enzyme is highly specific and effective against Staphylococcus aureus cleaving the pentaglycine cross-bridges in the peptidoglycan layer of staphylococcal cell walls
-
-
?
Peptidoglycan + H2O
Fragments of peptidoglycan
show the reaction diagram
Staphylococcus simulans NRRL B-2628
-
-
-
-
-
Peptidoglycan + H2O
Fragments of peptidoglycan
show the reaction diagram
Lactobacillus rhamnosus GG, Lactobacillus fermentum BR11
-
-
-
-
?
Peptidoglycan + H2O
Fragments of peptidoglycan
show the reaction diagram
Staphylococcus capitis EPK1
-
hydrolyis of pentaglycine interpeptide bridges of Staphylococcal peptidoglycan
-
-
-
peptidoglycan + H2O
?
show the reaction diagram
-
the hydrolytic effect on staphylococcal peptidoglycan can be potentiated by simultaneous action of lysostaphin and lysozyme
-
-
-
peptidoglycan + H2O
?
show the reaction diagram
-
The enzymes attack cell walls of staphylococcus species that do not poduce lysostaphin, species which produce lysostaphin are not attacked. Mutants of Staphylococcus simulans lacking lysostaphin are also attacked
-
-
-
soluble elastin + H2O
?
show the reaction diagram
-
-
-
-
?
L-lysine(N-epsilon-pentaglycyl-DABCYL)-EDANS + H2O
?
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
no hydrolysis of beta-casein
-
-
-
additional information
?
-
-
LytM is expressed during early exponential phase and is overexpressed in an autolysis-defective mutant compared with the parent strain. LytM plays a role in cell growth
-
?
additional information
?
-
-
purified cell walls of a passage-selected glycopeptide-intermediate Staphylococcus aureus isolate
-
-
-
additional information
?
-
-
a traP mutant strain is more resistant to lysostaphin than wild-type Staphylococcus aureus, the extracellular C-terminus of TRAP can act as the specific targeting protein of the lysozyme/lysostaphin on the Staphylococcus aureus cell wall and the biological significance of the interaction might be to facilitate lysozyme/lysostaphin-mediated cell lysis, TRAP is necessary for the binding and activity of lysostaphin in vitro, overview
-
-
-
additional information
?
-
-
lysostaphin is a glycylglycine endopeptidase cleaving the pentaglycine cross-bridge structure unique to the staphylococcal cell wall and is considered to be a potential drug for Staphylococcus aureus, in vitro activity of recombinant lysostaphin against Staphylococcus aureus isolates from hospitals in Beijing, China, with MIC values of 30-2000 ng/ml, agar dilution assay, overview
-
-
-
additional information
?
-
-
lysostaphin is active in treatment of neonatal Staphylococcus aureus infection by a clinical methicillin-sensitive Staphylococcus aureus serotype 5 strain, overview
-
-
-
additional information
?
-
-
potent, synergistic inhibition of Staphylococcus aureus strains MSSA476 and MRSA252 upon exposure to a combination of the endopeptidase lysostaphin and the cationic peptide ranalexin, which is most potent, as well as with the antimicrobial peptides magainin 2 and dermaseptin s3(1-16), overview
-
-
-
additional information
?
-
-
Staphylococcus simulans secretes lysostaphin, a bacteriolytic enzyme that specifically binds to the cell wall envelope of Staphylococcus aureus and cleaves the pentaglycine cross bridges of peptidoglycan, thereby killing staphylococci, screening of a collection of Staphylococcus aureus strain Newman transposon mutants for lysostaphin resistance, transposon insertion in SAV2335 is herein named lyrA, i.e. lysostaphin resistance A, activity against diverse Staphylococcus aureus strains with MIC-values of 0.0005-0.002 mg/ml, overview
-
-
-
additional information
?
-
-
the endopeptidase rapidly lyses Staphylococcus aureus through proteolysis of the staphylococcal cell wall which allows its use as a therapeutic agent, the enzyme acts synergistically with oxacillin and with vancomycin, in vivo antimicrobial activity in different mouse tissues, overview
-
-
-
additional information
?
-
-
the enzyme is active against clinical methiciliin-resistant Staphylococcus aureus with a MIC50 value of 0.125 mg/l, it breaks down the cell wall of Staphylococcus aureus, overview
-
-
-
additional information
?
-
-
pentaglycine bridges in peptidoglycan of Staphylococci as a substrate
-
-
-
additional information
?
-
-
the enzyme is a glycylglycine endopeptidase
-
-
-
additional information
?
-
-
following pseudoinfection of sacculi with bacteriophage phi29 and gene protein 13 respectively: release of muropeptides (predominantly p7) into the supernatant, a modest decrease in the cross-linked muropeptide p21, gene product 13 does not cause cleavage of pentaglycine
-
-
?
additional information
?
-
-
lysostaphin exhibits a high degree of antistaphylococcal bacteriolytic activity, being inactive against bacteria of all other genera. The cell-wall degrading activity of lysostaphin is primarily due to a glycylglycine endopeptidase activity. Staphylococcus simulans biovar staphylolyticus peptidoglycan is resistant to the hydrolytic activity of lysostaphin, since the cells produce a resistance factor that causes the incorporation of Ser residues into the third and fifth positions of the cell wall cross bridges
-
-
-
additional information
?
-
-
resistance to lysostaphin is due to a FemABX-like immunity protein that inserts serines in place of some glycines in peptidoglycan cross bridges
-
-
-
additional information
?
-
-
lysostaphin exhibits strong lytic activity against Staphylococcus aureus mastitis isolates
-
-
-
additional information
?
-
Staphylococcus simulans TNK1
-
Staphylococcus simulans secretes lysostaphin, a bacteriolytic enzyme that specifically binds to the cell wall envelope of Staphylococcus aureus and cleaves the pentaglycine cross bridges of peptidoglycan, thereby killing staphylococci, screening of a collection of Staphylococcus aureus strain Newman transposon mutants for lysostaphin resistance, transposon insertion in SAV2335 is herein named lyrA, i.e. lysostaphin resistance A, activity against diverse Staphylococcus aureus strains with MIC-values of 0.0005-0.002 mg/ml, overview
-
-
-
additional information
?
-
Staphylococcus capitis EPK1
-
no hydrolysis of beta-casein
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
Peptidoglycan + H2O
Fragments of peptidoglycan
show the reaction diagram
-
-
-
-
?
Peptidoglycan + H2O
Fragments of peptidoglycan
show the reaction diagram
-
activities of three enzymes namely, glycylglycine endopeptidase, endo-beta-N-acetyl glucosamidase and N-acteyl muramyl-L-alanine amidase. Lysostaphin rapidly lyses actively growing and non-dividing cells including staphylococci in biofilms. Glycylglycine endopeptidase lyses staphylococcal cells by hydrolyzing glycylglycine bonds in the polyglycine bridges which form cross links between glycopeptide chains in the cell wall peptidoglycan of Staphylococcus aureus cells. Lytic principle of lysostaphin is a peptidase which liberates N-terminal glycine and alanine from Staphylococcus aureus cell wall. Hexosaminidase is present in lysostaphin preparation is specific for the gluocainyl-muramic acid bond of the bacterial carbohydrate backbone
-
-
-
Peptidoglycan + H2O
Fragments of peptidoglycan
show the reaction diagram
-
cleaves Gly-Gly bonds in peptidoglycans
-
-
?
Peptidoglycan + H2O
Fragments of peptidoglycan
show the reaction diagram
-
the enzyme is highly specific and effective against Staphylococcus aureus cleaving the pentaglycine cross-bridges in the peptidoglycan layer of staphylococcal cell walls
-
-
?
peptidoglycan + H2O
?
show the reaction diagram
-
the hydrolytic effect on staphylococcal peptidoglycan can be potentiated by simultaneous action of lysostaphin and lysozyme
-
-
-
peptidoglycan + H2O
?
show the reaction diagram
-
The enzymes attack cell walls of staphylococcus species that do not poduce lysostaphin, species which produce lysostaphin are not attacked. Mutants of Staphylococcus simulans lacking lysostaphin are also attacked
-
-
-
additional information
?
-
-
LytM is expressed during early exponential phase and is overexpressed in an autolysis-defective mutant compared with the parent strain. LytM plays a role in cell growth
-
?
additional information
?
-
-
a traP mutant strain is more resistant to lysostaphin than wild-type Staphylococcus aureus, the extracellular C-terminus of TRAP can act as the specific targeting protein of the lysozyme/lysostaphin on the Staphylococcus aureus cell wall and the biological significance of the interaction might be to facilitate lysozyme/lysostaphin-mediated cell lysis, TRAP is necessary for the binding and activity of lysostaphin in vitro, overview
-
-
-
additional information
?
-
-
lysostaphin is a glycylglycine endopeptidase cleaving the pentaglycine cross-bridge structure unique to the staphylococcal cell wall and is considered to be a potential drug for Staphylococcus aureus, in vitro activity of recombinant lysostaphin against Staphylococcus aureus isolates from hospitals in Beijing, China, with MIC values of 30-2000 ng/ml, agar dilution assay, overview
-
-
-
additional information
?
-
-
lysostaphin is active in treatment of neonatal Staphylococcus aureus infection by a clinical methicillin-sensitive Staphylococcus aureus serotype 5 strain, overview
-
-
-
additional information
?
-
-
potent, synergistic inhibition of Staphylococcus aureus strains MSSA476 and MRSA252 upon exposure to a combination of the endopeptidase lysostaphin and the cationic peptide ranalexin, which is most potent, as well as with the antimicrobial peptides magainin 2 and dermaseptin s3(1-16), overview
-
-
-
additional information
?
-
-
Staphylococcus simulans secretes lysostaphin, a bacteriolytic enzyme that specifically binds to the cell wall envelope of Staphylococcus aureus and cleaves the pentaglycine cross bridges of peptidoglycan, thereby killing staphylococci, screening of a collection of Staphylococcus aureus strain Newman transposon mutants for lysostaphin resistance, transposon insertion in SAV2335 is herein named lyrA, i.e. lysostaphin resistance A, activity against diverse Staphylococcus aureus strains with MIC-values of 0.0005-0.002 mg/ml, overview
-
-
-
additional information
?
-
-
the endopeptidase rapidly lyses Staphylococcus aureus through proteolysis of the staphylococcal cell wall which allows its use as a therapeutic agent, the enzyme acts synergistically with oxacillin and with vancomycin, in vivo antimicrobial activity in different mouse tissues, overview
-
-
-
additional information
?
-
-
the enzyme is active against clinical methiciliin-resistant Staphylococcus aureus with a MIC50 value of 0.125 mg/l, it breaks down the cell wall of Staphylococcus aureus, overview
-
-
-
additional information
?
-
-
lysostaphin exhibits a high degree of antistaphylococcal bacteriolytic activity, being inactive against bacteria of all other genera. The cell-wall degrading activity of lysostaphin is primarily due to a glycylglycine endopeptidase activity. Staphylococcus simulans biovar staphylolyticus peptidoglycan is resistant to the hydrolytic activity of lysostaphin, since the cells produce a resistance factor that causes the incorporation of Ser residues into the third and fifth positions of the cell wall cross bridges
-
-
-
additional information
?
-
-
resistance to lysostaphin is due to a FemABX-like immunity protein that inserts serines in place of some glycines in peptidoglycan cross bridges
-
-
-
additional information
?
-
Staphylococcus simulans TNK1
-
Staphylococcus simulans secretes lysostaphin, a bacteriolytic enzyme that specifically binds to the cell wall envelope of Staphylococcus aureus and cleaves the pentaglycine cross bridges of peptidoglycan, thereby killing staphylococci, screening of a collection of Staphylococcus aureus strain Newman transposon mutants for lysostaphin resistance, transposon insertion in SAV2335 is herein named lyrA, i.e. lysostaphin resistance A, activity against diverse Staphylococcus aureus strains with MIC-values of 0.0005-0.002 mg/ml, overview
-
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Na+
-
stimulation
NH4+
-
stimulation
Zinc
-
0.9 mol zinc per mol LytM
Zn2+
-
metalloprotease, 1 mol of Zn2+ per mol of enzyme, atomic absorption spectroscopy
Zn2+
-
the central Zn2+ is tetrahedrally coordinated by two histidines, an aspartate, and a water molecule. The Zn2+ chelating residue occur in the order histidine, aspartate, histidine and contact the metal via the Nepsilon, the Odelta and the Ndelta respectively, H210(Nepsilon), D214(Odelta), H293(Ndelta)
Zn2+
-
general protein folding and Zn2+ binding are maintained in the D195C mutant but reduced in the D195A mutant
Zn2+
-
contains zinc
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
1,10-phenanthroline
-
-
1,10-phenanthroline
-
-
Co2+
-
10 mM, 30% decrease of activity of enzyme produced with pET vector system and 20% decrease of activity of enzyme produced with pBAD vector system
Cu2+
-
2 mM, 70-80% decrease of activity
diammonium sulfate
-
-
diethyldicarbonate
-
-
diethyldicarbonate
-
-
Ni2+
-
2 mM, 60% decrease of activity of enzyme produced with pET vector system and 20% decrease of activity of enzyme produced with pBAD vector system
Zn2+
-
2 mM, 70-80% decrease of activity
additional information
-
not inhibitory is trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane, i.e. E-64
-
additional information
-
binding of elastin to lysostaphin is not affected by exogenously added zinc, suggesting that zinc interferes directly with the catalytic activity of the enzyme
-
additional information
-
no inhibitory effect of EDTA and 2 mM Co2+
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
C-terminus of TRAP
-
i.e. target of RNAIII activating protein, a membrane-associated protein in Staphylococcus sp., can enhance the activity of lysozyme and lysostaphin, interaction analysis, overview
-
additional information
-
the enzyme acts synergistically with oxacillin and with vancomycin in lysis of the cell wall of pathogenic Staphylococcus aureus
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.07
-
L-lysine(N-e-pentaglycine-N-(2-aminobenzoyl))-[N-(2,4-dinitrophenyl)ethylenediamine]
-
value from Grafit and Hanes plot analyses
0.07
-
L-lysine(N-e-pentaglycyl-DABCYL)-EDANS
-
value from Hanes plot analysis
0.2
-
N-(2-Aminobenzoyl)pentaglycine-[N-(2,4-dinitrophenyl)ethylenediamine]
-
value from Grafit plot analysis
0.3
-
N-(2-Aminobenzoyl)pentaglycine-[N-(2,4-dinitrophenyl)ethylenediamine]
-
value from Hanes plot analysis
0.08
-
L-lysine(N-e-pentaglycyl-DABCYL)-EDANS
-
value from Grafit plot analysis
additional information
-
additional information
-
kinetics and thermodynamics
-
additional information
-
additional information
-
kinetics of lysostaphin clearance of Staphylococcus aureus infection, overview
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
-
169 U/mg (prolysostaphin), 760 U/mg (mature lysostaphin), 1 unit is defined as the amount of enzyme that causes 50% reduction of turbidity in 10 min at 37 C of an inactivated Staphylococcus carnosus TM300 cell suspension
additional information
-
-
assay method with N-acetylhexaglycine as substrate
additional information
-
-
-
additional information
-
-
-
additional information
-
-
-
additional information
-
-
-
additional information
-
-
-
additional information
-
-
depending on the purification step the specific activity varied between 300 U/mg and 1500 U/mg
additional information
-
-
970 U/mg
additional information
-
-
11900 units per mg, after 12 fold purification, 1 unit is defined as the amount of enzyme required to decrease the OD(A600) of a suspension of Staphylococcus aureus 237 cells by 0.001 at 37C in 5 min in a 3 ml reaction volume
additional information
-
-
in vitro activity of recombinant lysostaphin against Staphylococcus aureus isolates from hospitals in Beijing, China, with MIC values of 30-2000 ng/ml, overview
additional information
-
-
activities of three enzymes namely, glycylglycine endopeptidase, endo-beta-N-acetyl glucosamidase and N-acteyl muramyl-L-alanine amidase. Lysostaphin rapidly lyses actively growing and non-dividing cells including staphylococci in biofilms. Glycylglycine endopeptidase lyses staphylococcal cells by hydrolyzing glycylglycine bonds in the polyglycine bridges which form cross links between glycopeptide chains in the cell wall peptidoglycan of Staphylococcus aureus cells. Lytic principle of lysostaphin is a peptidase which liberates N-terminal glycine and alanine from Staphylococcus aureus cell wall. Hexosaminidase is present in lysostaphin preparation is specific for the gluocainyl-muramic acid bond of the bacterial carbohydrate backbone
additional information
-
-
lysostaphin inhibits growth of methicillin-resistant Staphylococcus aureus USA300 at a concentration of 0.096 mg/ml. Growth inhibition of the two enzymes lysostaphin and LysK in combination is greater than either enzyme alone
additional information
-
-
treatment of methicillin-susceptible Staphylococcus aureus strain ATCC 25923 with lysostaphin results in major structural changes in the form of cell swelling, splitting of the septum, creation of nanoscale perforations, decrease of the bacterial spring constant and cell wall stiffness (atomic force microscopy)
additional information
-
-
gene product 13 is a protein at the tail tip of the Bacillus subtilis bacteriophage phi29 with D,D-endopeptidase functionality
additional information
-
-
FVB albino dams infected with methicillin-resistant Staphylococcus aureus for analysis of lysostaphin impact on pharmacokinetics, minimum inhibitory concentration(0.008 microg/ml) and minimum bactericidal concentration (0.015 microg/ml)
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4
-
-
half-life of the staphylolytic activity approximately 2 months
6
-
-
highest specific activity (turbidity reduction assay)
7.4
-
-
assay at
7.5
-
-
enzymeproduced with pBAD vector system
7.5
-
-
in vivo assay at
8
-
-
enzyme obtained from pET vector system
additional information
-
-
the pH value near a negatively charged cell is supposed to be strongly shifted to acidity as compared to the pH of the solution volume. This shifts the enzyme pH dependence curve in solution to alkalinity, overview
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6
9
-
strong activity (turbidity reduction assay)
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5
-
-
half-life of the staphylolytic activity approximately 2 months
37
-
-
in vivo assay at
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
37
-
-
stable for 72 h, recombinant enzyme
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
9.5
-
-
calculated from amino acid sequence
9.7
-
-
mature enzyme
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
the enzyme is secreted
-
Manually annotated by BRENDA team
Staphylococcus capitis EPK1, Staphylococcus simulans NRRL B-2628
-
-
-
-
Manually annotated by BRENDA team
Staphylococcus simulans TNK1
-
the enzyme is secreted
-
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
Staphylococcus aureus (strain NCTC 8325)
Staphylococcus aureus (strain NCTC 8325)
Staphylococcus aureus (strain NCTC 8325)
Staphylococcus aureus (strain NCTC 8325)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
16260
-
-
ESI-TOF MS, endopeptidase domain (residues 1-140)
18400
-
-
N-terminal domain of gene product 13, 159 amino acids, calculated mass
22300
-
-
C-terminal domain of gene product 13, 202 amino acids, calculated mass
24500
27800
-
sedimentation equilibrium, sedimentation and diffusion, gel filtration, polyacylamide electrophoresis
26800
-
-
246 amino acids, mature peptide, recombinant enzyme
26900
-
-
calculated from the plot of mobility versus log molecular weight of standards
26920
-
-
mature lysostaphin, calculation from nucleotide sequence
27000
-
-
mature enzyme
27000
-
-
SDS-PAGE, mature enzyme
27200
-
-
theoretically estimated value
28010
-
-
ESI-TOF MS, residues 247-493
29000
-
-
Sephacryl S-200 gel filtration, His6-fused lysostaphin
35600
-
-
mature enzyme, calculation from nucleotide sequence
39310
-
-
proenzyme, calculation from nucleotide sequence
40900
-
-
gp13 wild-type, 365 amino acids, calculated mass
42000
-
-
preproenzyme
42210
-
-
preprolysostaphin, calculation from nucleotide sequence
48000
-
-
SDS-PAGE in presence and absence of reducing agents
49120
-
-
prolysostaphin, calculation from nucleotide sequence. Prolysostaphin is processed to the mature enzyme by an extracellular cysteine protease
64000
68000
-
SDS-PAGE of prolysostaphin; the overestimated value is due to a high glutamate content. Prolysostaphin is processed to the mature enzyme by an extracellular cysteine protease
64000
68000
-
SDS-PAGE of prolysostaphin
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
heterodimer
-
C-terminal and N-terminal domain
monomer
-
1 * 64000-68000, prolysostaphin, the overestimated value is due to a high glutamate content
monomer
-
1 * 27000, calculated from amino acid sequence
monomer
Staphylococcus simulans NRRL B-2628
-
1 * 64000-68000, prolysostaphin, the overestimated value is due to a high glutamate content
-
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
proteolytic modification
-
the preprolysostaphin is proteolytically processed to the mature enzyme
proteolytic modification
-
lysostaphin is secreted as a proenzyme. Proteolytic cleavage of 13 N-terminal tandem repeats generates mature lysostaphin with 2 functionally separable domains
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
Computational modeling of the gene product 13-muropeptide complex for analysing the orientation of the scissile bond
-
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4
-
-
half-life at 5C 2 months
5.5
-
-
7% of activity is observed at pH of 5.5
5.5
-
-
7% of activity is observed at pH 5.5
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5
-
-
half-life at pH 4 is 2 months
20
30
-
stable for 24 hours at pH 8.0
40
-
-
stable for 4h with 80% activity
50
-
-
15 min, slight decrease of activity
50
-
-
when incubated at temperatures from 25 to 70C the enzyme is stable up to 50C, above the activity is rapidly lost
60
-
-
15 min, marked decrease of activity
75
-
-
15 min, complete inactivation
100
-
-
15 min, stable. Activity is destroyed after 30 min
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
lysostaphin is relatively stable when conjugated with polyethylene glycol and it maintains its activity in human serum
-
relatively unaffected by salt concentrations between 200 and 500 mM, LysK has a higher specific activity than lysostaphin at NaCl concentrations higher than 150 mM (turbidity reduction assay)
-
trypsin inactivates
-
ORGANIC SOLVENT
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
n-Butanol
-
stable to n-butanol-acetic acid-water, 3:1:1 v/v
n-Butanol
Staphylococcus sp. K-6-WI
-
stable to n-butanol-acetic acid-water, 3:1:1 v/v
-
Pyridine
-
stable to pyridine-water, 65:35 v/v
Pyridine
Staphylococcus sp. K-6-WI
-
stable to pyridine-water, 65:35 v/v
-
tert-Butanol
-
stable to tert-butanol-acetic acid-water, 74:15:25 v/v
tert-Butanol
Staphylococcus sp. K-6-WI
-
stable to tert-butanol-acetic acid-water, 74:15:25 v/v
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
5C, 1 mM acetate buffer pH 4, half-life 2 months
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
ammonium sulfate precipitation and ion exchange chromatography
-
cobalt Sepharose column chromatography
-
from commercial preparation
-
immobilized metal affinity chromatography on Ni-NTA resin
-
lysostaphin can be purified from Staphylococcus simulans strains
-
metal-affinity chromatography and cation exchange chromatography
-
method not mentioned
-
recombinant enzyme from Escherichia coli
-
recombinant His-tagged lysostaphin comprising residues 1 to 154 from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
-
recombinant lysostaphin from Escherichia coli to homogeneity
-
zinc metal-chelate chromatography, cation-exchange chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expressed in Escherichia coli and Lactococcus lactis
-
identification of host factors in Lactococcus lactis subspecies cremoris MG1363 that affect the secretion efficiency of the peptidoglycan hydrolase lysostaphin by isolation and characterization of mutants generated by random transposon mutagenesis that had altered lysostaphin activity. Mutants with higher lysostaphin activity contain insertions in 4 different genes
-
overexpression in Escherichia coli
-
amplified from pRG5 recombinant plasmid and cloned into Escherichia coli expression pTYB12 vector which allows the overexpression of a target protein as a fusion to a self-cleavabe affinity tag
-
expressed in Escherichia coli
-
expressed in Escherichia coli BL21(DE3) cells
-
expressed in Escherichia coli strain BL21(DE3)
-
expressed in HeLa cells
-
expression in Bacillus sphaericus
-
expression in Escherichia coli
-
expression in Escherichia coli with pUC19 as vector
-
expression in Staphylococcus simulans, Staphylococcus aureus
-
expression of His-tagged lysostaphin comprising residues 1 to 154 in Escherichia coli strain BL21(DE3)
-
expression of the functional mature enzyme fused to secretion signals of the Sep protein from Lactobacillus fermentum BR11, to direct the enzyme secretion from recombinant cells, in Lactobacillus fermentum strain BR11, Lactobacillus rhamnosus strain GG, Lactobacillus plantarum ATCC 14917 and Lactococcus lactis strain MG1363, method development and production levels, overview
-
recombinant expression in Escherichia coli
-
recominant expression of His-tagged lysostaphin by Escherichia coli BL21 (DE3) pEA3
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
D195A
-
mutant of gene product 13, reduced general protein folding and Zn2+ binding (circular dichroism and particle-induced X-ray emission spectroscopy)
D195C
-
mutant of gene product 13, general protein folding and Zn2+ binding are maintained (circular dichroism and particle-induced X-ray emission spectroscopy)
D37A
-
no endopeptidase activity
H114A
-
no endopeptidase activity
H116A
-
no endopeptidase activity
H33A
-
no endopeptidase activity
H83A
-
no endopeptidase activity
T107A
-
reduced endopeptidase activity
Y81A
-
reduced endopeptidase activity
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
analysis
-
improvement of lipid extraction of staphylococcal cells
diagnostics
-
detection of Staphylococcus aureus
food industry
-
antistaphylococcal agent
medicine
-
therapeutic agent for the treatment of Staphylococcus aureus infections
medicine
-
staphylococcal disease pathogenesis
medicine
-
treatment of staphylococcal infections
medicine
-
application as effective antibiotic drug against infections with Staphylococcus aureus
medicine
-
lysostaphin is useful in treatment of neonatal Staphylococcus aureus infection, overview
medicine
-
potential therapeutic agent against antibiotic-resistant Staphylococcus aureus infections
medicine
-
lysostaphin, in combinantion with the antibiotic ranalexin, is effective against wound or systemic infections caused by meticillin-resistant Staphylococcus aureus. The combination is significantly more effective than treatment with ranalexin or lysostaphin alone. Ranalexin and lysostaphin can be incorporated in wound dressings for the prevention and treatment of topical Staphylococcus aureus infections
medicine
-
lysostaphin is an effective treatment as well as prophylaxis for Staphylococcus aureus biofilms on indwelling catheters
medicine
-
lysostaphin can be used potentially as a topical disinfectant or decolonizing agent against methicillin-resistant Staphylococcus aureus infection on on human skin prior to operations. The combination is significantly more effective than treatment with ranalexin or lysostaphin alone, overview
medicine
-
lysostaphin has potential for use as an antistaphylococcal agent for treatment of infections caused by antibiotic-resistant strains of Staphylococcus aureus
medicine
-
lysostaphin-functionalized cellulose fibers show activity against Staphylococcus aureus in an in vitro skin model (HaCaT keratinocyte) with low toxicity toward keratinocytes, suggesting good biocompatibility for these materials as antimicrobial matrices in wound healing applications
medicine
-
lysostaphin can be used in combination with clarithromycin for the eradication of Staphylococcus aureus biofilms with a minimal biofilm eradication concentration reduction of 9 and 6fold dilutions on meticillin-resistant S. aureus and meticillin-susceptible S. aureus, respectively
pharmacology
-
lysostaphin is useful for treatment for systemic Staphylococcus aureus infection in a mouse model
medicine
-
lysostaphin is an antistaphylococcal agent. The enzyme is efficacious in the treatment of bovine mastitis caused by staphylococci
additional information
-
antistaphylococcal agent in clinical laboratories