phiBIOTICS

Name	Lysostaphin
Recommended Name ?	lysostaphin
Alternative Name ?	bacteriocin glycyl-glycine endopeptidase metalloendoprotease metalloendoproteinase
Uniprot ID	P10547
General Mode of Action	Enzymatic cleavage of peptidoglycan which results in a rapid lysis of the bacterial cell.
phiBIOTICS Family ?	Metallopeptidase
Reaction ?	1. Hydrolysis of the -Gly-\|-Gly- bond in the pentaglycine inter-peptide link joining staphylococcal cell wall peptidoglycans EC: 3.4.24.75 Corresponding Pfam domain: Peptidase_M23 Evidence: experimental (PubMed: 14317407)
Source Organism ?	Staphylococcus simulans
Target Organism ?	Staphylococcus aureus
Note on Target	Lysostaphin also target coagulase-negative staphylococci.
Disease ?	Infective endocarditis Nosocomial infections Osteomyelitis Pneumonia
State ?	Tested
Reference ?	3547405 14317407 18607587

Studies found: 31

Antimicrobial Agent	Study Type	Model	Administration	Relevant Results	Adverse Effects and Other Issues	Reference ?
Lysostaphin	in vitro	bovine mammary gland epithelial cells	adenoviral-vector	Efficient transduction of modified gene resulted in secretion of functional Gln^125,232 lysostaphin (0.8 µg/mL) into cell culture medium. Biological activity was approximately at level 20% of native recombinant-lysostaphin produced by bacteria.		12201521
Lysostaphin	in vitro	set of 257 Staphylococcus aureus clinical isolates (colected in Bejing, China) including 168 MSSA strains and 89 MRSA strains		Efficient lytic activity against all 257 staphylococcal clinical isolates tested. Lysis of MSSA and MRSA strains was equally effective. Activity of lysostaphin was sustained for at least 6 h.		17172520
Lysostaphin	in vitro	set of 429 Staphylococcus aureus clinical isolates (colectes in Germany) including 23 MRSA strains and low level (3.5-9.1%) ORSA strains		Efficient lytic activity against all 429 staphylococcal isolates tested (isolates obtained from blood of individuals with S. aureus bacteremia or from nasal swabs of S. aureus carriers).		14576128
Lysostaphin	in vitro	catheters colonized with several strains of Staphylococcus aureus		Lysostaphin absorbed onto surface of catheters (catheters coated with 0.1 mg/mL of enzyme for 1 h and then washed extensively with 50 mL of PBS) efficiently prevented catheter colonization and moreover eradicated bacteria in solution within the catheter lumen.	Other Issues Killing efficacy increased with coating time of catheters. Significant effect was achieved upon 5 min of coating with enzyme, complete clearence occured after 15 min of coating. Enzyme retained significant antibacterial activity for at least 4 days. Incubation of catheters with human serum for 24 h did not decreased enzyme activity, thus human proteins shown no deterioration of enzyme activity.	15215130
Lysostaphin	in vivo	cotton rat model of nasal Staphylococcus aureus colonization	intranasal	Lysostaphin (110 µg in 10 µL of PBS) administered once daily for 2 days in a dropwise fashion several days after intranasal instillation with MRSA eliminated nasal colonization in 3 of 9 treated animals.	Resistance To adress an issue of lysostaphin resistance, 91 S. aureus strains from 22 nares of treated rats were tested. No intranasal lysostaphin-resistant colonies were found.	12709327
Lysostaphin	in vivo	rabbit model of experimental VISA aortic valve endocarditis	intravenous	A single dose of lysostaphin (100 mg/kg of body weight) followed by 3-day free drug period had mean vegetation bacterial count reduction of 7.27 and 6.63 log¹⁰ CFU/g (compared to the control or VAN treated animals respectively); sterilized 43% of aortic valve vegetation. Enzyme dose (30 mg/kg) administered twice daily in 3 days had mean vegetation bacterial count reduction of 8.27 and 7.63 log¹⁰ CFU/g (compared to the control or VAN treated animals respectively); sterilized of aortic valve vegetations. Rates were significantly higher for either treatment with enzyme than for animals treated with VAN.		10390235
Lysostaphin	in vivo	rat model of neonatal Staphylococcus aureus infection	intraperitoneal	Lysostaphin (1 mg/kg of body weight) administered 30 min or 6 h after inoculation with MSSA strain resulted in significantly improved survival of suckling rats, compared to the control animals. Efficiency of treatment was comparable to those of VAN or OXA.	Health Effect Treatment with lysostaphin or other two antibiotics did not affect weight gain of pups.	17420212
Lysostaphin	in vivo	rabbit model of experimental MRSA endophthalamitis	intravitreal	Lysostaphin (0.1 mg/mL) administered 8 or 24 h after inoculation with 200 CFU or 50 CFU of Staphylococcus aureus respectively, significantly decreased bacterial counts in the vitreous by approximately 6 logs. Moreover treatment 8 h postinfection sterilized 88% of eyes; 24 h postinfection 50% of eyes compared to the untreated eyes.	Other Issues Only treatment 8 h postinfection was able to prevent the development of pathogenic inflammatory changes in the eyes.	11584345
Lysostaphin	in vivo	rabbit model of MRSA or MSSA keratitis	topical	Lysostaphin (2.8 mg/mL) administered in dropwise fashion every 30 min from 4 to 9 h or 10 to 15 h postinfection. Early treatment resulted in 100% sterilization of the corneas, whereas treatment with VAN failed to sterilize any cornea. Efficiency of late lysostaphin treatment was higher than that of VAN.	Other Issues Studies with Styphylococcus aureus mutants lacking erosion of cornea shown ability of lysostaphin to penetrate the intact cornea and eradicate bacteria.	10798659
Lysostaphin	in vivo	rabbit model of MRSA keratitis and endophthalamitis	topical	Lysostaphin was sufficient to cure infection in immunized animals. In keratitis experimental model a single dose of lysostaphin (3 mg/mL) administered in dropwise fashion 10 to 15 h postinfection (inoculation with 100 CFU of MRSA) every 30 min, resulted in reduction to less then 1 log CFU in occular tissue compared to the untreated animals with 10x6 log CFU/cornea. In endophthalamitis experiments a single dose of lysostaphin (1 mg/mL) administered 8 h postinfection (50 CFU of MRSA) efficiently sterilized all eyes of animals.	Toxicity Animals were immunized against lysostaphin by subcutaneous, intranasal and intraocular routes resulting in high serum antibody titers, highest titers were observed after subcutaneous immunization. Despite presence of anti-lysostaphin antibodies, lysostaphin treatment was effective, no adverse reaction or deterioration of its activity were noted.	12454041
Lysostaphin	in vivo	cotton rat model of nasal Staphylococcus aureus colonization	intranasal	Three doses of 0,5% (actual dose, ~150 µg) lysostaphin in cream administered over 3 days were sufficient to completely eliminate nasal colonization in 100% of treated animals. A single dose of 0,5% cream eradicated colonization in 93% of the treated animals. Decrescent doses of enzyme in cream resulted in lower eradication rates: 3 doses of 0,125% (actual dose, ~37.5 µg) lysostaphin in cream eliminated nasal colonization in 55% of the animals, a single dose only in 20% of the animals.	Resistance To adress an issue of lysostaphin resistance, 91 S. aureus strains from 22 nares of treated rats were tested. No intranasal lysostaphin-resistant colonies were found. Other Issues Despite fact that antibacterial acitivty of lysostaphin was maintained for at least 24 h post instillation, reducing efficiency of enzyme could be caused by rapid clearance from the surface of the mucose membrane. To extend the mucosa residence time, two different delivery systems were developed, hydrophilic cream formulation and mucoadhesive formulation of the enzyme.	12709327 15553221
Lysostaphin	in vivo	murine model of MRSA neonatal sepsis	intraperitoneal	Lysostaphin (10mg/kg or 15mg/kg of body weight per dose) administered after inoculation with lethal dose of Staphylococcus aureus rescued 41% or 52% mice (depending on the dose of enzyme) compared to the VAN treated animals (34%) and untreated control (6,2%) respectively.	Health Effect No significant difference in growth among groups of pups were noted.	19127212
Lysostaphin	in vivo	caprine non-lacting mammary gland cells	intramammary infusion	Concentration levels of enzyme in mammary secretions ranged fom 0.2 to 0.6 mg/mL on day one postinfusion and 0.9 to 1.1 mg/mL on day two. The enzyme concentrations were fairly low, resulting in deficient staphylolytic activity.	Toxicity Strong humoral immune response to both the adenoviral vector and enyzyme was observed. Lysostaphin reactive antibodies (mainly IgG subclass) were detected from 11th day and remainded at high level up to 42 days. Adenoviral antibodies were detected within seven days after infusion with dramatic increase in their titers.	12201521 15202644
Lysostaphin	in vivo	transgenic mice	transgenesis	Transgenic mice efficiently secreting lysostaphin into their milk, were divided in three lines with concentration ranged from ~~0.06, ~0.13 and ~0.8 mg/mL, respectively. All glands of mice from highly expressing line (0,8 mg/mL) were completely resistant to staphylococcal infection, as were 40% of glands from other two lines compared to the 77% infected glands of the non-transgenic mice.	Resistance Degree of resistance to bacterial infection correlated with the enzyme content in glands. Health Effect In transgenic glands with viable count of staphylococci, no signs of inflammation or histopathology effects were noted. Secretion of lysostaphin were restricted essentially to cells of transgenic mice mammary glands, did not affect structure of glands, quality of milk and have no effect on milk proteins. Other Issues Antibacterial acitivity of enzyme was 5 to 10 times lower than that of native derived recombinant enzyme.	11135555
Lysostaphin	in vivo	transgenic cow	transgenesis	Concentrations of enzyme ranged from 0.9 to 14 mg/mL and remained constant during most of lactation. Following intramammary infusion of Staphylococcus aureus 14% of transgenic glands became infected compared to the 71% of non-transgenic cow glands.	Resistance Degree of resistance to bacterial infection correlated with the enzyme content in glands. Health Effect Secretion of the enzyme did not affect quality and volume of produced milk or the physiology of the trangenic animals. In infected glands, the severity and duration of infections were less in non-transgenic animals. Other Issues Antibacterial activity was 15% of the biological activity of the bacteria derived recombinant enzyme.	15806099
Lysostaphin	in vivo	bovine model of Staphylococcus aureus mastitis	intramammary infusion	Minimal effective dose of recombinant-lysostaphin was found to be 100 mg. The cure rate in animals that recieved minimal dose of enzyme dissolved in PBS postinfection was 20% compared to the 29% for sodium cepharin in PBS and 57% for its commercial formulation, respectively.	Health Effect Expresion of enzyme in mammary glands did not affect health of animals. Other Issues It was found that lysostaphin is fairy stable, incubation of the enzyme in milk at 37°C for 72 h did not lead to deterioration of its antibacterial acitivity. Despite the maintance of antibacterial activity of enzyme up to 120 h, owerall effectivness of this therapeutic regimen was decreased by relatively quick relapses (within 5 to 6 milkings), likely due to bacteria internalization by cells of the mammary glands, whereby contributes to the reinfections (lysostaphin cannot kill intracellular bacteria).	1787188 16702252
Lysostaphin	in vivo	rabbit model of experimental coagulase-negative Staphylococcus endophthalamitis		Lysostaphin (250 µg) administered 8 h postinfection significantly reduced bacterial counts for most staphylococci strains tested, including decrease by 6 logs for MRSE; by 2 logs for Staphylococcus werneri and for other strains a 4 log reduction compared to the untreated eyes. No reduction were observed for Staphylococcus haemolyticus, 2 MRSE strains and 1 Staphylococcus cohni strain.	Other Issues A difference between the MIC values for some strains and their susceptibility to lysostaphin in vivo was found.	16531279
Lysostaphin + Gentamicin	in vitro	set of 98 MRSA clinical isolates derivated from blood, nares, tissue, sputum, urine and from other sites		Effective synergistic inhibition was noted for lysostaphin and GEN combinations (difference of 2,898 + 0,13 log¹⁰ CFU/mL).		17141452
Lysostaphin + Vancomycin	in vivo	rabbit model of experimental MRSA aortic valve endocarditis	intravenous	Lysostaphin (5 mg/kg of body weight) administered every 8 h sterilized vegetation in 10 of 11 treated animals and had mean reduction in aortic valve vegetation bacterial counts by 8.5 log¹⁰ CFU/g. A single dose of lysostaphin was less effective in reducing vegetation bacterial counts density only by 3.5 log¹⁰ CFU/g. VAN (30mg/kg) given every 12 h did not sterilized vegetations in animals and reduced staphylococcal counts by only 4.8 log¹⁰ CFU/g of tissue . Combination of lysostaphin (5 mg/kg) given once daily and VAN (30 mg/kg) given twice daily resulted in reducing mean vegetation bacterial density by 7.5 log¹⁰ CFU/g, therapeutic effect was significantly better than that for either agent alone.	Resistance To adress immunological response issues, rabitts recieved weekly intravenous lysostaphin treatment for over 9 weeks. No signs of adverse effects were observed.	9624475
Lysostaphin + Lysozyme	in vitro	set of 235 Staphylococcus strains including 84 Staphylococcus aureus strains and 151 strains belonging to 9 different coagulase-negative staphyloccal species		Lysozyme considerably reduced lysostaphin MIC-values in all 235 strains tested with the reduction ranged from 16- to 200-fold. In particular, 200-fold decrease was noted for Staphylococcus cohni, 16- to 45-fold decrease was observed for other strains tested, using combination (4 µg lysostaphin plus 5 mg lysozyme per mL).		7081976
Lysostaphin + LysK	in vitro	MRSA USA300 strain		Combination of lysostaphin and LysK resulted in effective synergistic inhibition. The two most effective combinations were 0.027 and 5.71 µg/mL for lysostaphin and 0.018 and 11.4 µg/mL for LysK, representing concentrations that correspond to 30%, 18% and 16%, 33%, respectively, of each agents MICs.	Other Issues Both antimicrobial agents shown strong activity over a broad pH range (from pH 6 to 9) with their activity near maximal at physiological pH. LysK was found to have higher specific activity than lysostaphin at NaCl concentrations >150 mM.	18721148
Lysostaphin + Ranalexin + Magainin II + Dermaseptin S3(1-16)	in vitro	set of various bacterial strains including: MRSA252 MSSA476 ESBL Escherichia coli VR Enterococcus faecium VR Enterococcus faecalis Streptococcus sanguis		Combination of lysostaphin with RNL resulted in effective synergisitc inhibition, specific against MRSA and MSSA strains. No activity was shown against other Gram-positive and negative bacterial strains tested. Synergistic effects were also observed for combinations of lysostaphin with other two CAMPs: MG II and DS-S3(1-16).		17324959
Lysostaphin + Ranalexin	in vivo	rabbit model of wound infection	topical	Combination of lysostaphin (0.0045 mg) with RNL (0.225 mg) resulted in synergistic growth inhibition, administratrated (by moistening the gauze of fresh Hilltop chambers) 24, 48 and 72 h post-inoculation with 10x9 CFU of MRSA strain reduced viable titers in the wound by ca. 3.5 log¹⁰ CFU compared to the untreated animals. Combined treatment was shown to be more effective than administration of either antibacterial agent alone.	Toxicity Combined treatment was proved to be non-toxic by haemolysis and Vero cell viability assays, compounds shown signs of toxicity to erythrocytes and Vero cells when used in concentrations greater than those required for their antibacterial activities.	20206480
Lysostaphin + Ranalexin	in vivo	mice model of systemic MRSA infection	intravenous	Combination of lysostaphin (0.1 mg/kg) with RNL (12 mg/kg of body weight) resulted in synergistic growth inhibition, administratrated 2; 26; and 50 h post-inoculation with 1.7x 10x7 CFU of MRSA strain reduced staphylococcal kidney burden by ca. 1.5 log¹⁰ CFU compared to the untreated animals. Combined treatment was shown to be more effective than administration of either antibacterial agent alone.	Toxicity Combined treatment was proved to be non-toxic by haemolysis and Vero cell viability assays, compounds shown signs of toxicity to erythrocytes and Vero cells when used in concentrations greater than those required for their antibacterial activities.	20206480
Lysostaphin + β-lactam antibiotics + Bacitracin + Polymyxin B	in vitro	set of Staphylococcus aureus strains including: MRSA (RN451;RN1753) MSSA (W559;W562;Q877)		Effective synergistic inhibition demonstrated for lysostaphin combinations with BLA (including benzylpenicillin, methicillin, cephalosporin B) BAC and PMB against both MRSA, MSSA strains tested.	Other Issues PMB shown no activity in milk.	8112040
Lysostaphin + Nafcillin	in vivo	rabbit model of experimental ORSE aortic valve endocarditis	intramuscular intravenous	Combination of lysostaphin (1 mg/kg of body weight) with NAF (200 mg/kg) administered intravenously and intramuscularly twice a day respectively resuleted in effective synergistic growth inhibition against ORSE, reducing bacterial counts by 5.32 log¹⁰ CFU/g. Combined treatment was shown to be more effective than administration of either antibacterial agent alone.		12019130
Lysostaphin + Nafcillin	in vivo	rabbit model of experimental ORSA aortic valve endocarditis	intramuscular intravenous	Combination of lysostaphin (1mg/kg of body weight) with NAF (200 mg/kg) administered intravenously and intramuscularly twice a day respectively, resulted in effective synergistic growth inhibition against ORSA and lead to mean reduction of bacterial counts by 7,5 log¹⁰ CFU/g compared to the controls. Combined treatment was shown to be more effective than administration of NAF (1,35 log¹⁰ CFU/g) or lysostaphin (1,65 log¹⁰ CFU/g) alone.	Resistance Following in vitro exposure to subinhibitory levels of enzyme lysostaphin-resistant mutants emerged (mutation in femA gene). Combined regimen with β-lactame antibiotics can supress resistance.	11302806

Catalogue of Therapeutic Enzybiotics

Lysostaphin

Studies found: 31