A novel secreted metzincin metalloproteinase from <i>Bacillus intermedius</i>

Sabirova, A. R.; Рудакова, Н. Л.; Balaban, N. P.; Ilyinskaya, O.P.; Demidyuk, Ilya V.; Kostrov, Sergey V.; Rudenskaya, G. N.; Шарипова, М. Р.

doi:10.1016/j.febslet.2010.09.049

Cited by 41 publications

(19 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…To verify the stability of Ficin in the culture liquid, the proteolytic activity was measured using azocaseine as substrate37 in wells after the enzyme addition. During the first 4 hours more that 90% of the initial activity was detectable in the liquid (Fig.…”

Section: Resultsmentioning

confidence: 99%

Targeting microbial biofilms using Ficin, a nonspecific plant protease

Baidamshina

Trizna

Holyavka

et al. 2017

Sci Rep

111

View full text Add to dashboard Cite

Biofilms, the communities of surface-attached bacteria embedded into extracellular matrix, are ubiquitous microbial consortia securing the effective resistance of constituent cells to environmental impacts and host immune responses. Biofilm-embedded bacteria are generally inaccessible for antimicrobials, therefore the disruption of biofilm matrix is the potent approach to eradicate microbial biofilms. We demonstrate here the destruction of Staphylococcus aureus and Staphylococcus epidermidis biofilms with Ficin, a nonspecific plant protease. The biofilm thickness decreased two-fold after 24 hours treatment with Ficin at 10 μg/ml and six-fold at 1000 μg/ml concentration. We confirmed the successful destruction of biofilm structures and the significant decrease of non-specific bacterial adhesion to the surfaces after Ficin treatment using confocal laser scanning and atomic force microscopy. Importantly, Ficin treatment enhanced the effects of antibiotics on biofilms-embedded cells via disruption of biofilm matrices. Pre-treatment with Ficin (1000 μg/ml) considerably reduced the concentrations of ciprofloxacin and bezalkonium chloride required to suppress the viable Staphylococci by 3 orders of magnitude. We also demonstrated that Ficin is not cytotoxic towards human breast adenocarcinoma cells (MCF7) and dog adipose derived stem cells. Overall, Ficin is a potent tool for staphylococcal biofilm treatment and fabrication of novel antimicrobial therapeutics for medical and veterinary applications.

show abstract

Section: Resultsmentioning

confidence: 99%

Targeting microbial biofilms using Ficin, a nonspecific plant protease

Baidamshina

Trizna

Holyavka

et al. 2017

Sci Rep

111

View full text Add to dashboard Cite

show abstract

“…We have shown that the bacterium B. pumilus 3 19 secretes into the medium, together with serine proteinases (subtilisin like proteinase and glutamyl endopeptidase), a minor protein (metalloen dopeptidase) [2]. This enzyme can be neither isolated from the culture liquid of the wild type strain nor char acterized due to its extremely low content [3].…”

Section: Introductionmentioning

confidence: 99%

“…The gene sequence of B. pumilus 3 19 extracellular 1 Corresponding author: e mail: Nellybalaban@yandex.ru. metalloendopeptidase has been established previously (AN EU678894) [2]. The gene (mprBp) structure analysis made it possible to ascribe this protein to the clan of metzincins [3].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

The novel ADAMs-like microbial metalloendopeptidase

et al. 2012

View full text Add to dashboard Cite

“…Crude enzyme solution (CES)Partially purified peptidase (PPPase)b)An extracellular metallopeptidase produced by Bacillus amyloliquefaciens FE-K155Trakya Univ J Nat Sci, 21(1):[47][48][49][50][51][52][53][54][55][56][57][58][59][60][61] 2020 Peptidase activity results at different temperatures.…”

mentioning

confidence: 99%

PARTIAL PURIFICATION AND CHARACTERIZATION OF AN EXTRACELLULAR METALLOPEPTIDASE PRODUCED BY Bacillus amyloliquefaciens FE-K1

Erem

İnan

Budak

et al. 2020

Trakya University Journal of Natural Sciences

View full text Add to dashboard Cite

The aim of this study was to purify and characterize the peptidase of Bacillus amyloliquefaciens (Fukumoto) (strain FE-K1) isolated from ropey bread. Peptidases were purified from crude enzyme solution by affinity chromatography with an efficiency of 25 % and a purification coefficient of 1.53. The optimum pH of partially purified peptidase (PPPase) solution was determined as 7.5 and the peptidases retained approximately 90 % of their initial activity in the pH range 7.0-8.5 following incubation at 37°C for 2 h. The optimum temperature for the PPPase was 60°C. The approximate molecular weight of the PPPase was determined as 36 kDa. Inactivation of the PPPase in the presence of O-FEN and EDTA showed them to be metallopeptidases and 5 mM of K +1 and 5 mM of Mn +2 ions increased the enzyme activity by 4 % and 6.15 %, respectively. The presence of Hg +2 , Fe +3 and SDS (0.1-1.0 % w/v) caused inactivation whereas the enzyme retained most of its activity in the presence of 0.1-1.0 % (v/v) Triton X-100, Tween 20 and Tween 80 and 1-20 % (v/v) xylene, ethanol, acetone and acetonitrile. Characterization of the PPPase revealed the enzyme as a neutral serine metallopeptidase compatible with some organic solvents and surfactants.Özet: Bu çalışmanın amacı, sünmüş ekmeklerden izole edilen Bacillus amyloliquefaciens (Fukumoto) (suş FE-K1) ile elde edilen peptidazı saflaştırmak ve karakterize etmektir. Peptidazlar ham enzim çözeltisinden afinite kromatografisi ile % 25 verim ve 1,53 saflaştırma katsayısı ile saflaştırılmıştır. Kısmi olarak saflaştırılmış peptidaz (PPPaz) çözeltisinin optimum pH değeri 7,5 olarak tespit edilmiş olup, pH 7,5-8,0 aralığında peptidaz, 37°C'de 2 saat inkübasyonun ardından başlangıç aktivitesini yaklaşık % 90 oranında korumuştur. PPPaz'ın optimum sıcaklığı 60°C'dir. PPPaz'ın yaklaşık molekül ağırlığı 36 kDa olarak belirlenmiştir. PPPaz'ın O-FEN ve EDTA varlığında inaktive olması, enzimin metallopeptidaz olduğunu göstermiştir. Ayrıca 5 mM K +1 ve 5 mM Mn +2 , enzimin aktivitesini sırasıyla % 4 ve % 6,15 oranında artırmıştır. Hg +2 , Fe +3 ve SDS (% 0,1-1,0 w/v) varlığı enzimin inaktivasyonuna neden olurken, % 0,1-1,0 (v/v) Triton X-100, Tween 20 ve Tween 80; ve % 1-20 (v/v) ksilen, etanol, aseton ve asetonitril varlığında enzim aktivitesini büyük ölçüde korumuştur. PPPaz'ın karakterizasyonu, enzimin bazı organik çözücü ve yüzey aktif maddelerle ile uyumlu nötr bir serin metalopeptidaz olduğunu ortaya çıkarmıştır.

show abstract

A novel secreted metzincin metalloproteinase from Bacillus intermedius

Cited by 41 publications

References 32 publications

Targeting microbial biofilms using Ficin, a nonspecific plant protease

Targeting microbial biofilms using Ficin, a nonspecific plant protease

The novel ADAMs-like microbial metalloendopeptidase

PARTIAL PURIFICATION AND CHARACTERIZATION OF AN EXTRACELLULAR METALLOPEPTIDASE PRODUCED BY Bacillus amyloliquefaciens FE-K1

Contact Info

Product

Resources

About