Topic
Bacillolysin
About: Bacillolysin is a research topic. Over the lifetime, 5 publications have been published within this topic receiving 125 citations.
Papers
TL;DR: The results increase the veterinary and medical importance of the allergens detected in feces and the strong symbiotic association of T. putrescentiae with B. cereus has to coexist with the mite in balance to be beneficial for the mites.
Abstract: Tyrophagus putrescentiae (Schrank, 1781) is an emerging source of allergens in stored products and homes. Feces proteases are the major allergens of astigmatid mites (Acari: Acaridida). In addition, the mites are carriers of microorganisms and microbial adjuvant compounds that stimulate innate signaling pathways. We sought to analyze the mite feces proteome, proteolytic activities and mite-bacterial interaction in dry dog food. Proteomic methods comprising enzymatic and zymographic analysis of proteases and 2D-E-MS/MS were performed. The highest protease activity was assigned to trypsin-like proteases; lower activity was assigned to chymotrypsin-like proteases, and the cysteine protease cathepsin B-like had very low activity. The 2D-E-MS/MS proteomic analysis identified mite trypsin allergen Tyr p3, fatty acid-binding protein Tyr p13 and putative mite allergens ferritin (Grp 30) and (poly)ubiquitins. Tyr p3 was detected at different positions of the 2D-E. It indicates presence of zymogen at basic pI, and mature-enzyme form and enzyme fragment at acidic pI. Bacillolysins (neutral and alkaline proteases) of Bacillus cereus symbiont can contribute to the protease activity of the mite extract. The bacterial exo-chitinases likely contribute to degradation of mite exuviae, mite bodies or food boluses consisting of chitin, including the peritrophic membrane. Thus, the chitinases disrupt the feces and facilitate release of the allergens. B. cereus was isolated and identified based on amplification and sequencing of 16S rRNA and motB genes. B. cereus was added into high-fat, high-protein (dry dog food) and low-fat, low-protein (flour) diets to 1% and 5% (w/w), and the diets palatability was evaluated in 21-day population growth test. The supplementation of diet with B. cereus significantly suppressed population growth and the suppressive effect was higher in the high-fat, high-protein diet than in the low-fat, low-protein food. Thus, B. cereus has to coexist with the mite in balance to be beneficial for the mite. The mite-B. cereus symbiosis can be beneficial-suppressive at some level. The results increase the veterinary and medical importance of the allergens detected in feces. The B. cereus enzymes/toxins are important components of mite allergens. The strong symbiotic association of T. putrescentiae with B. cereus in dry dog food was indicated.
62 citations
TL;DR: An extracellular calcium-dependent protease from a newly isolated Bacillus cereus SV1 was purified, characterized and the gene was isolated and sequenced, suggesting that the purified enzyme is a metalloprotease.
27 citations
Journal Article•
TL;DR: A single fibrinolytic enzyme from Bacillus pseudomycoides B-60 was obtained and its N-terminal sequence was determined to be VTGTNAVGTGKGVLG, which provided a basis for further study of new thrombolytic drugs.
Abstract: OBJECTIVE To purify a single fibrinolytic enzyme from Bacillus pseudomycoides B-60 and to determine its N-terminal sequence and to characterize the fibrinolytic enzyme. METHODS We examined the fibrinolytic enzyme activity by fibrin plate and purified fibrinolytic enzyme by ammonium sulfate fractional precipitation and DEAE anion exchange chromatography. RESULTS Obtained a single protein fraction with fibrinolytic activity (BpFE) from B. pseudomycoides B-60. It appeared as a single band in the SDS-PAGE with a relative molecular weight of 34 kDa. The fibrinolytic activity of the protein was stable at 4-50 degrees C and at pH 5-10. The activity sharply decreased above 50 degrees C, and the total loss of activity at pH 3.0. The enzymatic activity was slightly enhanced by the ions of Ca2+, Mg2+, Mn2+, whereas strongly inhibited by Cu2+ ion. Phenylmethyl sulfonyl fluoride (PMSF) could completely inhibit its activity. In addition, the activity improved when the protein was enzymatically hydrolyzed using trypsin and pepsin. The first 15 amino acids of the N-terminal sequence of the enzyme were determined to be VTGTNAVGTGKGVLG. The partial amino acids sequence alignment study of the enzyme from B-60 strain with bacillolysin, neutral protease and hydrolase which were from B. cereus, B. thuringiensis, B. anthracis and Lactobacillus sp. was carried out, and there is a 100% homogeneity between them. CONCLUSION We obtained a single fibrinolytic enzyme. Through its N-terminal sequence alignment study, a plasmin with high homogeneity to this protein was not found yet. This provided a basis for further study of new thrombolytic drugs.
3 citations
TL;DR: Results indicate that the proteases are natural variants of subtilisin and bacillolysin.
Abstract: Bacillus amyloliquefaciens FSE-68 isolated from meju, a Korean soybean fermentation starter, was identified on the basis of biophysical tests and 16S rRNA gene sequence. A neutral metalloprotease (NPR68) and an alkaline serine-protease (APR68) were purified by ammonium sulfate precipitation and cation exchange chromatography and identified on the basis of their activities at different pH values and the selective protease inhibitors. The molecular weights of NPR68 and APR68 measured with ESI-MS were 32743 (+/- 0.8) and 27443 (+/- 0.5) Da, respectively. Against oxidized insulin chains, the NPR68 has a cleavage preference at the site where leucine is located as a P1' residue followed by phenylalanine, and the APR68 has broad specificity and favors leucine at the P1 site. These results indicate that the proteases are natural variants of subtilisin and bacillolysin.
TL;DR: This eco-friendly treatment based on a new enzyme combination produced by a wild bacterium has potential for meeting the demands of organic wool processing which bans the use of hazardous chemicals and genetic engineering.
Abstract: Bacteria from Patagonian Merino wool were isolated to assess their wool-keratinolytic activity and potential for felt-resist treatments. Strains from Bacillus, Exiguobacterium, Deinococcus, and Micrococcus produced wool-degrading enzymes. Bacillus sp. G51 showed the highest wool-keratinolytic activity. LC-MS/MS analysis revealed that G51 secreted two serine proteases belonging to the peptidase family S8 (MEROPS) and a metalloprotease associated with Bacillolysin, along with other enzymes (γ-glutamyltranspeptidase and dihydrolipoyl dehydrogenases) that could be involved in reduction of keratin disulfide bonds. Optimum pH and temperature of G51 proteolytic activity were 9 and 60 °C, respectively. More than 80% of activity was retained in H2O2, Triton X-100, Tween 20, Lipocol OXO650, Teridol B, and β-mercaptoethanol. Treatment of wool top with G51 enzyme extract caused a decrease in wool felting tendency without significant weight loss (<1.5%). Sparse work has so far been performed to investigate suitable keratinases for the organic wool sector. This eco-friendly treatment based on a new enzyme combination produced by a wild bacterium has potential for meeting the demands of organic wool processing which bans the use of hazardous chemicals and genetic engineering.
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Performance Metrics
| Year | Papers |
|---|---|
| 2017 | 1 |
| 2016 | 1 |
| 2009 | 1 |
| 2008 | 1 |
| 2003 | 1 |