scholarly journals Extracellular proteolytic activities expressed by Bacillus pumilus isolated from endodontic and periodontal lesions

2008 ◽  
Vol 57 (5) ◽  
pp. 643-651 ◽  
Author(s):  
Blair T. Johnson ◽  
Lindsey N. Shaw ◽  
Daniel C. Nelson ◽  
John A. Mayo
Keyword(s):  
Proteolytic Activity ◽  
Bacillus Pumilus ◽  
Serine Proteinase ◽  
Glutamyl Endopeptidase ◽  
Rrna Gene ◽  
Root Canals ◽  
Carbohydrate Fermentation ◽  
Proteolytic Activities ◽  
Marginal Periodontitis

The purpose of the present study was to identify 12 Bacillus isolates that had been obtained from root canals of teeth requiring endodontic therapy and from periodontal pockets in severe marginal periodontitis, and to determine whether these isolates exhibited extracellular proteolytic activity and, using in vitro assays, whether any such activity could degrade substrates that would be pathophysiologically relevant with regard to the production of endodontic and periodontal lesions. Biochemical and carbohydrate fermentation patterns were used in the identification of all strains, which was confirmed by determination of the16S rRNA gene sequence for strain BJ0055. Screening for production of extracellular proteolytic activity by all strains was done with a general proteinase substrate. All isolates were identified as representing Bacillus pumilus and all exhibited extracellular proteolytic activity. The putative pathophysiological relevance of extracellular proteinase production in strain BJ0055 was assessed using fluorophore-labelled elastin and collagen and several chromogenic peptides. Probable classes of proteinases acting on each substrate were investigated using class-specific inhibitors. Activity–pH profiles were determined in buffers at different pH values. Extracellular activities that were caseinolytic, elastinolytic, collagenolytic, glutamyl endopeptidase-like, and alanyl tripeptidyl peptidase-like were observed. No trypsin-like activities were detected. Serine- and chymotrypsin-like serine proteinase activities were detected, with activity observed at neutral and alkaline, but not acidic, pH. B. pumilus strains isolated from endodontic and periodontal lesions exhibited extracellular activities that degrade elastin, collagen and other substrates. These activities may be virulence factors that contribute to tissue damage in apical periodontitis and severe marginal periodontitis.

scholarly journals Proteinases in Excretory-Secretory Products ofToxocara canisSecond-Stage Larvae: Zymography and Modeling Insights

2014 ◽  
Vol 2014 ◽  
pp. 1-9 ◽  
Author(s):  
Gonzalo Ernesto González-Páez ◽  
Fernando Alba-Hurtado ◽  
Carlos Gerardo García-Tovar ◽  
Raúl Argüello-García
Keyword(s):  
Serine Proteinase ◽  
Toxocara Canis ◽  
Serine Proteinases ◽  
Bodily Fluids ◽  
Multiple Components ◽  
Proteolytic Activities ◽  
Secretory Products ◽  
Ph Range ◽  
Physiological Substrates

Components released in excretory-secretory products ofToxocara canislarvae (TES) include phosphatidylethanolamine-binding proteins (TES26), mucins (TES120, MUC2-5), and C-type lectins (TES32, TES70) and their biochemical, immunological, and diagnostic properties have been extensively studied albeit proteinase activities towards physiological substrates are almost unknown. Proteolytic activities in TES samples were first analyzed by gel electrophoresis with gelatin as substrate. Major activities of ~400, 120, and 32 kDa in TES were relatively similar over a broad pH range (5.5–9.0) and all these were of the serine-type as leupeptin abolished gelatinolysis. Further, the ~400 kDa component degraded all physiological substrates tested (laminin, fibronectin, albumin, and goat IgG) and the 120 kDa component degraded albumin and goat IgG while proteinases of lower MW (45, 32, and 26 kDa) only degraded laminin and fibronectin, preferentially at alkaline pH (9.0). By protein modeling approaches using the known sequences of TES components, only TES26 and MUC4 displayed folding patterns significantly related to reference serine proteinases. These data suggest that most of serine proteinase activities secretedin vitroby infective larvae ofT. canishave intriguing nature but otherwise help the parasite to affect multiple components of somatic organs and bodily fluids within the infected host.

Proteolytic effect of Cynara cardunculus rennet for use in the elaboration of ‘Torta del Casar’ cheese

2013 ◽  
Vol 80 (4) ◽  
pp. 429-438 ◽  
Author(s):  
Elena Ordiales ◽  
Maria José Benito ◽  
Alberto Martin ◽  
Margarita Fernández ◽  
Alejandro Hernández ◽  
...  
Keyword(s):  
Proteolytic Activity ◽  
Cynara Cardunculus ◽  
Protein Nitrogen ◽  
Protected Designation Of Origin ◽  
Major Development ◽  
Homogeneous Product ◽  
Proteolytic Activities ◽  
Relationship Of

The purpose of this work was to analyse the influence of rennet from different Cynara cardunculus plants, selected for its clotting and proteolytic activity on caseins, on the characteristics of manufactured ‘Torta del Casar’ cheeses. After classifying the cardoon according to proteolytic activity into five groups of greater or lesser activity, 16 batches of cheeses were made with rennet derived from different wild cardoon plants. We observed a major development of the proteolysis during ripening leading to the generation of non-protein nitrogen compounds. Especially noteworthy was the relationship of amino acid nitrogen (AN) generation with rennet clotting activity after 24 h of maceration, and the fact that the production of biogenic amines was not related to the proteolytic activity of the rennet. The activities of the rennet observed ‘in vitro’ were also developed ‘in vivo’ in the cheeses, with the different rennets used affecting the final sensory characteristics of cheeses. The rennet with high clotting activity after 24 h of maceration was positively correlated with the creaminess, viscosity, and acceptability of the cheese. However, the high proteolytic activity rennet negatively influenced the acidity, bitterness, and creaminess parameters. Therefore the most appropriate cardoons for making this cheese are those with higher clotting activities and moderate proteolytic activities especially on β-casein. The use of controlled and characterised cardoons in the manufacturing process of Torta del Casar is fundamental to obtaining the homogeneous product demanded by the Torta del Casar Registry of the Protected Designation of Origin.

scholarly journals The contribution of individual domains of chloroplast protein AtDeg2 to its chaperone and proteolytic activities

2018 ◽  
Vol 87 (1) ◽  
Author(s):  
Przemysław Jagodzik ◽  
Robert Luciński ◽  
Lucyna Misztal ◽  
Grzegorz Jackowski
Keyword(s):  
Proteolytic Activity ◽  
Thylakoid Membrane ◽  
Linear Structure ◽  
Protein Aggregates ◽  
Pdz Domains ◽  
Protease Domain ◽  
Substrate Protein ◽  
Proteolytic Activities ◽  
First Time

The thylakoid protease AtDeg2 is a non-ATP hydrolyzing chloroplast protease/chaperone peripherally connected with stromal side of thylakoid membrane. Its linear structure consists of protease domain and two PDZ domains. To unveil the significance of individual domains, chaperone and proteolytic activities of AtDeg2, its mutated recombinant versions have been developed and their ability to suppress protein aggregation and resolubilization of protein aggregates as well as the ability to degrade substrate protein was examined in vitro. Our work reveals for the first time that AtDeg2 is able not only to suppress aggregation of denatured proteins, but to resolubilize existing protein aggregates as well. We show that PDZ2 domain contributes significantly to both chaperone and protease activities of AtDeg2, whereas PDZ1 is required for chaperone but superfluous for proteolytic activity. Protease domain – but not S-268 in its catalytic site – contributes to chaperone activities of AtDeg2. These results show an entirely new function of AtDeg2 chaperone/protease (i.e., disaggregation of protein aggregates) and allow to identify structural motifs required for “old” and new functions of AtDeg2.

Effect of the endophytic plant growth promoting Enterobacter ludwigii EB4B on tomato growth

2020 ◽  
Vol 13 (2) ◽  
pp. 54-65 ◽  
Author(s):  
M.E.A. Bendaha ◽  
H.A. Belaouni
Keyword(s):  
Root Length ◽  
Biocontrol Agent ◽  
Growth Promotion ◽  
Dry Weight ◽  
Antagonistic Activity ◽  
Rrna Gene ◽  
Plant Growth Promoting ◽  
Enterobacter Ludwigii ◽  
Tomato Growth

SummaryThis study aims to develop a biocontrol agent against Fusarium oxysporum f.sp. radicis-lycopersici (FORL) in tomato. For this, a set of 23 bacterial endophytic isolates has been screened for their ability to inhibit in vitro the growth of FORL using the dual plate assay. Three isolates with the most sound antagonistic activity to FORL have been qualitatively screened for siderophore production, phosphates solubilization and indolic acetic acid (IAA) synthesis as growth promotion traits. Antagonistic values of the three candidates against FORL were respectively: 51.51 % (EB4B), 51.18 % (EB22K) and 41.40 % (EB2A). Based on 16S rRNA gene sequence analysis, the isolates EB4B and EB22K were closely related to Enterobacter ludwigii EN-119, while the strain EB2A has been assigned to Leclercia adecarboxylata NBRC 102595. The promotion of tomato growth has been assessed in vitro using the strains EB2A, EB4B and EB22K in presence of the phytopathogen FORL. The treatments with the selected isolates increased significantly the root length and dry weight. Best results were observed in isolate EB4B in terms of growth promotion in the absence of FORL, improving 326.60 % of the root length and 142.70 % of plant dry weight if compared with untreated controls. In the presence of FORL, the strain EB4B improved both root length (180.81 %) and plant dry weight (202.15 %). These results encourage further characterization of the observed beneficial effect of Enterobacter sp. EB4B for a possible use as biofertilizer and biocontrol agent against FORL.

scholarly journals The growth response of rice (Oryza sativa L. var. FARO 44) in vitro after inoculation with bacterial isolates from a typical ferruginous ultisol

2021 ◽  
Vol 45 (1) ◽  
Author(s):  
Musa Saheed Ibrahim ◽  
Beckley Ikhajiagbe
Keyword(s):  
16S Rrna ◽  
Bacillus Cereus ◽  
Proteus Mirabilis ◽  
Growth Response ◽  
Gene Sequencing ◽  
16S Rrna Gene Sequencing ◽  
Rrna Gene ◽  
Rice Seedlings ◽  
Rrna Gene Sequencing

Abstract Background Rice forms a significant portion of food consumed in most household worldwide. Rice production has been hampered by soil factors such as ferruginousity which has limited phosphorus availability; an important mineral component for the growth and yield of rice. The presence of phosphate-solubilizing bacteria (PSB) in soils has been reported to enhance phosphate availability. In view of this, the present study employed three bacteria species (BCAC2, EMBF2 and BCAF1) that were previously isolated and proved P solubilization capacities as inocula to investigate the growth response of rice germinants in an in vitro setup. The bacteria isolates were first identified using 16S rRNA gene sequencing and then applied as inoculum. The inolula were prepared in three concentrations (10, 7.5 and 5.0 ml) following McFarland standard. Viable rice (var. FARO 44) seeds were sown in petri dishes and then inoculated with the three inocula at the different concentrations. The setup was studied for 28 days. Results 16S rRNA gene sequencing identified the isolates as: isolate BCAC2= Bacillus cereus strain GGBSU-1, isolate BCAF1= Proteus mirabilis strain TL14-1 and isolate EMBF2= Klebsiella variicola strain AUH-KAM-9. Significant improvement in rice germination, morphology, physiology and biomass parameters in the bacteria-inoculated setups was observed compared to the control. Germination percentage after 4 days was 100 % in the inoculated rice germinants compared to 65% in the control (NiS). Similarly, inoculation with the test isolates enhanced water-use efficiency by over 40%. The rice seedlings inoculated with Bacillus cereus strain GGBSU-1 (BiS) showed no signs of chlorosis and necrosis throughout the study period as against those inoculated with Proteus mirabilis strain TL14-1 (PiS) and Klebsiella variicola strain AUH-KAM-9 (KiS). Significant increase in chlorophyll-a, chlorophyll-b and alpha amylase was observed in the rice seedlings inoculated with BiS as against the NiS. Conclusion Inoculating rice seeds with Bacillus cereus strain GGBSU-1, Proteus mirabilis strain TL14-1 and Klebsiella variicola strain AUH-KAM-9 in an in vitro media significantly improved growth parameters of the test plant. Bacillus cereus strain GGBSU-1 showed higher efficiency due to a more improved growth properties observed.

scholarly journals Chromatographic Characterization and In Vitro Bioactivity Evaluation of Lactobacillus helveticus Hydrolysates upon Fermentation of Different Substrates

2021 ◽  
Vol 11 (2) ◽  
pp. 811
Author(s):  
Federica Ianni ◽  
Alessandra Anna Altomare ◽  
Beniamino T. Cenci-Goga ◽  
Francesca Blasi ◽  
Luca Grispoldi ◽  
...  
Keyword(s):  
Liquid Chromatography ◽  
Proteolytic Activity ◽  
Bioactive Peptides ◽  
Biological Activities ◽  
Skim Milk ◽  
Brain Heart Infusion ◽  
Starter Cultures ◽  
Lactobacillus Helveticus ◽  
Food Sources

Among various food sources, milk proteins remain the major vector for functional peptides endowed with several biological activities. Particularly, the proteolytic activity of lactic acid bacteria during milk fermentation has been one of the most followed strategies to produce bioactive peptides. In the present study, the exploration of the activity of several starter cultures, at different fermentation times, was firstly investigated by reversed phase-high performance liquid chromatography. Among the tested strains, Lactobacillus helveticus showed a higher proteolytic activity and it was submitted to further investigations by changing the fermentation substrate (skim milk, brain heart infusion, peptone water) as well as the extraction strategy (trichloroacetic acid vs. glass beads). The chromatographic analyses and the in vitro antioxidant and antihypertensive assays highlighted considerable differences for L. helveticus hydrolysates from different substrates, while a negligible impact by the two extraction protocols emerged. Furthermore, nano-high pressure liquid chromatography coupled with a high resolution mass spectrometry analyzer allowed the preliminary discrimination of fractions from fermented skim milk, likely responsible for the found activity. The obtained results suggest the possibility of varying the fermentation parameters in order to maximize the functional effects of the bioactive peptides.

Bacterial endosymbiont inhabiting Leucaena leucocephala leaves and their antioxidant and antidiabetic potential

2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Sridevi Chigurupati ◽  
Shantini Vijayabalan ◽  
Kesavanarayanan Krishnan Selvarajan ◽  
Ahmad Alhowail ◽  
Fatema Kauser
Keyword(s):  
Free Radical ◽  
Crude Extract ◽  
Leucaena Leucocephala ◽  
Endophytic Bacteria ◽  
Radical Scavenging ◽  
16S Rrna Gene Sequencing ◽  
Free Radical Scavenging ◽  
Rrna Gene ◽  
Standard Drug

AbstractObjectivesResearch on endosymbionts is emerging globally and is considered as a potential source of bioactive phytochemicals. The present study examines the antioxidant and antidiabetic of the endophytic crude extract isolated from Leucaena leucocephala leaves.MethodsEndophytic bacteria were isolated from the leaves of L. leucocephala and 16S rRNA gene sequencing was used to establish their identity. The in vitro antioxidant effect of endophytic crude extract (LL) was evaluated using 2-diphenyl-1-picrylhydrazyl (DPPH) and 2, 2′-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) free radical scavenging methods. The in vitro antidiabetic properties of LL were evaluated using α-amylase and α-glucosidase enzyme inhibition assay.ResultsThe isolated endophytic bacteria were identified as Cronobacter sakazakii. LL displayed potent free radical scavenging effect against ABTS and DPPH radicals with an inhibitory concentration 50% (IC50) value of 17.49 ± 0.06 and 11.3 ± 0.1 μg/mL respectively. LL exhibited α-amylase and α-glucosidase inhibition with an IC50 value of 23.3 ± 0.08 and 23.4 ± 0.1 μg/mL respectively compared to the standard drug (acarbose). Both glucose loaded normoglycemic rats and STZ induced diabetic rats treated with LL (200 mg/kg) exhibited a considerable reduction in blood glucose levels p<0.01 after 8 h of treatment when compared to normal and diabetic control rats respectively.ConclusionsThus, the study shows that LL has a wellspring of natural source of antioxidants, and antidiabetic agents and phytoconstituents present in endophytes could be the rich source for bioactive compounds.

scholarly journals “Masato de Yuca” and “Chicha de Siete Semillas” Two Traditional Vegetable Fermented Beverages from Peru as Source for the Isolation of Potential Probiotic Bacteria

2021 ◽  
Author(s):  
Teresa D. Rebaza-Cardenas ◽  
Kenneth Silva-Cajaleón ◽  
Carlos Sabater ◽  
Susana Delgado ◽  
Nilda D. Montes-Villanueva ◽  
...  
Keyword(s):  
Gastrointestinal Transit ◽  
Probiotic Strain ◽  
In Vitro Digestion ◽  
Intestinal Cell ◽  
Adhesion Ability ◽  
Intestinal Cell Line ◽  
Carbohydrate Fermentation ◽  
Traditional Vegetable ◽  
Genetic Profiles

AbstractIn this work, two Peruvian beverages “Masato de Yuca,” typical of the Amazonian communities made from cassava (Manihot esculenta), and “Chicha de Siete Semillas,” made from different cereal, pseudo-cereal, and legume flours, were explored for the isolation of lactic acid bacteria after obtaining the permission of local authorities following Nagoya protocol. From an initial number of 33 isolates, 16 strains with different RAPD- and REP-PCR genetic profiles were obtained. In Chicha, all strains were Lactiplantibacillus plantarum (formerly Lactobacillus plantarum), whereas in Masato, in addition to this species, Limosilactobacillus fermentum (formerly Lactobacillus fermentum), Pediococcus acidilactici, and Weissella confusa were also identified. Correlation analysis carried out with their carbohydrate fermentation patterns and enzymatic profiles allowed a clustering of the lactobacilli separated from the other genera. Finally, the 16 strains were submitted to a static in vitro digestion (INFOGEST model) that simulated the gastrointestinal transit. Besides, their ability to adhere to the human epithelial intestinal cell line HT29 was also determined. Following both procedures, the best probiotic candidate was Lac. plantarum Ch13, a robust strain able to better face the challenging conditions of the gastrointestinal tract and showing higher adhesion ability to the intestinal epithelium in comparison with the commercial probiotic strain 299v. In order to characterize its benefit for human health, this Ch13 strain will be deeply studied in further works.

scholarly journals Root canal curvature as a prognostic factor influencing the diagnostic accuracy of radiographic working length determination and postoperative canal axis modification: an in vitro comparative study

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Bestoon Mohammed Faraj
Keyword(s):  
Root Canal ◽  
Radiographic Images ◽  
The Real ◽  
Root Canals ◽  
Length Estimation ◽  
Working Length ◽  
Length Determination ◽  
Case Difficulty Assessment ◽  
The Impact

Abstract Background Radiographic analysis of tooth morphology is mandatory for accurate calibration of the degree of canal curvature angle and radiographic working length to its real dimensions in case difficulty assessment protocols. This study aimed to determine the impact of the degree of root canal curvature angle on maintaining the real working length and the original canal axis of prepared root canals using a reciprocating rotary instrumentation technique. Methods Radiographic image analysis was performed on 60 extracted single-rooted human premolar teeth with a moderate canal curvature (10°–25°) and severe canal curvature (26°–70°). Working length and longitudinal canal axis were determined using cone-beam computed tomography (CBCT) and digital periapical radiography. The real canal length was determined by subtracting 0.5 mm from the actual canal length. Root canals were prepared using the WaveOne Gold reciprocating file (Dentsply Maillefer, Ballaigues, Switzerland). Results There was no significant relation of the degree of canal curvature angle to the accuracy of radiographic working length estimated on CBCT and digital periapical radiographic techniques (P > 0.05). Postinstrumentation changes in the original canal axis between moderate and severe canal curvature angles, assessed on CBCT and periapical digital radiographic images were statistically non-significant (P > 0.05). Conclusions A standardized digital periapical radiographic method performed similarly to the CBCT technique near to its true working length. No significant interaction exists between the diagnostic working length estimation, postoperative root canal axis modification, and the degree of canal curvature angle, using reciprocating rotary instrumentation technique.

Sign in / Sign up

Export Citation Format

Share Document