Calcium protects DNase I from proteinase K: A new method for the removal of contaminating RNase from DNase I

1980 ◽  
Vol 107 (1) ◽  
pp. 260-264 ◽  
Author(s):  
Richard H. Tullis ◽  
Harvey Rubin
Keyword(s):  
Dnase I ◽  
New Method ◽  
Proteinase K

scholarly journals Strain variation in Bacillus cereus biofilms and their susceptibility to extracellular matrix-degrading enzymes

PLoS ONE ◽  
2021 ◽  
Vol 16 (6) ◽  
pp. e0245708
Author(s):  
Eun Seob Lim ◽  
Seung-Youb Baek ◽  
Taeyoung Oh ◽  
Minseon Koo ◽  
Joo Young Lee ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Bacillus Cereus ◽  
Biofilm Formation ◽  
Dnase I ◽  
Extracellular Dna ◽  
Proteinase K ◽  
Strain Variation ◽  
Protein Ratio ◽  
Degrading Enzymes ◽  
Matrix Degrading Enzymes

Bacillus cereus is a foodborne pathogen and can form biofilms on food contact surfaces, which causes food hygiene problems. While it is necessary to understand strain-dependent variation to effectively control these biofilms, strain-to-strain variation in the structure of B. cereus biofilms is poorly understood. In this study, B. cereus strains from tatsoi (BC4, BC10, and BC72) and the ATCC 10987 reference strain were incubated at 30°C to form biofilms in the presence of the extracellular matrix-degrading enzymes DNase I, proteinase K, dispase II, cellulase, amyloglucosidase, and α-amylase to assess the susceptibility to these enzymes. The four strains exhibited four different patterns in terms of biofilm susceptibility to the enzymes as well as morphology of surface-attached biofilms or suspended cell aggregates. DNase I inhibited the biofilm formation of strains ATCC 10987 and BC4 but not of strains BC10 and BC72. This result suggests that some strains may not have extracellular DNA, or their extracellular DNA may be protected in their biofilms. In addition, the strains exhibited different patterns of susceptibility to protein- and carbohydrate-degrading enzymes. While other strains were resistant, strains ATCC 10987 and BC4 were susceptible to cellulase, suggesting that cellulose or its similar polysaccharides may exist and play an essential role in their biofilm formation. Our compositional and imaging analyses of strains ATCC 10987 and BC4 suggested that the physicochemical properties of their biofilms are distinct, as calculated by the carbohydrate to protein ratio. Taken together, our study suggests that the extracellular matrix of B. cereus biofilms may be highly diverse and provides insight into the diverse mechanisms of biofilm formation among B. cereus strains.

scholarly journals Strain variation in Bacillus cereus biofilms and their susceptibility to extracellular matrix-degrading enzymes

2021 ◽  
Author(s):  
Eun Seob Lim ◽  
Seung-Youb Baek ◽  
Taeyoung Oh ◽  
Minseon Koo ◽  
Joo Young Lee ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Bacillus Cereus ◽  
Biofilm Formation ◽  
Dnase I ◽  
Extracellular Dna ◽  
Proteinase K ◽  
Strain Variation ◽  
Protein Ratio ◽  
Degrading Enzymes ◽  
Matrix Degrading Enzymes

Bacillus cereus is a foodborne pathogen and can form biofilms on food contact surfaces, which causes food hygiene problems. While it is necessary to understand strain-dependent variation to effectively control these biofilms, strain-to-strain variation in the structure of B. cereus biofilms is poorly understood. In this study, B. cereus strains from tatsoi and the ATCC 10987 reference strain were incubated at 30? to form biofilms in the presence of the extracellular matrix-degrading enzymes DNase I, proteinase K, dispase II, cellulase, amyloglucosidase, and α-amylase to assess the susceptibility to these enzymes. The four strains exhibited four different patterns in terms of biofilm susceptibility to the enzymes as well as morphology of surface-attached biofilms or suspended cell aggregates. DNase I inhibited the biofilm formation of strains ATCC 10987 and BC4 but not of strains BC10 and BC72. This result suggests that some strains may not have extracellular DNA, or their extracellular DNA may be protected in their biofilms. In addition, the strains exhibited different patterns of susceptibility to protein- and carbohydrate-degrading enzymes. While other strains were resistant, strains ATCC 10987 and BC4 were susceptible to cellulase, suggesting that cellulose or its similar polysaccharides may exist and play an essential role in their biofilm formation. Our compositional analysis of strains ATCC 10987 and BC4 suggested that the physicochemical properties of their biofilms are distinct, as calculated by the carbohydrate to protein ratio. Taken together, our study suggests that the extracellular matrix of B. cereus biofilms may be highly diverse and provides insight into the diverse mechanisms of biofilm formation among B. cereus strains.

scholarly journals Facile Biofilm Penetration of Cationic Liposomes Loaded with DNase I/Proteinase K to Eradicate Cutibacterium acnes for Treating Cutaneous and Catheter Infections

2021 ◽  
Vol Volume 16 ◽  
pp. 8121-8138
Author(s):  
Jia-You Fang ◽  
Wei-Ling Chou ◽  
Chwan-Fwu Lin ◽  
Calvin T Sung ◽  
Ahmed Alalaiwe ◽  
...  
Keyword(s):  
Cationic Liposomes ◽  
Dnase I ◽  
Proteinase K ◽  
Cutibacterium Acnes ◽  
Catheter Infections

scholarly journals New Method of Plague Agent Lipopolysaccharide Obtaining

2014 ◽  
pp. 100-103
Author(s):  
T. A. Polunina ◽  
N. P. Guseva ◽  
I. A. Kuz’Michenko ◽  
Z. L. Devdariani ◽  
S. P. Zadnova ◽  
...  
Keyword(s):  
Salt Water ◽  
Chemical Properties ◽  
New Method ◽  
Proteinase K ◽  
Vaccine Production ◽  
Commercial Preparation ◽  
Phenol Extraction ◽  
Cost Cutting ◽  
Physical Chemical ◽  
Triton X

Put forward are two alternatives of a new method for optimization of conditions of LPS obtaining and purification from Y. pestis strains; as well as for avoiding application of poisonous and hard-to-remove reagents; for simplification and cost-cutting of the technique; and for rationalization of production waste management. This method involves preliminary salt-water extraction of bacteria, for elimination of easy-dissolving substances, with the subsequent fracturing using ultrasound in lysing buffer (0,1 M Tris-HCl, pH 8,0; 10 mmol of EDTA, 1 % Triton X-100). The first alternative for deproteinization of non-purified endotoxin is the commercial preparation of proteinase K (Sigma), the second one - an enzyme complex - proteovibrin, isolated from waste material accumulated in the process of cholera chemical bivalent vaccine production. Apart from this, introduced has been a phase of sample acidification by applying glacial acetic acid up to pH 3,2-3,4 to decontaminate LPS from nucleic acids. These two variations of the method provide for enhancement of LPS preparation quality as compared to prototype method, and for obtainment of plague agent endotoxin that is hardly distinguishable in physical-chemical properties, homogeneity, immunochemical activity and specificity from the antigen, manufactured by means of water-phenol extraction following Westphal O. technique.

scholarly journals Single DNase or Proteinase Treatment Induces Change in Composition and Structural Integrity of Multispecies Oral Biofilms

Antibiotics ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 400
Author(s):  
Lamprini Karygianni ◽  
Pune N. Paqué ◽  
Thomas Attin ◽  
Thomas Thurnheer
Keyword(s):  
Laser Scanning ◽  
Structural Integrity ◽  
Dnase I ◽  
Laser Scanning Microscopy ◽  
Proteinase K ◽  
Confocal Laser ◽  
Significance Level ◽  
Biofilm Thickness ◽  
Biofilm Control ◽  
Oral Biofilms

Biofilm virulence is mainly based on its bacterial cell surrounding biofilm matrix, which contains a scaffold of exopolysaccharides, carbohydrates, proteins, lipids, and nucleic acids. Targeting these nucleid acids or proteins could enable an efficient biofilm control. Therefore, the study aimed to test the effect of deoxyribonuclease I (DNase I) and proteinase K on oral biofilms. Six-species biofilms (Streptococcus mutans, Streptococcus oralis, Actinomyces oris, Fusobacterium nucleatum, Veillonella dispar, and Candida albicans) were exposed to DNase I (0.001 mg/mL, 0.002 mg/mL) or proteinase K (0.05 mg/mL, 0.1 mg/mL) for 1 h during biofilm formation. After 64 h, biofilms were harvested, quantified by culture analysis and visualized by image analysis using CLSM (confocal laser scanning microscopy). Statistical analysis was performed by ANOVA, followed by the Tukey test at a 5% significance level. The biofilm treatment with proteinase K induced a significant increase of Logs10 counts in S. mutans and a decrease in C. albicans, while biofilm thickness was reduced from 28.5 μm (control) to 9.07 μm (0.05 mg/mL) and 7.4 μm (0.1 mg/mL). Treatment with DNase I had no effect on the total bacterial growth within the biofilm. Targeting proteins of biofilms by proteinase K are promising adjunctive tool for biofilm control.

scholarly journals Combined DNase and Proteinase Treatment Interferes with Composition and Structural Integrity of Multispecies Oral Biofilms

2020 ◽  
Vol 9 (4) ◽  
pp. 983 ◽  
Author(s):  
Lamprini Karygianni ◽  
Thomas Attin ◽  
Thomas Thurnheer
Keyword(s):  
Laser Scanning ◽  
Antimicrobial Agents ◽  
Structural Integrity ◽  
Dnase I ◽  
Laser Scanning Microscopy ◽  
Proteinase K ◽  
Confocal Laser ◽  
Dental Hard Tissues ◽  
Oral Biofilms ◽  
The Impact

Modification of oral biofilms adhering to dental hard tissues could lead to new treatment approaches in cariology and periodontology. In this study the impact of DNase I and/or proteinase K on the formation of a simulated supragingival biofilm was investigated in vitro. Six-species biofilms were grown anaerobically in the presence of DNase I and proteinase K. After 64 h biofilms were either harvested and quantified by culture analysis or proceeded to staining followed by confocal laser scanning microscopy. Microbial cells were stained using DNA-dyes or fluorescent in situ hybridization. Exopolysaccharides, eDNA and exoproteins were stained with Calcofluor, anti-DNA-antibody, and SyproTM Ruby, respectively. Overall, results showed that neither DNase I nor proteinase K had an impact on total colony-forming units (CFUs) compared to the control without enzymes. However, DNase I significantly suppressed the growth of Actinomyces oris, Fusobacterium nucleatum, Streptococcus mutans, Streptococcus oralis and Candida albicans. Proteinase K treatment induced significant increase in S. mutans and S. oralis CFUs (p < 0.001), whereas C. albicans and V. dispar showed lower CFUs compared to the control. Interestingly, confocal images visualized the biofilm degradation caused by DNase I and proteinase K. Thus, enzymatic treatment should be combined with conventional antimicrobial agents aiming at both bactericidal effectiveness and biofilm dispersal.

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