Efficient Biofilms Eradication by Enzymatic-Cocktail of Pancreatic Protease Type-I and Bacterial α-Amylase

Seung-Cheol Jee; Min Kim; Jung-Suk Sung; Avinash A. Kadam

doi:10.3390/polym12123032

Efficient Biofilms Eradication by Enzymatic-Cocktail of Pancreatic Protease Type-I and Bacterial α-Amylase

Polymers ◽

10.3390/polym12123032 ◽

2020 ◽

Vol 12 (12) ◽

pp. 3032

Author(s):

Seung-Cheol Jee ◽

Min Kim ◽

Jung-Suk Sung ◽

Avinash A. Kadam

Keyword(s):

Type I ◽

Enzymatic System ◽

Biofilm Inhibition ◽

E Coli ◽

Inhibition Effect ◽

Enzyme Combination ◽

Pancreatic Protease ◽

Bovine Pancreas ◽

First Time ◽

Biofilm Prevention

Removal of biofilms is extremely pivotal in environmental and medicinal fields. Therefore, reporting the new-enzymes and their combinations for dispersal of infectious biofilms can be extremely critical. Herein, for the first time, we accessed the enzyme “protease from bovine pancreas type-I (PtI)” for anti-biofilm properties. We further investigated the anti-biofilm potential of PtI in combination with α-amylase from Bacillus sp. (αA). PtI showed a very significant biofilm inhibition effect (86.5%, 88.4%, and 67%) and biofilm prevention effect (66%, 64%, and 70%), against the E. coli, S. aureus, and MRSA, respectively. However, the new enzyme combination (Ec-PtI+αA) exhibited biofilm inhibition effect (78%, 90%, and 93%) and a biofilm prevention effect (44%, 51%, and 77%) against E. coli, S. aureus, and MRSA, respectively. The studied enzymes were found not to be anti-bacterial against the E. coli, S. aureus, and MRSA. In summary, the PtI exhibited significant anti-biofilm effects against S. aureus, MRSA, and E. coli. Ec-PtI+αA exhibited enhancement of the anti-biofilm effects against S. aureus and MRSA biofilms. Therefore, this study revealed that this Ec-PtI+αA enzymatic system can be extremely vital for the treatment of biofilm complications resulting from E. coli, S. aureus, and MRSA.

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Grappling with anisotropic data, pseudo-merohedral twinning and pseudo-translational noncrystallographic symmetry: a case study involving pyruvate kinase

Acta Crystallographica Section D Structural Biology ◽

10.1107/s205979831600142x ◽

2016 ◽

Vol 72 (4) ◽

pp. 512-519 ◽

Cited By ~ 7

Author(s):

Katherine A. Donovan ◽

Sarah C. Atkinson ◽

Sarah A. Kessans ◽

Fen Peng ◽

Tim F. Cooper ◽

...

Keyword(s):

Pyruvate Kinase ◽

Orthorhombic Symmetry ◽

Type I ◽

Space Group ◽

E Coli ◽

Crystallization Conditions ◽

Lid Domain ◽

Fructose 1,6 Bisphosphate ◽

First Time

Pyruvate kinase is a key regulatory enzyme involved in the glycolytic pathway. The crystal structure ofEscherichia colitype I pyruvate kinase was first solved in 1995 at 2.5 Å resolution. However, the space group was ambiguous, being either primitive orthorhombic (P212121) orC-centred orthorhombic (C2221). Here, the structure determination and refinement ofE. colitype I pyruvate kinase to 2.28 Å resolution are presented. Using the same crystallization conditions as reported previously, the enzyme was found to crystallize in space groupP21. Determination of the space group was complicated owing to anisotropic data, pseudo-translational noncrystallographic symmetry and the pseudo-merohedrally twinned nature of the crystal, which was found to have very close to 50% twinning, leading to apparent orthorhombic symmetry and absences that were not inconsistent withP212121. The unit cell contained two tetramers in the asymmetric unit (3720 residues) and, when compared with the orthorhombic structure, virtually all of the residues could be easily modelled into the density. Averaging of reflections into the lower symmetry space group with twinning provided tidier electron density that allowed ∼30 missing residues of the lid domain to be modelled for the first time. Moreover, residues in a flexible loop could be modelled and sulfate molecules are found in the allosteric binding domain, identifying the pocket that binds the allosteric activator fructose 1,6-bisphosphate in this isozyme for the first time. Lastly, we note the pedagogical benefits of difficult structures to emerging crystallographers.

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An unusual type I ribosome-inactivating protein from Agrostemma githago L.

Scientific Reports ◽

10.1038/s41598-020-72282-2 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Christoph Weise ◽

Achim Schrot ◽

Leonie T. D. Wuerger ◽

Jacob Adolf ◽

Roger Gilabert-Oriol ◽

...

Keyword(s):

Current Knowledge ◽

Ricinus Communis ◽

Herbaceous Plant ◽

Type I ◽

Ribosome Inactivating Protein ◽

Ribosome Inactivating Proteins ◽

E Coli ◽

The Past ◽

First Time ◽

Unusual Type

Abstract Agrostemma githago L. (corn cockle) is an herbaceous plant mainly growing in Europe. The seeds of the corn cockle are toxic and poisonings were widespread in the past by consuming contaminated flour. The toxic principle of Agrostemma seeds was attributed to triterpenoid secondary metabolites. Indeed, this is in part true. However Agrostemma githago L. is also a producer of ribosome-inactivating proteins (RIPs). RIPs are N-glycosylases that inactivate the ribosomal RNA, a process leading to an irreversible inhibition of protein synthesis and subsequent cell death. A widely known RIP is ricin from Ricinus communis L., which was used as a bioweapon in the past. In this study we isolated agrostin, a 27 kDa RIP from the seeds of Agrostemma githago L., and determined its full sequence. The toxicity of native agrostin was investigated by impedance-based live cell imaging. By RNAseq we identified 7 additional RIPs (agrostins) in the transcriptome of the corn cockle. Agrostin was recombinantly expressed in E. coli and characterized by MALDI-TOF–MS and adenine releasing assay. This study provides for the first time a comprehensive analysis of ribosome-inactivating proteins in the corn cockle and complements the current knowledge about the toxic principles of the plant.

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Efficacy of compressed sodium chloride (CSC) against E. coli and Candida auris in minutes and methods improvement for testing

Scientific Reports ◽

10.1038/s41598-020-79212-2 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Lan N. Truong ◽

Brayden D. Whitlock

Keyword(s):

Sodium Chloride ◽

Improved Method ◽

Candida Auris ◽

Antimicrobial Surfaces ◽

E Coli ◽

Route Of Transmission ◽

The World ◽

Short Time ◽

First Time ◽

Time Point

AbstractControlling infections has become one of the biggest problems in the world, whether measured in lives lost or money spent. This is worsening as pathogens continue becoming resistant to therapeutics. Antimicrobial surfaces are one strategy being investigated in an attempt to decrease the spread of infections through the most common route of transmission: surfaces, including hands. Regulators have chosen two hours as the time point at which efficacy should be measured. The objectives of this study were to characterize the new antimicrobial surface compressed sodium chloride (CSC) so that its action may be understood at timepoints more relevant to real-time infection control, under two minutes; to develop a sensitive method to test efficacy at short time points; and to investigate antifungal properties for the first time. E. coli and Candida auris are added to surfaces, and the surfaces are monitored by contact plate, or by washing into collection vats. An improved method of testing antimicrobial efficacy is reported. Antimicrobial CSC achieves at least 99.9% reduction of E. coli in the first two minutes of contact, and at least 99% reduction of C. auris in one minute.

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Antimicrobial and Immunomodulating Activities of Two Endemic Nepeta Species and Their Major Iridoids Isolated from Natural Sources

Pharmaceuticals ◽

10.3390/ph14050414 ◽

2021 ◽

Vol 14 (5) ◽

pp. 414

Author(s):

Neda Aničić ◽

Uroš Gašić ◽

Feng Lu ◽

Ana Ćirić ◽

Marija Ivanov ◽

...

Keyword(s):

Biofilm Formation ◽

Food Industry ◽

Balkan Peninsula ◽

Antimicrobial Activities ◽

Natural Sources ◽

E Coli ◽

Food Borne ◽

Metabolite Profiles ◽

Aspergillus Sp ◽

First Time

Two Balkan Peninsula endemics, Nepeta rtanjensis and N. argolica subsp. argolica, both characterized by specialized metabolite profiles predominated by iridoids and phenolics, are differentiated according to the stereochemistry of major iridoid aglycone nepetalactone (NL). For the first time, the present study provides a comparative analysis of antimicrobial and immunomodulating activities of the two Nepeta species and their major iridoids isolated from natural sources—cis,trans-NL, trans,cis-NL, and 1,5,9-epideoxyloganic acid (1,5,9-eDLA), as well as of phenolic acid rosmarinic acid (RA). Methanol extracts and pure iridoids displayed excellent antimicrobial activity against eight strains of bacteria and seven strains of fungi. They were especially potent against food-borne pathogens such as L. monocytogenes, E. coli, S. aureus, Penicillium sp., and Aspergillus sp. Targeted iridoids were efficient agents in preventing biofilm formation of resistant P. aeruginosa strain, and they displayed additive antimicrobial interaction. Iridoids are, to a great extent, responsible for the prominent antimicrobial activities of the two Nepeta species, although are probably minor contributors to the moderate immunomodulatory effects. The analyzed iridoids and RA, individually or in mixtures, have the potential to be used in the pharmaceutical industry as potent antimicrobials, and in the food industry to increase the shelf life and safety of food products.

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Poly(Vinylamine) Derived N-Doped C-Dots with Antimicrobial and Antibiofilm Activities

C – Journal of Carbon Research ◽

10.3390/c7020040 ◽

2021 ◽

Vol 7 (2) ◽

pp. 40

Author(s):

Semiha Duygu Sutekin ◽

Mehtap Sahiner ◽

Selin Sagbas Suner ◽

Sahin Demirci ◽

Olgun Güven ◽

...

Keyword(s):

Blood Clotting ◽

Minimum Bactericidal Concentration ◽

Blood Compatibility ◽

Weight Ratio ◽

Gram Negative Bacteria ◽

Biofilm Inhibition ◽

E Coli ◽

Gram Positive Bacteria ◽

Doped Carbon Dots ◽

Nitrogen Doped Carbon

Nitrogen-doped carbon dots (N-doped C-dots) was synthesized by using poly(vinyl amine) (PVAm) as a nitrogen source and citric acid (CA) as a carbon source via the hydrothermal method. Various weight ratios of CA and PVAm (CA:PVAm) were used to synthesize N-doped C-dots. The N-doped C-dots revealed emission at 440 nm with excitation at 360 nm and were found to increase the fluorescence intensity with an increase in the amount of PVAm. The blood compatibility studies revealed no significant hemolysis for N-doped C-dots that were prepared at different ratios of CA:PVAm for up to 500 μg/mL concentration with the hemolysis ratio of 1.96% and the minimum blood clotting index of 88.9%. N-doped C-dots were found to be more effective against Gram-positive bacteria than Gram-negative bacteria, with the highest potency on Bacillus subtilis (B. subtilis). The increase in the weight ratio of PVAm in feed during C-dots preparation from 1 to 3 leads to a decrease of the minimum bactericidal concentration (MBC) value from 6.25 to 0.75 mg/mL for B. subtilis. Antibiofilm ability of N-doped C-dots prepared by 1:3 ratio of CA:PVAm was found to reduce %biofilm inhibition and eradication- by more than half, at 0.78 mg/mL for E. coli and B. subtilis generated biofilms and almost destroyed at 25 mg/mL concentrations.

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Radical Dehalogenation and Purine Nucleoside Phosphorylase E. coli: How Does an Admixture of 2′,3′-Anhydroinosine Hinder 2-fluoro-cordycepin Synthesis

Biomolecules ◽

10.3390/biom11040539 ◽

2021 ◽

Vol 11 (4) ◽

pp. 539

Author(s):

Alexey L. Kayushin ◽

Julia A. Tokunova ◽

Ilja V. Fateev ◽

Alexandra O. Arnautova ◽

Maria Ya. Berzina ◽

...

Keyword(s):

Nmr Spectroscopy ◽

Chemical Synthesis ◽

Purine Nucleoside Phosphorylase ◽

Purine Nucleoside ◽

Preparative Synthesis ◽

Nucleoside Phosphorylase ◽

E Coli ◽

First Time

During the preparative synthesis of 2-fluorocordycepin from 2-fluoroadenosine and 3′-deoxyinosine catalyzed by E. coli purine nucleoside phosphorylase, a slowdown of the reaction and decrease of yield down to 5% were encountered. An unknown nucleoside was found in the reaction mixture and its structure was established. This nucleoside is formed from the admixture of 2′,3′-anhydroinosine, a byproduct in the preparation of 3-′deoxyinosine. Moreover, 2′,3′-anhydroinosine forms during radical dehalogenation of 9-(2′,5′-di-O-acetyl-3′-bromo- -3′-deoxyxylofuranosyl)hypoxanthine, a precursor of 3′-deoxyinosine in chemical synthesis. The products of 2′,3′-anhydroinosine hydrolysis inhibit the formation of 1-phospho-3-deoxyribose during the synthesis of 2-fluorocordycepin. The progress of 2′,3′-anhydroinosine hydrolysis was investigated. The reactions were performed in D2O instead of H2O; this allowed accumulating intermediate substances in sufficient quantities. Two intermediates were isolated and their structures were confirmed by mass and NMR spectroscopy. A mechanism of 2′,3′-anhydroinosine hydrolysis in D2O is fully determined for the first time.

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Modular invariant models of leptons at level 7

Journal of High Energy Physics ◽

10.1007/jhep08(2020)164 ◽

2020 ◽

Vol 2020 (8) ◽

Cited By ~ 4

Author(s):

Gui-Jun Ding ◽

Stephen F. King ◽

Cai-Chang Li ◽

Ye-Ling Zhou

Keyword(s):

Modular Forms ◽

Galois Field ◽

Special Linear Group ◽

Type I ◽

Time Level ◽

Projective Special Linear Group ◽

Modular Invariant ◽

Seesaw Mechanism ◽

Linearly Independent ◽

First Time

Abstract We consider for the first time level 7 modular invariant flavour models where the lepton mixing originates from the breaking of modular symmetry and couplings responsible for lepton masses are modular forms. The latter are decomposed into irreducible multiplets of the finite modular group Γ7, which is isomorphic to PSL(2, Z7), the projective special linear group of two dimensional matrices over the finite Galois field of seven elements, containing 168 elements, sometimes written as PSL2(7) or Σ(168). At weight 2, there are 26 linearly independent modular forms, organised into a triplet, a septet and two octets of Γ7. A full list of modular forms up to weight 8 are provided. Assuming the absence of flavons, the simplest modular-invariant models based on Γ7 are constructed, in which neutrinos gain masses via either the Weinberg operator or the type-I seesaw mechanism, and their predictions compared to experiment.

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Antimicrobial Activity of L-Lysine and Poly-L-Lysine with Pulsed Electric Fields

Applied Sciences ◽

10.3390/app11062708 ◽

2021 ◽

Vol 11 (6) ◽

pp. 2708

Author(s):

Jurgita Švedienė ◽

Vitalij Novickij ◽

Rokas Žalnėravičius ◽

Vita Raudonienė ◽

Svetlana Markovskaja ◽

...

Keyword(s):

Electric Fields ◽

Pulsed Electric Fields ◽

Antimicrobial Treatment ◽

Treatment Development ◽

E Coli ◽

Yeast Fungi ◽

New Treatment ◽

First Time ◽

Resistant Microorganism ◽

Susceptibility Patterns

For the first time, the possibility to use L-lysine (Lys) and poly-L-lysine (PLL) as additives with pulsed electric fields (PEF) for antimicrobial treatment is reported. The antimicrobial efficacy of Lys and PLL for Escherichia coli, Staphylococcus aureus, Trichophyton rubrum and Candida albicans was determined. Inactivation of microorganisms was also studied by combining Lys and PLL with PEF of 15 and 30 kV/cm. For PEF treatment, pulses of 0.5, 1, 10 or 100 μs were applied in a sequence of 10 to 5000 at 1 kHz frequency. The obtained results showed that 100 μs pulses were the most effective in combination with Lys and PLL for all microorganisms. Equivalent energy PEF bursts with a shorter duration of the pulse were less effective independently on PEF amplitude. Additionally, various treatment susceptibility patterns of microorganisms were determined and reported. In this study, the Gram-negative E. coli was the most treatment-resistant microorganism. Nevertheless, inactivation rates exceeding 2 log viability reduction were achieved for all analyzed yeast, fungi, and bacteria. This methodology could be used for drug-resistant microorganism’s new treatment development.

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Bioactive constituents with antibacterial, resistance modulation, anti-biofilm formation and efflux pump inhibition properties from Aidia genipiflora stem bark

Clinical Phytoscience ◽

10.1186/s40816-021-00266-4 ◽

2021 ◽

Vol 7 (1) ◽

Author(s):

Daniel Anokwah ◽

Evelyn Asante-Kwatia ◽

Abraham Y. Mensah ◽

Cynthia Amaning Danquah ◽

Benjamin K. Harley ◽

...

Keyword(s):

Biofilm Formation ◽

Efflux Pump ◽

Ethyl Acetate Fraction ◽

Stem Bark ◽

Resistance Mechanisms ◽

Bioactive Constituents ◽

Biofilm Inhibition ◽

E Coli ◽

Efflux Pump Inhibition ◽

Oleanonic Acid

Abstract Background Antimicrobial resistance is a global health challenge. The involvement of bacterial biofilms and efflux pumps in the development of multidrug resistance (MDR) is well established. Medicinal plants have been proposed as alternatives for combating MDR focusing on their bioactive constituents with resistance modulatory activities. This study was aimed at investigating the stem bark of Aidia genipiflora for bioactive constituents with anti-biofilm, efflux pump inhibition and resistance modulatory activities. Method The crude methanol extract was purified by column chromatography and isolated compounds characterized by mass and nuclear magnetic resonance spectrometry. Antibacterial activity was determined by the High-throughput spot culture growth inhibition and the broth micro-dilution assay. The ethidium bromide accumulation assay was used to determine efflux pump inhibition property. Biofilm inhibition was determined in a microplate crystal violet retention assay. Results Purification of the ethyl acetate fraction led to the isolation of oleanonic acid (1), 4-hydroxy cinnamic acid docosyl ester (2), β-stigmasterol/β-sitosterol (mixture 3a/b) and D-mannitol (4). The minimum inhibitory concentrations (MICs) ranged from 250 to > 500 μg/mL for extracts and fractions and from 15 to 250 μg/mL for compounds. In the presence of sub-inhibitory concentrations of the compounds, the MIC of amoxicillin against E. coli (20 μg/mL) and P. aeruginosa (320 μg/mL) was reduced by 32 and 10 folds respectively. The whole extract demonstrated anti-biofilm formation and efflux pump inhibition in E. coli, S. aureus and P. aeruginosa. The sterol mixture (3a/b) at concentration of 100 μg/mL caused the highest inhibition (73%) of biofilm formation in S. aureus. Oleanonic acid (1) demonstrated remarkable efflux pump inhibition at MIC of 7.8 μg/mL in E. coli better than the standard drugs verapamil and chlorpromazine. Conclusion This study confirms the prospects of A. genipiflora as a source of new antibacterial agents and adjuvants that could interact with some resistance mechanisms in bacteria to enhance the activity of hitherto ineffective antibiotics. “A small portion of the study has been presented in a conference in the form of poster”.

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