Immobilised antimicrobial peptides in downregulation of biofilm.

Colonisation of sessile bacterial species on biotic and abiotic surfaces is responsible for the development of various infections in humans. At present, biofilm-associated chronic infections have been a prime concern among the healthcare practitioners since they are impermeable to drugs, resulting in the development of antibiotic resistance or multi-drug resistance. For a few decades, a lot of research activity has been performed in the development of alternative therapeutics to combat biofilm-associated chronic infections. The presence of extracellular polymeric substance (EPS) prevents the permeation of most of the drugs rendering drug failures. The use of small molecules has been necessary to penetrate easily through the EPS and act on the targeted cells. In present days, the use of antimicrobial peptides (AMPs) has gained immense importance as alternative therapeutics since they exhibit a novel class of antibiotics exhibiting a wide spectrum of activity and possess a low rate of development of resistance. In the last few decades, a large number of AMPs have been identified from varied groups of organisms as effector molecules for innate immune system acting as an important line of defence. In this review, we will discuss the use of AMPs as effective agents to combat various biofilm-associated chronic infections.

Analysis of Antibiofilm Activities of Bioactive Compounds from Honeyweed (Leonurus sibiricus) Against P. aeruginosa: an In Vitro and In Silico Approach.

Leonurus sibiricus (Red verticilla, honeyweed) is a type of herbaceous plant predominantly found in Asian subcontinents as weed in crop fields and is widely used for treating diabetes, bronchitis, and menstrual irregularities. However, there is a dearth of study in the application of the plant phytocompounds for treating biofilm-associated chronic infections. The bioactive compounds mainly comprise of tri-terpenes, di-terpenes, phenolic acid, and flavonoids which may have potential role as antimicrobial and antibiofilm agents. Acute and chronic infection causing microbes usually form biofilm and develop virulence factors and antibiotic resistance through quorum sensing (QS). In this study, the bioactive compounds leosibirin, sibiricinone A, leosibirone A, leonotin, quercetin, lavandulifolioside, and myricetin were identified using GC-MS analysis. These were used for analyzing the antibiofilm and anti-quorum sensing activities (rhamnolipid, AHL assay, swarming motility assay) against the biofilm formed by Pseudomonas aeruginosa, the most significant nosocomial disease-causing bacteria. The compounds were able to bring about maximum inhibition in biofilm formation and QS. Although the antibiofilm activity of the phytoextract was found to be higher than that of individual phytocompounds at a concentration of 250 µg/mL, quercetin and myricetin showed highest antibiofilm activity against Pseudomonas aeruginosa, respectively, at MIC values of 135 µg/mL and 150 µg/mL against P aeruginosa. FT-IR study also revealed that the active ingredients were able to bring about the destruction of exopolysaccharides (EPS). These observations were further validated by molecular docking interactions that showed the active ingredients inhibit the functioning of QS sensing proteins by binding with them. It was observed that myricetin showed better interactions with the QS proteins of P. aeruginosa. Myricetin and quercetin show considerable inhibition of biofilm in comparison to the phytocompounds. Thus, the present study suggests that the active compounds from L. sibiricus can be used as an alternate strategy in inhibiting the biofilm formed by pathogenic organisms.

Purification, Characterization, and Application of Endoglucanase from Rhizopus oryzae as Antibiofilm Agent.

The enzyme endoglucanase is responsible for the depolymerization of cellulose. This study focuses on characterization and purification of endoglucanase from Rhizopus oryzae MTCC 9642 through a simple size exclusion method and its effective application as an antibiofilm agent. Extracellular ß-1,4-endoglucanase, an enzyme that catalyzes the hydrolysis of carboxymethyl cellulose, was found to be synthesized by Rhizopus oryzae MTCC 9642. The enzyme was purified up to homogeneity simply by size exclusion process through ultrafiltration and gel chromatography. The molecular weight of purified enzyme protein was estimated to be 39.8 kDa and it showed the highest substrate affinity towards carboxymethyl-cellulose with Km and Vmax values of 0.833 mg ml-1 and of 0.33 mmol glucose min-1 mg-1protein, respectively. The purified enzyme exhibited optimal activity at pH 6 with a broad stability range of pH 3-8. The most preferred temperature was 35 °C and 50% of activity could be retained after the thermal exposure at 40 °C for 25 min. The purified enzyme protein was inactivated by Cu2+, while the activity could be enhanced by the addition of exogenous thiols. Since biofilm is a challenge for health sector, with the aim of eradicating the biofilm, the purified endoglucanase was used to remove biofilm produced by two nosocomial bacteria. As predicted by in silico molecular docking interaction, the purified enzyme could effectively degrade biofilm architecture of bacterial strains S. aureus and P. aeruginosa by 76.52 ± 6.52% and 61.67 ± 8.76%, respectively. The properties of purified enzyme protein, as elucidated by in vitro and in silico characterization, may be favourable for its commercial applications as a potent antibiofilm agent.

Bacteriocin: A natural approach for food safety and food security.

The call to cater for the hungry is a worldwide problem in the 21st century. Food security is the utmost prime factor for the increasing demand for food. Awareness of human health when using chemical preservatives in food has increased, resulting in the use of alternative strategies for preserving food and enhancing its shelf-life. New preservatives along with novel preservation methods have been instigated, due to the intensified demand for extended shelf-life, along with prevention of food spoilage of dairy products. Bacteriocins are the group of ribosomally synthesized antimicrobial peptides; they possess a wide range of biological activities, having predominant antibacterial activity. The bacteriocins produced by the lactic acid bacteria (LAB) are considered to be of utmost importance, due to their association with the fermentation of food. In recent times among various groups of bacteriocins, leaderless and circular bacteriocins are gaining importance, due to their extensive application in industries. These groups of bacteriocins have been least studied as they possess peculiar structural and biosynthetic mechanisms. They chemically possess N-to-C terminal covalent bonds having a predominant peptide background. The stability of the bacteriocins is exhibited by the circular structure. Up till now, very few studies have been performed on the molecular mechanisms. The structural genes associated with the bacteriocins can be combined with the activity of various proteins which are association with secretion and maturation. Thus the stability of the bacteriocins can be used effectively in the preservation of food for a longer period of time. Bacteriocins are thermostable, pH-tolerant, and proteolytically active in nature, which make their usage convenient to the food industry. Several research studies are underway in the domain of biopreservation which can be implemented in food safety and food security.

Multi-Omics Approach in Amelioration of Food Products.

Determination of the quality of food products is an essential key factor needed for safe-guarding the quality of food for the interest of the consumers, along with the nutritional and sensory improvements that are necessary for delivering better quality products. Bacteriocins are a group of ribosomally synthesized antimicrobial peptides that help in maintaining the quality of food. The implementation of multi-omics approach has been important for the overall enhancement of the quality of the food. This review uses various recent technologies like proteomics, transcriptomics, and metabolomics for the overall enhancement of the quality of food products. The matrix associated with the food products requires the use of sophisticated technologies that help in the extraction of a large amount of information necessary for the amelioration of the food products. This review would provide a wholesome view of how various recent technologies can be used for improving the quality food products and for enhancing their shelf-life.

Bacterial Cellulose: Production, Characterization, and Application as Antimicrobial Agent.

Bacterial cellulose (BC) is recognized as a multifaceted, versatile biomaterial with abundant applications. Groups of microorganisms such as bacteria are accountable for BC synthesis through static or agitated fermentation processes in the presence of competent media. In comparison to static cultivation, agitated cultivation provides the maximum yield of the BC. A pure cellulose BC can positively interact with hydrophilic or hydrophobic biopolymers while being used in the biomedical domain. From the last two decades, the reinforcement of biopolymer-based biocomposites and its applicability with BC have increased in the research field. The harmony of hydrophobic biopolymers can be reduced due to the high moisture content of BC in comparison to hydrophilic biopolymers. Mechanical properties are the important parameters not only in producing green composite but also in dealing with tissue engineering, medical implants, and biofilm. The wide requisition of BC in medical as well as industrial fields has warranted the scaling up of the production of BC with added economy. This review provides a detailed overview of the production and properties of BC and several parameters affecting the production of BC and its biocomposites, elucidating their antimicrobial and antibiofilm efficacy with an insight to highlight their therapeutic potential.

Biodegradation of used polyethylene bags by a new marine strain of Alcaligenes faecalis LNDR-1.

Disposable plastic bags of two different chemical compositions and colors were remediated by the application of novel mesophilic group of bacteria isolated from the banks of sea water, using a 10 week soil burial method. The new strain, LNDR-1, was identified as Alcaligens faecalis by its morphological features and 16S rRNA sequencing. LNDR-1 was able to produce extracellular enzymes such as lipase, CMCase, xylanase, and protease, having PET surface degrading activity. It was found that LNDR-1 had a better decay rate of 15.25 ± 1% and 21.72 ± 2.1% for black and white plastic bags respectively in 10 weeks without prior oxidation as compared to S. marcescens. Polyethylene degradation was confirmed by substantial weight loss, alterations in surface topology, and hydrophobicity index and was found to be directly proportional to the ability to form biofilm on the plastic surface. FTIR results suggest presence of different metabolites in the bags treated with bacterial biofilm in comparison to the control setup inferring various types of metabolic pathways. Present study also reveals the ability of the strain to utilize the used polyethylene bag as the carbon source, without any prior treatment, and as per the literature survey, the working strain is with the capacity to biodegrade plastic at a considerably appreciable rate. This study suggests effectual method for the mechanism of biodegradation of plastic mediated by extracellular enzymes and formation of biofilm.

Antibiofilm Activity of α-Amylase from Bacillus subtilis and Prediction of the Optimized Conditions for Biofilm Removal by Response Surface Methodology (RSM) and Artificial Neural Network (ANN).

α-amylase is known to have antibiofilm activity against biofilms of both Gram positive and Gram-negative bacterial strains. Partially purified α-amylase from Bacillus subtilis was found to have inhibit biofilm formed by P. aeruginosa and S. aureus. The spectrophotometric and microscopic studies revealed that the antibiofilm efficacy of the working strain is greater than commercially purchased α-amylase. Response surface methodology (RSM) and artificial neural network (ANN) help to predict the optimum conditions [pH 8, treatment time 6 h and enzyme concentration (200 µg/mL)] for maximum biofilm eradication. This was confirmed by several in vitro experiments. Molecular docking interactions of α-amylase with the extracellular polymeric substances (EPS) of both P. aeruginosa and S. aureus indicate towards the existence of an efficient energy driven spontaneous process. Thus, this study highlights a combination of experimental and computational approach showing the naturally extracted α-amylase from B. subtilis having the potency of removing the biofilms of harmful bacterial strains involved in causing various nosocomial infections.

Catechin as the Most Efficient Bioactive Compound from Azadirachta indica with Antibiofilm and Anti-quorum Sensing Activities Against Dental Biofilm: an In Vitro and In Silico Study.

Neem (Azadirachta indica [AI]) is a unique and traditional source of antioxidant and antibacterial compounds. The GC-MS studies revealed that phytoextract of Azadirachta indica comprises a large number of phytocompounds that possess the efficacy of inhibiting the biofilm. It was observed that phytocompounds like catechin showed maximum eradication of biofilm along with the degradation of EPS structural components like carbohydrates and proteins compared to quercetin, nimbolide, nimbin, and azardirachtin, and hence, catechin was proved to be the best against dental plaque-forming bacteria. It was also observed that catechin was able to bring about a marked reduction in quorum sensing (QS) both in Alcaligenes faecalis and Pseudomonas gingivalis dental biofilm-forming strains. The extent of such reduction was maximum for catechin (94.56±2.56% in P. gingivalis & 96.56±2.5 in A. faecalis) in comparison to other bioactive compounds. It was further observed that the bioactive compounds possess the ability to quickly pass across the membrane and bring about inhibition in the DNA and RNA content of the sessile cells. This was further validated by microscopic and in silico studies. Thus, this study revealed that catechin obtained from the phytoextract of AI showed a marked ability to inhibit the dental biofilm and can be used as a natural drug-like compound in treating biofilm-associated chronic infections.