Microbial lipopeptides: their pharmaceutical and biotechnological potential, applications, and way forward.

As lead molecules, cyclic lipopeptides with antibacterial, antifungal, and antiviral properties have garnered a lot of attention in recent years. Because of their potential, cyclic lipopeptides have earned recognition as a significant class of antimicrobial compounds with applications in pharmacology and biotechnology. These lipopeptides, often with biosurfactant properties, are amphiphilic, consisting of a hydrophilic moiety, like a carboxyl group, peptide backbone, or carbohydrates, and a hydrophobic moiety, mostly a fatty acid. Besides, several lipopeptides also have cationic groups that play an important role in biological activities. Antimicrobial lipopeptides can be considered as possible substitutes for antibiotics that are conventional to address the current drug-resistant issues as pharmaceutical industries modify the parent antibiotic molecules to render them more effective against antibiotic-resistant bacteria and fungi, leading to the development of more resistant microbial strains. Bacillus species produce lipopeptides, which are secondary metabolites that are amphiphilic and are typically synthesized by non-ribosomal peptide synthetases (NRPSs). They have been identified as potential biocontrol agents as they exhibit a broad spectrum of antimicrobial activity. A further benefit of lipopeptides is that they can be produced and purified biotechnologically or biochemically in a sustainable manner using readily available, affordable, renewable sources without harming the environment. In this review, we discuss the biochemical and functional characterization of antifungal lipopeptides, as well as their various modes of action, method of production and purification (in brief), and potential applications as novel antibiotic agents.

Production and characterization of a novel poly amino acid from a thermophilic bacterium, and preliminary testing of its coagulating potential for imminent wastewater treatment application.

The increasing demand for biopolymers across diverse fields, such as food, medicine, cosmetics, and environmental applications, has prompted researchers to explore novel molecules with enhanced functionalities that meet these demands. In this study, a thermophilic strain of Bacillus licheniformis was employed to produce a unique polyamino acid. This thermophilic isolate exhibited rapid growth at 50 °C in a sucrose mineral salts medium, resulting in a biopolymer concentration of 7.4 g/L. Interestingly, the biopolymer produced at different temperatures exhibited varying glass-transition temperatures (ranging from 87.86 °C to 104.11 °C) and viscosities (7.5 cP to 16.3 cP), suggesting that the fermentation temperature significantly influenced the degree of polymerization. Furthermore, the biopolymer was characterized using various techniques, including Thin Layer Chromatography (TLC), Fourier Transform Infrared (FTIR) spectroscopy, Liquid Chromatography-Electrospray Ionization-Mass Spectroscopy (LC-ESI MS), Nuclear Magnetic Resonance (NMR), and Differential Scanning Calorimetry-Thermogravimetric Analysis (DSC-TGA). The results revealed that the obtained biopolymer was a poly amino acid, with poly-γ-glutamic acid as the major monomeric component in the polymer backbone with a few appendages of aspartic acid residues in its side chain. Finally, the biopolymer demonstrated significant coagulation potential for water treatment applications, as evidenced by coagulation studies conducted under varying pH conditions using kaolin-clay as a model precipitant.

Antimicrobial Nanoparticles Mediated Prevention and Control of Membrane Biofouling in Water and Wastewater Treatment: Current Trends and Future Perspectives.

Global water scarcity and water pollution necessitate wastewater reclamation for further use. As an alternative to conventional techniques, membrane technology is extensively used as an advanced method for water purification and wastewater treatment due to its selectivity, permeability, and efficient removal of pollutants. However, microbial biofouling is a major threat that deteriorates membrane performance and imparts operational challenges. It is a natural phenomenon caused by the undesirable colonization of microbes on membrane surfaces. The economic penalties associated with this menace are enormous. The traditional preventive measures are dominated by biocides, toxic chemicals, cleaners and antifouling surfaces, which are costly and often cause secondary pollution. Recent focus is thus being directed to promote inputs from nanotechnology to control and mitigate this major threat. Different anti-microbial nanomaterials can be effectively used to prevent the adhesion of microbes onto the membrane surfaces and eliminate microbial biofilms, to provide an economical and eco-friendly solution to biofouling. This review addresses the formation of microbial biofilms and biofouling in membrane operations. The potential of nanocomposite membranes in alleviating this problem and the challenges in commercialization are discussed. The antifouling mechanisms are also highlighted, which are not widely elucidated.

On-site enriched production of cellulase enzyme using rice straw waste and its hydrolytic performance evaluation through systematic dynamic modeling.

The application of on-site produced cellulolytic enzymes in place of commercial enzymes towards hydrolytic preparations of reducing sugars using inexpensive lignocellulosic wastes is considered the most efficient strategy to accomplish a cost-effective biofuel production process. Along with improved production, intrinsic and systematic performance evaluation of the produced enzyme during the hydrolysis process through kinetic intervention remains a crucial requirement for achieving the improved performance of the process. With this motivation, the present study primarily deals with the nutritionally optimized production strategy of cellulases from rice straw (RS) waste using Trichoderma reesei (MTCC 164). The highest cellulase production was obtained 8.09 ± 0.32 g/l in batch mode at optimized combinations of 3.5% (w/v) RS inducer, 3.0% (w/v) lactose, and 1.5% (w/v) peptone. Production was further improved through pH-regulated (pH 5.5 to 6.5) fed-batch fermentations. The enzyme produced at pH 6 was considered for hydrolysis studies at 4 to 10% (w/w) solid loading due to reasonable exoglucanase, endoglucanase, and maximum ß-glucosidase activity levels of 9.3 U/ml, 3.87 U/ml, and 2.65 U/ml respectively. Multi-reaction systematic kinetic modeling was implemented to evaluate enzyme performance during hydrolysis, and the values of inhibitory kinetic parameters (K2r = 7.1 < K1r = 18.5 < K3r = 276.6) suggested that sequential conversion of cellulose to glucose by existing enzyme components was more dominant over direct conversion.

Kinetic model supported improved and optimized submerged production strategy of cellulase enzyme from newspaper waste biomass.

A systematic evaluation of microorganism's potential towards biosynthesis of cellulases from inexpensive lignocellulosic feedstock through appropriate kinetic modelling facilitates understanding, optimization and designing of an effective industrial cellulase enzyme production process. The present study aims to optimize a submerged fungal cultivation strategy for cellulase production from abundantly available newspaper wastes (NPW). A combined pretreatment strategy consisting diluted, 1% (v v-1) H2SO4 followed by 2% (w v-1) NaOH treatment was highly effective to convert newspaper waste to an effective cellulose-enriched inducer for the production of cellulase. In addition, the composition of the most influential nutrient components like peptone and lactose was optimized with the help of response surface methodology for enhanced cellulase production with maximum activity levels. Maximum cellulase production of 8.64 g L-1 with 7.82 FPU mL-1 total activity levels was achieved from optimized composition of pretreated NPW 3.29% (w v-1), lactose 2.94% (w v-1) and peptone 1.53% (w v-1). To analyse intrinsic inhibition effect of the substrate concentration on cellulase production, modified Luedeking-Piret model simulated experiments were further conducted with 1.5% (w/v), 3.29% (w/v) and 4% (w/v) NPW concentrations. The developed kinetic model perfectly captured the trends of biomass production, substrate consumption and adsorption characteristic of cellulase enzyme on its activity during production. The rate constant for cellulase synthesis was evaluated to be increased to 0.040 IU g-1 h -1 at 3.29% (w v-1) of NPW concentration; however, it was further reduced to 0.024 IU g-1 h -1 at higher NPW concentration of 4% (w v-1).

Improved fed-batch production of high-purity PHB (poly-3 hydroxy butyrate) by Cupriavidus necator (MTCC 1472) from sucrose-based cheap substrates under response surface-optimized conditions.

Experimental investigations were carried out for Cupriavidus necator (MTCC 1472)-based improved production of poly-3 hydroxy butyrate (PHB) through induced nitrogen limiting fed-batch cultivation strategies. Initially Plackett-Burman design and response surface methodology were implemented to optimize most influencing process parameters. With optimized process parameter values, continuous feeding strategies ware applied in a 5-l fermenter with table sugar concentration of 100 g/l, nitrogen concentration of 0.12 g/l for fed-batch fermentation with varying dilution rates of 0.02 and 0.046 1/h. To get enriched production of PHB, concentration of the sugar was further increased to 150 and 200 g/l in feeding. Maximum concentrations of PHB achieved were 22.35 and 23.07 g/l at those dilution rates when sugar concentration maintains at 200 g/l in feeding. At maximum concentration of PHB (23.07 g/l), productivity of 0.58 g/l h was achieved with maximum PHB accumulation efficiency up to 64% of the dry weight of biomass. High purity of PHB, close to medical grade was achieved after surfactant hypochlorite extraction method, and it was further confirmed by SEM, EDX, and XRD studies.

Biosurfactant-biopolymer driven microbial enhanced oil recovery (MEOR) and its optimization by an ANN-GA hybrid technique.

A lipopeptide biosurfactant produced by marine Bacillus megaterium and a biopolymer produced by thermophilic Bacillus licheniformis were tested for their application potential in the enhanced oil recovery. The crude biosurfactant obtained after acid precipitation effectively reduced the surface tension of deionized water from 70.5 to 28.25mN/m and the interfacial tension between lube oil and water from 18.6 to 1.5mN/m at a concentration of 250mgL-1. The biosurfactant exhibited a maximum emulsification activity (E24) of 81.66% against lube oil. The lipopeptide micelles were stabilized by addition of Ca2+ ions to the biosurfactant solution. The oil recovery efficiency of Ca2+ conditioned lipopeptide solution from a sand-packed column was optimized by using artificial neural network (ANN) modelling coupled with genetic algorithm (GA) optimization. Three important parameters namely lipopeptide concentration, Ca2+ concentration and solution pH were considered for optimization studies. In order to further improve the recovery efficiency, a water soluble biopolymer produced by Bacillus licheniformis was used as a flooding agent after biosurfactant incubation. Upon ANN-GA optimization, 45% tertiary oil recovery was achieved, when biopolymer at a concentration of 3gL-1 was used as a flooding agent. Oil recovery was only 29% at optimal conditions predicted by ANN-GA, when only water was used as flooding solution. The important characteristics of biopolymers such as its viscosity, pore plugging capabilities and bio-cementing ability have also been tested. Thus, as a result of biosurfactant incubation and biopolymer flooding under the optimal process conditions, a maximum oil recovery of 45% was achieved. Therefore, this study is novel, timely and interesting for it showed the combined influence of biosurfactant and biopolymer on solubilisation and mobilization of oil from the soil.

Investigation on sodium benzoate release from poly(butylene adipate-co-terephthalate)/organoclay/sodium benzoate based nanocomposite film and their antimicrobial activity.

Polymeric nanocomposites embedded with nontoxic antimicrobial agents have recently gained potential industrial significance, mainly for their applicability to preserve food quality and ensure safety. In this study, a poly(butylene adipate-co-terephthalate) (PBAT)/organoclay (CMMT) based nanocomposite film doped with sodium benzoate (SB) as antimicrobial agent was prepared by a solution mixing process. A homogenous dispersion of organoclay (cetyltrimethylammonium-modified montmorillonite [CMMT]) in PBAT matrix was observed by X-ray diffraction and transmission electron microscopy. PBAT/CMMT nanocomposite film showed higher barrier properties against water and methanol vapor compared to the PBAT film. The release of SB from PBAT and its nanocomposite film was measured and the relevant data were fitted to the Weibull model. The higher values of Weibull's shape factor and scale parameter as corroborated by experimental findings indicated faster rate of SB release from PBAT/CMMT/SB nanocomposite film, when compared to the pristine PBAT film. Bacterial inhibition studies were accomplished against 2 food pathogenic bacteria, Bacillus subtilis and Staphylococcus aureus, by determining the zone of inhibition and corresponding growth profiles. Both bacterial inhibition studies and growth profiles established that PBAT/CMMT/SB demonstrated better antimicrobial activity than PBAT/SB film. Therefore, PBAT/CMMT/SB nanocomposite film can be used for food packaging application as it showed good barrier properties and antimicrobial activity against food pathogenic bacteria.

Process development and intensification for enhanced production of Bacillus lipopeptides.

The growing interest in Bacillus lipopeptides for high-value applications has driven process design, development and optimization for enhanced lipopeptide production. Traditional optimization approaches have been directed towards improving the overall titres by modification of media components and environmental parameters, almost exclusively in submerged cultures. Carbon and nitrogen sources, trace elements and oxygen availability have all been demonstrated to exhibit significant influences on lipopeptide yield, productivity and selectivity. This insight into process-linked kinetics, especially selectivity, has led to the introduction of novel process intensification and integration strategies which further promote process efficiency, and which include foam fractionation, inverse fluidization, rotating disc contacting and microfiltration with recycle. These strategies have not only transformed the production capabilities, but have also successfully integrated upstream production with downstream purification through cell retention and in situ product removal. This review analyses and critically discusses the impact of process conditions and process optimization strategies for improving lipopeptide production kinetics, specifically highlighting the emerging trend of process intensification and integration strategies and further, proposes a heuristic route to enhance lipopeptide production.

Potential therapeutic applications of biosurfactants.

Biosurfactants have recently emerged as promising molecules for their structural novelty, versatility, and diverse properties that are potentially useful for many therapeutic applications. Mainly due to their surface activity, these molecules interact with cell membranes of several organisms and/or with the surrounding environments, and thus can be viewed as potential cancer therapeutics or as constituents of drug delivery systems. Some types of microbial surfactants, such as lipopeptides and glycolipids, have been shown to selectively inhibit the proliferation of cancer cells and to disrupt cell membranes causing their lysis through apoptosis pathways. Moreover, biosurfactants as drug delivery vehicles offer commercially attractive and scientifically novel applications. This review covers the current state-of-the-art in biosurfactant research for therapeutic purposes, providing new directions towards the discovery and development of molecules with novel structures and diverse functions for advanced applications.

An inexpensive strategy for facilitated recovery of metals and fermentation products by foam fractionation process.

Microbial biosurfactants produce foam during aerobic-fermentation processes. The degree of foaminess and foam stability of the lipopeptide-biosurfactant produced by a marine Bacillus megaterium strain were investigated using simulated biosurfactant solution (SBS), biosurfactant broth without cells (BBWOC) and biosurfactant broth with cells (BBWC) in bubble column experiments. The experimental data for foam collapse were fitted using a first-order foam decay model. The first-order rate constant (k), a measure of foam stability, was maximum (k=0.0003 S(-1)) for BBWOC in the pH range 6-9. However, maximum foam stability (k=0.0006 S(-1)) was restricted to pH 7 for BBWC. Foam-based metal removal studies revealed that the metal removal followed a saturation model. The relative binding capacity of each divalent metal was greatly affected by the presence of other divalent metals. The order of lipopeptide binding capacity of the metals was Fe(2+)>Ca(2+)>Mg(2+), with Fe(2+) significantly influencing the foam stability. In case of Fe(2+), Ca(2+) and Mg(2+), maximum metal recovery of 64.7±4.3%, 52±3.1% and 41.4±3.6% respectively was observed at pH 7. The enrichment (E) of the other media components, including cells, was comparatively insignificant. The results of this study have implications in designing and optimizing biosurfactant or protein recovery in situ by foam fractionation as an inexpensive strategy, and also in facilitated metal recovery from industrial effluents and ores.