Isolation and characterization of two glycolipopeptids biosurfactants produced by a Lactiplantibacillus plantarum OL5 strain isolated from green olive curing water.

Microbial surfactants are natural amphiphilic compounds with high surface activities and emulsifying properties. Due to their structural diversity, low toxicity, biodegradability, and chemical stability in different conditions, these molecules are potential substitutes for chemical surfactants; their interest has grown significantly over the last decade. The current study focuses on the isolation, identification, and characterization of a lactic acid bacteria that produce two forms of biosurfactants. The OL5 strain was isolated from green olive fermentation and identified using MALDI/TOF and DNAr16S amplification. Emulsification activity and surface tension measurements were used to estimate biosurfactant production. The two biosurfactants derived from Lactiplantibacillus plantarum OL5 presented good emulsification powers in the presence of various oils. They were also shown to have the potential to reduce water surface tension from 69 mN/m to 34 mN/m and 37 mN/m within a critical micelle concentration (CMC) of 7 mg/ml and 1.8 mg/ml, respectively, for cell bound and extracellular biosurfactants. Thin layer chromatography (TLC) and FT-IR were used to analyze the composition of the two biosurfactants produced. the obtained data revealed that the two biomolecules consist of a mixture of carbohydrates, lipids and proteins. We demonstrated that they are two anionic biosurfactants with glycolipopeptide nature which are stable in extreme conditions of temperature, pH and salinity.

Bacillus subtilis SPB1 lipopeptide biosurfactant: antibacterial efficiency against the phytopathogenic bacteria Agrobacterium tumefaciens and compared production in submerged and solid state fermentation systems.

Bacillus subtilis SPB1 derived biosurfactants (BioS) proved its bio-control activity against Agrobacterium tumefaciens using tomato plant. Almost 83% of disease symptoms triggered by Agrobacterium tumefaciens were reduced. Aiming potential application, we studied lipopeptide cost-effective production in both fermentations systems, namely the submerged fermentation (SmF) and the solid-state fermentation (SSF) as well as the use of Aleppo pine waste and confectionery effluent as cheap substrates. Optimization studies using Box-Behnken (BB) design followed by the analysis with response surface methodology were applied. When using an effluent/sea water ratio of 1, Aleppo pine waste of 14.08 g/L and an inoculum size of 0.2, a best production yield of 17.16 ± 0.91 mg/g was obtained for the SmF. While for the SSF, the best production yield of 27.59 ± 1.63 mg/g was achieved when the value of Aleppo pine waste, moisture, and inoculum size were, respectively, equal to 25 g, 75%, and 0.2. Hence, this work demonstrated the superiority of SSF over SmF.

Optimization of textile effluent bacterial treatment and improvement of the process efficiency through SPB1 biosurfactant addition.

The current work aims to optimize biological textile effluent treatment through the use of newly selected bacterial consortia composed of two strains: Citrobacter sedlakii RI11 and Aeromonas veronii GRI. We assessed the effect of SPB1 biosurfactant addition on color removal (CR). The process was optimized by a Box-Bhenken by examining the effect of pH, consortia density and biosurfactant value on treatment efficiency. Firstly, physicochemical analyses of the studied effluent revealed an alkaline pH along with a high content of suspended materials and large amounts of organic matter. Optimal CR and a chemical oxygen demand abatement of about 94 and 86% were obtained when treating the textile effluent at pH 5 with a total optical density of 0.4 and by incorporating 0.01% SPB1 biosurfactant. Additionally, an abolishment of phytotoxicity was registered after treatment optimization. The evaluations of the action mode of both selected bacteria during textile effluent treatment suggested the occurrence of biodegradation phenomena of dyes through enzymatic activities.

Enhanced biosurfactant production by Bacillus subtilis SPB1 using developed fed-batch fermentation: effects of glucose levels and feeding systems.

Biosurfactants stand for highly useful and promising compounds. They basically serve for a variety of applications in multiple industries and aspects of human life. Therefore, it is highly required to improve their production yield especially through the development of new and more efficient fermentation processes. In this aim, batch and fed-batch were studied and compared in terms of their effective biosurfactant production by Bacillus subtilis SPB1. Experiments of fed-batch fermentations were carried out through three different glucose feeding strategies, namely the pulsed, the constant Donespeed and the exponential feeding. The comparison between different fermentation processes revealed that fed-batch process proved to be a more efficient cultivation strategy than the batch process in terms of cell biomass, biosurfactant production and productivity. Among the three different feeding strategies, the exponential feeding process achieved the highest fermentation results of final biosurfactant concentration. The latter increased more than twofolds compared to batch fermentation.

Surface properties and heavy metals chelation of lipopeptides biosurfactants produced from date flour by Bacillus subtilis ZNI5: optimized production for application in bioremediation.

The present study summarizes the valorization of date flour by the production of lipopeptide biosurfactant (BioS) by Bacillus subtilis ZNI5 (MW091416). A Taguchi design permitted the formulation of a medium composed only of 6% date flour and 0.5% yeast extract within 2 days of incubation at 150 rpm with a maximal surface tension (ST) reduction of about 27.8 mN/m. The characterization of the lipopeptide shows a CMC value of about 400 mg/L with a minimal ST of 30 mN/m and an ability to disperse oil to about 80 mm at 800 mg/L. Having reduced phytotoxicity, the ZNI5 BioS and ZNI5 strain were assayed for Copper and Cobalt chelation and biosorption. The improvement of the germination index of radish seeds irrigated by the treated contaminated water showed the great potential application of ZNI5 lipopeptide in the bioremediation of heavy metals.

Potential application of Bacillus subtilis SPB1 biosurfactants in laundry detergent formulations: Compatibility study with detergent ingredients and washing performance.

Surfactants play a very important role in laundry and household cleaning products ingredients. In this research, the application of lipopeptide biosurfactants, produced by Bacillus subtilis SPB1, in the formulation of a washing powder was investigated. The SPB1 biosurfactant was mixed with sodium tripolyphosphate as a builder and sodium sulfate as filler. The efficiency of the formulated detergent composition with different washing conditions to remove a stain from cotton fabric was examined. The results showed that the formulated detergent was effective in oil removal, with optimal washing conditions of pH, temperature, striate and time of washing system of 7, 65°C, 1000 RPM and 60 min, respectively. A comparative study of different detergent compositions (biosurfactant-based detergent, combined biosurfactant-commercial detergent, and a commercial detergent) for the removal of oil and tea stains, proved that the bio-scouring was more effective (>75%) in terms of the stain removal than the commercial powders (<60%). Moreover, the results demonstrated that the biosurfactant acts additively with a commercial detergent and enhances their performance from 33 to 45% in removing oil stain and from 57 to 64% in removing tea stain. As a conclusion, in addition to the low toxicity and the high biodegradability of the microbial biosurfactants, the results of this study have shown that the future use of this lipopeptide biosurfactant as laundry detergent additive is highly promising.

Improved Biosurfactant Production by Bacillus subtilis SPB1 Mutant Obtained by Random Mutagenesis and Its Application in Enhanced Oil Recovery in a Sand System.

Biosurfactants or microbial surfactants are surface-active biomolecules that are produced by a variety of microorganisms. Biodegradability and low toxicity have led to the intensification of scientific studies on a wide range of industrial applications for biosurfactants in the field of environmental bioremediation as well as the petroleum industry and enhanced oil recovery. However, the major issues in biosurfactant production are high production cost and low yield. Improving the bioindustrial production processes relies on many strategies, such as the use of cheap raw materials, the optimization of medium-culture conditions, and selecting hyperproducing strains. The present work aims to obtain a mutant with higher biosurfactant production through applying mutagenesis on Bacillus subtilis SPB1 using a combination of UV irradiation and nitrous acid treatment. Following mutagenesis and screening on blood agar and subsequent formation of halos, the mutated strains were examined for emulsifying activity of their culture broth. A mutant designated B. subtilis M2 was selected as it produced biosurfactant at twice higher concentration than the parent strain. The potential of this biosurfactant for industrial uses was shown by studying its stability to environmental stresses such as pH and temperature and its applicability in the oil recovery process. It was practically stable at high temperature and at a wide range of pH, and it recovered above 90% of motor oil adsorbed to a sand sample.

Potential application of Bacillus subtilis SPB1 lipopeptides in toothpaste formulation.

Toothpaste is a gel dentifrice used with a toothbrush as an accessory to clean, keep and promote oral hygiene. The literature review suggests that there are many different formulations of toothpastes and that each of their individual components present specific functions. The concentration of the toothpaste ingredients must be appropriately chosen taking into account the purposes of the toothpaste. Biosurfactants are considered as suitable molecules for application in many formulations such as in toothpaste one. In the present work, two dentifrice formulations were investigated and their efficiencies were tested using chemical surfactant agent and lipopeptide biosurfactant isolated from Bacillus subtilis SPB1. The physicochemical properties were analyzed considering several tests mainly spreading ability, water activity, pH, foaming and cleaning tests. The obtained results indicated that the SPB1 biosurfactant was as efficient as the chemical surfactant confirming its potential utilization in toothpaste formulation compared to the commercial one. The evaluation of the antimicrobial activity of the formulated dentifrice was carried out against eight bacteria. The results demonstrated that the biosurfactant-based product exhibited an important antimicrobial activity, which was very effective against Enterobacter sp and Salmonella typhinirium.

Anionic lipopeptides from Bacillus mojavensis I4 as effective antihypertensive agents: Production, characterization, and identification.

A new isolated Bacillus mojavensis strain I4 was found as producer of biosurfactants by different screening methods, such as parafilm M test, hemolytic activity, oil displacement test, emulsification index, surface tension, and lipase production assay. Enhanced biosurfactants production was obtained using glucose and glutamic acid as carbon and nitrogen sources, respectively. The optimal production of the biosurfactants was obtained by using a C/N ratio of 17, pH of 7.0, and temperature of 37°C. The surface tension was reduced to 29 mN/m and the emulsification index E24 of 62% was achieved after 72 h of culture. The purified biosurfactants showed stability with regard to surface tension reduction and emulsification in a wide range of temperatures (4-120°C), pH (4-10), and salinity (2-12% of NaCl). The thin-layer chromatography showed that the produced biosurfactants were lipopeptides. The biosurfactants were characterized as a group of anionic lipopeptides with zeta potential measurement. Chromatographic characterization using HPLC revealed that I4 lipopeptides contained numerous isoforms and surfactin was the major component. Moreover, the I4 lipopeptides showed interesting angiotensin-converting enzyme-inhibitory activity.