Inhibition of pathogenic bacterial biofilms on PDMS based implants by L. acidophilus derived biosurfactant

Abstract Background Lactobacillus spp. predominantly shows its presence as a normal mucosal flora of the mouth and intestine. Therefore, the objective of our research is to investigate the in-vitro conditions for the prospective of medically valuable biosurfactants (BSs) derived from Lactobacillus spp. Biosurfactant (BS) obtained from Lactobacillus spp. exhibit antibiofilm and antiadhesive activity against broad range of microbes. In the present study we investigated the production, purification and properties of key components of the cell-associated-biosurfactant (CABS) from Lactobacillus acidophilus NCIM 2903. Results Extracted, purified, freeze-dried CABS shows reduction in surface tension (SFT) of phosphate buffer saline (PBS @pH 7.0) from 71 to 26 mN/m and had a critical micelle concentration (CMC) of 23.6 mg/mL. The CABS showed reduction in interfacial tension (IFT) against various hydrocarbons and had effective spreading capability as reflected through the decrease in contact angle (CA) on different surfaces (polydimethylsiloxane - PDMS, Teflon tape, glass surface, polystyrene film and OHP sheet). The anionic nature of CABS displayed stability at different pH and temperatures and formed stable emulsions. Thin layer chromatography (TLC) and Fourier transform infrared spectroscopy (FTIR) revealed CABS as glycolipoprotein type. The Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE) showed presence of multiple bands in a molecular range of 14.4 to 60 kDa, with prominent bands of 45 kDa. The CABS has significant antiadhesion and antibiofilm activity against tested bacterial strains. Conclusion The current challenging situation is to develop methods or search for the molecules that will prevent the formations of biofilm on medical bioimplants of PDMS based materials. These findings are supportive for the use of Lactobacilli derived BS as potential antiadhesive agent on various surfaces of biomedical devices

Lactobacillus acidophilus Derived Biosurfactant as a Biofilm Inhibitor: A Promising Investigation Using Microfluidic Approach

Background: Biomedical devices and implants are adversely affected by biofilm-associated infections that pose serious public health issues. Biosurfactants (BSs) can combat pathogenic biofilms through their antimicrobial, antibiofilm and antiadhesive capabilities. The objective of our research was to produce biosurfactant (BS) from Lactobacillus acidophilus NCIM 2903 and investigate its antibiofilm, antiadhesive potential using microfluidics strategies by mimicking the micro-environment of biofilm. Methods: Antibiofilm and antiadhesive potential was effectively evaluated using different methods like microfluidics assay, catheter assay, polydimethlysiloxane (PDMS) disc assay. Along with this chemical and physical characteristics of BS were also evaluated. Results: Cell free biosurfactant (CFBS) obtained was found to be effective against biofilm which was validated through the microfluidic (MF) or Lab on Chip (LOC) approach. The potency of CFBS was also evaluated on catheter tubing and PDMS surfaces (representative bioimplants). The efficacy of CFBS was also demonstrated through the reduction in surface tension, interfacial tension, contact angle and low critical micelle concentration. Conclusion: CFBS was found to be a potent antimicrobial and antibiofilm agent. We believe that perhaps this is the first report on demonstrating the inhibiting effect of Lactobacillus spp. derived CFBS against selected bacteria via LOC approach. These findings can be explored to design various BSs based formulations exhibiting antimicrobial, antibiofilm and antiadhesive potential for biomedical applications

Genomic Insights of Halophilic Planococcus maritimus SAMP MCC 3013 and Detail Investigation of Its Biosurfactant Production

Moderate halophilic bacteria thrive in saline conditions and produce biosurfactant (BS) which facilitates the oil scavenging activity in the oil polluted surroundings. Production of such unusual bioactive molecules plays a vital role for their survival in an extreme and adverse environment. Current research deals with isolation of Planococcus maritimus strain SAMP MCC 3013 from Indian Arabian coastline sea water for BS production. The bacterium tolerated up to 2.7 M NaCl demonstrating osmotic stress bearable physiological systems. We used integrated approach to explore the genomic insight of the strain SAMP and displayed the presence of gene for BS biosynthesis. The genome analysis revealed this potential to be intrinsic to the strain. Preliminary screening techniques viz., surface tension (SFT), drop collapse (DC) and oil displacement (OD) showed SAMP MCC 3013 as a potent BS producer. BS reduced SFT of phosphate buffer saline (PBS) pH: 7.0 from 72 to 30 mN/m with a critical micelle concentration (CMC) value of 1.3 mg/mL. Subsequent investigation on chemical characterization, using thin layer chromatography (TLC), Fourier transform infrared spectroscopy (FT-IR), nuclear magnetic resonance (1H NMR and 13C NMR) and liquid chromatography mass spectrometry (LC-MS) revealed terpene containing BS having sugar, lipid moieties. The genomic sequence analysis of P. maritimus SAMP showed complete genes in the pathway for the synthesis of terpenoid. Probably terpenoid is the accountable backbone molecule for the BS production, but the later stages of terpenoid conversion to the BS could not be found. Moreover, it is important to highlight that till today; no single report documents the in-detailed physico-chemical characterization of BS from Planococcus sp. Based on genomic and functional properties, the term terpene containing BS is denoted for the surfactant produced by P. maritimus

Biosurfactants’ multifarious functional potential for sustainable agricultural practices

Increasing food demand by the ever-growing population imposes an extra burden on the agricultural and food industries. Chemical-based pesticides, fungicides, fertilizers, and high-breeding crop varieties are typically employed to enhance crop productivity. Overexploitation of chemicals and their persistence in the environment, however, has detrimental effects on soil, water, and air which consequently disturb the food chain and the ecosystem. The lower aqueous solubility and higher hydrophobicity of agrochemicals, pesticides, metals, and hydrocarbons allow them to adhere to soil particles and, therefore, continue in the environment. Chemical pesticides, viz., organophosphate, organochlorine, and carbamate, are used regularly to protect agriculture produce. Hydrophobic pollutants strongly adhered to soil particles can be solubilized or desorbed through the usage of biosurfactant/s (BSs) or BS-producing and pesticide-degrading microorganisms. Among different types of BSs, rhamnolipids (RL), surfactin, mannosylerythritol lipids (MELs), and sophorolipids (SL) have been explored extensively due to their broad-spectrum antimicrobial activities against several phytopathogens. Different isoforms of lipopeptide, viz., iturin, fengycin, and surfactin, have also been reported against phytopathogens. The key role of BSs in designing and developing biopesticide formulations is to protect crops and our environment. Various functional properties such as wetting, spreading, penetration ability, and retention period are improved in surfactant-based formulations. This review emphasizes the use of diverse types of BSs and their source microorganisms to challenge phytopathogens. Extensive efforts seem to be focused on discovering the innovative antimicrobial potential of BSs to combat phytopathogens. We discussed the effectiveness of BSs in solubilizing pesticides to reduce their toxicity and contamination effects in the soil environment. Thus, we have shed some light on the use of BSs as an alternative to chemical pesticides and other agrochemicals as sparse literature discusses their interactions with pesticides. Life cycle assessment (LCA) and life cycle sustainability analysis (LCSA) quantifying their impact on human activities/interventions are also included. Nanoencapsulation of pesticide formulations is an innovative approach in minimizing pesticide doses and ultimately reducing their direct exposures to humans and animals. Some of the established big players and new entrants in the global BS market are providing promising solutions for agricultural practices. In conclusion, a better understanding of the role of BSs in pesticide solubilization and/or degradation by microorganisms represents a valuable approach to reducing their negative impact and maintaining sustainable agricultural practices

Synergistic Activity of Rhamnolipid Biosurfactant and Nanoparticles Synthesized Using Fungal Origin Chitosan Against Phytopathogens

Phytopathogens pose severe implications in the quantity and quality of food production by instigating several diseases. Biocontrol strategies comprising the application of biomaterials have offered endless opportunities for sustainable agriculture. We explored multifarious potentials of rhamnolipid-BS (RH-BS: commercial), fungal chitosan (FCH), and FCH-derived nanoparticles (FCHNPs). The high-quality FCH was extracted from Cunninghamella echinulata NCIM 691 followed by the synthesis of FCHNPs. Both, FCH and FCHNPs were characterized by UV-visible spectroscopy, DLS, zeta potential, FTIR, SEM, and Nanoparticle Tracking Analysis (NTA). The commercial chitosan (CH) and synthesized chitosan nanoparticles (CHNPs) were used along with test compounds (FCH and FCHNPs). SEM analysis revealed the spherical shape of the nanomaterials (CHNPs and FCHNPs). NTA provided high-resolution visual validation of particle size distribution for CHNPs (256.33 ± 18.80 nm) and FCHNPs (144.33 ± 10.20 nm). The antibacterial and antifungal assays conducted for RH-BS, FCH, and FCHNPs were supportive to propose their efficacies against phytopathogens. The lower MIC of RH-BS (256 μg/ml) was observed than that of FCH and FCHNPs (>1,024 μg/ml) against Xanthomonas campestris NCIM 5028, whereas a combination study of RH-BS with FCHNPs showed a reduction in MIC up to 128 and 4 μg/ml, respectively, indicating their synergistic activity. The other combination of RH-BS with FCH resulted in an additive effect reducing MIC up to 128 and 256 μg/ml, respectively. Microdilution plate assay conducted for three test compounds demonstrated inhibition of fungi, FI: Fusarium moniliforme ITCC 191, FII: Fusarium moniliforme ITCC 4432, and FIII: Fusarium graminearum ITCC 5334 (at 0.015% and 0.020% concentration). Furthermore, potency of test compounds performed through the in vitro model (poisoned food technique) displayed dose-dependent (0.005%, 0.010%, 0.015%, and 0.020% w/v) antifungal activity. Moreover, RH-BS and FCHNPs inhibited spore germination (61–90%) of the same fungi. Our efforts toward utilizing the combination of RH-BS with FCHNPs are significant to develop eco-friendly, low cytotoxic formulations in future

Lactobacillus acidophilus Derived Biosurfactant as a Biofilm Inhibitor: A Promising Investigation Using Microfluidic Approach

Background: Biomedical devices and implants are adversely affected by biofilm-associated infections that pose serious public health issues. Biosurfactants (BSs) can combat pathogenic biofilms through their antimicrobial, antibiofilm and antiadhesive capabilities. The objective of our research was to produce biosurfactant (BS) from Lactobacillus acidophilus NCIM 2903 and investigate its antibiofilm, antiadhesive potential using microfluidics strategies by mimicking the micro-environment of biofilm. Methods: Antibiofilm and antiadhesive potential was effectively evaluated using different methods like microfluidics assay, catheter assay, polydimethlysiloxane (PDMS) disc assay. Along with this chemical and physical characteristics of BS were also evaluated. Results: Cell free biosurfactant (CFBS) obtained was found to be effective against biofilm which was validated through the microfluidic (MF) or Lab on Chip (LOC) approach. The potency of CFBS was also evaluated on catheter tubing and PDMS surfaces (representative bioimplants). The efficacy of CFBS was also demonstrated through the reduction in surface tension, interfacial tension, contact angle and low critical micelle concentration. Conclusion: CFBS was found to be a potent antimicrobial and antibiofilm agent. We believe that perhaps this is the first report on demonstrating the inhibiting effect of Lactobacillus spp. derived CFBS against selected bacteria via LOC approach. These findings can be explored to design various BSs based formulations exhibiting antimicrobial, antibiofilm and antiadhesive potential for biomedical applications

BIOSURFACTANTS’ PRODUCTION FROM RENEWABLE NATURAL RESOURCES: EXAMPLE OF INNOVATIVEAND SMART TECHNOLOGY IN CIRCULAR BIOECONOMY

A strong developed bio-based industrial sector will significantly reduce dependency on fossil resources, help the coun-tries meet climate change targets, and lead to greener and more environmental friendly growth. The key is to develop new technologies to sustainably transform renewable natural resources into bio-based products and biofuels. Biomass is a valuable resource and many parameters need to be taken in to account when assessing its use and the products made from its. The bioeconomy encompass the production of renewable biological resources and their conversion into food, feed and bio-based products (chemicals, materials and fuels) via innovative and efficient technologies provided by indus-trial biotechnology. The paper presents the smart and efficient way to use the agro-industrial, dairy and food processing wastes for biosurfactant’s production. Clarification processes are mandatory to use the raw substrates for microbial growth as well as biosurfactant production for commercial purposes. At the same time it is very essential to retain the nutritional values of those cheap substrates. Broad industrial perspectives can be achieved when quality as well as the quantity of the biosurfactant is considered in great depth. Since substrates resulting from food processing, dairy, animal fat industries are not explored in great details; and hence are potential areas which can be explored thoroughly

Biosurfactants’ Production from Renewable Natural Resources: Example of Innovativeand Smart Technology in Circular Bioeconomy

A strong developed bio-based industrial sector will significantly reduce dependency on fossil resources, help the coun-tries meet climate change targets, and lead to greener and more environmental friendly growth. The key is to develop new technologies to sustainably transform renewable natural resources into bio-based products and biofuels. Biomass is a valuable resource and many parameters need to be taken in to account when assessing its use and the products made from its. The bioeconomy encompass the production of renewable biological resources and their conversion into food, feed and bio-based products (chemicals, materials and fuels) via innovative and efficient technologies provided by indus-trial biotechnology. The paper presents the smart and efficient way to use the agro-industrial, dairy and food processing wastes for biosurfactant’s production. Clarification processes are mandatory to use the raw substrates for microbial growth as well as biosurfactant production for commercial purposes. At the same time it is very essential to retain the nutritional values of those cheap substrates. Broad industrial perspectives can be achieved when quality as well as the quantity of the biosurfactant is considered in great depth. Since substrates resulting from food processing, dairy, animal fat industries are not explored in great details; and hence are potential areas which can be explored thoroughly