An antimicrobial metabolite n- hexadecenoic acid from marine sponge-associated bacteria Bacillus subtilis effectively inhibited biofilm forming multidrug-resistant P. aeruginosa.

Effective drug candidates to obstruct the emergence of multidrug-resistant pathogens have become a major concern. A potent antimicrobial producer was isolated from a marine sponge designated as MSI38 and was identified as Bacillus subtilis by 16SrDNA sequencing. The active antimicrobial fraction was purified, and the metabolite was identified as n-hexadecanoic acid by spectroscopic analysis. The fish-borne pathogen Pseudomonas aeruginosa FP012 was found to be multidrug-resistant and poses a risk of disease to food handlers and consumers in general. The compound showed a potent bactericidal effect against P. aeruginosa FP012 with a MIC of 31.33 ± 5.67 mg L-1 and MBC of 36.66 ± 5.17 mg L-1. The time-based biofilm inhibitory potential of MSI38 and ciprofloxacin was analyzed by confocal laser scanning microscopy. A synergistic effect of MSI38 and ciprofloxacin on biofilm showed 85% inhibition.

Effective utilization of wheat gluten by lipopeptide biosurfactant in the formulation of cookies.

BACKGROUND: Gluten-free food products are in demand due to the gluten sensitivity of individuals around the globe. Lipopeptide biosurfactants are widely used in food formulations for wetting, foaming, emulsion stabilization, anti-adhesive and anti-microbial properties. Lipopeptide biosurfactants can be used for gluten utilization and the formulation of food products. RESULTS: The strain Bacillus licheniformis MS48 was isolated from the marine sponge associated bacteria and found to be the ideal biosurfactant producer with an emulsification activity of 70%. Biosurfactant production was optimized using lactose, yeast extract, and sodium chloride (NaCl). The 1,1-diphenyl- 2-picryl hydrazyl (DPPH) radical scavenging activity was found to increase with an increase in the concentration of biosurfactant. The conformation changes in the gluten  due to the treatment of lipopeptide biosurfactant was identified by thiol quantification and scanning electron microscopy. With the addition of lipopeptide MS48 to cookie dough, cookies with softer and smooth textures were developed. The conformation changes in the gluten   in the lipopeptide incorporated  cookie dough was visualized by scanning electron microscopy. The lipopeptide biosurfactant incorporation in cookie dough causes appropriate changes in the structural network of gluten. The lipopeptide-incorporated cookies had improved the spread factor and texture. CONCLUSION: Lipopeptide biosurfactant with effective radical scavenging activity aided in the utilization of gluten. Incorporating lipopeptide in cookie dough assisted in the production of cookies with improved textural and sensory properties. This study is a novel approach to the application of lipopeptide biosurfactant in gluten utilization for the development of cookies. © 2023 Society of Chemical Industry.

Revealing the effect of lipopeptide on improving the probiotics characteristics: Flavor and texture enhancer in the formulated yogurt.

Probiotics are beneficial viable microorganisms that protects from pathogens, enhance immunity and health. In this study, a biosurfactant from marine sponge associated bacteria MS48 was used to effectively enhance the survival of starter cultures and thereby improving their functional properties. The biosurfactant MS48 was characterized as lipopeptide based upon the spectroscopic analysis. The GC-MS analysis showed the moiety as l-glutamic acid methyl ester with the mass of m/z 161.16. Probiotics supplemented with lipopeptide showed better survival in the stress tolerance assays includes acid, bile, heat, salt stress and other assays including auto-aggregation, hydrophobicity, microbial adhesion to solvents (MATS), and simulated gastric juice. Yogurt formulated using lipopeptide showed enhanced flavor components, stability, improved characteristics, EPS production, and lower syneresis than the control. Malonic acid, acidity regulator was detected in the mass spectrum of lipopeptide added yogurt. The texture analysis of the lipopeptide added yogurt showed improved textural and sensorial properties when compared to the control.

Anti-diabetic Potential of a Stigmasterol From the Seaweed Gelidium spinosum and Its Application in the Formulation of Nanoemulsion Conjugate for the Development of Functional Biscuits.

The seaweed Gelidium spinosum was selected for the extraction of phytosterol by the Soxhlet method. The extracted phytosterol was chemically characterized as stigmasterol using Fourier-transform infrared spectrometry and gas chromatography-mass spectrometry analysis. The antioxidant and α-amylase inhibitory activity of stigmasterol has been confirmed by in vitro assays. The in vivo studies demonstrated an anti-diabetic effect in streptozotocin (STZ)-induced hyperglycemic rats. Biochemical analysis showed administration of stigmasterol reduced the blood sugar, urea, and creatinine level. The stigmasterol was then nano-emulsified and incorporated into dough for biscuit formulation. The stigmasterol incorporated biscuit showed higher proximate values, low moisture content, lighter color and the textural property revealed lower hardness. Sensorial results showed acceptability when compared to the control. This study demonstrated the stigmasterol reduced hyperglycemic effects and therefore could be used as a supplement in diets for diabetic patients.

Revealing the Efficacy of Thermostable Biosurfactant in Heavy Metal Bioremediation and Surface Treatment in Vegetables.

Biosurfactants are amphiphilic molecules which showed application in the food, medical, and cosmetics industries and in bioremediation. In this study, a marine sponge-associated bacteria (MSI 54) was identified as a biosurfactant producer which showed high emulsification and surface tension-reducing property. The isolate MSI 54 was identified as Bacillus sp. and the biosurfactant was chemically characterized as a lipopeptide analog based on the spectral data including Fourier transform infrared spectroscopy and nuclear magnetic resonance spectroscopy. The MSI 54 lipopeptide biosurfactant was an anionic molecule which showed high affinity toward cationic heavy metals including Pb, Hg, Mn, and Cd. The heavy metal bioremediation efficacy of the biosurfactant was evaluated using atomic absorption spectroscopy, scanning electron microscopy/energy-dispersive X-ray spectroscopy, and high-resolution transmission electron microscopy analysis. When MSI 54 lipopeptide biosurfactant was added to heavy metals, this resulted in a white co-precipitate of the metal-biosurfactant complex. The heavy metal remediation efficacy of the biosurfactant at a 2.0 × critical micelle concentration (CMC) showed removal of 75.5% Hg, 97.73% Pb, 89.5% Mn, and 99.93% Cd, respectively, in 1,000 ppm of the respective metal solution. The surface treatment of farm fresh cabbage, carrot, and lettuce with 2.0 × CMC of the lipopeptide showed effective removal of the surface heavy metal contaminants.

Photoprotective effect of nanomelanin-seaweed concentrate in formulated cosmetic cream: With improved antioxidant and wound healing properties.

A melanin producer bacteria Halomonas venusta was isolated from a marine sponge Callyspongia sp. and optimized for melanin production. The optimized fermented media supplemented with 1% tyrosine yielded 4.92 mg/ml of melanin. The melanin incorporated cream was formulated and fortified with concentrates of seaweed Gelidium spinosum. Melanin and seaweed concentrate were found to be rich in antioxidant activity and were effectively inhibited the growth of S. aureus (MTCC 96) and S. pyogenes (MTCC 442). Various combinations of the cream were optimized and selected a formula containing 0.25% of melanin and 0.75% of seaweed concentrate which showed improved texture quality of cream. The formulated cream showed a pH of 5.52, spreadability 23 mm, and smooth and homogeneous texture. On application over skin provides a cooling effect and immediate disappearance without formation of white or oily film. Texture analysis of newly formulated cream showed similar results with that of control cream in terms of firmness, cohesiveness, index of viscosity and consistency. The formulated cream showed significant reduction of reactive oxygen species generated on exposure to direct sunlight. The cream showed protective effect on photohemolysis thus protecting the skin from lysis of red blood cells. The sun protection factor of the formulated cream F3 was found to be 18.373 ± 1.45. The combined antimicrobial and antioxidant effect of melanin and seaweed concentrate increased the shelf life of cream over the control. This study was the first report on photoprotective cream formulation using melanin and seaweed concentrate, which improved antioxidant and wound healing properties. The antimicrobial effect of the formulated natural cream could reduce the emergence of drug resistant bacteria and side effects of synthetic creams.

An antibiotic agent pyrrolo[1,2-a]pyrazine-1,4-dione,hexahydro isolated from a marine bacteria Bacillus tequilensis MSI45 effectively controls multi-drug resistant Staphylococcus aureus.

Sponge associated bacteria are a rich source of bioactive secondary metabolites. This study aims to isolate bacteria producing antimicrobial agents from a marine sponge, Callyspongia diffusa. A total of fifty-six bacteria were isolated and screened for antibacterial activity against multidrug resistant S. aureus. Based on the 16S rRNA sequence and phylogenetic analysis the antimicrobial producer strain MSI45 was identified as a novel Bacillus tequilensis. The culture conditions of strain MSI45 were optimized to enhance the yield of the antimicrobial compound. The antimicrobial compound was purified using a silica gel column chromatography and high performance liquid chromatography. On the basis of spectroscopic analysis such as FT-IR, NMR and GC-MS, the bioactive metabolite was identified as pyrrolo[1,2-a]pyrazine-1,4-dione,hexahydro. The extracted compound MSI45 showed a potent inhibitory effect on multidrug resistant S. aureus with an MIC of 15 ± 0.172 mg L-1 and MBC of 20 ± 0.072 mg L-1. The compound was non-hemolytic and showed high antioxidant activity. The antioxidant activity may increase the efficacy and safety of the molecule in drug development. Hence, this compound produced by Bacillus tequilensis MSI45 could have potent antimicrobial and antioxidant activity against S. aureus infection.