First report of betalain production from endolichenic Bacillus sp. LDAB-1 from Dirinaria aegilita: Insights from novel quantification methodology of image processing.

Pigments are widely used in food supplements envisaging attractive colors along with health benefits. The desired advancements in the nutraceutical and antioxidant properties of pigments utilized in food products necessitate the search for novel additives. The present study is the first in the field to report the pigment-producing endolichenic bacteria, Bacillus sp. LDAB-1 from Dirinaria aegilita. Morphological, biochemical, and molecular characterization of the bacterium emphasizes that ideal pigment production occurs when utilizing sucrose and sodium nitrate. The pigment was salted out and dialyzed for further qualitative characterization using ultraviolet-visible, fluorescence, and Fourier transform infrared spectra and the results corroborated the presence of betalains. The antioxidant activity of betalain is closer to the efficiency of α-tocopherol, which confers the pigment properties for antioxidant and nutraceutical significance. An optimal methodology for pigment affirmation is an issue when using an alternative methodology. Hence, the present assessment employs a comparative analysis of findings from both a spectrophotometric method and image processing technology encompassing RGB, CMYK, YCbCr, and L*a*b* color space models. Amongst these, the L*a*b* model potentially provides an effective modality for determining the pigment concentration. Bland-Altman plot analysis indicates similar consistency levels in betalain quantification by both methods at 95% confidence intervals, affirming the integrity and consistency of color image processing technology. Consequently, the present study represents novelty and innovativeness in reporting endolichenic Bacillus sp. LDAB-1 from D. aegilita and a rational image optimization protocol for pigment elucidation characteristics.

Unveiling the impact of additives on structural integrity, thermal and color stability of C-phycocyanin - Agar hydrocolloid.

C-Phycocyanin and sugar (C-PC/S) blended agar hydrocolloid was prepared and its rheological, thermo-functional and morphological properties were examined based on the fluorescence excitation-emission matrix profile. Sucrose (40%, w/v) determined as a superior preservative, maintaining the native conformation of C-PC effectively. C-PC/S exhibited enhanced structural integrity with high storage modulus (G') and 86.4% swelling index. FT-IR demonstrated strong intramolecular bonding. TGA revealed that the presence of sucrose prolonged the devolatilization peak up to 325 °C, with a degradation rate of -2.273 mg/min, it the thermal stability. C-PC/S fortified hydrocolloid in ice cream (5.0% w/w), reduced melting rate up to five times. In conclusion, sucrose as a promising enhancer of color stability and structural integrity for C-PC, and this combination effectively improves the functional and rheological properties. Further, the findings exposed the agar hydrocolloid as a potential enhancer of color retention and improved performance for various food and cosmetic products.

Development and Characterization of Bio-Based Composite Films for Food Packing Applications Using Boiled Rice Water and Pistacia vera Shells.

Customer demand for natural packaging materials in the food industry has increased. Biocomposite films developed using boiled rice water could be an eco-friendly and cost-effective packaging product in the future. This study reports the development of bio-based films using waste materials, such as boiled rice water (matrix) and Pistacia vera shells (reinforcement material), using an adapted solution casting method. Several film combinations were developed using various concentrations of plasticizing agent (sorbitol), thickening agent (oil and agar), and stabilizing agents (Arabic gum, corn starch, and Pistacia vera shell powder). Various packaging properties of the film were analyzed and examined to select the best bio-based film for food packaging applications. The film fabricated with Pistacia vera shell powder in the biocomposite film exhibited a reduced water solubility, swelling index, and moisture content, as compared to polyethene packaging material, whereas the biocomposite film exhibited poor antimicrobial properties, high vapor transmission rate, and high biodegradability rate. The packaging properties and characterization of the film indicated that the boiled rice water film with Pistacia vera shell powder was suitable for packaging material applications.

Characterization and antifungal activity of the yellow pigment produced by a Bacillus sp. DBS4 isolated from the lichen Dirinaria agealita.

This study emphasis the production of yellow pigment from endolichenic Bacillus sp. isolated from the lichen Dirinaria aegialita (Afzel. ex Ach.) B.J. Moore. Yellow pigment-producing twenty different strains were investigated. The hyperactive pigment-producing bacterial strain was identified as Bacillus gibsonii based on 99 % sequence similarity. Maximum bacterial pigment production appeared in Luria Bertani medium. Methanol extraction of the pigment and its partial purification using TLC was carried out. Furthermore, isolated pigments were characterized using UV-visible spectroscopy, FTIR spectroscopy, and GC-MS results related to the possibility of the carotenoid occurrence. The pigment also exhibited efficient antifungal activity against selected fungal pathogens of economic importance. Likewise, the pigment extract evaluated for the total antioxidant potential using Phosphomolybdenum and Ferric reducing antioxidant power assay and the results represented in Ascorbic Acid Equivalent (AAE)- 21.45 ± 1.212 mg/mL. The SC50 of the pigment extract found to be 75.125 ± 0.18 µg/ml determined by the ABTS assay.

Digital image-based quantification of chlorpyrifos in water samples using a lipase embedded paper based device.

A paper-based device (PBD) for the detection of chlorpyrifos pesticide at field application was fabricated based on the principles of enzyme inhibition and image processing. Rhizopus niveus lipase, p-nitrophenol palmitate and Whatman No.1 paper were used as an enzyme, substrate and support matrix, respectively. The performance of functionalized PBD was tested for lateral flow assay reaction in pure water (negative control), artificial pesticide water (positive control) and selected fruits and vegetables wash water (test). The digital image of the PBD after the test was captured using an android smartphone and analyzed in MATLAB software. Different colour space models such as, grey, RGB, HSV and YCbCr were studied and the Cb coordinate was chosen for its higher linearity (R2 = 0.988) with pesticide concentration. Experimental variations such as paper length, relative concentration ratio of the substrate and enzyme were investigated to minimize the product cost and analysis time. The developed PBD showed a significant response over wide range of sample solution's pH and operational temperature. Further, a long-term storage stability was measured for developed PBD. The LOD and LOQ were found to be 0.065 mgL-1 and 0.198 mgL-1. The results obtained from newly developed image processing method showed 92.8% accuracy with microtiter plate assay. Higher MRL was determined in the wash water of cauliflower, grapes, coriander leaves, brinjal and bitter guard. Overall, the developed paper biosensor was precise, cost effective and most suitable for field applications.

Synthesis of Feruloyl Ester Using Bacillus subtilis AKL 13 Lipase Immobilized on Celite® 545.

The lipophilic antioxidants, glyceryl ferulate and feruloyl glyceryl linoleate, were synthesized using lipase from Bacillus subtilis AKL 13. The extracellular lipase was produced by cultivation of the strain in modified minimal medium and the enzyme was recovered by fractionation at 80% ammonium salt saturation. The concentrated enzyme with the specific activity of (4647±66) U/mg was immobilized on Celite® 545 and crosslinked using glutaraldehyde. The prepared enzyme catalyst was used for esterification of ferulic and linoleic acids with glycerol separately in hexane butane solvent system at 50 °C and 3.144×g agitation. The maximum ester conversion of 94% of feruloyl glyceryl linoleate was achieved at 48 h, whereas only 35% of glyceryl ferulate was synthesized. The reaction products were characterized using RP-HPLC, FTIR, 1H NMR, 13C NMR and fluorescence spectrophotometry. The kinetic parameters of esterification reaction were determined according to ping-pong bi-bi model. The Km and υmax were found to be 69.37 and 3.46 mmol, and 0.387 and 1.02 mmol/(min·g) for glyceryl ferulate and feruloyl glyceryl linoleate, respectively. The kinetic parameters were simulated in MATLAB and the experimental data were in good agreement. Furthermore, 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity of the blend of feruloyl ester and palm oil was higher than of the plain palm oil and was closer to α-tocopherol.