Optimisation of multigrain seera from sorghum, green gram and finger millet: effect of ingredients on functional, structural and thermal properties.

Seera, is a traditional Indian fermented food, has high carbohydrate and reducing sugar content, however, lacks functional components like antioxidant properties. The present study aims to optimize and evaluate the multigrain seera with added values. The optimization of seera was made using mixture design, with green gram (X1, 15-30%), sorghum (X2, 15-25%) and finger millet (X3, 5-10%) as independent variables. The responses checked were total phenolic content (Y1), protein content (Y2) and overall acceptability (Y3). The optimum run with green gram (25.58%), sorghum (15%) and finger millet (9.41%) resulted in TPC (1.2 ± 0.12 mg GAE/g), protein content (11.40 ± 0.10 g) with overall acceptability (8.32 ± 0.30). The optimized multigrain seera depicted higher fibre (4.23 ± 0.08%), ash (1.90 ± 1.1%) and protein (11.40 ± 0.10%) than the control seera. The rheological properties of seera depicted shear thinning and elastic behaviour. Texture profile analysis showed that cohesiveness (0.415 ± 0.01) increased significantly (along with decreased springiness (0.251). Morphology of seera showed broken and deformed starch granules with few cracks due to fermentation phenomena that leads to superficial corrosion. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-023-05854-5.

Isolation and optimization of extracellular PHB depolymerase producer Aeromonas caviae Kuk1-(34) for sustainable solid waste management of biodegradable polymers.

Bioplastics, synthesized by several microbes, accumulates inside cells under stress conditions as a storage material. Several microbial enzymes play a crucial role in their degradation. This research was carried to test the biodegradability of poly-ß-hydroxybutyrate (PHB) utilizing PHB depolymerase, produced by bacteria isolated from sewage waste soil samples. Potent PHB degrader was screened based on the highest zone of hydrolysis followed by PHB depolymerase activity. Soil burial method was employed to check their degradation ability at different incubation periods of 15, 30, and 45 days at 37±2°C, pH 7.0 at 60% moisture with 1% microbial inoculum of Aeromonas caviae Kuk1-(34) (MN414252). Without optimized conditions, 85.76% of the total weight of the PHB film was degraded after 45 days. This degradation was confirmed with Fourier-transform infrared spectroscopy (FTIR) and Scanning electron microscope (SEM) analysis. The presence of bacterial colonies on the surface of the degraded film, along with crest, holes, surface erosion, and roughness, were visible. Media optimization was carried out in statistical mode using Plackett Burman (PB) and Central Composite Design (CCD) of Response Surface Methodology (RSM) by considering ten different factors. Analysis of Variance (ANOVA), Pareto chart, response surface plots, and F-value of 3.82 implies that the above statistical model was significant. The best production of PHB depolymerase enzyme (14.98 U/mL) was observed when strain Kuk1-(34) was grown in a media containing 0.1% PHB, K2HPO4 (1.6 gm/L) at 27 ℃ for seven days. Exploiting these statistically optimized conditions, the culture was found to be a suitable candidate for the management of solid waste, where 94.4% of the total weight of the PHB film was degraded after 45 days of incubation.

Anti-Adhesion and Antibiofilm Activity of Eruca sativa Miller Extract Targeting Cell Adhesion Proteins of Food-Borne Bacteria as a Potential Mechanism: Combined In Vitro-In Silico Approach.

Bacterial cells have the ability to form biofilm onto the surfaces of food matrixes and on food processing equipment, leading to a source of food contamination posing serious health implications. Therefore, our study aimed to determine the effect of Eruca sativa Miller (E. sativa) crude extract against biofilms of food-borne bacteria along with in silico approaches to investigate adhesion proteins responsible for biofilm activity against the identified phytochemicals. The antibacterial potential of crude extract was evaluated using agar well diffusion technique and combinations of light and scanning electron microscopy to assess the efficacy of crude extract against the developed biofilms. Our results showed that crude extract of E. sativa was active against all tested food-borne bacteria, exhibiting a rapid kinetics of killing bacteria in a time-dependent manner. MIC and MBC values of E. sativa crude extract were found to be ranging from 125 to 500 µg/mL and 250 to 1000 µg/mL respectively. Furthermore, inhibition of developed biofilm by E sativa was found to be ranging from 58.68% to 73.45% for all the tested strains. The crude extract also reduced the viability of bacterial cells within biofilms and amount of EPS (ranging 59.73-82.77%) in the biofilm matrix. Additionally, the microscopic images also revealed significant disruption in the structure of biofilms. A molecular docking analysis of E. sativa phytochemicals showed interaction with active site of adhesion proteins Sortase A, EspA, OprD, and type IV b pilin of S. aureus, E. coli, P. aeruginosa, and S. enterica ser. typhi, respectively. Thus, our findings represent the first demonstration of E. sativa crude extract's bioactivity and potency against food-borne bacteria in their planktonic forms, as well as against the developed biofilms. Therefore, a possible mechanistic approach for inhibition of biofilm via targeting adhesion proteins can be explored further to target biofilm producing food-borne bacterial pathogens.

Real Time-PCR coupled with melt curve analysis for detecting the authenticity of camel milk.

The study evaluated the use of Real Time-Polymerase Chain Reaction (RT- PCR) to detect the adulteration of camel milk with goat, cow milk. DNA was isolated from camel milk, camel milk powder, camel milk soap, cow milk, and goat milk using DNA extraction kit. RT- PCR amplified a single piece of DNA into millions of copies. The camel specific primers were designed using the primer- 3 online software and quantification of the isolated DNA was carried out by RT- PCR system through DNA standard curves and cycle threshold (Ct) values. The detection limit of DNA template was in the range of 0.001-0.002%. The reaction mixture (20µL) contained 10 µL SYBR Green master mix, 0.3 µL of 10 µM of each primer and 5 µL DNA. Thermal cycling consisted of an initial denaturation at 95 °C for 1 min, followed by 40 cycles for 15 s at 95 °C and 60 °C for 30 s. The primer pairs used were confirmed for their PCR efficiency, and specific products were evaluated by melt curve analysis. Results indicated positive amplification for the camel milk, camel milk powder, and camel milk soap but negative amplification for cow and goat milk. In conclusion, the RT- PCR based identification is a low cost and appropriate method for camel milk and its products. Although, the yield of DNA from camel milk soap after isolation is low but the isolated DNA segment was easily identified.

Phytochemical and In Silico ADME/Tox Analysis of Eruca sativa Extract with Antioxidant, Antibacterial and Anticancer Potential against Caco-2 and HCT-116 Colorectal Carcinoma Cell Lines.

Eruca sativa Mill. (E. sativa) leaves recently grabbed the attention of scientific communities around the world due to its potent bioactivity. Therefore, the present study investigates the metabolite profiling of the ethanolic crude extract of E. sativa leaves using high resolution-liquid chromatography-mass spectrometry (HR-LC/MS), including antibacterial, antioxidant and anticancer potential against human colorectal carcinoma cell lines. In addition, computer-aided analysis was performed for determining the pharmacokinetic properties and toxicity prediction of the identified compounds. Our results show that E. sativa contains several bioactive compounds, such as vitamins, fatty acids, alkaloids, flavonoids, terpenoids and phenols. Furthermore, the antibacterial assay of E. sativa extract showed inhibitory effects of the tested pathogenic bacterial strains. Moreover, the antioxidant activity of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and hydrogen peroxide (H2O2) were found to be IC50 = 66.16 µg/mL and 76.05 µg/mL, respectively. E. sativa also showed promising anticancer activity against both the colorectal cancer cells HCT-116 (IC50 = 64.91 µg/mL) and Caco-2 (IC50 = 83.98 µg/mL) in a dose/time dependent manner. The phytoconstituents identified showed promising pharmacokinetics properties, representing a valuable source for drug or nutraceutical development. These investigations will lead to the further exploration as well as development of E. sativa-based nutraceutical products.