Pectinases are the group of enzymes that catalyze the degradation of pectic substances. It has wide applications in food industries for the production and clarification of wines and juices. The aim of this study was to isolate, screen and characterize pectinase from fungi isolated from various soil samples and evaluate its application in juice clarification. Fungal strains were isolated and screened primarily using 1% citruspectin incorporated potato dextrose agar (PDA) and secondarily using pectinase screening agar medium (PSAM) for pectinolytic organisms. The enzyme was produced by submerged state fermentation and assayed using the dinitro salicylic acid (DNS) method. From 20 different soil samples, 55 fungal isolates were screened primarily and, among them, only 14 isolates were subjected for secondary screening. Out of 14, only four strains showed the highest pectinolytic activity. Among four strains, Aspergillus spp. Gm showed the highest enzyme production at a 48-h incubation period, 1% substrate concentration, and 30 °C temperature. The thermal stability assessment resulted that the activity of pectinase enzyme declines by 50% within 10 min of heating at 60 °C. The optimum temperature, pH, and substrate concentration for the activity of enzyme was 30 °C (75.4 U/mL), 5.8 (72.3 U/mL), and 0.5% (112.0 U/mL), respectively. Furthermore, the yield of the orange juice, the total soluble solid (TSS), and clarity (% transmittance) was increased as the concentration of the pectinase increased, indicating its potential use in juice processing. Overall, the strain Aspergillus spp. Gm was identified as a potent strain for pectinase production in commercial scale.

Coffee pulp, mucilage, and beans with mucilage were used to develop alcoholic beverages. The pulp of 45.3% pulp, 54.7% mucilage with seed, and 9.4% mucilage only were obtained during the wet processing of coffee. Musts were prepared for all to TSS (Total soluble solid) 18 °Bx and fermentation was carried out for 12–16 days until TSS decreased to 5 °Bx at 30 °C. Phenolic characteristics, chromatic structures, chemical parameters, and sensory characteristics were analyzed for the prepared alcoholic beverages. Methanol content, ester content, aldehyde, alcohol, total acidity, caffeine, polyphenols, flavonoids, chromatic structure, and hue of the alcoholic beverage from the pulp was 335 mg/L, 70.58 ppm, 9.15 ppm, 8.86 ABV%, 0.41%, 30.94 ppm, 845.7 mg GAE/g dry extract, 440.7 mg QE/g dry extract, 0.41, and 1.71, respectively. An alcoholic beverage from the pulp was found superior to an alcoholic beverage from mucilage with beans and a beverage from mucilage in sensory analysis. There is the possibility of developing fermented alcoholic beverages from coffee pulp and mucilage. However, further research is necessary for quality of the beans that were obtained from the fermentation with the mucilage.

Moringa oleifera is a nutrient-rich plant, also referred to as a miracle tree, and is commonly used in the preparation of functional foods including herbal biscuits. Despite having a wide range of biomolecules, M. oleifera has not been studied for its nutritional benefits in Nepal. To fill this gap, five different formulations of flower and leaf powder ratios of 11:4, 11.75:3.25, 12.5:2.5, 13.25:1.75, 14:1 named as A, B, C, D, E, and control formulations were tested for their sensory and chemical characteristics. The results showed that calcium content (115.73 mg/100 g) was higher in biscuits with a higher percentage of the leaf (11:4) while TPC was minimum. Further, biscuits containing a higher percentage of flower powder contained fewer tannins. The sensory analysis concluded that D was deemed the best in overall attributes by panelists upon statistical analysis, however formulations A and B were superior to other samples regarding the chemical properties. These findings confirm that there is a huge potential for improving herbal biscuits.

Ginsenosides are the principal compounds responsible for the pharmacological effects and health benefits of Panax ginseng root. Among protopanaxadiol (PPD)-type ginsenosides, minor ginsenosides such as ginsenoside (G)-F2, G-Rh2, compound (C)-Mc1, C-Mc, C-O, C-Y, and C-K are known to be more pharmacologically active constituents than major ginsenosides such as G-Rb1, G-Rb2, G-Rc, and G-Rd. A novel ginsenoside Rc-hydrolyzing β-glucosidase (BG-1) from Armillaria mellea mycelia was purified as a single protein band with molecular weight of 121.5 kDa on SDS-PAGE and a specific activity of 17.9 U mg−1 protein. BG-1 concurrently hydrolyzed α-(1 → 6)-arabinofuranosidic linkage at the C-20 site or outer β-(1 → 2)-glucosidic linkage at the C-3 site of G-Rc to produce G-Rd and C-Mc1, respectively. The enzyme also hydrolyzed outer and inner glucosidic linkages at the C-3 site of G-Rd to produce C-K via G-F2, and inner glucosidic linkage at the C-3 site of C-Mc1 to produce C-Mc. C-Mc was also slowly hydrolyzed α-(1 → 6)-arabinofuranosidic linkage at the C-20 site to produce C-K with reaction time prolongation. Finally, the pathways for formation of C-Mc and C-K from G-Rc by BG-1 were G-Rc → C-Mc1 → C-Mc and G-Rc → G-Rd → G-F2 → C-K, respectively. The optimum reaction conditions for C-Mc and C-K formation from G-Rc by BG-1 were pH 4.0–4.5, temperature 45–60 °C, and reaction time 72–96 h. This is the first report of efficient production of minor ginsenosides, C-Mc and C-K from G-Rc by β-glucosidase purified from A. mellea mycelia.

Minor ginsenosides, such as compounds (C)-K and C-Y, possess relatively better bioactivity than those of naturally occurring major ginsenosides. Therefore, this study focused on the biotransformation of major ginsenosides into minor ginsenosides using crude β-glucosidase preparation isolated from submerged liquid culture of Fomitella fraxinea (FFEP). FFEP was prepared by ammonium sulfate (30–80%) precipitation from submerged culture of F. fraxinea. FFEP was used to prepare minor ginsenosides from protopanaxadiol (PPD)-type ginsenoside (PPDG-F) or total ginsenoside fraction (TG-F). In addition, biotransformation of major ginsenosides into minor ginsenosides as affected by reaction time and pH were investigated by TLC and HPLC analyses, and the metabolites were also identified by UPLC/negative-ESI-Q-TOF-MS analysis. FFEP biotransformed ginsenosides Rb1 and Rc into C-K via the following pathways: Rd → F2 → C-K for Rb1 and both Rd → F2→ C-K and C-Mc1 → C-Mc → C-K for Rc, respectively, while C-Y is formed from Rb2 via C-O. FFEP can be applied to produce minor ginsenosides C-K and C-Y from PPDG-F or TG-F. To the best of our knowledge, this study is the first to report the production of C-K and C-Y from major ginsenosides by basidiomycete F. fraxinea.

A total of 100 fresh-raw vegetable samples were collected from the Kathmandu, Lalitpur and Bhaktapur districts of Nepal to evaluate microbial and parasitic contamination, presence of Extended Spectrum Beta Lactamase (ESBL)-producing Escherichia coli and detect the bla_TEM and bla_CTX-M genes among the Escherichia coli isolates. This study revealed that the prevalence of Giardia cysts was highest (100%) and Hookworm and Entamoeba coli were lowest (1% each). Coliforms were isolated from every raw vegetable sample. A total of 178 bacterial isolates were isolated among which 57 isolates were identified as E. coli, out of which 33 were Multi-drug resistant (MDR) isolates. The high rate of resistance was found towards amoxicillin/clavulanate, tetracycline and cotrimoxazole. The 10 E. coli isolates tested positive in an ESBL screening, out of which 4 were confirmed as ESBL producers by a combined disc test. Out of these 4 confirmed ESBL E. coli, one was found to carry both the bla_TEM gene and bla_CTX-M genes by the Polymerase Chain Reaction (PCR) technique. One isolate has only the bla_TEM gene, while other isolate harboured only bla_CTX-M genes.