The aim of the present study was to produce exo-polygalacturonase from potent soil isolate by submerged fermentation and its application for fruit juice treatment. Pectinase producing strains were selectively isolated from pectin industry waste. A selected isolate C2 was found to produce significant amount of exo-polygalacturonase. The isolate was identified as Paecilomyces variotii on the basis of morphological characteristics and 18S rRNA gene sequence analysis. The exo-polygalacturonase produced by the isolate was purified by ammonium sulphate precipitation, size exclusion chromatography and ion exchange chromatography. The purified enzyme had MW of 39.4 kD based on SDS PAGE. Under partially optimized conditions, purified exo-polygalacturonase showed specific activity of 98.49 U/mg protein at pH 6.0 and 30°C. The enzyme was comparatively stable from 10 to 30°C and the activity decreased with increasing temperature. Purified enzyme brought about considerable reduction in viscosity of fruit juice samples.
Bacillus spp. ST13, isolated from human stool, was evaluated for siderophoregenic and probiotic qualities prior to its possible application for iron nutrition in humans and animals. It was tested for siderophore production in iron-limiting conditions and found to produce catecholate type of siderophore on the basis of high-performance liquid chromatography (HPLC), FT-IR, NMR, and mass spectra analysis. The isolate was screened for probiotic properties as per WHO and FAO guidelines. The strain ST13 can survive stomach acidity, bile salt and partially simulated gastrointestinal tract conditions. It was susceptible to most of the antibiotic tested and showed antimicrobial activity against enteric pathogens like Salmonella typhimurium, Streptococcus pyogenes, and Staphylococcus aureus. Strain ST13 showed close similarity with Bacillus subtilis using 16S r-RNA gene sequence analysis and biochemical characterization. The methanolic extract of ST13 siderophore was evaluated for DPPH radical scavenging activity, which showed 94.55 ± 0.9% of radical scavenging effect.
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