Comparative assessment of solvents and lignocellulolytic enzymes affiliated extraction of polyphenols from the various lignocellulosic agro-residues: identification and their antioxidant properties.

The present investigation was aimed to utilize lignocellulosic agro-residues and compare the extraction of polyphenols utilizing lignocellulolytic enzymes secreted by Sphingobacterium sp. ksn and with that of the solvents (ethanol, methanol) affiliated methods. The maximum amount of polyphenols, flavonoids and tannins were 94.29, 11.36, and 79.21 g 100 g-1 respectively, found in the extracts obtained by enzymes affiliated extraction of coffee cherry husk (CCH). The phenolics namely, gallic acid, caffeic acid, coumaric acid, 1-hydroxybenzoic acid, 2,5-dihydroxybenzoic acid, p-hydroxybenzaldehyde were commonly found whereas syringic acid, quercetin, kaempferol, and epicatechin were hardly found in the extracts of agro-residues. The extracts of CCH shown maximum antioxidant properties for DPPH, ABTS, and FRAP. The present study reports that the affiliation of enzymes for the extraction of polyphenols from agro-residues is more efficient than that of the solvents affiliation and CCH as the good source of polyphenols.

Isolation and Characterization of Gut Bacterial Proteases Involved in Inducing Pathogenicity of Bacillus thuringiensis Toxin in Cotton Bollworm, Helicoverpa armigera.

Bacillus thuringiensis toxin proteins are deployed in transgenic plants for pest management. The present studies were aimed at characterization of gut bacterial proteases involved in activation of inactive Cry1Ac protoxin (pro-Cry1Ac) to active toxin in Helicoverpa armigera. Bacterial strains were isolated from H. armigera midgut and screened for their proteolytic activation toward pro-Cry1Ac. Among 12 gut bacterial isolates seven isolates showed proteolytic activity, and proteases from three isolates (IVS1, IVS2, and IVS3) were found to be involved in the proteolytic conversion of pro-Cry1Ac into active toxin. The proteases from IVS1, IVS2, and IVS3 isolates were purified to 11.90-, 15.50-, and 17.20-fold, respectively. The optimum pH and temperature for gut bacterial protease activity was 8.0 and 40°C. Maximum inhibition of total proteolytic activity was exerted by phenylmethane sulfonyl fluoride followed by EDTA. Fluorescence zymography revealed that proteases from IVS1, IVS2, and IVS3 were chymotrypsin-like and showing protease band at ~15, 65, and 15 kDa, respectively. Active Cry1Ac formed from processing pro-Cry1Ac by gut bacterial proteases exhibited toxicity toward H. armigera. The gut bacterial isolates IVS1, IVS2, and IVS3 showed homology with B. thuringiensis (CP003763.1), Vibrio fischeri (CP000020.2), and Escherichia coli (CP011342.1), respectively. Proteases produced by midgut bacteria are involved in proteolytic processing of B. thuringiensis protoxin and play a major role in inducing pathogenicity of B. thuringiensis toxins in H. armigera.