Xylooligosaccharides production by crude microbial enzymes from agricultural waste without prior treatment and their potential application as nutraceuticals.

Aspergillus fumigatus R1, on submerged fermentation using agricultural residues as carbon source produced extracellular xylanase (152IU/ml after 96h of incubation at 37°C with constant shaking at 100rpm). A maximum yield of 1gm% Xylooligosaccharides (XOS) mixture was obtained after 12h by enzymatic hydrolysis of xylan rich wheat husk without any prior pretreatment using the crude enzyme without any purification. HP-TLC data confirmed the presence of an array of XOS for its prebiotic properties by carrying out studies on ten standard probiotic cultures. Six of ten probiotic cultures were able to utilize XOS produced from agricultural wastes and showed remarkable growth in the media containing XOS as the sole source of carbon. XOS mixture also exhibited concentration dependent anti-oxidant activity. Thus, the results showed that XOS produced from agricultural residues have great prebiotic potential and good antioxidant activity; therefore, it can be used in food-related applications.

Purification, biochemical characterization and structural modelling of alkali-stable ß-1,4-xylan xylanohydrolase from Aspergillus fumigatus R1 isolated from soil.

BACKGROUND: Aspergillus fumigatus R1 produced xylanase under submerged fermentation which degrades the complex hemicelluloses contained in agricultural substrates. Xylanases have gained considerable attention because of their tremendous applications in industries. The purpose of our study was to purify xylanase and study its biochemical properties. We have predicted the secondary structure of purified xylanase and evaluated its active site residues and substrate binding sites based on the global and local structural similarity. RESULTS: Various microorganisms were isolated from Puducherry soil and screened by Congo-red test. The best isolate was identified to be Aspergillus fumigatus R1. The production kinetics showed the highest xylanase production (208 IU/ml) by this organism in 96 h using 1 % rice bran as the only carbon source. The purification of extracellular xylanase was carried out by fractional ammonium sulphate precipitation (30-55 %), followed by extensive dialysis and Bio-Gel P-60 Gel-filtration chromatography. The enzyme was purified 58.10 folds with a specific activity of 38196.22 IU/mg. The biochemical characterization of the pure enzyme was carried out for its optimum pH and temperature (5.0 and 50(0)C), pH and temperature stability, molecular mass (Mr) (24.5 kDa) and pI (6.29). The complete sequence of protein was obtained by mass spectrometry analysis. Apparent Km and Vmax values of the xylanase for birchwood xylan were 11.66 mg/ml and 87.6 µmol min(-1) mg(-1) respectively. CONCLUSION: Purified xylanase was analyzed by mass-spectrometry which revealed 2 unique peptides. Xylanase under current study showed significant production using agricultural residues and a broad range of pH stability in the alkaline region. Xylanase produced by Aspergillus fumigatus R1 could serve as the enzyme of choice in industries.