title: Purification, biochemical characterization and structural modelling of alkali-stable β-1,4-xylan xylanohydrolase from Aspergillus fumigatus R1 isolated from soil.
abstract: 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.
  
  
  CONCLUSIONS
  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.
authors: Department of Microbiology, School Of Life-Sciences, Pondicherry University, R.V. Nagar, Kalapet, Puducherry, 605014, India. rrehandeshmukh@gmail.com.
authors: Rehan Ahmed
authors: RA
authors: Deshmukh
authors: Department of Microbiology, School Of Life-Sciences, Pondicherry University, R.V. Nagar, Kalapet, Puducherry, 605014, India. sharmili.pumicro@gmail.com.
authors: Sharmili
authors: S
authors: Jagtap
authors: Department of Microbiology, School Of Life-Sciences, Pondicherry University, R.V. Nagar, Kalapet, Puducherry, 605014, India. madan07bsmmt@gmail.com.
authors: Madan Kumar
authors: MK
authors: Mandal
authors: Department of Microbiology, School Of Life-Sciences, Pondicherry University, R.V. Nagar, Kalapet, Puducherry, 605014, India. smcoolsuraj@gmail.com.
authors: Suraj Kumar
authors: SK
authors: Mandal
mesh_terms: D000595:Amino Acid Sequence; D001232:Aspergillus fumigatus / Q000201:enzymology* / Q000235:genetics; D043364:Endo-1,4-beta Xylanases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D012988:Soil Microbiology; D013696:Temperature
chemical_list: D043364:Endo-1,4-beta Xylanases