We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
Xylanase from a soil isolate, Bacillus pumilus: gene isolation, enzyme production, purification, characterization and one-step separation by aqueous-two-phase system.
- Authors
Ersayin Yasinok, Aysegul; Biran, Suzan; Kocabas, Aytac; Bakir, Ufuk
- Abstract
A xylanase producer, Bacillus pumilus SB-M13, was isolated from soil and identified using various tests based on carbohydrate fermentation preferences and fatty acid analysis. Xylanase gene, isolated using PCR amplification, was partially sequenced and it showed 89–94% sequence similarity to the xylanase genes of other B. pumilus strains. Xylanase with very low level of cellulase was produced on agricultural byproducts. The enzyme has been purified 186-fold by hydrophobic interaction chromatography and biochemically characterized. It has a molecular weight of 24.8 kDa and pI of 9.2. Xylanolytic activity is stable at alkaline pH and highest activity is observed at 60 °C and pH 7.5. Enzyme Km and kcat values were determined as 1.9 mg/mL and 42,600 U/mg, respectively. In aqueous-two-phase system, xylanase always partitioned to the top phase. Basic pH, low PEG concentration, salt addition, and presence of microbial cells enhanced xylanase partitioning. A maximum sevenfold purification, 10-fold concentration and 100% xylanase recovery were obtained, separately, by adjusting system parameters. A fourfold concentrated xylanase was obtained with 70% enzyme recovery only in one step ATPS process without cell harvesting.
- Subjects
XYLANASES; BACILLUS (Bacteria); PROTEIN fractionation; CARBOHYDRATES; POLYMERASE chain reaction; CELLULASE; AGRICULTURAL wastes; MICROBIAL cell cycle
- Publication
World Journal of Microbiology & Biotechnology, 2010, Vol 26, Issue 9, p1641
- ISSN
0959-3993
- Publication type
Article
- DOI
10.1007/s11274-010-0340-8