Characterization of a xylanase from a thermophilic strain of Anoxybacillus pushchinoensis A8

• Published in: Biológia 2008-10, Vol.63 (5), p.599-606
• Main Authors: Kacagan, Murat, Sabriye Canakci, Cemal Sandalli, Kadriye Inan, Dilsat Colak, Ali Belduz
• Format: Article
• Language: English
• Subjects: Anoxybacillus; Anoxybacillus pushchinoensis; Biomedical and Life Sciences; Cell Biology; chromatography; DNA; exoxylanase; hot springs; Life Sciences; Microbiology; moderately thermophilic; molecular weight; nucleic acid hybridization; nucleotide sequences; oats; Plant Sciences; ribosomal RNA; temperature; thermostable xylanase; xylan; xylanases; xylanolytic microorganisms; xylooligosaccharides; xylose; Zoology
• ISSN: 1336-9563; 0006-3088; 1336-9563
• DOI: 10.2478/s11756-008-0134-8

A facultatively anaerobic, thermophilic, xylanolytic bacterium was isolated from a sample collected from the Diyadin Hot Springs, Turkey. According to morphological, biochemical and molecular identification, this new strain was suggested to be representative of the Anoxybacillus pushchinoensis and it was designated as Anoxybacillus pushchinoensis strain A8. It exhibited 97% similarity to 16S rRNA gene sequence of A. pushchinoensis and 77% DNA homology by DNA-DNA hybridization studies. Q-sepharose and CM-sepharose chromatography was used to purify an extracellular xylanase to >90% purity from this species. The enzyme had a molecular mass of approximately 83 kDa. The enzyme showed optimum activity at pH 6.5 and it was 96% stable over a broad pH range of 6.5–11 for 24 hours. The enzyme had optimum activity at 55°C and it was 100% stable at temperature between 50–60°C up to 24 hours. Kinetic characterization of the enzyme was performed at temperature optima (55°C) and Vmax and K m were found to be 59.88 U/mg protein and 0.909 mg/mL, respectively. Oat spelt xylan but not xylooligosaccharides was degraded by the enzyme and xylose was the only product detected from oat xylan degradation. This suggested that the enzyme was an exo-acting xylanase.