Cloning and characterization of a xylanase, KRICT PX1 from the strain Paenibacillus sp. HPL-001.

摘要: The KRICT PX1 gene (GB: FJ380951) consisting of 996bp encoding a protein 332 amino acids (38.1kDa) from the recently isolated Paenibacillus sp. strain HPL-001 (KCTC11365BP) has been cloned and expressed in Escherichia coli. xylanase showed high activity on birchwood xylan, was active over pH range 5.0 to 11.0, with two optima at 5.5 9.5 50 degrees C K(m) value 5.35 3.23, respectively. not affected by most salts, such as NaCl, LiCl, KCl, NH(4)Cl, CaCl(2), MgCl(2), MnCl(2), CsCl(2) 1mM, but CuSO(4), ZnSO(4), FeCl(3). One mM EDTA, 2-mercaptoethanol, PMSF did affect activity. TLC analysis catalyzed products after reaction xylan revealed that xylobiose major product smaller amounts xylotriose xylose. A similarity resulted 72% identity Geobacillus stearothermophilus ZP_03040360), 70% intracellular an uncultured bacterium AAP51133), 68% endo-1-4-xylanse ZP_02847150). In addition, acid alignment glycosyl hydralase (GH) family 10 xylanases degree homology highly conserved regions including catalytic sites, this confirmed through PROSITE scan. These results imply is new gene, alkaline belongs GH 10.