Characterization of an Extracellular Xylanase in Paenibacillus sp. HY-8 Isolated from an Herbivorous Longicorn Beetle

• Published in: Journal of microbiology and biotechnology Vol. 16; no. 11; pp. 1753 - 1759
• Main Authors: Heo, Sun-Yeon, Kwak, Jang-Yul, Oh, Hyun-Woo, Park, Doo-Sang, Bae, Kyung-Sook, Shin, Dong-Ha, Park, Ho-Yong
• Format: Journal Article
• Language: Korean
• Published: 한국미생물생명공학회 30.11.2006
• Subjects: Paenibacillus; purification; xylan; xylanase; xylan; Paenibacillus; purification; xylanase
• ISSN: 1017-7825; 1738-8872

Paenibacillus sp. HY-8 isolated from the digestive tracts of the longicorn beetle, Moechotypa diphysis, produced an extracellular endoxylanase with a molecular weight of 20 kDa estimated by SDS-PAGE. The xylanase was purified to near electrophoretic homogeneity from the culture supernatant after ammonium sulfate precipitation, gel filtration, and ionexchange chromatography. The purified xylanase exhibited the highest activities at pH 6.0 and $50^{\circ}C$. The $K_m\;and\;V_{max}$ values were 7.2 mg/ml and 16.3 U/mg, respectively, for birchwood xylan as the substrate. Nucleotide sequence of the PCR-cloned gene was determined to have the open reading frame encoding a polypeptide of 212 amino acids. The N-terminal amino acid sequence and the nucleotide sequence analyses predicted that the precursor xylanase contained a signal peptide composed of 28 amino acids and a catalytically active 19.9-kDa peptide fragment. The deduced amino acid sequence shared extensive similarity with those of the glycoside hydrolase family 11 of xylanases from other bacteria. The predicted amino acid sequence contained two glutamate residues, previously identified as essential and conserved for active sites in other xylanases of the glycoside hydrolase family 11.