Biotechnology Letters

, Volume 34, Issue 2, pp 281–286 | Cite as

Cloning of an endoglycanase gene from Paenibacillus cookii and characterization of the recombinant enzyme

  • Suguru Shinoda
  • Shin Kanamasa
  • Motoo Arai
Original Research Paper


An endoglycanase gene of Paenibacillus cookii SS-24 was cloned and sequenced. This Pgl8A gene had an open reading frame of 1,230 bp that encoded a putative signal sequence (31 amino acids) and mature enzyme (378 amino acids: 41,835 Da). The enzyme was most homologous to a β-1,3-1,4-glucanase of Bacillus circulans WL-12 with 84% identity. The recombinant enzyme hydrolyzed carboxymethyl cellulose, swollen celluloses, chitosan and lichenan but not Avicel, chitin powder or xylan. With chitosan as the substrate, the optimum temperature and hydrolysis products of the recombinant enzyme varied at pH 4.0 and 8.0. This is the first report that characterizes chitosanase activity under different pH conditions.


Cellulase Chitosanase Gene cloning Paenibacillus cookii 

Supplementary material

10529_2011_759_MOESM1_ESM.docx (14 kb)
Supplementary Table 1. (DOCX 15 kb)
10529_2011_759_MOESM2_ESM.docx (93 kb)
Supplementary Fig. 1. SDS–PAGE analysis of the purified enzyme from P. cookii SS-24 and recombinant enzyme. Lane M, protein standard marker (Protein Ladder 10-250 kDa, New England Biolabs); Lane 1, purified enzyme from P. cookii; Lane 2, purified recombinant enzyme. The purified enzymes were applied to 12% SDS–PAGE. (DOCX 94 kb)
10529_2011_759_MOESM3_ESM.docx (1.3 mb)
Supplementary Fig. 2. Comparison of the amino acid sequence of Pgl8A and other glycosyl hydrolases. The amino acid sequences of GHF-8 enzymes from P. cookii SS-24 (Pgl8A), B. circulans WL-12 (B.WL-12), B. circulans KSM-N257 (B.N257), Lysobacter sp. IB-9374 (L.IB9374), Bacillus sp. KCTC0377BP (B. KCTC) and Bacillus sp. K17 (B.K17) are shown. The conserved amino acid sequence of GHF-8 catalytic module is underlined. Asterisks denote potential catalytic residues. (DOCX 1,305 kb)


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Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  1. 1.Department of Environmental Biology, College of Bioscience and BiotechnologyChubu UniversityAichiJapan

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