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

• Published in: Biotechnology letters 2012-02, Vol.34 (2), p.281-286
• Main Authors: Shinoda, Suguru, Kanamasa, Shin, Arai, Motoo
• Format: Article
• Language: English
• Subjects: Amino acids; Applied Microbiology; Bacillus circulans; Biochemistry; Biological and medical sciences; Biomedical and Life Sciences; Biotechnology; Cellulase; Cellulase - chemistry; Cellulase - genetics; Cellulase - metabolism; Cellulose; Cellulose - metabolism; Chitin; Chitosan; Chitosan - metabolism; Cloning; Cloning, Molecular; DNA, Bacterial - chemistry; DNA, Bacterial - genetics; Enzyme Stability; Enzymes; Fundamental and applied biological sciences. Psychology; Genes; Genetic recombination; Glucans - metabolism; Heating; Hydrogen-Ion Concentration; Life Sciences; Microbiology; Molecular Sequence Data; Molecular Weight; Original Research Paper; Paenibacillus; Paenibacillus - enzymology; Paenibacillus - genetics; Protein Sorting Signals; Recombinant; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Sequence Analysis, DNA; Sequence Homology, Amino Acid; Substrate Specificity; Temperature
• ISSN: 0141-5492; 1573-6776
• DOI: 10.1007/s10529-011-0759-5

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.