Purification and molecular cloning of xylanases from the wood-feeding termite, Coptotermes formosanus Shiraki

• Published in: Bioscience, biotechnology, and biochemistry Vol. 73; no. 3; pp. 710 - 718
• Main Authors: Arakawa, G.(Ryukyu Univ., Nishihara, Okinawa (Japan)), Watanabe, H, Yamasaki, H, Maekawa, H, Tokuda, G
• Format: Journal Article
• Language: English
• Published: Tokyo Japan Society for Bioscience, Biotechnology, and Agrochemistry 23.03.2009; Japan Society for Bioscience Biotechnology and Agrochemistry; Oxford University Press
• Subjects: Amino Acid Sequence; Animals; APPAREIL DIGESTIF; Base Sequence; Biological and medical sciences; Chromatography; CLONACION MOLECULAR; CLONAGE MOLECULAIRE; Cloning, Molecular; Coptotermes formosanus; DEGRADACION; DEGRADATION; DIGESTIVE SYSTEM; DNA, Complementary - genetics; ENZIMAS; ENZYME; ENZYMES; Fundamental and applied biological sciences. Psychology; Gastrointestinal Tract - enzymology; GHF11; ISOPTERA; Isoptera - enzymology; Isoptera - genetics; Isoptera - physiology; MASTIGOPHORA; MOLECULAR CLONING; Molecular Sequence Data; Nucleic Acid Amplification Techniques; PCR; protist; PURIFICACION; PURIFICATION; SIMBIONTICO; SISTEMA DIGESTIVO; SYMBIONTS; Symbiosis; SYMBIOTE; termite; XILANOS; xylan; xylanase; XYLANE; XYLANS; Xylosidases - chemistry; Xylosidases - genetics; Xylosidases - isolation & purification; Xylosidases - metabolism; Purification; Wood; Enzyme; Insecta; Coptotermes formosanus; Isoptera; Xylan; Endo-1,4-β-xylanase; GHF11; xylanase; Glycosylases; Termite; Arthropoda; Glycosidases; protist; Hydrolases; Rhinotermitidae; Xylan endo-1,3-β-xylosidase; Invertebrata; Molecular cloning
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1271/bbb.80788

Coptotermes formosanus is one of the most destructive termites in the southern part of Japan as well as in the United States. Hemicellulose is a noncellulosic polysaccharide found in plant cell walls, and xylan is the major constituent of hemicellulose. Since hemicellulose prevents access of cellulolytic enzymes to cellulose, enzymatic hydrolysis of hemicellulose is beneficial for cellulose digestion. We purified three functional xylanases to homogeneity from C. formosanus for the first time. Elution profiles from the whole termite extract suggest that these three xylanases play major roles in xylan digestion in the gut of the termites. The corresponding cDNAs were successfully cloned based on the N-terminal amino acid sequences, encoding GHF11 xylanases. Reverse transcription-PCR using manipulated protozoan cells in the hindgut revealed that the corresponding genes were expressed in the symbiotic flagellate Holomastigotoides mirabile. These results suggest that the GHF11 xylanases that are produced by the symbiotic flagellates play a primary role in xylan degradation in C. formosanus.