Molecular cloning and characterization of a glycosyl hydrolase family 9 cellulase distributed throughout the digestive tract of the cricket Teleogryllus emma |
| |
Authors: | Kim Namjung Choo Young Moo Lee Kwang Sik Hong Seong Jin Seol Kwang Youl Je Yeon Ho Sohn Hung Dae Jin Byung Rae |
| |
Institution: | aCollege of Natural Resources and Life Science, Dong-A University, Busan 604-714, South Korea;bDepartment of Agricultural Biology, National Institute of Agricultural Science and Technology, Suwon 441-100, South Korea;cSchool of Agricultural Biotechnology, Seoul National University, Seoul, South Korea |
| |
Abstract: | A novel endogenous β-1,4-endoglucanase (EG) gene belonging to the glycosyl hydrolase family 9 (GHF 9) that is distributed throughout the digestive tract of the cricket Teleogryllus emma was cloned and characterized. This gene, named TeEG-I, consists of eight exons encoding 453 amino acid residues and exists as a single copy in the T. emma genome. TeEG-I possesses all the features, including signature motifs and catalytic domains, of GHF 9 members, sharing high levels of identity with the termite, Mastotermes darwiniensis (64% protein sequence identity), and the cockroach, Panesthia cribrata (62%), GHF 9 cellulases. Recombinant TeEG-I, which is expressed as a 47-kDa polypeptide in baculovirus-infected insect Sf9 cells, showed an optimal pH and temperature of pH 5.0 and 40 °C. The Km and Vmax values for digestion of carboxymethyl cellulose were 5.4 mg/ml and 3118.4 U/mg, respectively. Northern and Western blot analyses revealed that TeEG-I is present throughout the digestive tract, which correlated with the TeEG-I distribution and cellulase activity in the digestive tract as assayed by immunofluorescence staining and enzyme activity assay, respectively. These results indicate that TeEG-I is distributed throughout the entire digestive tract of T. emma, suggesting a functional role of endogenous TeEG-I in a sequential cellulose digestion process throughout the T. emma digestion tract. |
| |
Keywords: | Cricket Teleogryllus emma Cellulase Endoglucanase Enzyme Glycoside hydrolase family 9 (GHF 9) Insect |
本文献已被 ScienceDirect PubMed 等数据库收录! |
|