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Molecular Cloning and Gene Expression Analysis of Tomato Endo-β- N -acetylglucosaminidase, an Endoglycosidase Involved in the Production of High-Mannose Type Free N -Glycans during Tomato Fruit Ripening

2009; Oxford University Press; Volume: 73; Issue: 2 Linguagem: Inglês

10.1271/bbb.80889

1347-6947

Kôsuke Nakamura, Masami Inoue, Megumi Maeda, Ryohei Nakano, Katsutoshi Hosoi, Kazuhito Fujiyama, Yoshinobu Kimura,

Enzyme Production and Characterization

In this study, we identified a gene encoding tomato ENGase (Endo-LE) using the gene information of rice ENGase, and expressed the Endo-LE protein in Escherichia coli. The substrate specificity of the recombinant Endo-LE was the same as that of the native enzyme, showing strong activity towards the high-mannose type N-glycans with the Manalpha1-2Manalpha1-3Manbeta1-4GlcNAcbeta1-4GlcNAc unit. Real-time PCR analysis revealed that the gene expression of Endo-LE did not vary significantly with the tomato ripening process, indicating that Endo-LE activity is ubiquitously expressed.