Molecular cloning and gene expression analysis of tomato endo-beta-N-acetylglucosaminidase, an endoglycosidase involved in the production of high-mannose type free N-glycans during tomato fruit ripening.
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.
Nakamura
Kosuke
K
Department of Biofunctional Chemistry, Graduate School of Natural Science and Technology, Okayama University, Japan.
Inoue
Masami
M
Maeda
Megumi
M
Nakano
Ryohei
R
Hosoi
Katsutoshi
K
Fujiyama
Kazuhito
K
Kimura
Yoshinobu
Y
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D005638:Fruit / Q000235:genetics / Q000254:growth & development*; D018506:Gene Expression Regulation, Plant; D018551:Solanum lycopersicum / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development*; D008358:Mannose / Q000378:metabolism*; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D011134:Polysaccharides / Q000096:biosynthesis* / Q000737:chemistry; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D013379:Substrate Specificity
D011134:Polysaccharides; D012333:RNA, Messenger; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D008358:Mannose