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

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

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

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 highmannose type N-glycans with the Manβ1-2Manβ1- 3Manβ1- 4GlcNAcβ1-4GlcNAc 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.

Original languageEnglish
Pages (from-to)461-464
Number of pages4
JournalBioscience, Biotechnology and Biochemistry
Volume73
Issue number2
DOIs
Publication statusPublished - 2009

Fingerprint

Acetylglucosaminidase
Gene encoding
Cloning
Glycoside Hydrolases
Molecular Cloning
Lycopersicon esculentum
Mannose
Fruits
Gene expression
Escherichia coli
Polysaccharides
Fruit
Genes
Gene Expression
Escherichia coli Proteins
Substrates
Enzymes
Substrate Specificity
Real-Time Polymerase Chain Reaction
Proteins

Keywords

  • De-N-glycosylation
  • Endo-β-N-acetylglucosaminidase
  • Free Nglycans
  • Lycopersicon esculentu
  • Tomato fruit ripening

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Applied Microbiology and Biotechnology
  • Analytical Chemistry
  • Organic Chemistry
  • Medicine(all)

Cite this

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title = "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",
abstract = "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 highmannose type N-glycans with the Manβ1-2Manβ1- 3Manβ1- 4GlcNAcβ1-4GlcNAc 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.",
keywords = "De-N-glycosylation, Endo-β-N-acetylglucosaminidase, Free Nglycans, Lycopersicon esculentu, Tomato fruit ripening",
author = "Kosuke Nakamura and Masami Inoue and Megumi Maeda and Ryohei Nakano and Katsutoshi Hosoi and Kazuhito Fujiyama and Yoshinobu Kimura",
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T1 - 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

AU - Nakamura, Kosuke

AU - Inoue, Masami

AU - Maeda, Megumi

AU - Nakano, Ryohei

AU - Hosoi, Katsutoshi

AU - Fujiyama, Kazuhito

AU - Kimura, Yoshinobu

PY - 2009

Y1 - 2009

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AB - 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 highmannose type N-glycans with the Manβ1-2Manβ1- 3Manβ1- 4GlcNAcβ1-4GlcNAc 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.

KW - De-N-glycosylation

KW - Endo-β-N-acetylglucosaminidase

KW - Free Nglycans

KW - Lycopersicon esculentu

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