グライコシンターゼ化による反転型加水分解酵素のグリコシド合成触媒への変換
スポンサーリンク
概要
- 論文の詳細を見る
還元末端キシロース遊離エキソオリゴキシラナーゼ（Rex, EC. 3.2.1.156）は，糖加水分解酵素ファミリー 8 に属する反転型キシラン分解酵素であり，キシロオリゴ糖から還元末端キシロース（X1）を遊離する反応を触媒する。Rex は X1 の存在下のみ，α キシロビオシルフルオリド（α-X2F）を加水分解する，Hehre-合成-加水分解反応を示した。一般塩基触媒残基（D263）変異酵素ライブラリーを作製し，グライコシンターゼ活性により α-X2F と X1 からキシロトリオース（X3）を最も良く蓄積する D263C 変異酵素を選抜した。しかしながら，D263C のフッ素イオン遊離活性は親酵素より低かった。次に，一般塩基触媒残基と共に求核試薬である水を保持している残基である Y198 をフェニルアラニンに変異させたところ，フッ素イオン遊離活性が低下することなく加水分解活性のみに大幅に低下が見られた。Rex の Y198F 変異酵素は D263C よりも X3 の蓄積が多かった。最近になり，糖加水分解酵素ファミリー 95 に属する α1,2 フコシダーゼのグライコシンターゼ化が報告された。この場合は，一般塩基触媒残基と相互作用している別の残基に変異を入れることが有効であった。両者ともに，一般塩基触媒残基は変異させない方が良かった。
- FCCA(Forum: Carbohydrates Coming of Age)の論文
著者
-
Kitaoka Motomitsu
National Food Research Institute, National Agriculture and Food Research Organization
-
Fushinobu Shinya
Department of Biotechnology, The University of Tokyo
-
Fushinobu Shinya
Dep. Of Biotechnology Graduate School Of Agricultural And Life Sciences The Univ. Of Tokyo
-
Kitaoka Motomitsu
Enzyme Laboratory National Food Research Institute National Agriculture And Food Research Organizati
-
Honda Yuji
National Food Research Institute, National Agriculture and Food Research Organization
-
Hidaka Masafumi
National Food Research Institute, National Agriculture and Food Research Organization
-
Katayama Takane
Ishikawa Prefectural University
-
Yamamoto Kenji
Division of Integrated Life Science, Graduate School of Biostudies, Kyoto University
関連論文
- Practical Preparation of D-Galactosyl-β1→4-L-rhamnose Employing the Combined Action of Phosphorylases
- A Eukaryotic Copper-Containing Nitrite Reductase Derived from a NirK Homolog Gene of Aspergillus oryzae
- Analyses of Bifidobacterial Glycosidases Involved in the Metabolism of Oligosaccharides
- The Role of Conserved Arginine Residue in Loop 4 of Glycoside Hydrolase Family 10 Xylanases
- Crystallographic and Mutational Analyses of Substrate Recognition of Endo-α-N-acetylgalactosaminidase from Bifidobacterium longum
- A Series of Crystal Structures of a meta-Cleavage Product Hydrolase from Pseudomonas fluorescens IP01 (CumD) Complexed with Various Cleavage Products
- Purification, Characterization, and Steady-State Kinetics of a meta-Cleavage Compound Hydrolase from Pseudomonas fluorescens IP01
- The Possible Involvement of Copper-Containing Nitrite Reductase (NirK) and Flavohemoglobin in Denitrification by the Fungus Cylindrocarpon tonkinense
- Streptomyces griseus Enhances Denitrification by Ralstonia pickettii K50, Which Is Possibly Mediated by Histidine Produced during Co-Culture
- グライコシンターゼ化による反転型加水分解酵素のグリコシド合成触媒への変換
- N-Linked Oligosaccharides of Aspergillus awamori Feruloyl Esterase Are Important for Thermostability and Catalysis
- Involvement of a Glu71-Arg64 Couple in the Access Channel for NADH in Cytochrome P450nor
- Kinetic Studies on p-Nitrophenyl-cellobioside Hydrolyzing Xylanase from Cellvibrio gilvus
- Phosphorolytic Reaction of Cellvibrio gilvus Cellobiose Phosphorylase
- Purification and Properties of a Xylanase from Cellvibrio gilvus That Hydrolyzes p-Nitrophenyl Cellooligosaccharides(Biological Chemistry)
- Cloning of a Gene Encoding a Highly Stable Endo-β-1, 4-Glucanase from Aspergillus niger and Its Expression in Yeast
- Transcriptional Regulation of Tyrosine Phenol-lyase Gene Mediated through TyrR and cAMP Receptor Protein
- Transglycosylation Activity of the Endo-β-1, 4-Glucanase from Aspergillus niger IFO31125 and Its Application
- Effects of Carbohydrate Chain on Surface Net Charge and Hydrophobicity of Glycoenzymes
- Purification and Characterization of Thermostable α-Galactosidase from Ganoderma lucidum(Biochemistry & Molecular Biology)
- 647 A putative proline iminopeptidase of Thermotoga maritime with leucine- and lysine-p-nitroanilide hydrolyzing activities
- Practical Preparation of Lacto-N-biose I, a Candidate for the Bifidus Factor in Human Milk
- Characterization of a Cellobiose Phosphorylase from a Hyperthermophilic Eubacterium, Thermotoga maritima MSB8(Biochemistry & Molecular Biology)
- Kinetic Studies of a Recombinant Cellobiose Phosphorylase (CBP) of the Clostridium thermocellum YM4 Strain Expressed in Escherichia coli
- Effects of Truncation at the Non-homologous Region of a Family 3 β-Glucosidase from Agrobacterium tumefaciens
- Identification of the Putative Proton Donor Residue of Lacto-N-biose Phosphorylase (EC 2.4.1.211)
- Syntheses of 4-Methylumbelliferyl-β-D-Xylobioside and 5-Bromo-3-Indolyl-β-D-Xylobioside for Sensitive Detection of Xylanase Activity on Agar Plates
- Diversity and Similarity of Microbial Communities in Petroleum Crude Oils Produced in Asia
- Characterization of Endo-β-N-acetylglucosaminidase from Alkaliphilic Bacillus halodurans C-125
- Plate Assay for Endo-β-N-Acetylglucosaminidase Activity Using a Chromogenic Substrate Synthesized by Transglycosylation with Arthrobacter Endo-β-N-Acetylglucosaminidase
- Unique Peptide:N-glycanase of Caenorhabditis elegans has Activity of Protein Disulphide Reductase as well as of Deglycosylation
- Unique Sugar Metabolic Pathways of Bifidobacteria
- Characterization of Bacillus halodurans α-Galactosidase Mel4A Encoded by the mel4A Gene (BH2228)
- Characterization of a Thermostable Family 10 Endo-Xylanase (XynB) from Thermotoga maritima That Cleaves p-Nitrophenyl-β-D-Xlyloside
- Analyses of Bifidobacterial Glycosidases Involved in the Metabolism of Oligosaccharides
- Kinetic Studies on the Hydrolysis of N-Acetylated and N-Deacetylated Derivatives of 4-Methylumbelliferyl Chitobioside by the Family 18 Chitinases ChiA and ChiB from Serratia Marcescens
- Substrate Specificity of the N, 6-ο-Diacetylmuramidase from Streptomyces globisporus
- グライコシンターゼ化による反転型加水分解酵素のグリコシド合成触媒への変換
- A Eukaryotic Copper-Containing Nitrite Reductase Derived from a NirK Homolog Gene of Aspergillus oryzae
- Structural explanation for the acquisition of glycosynthase activity
- Prebiotic Effect of Lacto-N-biose I on Bifidobacterial Growth
- The Possible Involvement of Copper-Containing Nitrite Reductase (NirK) and Flavohemoglobin in Denitrification by the Fungus Cylindrocarpon tonkinense
- Streptomyces griseus Enhances Denitrification by Ralstonia pickettii K50, Which Is Possibly Mediated by Histidine Produced during Co-Culture
- Characterization of Bacillus halodurans α-Galactosidase Me14A Encoded by the mel4A Gene (BH2228)
- Practical Preparation of D-Galactosyl-β1→4-L-rhamnose Employing the Combined Action of Phosphorylases
- Molecular Anatomy of the Alkaliphilic Xylanase from Bacillus halodurans C-125
- Employing Chimeric Xylanases to Identify Regions of an Alkaline Xylanase Participating in Fnzyme Activity at Basic pH(ENZYMOLOGY, PROTEIN ENGINEERING, AND ENZYME TECHNOLOGY)
- Unique Sugar Metabolic Pathways of Bifidobacteria
- Structural Explanation for the Acquisition of Glycosynthase Activity
- Mutational Analysis of Fungal Family 11 Xylanases on pH Optimum Determination
- Interactions between Glycoside Hydrolase Family 94 Cellobiose Phosphorylase and Glucosidase Inhibitors
- Self-transferring Product Inhibition Observed during the Hydrolysis of Aryl-β-D-Glucopyranosides by a β-Glucosidase from Agrobacterium tumefaciens
- Characterization of a Bacterial Laminaribiose Phosphorylase
- Characterization of the Cytosolic β-N-Acetylglucosaminidase from Bifidobacterium longum subsp. longum