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Literature summary extracted from

  • Ueno, K.; Sonoda, T.; Yoshida, M.; Shiomi, N.; Onodera, S.
    Purification, characterization, and functional analysis of a novel 6G&1-FEH mainly hydrolyzing neokestose from Asparagus (2018), J. Exp. Bot., 69, 4295-4308 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.80 gene aoeh4a, DNA and amino acid sequence determination and analysis Asparagus officinalis
3.2.1.80 gene aoeh4b, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis, recombinant expression in Pichia pastoris Asparagus officinalis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.80 130000
-
 native enzyme, gel filtration Asparagus officinalis

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.80 Asparagus officinalis A0A2Z6G7P5 aoeh4b; cvs. Gijnlim and Taihouwase
-
3.2.1.80 Asparagus officinalis A0A2Z6G7T9 aoeh4a; cv. Gijnlim and Taihouwase
-

Posttranslational Modification

EC Number Posttranslational Modification Comment Organism
3.2.1.80 glycoprotein the amino acid sequence contains four putative N-glycosylation sites (N-X-S/T). Deglycosylation of recombinant AoEH4 by PNGase F Asparagus officinalis

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.80 native enzyme 700fold from roots by anion exchange chromatography, gel filtration step 1, hydrophobic interaction chromatography, and 2fold gel filtration step 2. Recombinant enzyme 99fold from Pichia pastoris by anion exchange chromatography and two different steps of gel filtration Asparagus officinalis
3.2.1.80 native enzyme from roots Asparagus officinalis

Source Tissue

EC Number Source Tissue Comment Organism Textmining
3.2.1.80 root
-
 Asparagus officinalis
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.2.1.80 15
-
 purified recombinant enzyme, pH 5.0, 30°C Asparagus officinalis
3.2.1.80 25.9
-
 purified native enzyme, pH 5.0, 30°C Asparagus officinalis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.80 1-kestose + H2O low activity Asparagus officinalis beta-D-fructose + sucrose
-
 ?
3.2.1.80 1F,6G-di-beta-D-fructofuranosylsucrose + H2O 1 and 6G-kestotetraose, 57-69% activity compared to neokestose Asparagus officinalis ?
-
 ?
3.2.1.80 6-kestose + H2O low activity Asparagus officinalis beta-D-fructose + sucrose
-
 ?
3.2.1.80 6G(1-beta-D-fructofuranosyl)2sucrose + H2O i.e. 1,6G-kestotetraose, 14-15% activity compared to neokestose Asparagus officinalis ?
-
 ?
3.2.1.80 fructosylnystose + H2O 35-38% activity compared to neokestose Asparagus officinalis ?
-
 ?
3.2.1.80 inulin + H2O 24-26.5% activity compared to neokestose Asparagus officinalis ?
-
 ?
3.2.1.80 inulin + H2O and hydrolysis of inulin-type fructans Asparagus officinalis ?
-
 ?
3.2.1.80 additional information fructan 6G&1-exohydrolase (6G&1-FEH) hydrolyzes the terminal beta-2,6 and beta-2,1 fructosyl linkages of fructan, substrate specificity, overview. Levan (from Erwinia herbicola) and sucrose are poor substrates. Hydrolysis of inulin-type fructans Asparagus officinalis ?
-
 ?
3.2.1.80 additional information fructan 6G&1-exohydrolase (6G&1-FEH) hydrolyzes the terminal beta-2,6 and beta-2,1 fructosyl linkages of fructan, substrate specificty, overview Asparagus officinalis ?
-
 ?
3.2.1.80 neokestose + H2O preferred substrate Asparagus officinalis ?
-
 ?
3.2.1.80 nystose + H2O 40-50% activity compared to neokestose Asparagus officinalis ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.80 dimer 2 * 63310, sequence calculation, 2 * 59925, recombinant mature enzyme, sequence calculation, 2 * 76000, glycosylated native enzyme, SDS-PAGE, 2 * 60000, deglycosylated recombinant enzyme, SDS-PAGE Asparagus officinalis

Synonyms

EC Number Synonyms Comment Organism
3.2.1.80 6G&1-FEH
-
 Asparagus officinalis
3.2.1.80 aoeh4a
-
 Asparagus officinalis
3.2.1.80 aoeh4b
-
 Asparagus officinalis
3.2.1.80 fructan 6G&1-exohydrolase
-
 Asparagus officinalis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.80 30
-
 assay at Asparagus officinalis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.80 5
-
 assay at Asparagus officinalis

pI Value

EC Number Organism Comment pI Value Maximum pI Value
3.2.1.80 Asparagus officinalis sequence calculation
-
 5.12

Expression

EC Number Organism Comment Expression
3.2.1.80 Asparagus officinalis enzyme expression increases during low-temperature storage at 2°C from 0 to 9 days, and subsequently decreases. The expression also increases during harvest time from 14 to 70 days up

General Information

EC Number General Information Comment Organism
3.2.1.80 evolution the enzyme belongs to the glycosyl hydrolase family 32, GH32 Asparagus officinalis
3.2.1.80 additional information AoEH4b contains three important amino acid residues, Asp57, Asp182, and Glu236 that are essential for enzymatic reaction catalysis and are conserved in the GH32 family Asparagus officinalis
3.2.1.80 physiological function unlike other 1-FEHs, the enzyme shows minimal hydrolysis of 1-kestose but efficiently hydrolyzes neokestose and inulin-type fructans. The carbohydrates in storage Asparagus officinalis roots are inulin- and inulin neoseries-type fructans Asparagus officinalis
3.2.1.80 physiological function unlike other 1-FEHs, these enzymes shows minimal hydrolysis of 1-kestose but efficiently hydrolyzes neokestose and inulin-type fructans. The carbohydrates in storage Asparagus officinalis roots are inulin- and inulin neoseries-type fructans Asparagus officinalis