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Literature summary for 3.2.1.26 extracted from

  • Linde, D.; Macias, I.; Fernandez-Arrojo, L.; Plou, F.J.; Jimenez, A.; Fernandez-Lobato, M.
    Molecular and biochemical characterization of a beta-fructofuranosidase from Xanthophyllomyces dendrorhous (2009), Appl. Environ. Microbiol., 75, 1065-1073.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.2.1.26

Application

Application Comment Organism
food industry producing short-chain fructooligosaccarides as functional food ingredients Phaffia rhodozyma

Cloned(Commentary)

Cloned (Comment) Organism
S. cervisiae SEY 2101 transformed with pVT103-L plasmid containing invertase fragment Phaffia rhodozyma

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4
-
 sucrose pH 5.5, 42°C, alpha-D-glucopyranosyl-(1,2)-beta-D-fructofuranose Phaffia rhodozyma
4.5
-
 1-kestose pH 5.5, 42°C, alpha-D-glucopyranosyl-(1,2)-beta-D-fructofuranosyl-(1,2)-beta-D-fructofuranose Phaffia rhodozyma

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
66000
-
 after treatment with PNGase F representing the unglydosylated enzyme, thus N-linked oligosaccharides appear to represent about 60% (the other part) of the total protein mass Phaffia rhodozyma
160000
-
 SDS-PAGE and amino acid sequencing Phaffia rhodozyma
200000
-
 smear band in activity-stained gel, probably the about 270 kDa glycosylated functional active homodimer Phaffia rhodozyma

Organism

Organism UniProt Comment Textmining
Phaffia rhodozyma B8YJM2 strain ATCC MYA-131
-

Purification (Commentary)

Purification (Comment) Organism
concentration through a 30000MWCO PES, dialyzation in 20 mM sodium phosphate, pH 7, purification with DEAE-Sephacel chormatography column, elution with 0-0.2 M NaCl gradient, invertase active fractions pooled, dialyzed in sodium acetate buffer, pH 5, applied to a DEAE-Sephacel chromatography volumn and eluted Phaffia rhodozyma

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
 no activity with substrates: maltose (alpha-D-glucopyranosyl-(1,4)-D-glucopyranose), lactose (beta-D-galactopyranosyl-(1,4)-D-glucopyranose), leucrose (alpha-D-glucopyranosyl-(1,5)-D-fructofuranose) Phaffia rhodozyma
90
-
 palatinose as substrate Phaffia rhodozyma
220
-
 nystose as substrate Phaffia rhodozyma
1200
-
 1-kestose as substrate Phaffia rhodozyma
5200
-
 sucrose as substrate Phaffia rhodozyma

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
alpha-D-glucopyranosyl-(1,2)-beta-D-fructofuranose + H2O sucrose, also used to measure transfructosylating activity at high substrate concentration, 0.2 M sodium acetate buffer, pH 5.6, 50°C Phaffia rhodozyma D-glucose + D-fructose + ? products are mainly glucose and fructose, but also neokestose, 1-kestose, and nystose are built ?
alpha-D-glucopyranosyl-(1,2)-beta-D-fructofuranosyl-(1,2)-beta-D-fructofuranose + H2O 1-kestose, about 25% activity compared to sucrose, 0.2 M sodium acetate buffer, pH 5.6, 50°C Phaffia rhodozyma D-glucose + ?
-
 ?
alpha-D-glucopyranosyl-(1,2)-beta-D-fructofuranosyl-(1,2)-beta-D-fructofuranosyl-(1-2)-beta-D-fructofuranose + H2O nystose, about 4% activity compared to sucrose, 0.2 M sodium acetate buffer, pH 5.6, 50°C Phaffia rhodozyma D-glucose + ?
-
 ?
alpha-D-glucopyranosyl-(1,6)-D-fructofuranose + H2O palatinose, about 2% activity compared to sucrose, 0.2 M sodium acetate buffer, pH 5.6, 50°C Phaffia rhodozyma alpha-D-glucopyranose + D-fructofuranose
-
 ?

Synonyms

Synonyms Comment Organism
beta-fructofuranosidase
-
 Phaffia rhodozyma
yeast invertase
-
 Phaffia rhodozyma

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
65 70
-
 Phaffia rhodozyma

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
25 90 less than 10% activity at 25 and above 80°C, 100% activity at 65-70°C, about 90% activity at 60 and 75°C, about 50% activity at 50-55°C, 10-40% activity at 35-45°C Phaffia rhodozyma

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
40 85 minor activity loss after 40 days at 40-50°C, 50% retained after 1 d at 60°C, inactivation after 10 min at 85°C, pre-incubation for 10-120 min at 60-85°C: 50% activity loss from 66-71°C, 50 mM sodium phosphate buffer, pH 5.5, remaining activity measured at 42°C Phaffia rhodozyma

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.5
-
 1-kestose alpha-D-glucopyranosyl-(1,2)-beta-D-fructofuranosyl-(1,2)-beta-D-fructofuranose Phaffia rhodozyma
5.7
-
 sucrose alpha-D-glucopyranosyl-(1,2)-beta-D-fructofuranose Phaffia rhodozyma

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5.5
-
 highest activity at pH 5.5-6.0 Phaffia rhodozyma

pH Range

pH Minimum pH Maximum Comment Organism
3 8 highest activity at pH 5.5-6.0, more than 75% at pH 5, and 6.5-7, more than 50% at pH 4, about 25% at pH 8, about 10% at pH 3, substrate sucrose, buffers at 100 mM, pH 3-4: citric acid-sodium citrate, pH 4-7: Na2HPO4-NaH2PO4, pH 7-8: Tris-HCl Phaffia rhodozyma