Application | Comment | Organism |
---|---|---|
food industry | producing short-chain fructooligosaccarides as functional food ingredients | Phaffia rhodozyma |
Cloned (Comment) | Organism |
---|---|
S. cervisiae SEY 2101 transformed with pVT103-L plasmid containing invertase fragment | Phaffia rhodozyma |
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 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 | UniProt | Comment | Textmining |
---|---|---|---|
Phaffia rhodozyma | B8YJM2 | strain ATCC MYA-131 | - |
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 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 | 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 | Comment | Organism |
---|---|---|
beta-fructofuranosidase | - |
Phaffia rhodozyma |
yeast invertase | - |
Phaffia rhodozyma |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
65 | 70 | - |
Phaffia rhodozyma |
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 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 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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5.5 | - |
highest activity at pH 5.5-6.0 | Phaffia rhodozyma |
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 |