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3.2.1.26: beta-fructofuranosidase

This is an abbreviated version!
For detailed information about beta-fructofuranosidase, go to the full flat file.

Word Map on EC 3.2.1.26

Reaction

raffinose
+
H2O
=
beta-D-fructofuranose
+
6-O alpha-D-galactopyranosyl-alpha-D-glucopyranose

Synonyms

A/N-InvG, acid invertase, Acid sucrose-6-phosphate hydrolase, acINV, AFR529Wp, AI, AIV, alkaline invertase, alkaline invertase InvA, alkaline invertase InvB, alkaline/neutral invertase, Atbetafruct4, b-fructofuranosidase, beta-(1-2)-fructofuranosidase, beta-D-fructofuranosidase, beta-D-fructofuranosidase fructohydrolase, beta-D-fructofuranoside fructohydrolase, beta-FFase, beta-fructofuranosidase, beta-fructofuranosidase I, beta-fructofuranoside fructohydrolyase, beta-fructosidase, beta-h-fructosidase, BfrA, BlsacA, Bmsuc1, cell wall invertase, cell wall invertase 4, cell wall-bound invertase, cell wall-bound ivertase, cell-wall invertase, cell-wall invertase 1, CIN, Cin1, CIN12, Cin5, CINV1, CscA, CWI, CWIN, CWIN1, Cwinv-1, cwINV1, cytosolic invertase, EINV1, EINV2, EINV3, EINV4, FCWI, Ffase, FFase I, Ffh, fructofuranosidase, beta-, fructosylinvertase, glucosucrase, INAC-INV, INCW2, INV, Inv-CW, INV-E, Inv-V, INV1p, INV2, INVA, INVB, invertase, invertase 1, invertase 2, Invertase E1, invertase SUC2, invertin, lbbetafruct2, lbbetafruct3, Lin5, Lin6, Lin7, Lyc e 2.01, Lyc e 2.02, maxinvert L 1000, More, neutral invertase, NI, Nin88, OsCIN1, OsCIN2, OsCIN3, Protein B46, re-INVB, SacA, saccharase, SAI, soluble acid invertase, Suc2, SucB, sucrase, Sucrase E1, Sucrose-6-phosphate hydrolase, TIV-1, Uninv, uninv2, Vacuolar invertase, vacuolar invertase 1, vacuolar invertase CvINV, vacuolar invertase NvINV, VIN, VIN1, VIN2, Vinv-1, VINV1, VINV2, VINV3, yeast invertase

ECTree

     3 Hydrolases
         3.2 Glycosylases
             3.2.1 Glycosidases, i.e. enzymes that hydrolyse O- and S-glycosyl compounds
                3.2.1.26 beta-fructofuranosidase

Engineering

Engineering on EC 3.2.1.26 - beta-fructofuranosidase

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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
F348L
wild-type activity
L201M
wild-type activity
N251C
wild-type activity
S457N
wild-type activity
V276Y
no activity
W343Y
wild-type activity
W440Y
almost doubled transglycosylation activity
Y394F
no activity
C294Y
-
i.e. mutant sicy-192. Mutation leads to inhibition of cotyledon greening by treatments with sugars such as sucrose, glucose, and fructose. The greening of cotyledons in the knock-out INV-E lines is not inhibited by treatment with the sugars. A recombinant INV-E:C294Y protein has the same enzymatic activity as a recombinant INV-E protein, suggesting that the Cys-294 residue of INV-E is important for its functions in the chloroplasts. On treatment with sucrose, the expression of photosynthesis-related genes is weaker in seedlings of mutant plants than wild-type seedlings, whereas the activity of nitrate reductase is stronger in the mutant plants than wild-type plants
D239A
-
shows a 6fold increase in KM and 1-kestose exohydrolase activity
D23A
-
inactive
E203A
-
inactive
E203Q
-
inactive
S547A
the mutant is unable to interact with 14-3-3 protein
C317A
-
2.5fold invertase activity increase, 20.7% activity with substrate raffinose compared to wild-type
D135E
-
no invertase activity
D135N
-
no invertase activity
D260E
-
no invertase activity
D260N
-
no invertase activity
E316A
-
no invertase activity
E316Q
-
67.3% invertase activity retained, 15.5% activity with substrate raffinose compared to wild-type
R259K
-
no invertase activity
Rs59L
-
no invertase activity
W159F
-
4.7fold invertase activity increase, 41% activity with substrate raffinose compared to wild-type
W159L
-
11.7% invertase activity retained, 33.2% activity with substrate raffinose compared to wild-type
D181A
mutant enzyme shows no detectable activity
D63A
catalytic efficiency (kcat/Km) of the mutant enzyme is 1.6% compared to the catalytic efficiency of the wild-type enzyme
E234A
catalytic efficiency (kcat/Km) of the mutant enzyme is 1.8% compared to the catalytic efficiency of the wild-type enzyme
R429A
catalytic efficiency (kcat/Km) of the mutant enzyme is 9% compared to the catalytic efficiency of the wild-type enzyme
R429N
catalytic efficiency (kcat/Km) of the mutant enzyme is 20% compared to the catalytic efficiency of the wild-type enzyme
R430A
mutant enzyme shows no detectable activity
R430N
catalytic efficiency (kcat/Km) of the mutant enzyme is 16% compared to the catalytic efficiency of the wild-type enzyme
R205G
-
mutant Oscyt-inv1 shows abnormal phenotypes with short primary, adventitious, and lateral roots (reduced cell lengths, disarray of cells but radial pattern conserved), floral transition delay, low seed setting rate, low pollen fertility
D22N
-
activity bleow detection limit with sucrose. Increase in transfructosylating activity
F82W
-
71% of wild-type activity
K185R
mutant enzyme K185R shows higher stability than wild-type enzyme at 40°C-50°C. These results suggest that the deubiquitination of K185 slightly affects the thermal stability of SUC2
N21S
-
104% of wild-type activity with sucrose. Large increase in transfructosylating activity
N228A
-
138% of wild-type activity
N228A/N21S
-
20% of wild-type activity
N228R
-
23% of wild-type activity, no transfructosylating activity
N228R/N21S
-
4% of wild-type activity
N24S
-
17% of wild-type activity with sucrose. Large increase in transfructosylating activity
P205V
-
112% of wild-type activity with sucrose. Large increase in transfructosylating activity
P205V/W19Y/N21S
-
3% of wild-type activity with sucrose. Large increase in transfructosylating activity
W19Y
-
175% of wild-type activity with sucrose. Increase in transfructosylating activity
W19Y/N21S
-
41% of wild-type activity with sucrose. Large increase in transfructosylating activity
W19Y/N21S/N24S
-
2% of wild-type activity with sucrose. Large increase in transfructosylating activity
W19Y/N24S
-
8% of wild-type activity with sucrose. Large increase in transfructosylating activity
W287Y
-
4% of wild-type activity with sucrose. Increase in transfructosylating activity
W291Y
-
32% of wild-type activity with sucrose. Increase in transfructosylating activity
K185R
-
mutant enzyme K185R shows higher stability than wild-type enzyme at 40°C-50°C. These results suggest that the deubiquitination of K185 slightly affects the thermal stability of SUC2
-
D50A
-
no catalytic activity
E230A
-
no catalytic activity
N142Y
-
large decrease in activity against nystose
N254A
-
large decrease in activity against nystose
Q176E
-
moderate change in catalytic activity
Q176S
-
moderate change in catalytic activity
Q228V
-
strikingly large reduction in the catalytic efficiency of sucrose , inulin and especially nystose
Q435A
-
significant decrease in catalytic efficiency against inulin
S281I
-
strikingly large reduction in the catalytic efficiency of sucrose , inulin and especially nystose
Y462A
-
significant decrease in catalytic efficiency against inulin
N119D
the mutant shows reduced activity compared to the wild type enzyme
N184D
the mutant shows reduced activity compared to the wild type enzyme
N516D
the mutation dramatically affects the folding of the protein. The mutant is inactive
N52D
the mutant shows reduced activity compared to the wild type enzyme
N52D/N119D/N184D
the mutant shows reduced activity compared to the wild type enzyme
N52D/N119D/N184D/N516D
the mutations dramatically affect the folding of the protein. The mutant is inactive
E190A
site-directed mutagenesis, inactivated mutant
E190D
site-directed mutagenesis, inactivated mutant
additional information