Information on EC 4.2.2.17 - inulin fructotransferase (DFA-I-forming)

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inulin D-fructosyl-D-fructosyltransferase (1,2':2',1-dianhydride-forming)

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inulin fructotransferase (depolymerizing, difructofuranose-1,2':2',1-dianhydride-forming)

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inulin fructotransferase (DFA I-producing)

inulin fructotransferase (DFA-I-producing)

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NBCG_01407

locus name

Noca_0068

UG56_016290

locus name

Produces alpha-D-fructofuranose beta-D-fructofuranose 1,2':2,1'-dianhydride (DFA I) by successively eliminating the diminishing (2->1)-beta-D-fructan (inulin) chain from the terminal D-fructosyl-D-fructosyl disaccharide.

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D-fructosyl-D-fructofuranosyl group transfer

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(2->1)-beta-D-fructan lyase (alpha-D-fructofuranose-beta-D-fructofuranose-1,2':2,1'-dianhydride-forming)

This enzyme, like EC 4.2.2.16 [levan fructotransferase (DFA-IV-forming)] and EC 4.2.2.18 [inulin fructotransferase (DFA-III-forming)] eliminates the fructan chain from the terminal disaccharide leaving a difructose dianhydride. These enzymes have long been known as fructotransferases, so this is retained in the accepted name. Since the transfer is intramolecular, the reaction is an elimination and, hence, the enzyme is a lyase, belonging in EC 4.

125008-19-7

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beta-D-fructofuranosyl-(2->1)-beta-D-fructofuranosyl-(2->1)-beta-D-fructofuranosyl-(2->1)-beta-D-fructofuranosyl-(2->1)-alpha-D-glucopyranoside

di-D-fructofuranose-1,2':2,1'-dianhydride + 1-kestose

4 entries

fructofuranosyl nystose

di-D-fructofuranose-1,2':2,1'-dianhydride + 1-kestose

inulin

3 di-D-fructofuranose-1,2':2,1'-dianhydride + H2O

11 entries

inulin

di-D-fructofuranose-1,2':2,1'-dianhydride + 1-kestose + nystose + fructofuranosyl nystose

inulin

di-D-fructofuranose-1,2':2,1'-dianhydride + sucrose + 1-kestose

4 entries

nystose

di-D-fructofuranose-1,2':2,1'-dianhydride + sucrose

6 entries

additional information

?

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inulin

3 di-D-fructofuranose-1,2':2,1'-dianhydride + H2O

8 entries

Co2+

-

weak

Cu2+

-

weak

Fe3+

-

1 mM, strong

Hg2+

-

1 mM, strong

Ni2+

-

weak

EDTA

-

slight stimulation

2.89 - 4

inulin

114

Nocardioides luteus

A0A1J4N453

substrae inulin, pH 5.5, 55°C

127

-

131

Nocardioides sp.

A1SCS9

substrate inulin, pH 5.5, 50°C

147

E9URP8

substrate inulin, pH 6.0, 45°C

172

-

5.5

4 entries

6

3 entries

3.5 - 8

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10% of maximal activity at pH 3.5, 25% of maximal activity at pH 8.0

4.5 - 7

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pH 4.5: about 50% of maximal activity, pH 7.0: about 50% of maximal activity

6 - 6.5

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more than 69% of maximum activity

40

45

50

55

-

30

-

80% of maximal activity at 30°C, 20% of maximal activity at 65°C

30 - 60

-

30°C: about 70% of maximal activity, 60°C: about 45% of maximal activity

30 - 65

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30°C: about 80% of maximal activity

Arthrobacter

MC12493

644466

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Arthrobacter globiformis S14-3

S14-3

644464, 644465, 644467

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S14-3

644464, 644465, 644467

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Arthrobacter MC12493

MC12493

644466

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Nocardioides sp. ATCC BAA-499

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UniProt

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UniProt

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UniProt

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A1SCS9

UniProt

Paenarthrobacter ureafaciens A51-1

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664404, 664513

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Paenarthrobacter ureafaciens a62-1

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-

-

-

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12500

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gel fitration

37000

-

2 * 37000, SDS-PAGE

38000

-

2 * 38000, SDS-PAGE

39000

-

SDS-PAGE

644465

40000

Arthrobacter

-

SDS-PAGE and gel filtration

644466

41600

-

amino acid analysis

644465

46000

-

gel filtration

50000

gel filtration

58000

E9URP8

gel filtration

60000

-

gel filtration

61000

-

gel filtration

?

Nocardioides luteus

A0A1J4N453

x * 41422, calculated from sequence, x * 41000, SDS-PAGE

dimer

monomer

trimer

homology modelling and molecular docking. Residues D183 and E194 are potential catalytic residues

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molecular modeling of strucuture

A257S

mutation of residue in central tunnel, no significant change of temperature stability

A257S/G281S G281S

mutation of residues in central tunnel, increase in temperature stability by 3 degrees

A257T

mutation of residue in central tunnel, 2.8 degrees decrease of temperature stability

D183A

-

complete loss of activity

D183N

-

complete loss of activity

E194A

-

complete loss of activity

E194N

-

complete loss of activity

G121A/T122L

148.7% of wild-type activity, about 4.5 degrees increase in temperature stability

G236S/G281S/A257S/T313S/A314S

140.5% of wild-type activity, increase in temperature stability by 3.5 degrees

G281S

mutation of residue in central tunnel, 76% of wild-type activity, increase in temperature stability by 3 degrees

G281T

mutation of residue in central tunnel, no significant change of temperature stability

I80A

11.6% of wild-type activity, about 2 degrees increase in temperature stability

I80E

15.7% of wild-type activity, about 1.5 degrees increase in temperature stability

I80S

41.4% of wild-type activity, about 1 degrees increase in temperature stability

D183A

-

complete loss of activity

-

D183N

-

complete loss of activity

-

E194A

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complete loss of activity

-

E194N

-

complete loss of activity

-

3

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stable at 25°C

644467

4

Arthrobacter

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30°C, 30 min., stable

644466

30

-

stable, rapid inactivation above 80°C

70

72.8

melting temperature of mutant A257T

75

75.6

melting temperature of wild-type and mutant A257S

78.8

melting temperature of mutant G236S/G281S/A257S/T313S/A314S

80

3 entries

80.1

melting temperature of mutant G121A/T122L

85

-

pH 5.5, 1 h, complete inactivation

-

-

664404, 664513

partial

-

-

-

644464

expression in Escherichia coli

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747956, 748014

expression in Escherichia coli MV1184