Information on EC 1.1.1.292 - 1,5-anhydro-D-fructose reductase (1,5-anhydro-D-mannitol-forming)

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1,5-anhydro-D-fructose reductase (ambiguous)

AF reductase

AFR

AKR1E1

NADPH dependent 1,5-anhydro-D-fructose reductase

additional information

1,5-anhydrofructose degradation

1,5-anhydro-D-mannitol + NADP+ = 1,5-anhydro-D-fructose + NADPH + H+

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MetaCyc

1,5-anhydro-D-mannitol:NADP+ oxidoreductase

This enzyme is present in some but not all Rhizobium species and belongs in the GFO/IDH/MocA protein family [2]. This enzyme differs from hepatic 1,5-anhydro-D-fructose reductase, which yields 1,5-anhydro-D-glucitol as the product (see EC 1.1.1.263). In Sinorhizobium morelense, the product of the reaction, 1,5-anhydro-D-mannitol, can be further metabolized to D-mannose [1]. The enzyme also reduces 1,5-anhydro-D-erythro-hexo-2,3-diulose and 2-ketoaldoses (called osones), such as D-glucosone (D-arabino-hexos-2-ulose) and 6-deoxy-D-glucosone. It does not reduce common aldoses and ketoses, or non-sugar aldehydes and ketones [1].

206138-19-4

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1,5-anhydro-D-fructose + NADH + H+

1,5-anhydro-D-mannitol + NAD+

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r

1,5-anhydro-D-fructose + NADPH + H+

1,5-anhydro-D-mannitol + NADP+

5 entries

1,5-anhydro-D-glucitol + NADP+

?

1,5-anhydro-D-mannitol + NADP+

1,5-anhydro-D-fructose + NADPH

4 entries

6-deoxy-D-glucosone + NADPH + H+

6-deoxy-D-mannose + NADP+

9,10-phenanthrenequinone + NADPH

? + NADP+

butane-2,3-dione + NADPH

acetoin + NADP+

D-allosone + NADPH + H+

D-altrose + NADP+

D-glucosone + NADPH + H+

D-mannose + NADP+

D-xylosone + NADPH + H+

D-lyxose + NADP+

17% of the activity with 1,5-anhydro-D-fructose

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?

additional information

?

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1,5-anhydro-D-fructose + NADPH + H+

1,5-anhydro-D-mannitol + NADP+

1,5-anhydro-D-mannitol + NADP+

1,5-anhydro-D-fructose + NADPH

NADH

wild-type enzyme and mutant enzymes S10G, S33D, K94G, D176A, H180A and G206I shows no activity with NADH. Mutant enzymes A13G, S10G/A13G and S33D/A13G are active with NADH

NADP+

NADPH

3 entries

additional information

peptide FPPQSV is absolutely required for induction of the enzyme via AliB-like ORF 2, no activity without the peptide

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1.02 - 49

1,5-Anhydro-D-fructose

15 entries

11

D-glucosone

pH 6.5, 30Â°C

1.1 - 1.2

NADH

5 entries

0.02 - 1

NADPH

9 entries

1.3 - 1300

1,5-Anhydro-D-fructose

16 entries

63.2

D-glucosone

pH 6.5, 30Â°C

288.9

native enzyme

484

recombinant enzyme expressed in Escherichia coli

6.5

-

7

5.2 - 8.8

about 50% of maximal activity at pH 5.5 and at pH 8.8

25

assay at

37

assay at

4.3

isoelectric focusing

i.e. Ensifer adhaerens

667264, 667698

SwissProt

Ensifer adhaerens S-30.7.5

i.e. Ensifer adhaerens

SwissProt

C57BL/6J mice

UniProt

Mus musculus C57/BL6J

C57BL/6J mice

685692

Q9DCT1

UniProt

Sinorhizobium meliloti

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739808

Q92KZ3

UniProt

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UniProt

Streptococcus pneumoniae 110.58

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UniProt

physiological function

Sinorhizobium meliloti 1021

4koa, download, 3D-view

SCOPe (4koa)

CATH (4koa)

Q92KZ3

35100

1 * 35100, MALDI-TOF-MS

38200

gel filtration

40000

1 * 40000, SDS-PAGE

monomer

hanging drop vapor diffusion method, enzyme crystallized in complex with the cofactor NADP(H) and its structure is determined to 2.2 A resolution using selenomethionine single-wavelength anomalous dispersion

in complex with NADP(H), to 1.93 A resolution. The structure displays an empty substrate-binding, showing an open conformation of the enzyme state shortly after the release of product, presumably with bound oxidized cofactor NADP+. Amino-acid residues Lys94, His151, Trp162, Arg163, Asp176 and His180 are involved in substrate binding, catalysis or product release. The side chain of Lys94 may function as a molecular switch

Sinorhizobium meliloti

H180A

kcat/Km for reaction with 1,5-anhydro-D-fructose and NADPH is 61.9fold lower than wild type value

S176A

kcat/Km for reaction with 1,5-anhydro-D-fructose and NADPH is 1001fold lower than wild type value

A13G

kcat/Km for reaction with 1,5-anhydro-D-fructose and NADPH is 1.8fold higher than wild type value. Mutant enzyme shows activity with NADH as cofactor

S10G

kcat/Km for reaction with 1,5-anhydro-D-fructose and NADPH is 1.3fold higher than wild type value

S33D

no activity

K94G

kcat/Km for reaction with 1,5-anhydro-D-fructose and NADPH is 137fold lower than wild type value

G206I

kcat/Km for reaction with 1,5-anhydro-D-fructose and NADPH is 1.4fold lower than wild type value

S10G/A13G

kcat/Km for reaction with 1,5-anhydro-D-fructose and NADPH is 2fold higher than wild type value. Mutant enzyme shows activity with NADH as cofactor

S33D/A13G

kcat/Km for reaction with 1,5-anhydro-D-fructose and NADPH is 84fold lower than wild type value. Mutant enzyme shows activity with NADH as cofactor

-20Â°C or 0Â°C, 50 days, 50% loss of activity

native and recombinant enzyme

recombinant enzyme 17.6fold from Escherichia coli strain BL21 (DE3) by anion exchange and adsorption chromatography

Mus musculus

Q9DCT1

685692

AKR1E1 gene, cloning from liver RNA, DNA and amino acid sequence determination, analysis, and comparison, expression in Escherichia coli strain BL21 (DE3)

Mus musculus

Q9DCT1

685692

expression in Escherichia coli

expresssion in Escherichia coli

Sinorhizobium meliloti

Q92KZ3

739808

gene afr, quantitative real-time RT-PCR enzyme expression analysis

overexpression in Escherichia coli

peptide FPPQSV induces the expression of 1,5-anhydro-D-fructose (AF). The peptide is found in Prevotella species, resulting in enhanced colonization, proteomic analysis is performed by LC-MS/MS on wild-type strain 110.58 and mutant DELTAORF 2 in the presence and absence of AliB-like ORF 2 peptide ligand FPPQSV