NADPH → An inositol

HRID 1019480

Reaktionsgleichung

Reaktionsgleichung von HRID 1019480
[C@H:1]1([OH:12])[CH:7]([OH:8])[C@@H:6]([OH:9])[C@H:5]([OH:10])[C:3](=[O:4])[C@@H:2]1[OH:11].C1N=C(N)C2N=CN([C@@H]3O[C@H](COP(OP(OC[C@H]4O[C@@H](N5C=C(C(N)=O)CC=C5)[C@H](O)[C@@H]4O)(O)=O)(O)=O)[C@@H](O)[C@H]3OP(O)(O)=O)C=2N=1.C(#N)C.[C@@H]1(O)[C@@H](O)[C@H](O)[C@@H](O)[C@H](O)[C@H]1O>C(O)C(N)(CO)CO>[CH:3]1([OH:4])[CH:2]([OH:11])[CH:1]([OH:12])[CH:7]([OH:8])[CH:6]([OH:9])[CH:5]1[OH:10]

Komponenten

6

Edukt 4

HCID 5884NADPHHCID 6342AcetonitrileHCID 4392942,4,6/3,5-PentahydroxycyclohexanoneHCID 178521643HCID 178521643, Verbindung2×

Produkt 1

HCID 892An inositol

Reagenzien, Katalysatoren, Lösungsmittel 1

HCID 6503Tris(hydroxymethyl)aminomethaneLösungsmittel

Versuchsvorschrift

Meanwhile, the enzyme reaction product was measured as follows: 10 mg of scyllo-inosose, 40 mg of NADPH, and 10 U of the enzyme were allowed to react in 1.0 ml of 100 mM Tris buffer (pH 8.0) at 36° C. for 4 hours, and a heat treatment was performed at 80° C. for 10 min, followed by cooling. Then, 100 μl of a strong base cation exchange resin, 100 μl of a strong acid anion exchange resin, and 10 mg of activated carbon were added, and the mixture was stirred and centrifuged. Then, the supernatant was diluted 2-fold, and measurement was performed by HPLC (Shodex Asahipak NH2P-50 4E Φ 4.6×250 mm: Shodex) using an RI detector under the condition of a column temperature of 40° C. and a mobile phase flow rate of 1.5 ml (80% acetonitrile). As a result, the ydgJ gene product was found to have a high scyllo-inositol dehydrogenase activity, and 100% of the product was scyllo-inositol obtained by reduction, while myo-inositol that is an isomer thereof was not detected. As a result, the solution of a recombinant enzyme derived from the ydgJ gene was found to have a high scyllo-inositol dehydrogenase activity, and the gene product was scyllo-inositol dehydrogenase. Meanwhile, in the product obtained by the reduction reaction of scyllo-inosose, only scyllo-inositol was detected, and the enzyme was found to stereospecifically reduce scyllo-inosose into scyllo-inositol. Meanwhile, the sequence of the ydgJ gene is shown in SEQ ID NO: 1, and the amino acid sequence corresponding thereto is shown in SEQ ID NO: 2.

Aufarbeitung

  1. temperatureby cooling
  2. additionThen, the supernatant was diluted 2-fold
  3. customunder the condition of a column temperature of 40° C.

Quellen und Nachweise

ORD-Eintrag
ord-9e498f488fb2463a9a10c11eddcd2f9e
Quelldatensatz
uspto-grants-2014_05
Patentnummer
US08715974B2

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