Beteiligte Stoffe
| Rolle | Name | HCID | Einwaage |
|---|
| Edukt | NADPH | HCID 5884 | — |
| Edukt | Acetonitrile | HCID 6342 | — |
| Edukt | 2,4,6/3,5-Pentahydroxycyclohexanone | HCID 439294 | — |
| Edukt | HCID 178521643 | HCID 178521643 | — |
| Lösungsmittel | Tris(hydroxymethyl)aminomethane | HCID 6503 | — |
| Produkt | An inositol | HCID 892 | — |
Reaktionsschritte
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 Φ4.6×250 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.
Herkunft & Nachweise
| DOI | 10.6084/m9.figshare.5104873.v1 |
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| Patentnummer | US07745671B2 |
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| Quelldatensatz | uspto-grants-2010_06 |
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| ORD ID | ord-fd7f3b8f3c8d4800814d73eb526368fb |
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| Quellennotiz | uspto-grants-2010_06 |
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