참여 물질
| 역할 | 이름 | HCID | 투입량 |
|---|
| 반응물 | (3S,4S,5R)-1,3,4,5,6-pentahydroxyhexan-2-one | HCID 92092 | — |
| 반응물 | Tromethamine hydrochloride | HCID 93573 | — |
| 반응물 | (2R,3S,4S,5R)-2,3,4,5,6-pentahydroxyhexanal | HCID 3037556 | — |
| 반응물 | Magnesium Chloride | HCID 5360315 | — |
| 반응물 | Cystein carbazol-sulfuric acid | HCID 22799518 | — |
| 생성물 | D-Arabinose | HCID 66308 | — |
반응 단계
The isomerization of galactose was performed with a mixture of 25 μl of 100 mM galactose and 100 μl of the crude enzyme solution as a substrate at 60° C. for 1 hour. To measure the activity of galactose isomerization, 100 μl of the crude enzyme solution containing 40 mM of galactose as a substrate was mixed with 1 Ml of reaction buffer (50 mM Tris-HCl, pH 7.0), followed by reaction at 65° C. for 20 minutes. At that time, 5 mM of MgCl2 and 1 mM of MnCl2 were added to the reaction mixture. The activity of the isomerase was measured by the cystein-carbazol-sulfuric acid method (Dische, Z., and E. Borenfreund., A New Spectrophotometric Method for the Detection and Determination of Keto Sugars and Trioses, J. Biol. Chem., 192:583-587, 1951). The protein contained in the crude enzyme solution was quantified with a Bradford assay kit (Biorad, U.S.A.). As a result, the isomerase activity was 0.2045±0.0078 (mg-tagatose/mg-protein·h), indicating that the product of galactose isomerization, tagatose, was successfully generated (FIG. 3).
출처 및 근거
| DOI | 10.6084/m9.figshare.5104873.v1 |
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| 특허 번호 | US08137946B2 |
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| 출처 데이터셋 | uspto-grants-2012_03 |
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| ORD ID | ord-b58030edd3e8478883cdca7def9287ef |
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| 출처 메모 | uspto-grants-2012_03 |
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