4-(2-Hydroxyethyl)-1-piperazineethanesulfonic acid → N-Acetyl-D-Glucosamine

HRID 2252515

반응식

HRID 2252515 반응식
C1N(CCO)CCN(CCS(O)(=O)=O)C1.[OH:16][C@H:17]1[O:24][C@H:23]([CH2:25][OH:26])[C@H:21]([OH:22])[C@H:19]([OH:20])[C@H:18]1[NH:27][C:28]([CH3:30])=[O:29].CC(N[C@H]1[C@@H](OP(OP(OC[C@H]2O[C@@H](N3C(=O)NC(=O)C=C3)[C@H](O)[C@@H]2O)(O)=O)(O)=O)O[C@H](CO)[C@@H](O)[C@@H]1O)=O>C(#N)C>[C:28]([NH:27][C@@H:18]1[C@@H:19]([OH:20])[C@H:21]([OH:22])[C@@H:23]([CH2:25][OH:26])[O:24][CH:17]1[OH:16])(=[O:29])[CH3:30]

참가 물질

5

반응물 3

HCID 238314-(2-Hydroxyethyl)-1-piperazineethanesulfonic acidHCID 84265N-acetyl-alpha-D-galactosamineHCID 445675Uridine-Diphosphate-N-Acetylglucosamine

생성물 1

HCID 439174N-Acetyl-D-Glucosamine

시약·촉매·용매 1

HCID 6342Acetonitrile용매

실험 절차

The reaction solution contained 50 mM HEPES buffer, 10 mM MnCl2, 0.1% Triton-CF54, 1 mM pNp-α-GalNAc and 1.5 mM UDP-GlcNAc, to which 5 μl of β3Gn-T6 was added, so as to have a total volume of 20 μl. The reaction was carried out at 37° C. for 16 hours. After completion of the reaction, analysis was carried out using HPLC. The column used was Mightysil RP-18, 250×4 mm, the solvent was acetonitrile: H2O=10:90, and the detection was carried out in terms of the absorption at 210 nm. As standard samples, GlcNAcβ1-3GalNAcα-pNp (core 3 structure) and GlcNAcβ1-6GalNAcα-pNp (core 6 structure) were used (SIGMA). As a result, as for the standard samples, GalNAcα-pNp was eluted at 23.5 minutes, GlcNAcβ1-3GalNAcα-pNp was eluted at 18.6 minutes and GlcNAcβ1-6GalNAcα-pNp was eluted at 20 minutes. As for the β3Gn-T6 solution, the peak at 23.5 minutes (GalNAcα-pNp) and the peak at 18.6 minutes (GlcNAcβ1-3GalNAcα-pNp) were observed. Thus, β3Gn-T6 is a novel enzyme having an activity to transfer N-acetylglucosamine to a non-reducing terminal of N-acetylgalactosaminyl group through the β1,3-linkage.

후처리

  1. additionto which 5 μl of β3Gn-T6 was added
  2. customAfter completion of the reaction, analysis
  3. customH2O=10:90, and the detection was carried out in terms of the absorption at 210 nm
  4. washAs a result, as for the standard samples, GalNAcα-pNp was eluted at 23.5 minutes
  5. washGlcNAcβ1-3GalNAcα-pNp was eluted at 18.6 minutes
  6. washGlcNAcβ1-6GalNAcα-pNp was eluted at 20 minutes
  7. waitAs for the β3Gn-T6 solution, the peak at 23.5 minutes (GalNAcα-pNp)
  8. waitthe peak at 18.6 minutes (GlcNAcβ1-3GalNAcα-pNp) were observed

출처 및 근거

ORD 기록
ord-f05fc662d1bb4c4a8e415328838b80e4
출처 데이터셋
uspto-grants-2008_01
특허 번호
US07323324B2

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