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NADPH → HCID 67258465、化合物

HRID 1486229
ノート
印刷プレビュー

反応式

HRID 1486229 の反応式
C1N=C(N)C2N=CN([C@@H]3[O:14][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]3O)C=2N=1.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.O=C[C@@H]([C@H]([C@@H]([C@@H](CO)O)O)O)O.[O-]S([O-])(=O)=O.[Mg+2].[F:111][C:112]1[CH:117]=[CH:116][C:115]([C:118](=[O:136])[CH2:119][CH2:120][CH2:121][C:122]([N:124]2[C@@H:128]([C:129]3[CH:134]=[CH:133][CH:132]=[CH:131][CH:130]=3)[CH2:127][O:126][C:125]2=O)=[O:123])=[CH:114][CH:113]=1>CC(OC)(C)C.C(#N)C.C(O)(C)C>[O:14]=[C:130]1[CH:131]=[CH:132][CH:133]=[CH:134][CH:129]1[C@H:128]1[CH2:127][O:126][CH2:125][N:124]1[C:122](=[O:123])[CH2:121][CH2:120][CH2:119][C:118]([C:115]1[CH:116]=[CH:117][C:112]([F:111])=[CH:113][CH:114]=1)=[O:136]
参加物実験手順後処理出典と根拠反応ノート

参加物

10

反応物 6

HCID 5884NADPHHCID 24083Sulfuric acid magnesium salt (1:1)HCID 107526(2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanal2 回HCID 439153NadhHCID 11187377(S)-1-(4-Fluorophenyl)-5-(2-oxo-4-phenyloxazolidin-3-yl)pentane-1,5-dioneHCID 178435179HCID 178435179、化合物

生成物 1

HCID 67258465HCID 67258465、化合物

試薬・触媒・溶媒 3

HCID 3776Isopropanol溶媒HCID 6342Acetonitrile溶媒HCID 15413tert-Butyl methyl ether溶媒

実験手順

KREDs described in Table 5 of Example 1 are screened using NADH and NADPH as cofactors and glucose dehydrogenase/glucose or isopropylalcohol (“IPA”) as co-factor regeneration system. 100 μl of cell lysate was added to a deep well plate (Costar #3960) containing 25 μl 5 mg/ml Na-NADP (Oriental Yeast) and 2 mM MgSO4 in 100 mM triethanolamine(chloride) (pH7.0), and 125 IA isopropyl alcohol containing 2 g/L (S)-1-(4-Fluoro-phenyl)-5-(2-oxo-4-phenyl-oxazolidin-3-yl)-pentane-1,5-dione. After sealing the plates with aluminum/polypropylene laminate heat seal tape (Velocity 11 (Menlo Park, Calif.), Cat#06643-001), reactions were run for at least 16 hrs at ambient temperature. At the end of the reaction 1 ml acetonitrile (for reversed phase HPLC) or MTBE (for normal phase HPLC) was added per well. Plates were resealed, shaken for 20 minutes, and centrifuged (4000 rpm, 10 min, 4° C.). 200 μl of the organic layer was transferred into a new shallow-well microtiter plate for analysis.

後処理

  1. addition100 μl of cell lysate was added to a deep well plate (Costar #3960)
  2. temperatureheat
  3. customseal tape (Velocity 11 (Menlo Park, Calif.), Cat#06643-001), reactions
  4. waitwere run for at least 16 hrs at ambient temperature
  5. additionwas added per well
  6. customcentrifuged (4000 rpm, 10 min, 4° C.)

出典と根拠

ORDレコード
ord-44857b5b9e014069a38d644e4a6daf46
出典データセット
uspto-grants-2016_08
DOI
10.6084/m9.figshare.5104873.v1
特許番号
US09422530B2

反応ノート

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