5'-Atp → (R)-3-hydroxyoctanoyl-CoA

HRID 490024

반응식

HRID 490024 반응식
P(OC[C@H]1O[C@@H](N2C3N=CN=C(N)C=3N=C2)[C@H](O)[C@@H]1O)(OP(OP(O)(O)=O)(O)=O)(=O)O.[Mg+2].[Cl-].[Cl-].[C@@H:35]1([N:73]2[C:82]3[N:81]=[CH:80][N:79]=[C:77]([NH2:78])[C:76]=3[N:75]=[CH:74]2)[O:72][C@H:44]([CH2:45][O:46][P:47]([O:50][P:51]([O:54][CH2:55][C:56]([C@H:59]([C:61]([NH:63][CH2:64][CH2:65][C:66]([NH:68][CH2:69][CH2:70][SH:71])=[O:67])=[O:62])[OH:60])([CH3:58])[CH3:57])([OH:53])=[O:52])([OH:49])=[O:48])[C@@H:38]([O:39][P:40]([OH:43])([OH:42])=[O:41])[C@H:36]1[OH:37].[OH:83][C@H:84]([CH2:89][CH2:90][CH2:91][CH2:92][CH3:93])[CH2:85][C:86]([O-])=[O:87]>C(O)C(N)(CO)CO.Cl>[OH:83][C@H:84]([CH2:89][CH2:90][CH2:91][CH2:92][CH3:93])[CH2:85][C:86]([S:71][CH2:70][CH2:69][NH:68][C:66](=[O:67])[CH2:65][CH2:64][NH:63][C:61](=[O:62])[C@H:59]([OH:60])[C:56]([CH3:58])([CH3:57])[CH2:55][O:54][P:51]([OH:53])(=[O:52])[O:50][P:47]([OH:49])(=[O:48])[O:46][CH2:45][C@H:44]1[O:72][C@@H:35]([N:73]2[C:82]3[N:81]=[CH:80][N:79]=[C:77]([NH2:78])[C:76]=3[N:75]=[CH:74]2)[C@H:36]([OH:37])[C@@H:38]1[O:39][P:40]([OH:43])([OH:42])=[O:41])=[O:87]

참가 물질

6

반응물 4

HCID 59575'-AtpHCID 87642Coenzyme AHCID 5360315Magnesium ChlorideHCID 86308678(R)-3-hydroxyoctanoate

생성물 1

HCID 11966193(R)-3-hydroxyoctanoyl-CoA

시약·촉매·용매 1

HCID 93573Tromethamine hydrochloride용매

실험 절차

(R)-3-hydroxyoctanoyl-CoA was synthesized according to Rehm B. H. A., Kruger N., Steinbuchel A., Journal of Biological Chemistry, 273, p. 24044-24051, 1998 with slight changes as described below. Acyl-CoA synthetic enzyme (available from Sigma Co.) was dissolved in a Tris-HCl buffer (50 mM, pH 7.5) containing 2 mM ATP, 5 mM MgCl2, 2 mM coenzyme A, and 2 mM (R)-3-hydroxyoctanoate to concentration of 0.1 mU/μl. The mixture was retained in a warm bath at 37° C. and was sampled suitably to analyze progress of a reaction by HPLC. After the enzyme reaction was terminated by adding sulfuric acid to the sampled reaction solution to concentration of 0.02 N, the unreacted substrate (R)-3-hydroxyoctanoate was removed by extraction with n-heptane. The HPLC analysis employed RP18 column (nucleosil C18, 7 μm, Knauser), and elusion was conducted under a linear concentration slope of acetonitrile, using a 25 mM phosphoric acid buffer (pH 5.3) as a moving phase. A thioester compound produced through the enzymatic reaction was detected by monitoring an absorption spectrum of 200 to 500 nm using a diode array detector. (R)-3-hydroxy-5-phenylvaleryl CoA and (R)-3-hydroxy-5-(4-fluorophenyl)valeryl CoA were synthesized in a similar manner.

후처리

  1. customThe mixture was retained in a warm bath at 37° C.
  2. aliquotwas sampled suitably
  3. customprogress of a reaction by HPLC
  4. customAfter the enzyme reaction
  5. customwas terminated
  6. additionby adding sulfuric acid
  7. customto the sampled reaction solution to concentration of 0.02 N
  8. customthe unreacted substrate (R)-3-hydroxyoctanoate was removed by extraction with n-heptane
  9. concentrationwas conducted under a linear concentration slope of acetonitrile

출처 및 근거

ORD 기록
ord-445dc61e1b1b4b809f0bf94b3b6c9479
출처 데이터셋
uspto-grants-2009_05
특허 번호
US07527809B2

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