X 100 → Palmitic Acid

HRID 2259920Notes

Reaction equation

HRID 2259920 reaction equation
C(O)[C:2](N)([CH2:5]O)[CH2:3][OH:4].Cl.SC[C@H:12]([C@@H:14]([CH2:16]S)O)[OH:13].CCC(CO[C:25](C(N(CC[NH+](C)C)C)=O)([C:32]1[CH:37]=[CH:36][CH:35]=[CH:34][CH:33]=1)[C:26]1[CH:31]=[CH:30][CH:29]=[CH:28][CH:27]=1)CC.[Cl-].[Li+].[Cl-].[Mg+2].[Cl-].[Cl-]>>[CH3:33][CH2:32][CH2:25][CH2:26][CH2:27][CH2:28][CH2:29][CH2:5][CH2:2][C:3]([OH:13])=[O:4].[CH3:37][CH2:36][CH2:35][CH2:34][CH2:33][CH2:32][CH2:25][CH2:26][CH2:27][CH2:28][CH2:29][CH2:30][CH2:31][CH2:16][CH2:14][C:12]([OH:4])=[O:13]

Participants

7

Reactant 5

HCID 19153X 100HCID 93573Tromethamine hydrochlorideHCID 433294Lithium ChlorideHCID 446094D-1,4-DithiothreitolHCID 5360315Magnesium Chloride

Product 2

HCID 985Palmitic AcidHCID 2969Decanoic Acid

Procedure

The C10:0-ACP and C16:0-ACP substrates are synthesized enzymatically and are purified by the procedure of Rock et al. (Methods in Enzymol. (1981) 72:397-403). The acyl-ACP substrates for the β-ketoacyl synthase I and II assays require no radiolabel, but sufficient 14C or 3H may be included to monitor the purification after the enzymatic synthesis. The yield of acyl-ACP is also monitored by the filter assay method described by Rock et al. (supra). The synthetic reaction includes, in a 10 ml volume, 4.8 μmol palmitic or decanoic free fatty acid, 800,000 cpm [9,10-3H]palmitic acid (NEN Research Products (Dupont), Boston, Mass., specific activity 28.5 Ci/mmol) or [1-14C]decanoic acid (ICN Biomedicals, Inc, Costa Mesa, Calif., specific activity 3.6 Ci/mol), 100 mM Tris-HCl pH 8.0, 5 mM adenosine 5'-triphosphate, 2 mM dithiothreitol, 2% Triton X-100 (Boehringer Mannheim Biochemicals, Indianopolis, Ind.), 400 mM LiCl, 10 mM MgCl2, 15 mg ACP and 3 units acyl-ACP synthetase. The mix is incubated at 37° C. for three hours and may be stored at room temperature for about 15 hours or at -20° C. until purification.

Workup

  1. customare purified by the procedure of Rock et al. (Methods in Enzymol. (1981) 72:397-403)
  2. customthe purification
  3. customafter the enzymatic synthesis
  4. customThe synthetic reaction
  5. custommay be stored at room temperature for about 15 hours or at -20° C. until purification

Sources & evidence

ORD record
ord-11190c98335d4fa09050bf5fd707b97b
Source dataset
uspto-grants-1995_12
Patent number
US05475099

Reaction notes

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