Ampicillin → 2-Benzyl-3-hydroxyglutarate diethyl ester

HRID 2308037Notes

Reaction equation

HRID 2308037 reaction equation
C(C(C(=O)CC([O-])=O)C([O-])=O)C1C=CC=CC=1.O=C[C@@H]([C@H]([C@@H]([C@@H](CO)O)O)O)O.CC1(C)S[C@@H]2[C@H](NC([C@H](N)C3C=CC=CC=3)=O)C(=O)N2[C@H]1C(O)=O.CC(S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O)C.[CH2:69]([O:71][C:72](=[O:89])[CH:73]([CH2:82][C:83]1[CH:88]=[CH:87][CH:86]=[CH:85][CH:84]=1)[C:74](=[O:81])[CH2:75][C:76]([O:78][CH2:79][CH3:80])=[O:77])[CH3:70]>C(OCC)(=O)C.CS(C)=O>[CH2:69]([O:71][C:72](=[O:89])[CH:73]([CH2:82][C:83]1[CH:84]=[CH:85][CH:86]=[CH:87][CH:88]=1)[CH:74]([OH:81])[CH2:75][C:76]([O:78][CH2:79][CH3:80])=[O:77])[CH3:70]

Participants

8

Reactant 5

HCID 6249Ampicillin2×HCID 107526(2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanal3×HCID 656894isopropyl beta-D-thiogalactopyranoside2×HCID 12200242Diethyl 2-benzyl-3-oxopentanedioate3×HCID 220229172-Benzyl-3-ketoglutarate

Product 1

HCID 220229062-Benzyl-3-hydroxyglutarate diethyl ester

Reagents, catalysts & solvents 2

HCID 679Dimethyl SulfoxideSolventHCID 8857Ethyl AcetateSolvent

Procedure

Whole cell reductions of 2-benzyl-3-ketoglutarate were carried out using E. coli cells expressing the gene encoding KRED 1008. Cells were grown overnight at 30 C in 400 mL of a Luria Broth/glucose (2 g/L)/ampicillin (100 mg/L) media. The cells were isolated by centrifugation and re-suspended in 400 mL of a minimal salt (M9) solution containing isopropylthiogalactoside (IPTG, 0.5 mM), glucose (5 g/L), ampicillin (100 mg/L), and 2-benzyl-3-ketoglutarate diethyl ester. In independent experiments, the concentration of 2-benzyl-3-ketoglutarate diethyl ester was varied over the range of 1 gram/liter to 7.5 grams/liter. Cells continued to grow at 30 C after addition of the ketodiester substrate. Samples were removed at regular time intervals and analyzed for product formation using gas chromatography until complete reaction was observed. Although the 2-benzyl-3-ketoglutarate diethyl ester substrate did not completely dissolve in the aqueous buffer, and although no DMSO co-solvent was used, the reduction reactions were complete after 20-40 hours. In reactions that were allowed to proceed for more that 24 hours, another portion of glucose (1-3 g) and IPTG (0.4 mL of a 0.5 M solution) were added. The product 2-benzyl-3-hydroxyglutarate diethyl ester was isolated by removal of the cells by centrifugation and extraction of the reaction mixture with ethyl acetate. The yield of 2-benzyl-3-hydroxyglutarate diethyl ester in all cases was greater than 80%.

Workup

  1. customThe cells were isolated by centrifugation
  2. customSamples were removed at regular time intervals
  3. customuntil complete reaction
  4. custommore that 24 hours

Sources & evidence

ORD record
ord-ce0b3bae8be9427094fe5e7b9b1bb114
Source dataset
uspto-grants-2004_12
Patent number
US06833471B2

Reaction notes

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