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Volumn 21, Issue 2, 2010, Pages 159-162

Optical 2-benzyl-5-hydroxy-4-oxopentanoic acids against carboxypeptidase A: Synthesis, kinetic evaluation and X-ray crystallographic study

Author keywords

2 Benzyl 5 hydroxy 4 oxopentanoic acid; Carboxypeptidase A; X ray crystallographic study

Indexed keywords

EID: 72549095860 PISSN: 10018417 EISSN: None Source Type: Journal
DOI: 10.1016/j.cclet.2009.09.005 Document Type: Article

Times cited : (3)

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- Determination of Ki value. CPA was purchased from Sigma Chemical Co, Allan form, twice crystallized from bovine pancrease, aqueous suspension in toluene) and used without further purification. CPA stock solutions were prepared by dissolving the enzyme in 0.05 mol L-1 Tris/0.5 mol L-1 NaCl, pH 7.5 buffer solution. O-(trans-p-chlorocinnamoyl)-l-β-phenyllactate (Cl-CPL, which was synthesized as described in the literature [16, was used as substrate. Typically, the enzyme stock solution was added to a solution containing Cl-CPL (final concentrations: 50 μmol L-1 and 100 μmol L-1) and inhibitor (five different final concentrations in the range of 0.5Ki to 2Ki μmol L-1) in 0.05 mol L-1 Tris/0.5 mol L-1 NaCl, pH 7.5 buffer solution 1 mL cuvette, and the change in absorbance at 320 nm was measured immediately using a Perkin-Elmer HP 8453 UV/Vis spectrometer at 25 °
- i values were then estimated from the semireciprocal plot of the initial velocity versus the concentration of the inhibitor according to the method of Dixon [17]. The correlation coefficients for the Dixon plots were above 0.990.

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