Abstract − Analytical Sciences, 32(8), 901 (2016).
Colorimetric Coupled Enzyme Assay for Cystathionine β-Synthase
Marco ROCCHICCIOLI, Roberta MOSCHINI, Laura CAPPIELLO, Francesco BALESTRI, Mario CAPPIELLO, Umberto MURA, and Antonella DEL-CORSO
University of Pisa, Department of Biology, Biochemistry Unit, via S. Zeno, 51, 56123, Pisa, Italy
A colorimetric coupled enzyme assay for the determination of cystathionine β-synthase activity is described. The method exploits cystathionine γ-lyase as an ancillary enzyme capable of transforming cystathionine, produced by cystathionine β-synthase, into cysteine. The cysteine is then spectrophotometrically detected at 560 nm, after its specific complexation with ninhydrin. This method was used to detect cystathionine β-synthase in crude extracts, and for the kinetic characterization of the enzyme partially purified from bovine kidney. A rapid two-step protocol is described for the partial purification of cystathionine γ-lyase from bovine kidney, aimed at a suitable and stable ancillary enzyme preparation.
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