A continuous-flow system for high-precision kinetics using small volumes

Xianzhi Zhou, Rohit Medhekar, Michael D. Toney

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

Abstract

A generally applicable continuous-flow kinetic analysis system that gives data of a precision high enough to measure small kinetic isotope effects for enzymatic and nonenzymatic reactions is described. It employs commercially available components that are readily assembled into an apparatus that is easy to use. It operates under laminar flow conditions, which requires that the time between the initiation of the reaction in the mixer and the observation be long enough that molecular diffusion can effect a symmetrization of the concentration profile that results from a thin plug of reagents introduced at the mixer. The analysis of a second-order irreversible reaction under pseudo-first-order conditions is presented. The Yersinia pestis protein tyrosine phosphatase catalyzed hydrolysis of p-nitrophenyl phosphate is characterized with the system, and a proton inventory on kcat is presented.

Original languageEnglish (US)
Pages (from-to)3681-3687
Number of pages7
JournalAnalytical Chemistry
Volume75
Issue number15
DOIs
StatePublished - Aug 1 2003

ASJC Scopus subject areas

  • Analytical Chemistry

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