Abstract
Enzyme and substrate form an ES-complex, which reacts further to enzyme and product. This process requires time, so each enzyme molecule can only handle a certain number of substrate molecules per unit time, called the turnover-number k cat. This number multiplied with the number of enzyme molecules is the limiting reaction velocity, V max, reached only at infinite substrate concentration. At [S] < ∞ enzyme molecules need time to find a new substrate after the release of product, hence v < V max.
Measuring the enzyme concentration in biological fluids is of great clinical significance.
Some enzymes are produced as inactive precursors and activated only when needed.
For some enzymes, the molecular mechanism of their action has been determined. If there are several substrates and/or products, substrates can bind to, and products be released from enzymes in specific order or randomly. Reaction mechanism is determined by keeping the concentration of one substrate constant, while varying the second. Different enzyme mechanisms result in characteristic Lineweaver-Burk-plots.
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Notes
- 1.
Henri’s version of the HMM equation tries to take care of the increasing product concentration by treating the product as a competitive inhibitor of the reaction:
$$\displaystyle{ v = \frac{V _{\mathrm{max}}([S]_{0} - [P])} {1 + \frac{[S]_{0}-[P]} {K_{\mathrm{s}}} + \frac{[P]} {K_{p}}} }$$(5.28)with K s and K p the dissociation constants for substrate and product, respectively. [S]0 is the substrate concentration at the start of the experiment.
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Buxbaum, E. (2015). Enzyme Kinetics and Mechanism. In: Fundamentals of Protein Structure and Function. Springer, Cham. https://doi.org/10.1007/978-3-319-19920-7_5
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