A steady state theory for processive cellulases

Nicolaj Cruys-Bagger, Jens Elmerdahl Olsen, Eigil Præstgaard, Kim Borch, Peter Westh

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    Abstract

    Processive enzymes perform sequential steps of catalysis without dissociating from their polymeric substrate. This mechanism is considered essential for efficient enzymatic hydrolysis of insoluble cellulose (particularly crystalline cellulose), but a theoretical framework for processive kinetics remains to be fully developed. In this paper, we suggest a deterministic kinetic model that relies on a processive set of enzyme reactions and a quasi steady-state assumption. It is shown that this approach is practicable in the sense that it leads to mathematically simple expressions for the steady-state rate, and only requires data from standard assay techniques as experimental input. Specifically, it is shown that the processive reaction rate at steady state may be expressed by a hyperbolic function related to the conventional Michaelis–Menten equation. The main difference is a ‘kinetic processivity coefficient’, which represents the probability of the enzyme dissociating from the substrate strand before completing n sequential catalytic steps, where n is the mean processivity number measured experimentally. Typical processive cellulases have high substrate affinity, and therefore this probability is low. This has significant kinetic implications, for example the maximal specific rate (Vmax/E0) for processive cellulases is much lower than the catalytic rate constant (kcat). We discuss how relationships based on this theory may be used in both comparative and mechanistic analyses of cellulases.
    Original languageEnglish
    JournalF E B S Journal
    Volume280
    Issue number16
    Pages (from-to)3952-3961
    Number of pages9
    ISSN1742-464X
    DOIs
    Publication statusPublished - 2013

    Cite this

    Cruys-Bagger, N., Olsen, J. E., Præstgaard, E., Borch, K., & Westh, P. (2013). A steady state theory for processive cellulases. F E B S Journal, 280(16), 3952-3961. https://doi.org/10.1111/febs.12397