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Review
. 2015:59:1-41.
doi: 10.1042/bse0590001.

Enzymes: principles and biotechnological applications

Peter K Robinson  1
Affiliations
Review

Enzymes: principles and biotechnological applications

Peter K Robinson. Essays Biochem. 2015.

Erratum in

  • Essays Biochem. 2015;59:75

Abstract

Enzymes are biological catalysts (also known as biocatalysts) that speed up biochemical reactions in living organisms, and which can be extracted from cells and then used to catalyse a wide range of commercially important processes. This chapter covers the basic principles of enzymology, such as classification, structure, kinetics and inhibition, and also provides an overview of industrial applications. In addition, techniques for the purification of enzymes are discussed.

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Figures

Figure 1.
Figure 1.. Representation of substrate binding to the active site of an enzyme molecule.
Figure 2.
Figure 2.. The components of a holoenzyme.
Figure 3.
Figure 3.. Effect of an enzyme on reducing the activation energy required to start a reaction where (a) is uncatalysed and (b) is enzyme-catalysed reaction.
Figure 4.
Figure 4.. Formation of product in an enzyme-catalysed reaction, plotted against time.
Figure 5.
Figure 5.. Relationship between enzyme concentration and the rate of an enzyme-catalysed reaction.
Figure 6.
Figure 6.. Relationship between substrate concentration and the rate of an enzyme-catalysed reaction.
Figure 7.
Figure 7.. (a) Direct plot. (b) Lineweaver–Burk plot of the same kinetic data.
Figure 8.
Figure 8.. Lineweaver–Burk plot of similar kinetic data, which differ only in a single. (Final data point (a) 1/v 0.03 at 1/S of 0.2 and (b) 1/v 0.031 at 1/S of 0.18).
Figure 9.
Figure 9.. The pH profile of β-glucosidase.
Figure 10.
Figure 10.. The effect of temperature on enzyme activity.
Figure 11.
Figure 11.. Activity/substrate profiles of allosteric (^) and non-allosteric (•) enzymes with the same affinity and maximal velocity.
Figure 12.
Figure 12.. Stirred-tank reactor containing immobilized enzyme.
Figure 13.
Figure 13.. Continuous-flow reactors. (a) Continuous-flow stirred-tank reactor (CSTR). (b) Packed-bed reactor (PBR).
Figure 14.
Figure 14.. Immobilization techniques.
Figure 15.
Figure 15.. Properties of (a) DEAE–Sephadex and (b) CM–Sephadex ion-exchange resins.
Figure 16.
Figure 16.. A laboratory-scale glucose analyser. Photograph supplied courtesy of YSI (UK) Limited.
Figure 17.
Figure 17.. A hand-held glucose biosensor suitable for personal use. Photograph supplied courtesy of Abbott Diabetes Care.
See this image and copyright information in PMC

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References

Recommended reading and key publications

1. Historically important landmark papers (in chronological order)
    1. Takamine J. Process of making diastatic enzyme. 1894 U.S. Pat. 525,823. Describes the first commercial exploitation of semi-purified enzymes in the West.
    1. Briggs G.E., Haldane J.B.S. A note on the kinetics of enzyme action. Biochem. J. 1925;19:338–339. A classic paper in which the steady-state assumption was introduced into the derivation of the Michaelis–Menten equation. - PMC - PubMed
    1. Koshland D.E., Jr Application of a theory of enzyme specificity to protein synthesis. Proc. Natl Acad. Sci. U.S.A. 1958;44:98–104. Describes the proposal of an ‘induced fit’ mechanism of substrate binding. - PMC - PubMed
    1. Clark L.C., Jr, Lyons C. Electrode systems for continuous monitoring in cardiovascular surgery. Ann. N.Y. Acad. Sci. 1962;102:29–45. Introduces the concept of a biosensor for measuring blood glucose levels during surgery. - PubMed
    1. Monod J., Wyman H., Changeux J.P. On the nature of allosteric transitions: a plausible model. J. Mol. Biol. 1965;12:88–118. Describes the ‘concerted’ model of transitions of allosteric proteins in which all constituent monomers are in either the T-state or the R-state. - PubMed
2. Enzyme principles
    1. Changeux J.-P. 50 years of allosteric interactions: the twists and turns of the models. Nat. Rev. Mol. Cell Biol. 2013;14:819–829. - PubMed
    1. Kamata K., Mitsuya M., Nishimura T., Eiki J., Nagata Y. Structural basis for allosteric regulation of the monomeric allosteric enzyme human glucokinase. Structure. 2004;12:429–438. - PubMed
3. Enzyme applications
    1. Adrio J.L., Demain A.L. Microbial enzymes: tools for biotechnological processes. Biomolecules. 2014;4:117–139. - PMC - PubMed
    1. Clarke S.F., Foster J.R. A history of blood glucose meters and their role in self-monitoring of diabetes mellitus. Br. J. Biomed. Sci. 2012;69:83–93. - PubMed
    1. Fernandes P. Enzymes in food processing: a condensed overview on strategies for better biocatalysts. Enzyme Res. 2010;2010:862537. - PMC - PubMed
    1. Vashist S.K., Zheng D., Al-Rubeaan K., Luong J.H.T., Sheu F.-S. Technology behind commercial devices for blood glucose monitoring in diabetes management: a review. Anal. Chim. Acta. 2011;703:124–136. - PubMed
    1. Vellard M. The enzyme as drug: application of enzymes as pharmaceuticals. Curr. Opin. Biotechnol. 2003;14:444–450. - PubMed
4. Useful textbooks
    1. Bisswanger H. Enzyme Kinetics: Principles and Methods. 2nd edn. Weinheim, Germany: Wiley-VCH; 2008. Available online and as hard copy. A user-friendly and comprehensive treatise on enzyme kinetics.
    1. Buchholz K., Kasche V., Bornscheuer U.T. Biocatalysts and Enzyme Technology. 2nd edn. Weinheim, Germany: Wiley-VCH; 2012. Best-selling textbook that provides an instructive and comprehensive overview of our current knowledge of biocatalysis and enzyme technology.
    1. Copeland R.A. In: Evaluation of Enzyme Inhibitors in Drug Discovery: A Guide for Medicinal Chemists and Pharmacologists. 2nd edn. Hoboken NJ., editor. John Wiley & Sons, Inc; 2013. Provides thorough coverage of both the principles and applications of enzyme inhibitors. - PubMed
    1. McGrath M.J., Scanaill C.N. Sensor Technologies: Healthcare, Wellness and Environmental Applications. New York: Apress Media, LLC; 2014. Available online. Covers sensor technologies and their clinical applications, together with broader applications that are relevant to wellness, fitness, lifestyle and the environment.
    1. Trevan M.D. Immobilized Enzymes: An Introduction and Applications in Biotechnology. Chichester: John Wiley & Sons; 1980. An older text, and difficult to find except in libraries, but it provides an introductory text for non-experts, and as yet there is no other book that fulfils this role.