Protein Engineering of Subtilisin

  • Thomas P. Graycar
Part of the Topics in Applied Chemistry book series (TAPP)


The use of enzymes as catalysts in organic synthesis reactions has expanded rapidly in recent years.1–5 In contrast to conventional organic reactions enzyme-catalyzed reactions offer the potential of highly stereo-selective or regioselective transformations. The value of enzymes in organic synthesis can be further attributed to the high reactivity and mild conditions characteristic of enzyme-catalyzed reactions. The increased availability and lower cost of enzymes resulting from the development of recombinant DNA technology has also stimulated interest in the use of enzymes for the production of fine organic chemicals. An apparent impediment to the widespread utilization of enzymes as commercial biocatalysts, however, is the difficulty encountered in optimizing their use in industrial processes. Although enzymes are capable of reacting with compounds having structures similar to that of their natural substrates, the catalytic efficiency of reactions involving nonnatural substrates is often suboptimal. In addition, the inherent instability of proteins and the sensitivity of enzyme activity to alterations in pH further limit their utility in industry. Hence, despite the known advantages of enzymes their use as biocatalysts is currently limited to less than 5% of the total industrial enzyme market.6 In the past, the only methods available to optimize the performance of an enzyme involved chemical modification of residues on the surface of the enzyme or else random mutagenesis of the corresponding gene. Now, however, the recent development of protein engineering has made it possible to redesign the structure of an enzyme and tailor its functional properties for a particular application, thereby greatly enhancing the potential to create novel industrial biocatalysts.

Table 1

Functional Properties of Enzymes which Can Be Modified by Protein Engineering








Catalytic rate

Organic solvents

pH Activity profile


Allosteric regulation




Catalytic Efficiency Protein Engineering Mutant Enzyme Random Mutagenesis Hybrid Protein 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


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Copyright information

© Springer Science+Business Media New York 1991

Authors and Affiliations

  • Thomas P. Graycar
    • 1
  1. 1.Department of EnzymologyGenencor International, IncSouth San FranciscoUSA

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