Bio/Technology
9, 738 - 742 (1991)
doi:10.1038/nbt0891-738
Enhancing the Thermostability of Glucose Isomerase by Protein EngineeringWim J. Quax1, *, Nadir T. Mrabet2, *, ‡, Ruud G. M. Luiten1, Paul W. Schuurhuizen1, Patrick Stanssens2
& Ignace Lasters2
1Gist-brocades, Research & Development, Wateringseweg 1, 2611 XT Delft, The Netherlands.
2Plant Genetic Systems, Protein Engineering Dept., 22 Jozef Plateau Straat, B-9000, Gent, Belgium.
*Corresponding authors.
‡Present address: Université Libre de Bruxelles, Unité Conformation des Macromolécules Biologiques, Avenue Paul Heger, B1050. Brussels, Belgium. We have engineered recombinant glucose isomerase (GI) from Actinoplanes missouriensis by site-directed mutagenesis to enhance its thermal stability in both the soluble and immobilized forms. Substitution of arginine for lysine at position 253, which lies at the dimer/dimer interface of the GI tetramer, produced the largest stabilization under model industrial conditions. We discuss our results in terms of a model in which chemical glycation of lysines by sugars in the industrial corn syrup substrate represents a major pathway of destabilization.
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