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Directed evolution of yeast pyruvate decarboxylase 1 for attenuated regulation and increased stability

Stevenson, Bradley; Liu, Jian-Wei; Ollis, David


Five generations of directed evolution resulted in yeast pyruvate decarboxylase 1 (Pdc1) variants with improved activity for 1 mM pyruvate at pH 7.5 in the presence of phosphate. The best variant, named 5LS30, contained the following mutations: A143T, T156A, Q367H, N396I, and K478R. In comparison with native Pdc1, 5LS30 had the substrate concentration required for half-saturation reduced by almost 3-fold at pH 7.5 and the phosphate inhibition reduced by 4-fold at pH 6.0. The apparent...[Show more]

CollectionsANU Research Publications
Date published: 2008
Type: Journal article
Source: Biochemistry
DOI: 10.1021/bi701858u


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