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Title
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High-level expression, purification, and crystallization of recombinant spinach glycolate oxidase in Escherichia coli.
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Authors
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K.Stenberg,
Y.Lindqvist.
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Ref.
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Protein Expr Purif, 1996,
8,
295-298.
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PubMed id
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Abstract
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Glycolate oxidase is a flavin-dependent enzyme in the photorespiratory pathway
in plants. Here we report the heterologous expression of glycolate oxidase in
Escherichia coli and an isolation procedure which results in 4 mg pure protein
per gram cell paste in only 1.5 days. This corresponds to a more than 50-fold
improvement in yield compared to previously reported expression systems. The
purified recombinant protein can be crystallized easily, and the crystals are
isomorphous to those obtained from the protein isolated from spinach. The
availability of large amounts of enzyme will be a great advantage in the 3D
structural and biochemical studies of mutants and inhibitor complexes.
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