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- W2011307452 abstract "The gene encoding fructokinase (EC 2.7.1.4) fromZymomonas mobilishas been expressed at high level inEscherichia coliby modifying the ribosome binding site using the polymerase chain reaction. A simple two-step purification from extracts of the recombinant cells results in highly purified enzyme suitable for use in fructose determination. Using the polymerase chain reaction in mutagenic conditions, a variant of fructokinase was isolated which was more thermostable than the wild type, taking the 30 min half-life from 70.1 to 72.4°C. The purified thermostable variant had the same specific activity as the wild type. Sequencing of the variant indicated that only one amino acid was changed, with Ser 69 becoming Ala. Searches of the mutant libraries for variants that were (a) active with glucose or (b) had reduced inhibition by glucose were unsuccessful." @default.
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- W2011307452 date "1996-06-01" @default.
- W2011307452 modified "2023-10-17" @default.
- W2011307452 title "Overexpression, Purification, and Generation of a Thermostable Variant ofZymomonas mobilisFructokinase" @default.
- W2011307452 doi "https://doi.org/10.1006/prep.1996.0055" @default.
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