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A tryptophan-shift variant of Bacillus stearothermophilus phosphofructokinase (BsPFK), W179F/F76W, was constructed to evaluate the binding and allosteric characteristics associated with MgATP. W179F/F76W BsPFK has a specific activity of 77±1U/mg at pH 7 and 25°C, which is a 35% decrease compared to the wild-type enzyme. The K m for MgATP increases from 43±3μM for wild-type BsPFK to 160±7μM...
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