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Recently, we showed that Overhauser\u2010enhanced MRI is suitable to such a purpose. A \u03b2\u2010phosphorylated nitroxide substrate prototype exhibiting keto\u2013enol equilibrium upon enzymatic activity has been prepared. Upon enzymatic hydrolysis, a large variation of the phosphorus hyperfine coupling constant (\u0394<jats:italic>a<\/jats:italic><jats:sub>P<\/jats:sub>=4\u2005G) was observed. The enzymatic activities of several enzymes were conveniently monitored by electronic paramagnetic resonance (EPR). Using a 0.2\u2005T MRI machine, in\u2005vitro and in\u2005vivo OMRI experiments were successfully performed, affording a 1200\u2009% enhanced MRI signal in\u2005vitro, and a 600\u2009% enhanced signal in\u2005vivo. 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