Crude enzyme reaction: A total of 1 mL system containing 12 mM p-nitrobenzaldehyde, 48 mM nitromethane, crude enzyme and solvent water, was reacted at 30 °C in a metal bath for 10 min. 10 μL of the reaction solution was taken and diluted 50 times with isopropanol and filtered through 0.25 μm organic filter membranes. HPLC was used for analysis.
Pure enzyme reaction: A total of 1 mL system containing 12 mM p-nitrobenzaldehyde, 48 mM nitromethane, 0.3 mg/mL pure enzyme and solvent water, was reacted at 30 °C in a metal bath for 10 min. 10 μL of the reaction solution was taken and diluted 50 times with isopropanol and filtered through 0.25 μm organic filter membranes. HPLC was used for analysis, and the yield was calculated using formula: Yield(\% )=product/substrateint×100%
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