Investigation of the Effect of Formaldehyde on the Structure of Formolase
To investigate the effect of formaldehyde on formolase and the cause of its inactivation,in our present study for-molase was incubated with different concentrations of formaldehyde(300 mmol/L,1 mol/L)for different time(1 hour,2 hours and 3 hours).Subsequently,the residual activity of formolase was detected and the modified and cross-linked sites of formolase were detected by liquid chromatography-tandem mass spectrometry(LC-MS/MS).The results showed that there was a significant cross-linking and modification phenomenon of formolase after incubation with formaldehyde,and the higher of the formaldehyde concentration and the longer the incubation time,the higher degree of cross-linking of formolase.Two formaldehyde modification sites(N186 and K228)of formolase and potential formaldehyde cross-linked fragments were identified with LC-MS/MS.Through the cross-linking reaction with such amino acids as asparagine and lysine,formaldehyde destroyed its local structure and changed the overall aggregation state,thus making it inactive.This finding might be also beneficial for providing semi-rational design sites for improving formaldehyde tolerance of formolase.
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