Acetylacetone extends the working life of laccase in enzymatic transformation of malachite green by interfering with a key intermediate.

Abstract

The potential of acetylacetone (AA) as a mediator of laccase has been tested in the enzymatic transformation of malachite green (MG). AA inhibited the laccase-induced transformation of MG at the beginning of incubation but extended the working life of laccase in long runs. To elucidate the underlying mechanisms, the transformation of MG in the laccase-AA system was systematically investigated. The inhibition of AA on the enzymatic transformation of MG conformed to the partial mixed model. The transformation of N,N,N',N'-tetramethyl-1,1'-biphenyl-4,4'-diamine (NTB) was identified as the rate-controlling step in the laccase system. The generated NTB was oxidized to NTB+ by laccase, which acted as a redox mediator to accelerate the transformation of MG. The addition of AA to the enzymatic system quenched the NTB+ by forming an intermediate complex of AA-NTB. This quenching reaction led to two contrary effects: the acceleration caused by NTB+ in the enzymatic transformation of MG was inhibited whereas the formation of AA-NTB complex enhanced the further transformation at the later stage. As a result, less laccase was consumed, which explained the extended working life of laccase in the long runs. The understanding of these mechanisms are helpful for the better use of laccase as a green biocatalyst.