A simple enzymatic assay for the quantification of C1-specific cellulose oxidation by lytic polysaccharide monooxygenases.
KELLER, M. B; FELBY, C; LABATE, Carlos Alberto; PELLEGRINI, Vanessa de Oliveira Arnoldi; HIGASI, Paula Miwa Rabêlo; SINGH, R. K; POLIKARPOV, Igor; BLOSSOM, B. M.
KELLER, M. B; FELBY, C; LABATE, Carlos Alberto; PELLEGRINI, Vanessa de Oliveira Arnoldi; HIGASI, Paula Miwa Rabêlo; SINGH, R. K; POLIKARPOV, Igor; BLOSSOM, B. M.
Abstract: Objective The development of an enzymatic assay for the specific quantification of the C1-oxidation product, i.e. gluconic acid of cellulose active lytic polysaccharide monooxygenases (LPMOs). Results In combination with a b-glucosidase, the spectrophotometrical assay can reliably quantify the specific C1- oxidation product of LPMOs acting on cellulose. It is applicable for a pure cellulose model substrate as well as lignocellulosic biomass. The enzymatic assay compares well with the quantification performed by HPAEC-PAD. In addition, we show that simple boiling is not sufficient to inactivate LPMOs and we suggest to apply a metal chelator in addition to boiling or to drastically increase pH for proper inactivation. Conclusions We conclude that the versatility of this simple enzymatic assay makes it useful in a wide range of experiments in basic and applied LPMO research and without the need for expensive instrumentation, e.g. HPAEC-PAD.
@article={002987301,author = {KELLER, M. B; FELBY, C; LABATE, Carlos Alberto; PELLEGRINI, Vanessa de Oliveira Arnoldi; HIGASI, Paula Miwa Rabêlo; SINGH, R. K; POLIKARPOV, Igor; BLOSSOM, B. M.},title={A simple enzymatic assay for the quantification of C1-specific cellulose oxidation by lytic polysaccharide monooxygenases},journal={Biotechnology Letters},note={v. 42, n. 1, p. 93-102},year={2020}}