A simple enzymatic assay for the quantification of C1-specific cellulose oxidation by lytic polysaccharide monooxygenases

Autor(es)

Keller, M. B. ^[1] ; Felby, C. ^[1] ; Labate, C. A. ^[2] ; Pellegrini, V. O. A. ^[3] ; Higasi, P. ^[3] ; Singh, R. K. ^[4] ; Polikarpov, I. ^[3] ; Blossom, B. M. ^[1]

Afiliação do(s) autor(es)

^[1] Univ Copenhagen, Dept Geosci & Nat Resource Management, Copenhagen - Denmark

^[2] Univ Sao Paulo, Luiz Queiroz Coll Agr, Piracicaba - Brazil

^[3] Univ Sao Paulo, Sao Carlos Inst Phys, Sao Carlos - Brazil

^[4] Univ Copenhagen, Dept Chem, Copenhagen - Denmark

• Tipo de documento: Artigo Científico
• Fonte: Biotechnology Letters; v. 42, n. 1, p. 93-102, JAN 2020.
• Citações Web of Science: 0

Resumo

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 beta-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. (AU)

• Processo FAPESP: 15/13684-0 - Estudos estruturais e funcionais de enzimas que participam na síntese e degradação de carboidratos complexos
• Beneficiário: Igor Polikarpov
• Linha de fomento: Auxílio à Pesquisa - Temático
• Processo FAPESP: 17/01165-4 - Enzimas e eletrons ao trabalho: duplo processamento de carboidratos e lignina produzindo oligossacarídeos probióticos e ligninas bioativas
• Beneficiário: Carlos Alberto Labate
• Linha de fomento: Auxílio à Pesquisa - Regular