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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

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

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Author(s):
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]
Total Authors: 8
Affiliation:
[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
Total Affiliations: 4
Document type: Journal article
Source: Biotechnology Letters; v. 42, n. 1, p. 93-102, JAN 2020.
Web of Science Citations: 8
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 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)

FAPESP's process: 17/01165-4 - Enzymes and electrons at work: dual processing of carbohydrates and lignin producing prebiotic oligosaccharides and bioactive lignins
Grantee:Carlos Alberto Labate
Support Opportunities: Regular Research Grants
FAPESP's process: 15/13684-0 - Structural and functional studies of enzymes that participate in complex carbohydrates synthesis and degradation
Grantee:Igor Polikarpov
Support Opportunities: Research Projects - Thematic Grants