Purification and characterization of a native lytic polysaccharide monooxygenase from Thermoascus aurantiacus

• Published in: Biotechnology letters Vol. 42; no. 10; pp. 1897 - 1905
• Main Authors: Fritsche, Susanne, Hopson, Cynthia, Gorman, Jennifer, Gabriel, Raphael, Singer, Steven W.
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Netherlands 01.10.2020; Springer Nature B.V; Springer
• Subjects: Applied Microbiology; Ascomycota; BASIC BIOLOGICAL SCIENCES; Biochemistry; Biomass deconstruction; Biomedical and Life Sciences; Biotechnology; Cell culture; Cellobiohydrolase; Cellulase; Cellulose; cellulose 1,4-beta-cellobiosidase; Colorimetry; corn stover; culture filtrates; Depolymerization; endo-1,4-beta-glucanase; Endoglucanase; Filtrate; Fungi; hydrolysis; Life Sciences; Lytic polysaccharide monooxygenase; Microbiology; Monooxygenase; Original Research Paper; oxidation; Polysaccharides; Proteins; Purification; Stover; Thermoascus aurantiacus; Biomass deconstruction; Lytic polysaccharide monooxygenase; Cellulose
• ISSN: 0141-5492; 1573-6776; 1573-6776
• DOI: 10.1007/s10529-020-02942-w

Lytic polysaccharide monooxygenases (LPMOs) have emerged as key proteins for depolymerization of cellulose. These copper-containing enzymes oxidize C-1 and/or C-4 bonds in cellulose, promoting increased hydrolysis of the oxidized cellulose chains. The LPMO from Thermoascus aurantiacus , a thermophilic ascomycete fungus, has been extensively studied and has served as a model LPMO. A method was developed to purify the LPMO from culture filtrates of T. aurantiacus along with its native cellobiohydrolase and endoglucanase. The activity of the purified LPMO was measured with a colorimetric assay that established the T opt of the native LPMO at 60 °C. Purification of the components of the T. aurantiacus cellulase mixture also enabled quantification of the amounts of cellobiohydrolase, endoglucanase and LPMO present in the T. aurantiacus culture filtrate, establishing that the LPMO was the most abundant protein in the culture supernatants. The importance of the LPMO to activity of the mixture was demonstrated by saccharifications with Avicel and acid-pretreated corn stover.