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

Fritsche, Susanne; Hopson, Cynthia; Gorman, Jennifer; Gabriel, Raphael; Singer, Steven W.

doi:10.1007/s10529-020-02942-w

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

Journal Article · Mon Jun 15 00:00:00 EDT 2020 · Biotechnology Letters

DOI:https://doi.org/10.1007/s10529-020-02942-w· OSTI ID:1844511

Fritsche, Susanne ^[1]; Hopson, Cynthia ^[2]; Gorman, Jennifer ^[3]; Gabriel, Raphael ^[4]; ^[3]

Lawrence Berkeley National Lab. (LBNL), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Univ. of Natural Resources and Life Sciences Vienna (Austria)
Lawrence Berkeley National Lab. (LBNL), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Complutense Univ. of Madrid (Spain)
Lawrence Berkeley National Lab. (LBNL), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Lawrence Berkeley National Lab. (LBNL), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Technische Univ. Braunschweig (Germany)

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. Here, the importance of the LPMO to activity of the mixture was demonstrated by saccharifications with Avicel and acid-pretreated corn stover.

View Accepted Manuscript (DOE)

Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 1844511

Journal Information:: Biotechnology Letters, Journal Name: Biotechnology Letters Journal Issue: 10 Vol. 42; ISSN 0141-5492

Publisher:: SpringerCopyright Statement

Country of Publication:: United States

Language:: English

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