Purification and Structural Characterization of the Auxiliary Activity 9 Native Lytic Polysaccharide Monooxygenase from <i>Thermoascus aurantiacus</i> and Identification of Its C1- and C4-Oxidized Reaction Products
Auxiliary activity 9 (AA9) lytic polysaccharide monooxygenases (LPMOs) are copper-dependent oxidoreductases that use O<sub>2</sub> or H<sub>2</sub>O<sub>2</sub> to perform oxidative cleavage of cellulose in the presence of an electron donor. Combined with cellulas...
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2022-01-01
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| author | Weishuai Yu Imran Mohsin Anastassios C. Papageorgiou Duochuan Li |
| author_facet | Weishuai Yu Imran Mohsin Anastassios C. Papageorgiou Duochuan Li |
| author_sort | Weishuai Yu |
| collection | DOAJ |
| description | Auxiliary activity 9 (AA9) lytic polysaccharide monooxygenases (LPMOs) are copper-dependent oxidoreductases that use O<sub>2</sub> or H<sub>2</sub>O<sub>2</sub> to perform oxidative cleavage of cellulose in the presence of an electron donor. Combined with cellulases, they can assist in a more efficient cleavage of cellulose. AA9 LPMOs have therefore attracted considerable attention in recent years for use in biotechnological applications. Here, a native AA9 LPMO (n<i>Ta</i>AA9A) from the thermophilic fungus <i>Thermoascus aurantiacus</i> was purified and characterized. The enzyme was shown to be active and able to cleave cellulose and xylan to produce C1- and C4-oxidized products. It was also found to retain about 84.3, 63.7, and 35.3% of its activity after incubation for 30 min at 60, 70, and 80 °C, respectively, using quantitative activity determination. The structure was determined to 1.36 Å resolution and compared with that of the recombinant enzyme expressed in <i>Aspergillus oryzae</i>. Structural differences in the glycosylated Asn138 and in solvent-exposed loops were identified. |
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| issn | 2073-4344 |
| language | English |
| last_indexed | 2024-03-09T22:22:29Z |
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| spelling | doaj.art-b3b840dcea9443c1ab0f974f1063c5e72023-11-23T19:11:53ZengMDPI AGCatalysts2073-43442022-01-0112213910.3390/catal12020139Purification and Structural Characterization of the Auxiliary Activity 9 Native Lytic Polysaccharide Monooxygenase from <i>Thermoascus aurantiacus</i> and Identification of Its C1- and C4-Oxidized Reaction ProductsWeishuai Yu0Imran Mohsin1Anastassios C. Papageorgiou2Duochuan Li3Department of Mycology, Shandong Agricultural University, Taian 271018, ChinaTurku Bioscience Centre, University of Turku, 20521 Turku, FinlandTurku Bioscience Centre, University of Turku, 20521 Turku, FinlandDepartment of Mycology, Shandong Agricultural University, Taian 271018, ChinaAuxiliary activity 9 (AA9) lytic polysaccharide monooxygenases (LPMOs) are copper-dependent oxidoreductases that use O<sub>2</sub> or H<sub>2</sub>O<sub>2</sub> to perform oxidative cleavage of cellulose in the presence of an electron donor. Combined with cellulases, they can assist in a more efficient cleavage of cellulose. AA9 LPMOs have therefore attracted considerable attention in recent years for use in biotechnological applications. Here, a native AA9 LPMO (n<i>Ta</i>AA9A) from the thermophilic fungus <i>Thermoascus aurantiacus</i> was purified and characterized. The enzyme was shown to be active and able to cleave cellulose and xylan to produce C1- and C4-oxidized products. It was also found to retain about 84.3, 63.7, and 35.3% of its activity after incubation for 30 min at 60, 70, and 80 °C, respectively, using quantitative activity determination. The structure was determined to 1.36 Å resolution and compared with that of the recombinant enzyme expressed in <i>Aspergillus oryzae</i>. Structural differences in the glycosylated Asn138 and in solvent-exposed loops were identified.https://www.mdpi.com/2073-4344/12/2/139monooxygenaseoxidoreductasecoppercellulose degradationthermostabilitythermophilic fungus |
| spellingShingle | Weishuai Yu Imran Mohsin Anastassios C. Papageorgiou Duochuan Li Purification and Structural Characterization of the Auxiliary Activity 9 Native Lytic Polysaccharide Monooxygenase from <i>Thermoascus aurantiacus</i> and Identification of Its C1- and C4-Oxidized Reaction Products Catalysts monooxygenase oxidoreductase copper cellulose degradation thermostability thermophilic fungus |
| title | Purification and Structural Characterization of the Auxiliary Activity 9 Native Lytic Polysaccharide Monooxygenase from <i>Thermoascus aurantiacus</i> and Identification of Its C1- and C4-Oxidized Reaction Products |
| title_full | Purification and Structural Characterization of the Auxiliary Activity 9 Native Lytic Polysaccharide Monooxygenase from <i>Thermoascus aurantiacus</i> and Identification of Its C1- and C4-Oxidized Reaction Products |
| title_fullStr | Purification and Structural Characterization of the Auxiliary Activity 9 Native Lytic Polysaccharide Monooxygenase from <i>Thermoascus aurantiacus</i> and Identification of Its C1- and C4-Oxidized Reaction Products |
| title_full_unstemmed | Purification and Structural Characterization of the Auxiliary Activity 9 Native Lytic Polysaccharide Monooxygenase from <i>Thermoascus aurantiacus</i> and Identification of Its C1- and C4-Oxidized Reaction Products |
| title_short | Purification and Structural Characterization of the Auxiliary Activity 9 Native Lytic Polysaccharide Monooxygenase from <i>Thermoascus aurantiacus</i> and Identification of Its C1- and C4-Oxidized Reaction Products |
| title_sort | purification and structural characterization of the auxiliary activity 9 native lytic polysaccharide monooxygenase from i thermoascus aurantiacus i and identification of its c1 and c4 oxidized reaction products |
| topic | monooxygenase oxidoreductase copper cellulose degradation thermostability thermophilic fungus |
| url | https://www.mdpi.com/2073-4344/12/2/139 |
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