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Promoting and Impeding Effects of Lytic Polysaccharide Monooxygenases on Glycoside Hydrolase Activity
ACS Sustainable Chemistry & Engineering ( IF 7.1 ) Pub Date : 2020-08-25 , DOI: 10.1021/acssuschemeng.0c04779 Malene Billeskov Keller 1 , Silke Flindt Badino 2 , Benedikt Möllers Blossom 1 , Brett McBrayer 3 , Kim Borch 4 , Peter Westh 2
ACS Sustainable Chemistry & Engineering ( IF 7.1 ) Pub Date : 2020-08-25 , DOI: 10.1021/acssuschemeng.0c04779 Malene Billeskov Keller 1 , Silke Flindt Badino 2 , Benedikt Möllers Blossom 1 , Brett McBrayer 3 , Kim Borch 4 , Peter Westh 2
Affiliation
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Lytic polysaccharide monooxygenases (LPMOs) have attracted attention due to their ability to boost cellulolytic enzyme cocktails for application in biorefineries. However, the interplay between LPMOs and individual glycoside hydrolases remains poorly understood. We investigated how the activity of two cellobiohydrolases (Cel7A and Cel6A) and an endoglucanase (Cel7B) from Trichoderma reesei were affected by a C1-oxidizing LPMO from Thielavia terrestris (TtAA9). We quantified products from a mixture of LPMO and glycoside hydrolase and estimated separate contributions of products by each of the enzymes. Hereby, we assessed if an observed synergy reflected a promotion of the activity of hydrolase, LPMO, or both. We consistently found that TtAA9 affected the investigated hydrolases differently. It strongly impeded the turnover of the reducing end cellobiohydrolase, TrCel7A, moderately promoted the turnover of the nonreducing end cellobiohydrolase TrCel6A, and promoted the turnover of the endoglucanase, TrCel7B up to 5-fold. The promoting effect on the endoglucanase increased with hydrolysis extent, indicating that the promoting effect became more important as the recalcitrance of the substrate increased. Experiments with mixtures containing multiple glycoside hydrolases suggested that the LPMO primarily promoted the activity of the endoglucanase, whereas promotion of TrCel6A was secondary.
中文翻译：
溶胞多糖单加氧酶对糖苷水解酶活性的促进和阻碍作用
溶菌多糖单加氧酶（LPMO）由于其增强纤维素分解酶混合物的能力而备受关注，可用于生物精炼厂。但是，LPMOs和各个糖苷水解酶之间的相互作用仍然知之甚少。我们调查了来自里氏木霉的两个纤维二糖水解酶（Cel7A和Cel6A）和一种内切葡聚糖酶（Cel7B）的活性如何受到土生thielavia terrestris（Tt AA9）的C1氧化LPMO的影响。我们从LPMO和糖苷水解酶的混合物中定量了产物，并估计了每种酶对产物的独立贡献。因此，我们评估了观察到的协同作用是否反映了水解酶，LPMO或两者的活性的提高。我们一贯发现，Tt的AA9对研究的水解酶的影响不同。它强烈地阻碍了还原末端纤维二糖水解酶Tr Cel7A的营业额，适度地促进了非还原末端纤维二糖水解酶Tr Cel6A的营业额，并且将内切葡聚糖酶Tr Cel7B的营业额提高了5倍。随着水解程度的增加，对内切葡聚糖酶的促进作用增加，表明随着底物的抗拒性增加，促进作用变得更加重要。含有多种糖苷水解酶的混合物的实验表明，LPMO主要促进内切葡聚糖酶的活性，而Tr Cel6A的促进则是次要的。
更新日期：2020-09-21
中文翻译：
溶胞多糖单加氧酶对糖苷水解酶活性的促进和阻碍作用
溶菌多糖单加氧酶（LPMO）由于其增强纤维素分解酶混合物的能力而备受关注，可用于生物精炼厂。但是，LPMOs和各个糖苷水解酶之间的相互作用仍然知之甚少。我们调查了来自里氏木霉的两个纤维二糖水解酶（Cel7A和Cel6A）和一种内切葡聚糖酶（Cel7B）的活性如何受到土生thielavia terrestris（Tt AA9）的C1氧化LPMO的影响。我们从LPMO和糖苷水解酶的混合物中定量了产物，并估计了每种酶对产物的独立贡献。因此，我们评估了观察到的协同作用是否反映了水解酶，LPMO或两者的活性的提高。我们一贯发现，Tt的AA9对研究的水解酶的影响不同。它强烈地阻碍了还原末端纤维二糖水解酶Tr Cel7A的营业额，适度地促进了非还原末端纤维二糖水解酶Tr Cel6A的营业额，并且将内切葡聚糖酶Tr Cel7B的营业额提高了5倍。随着水解程度的增加，对内切葡聚糖酶的促进作用增加，表明随着底物的抗拒性增加，促进作用变得更加重要。含有多种糖苷水解酶的混合物的实验表明，LPMO主要促进内切葡聚糖酶的活性，而Tr Cel6A的促进则是次要的。
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