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Characterization of a Dual Cellulolytic/Xylanolytic AA9 Lytic Polysaccharide Monooxygenase from Thermothelomyces thermophilus and its Utilization Toward Nanocellulose Production in a Multi-Step Bioprocess
ACS Sustainable Chemistry & Engineering ( IF 8.4 ) Pub Date : 2022-06-24 , DOI: 10.1021/acssuschemeng.2c02255 Koar Chorozian 1 , Anthi Karnaouri 1, 2 , Antonis Karantonis 2 , Maria Souli 3 , Evangelos Topakas 1
ACS Sustainable Chemistry & Engineering ( IF 8.4 ) Pub Date : 2022-06-24 , DOI: 10.1021/acssuschemeng.2c02255 Koar Chorozian 1 , Anthi Karnaouri 1, 2 , Antonis Karantonis 2 , Maria Souli 3 , Evangelos Topakas 1
Affiliation
Lytic polysaccharide monooxygenases (LPMOs) are enzymes able to act on a great variety of polysaccharides by an oxidative cleavage mechanism, targeting not only crystalline substrates, such as cellulose and chitin, but also other amorphous structures including xylan, mannan, and pectin. The key role of LPMOs toward the isolation of nanocellulose from natural substrates has been demonstrated since these enzymes promote amorphogenesis of the substrate and facilitate the defibrillation process. In the present study, an AA9 LPMO from the thermophilic fungus Thermothelomyces thermophilus (TtLPMO9G) with C1-regioselectivity and a dual cellulolytic/xylanolytic activity was heterologously produced in Pichia pastoris and biochemically characterized. The enzyme was employed both as a pre- and a post-treatment step alongside with commercially available and in-house produced tailored cocktails of hemicellulases and cellulases in four-step multi-enzymatic processes for the isolation of nanoscale cellulose from OxiOrganosolv pretreated beechwood. Nanostructures obtained from each of these green bio-processes were examined for their morphological features and dimensions, crystallinity, colloidal stability, and the presence of carboxylate groups. The results demonstrate the formation of well-dispersed nanoscale cellulose in the complete absence of any chemical or mechanical treatment step and verify the importance of efficient hemicellulose removal for the isolation of nanocellulose.
中文翻译:
来自嗜热嗜热丝菌的双纤维素分解/木聚糖分解 AA9 裂解多糖单加氧酶的表征及其在多步生物过程中用于纳米纤维素生产的应用
裂解多糖单加氧酶 (LPMO) 是一种能够通过氧化裂解机制作用于多种多糖的酶,不仅靶向结晶底物,例如纤维素和几丁质,还靶向其他无定形结构,包括木聚糖、甘露聚糖和果胶。LPMOs 对从天然基质中分离纳米纤维素的关键作用已得到证实,因为这些酶促进基质的无形态发生并促进除颤过程。在本研究中,来自嗜热真菌Thermothelomyces thermophilus ( Tt LPMO9G) 的 AA9 LPMO 在毕赤酵母中异源产生,具有 C1 区域选择性和双重纤维素分解/木聚糖分解活性和生化特征。该酶被用作预处理和后处理步骤,与市售和内部生产的定制半纤维素酶和纤维素酶混合物一起在四步多酶法中用于从 OxiOrganosolv 预处理的山毛榉木中分离纳米级纤维素。检查了从这些绿色生物过程中的每一个获得的纳米结构的形态特征和尺寸、结晶度、胶体稳定性和羧酸盐基团的存在。结果表明在完全没有任何化学或机械处理步骤的情况下形成了良好分散的纳米级纤维素,并验证了有效去除半纤维素对于分离纳米纤维素的重要性。
更新日期:2022-06-24
中文翻译:
来自嗜热嗜热丝菌的双纤维素分解/木聚糖分解 AA9 裂解多糖单加氧酶的表征及其在多步生物过程中用于纳米纤维素生产的应用
裂解多糖单加氧酶 (LPMO) 是一种能够通过氧化裂解机制作用于多种多糖的酶,不仅靶向结晶底物,例如纤维素和几丁质,还靶向其他无定形结构,包括木聚糖、甘露聚糖和果胶。LPMOs 对从天然基质中分离纳米纤维素的关键作用已得到证实,因为这些酶促进基质的无形态发生并促进除颤过程。在本研究中,来自嗜热真菌Thermothelomyces thermophilus ( Tt LPMO9G) 的 AA9 LPMO 在毕赤酵母中异源产生,具有 C1 区域选择性和双重纤维素分解/木聚糖分解活性和生化特征。该酶被用作预处理和后处理步骤,与市售和内部生产的定制半纤维素酶和纤维素酶混合物一起在四步多酶法中用于从 OxiOrganosolv 预处理的山毛榉木中分离纳米级纤维素。检查了从这些绿色生物过程中的每一个获得的纳米结构的形态特征和尺寸、结晶度、胶体稳定性和羧酸盐基团的存在。结果表明在完全没有任何化学或机械处理步骤的情况下形成了良好分散的纳米级纤维素,并验证了有效去除半纤维素对于分离纳米纤维素的重要性。