在糖化用糖苷水解酶糖化前，用溶细胞多糖单加氧酶对木质纤维素进行预处理是一种节约第二代乙醇工艺的创新方法。
An innovative approach of priming lignocellulosics with lytic polysaccharide mono-oxygenases prior to saccharification with glycosyl hydrolases can economize second generation ethanol process.
机构信息
Department of Microbiology, Guru Nanak Dev University, Amritsar, Punjab 143005, India.
Department of Microbiology, Guru Nanak Dev University, Amritsar, Punjab 143005, India.
出版信息
Bioresour Technol. 2020 Jul;308:123257. doi: 10.1016/j.biortech.2020.123257. Epub 2020 Mar 26.
Two Lytic polysaccharide Mono-Oxygenases (LPMOs), non-modular (PMO_08942) and modular (PMO_07920), from thermotolerant fungus Aspergillus terreus 9DR cloned and expressed in Pichia pastoris X33 and purified to homogeneity using ion-exchange chromatography were found to be of 29 and ~40 kDa, respectively. Both LPMOs were optimally active at 50 °C; PMO_08942 was active under acidic condition (pH 5.0) and PMO_07920 at pH 7.0. Modular LPMO (PMO_07920) tethered to CBM-1 was found to be versatile as it showed appreciable activity on complex polysaccharide (both cellulose and xylans) as compared to non-modular (PMO_08942). The t of PMO_08942 (192 h, pH 5.0) and PMO_0792 (~192 h, pH 7.0) at 50 °C, suggests highly stable nature of these LPMOs. Fluorescently tagged modular AA9 was studied microscopically to understand interaction with pretreated biomass. Priming of biomass for up to 6 h with LPMOs prior to initiating hydrolysis with core cellulase enzyme resulted in significantly higher saccharification.
两种溶细胞多糖单加氧酶（LPMOs），非模块（PMO_08942）和模块（PMO_07920），来自耐热真菌 Aspergillus terreus 9DR，在巴斯德毕赤酵母 X33 中克隆和表达，并通过离子交换色谱法纯化至均一性。这两种 LPMO 的分子量分别约为 29 和 40 kDa。两种 LPMO 的最适活性温度均为 50°C；PMO_08942 在酸性条件（pH 5.0）下具有活性，而 PMO_07920 在 pH 7.0 下具有活性。与非模块（PMO_08942）相比，连接到 CBM-1 的模块 LPMO（PMO_07920）具有多功能性，因为它对复杂多糖（纤维素和木聚糖）表现出相当大的活性。PMO_08942（t = 192 h，pH 5.0）和 PMO_0792（t = 192 h，pH 7.0）在 50°C 下的半衰期表明这些 LPMO 具有高度稳定的性质。对荧光标记的模块 AA9 进行了显微镜研究，以了解其与预处理生物质的相互作用。在核心纤维素酶水解之前，用 LPMO 对生物质进行长达 6 小时的预处理，可显著提高糖化率。
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