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首页> 外文期刊>Applied Microbiology >Activating Intrinsic Carbohydrate-Active Enzymes of the Smut Fungus Ustilago maydis for the Degradation of Plant Cell Wall Components
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Activating Intrinsic Carbohydrate-Active Enzymes of the Smut Fungus Ustilago maydis for the Degradation of Plant Cell Wall Components

机译：激活Smut Fungus ustilago maydis的内在碳水化合物活性酶用于植物细胞壁部件的降解

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The microbial conversion of plant biomass to valuable products in a consolidated bioprocess could greatly increase the ecologic and economic impact of a biorefinery. Current strategies for hydrolyzing plant material mostly rely on the external application of carbohydrate-active enzymes (CAZymes). Alternatively, production organisms can be engineered to secrete CAZymes to reduce the reliance on externally added enzymes. Plant-pathogenic fungi have a vast repertoire of hydrolytic enzymes to sustain their lifestyle, but expression of the corresponding genes is usually highly regulated and restricted to the pathogenic phase. Here, we present a new strategy in using the biotrophic smut fungus Ustilago maydis for the degradation of plant cell wall components by activating its intrinsic enzyme potential during axenic growth. This fungal model organism is fully equipped with hydrolytic enzymes, and moreover, it naturally produces value-added substances, such as organic acids and biosurfactants. To achieve the deregulated expression of hydrolytic enzymes during the industrially relevant yeast-like growth in axenic culture, the native promoters of the respective genes were replaced by constitutively active synthetic promoters. This led to an enhanced conversion of xylan, cellobiose, and carboxymethyl cellulose to fermentable sugars. Moreover, a combination of strains with activated endoglucanase and β-glucanase increased the release of glucose from carboxymethyl cellulose and regenerated amorphous cellulose, suggesting that mixed cultivations could be a means for degrading more complex substrates in the future. In , this proof of principle demonstrates the potential applicability of activating the expression of native CAZymes from phytopathogens in a biocatalytic process.IMPORTANCE This study describes basic experiments that aim at the degradation of plant cell wall components by the smut fungus Ustilago maydis . As a plant pathogen, this fungus contains a set of lignocellulose-degrading enzymes that may be suited for biomass degradation. However, its hydrolytic enzymes are specifically expressed only during plant infection. Here, we provide the proof of principle that these intrinsic enzymes can be synthetically activated during the industrially relevant yeast-like growth. The fungus is known to naturally synthesize valuable compounds, such as itaconate or glycolipids. Therefore, it could be suited for use in a consolidated bioprocess in which more complex and natural substrates are simultaneously converted to fermentable sugars and to value-added compounds in the future.
机译：植物生物质对综合生物过程中宝贵产品的微生物转化可以大大提高生物遗料的生态和经济影响。水解植物材料的目前的策略主要依赖于碳水化合物 - 活性酶（雄虫）的外部施用。或者，可以设计生产生物以分泌巨蜥以减少对外部添加酶的依赖。植物致病性真菌具有巨大的水解酶，以维持其生活方式，但相应基因的表达通常是高度调节和限于致病期的。在这里，我们通过在轴轴生长期间激活其固有酶潜力来展示使用生物养学粉碎真菌Ustilago MASDIS的新策略。这种真菌模型的生物体完全配备有水解酶，而且，它自然产生增值的物质，例如有机酸和生物表面活性剂。为了在轴烯培养的工业相关的酵母状生长期间实现水解酶的放溶性表达，通过组成型活性合成启动子取代各基因的天然启动子。这导致了Xylan，Cellobiose和羧甲基纤维素的转化为可发酵的糖。此外，具有活化的内切葡聚糖酶和β-葡聚糖酶的菌株的组合增加了来自羧甲基纤维素和再生无定形纤维素的葡萄糖的释放，表明混合培养可能是在未来降解更复杂的基材的方法。在这种原则证明中证明了激活生物催化过程中原生婴儿病原体的潜在适用性。分析本研究描述了旨在通过Smut Fungus ustilago maydis降解植物细胞壁组分的基本实验。作为一种植物病原体，该真菌含有一组可适合生物质降解的一组木质纤维素降解酶。然而，其水解酶仅在植物感染期间特别表达。在这里，我们提供原理的证据，即这些本征酶可以在工业相关的酵母状生长期间合成活化。已知真菌自然地合成有价值的化合物，例如衣康酸酯或糖脂。因此，它可以适用于综合生物处理，其中更复杂和天然的基材同时转化为可发酵的糖和未来增值的化合物。

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