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Enhancement of β-xylosidase productivity in cellulase producing fungus Acremonium cellulolyticus

机译:提高产纤维素酶的真菌纤维素解孢菌中β-木糖苷酶生产力

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Enzymatic hydrolysis is one of the most important processes in bioethanol production from lignocellulosic biomass. Acremonium cellulolyticus is a filamentous fungus with high cellulase production but productivity of hemicellulase, especially β-xylosidase, is lower than other filamentous fungi. We identified 2.4 Kb β-xylosidase gene in the A. cellulolyticus genome sequence information and it encoded 798 amino acids without introns. To enhance hemicellulase productivity in A. cellulolyticus, we transformed this fungus with the identified β-xylosidase gene driven by the cellobiohydrolase Ι (cbh1) promoter, using the protoplast-polyethyleneglycol (PEG) method, and obtained a transformant, YKX1. Hydrolysis rate of xylooligosaccharides was more than 50-fold higher using culture supernatant from YKX1 than that from the parental strain, Y-94. Total cellulase activity (measured by filter paper assay) in YKX1 was not affected by the cbh1 promoter used for expression of β-xylosidase, and induced by cellulose. Since YKX1 can produce larger amount of β-xylosidase without affecting cellulase productivity, it is considered to be beneficial for practical monosaccharide recoveries from lignocellulosic biomass.
机译:酶促水解是从木质纤维素生物质生产生物乙醇中最重要的过程之一。顶头纤维素酶是一种高产纤维素酶的丝状真菌,但半纤维素酶,特别是β-木糖苷酶的生产率低于其他丝状真菌。我们在A. cellulolyticus基因组序列信息中鉴定了2.4 Kbβ-木糖苷酶基因,它编码了798个没有内含子的氨基酸。为了提高溶纤曲霉中的半纤维素酶生产率,我们使用原生质体-聚乙二醇(PEG)方法用由纤维二糖水解酶I(cbh1)启动子驱动的已鉴定的β-木糖苷酶基因转化了这种真菌,并获得了转化子YKX1。使用来自YKX1的培养上清液比来自亲本菌株Y-94的木糖低聚糖的水解速率高50倍以上。 YKX1中的总纤维素酶活性(通过滤纸测定法测定)不受用于表达β-木糖苷酶的cbh1启动子的影响,并由纤维素诱导。由于YKX1可以产生大量的β-木糖苷酶而不影响纤维素酶的生产率,因此被认为对于从木质纤维素生物质中回收实际单糖是有益的。

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