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从红栓菌 PF-2 中提取的纤维素酶的特性及其在生物质糖化中的应用。

Characterization of cellulolytic extract from Pycnoporus sanguineus PF-2 and its application in biomass saccharification.

机构信息

Department of Biochemistry and Molecular Biology, Federal University of Viçosa, Viçosa, Minas Gerais, 36570-000, Brazil.

出版信息

Appl Biochem Biotechnol. 2012 Mar;166(6):1586-603. doi: 10.1007/s12010-012-9565-3. Epub 2012 Feb 11.

Abstract

The aim of this work was to evaluate the biochemical features of the white-rot fungi Pycnoporus sanguineus cellulolytic complex and its utilization to sugarcane bagasse hydrolysis. When cultivated under submerged fermentation using corn cobs as carbon source, P. sanguineus produced high FPase, endoglucanase, β-glucosidase, xylanase, mannanase, α-galactosidase, α-arabinofuranosidase, and polygalacturonase activities. Cellulase activities were characterized in relation to pH and temperature. β-Glucosidase and FPase activities were higher at 55 °C, pH 4.5, and endoglucanase activity was higher at 60 °C, in a pH range of 3.5-4.0. All cellulase activities were highly stable at 40 and 50 °C through 48 h of pre-incubation. Crude enzymatic extract from P. sanguineus was applied in a saccharification experiment using acid-treated and alkali-treated sugarcane bagasse as substrate, and the hydrolysis yields were compared to that obtained by a commercial cellulase preparation. Reducing sugar yields of 60.4% and 64.0% were reached when alkali-treated bagasse was hydrolyzed by P. sanguineus extract and commercial cellulase, respectively. Considering the glucose production, it was observed that P. sanguineus extract and commercial cellulase ensured yields of 22.6% and 36.5%, respectively. The saccharification of acid-treated bagasse was lower than that of alkali-treated bagasse regardless of the cellulolytic extract. The present work showed that P. sanguineus has a great potential as an enzyme producer for biomass saccharification.

摘要

本工作旨在评估白腐真菌血红密孔菌纤维素复合体系的生化特性及其在甘蔗渣水解中的应用。当以玉米芯作为碳源进行深层发酵时,血红密孔菌产生了高的 FPase、内切葡聚糖酶、β-葡萄糖苷酶、木聚糖酶、甘露聚糖酶、α-半乳糖苷酶、α-阿拉伯呋喃糖苷酶和聚半乳糖醛酸酶活性。研究了纤维素酶活性与 pH 和温度的关系。β-葡萄糖苷酶和 FPase 活性在 55°C、pH4.5 下较高,内切葡聚糖酶活性在 60°C、pH3.5-4.0 范围内较高。粗酶提取物在 40 和 50°C 下经过 48 小时的预孵育,所有纤维素酶活性都具有高度稳定性。将血红密孔菌粗酶提取物应用于酸处理和碱处理甘蔗渣作为底物的糖化实验,并将水解产率与商业纤维素酶制剂的水解产率进行比较。用血红密孔菌粗酶提取物和商业纤维素酶分别水解碱处理蔗渣,还原糖产率分别达到 60.4%和 64.0%。考虑到葡萄糖的产量,观察到血红密孔菌粗酶提取物和商业纤维素酶分别保证了 22.6%和 36.5%的产率。无论使用哪种纤维素酶提取物,酸处理蔗渣的糖化都低于碱处理蔗渣。本研究表明,血红密孔菌具有作为生物量糖化用酶制剂生产的巨大潜力。

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