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具有高木聚糖酶活性的新型木质纤维素降解复合微生物系统的功能特征和多样性。

Functional characteristics and diversity of a novel lignocelluloses degrading composite microbial system with high xylanase activity.

机构信息

College of Agronomy and Biotechnology/Center of Biomass Engineering, China Agricultural University, No. 2 Yuanmingyuan West Road, Haidian District, Beijing 100193, China.

出版信息

J Microbiol Biotechnol. 2010 Feb;20(2):254-64.

Abstract

To obtain an efficient natural lignocellulolytic complex enzyme, we screened an efficient lignocellulose degrading composite microbial system (XDC-2) from composted agricultural and animal wastes amended soil following a long-term directed acclimation. The XDC-2 could not only degrade natural lignocelluloses, but could also secret extracellular xylanase efficiently in liquid culture under static conditions at room temperature. The XDC-2 degraded rice straw by 60.3% after fermentation for 15 days. Hemicelluloses were decomposed effectively, while the extracellular xylanase activity was dominant with an activity of 8.357 U ml-1 on day 6 of the fermentation period. The extracellular crude enzyme noticeably hydrolyzed natural lignocelluloses. The optimum temperature and pH for the xylanase activity were 40 degrees and 6.0. However, the xylanase was activated in a wide pH range of 3.0-10.0, and retained more than 80% of its activity at 25-35 degrees and pH 5.0-8.0 after three days of incubation in liquid culture under static conditions. PCR-DGGE analysis of successive subculture indicated that the XDC-2 was structurally stable over long-term restricted and directed cultivation. Analysis of 16S rRNA gene clone library showed that the XDC-2 was mainly composed of mesophilic bacteria related to the genera Clostridium, Bacteroides, Alcaligenes, Pseudomonas, etc. Our results offer a new approach to exploring efficient lignocellulolytic enzymes by constructing a high-performance composite microbial system with synergistic complex enzymes.

摘要

为了获得高效的天然木质纤维素复合酶,我们从长期定向驯化的堆肥农业和动物废物改良土壤中筛选出一种高效的木质纤维素降解复合微生物系统(XDC-2)。XDC-2 不仅可以降解天然木质纤维素,而且可以在室温下的静态液体培养中高效分泌胞外木聚糖酶。XDC-2 在发酵 15 天后可将稻草降解 60.3%。半纤维素被有效分解,而胞外木聚糖酶活性在发酵期第 6 天占主导地位,活性为 8.357 U ml-1。胞外粗酶明显水解天然木质纤维素。木聚糖酶活性的最适温度和 pH 值分别为 40 度和 6.0。然而,木聚糖酶在 pH 值 3.0-10.0 的宽范围内被激活,并且在 25-35 度和 pH 值 5.0-8.0 的条件下在静态液体培养中孵育三天后保留超过 80%的活性。连续继代培养的 PCR-DGGE 分析表明,XDC-2 在长期限制和定向培养下结构稳定。16S rRNA 基因克隆文库分析表明,XDC-2 主要由与梭菌属、拟杆菌属、产碱杆菌属、假单胞菌属等相关的嗜温菌组成。我们的结果为通过构建具有协同复合酶的高性能复合微生物系统来探索高效木质纤维素酶提供了一种新方法。

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