Zhu Ning, Yang Jinshui, Ji Lei, Liu Jiawen, Yang Yi, Yuan Hongli
State Key Laboratory of Agrobiotechnology, College of Biological Sciences, Beijing, China.
State Key Laboratory of Agrobiotechnology, College of Biological Sciences, Beijing, China ; National Energy R & D Center for Non-food Biomass, China Agricultural University, Beijing, 100193 China.
Biotechnol Biofuels. 2016 Nov 9;9:243. doi: 10.1186/s13068-016-0658-z. eCollection 2016.
Microbial consortia represent promising candidates for aiding in the development of plant biomass conversion strategies for biofuel production. However, the interaction between different community members and the dynamics of enzyme complements during the lignocellulose deconstruction process remain poorly understood. We present here a comprehensive study on the community structure and enzyme systems of a lignocellulolytic microbial consortium EMSD5 during growth on corn stover, using metagenome sequencing in combination with quantitative metaproteomics.
The taxonomic affiliation of the metagenomic data showed that EMSD5 was primarily composed of members from the phyla , and . The carbohydrate-active enzyme (CAZyme) annotation revealed that representatives of encoded a broad array of enzymes responsible for hemicellulose and cellulose deconstruction. Extracellular metaproteome analysis further pinpointed the specific role and synergistic interaction of populations in plant polysaccharide breakdown. In particular, a wide range of xylan degradation-related enzymes, including xylanases, β-xylosidases, α-l-arabinofuranosidases, α-glucuronidases and acetyl xylan esterases, were secreted by diverse members from during growth on corn stover. Using label-free quantitative proteomics, we identified the differential secretion pattern of a core subset of enzymes, including xylanases and cellulases with multiple carbohydrate-binding modules (CBMs). In addition, analysis of the coordinate expression patterns indicated that transport proteins and hypothetical proteins may play a role in bacteria processing lignocellulose. Moreover, enzyme preparation from EMSD5 demonstrated synergistic activities in the hydrolysis of pretreated corn stover by commercial cellulases from .
These results demonstrate that the corn stover-adapted microbial consortium EMSD5 harbors a variety of lignocellulolytic anaerobic bacteria and degradative enzymes, especially those implicated in hemicellulose decomposition. The data in this study highlight the pivotal role and cooperative relationship of members in the biodegradation of plant lignocellulose by EMSD5. The differential expression patterns of enzymes reveal the strategy of sequential lignocellulose deconstruction by EMSD5. Our findings provide insights into the mechanism by which consortium members orchestrate their array of enzymes to degrade complex lignocellulosic biomass.
微生物群落有望助力开发用于生物燃料生产的植物生物质转化策略。然而,在木质纤维素解构过程中,不同群落成员之间的相互作用以及酶补充物的动态变化仍知之甚少。我们在此展示一项关于木质纤维素分解微生物群落EMSD5在玉米秸秆上生长期间的群落结构和酶系统的全面研究,该研究采用宏基因组测序与定量宏蛋白质组学相结合的方法。
宏基因组数据的分类归属显示,EMSD5主要由来自门、和的成员组成。碳水化合物活性酶(CAZyme)注释表明,编码了一系列负责半纤维素和纤维素解构的酶。细胞外宏蛋白质组分析进一步明确了群体在植物多糖分解中的具体作用和协同相互作用。特别是,在玉米秸秆上生长期间,不同成员分泌了多种与木聚糖降解相关的酶,包括木聚糖酶、β-木糖苷酶、α-L-阿拉伯呋喃糖苷酶、α-葡萄糖醛酸酶和乙酰木聚糖酯酶。使用无标记定量蛋白质组学,我们确定了包括具有多个碳水化合物结合模块(CBM)的木聚糖酶和纤维素酶在内的核心酶子集的差异分泌模式。此外,对协同表达模式的分析表明,转运蛋白和假定蛋白可能在细菌处理木质纤维素中发挥作用。此外,EMSD5的酶制剂在与来自的商业纤维素酶共同水解预处理玉米秸秆时表现出协同活性。
这些结果表明,适应玉米秸秆的微生物群落EMSD5含有多种木质纤维素分解厌氧细菌和降解酶,尤其是那些与半纤维素分解有关的酶。本研究中的数据突出了成员在EMSD5对植物木质纤维素生物降解中的关键作用和合作关系。酶的差异表达模式揭示了EMSD5对木质纤维素进行顺序解构的策略。我们的研究结果为群落成员协调其酶阵列以降解复杂木质纤维素生物质的机制提供了见解。