Dai Xin, Tian Yan, Li Jinting, Luo Yingfeng, Liu Di, Zheng Huajun, Wang Jiaqi, Dong Zhiyang, Hu Songnian, Huang Li
Appl Environ Microbiol. 2015 Feb;81(4):1375-86. doi: 10.1128/AEM.03682-14.
The bovine rumen represents a highly specialized bioreactor where plant cell wall polysaccharides (PCWPs) are efficiently deconstructed via numerous enzymes produced by resident microorganisms. Although a large number of fibrolytic genes from rumen microorganisms have been identified, it remains unclear how they are expressed in a coordinated manner to efficiently degrade PCWPs. In this study, we performed a metatranscriptomic analysis of the rumen microbiomes of adult Holstein cows fed a fiber diet and obtained a total of 1,107,083 high-quality non-rRNA reads with an average length of 483 nucleotides. Transcripts encoding glycoside hydrolases (GHs) and carbohydrate binding modules (CBMs) accounted for 1% and 0.1% of the total non-rRNAs, respectively. The majority (98%) of the putative cellulases belonged to four GH families (i.e., GH5, GH9, GH45, and GH48) and were primarily synthesized by Ruminococcus and Fibrobacter. Notably, transcripts for GH48 cellobiohydrolases were relatively abundant compared to the abundance of transcripts for other cellulases. Two-thirds of the putative hemicellulases were of the GH10, GH11, and GH26 types and were produced by members of the genera Ruminococcus, Prevotella, and Fibrobacter. Most (82%) predicted oligosaccharide-degrading enzymes were GH1, GH2, GH3, and GH43 proteins and were from a diverse group of microorganisms. Transcripts for CBM10 and dockerin, key components of the cellulosome, were also relatively abundant. Our results provide metatranscriptomic evidence in support of the notion that members of the genera Ruminococcus, Fibrobacter, and Prevotella are predominant PCWP degraders and point to the significant contribution of GH48 cellobiohydrolases and cellulosome-like structures to efficient PCWP degradation in the cow rumen.
牛瘤胃是一个高度专业化的生物反应器,其中植物细胞壁多糖(PCWPs)通过常驻微生物产生的多种酶被有效分解。尽管已鉴定出大量来自瘤胃微生物的纤维分解基因,但它们如何以协调的方式表达以有效降解PCWPs仍不清楚。在本研究中,我们对喂食纤维日粮的成年荷斯坦奶牛的瘤胃微生物群进行了宏转录组分析,共获得1,107,083条高质量非rRNA reads,平均长度为483个核苷酸。编码糖苷水解酶(GHs)和碳水化合物结合模块(CBMs)的转录本分别占非rRNA总数的1%和0.1%。大多数(98%)假定的纤维素酶属于四个GH家族(即GH5、GH9、GH45和GH48),主要由瘤胃球菌属和纤维杆菌属合成。值得注意的是,与其他纤维素酶的转录本丰度相比,GH48纤维二糖水解酶的转录本相对丰富。三分之二的假定半纤维素酶属于GH10、GH11和GH26类型,由瘤胃球菌属、普雷沃氏菌属和纤维杆菌属的成员产生。大多数(82%)预测的寡糖降解酶是GH1、GH2、GH3和GH43蛋白,来自多种微生物。纤维素体的关键成分CBM10和锚定蛋白的转录本也相对丰富。我们的结果提供了宏转录组学证据,支持瘤胃球菌属、纤维杆菌属和普雷沃氏菌属成员是主要的PCWP降解菌这一观点,并指出GH48纤维二糖水解酶和类纤维素体结构对奶牛瘤胃中PCWP的有效降解有重要贡献。