Zhang Xiaoling, Miao Qin, Tang Bingling, Mijakovic Ivan, Ji Xiao-Jun, Qu Lingbo, Wei Yongjun
School of Pharmaceutical Sciences, Laboratory of Synthetic Biology, Zhengzhou University, Zhengzhou, 450001, China.
Division of Systems and Synthetic Biology, Department of Biology and Biological Engineering, Chalmers University of Technology, Gothenburg, 41296, Sweden.
Biotechnol Biofuels Bioprod. 2023 Nov 27;16(1):182. doi: 10.1186/s13068-023-02435-8.
Xylo-oligosaccharides (XOS) are considered as a promising type of prebiotics that can be used in foods, feeds, and healthcare products. Xylanases play a key role in the production of XOS from xylan. In this study, we conducted a metagenomic analysis of the fecal microbiota from dairy cows fed with different types of fodders. Despite the diversity in their diets, the main phyla observed in all fecal microbiota were Firmicutes and Bacteroidetes. At the genus level, one group of dairy cows that were fed probiotic fermented herbal mixture-containing fodders displayed decreased abundance of Methanobrevibacter and increased growth of beneficial Akkermansia bacteria. Additionally, this group exhibited a high microbial richness and diversity. Through our analysis, we obtained a comprehensive dataset comprising over 280,000 carbohydrate-active enzyme genes. Among these, we identified a total of 163 potential xylanase genes and subsequently expressed 34 of them in Escherichia coli. Out of the 34 expressed genes, two alkaline xylanases with excellent temperature stability and pH tolerance were obtained. Notably, CDW-xyl-8 exhibited xylanase activity of 96.1 ± 7.5 U/mg protein, with an optimal working temperature of 55 ℃ and optimal pH of 8.0. CDW-xyl-16 displayed an activity of 427.3 ± 9.1 U/mg protein with an optimal pH of 8.5 and an optimal temperature at 40 ℃. Bioinformatic analyses and structural modeling suggest that CDW-xyl-8 belongs to GH10 family xylanase, and CDW-xyl-16 is a GH11 family xylanase. Both enzymes have the ability to hydrolyze beechwood xylan and produce XOS. In conclusion, this metagenomic study provides valuable insights into the fecal microbiota composition of dairy cows fed different fodder types, revealing main microbial groups and demonstrating the abundance of xylanases. Furthermore, the characterization of two novel xylanases highlights their potential application in XOS production.
木寡糖(XOS)被认为是一种很有前景的益生元类型,可用于食品、饲料和保健品中。木聚糖酶在从木聚糖生产XOS的过程中起着关键作用。在本研究中,我们对饲喂不同类型饲料的奶牛粪便微生物群进行了宏基因组分析。尽管它们的饮食存在差异,但在所有粪便微生物群中观察到的主要门类是厚壁菌门和拟杆菌门。在属水平上,一组饲喂含益生菌发酵草药混合物饲料的奶牛,其甲烷短杆菌的丰度降低,有益的阿克曼氏菌的生长增加。此外,该组表现出较高的微生物丰富度和多样性。通过我们的分析,我们获得了一个包含超过280,000个碳水化合物活性酶基因的综合数据集。其中,我们总共鉴定出163个潜在的木聚糖酶基因,随后在大肠杆菌中表达了其中的34个。在这34个表达的基因中,获得了两种具有优异温度稳定性和pH耐受性的碱性木聚糖酶。值得注意的是,CDW-xyl-8的木聚糖酶活性为96.1±7.5 U/mg蛋白质,最佳工作温度为55℃,最佳pH为8.0。CDW-xyl-16的活性为427.3±9.1 U/mg蛋白质,最佳pH为8.5,最佳温度为40℃。生物信息学分析和结构建模表明,CDW-xyl-8属于GH10家族木聚糖酶,CDW-xyl-16是GH11家族木聚糖酶。这两种酶都有能力水解山毛榉木聚糖并产生XOS。总之,这项宏基因组研究为饲喂不同饲料类型的奶牛粪便微生物群组成提供了有价值的见解,揭示了主要微生物群体并证明了木聚糖酶的丰度。此外,两种新型木聚糖酶的特性突出了它们在XOS生产中的潜在应用。