Zhou Man, Guo Peng, Wang Tao, Gao Lina, Yin Huijun, Cai Cheng, Gu Jie, Lü Xin
College of Food Science and Engineering, Northwest A&F University, Yangling, Shaanxi Province China.
College of Information Engineering, Northwest A&F University, Yangling, Shaanxi Province China.
Biotechnol Biofuels. 2017 Aug 22;10:198. doi: 10.1186/s13068-017-0885-y. eCollection 2017.
Degradation of pectin in lignocellulosic materials is one of the key steps for biofuel production. Biological hydrolysis of pectin, i.e., degradation by pectinolytic microbes and enzymes, is an attractive paradigm because of its obvious advantages, such as environmentally friendly procedures, low in energy demand for lignin removal, and the possibility to be integrated in consolidated process. In this study, a metagenomics sequence-guided strategy coupled with enrichment culture technique was used to facilitate targeted discovery of pectinolytic microbes and enzymes. An apple pomace-adapted compost (APAC) habitat was constructed to boost the enrichment of pectinolytic microorganisms.
Analyses of 16S rDNA high-throughput sequencing revealed that microbial communities changed dramatically during composting with some bacterial populations being greatly enriched. Metagenomics data showed that apple pomace-adapted compost microbial community (APACMC) was dominated by and . Functional analysis and carbohydrate-active enzyme profiles confirmed that APACMC had been successfully enriched for the targeted functions. Among the 1756 putative genes encoding pectinolytic enzymes, 129 were predicted as novel (with an identity <30% to any CAZy database entry) and only 1.92% were more than 75% identical with proteins in NCBI environmental database, demonstrating that they have not been observed in previous metagenome projects. Phylogenetic analysis showed that APACMC harbored a broad range of pectinolytic bacteria and many of them were previously unrecognized.
The immensely diverse pectinolytic microbes and enzymes found in our study will expand the arsenal of proficient degraders and enzymes for lignocellulosic biofuel production. Our study provides a powerful approach for targeted mining microbes and enzymes in numerous industries.
木质纤维素材料中果胶的降解是生物燃料生产的关键步骤之一。果胶的生物水解,即通过果胶分解微生物和酶进行降解,是一种有吸引力的模式,因为它具有明显的优势,如环境友好的程序、去除木质素所需的低能量需求以及有可能整合到联合工艺中。在本研究中,采用宏基因组学序列引导策略结合富集培养技术,以促进果胶分解微生物和酶的靶向发现。构建了一个适应苹果渣的堆肥(APAC)生境,以促进果胶分解微生物的富集。
16S rDNA高通量测序分析表明,在堆肥过程中微生物群落发生了显著变化,一些细菌种群大量富集。宏基因组学数据显示,适应苹果渣的堆肥微生物群落(APACMC)以[具体菌群1]和[具体菌群2]为主。功能分析和碳水化合物活性酶谱证实,APACMC已成功富集到靶向功能。在1756个推定的编码果胶分解酶的基因中,有129个被预测为新基因(与任何CAZy数据库条目同一性<30%),只有1.92%与NCBI环境数据库中的蛋白质同一性超过75%,这表明它们在以前的宏基因组项目中未被观察到。系统发育分析表明,APACMC含有广泛的果胶分解细菌,其中许多以前未被识别。
我们的研究中发现的极其多样的果胶分解微生物和酶将扩大用于木质纤维素生物燃料生产的高效降解菌和酶的库。我们的研究为众多行业靶向挖掘微生物和酶提供了一种强大的方法。