Kunath Benoit J, Bremges Andreas, Weimann Aaron, McHardy Alice C, Pope Phillip B
Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, 5003, 1432, Ås, Norway.
Computational Biology of Infection Research, Helmholtz Centre for Infection Research, 38124, Braunschweig, Germany.
Methods Mol Biol. 2017;1588:255-277. doi: 10.1007/978-1-4939-6899-2_20.
Microorganisms play a primary role in regulating biogeochemical cycles and are a valuable source of enzymes that have biotechnological applications, such as carbohydrate-active enzymes (CAZymes). However, the inability to culture the majority of microorganisms that exist in natural ecosystems using common culture-dependent techniques restricts access to potentially novel cellulolytic bacteria and beneficial enzymes. The development of molecular-based culture-independent methods such as metagenomics enables researchers to study microbial communities directly from environmental samples, and presents a platform from which enzymes of interest can be sourced. We outline key methodological stages that are required as well as describe specific protocols that are currently used for metagenomic projects dedicated to CAZyme discovery.
微生物在调节生物地球化学循环中发挥着主要作用,并且是具有生物技术应用的酶(如碳水化合物活性酶,CAZymes)的宝贵来源。然而,使用常见的依赖培养的技术无法培养自然生态系统中存在的大多数微生物,这限制了获取潜在新型纤维素分解细菌和有益酶的途径。基于分子的非培养方法(如宏基因组学)的发展使研究人员能够直接从环境样品中研究微生物群落,并提供了一个可以获取感兴趣酶的平台。我们概述了所需的关键方法步骤,并描述了目前用于致力于发现CAZyme的宏基因组项目的具体方案。