CSIC, Institute of Catalysis, Department of Applied Biocatalysis, Madrid, Spain.
Microb Biotechnol. 2009 Jan;2(1):31-9. doi: 10.1111/j.1751-7915.2008.00057.x. Epub 2008 Oct 13.
In recent years, the application of approaches for harvesting DNA from the environment, the so-called, 'metagenomic approaches' has proven to be highly successful for the identification, isolation and generation of novel enzymes. Functional screening for the desired catalytic activity is one of the key steps in mining metagenomic libraries, as it does not rely on sequence homology. In this mini-review, we survey high-throughput screening tools, originally developed for directed evolution experiments, which can be readily adapted for the screening of large libraries. In particular, we focus on the use of in vitro compartmentalization (IVC) approaches to address potential advantages and problems the merger of culture-independent and IVC techniques might bring on the mining of enzyme activities in microbial communities.
近年来,从环境中提取 DNA 的方法(即所谓的“宏基因组学方法”)在鉴定、分离和产生新型酶方面已被证明非常成功。在挖掘宏基因组文库时,针对所需催化活性的功能筛选是关键步骤之一,因为它不依赖于序列同源性。在这篇小型综述中,我们调查了高通量筛选工具,这些工具最初是为定向进化实验开发的,可轻松适用于大型文库的筛选。特别是,我们重点讨论了体外分隔(IVC)方法的使用,以解决在挖掘微生物群落中的酶活性时,将非培养技术与 IVC 技术结合可能带来的潜在优势和问题。