Development of a Pipeline for Single Cell Microfluidics Screening of Metagenomic Library for Finding Novel Lipolytic Enzymes

Abstract

The demand for novel and robust microbial biocatalysts for industrial and pharmaceutical applications continue to grow at a fast pace.This warrants a continuous need for advanced tools and technologies to exploit the vast metabolic potential of microorganisms in different environments. Unlike culture-based studies that can only reveal the metabolic potential of cultivable microorganisms, functional metagenomics charts the enzymatic potential of the entire microbial communities in a given environment. This method has substantially contributed to the effective discovery of unique microbial genes for industrial and medical applications. Functional metagenomics involves the extraction of microbial DNA directly from environmental samples,construction of an expression library containing the entire microbial genome, and screening the libraries for the presence of desired phenotypes. Therefore, development of a pipeline for analyzing and screening metagenomic libraries is essential for rapid detection of the desired features from thousands of clones of a single library. Here, we developed a pipeline for high-throughput screening of the lipolytic genes from the Red Sea.Further, a high-throughput single cell microfluidics platform combined with a laser-based fluorescent screening bioassay was deployed to discover new lipolytic genes. Our analysis led to the identification of 24 microbial genes for lipases and esterase from a metagenomic library of the Red Sea water. The results further showed that the constructed pipeline is robust in conducting functional metagenomics and for the discovery of new genes. It also implies that the Red Sea is a rich under- investigated source of natural resources of new genes and gene products.