Development of a pipeline for High Throughput Screening of Functional Metagenomic Library in Search for Novel Lipolytic Enzymes

PhD Dissertation Defense
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Location
Building 3, Level 5, Room 5209

Abstract

The demand for a 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, the 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 constructed metagenomics library of the Red Seawater. 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.

Brief Biography

Amani Almaabadi is a Ph.D. candidate in Takashi Gojobori's group at King Abdullah University of Science and Technology (KAUST) Thuwal, Saudi Arabia. She holds her Master's Degree in Bioscience, (KAUST), Thuwal, Saudi Arabia, and a Bachelors degree in Biochemistry, (KAU), College of Science, Jeddah, Saudi Arabia. Her research interests broadly concern ​Marine metagenomics, Environmental microbiology, biotechnology, synthetic biology, and industrial enzymes.