Cellulase-producing microorganisms are considered as a key player in various environments to degrade the plant biomass and were isolated from various environment like soils, mangroves and oceans. The Red Sea has a unique environment in terms of high seawater temperature, high salinity and low nutrients. This study aims of examining if the Red Sea is a potential resource for cellulase-producing microorganisms and cellulase genes. First, I investigated types of microbial cellulase genes in the Red Sea based on public metagenomic datasets. The analysis revealed 3,383 microbial cellulase were more abundant in shallow depth than in deep seawater, and were classified into 16 sub-GH orthologous groups. These results suggest that the Red Sea environment is potentially an excellent gene resource of microbial cellulases due to its high diversity. Next, cellulase-producing microorganisms were isolated and screened from the Red Sea. Three bacterial and one fungal strain were successfully obtained. The MLTS analysis showed that the three bacterial strains belong to Bacillus paralichiniformis. The 18S rRNA of fungal strain showed 99% similarity to Aspergillus ustus and the enzymatic assay of the four strains showed high cellulase activity. These results suggest that these four isolates secreted active cellulases. Next, I tried to identify cellulase genes actually working during their cellulolysis by conducting comparative transcriptome analysis of the candidate genes and identified cellulase genes that are highly expressed during cellulolysis. To my knowledge, it is the first attempt to find out cellulase genes functioning during their cellulolysis among distinct cellulases on genomes of microorganisms. The results showed that although all the candidate genes were upregulated in general, a limited number of cellulase genes were highly expressed, which are highly expected to have a crucial role in cellulolysis. I also identified operon structures composed of genes including cellulases. This will provide us with the information to elucidate the cellular mechanisms occurring along with the cellulolysis in bacterial strains. We can expect that the Red Sea is a potential resource for new cellulase genes applicable for the industry. These information can be significantly useful for the bio-prospecting research of microbial cellulases in the Red Sea.
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|KAUST Research Repository