Metagenomics-Based Cloning of Amilase-Encoding Genes from the Uncultured Symbiotic Bacteria of a Marine Sponge Theonella swinhoei from Kapoposang Island, South Sulawesi
Abstract: Marine sponges have
recently been recognized as the source ofenzymes, including members of
hydrolases. Hydrolytic enzymes are extracellularly produced by
sponge-associated bacteria to mediate the metabolism of complex organic
matters, thereby assisting the sponge hosts in nutrition and metabolic
processes. Among hydrolytic enzymes, amilaseshas attracted increasing attention
due to their potential industrial applications. This research work was aimed
atutilizing functional metagenomicsapproach for the discovery of amilases
derived from the uncultured symbiotic bacteriaof the Indonesian marine sponge
Theonella swinhoei. Weinitially constructed a small-insert metagenomiclibrary
in Escherichia coliby cloning of metagenome in the size range of5-20 kb
prepared from the sponge’s microbiome. Further functional screening of the
resulting metagenomic library led to the isolation of two recombinant E. coli
clones potentially harboring amilase genes, as indicated by the presence of
clearing zones surroinding the selective medium containing 1% amilum.
Keywords: Theonella swinhoei;
Uncultured symbiotic bacteria; DNA cloning; amylase gene
Author: Franciscus Edi
Priyono, Dewi Seswita Zilda, Yudi Kusnadi, Tri A Hadi, Irvina Nurrachmi,
Agustinus Robert Uria
Journal Code: jpperikanangg170001
