Exploration of Lipase Enzyme from Soil through Metagenomic Approach

Abstract

This research aimed to explore lipase enzyme from soil through metagenomic approach. The DNA was extracted directly from the topsoil and river sediment by SDS-based lysis method. The soil genomic DNA was sheared into approximately 40-kb fragments The construction of genomic library was carried out by cloning the fragment DNA into  fosmid pCC2FOS vector and host E. coli EPI300-T1R accordance with the protocols of CopyControl™ Fosmid production library kit. Screening of lipase-producing clones was performed based on its lipolytic activity using Luria-Bertani agar plate 12.5 µg/ ml chloramphenicol, 10 µg/ml Rhodamine B and 1 % olive oil as a substrate. The assay plates were incubated at 550 C. The lipolytic activity was identified as an orange around colonies. There were 5 (five) clones of 32 screened-clones that expressed lipolytic activity. The next research will characterize the enzyme and determine the sequence of the enzyme-encoding gene.

Keywords: Lipase, soil, metagenomic, fosmid vector, pCC2FOS, E. coli EPI300-T1R