Cloning and Characterization of Lipase Gene from a Local Isolate of Pseudoxanthomonas sp.

Abstract

Lipase gene from Pseudoxanthomonas sp. was cloned through in vitro amplification from total chromosomal DNA. The gene was sequenced and characterized, coding for 312 amino acid residues. Homological analysis showed that the gene has 98% similarity to lipolytic gene from Uncultured Pseudomonas sp (GenBank No. AKA58891.1). Further analysis appeared that the sequences showed similar unique motifs of lipase sub-family I.1, such as pentapeptide (GHSHG) motif, tetrapeptide (GMLG) motif, and catalytic triad. In additional, 3D structure analysis based on crystal structure of Pseudomonas aeruginose (PDB ID 1ex9) showed that both structure of lipases are similar except on the conformation of catalytic residue of His277 showing to shift more far away compared to that the control.

Cloning, heterologous expression, and characterization of a novel thioesterase from natural sample

“…Some thermostable lipases and esterases from thermophilic organisms were cloned and characterized, from various natural sources such as hot springs, craters or compost, both through cultivation [14,16,17,18,19,20,21,22] and metagenomics approaches [23]. In this report, cloning, heterologous expression, and characterization of a novel gene from natural sample of Domas Hot spring, West Java, namely TESITB was described.…”

Effect of mutation at oxyanion hole residu (H110F) on activity of Lk4 lipase

“…Exploration of thermostable lipases were extensively carried out [ 19 ]. Various lipases were isolated from local strains and traditional compost [ [20] , [21] , [22] , [23] , [24] ]. One of the lipase gene namely LK_ITB5a ( LK4 ) was previously isolated directly from compost using metagenomic approach at thermogenic phase.…”