New cold-adapted lipase from Photobacterium lipolyticum sp. nov. that is closely related to filamentous fungal lipases

Abstract

A Photobacterium strain, M37, showing lipolytic activity, was previously isolated from an intertidal flat of the Yellow Sea in Korea and identified as Photobacterium lipolyticum sp. nov. In the present study, the corresponding gene was cloned using the shotgun method. The amino acid sequence deduced from the nucleotide sequence (1,023 bp) corresponded to a protein of 340 amino acid residues with a molecular weight of 38,026. No sequence similarity was found with any known bacterial lipases/esterases; instead, the most similar enzymes were several filamentous fungal lipases. Although the similarity was very low (less than 16%), there were many conserved regions over the entire sequence and N-terminal oxyanion hole (RG) region, a signature sequence of filamentous fungal lipases. The novel protein M37 was produced in both a soluble and insoluble form when the Escherichia coli cells harboring the gene were cultured at 18 degrees C. The soluble protein exhibited lipase activity in a pH-stat assay using an olive oil emulsion. The M37 lipase also displayed a maximum activity at 25 degrees C and maintained its activity at a low temperature range (5-25 degrees C) with an activation energy (E(a)) of 2.07 kcal/mol. Accordingly, these results indicate that the M37 lipase from P. lipolyticum sp. nov. is a new cold-adapted enzyme.

Isolation and Characterization of a Novel Lipase from a Metagenomic Library of Tidal Flat Sediments: Evidence for a New Family of Bacterial Lipases

“…All of the filamentous fungal lipases, including R. miehei lipase, which was the most similar to LipG, have Arg-Gly oxyanion residues in the N-terminal part, whereas most bacterial and other fungal lipases have His-Gly residues (22). In fact, this Arg-Gly oxyanion hole sequence is known to be a unique signature sequence for filamentous fungal lipases.…”

“…In fact, this Arg-Gly oxyanion hole sequence is known to be a unique signature sequence for filamentous fungal lipases. Unlike other bacterial lipases, the oxyanion hole sequence of a lipase recently cloned from Photobacterium lipolyticum also contained the Arg-Gly sequence (22). Interestingly, P. lipolyticum has been isolated from tidal flat sediments.…”

A novel cold-adapted lipase, LP28, from a mesophilic Streptomyces strain

“…Strain KB700A [22], Photobacterium lipolytic sp. nov [23], and Pseudoalteromonas haloplanktis TAC125 [24]. LP28 hydrolyzed triolein and released 1,3-diolein, 1, 2-diolein and oleic acid as the end products (Fig.…”

Synthesis of trimethylolpropane esters of oleic acid by Lipoprime 50T

“…The hydrolytic effect of lipase on p-NPB solution in 2-propanol [5,9] was investigated by measuring the change of optical density at 400-410 nm within 3-6 min at 30°C and pH 7.0-10.0, 100 mM universal buffer (Britton-Robinson buffer, composed of acetic, ortho-boric and orthophosphoric acids at a ratio of 1:1:1 providing buffering capacity over a wide range of pH) [2,10].…”