Purification and properties of a lipase from Cephaloleia presignis (Coleoptera, Chrysomelidae)

Abstract

A novel lipase from the insect Cephaloleia presignis was purified by a procedure involving ammonium sulphate precipitation, and Phenyl Toyopearl 650M, DEAE-5PW and hydrophobic-interaction column chromatographies. The purified lipase was homogeneous with a molecular mass of 31000 Da by SDS/PAGE and of 29000 Da by gel filtration on a Superose 12 column. The enzyme was identified as a glycoprotein with a pI of 6.9. The enzyme unspecifically liberated short-chain to long-chain fatty acids from p-nitrophenyl esters, methyl esters and triglycerides. The N-terminal 28 amino acid residues were determined as AGTLGYATRHVLPIFTLDDYTGSNEMWG, which showed no similarity with known proteins, suggesting that the purified lipase may belong to a novel class of hydrolases.

Purification and Biochemical Characterization of Digestive Lipase in Whiteleg Shrimp

“…PVL does not rely on calcium for activity, indicating that PVL catalytic properties are dissimilar to mammalian lipases. This is supported by reports showing that lipase in insects R. prolixus (Grillo et al 2007) and C. presignis (Arreguín-Espinosa et al 2000) have the same activity regardless of the calcium concentration. To date, it is not certain that invertebrate digestive lipases require a moderate concentration of calcium for activity or stability, as observed in mammalian digestive lipases.…”

Comparative and functional genomics of lipases in holometabolous insects

“…Although TAGhydrolysing lipases have been isolated from lepidopteran, coleopteran and dipteran species, these either belong to other lipase families or have no known sequence (Arrese and Wells, 1994;Arreguin-Espinosa et al, 2000;Grö nke et al, 2005;Arrese et al, 2006). Moreover other differences in lipolytic machinery might mean that the presentation of TAG substrates to insect lipases do not require precisely the same lid and loop motifs as are needed in mammalian lipases (see Section 3.5.1 below).…”

Lipazės aliejų perdirbimui. Eksperimentinė kai kurių fermentų ir substratų atranka biodyzelino komponentų gamybai

“…2). Lipases are usually shown to be specific to substrates of both medium [10] and long [15] chains.…”