Medline/PubMed Citation | Related Articles in PubMed | Download to Citation Manager

Biotechnology and Applied Biochemistry (2000) 31, (239–244) (Printed in Great Britain)

PDF

Legacy HTML

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

Roberto Arreguín-Espinosa*¹, Barbarín Arreguín* and Carolina González†

*Instituto de Química, UNAM, Circuito Exterior, Ciudad Universitaria, Coyoacán 04510 México, D.F., and †Facultad de Ciencias Químicas, UANL, Pedro de Alba S/N, 66451 , Nuevo Leon, México

Abbreviations used: IEF, isoelectric focusing; DFP di-isopropyl fluorophosphate; pNP-ester, p-nitrophenyl ester.

¹ To whom correspondence should be addressed.

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 indentified 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.

Received 7 September 1999/2 February 2000; accepted 13 February 2000

Portland Press Ltd © 2000