caa a22 4500 44531902X CHVBK 20180317142657.0 cr unu---uuuuu 170323e20110401xx s 000 0 eng 10.1007/s10126-010-9298-7 doi (NATIONALLICENCE)springer-10.1007/s10126-010-9298-7 Purification and Biochemical Characterization of Digestive Lipase in Whiteleg Shrimp [Elektronische Daten] [Crisalejandra Rivera-Pérez, Fernando García-Carreño, Reinhard Saborowski] Penaeus vannamei lipase was purified from midgut gland of whiteleg shrimp. Pure lipase (E.C. 3.1.1.3) was obtained after Superdex 200 gel filtration and Resource Q anionic exchange. The pure lipase, which is a glycosylated molecule, is a monomer having a molecular mass of about 44.8kDa, as determined by SDS-PAGE analysis. The lipase hydrolyses short and long-chain triacylglycerols and naphthol derivates at comparable rates. A specific activity of 1787U mg−1 and 475U mg−1 was measured with triolein and tributyrin as substrates, respectively, at pH 8.0 and 30°C in the absence of colipase. The lipase showed a Km, app of 3.22mM and kcat, app/Km, app of 0.303 × 103 mM−1 s−1 using triolein as substrate. Natural detergents, such as sodium deoxycholate, act as potent inhibitors of the lipase. This inhibition can be reversed by adding fresh oil emulsion. Result with tetrahydrolipstatin, an irreversible inhibitor, suggests that the lipase is a serine enzyme. Peptide sequences of the lipase were determined and compared with the full-length sequence of lipase which was obtained by the rapid amplification of cDNA ends method. The full cDNA of the pvl was 1,186bp, with a deduced protein of 362 amino acids that includes a consensus sequence (GXSXG) of the lipase superfamily of α/β-hydrolase. The gene exhibits features of conserved catalytic residues and high homology with various mammalian and insect lipase genes. A potential lid sequence is suggested for pvl. Springer Science+Business Media, LLC, 2010 Digestion nationallicence Lipase nationallicence Purification nationallicence Penaeus vannamei nationallicence Crustaceans nationallicence Rivera-Pérez Crisalejandra Centro de Investigaciones Biologicas del Noroeste (CIBNOR), Apdo. Postal 128, B.C.S. 23000, La Paz, Mexico aut García-Carreño Fernando Centro de Investigaciones Biologicas del Noroeste (CIBNOR), Apdo. Postal 128, B.C.S. 23000, La Paz, Mexico aut Saborowski Reinhard Alfred-Wegener-Institute für Polar- und Meeresforschung (AWI), Biologische Anstalt Helgoland, P.O. Box180, 27483, Helgoland, Germany aut Marine Biotechnology Springer-Verlag 13/2(2011-04-01), 284-295 1436-2228 13:2<284 2011 13 10126 https://doi.org/10.1007/s10126-010-9298-7 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10126-010-9298-7 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Rivera-Pérez Crisalejandra Centro de Investigaciones Biologicas del Noroeste (CIBNOR), Apdo. Postal 128, B.C.S. 23000, La Paz, Mexico aut NATIONALLICENCE 700 1- García-Carreño Fernando Centro de Investigaciones Biologicas del Noroeste (CIBNOR), Apdo. Postal 128, B.C.S. 23000, La Paz, Mexico aut NATIONALLICENCE 700 1- Saborowski Reinhard Alfred-Wegener-Institute für Polar- und Meeresforschung (AWI), Biologische Anstalt Helgoland, P.O. Box180, 27483, Helgoland, Germany aut NATIONALLICENCE 773 0- Marine Biotechnology Springer-Verlag 13/2(2011-04-01), 284-295 1436-2228 13:2<284 2011 13 10126 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer