Purification, partial characterization and role in lipid transport to developing oocytes of a novel lipophorin from the cowpea weevil, Callosobruchus maculatus

Lipid transport in arthropods is achieved by highly specialized lipoproteins, which resemble those described in vertebrate blood. Here we describe purification and characterization of the lipid-apolipoprotein complex, lipophorin (Lp), from adults and larvae of the cowpea weevil Callosobruchus maculatus. We also describe the Lp-mediated lipid transfer to developing oocytes. Lps were isolated from homogenates of C. maculatus larvae and adults by potassio bromide gradient and characterized with respect to physicochemical properties and lipid content. The weevil Lp (465 kDa) and larval Lp (585 kDa), with hydrated densities of 1.22 and 1.14 g/mL, contained 34 and 56 percent lipids and 9 and 7 percent carbohydrates, respectively. In both Lps, mannose was the predominant monosaccharide detected by paper chromatography. SDS-PAGE revealed two apolipoproteins in each Lp with molecular masses of 225 kDa (apolipoprotein-I) and 79 kDa (apolipoprotein-II). The lipids were extracted and analyzed by thin-layer chromatography. The major phospholipids found were phosphatidylserine, phosphatidylcholine and phosphatidylethanolamine in adult Lp, and phosphatidylcholine, phosphatidylethanolamine and sphingomyelin in larval Lp. Hydrocarbons, fatty acids and triacylglycerol were the major neutral lipids found in both Lps. Lps labeled in the protein moiety with radioactive iodine (125I-iodine) or in the lipid moiety with fluorescent lipids revealed direct evidence of endocytic uptake of Lps in live oocytes of C. maculatus.

Efecto de la extrusión sobre la biodisponibilidad de proteína y almidón en mezclas de harinas de maíz y frijol lima / Effect of extrusion on protein and starch bioavailability in corn and lima bean flour blends

La extrusión se emplea para producir expandidos (botanas o snacks), crujientes y quebradizos. El impacto nutricional de este tipo de proceso no ha sido suficientemente estudiado, por ello se evaluó la biodisponibilidad “in vitro” e “in vivo” de la proteína y el almidón en mezclas de harinas de maíz (Zea mays) (M) y de frijol lima (Phaseolus lunatus) (F), tanto crudas como extrudidas. Las mezclas de harinas 75M/25F y 50M/50F (p/p) y fueron procesadas en un extrusor Brabender a 160°C, 100 rpm, y 15,5% de humedad. La composición proximal indicó un aumento del tenor de proteína y cenizas y una disminución de la grasa en los productos extrudidos. La digestibilidad in vitro de la proteína fue mayor en los extrudidos (82%) que en las harinas crudas (77%). Los contenidos de almidón potencialmente disponible y almidón resistente total disminuyeron con la extrusión. Los ensayos in vitro indican que la extrusión mejora la digestibilidad de la proteína y el almidón en las mezclas estudiadas. La biodisponibilidad in vivo se evaluó utilizando gorgojos de arroz (Sithophilus oryzae) como modelo biológico. Los biomarcadores más descriptivos de los cambios sugeridos por las pruebas in vivo fueron: el contenido de proteínas corporales, que incrementó por efecto de la extrusión, y la actividad de la alfa-amilasa intestinal, que disminuyó a consecuencia del procesamiento. Se concluye que la extrusión mejora apreciablemente la calidad nutricional de mezclas de maíz y frijol de lima.

Effect of extrusion on protein and starch bioavailability in corn and lima bean flour blends. Extrusion is used to produce crunchy expanded foods, such as snacks. The nutritional impact of this process has not been studied sufficiently. In this study, in vitro and in vivo protein and starch bioavailability was evaluated in both raw and extruded corn (Zea mays)(C) and lima bean (Phaseolus lunatus)(B) flour blends, prepared in 75C/25B and 50C/ 50B (p/p) proportions. These were processed with a Brabender extruder at 160°C, 100 rpm and 15.5% moisture content. Proximate composition showed that in the extruded products protein and ash contents increased whereas the fat level decreased. In vitro protein digestibility was higher in the extrudates (82%) than in the raw flours (77%). Potentially available starch and resistant starch contents decreased with extrusion. The in vitro assays indicated that extrusion improved protein and starch availability in the studied blends. In vivo bioavailability was evaluated using the rice weevil (Sithophilus oryzae) as a biological model. The most descriptive biomarkers of the changes suggested by the in vivo tests were body protein content (increased by extrusion) and intestinal alpha-amylase activity (decreased by processing). Overall, results suggest that extrusion notably increases the nutritional quality of corn and lima bean flour blends.