Amylases from adults of Sitophilus oryzae (L.) and S. granarius (L.) were purified by using a sequential procedure of ammonium sulfate precipitation, glycogen-complex formation, and ion exchange chromatography. Amylase of S. oryaze was purified 47.4-fold to a specific activity of 478 unitdmg protein.One amylase unit equals 1 mg maltose hydrate producedlmin at 3OOC. Amylase of S. granarius was purified 85.4fold to a specific activity of 453 unitshg protein. Amylase of S. oryzae had a Km of 0.173% for soluble starch and consisted of two anionic isozyrnes with isoelectric points of pH 3.70 and pH 3.76. Amylase of S. granarius had a Km of 0.078% for starch and was a single protein with an isoelectric point of pH 3.76. Purified amylases of both species had molecular weights of 56,000 estimated by sodium dodecyl sulfatepolyacrylamide gel electrophoresis, were activated by chloride, and had double energies of activation calculated from Arrhenius plots. Based on fresh weights of adults feeding on whole wheat through 10 weeks of age, S. oryzae contained three-fold and eight-fold more amylase than S. granarius and S.zeamais Motschulsky, respectively. High amylase levels in S. oryzae may provide this species with an adaptive advantage when feeding on cereals containing naturally occurring amylase inhibitors.
In addition to typical columnar cells, dark-staining cells characterized by deep invaginations of basal plasma membranes were found throughout the midgut of adult Sitophilus granarius (L.). These invaginations formed intracellular channels that extended to the perinuclear region and indicated an involvement of these cells in secretion and (or) absorption. Cells with large vacuolelike structures that occasionally filled the entire supranuclear region were found in the anterior midgut, while multicellular crypts and cells that formed apical extrusions into the lumen were common in the posterior midgut. Fine structure of gastric caeca indicated functional differences between those located in anterior and posterior midgut regions. Numerous dark-staining granules were found in apical regions of cells of the anterior caeca, whereas elongated mitochondria were found in microvilli that made up the brush border of posterior caecal cells. Thus, although the midgut is not obviously differentiated into zones, there are different cell types in different regions of the gut involved in digestive and nutrient absorption processes. Evidence indicated that amylase in S. granarius is secreted by salivary glands whereas trypsin and aminopeptidase are secreted by midgut. A slow, continuous secretion of amylase occurs whereas proteinases are secreted in response to ingested food.
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