Fractionation of digestive proteinases from Tenebrio molitor (Coleoptera: Tenebrionidae) larvae and role in protein digestion.
Tenebrio molitor larval digestive proteinases were purified and characterized by gel filtration chromatography combined with activity electrophoresis. Cysteine proteinases, consisting of at least six distinct activities, were found in three chromatographic peaks in anterior and posterior midgut chromatographies. The major activity in the anterior midgut, peak cys II, consisted of cysteine proteinases with Mm of 23 kDa. The predominant peak in the posterior, cys I, was represented by 38 kDa proteinases. The activities of all cysteine proteinases were maximal in buffers from pH 5.0 to 7.0, with 80% stability at pH values from 4.0 to 7.0. In the conditions of the last third of the midgut, the activity and stability of cysteine proteinases was sharply decreased. Trypsin-like activity included a minor peak of "heavy" trypsins with Mm 59 kDa, located mainly in the anterior midgut. An in vitro study of the initial stages of digestion of the main dietary protein, oat 12S globulin, by anterior midgut proteinases revealed that hydrolysis occurred through the formation of intermediate high-Mm products, similar to those formed during oat seed germination. Cysteine proteinases from the cys III peak and heavy trypsins were capable of only limited proteolysis of the protein, whereas incubation with cys II proteinases resulted in substantial hydrolysis of the globulin.
Vinokurov
K S
KS
Department of Entomology, Biological Faculty, Moscow State University, Moscow 119992, Russia.
Elpidina
E N
EN
Oppert
B
B
Prabhakar
S
S
Zhuzhikov
D P
DP
Dunaevsky
Y E
YE
Belozersky
M A
MA
D000818:Animals; D018554:Avena / Q000378:metabolism; D003546:Cysteine Endopeptidases / Q000302:isolation & purification* / Q000378:metabolism* / Q000502:physiology; D004063:Digestion / Q000502:physiology; D004064:Digestive System / Q000201:enzymology / Q000378:metabolism; D006868:Hydrolysis; D019476:Insect Proteins / Q000302:isolation & purification* / Q000378:metabolism* / Q000502:physiology; D007814:Larva / Q000201:enzymology; D014674:Plant Proteins, Dietary / Q000378:metabolism; D013712:Tenebrio / Q000201:enzymology* / Q000254:growth & development*
D019476:Insect Proteins; D014674:Plant Proteins, Dietary; D003546:Cysteine Endopeptidases