Abstract
The expression and distribution of digestive cellulases along the midgut of Cerambyx cerdo larvae were analyzed for the first time and are presented in this article. Four groups of larvae were examined: larvae developed in the wild; larvae taken from the wild and successively reared on an artificial diet based on polenta; and larvae hatched in the laboratory and reared on two different artificial diets. Seven endocellulase and seven ?-D-glucosidase isoforms were detected in all midgut extracts of C. cerdo with a zymogram after native PAGE. We observed that C. cerdo larvae are capable of producing cellulase isoforms with different PAGE mobilities depending on the nutrient substrate. From our findings it can be assumed that, depending on the distribution of endocellulase and ?-D-glucosidase, cellulose molecules are first fragmented in the anterior and middle midgut by endo-?-1,4-glucanase; subsequently, the obtained fragments are broken down by ?-D-glucosidase mostly in middle midgut.
Highlights
Enzymatic activity against cellulose substrates was detected in the digestive tract of insects belonging to different insect orders (Martin, 1983; Watanabe and Tokuda, 2010, Oppert et al, 2010)
It has already been demonstrated that the expression of amylolytic and proteolytic enzymes depends on environmental conditions and nutrient substrate (Nenadović et al, 1994, Dojnov et al, 2010). The goal of this investigation was to examine the expression of endocellulase and β-D-glucosidase on the isoform level in larvae fed on two different artificial diets, and to detect these enzymes along the midgut of C. cerdo larval
Est in the crude midgut extract of C. cerdo larvae reared on oak sawdust (EOS) (Fig. 2a), while specific endocellulase activity (0.46 U/mg) was highest in the crude midgut extract of wild larvae (LW)
Summary
Enzymatic activity against cellulose substrates was detected in the digestive tract of insects belonging to different insect orders (Martin, 1983; Watanabe and Tokuda, 2010, Oppert et al, 2010). C. cerdo is listed as vulnerable on the IUCN Red list of threatened species (1996), in an Annex II Species of the EU Habitats and Species Directive (2009), and by the Decree on Protection of Natural Rarities of Serbia (1993) It is distributed in Europe, the Caucasus, Asia Minor and northern Africa (Kimoto and Duthie-Holt, 2006). It has already been demonstrated that the expression of amylolytic (on the isoenzyme level) and proteolytic enzymes depends on environmental conditions and nutrient substrate (Nenadović et al, 1994, Dojnov et al, 2010) The goal of this investigation was to examine the expression of endocellulase and β-D-glucosidase on the isoform level in larvae fed on two different artificial diets, and to detect these enzymes along the midgut of C. cerdo larval
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