Digestive proteolytic activity in Apodiphus amygdali Germar (Hemiptera: Pentatomidae): effect of endogenous inhibitors

• Main Authors: S. Ramzi, A. Zibaee
• Format: Article
• Language: English
• Published: PAGEPress Publications 2014-08-01
• Series: Journal of Entomological and Acarological Research
• Subjects: Apodiphus amygdali, digestive protease, inhibitor.
• Online Access: http://www.pagepressjournals.org/index.php/jear/article/view/1868
• ISSN: 2038-324X; 2279-7084

The digestive proteolytic profile of <em>Apodiphus amygdali</em> was determined by using several substrates and specific inhibitors. Analysis of optimal pH and temperature showed the highest enzymatic activity at the pH range of 6-7 and temperature of 40°C when azocasein was used as a substrate. By using a negative control, the presence of several specific proteases were determined including tryspin-like, chymotrypsinlike, elastase, cathepsin B, cathepsin L, amino- and carboxypeptidases in the midgut content of <em>A. amygdali</em>, with the highest and the lowest activities of cathepsin L and carboxypeptidase, respectively. pH dependency of specific proteases revealed optimal pHs of 9, 8 and 9 for trypsin-, chymotrypsin-like, 6 for cathepsins and 5-6 for carboxy- and aminopeptidases, respectively. Specific inhibitors, including phenylmethylsulfonyl fluoride, Na-p-tosyl-L-lysine chloromethyl ketone, Ntosyl- L-phenylalanine chloromethyl ketone, L-trans-epoxysuccinylleucylamido-(4-guanidino)-butane, phenanthroline and ethylendiamidetetraacetic acid, significantly decreased proteolytic activity, indicating the presence of different proteases in the midgut of <em>A. amygdali</em>. Extracted inhibitors from the midgut demonstrated significant inhibition of specific proteolytic activities of <em>A. amygdali</em> except for cathepsin B and aminopeptidase. The results indicated that determination of digestive proteolytic activity could be helpful to clarify digestion process in insects. Moreover, understanding the nature of digestive proteases might be used to develop several inhibitors for providing resistant crop varieties against pests.