In Vivo Bioinsecticidal Activity toward Ceratitis capitata (Fruit Fly) and Callosobruchus maculatus (Cowpea Weevil) and in Vitro Bioinsecticidal Activity toward Different Orders of Insect Pests of a Trypsin Inhibitor Purified from Tamarind Tree (Tamarindus indica) Seeds

Abstract

A proteinaceous inhibitor with high activity against trypsin-like serine proteinases was purified from seeds of the tamarind tree (Tamarindus indica) by gel filtration on Shephacryl S-200 followed by a reverse-phase HPLC Vidac C18 TP. The inhibitor, called the tamarind trypsin inhibitor (TTI), showed a Mr of 21.42 kDa by mass spectrometry analysis. TTI was a noncompetitive inhibitor with a Ki value of 1.7 x 10(-9) M. In vitro bioinsecticidal activity against insect digestive enzymes from different orders showed that TTI had remarkable activity against enzymes from coleopteran, Anthonomus grandis (29.6%), Zabrotes subfasciatus (51.6%), Callosobruchus maculatus (86.7%), Rhyzopertha dominica(88.2%), and lepidopteron, Plodia interpuncptella (26.7%), Alabama argillacea (53.8%), and Spodoptera frugiperda (75.5%). Also, digestive enzymes from Diptera, Ceratitis capitata (fruit fly), were inhibited (52.9%). In vivo bioinsecticidal assays toward C. capitata and C. maculatus larvae were developed. The concentration of TTI (w/w) in the artificial seed necessary to cause 50% mortality (LD50) of larvae was 3.6%, and that to reduce mass larvae by 50.0% (ED50) was 3.2%. Furthermore, the mass C. capitata larvae were affected at 53.2% and produced approximately 34% mortality at a level of 4.0% (w/w) of TTI incorporated in artificial diets.

Effects of gamma‐irradiation on midgut proteolytic activity of the mediterranean fruit fly, Ceratitis capitata (Diptera: Tephritidae)

“…We have found that larvae of C. capitata presented the same proteolytic profile as adults, since their digestive system is based on cathepsin-D-, trypsin-, chymotrypsin-, leucine aminopeptidase-, and carboxypeptidase A-and B-like activities. Our results confirm the presence of both alkaline and acid proteases identified in the larval midgut of this species (Rabossi et al, 2000;Araujo et al, 2005;Silva et al, 2006). However, since the pH requirements of these two types of proteases are quite different, further work will be necessary to determine if their spatial organization within the midgut accommodates conditions suitable for maximal activity.…”

“…Silva et al (2006) reported that larvae and adult C. capitata rely on serine proteases for protein digestion, with a small contribution of aspartic and cysteine digestive proteases. However, apart from the trypsin-and chymotrypsin-like protease activities identified (Araujo et al, 2005;Silva et al, 2006), the nature of other digestive proteases in this species is largely unknown. Histopathological evidence has showed that sterile males of C. capitata sustain extreme radiation damage to their intestinal epithelial tissue, showing signs of necrosis along with apoptosis and nuclear damage (Niyazi et al, 2004).…”

Effect of trypsin inhibitor from Crotalaria pallida seeds on Callosobruchus maculatus (cowpea weevil) and Ceratitis capitata (fruit fly)

“…The role of these inhibitors as defensive compounds against predators was studied as early as 1947 when Mickel and Standish [36] observed that larvae of certain insects were unable to develop on soybean products. A later report showed that trypsin inhibitors were toxic to Tribolium confusum (flour beetle) larvae [37].…”

Purification, characterization and evaluation of insecticidal activity of trypsin inhibitor from Albizia lebbeck seeds

“…Earlier trypsin inhibitors with low value of K i were reported from Poecilanthe parviflora (Garcia et al 2004), Calliandra selloi Macbride (Yoshizaki et al 2007), Albizia kalkora (Zhou et al 2008) and Cassia obtusifolia (Liao et al 2007). The analysis of Lineweaver-Burk plot showed the inhibitor to be of noncompetitive type like trypsin inhibitors from Adenanthera pavonina , Vicia faba (Gupta et al 2000), and Tamarindus indica (Araujo et al 2005). Shee and Sharma (2007), Oliveira et al (2007) and Bhattacharyya et al (2006), however found the inhibitors to be of competitive type.…”