Eleven proteinase activity bands were detected in American cockroach (Periplaneta americana) gut. These were partially purified and characterized using a gel X-ray film contact print method. Cockroach gut proteinases (CGPs) show activity... more
Eleven proteinase activity bands were detected in American cockroach (Periplaneta americana) gut. These were partially purified and characterized using a gel X-ray film contact print method. Cockroach gut proteinases (CGPs) show activity over a broad range of pH with maximum activity between pH 6 and 10, and optimal activity at 50-70 degrees C. CGPs were partially purified by preparative gel electrophoresis and analyzed using synthetic substrates and inhibitors. Four of the proteases exhibited chymotrypsin-like (C1 to C4) activity and seven trypsin-like (T1 to T7) activity. Accuracy of the gel X-ray film contact print method is confirmed by including bovine chymotrypsin in CGP analysis. Inhibition of CGPs with different plant proteinaceous proteinase inhibitors allowed identification of potential CGP inhibitors. Our results imply that presence of several CGP activity bands, and their stability and activity over a broad pH and temperature range might contribute to adaptation of P. americana to extreme environmental conditions and the polyphagous nature of the species.
... lebbeck with inhibitory potential toward insect amylases Neeta D. Kalve • Purushottam R. Lomate • Vandana K. Hivrale ... Low molecular weight markers were purchased from Genei,Bangalore, India. All other chemi-cals used were of high... more
... lebbeck with inhibitory potential toward insect amylases Neeta D. Kalve • Purushottam R. Lomate • Vandana K. Hivrale ... Low molecular weight markers were purchased from Genei,Bangalore, India. All other chemi-cals used were of high analytical grade. ...
Eleven proteinase activity bands were detected in American cockroach (Periplaneta americana) gut. These were partially purified and characterized using a gel X-ray film contact print method. Cockroach gut proteinases (CGPs) show activity... more
Eleven proteinase activity bands were detected in American cockroach (Periplaneta americana) gut. These were partially purified and characterized using a gel X-ray film contact print method. Cockroach gut proteinases (CGPs) show activity over a broad range of pH with maximum activity between pH 6 and 10, and optimal activity at 50-70 degrees C. CGPs were partially purified by preparative gel electrophoresis and analyzed using synthetic substrates and inhibitors. Four of the proteases exhibited chymotrypsin-like (C1 to C4) activity and seven trypsin-like (T1 to T7) activity. Accuracy of the gel X-ray film contact print method is confirmed by including bovine chymotrypsin in CGP analysis. Inhibition of CGPs with different plant proteinaceous proteinase inhibitors allowed identification of potential CGP inhibitors. Our results imply that presence of several CGP activity bands, and their stability and activity over a broad pH and temperature range might contribute to adaptation of P. americana to extreme environmental conditions and the polyphagous nature of the species.
Proteinases from the hepatopancreas of freshwater crab Barytelphusa cunicularis were characterized with respect to their biochemical properties. Serine protease activities corresponding to trypsin and chymotrypsin were detected. A total... more
Proteinases from the hepatopancreas of freshwater crab Barytelphusa cunicularis were characterized with respect to their biochemical properties. Serine protease activities corresponding to trypsin and chymotrypsin were detected. A total of eight proteinase activity bands were detected in hepatopancreatic extract on X- ray film. Synthetic substrate and inhibitor studies revealed the presence of trypsin, chymotrypsin, and aminopeptidase like proteinases. Of total
ABSTRACT The angiotensin-converting enzyme (ACE) inhibitory activity of methanolic extract of Peganum Harmala seeds was studied. The active compound was isolated and identified as harmaline with spectroscopic data. In vitro ACE inhibition... more
ABSTRACT The angiotensin-converting enzyme (ACE) inhibitory activity of methanolic extract of Peganum Harmala seeds was studied. The active compound was isolated and identified as harmaline with spectroscopic data. In vitro ACE inhibition assay was performed using four concentrations of harmaline. Harmaline inhibited 48.7% ACE activity at 1 μg and was very close to the inhibition observed with the standard drug, captopril at 49.1% (1 μM). At 0.5 μg, 0.25 μg, and 0.1 μg, harmaline inhibited 42.5%, 32.3%, and 23.5% ACE activity, respectively. Results indicated that harmaline may act as a potential anti-hypertensive agent.
Starchy seeds are an important food and a source of dietary ingredients in many countries. However, they suffer from extensive predation by bruchids (weevils) and other pests. α-Amylase inhibitors are attractive candidates for the control... more
Starchy seeds are an important food and a source of dietary ingredients in many countries. However, they suffer from extensive predation by bruchids (weevils) and other pests. α-Amylase inhibitors are attractive candidates for the control of seed weevils, as these insects are highly dependent on starch as an energy source. A proteinaceous α-amylase inhibitor from the seeds of Achyranthes aspera was identified, purified and characterised. In electrophoretic analysis, two prominent amylase inhibitor activity bands (AI1 and AI2) were detected. The inhibitor was purified 9.99-fold with 1206.95 total amylase inhibitor units mg⁻¹ protein. The molecular weight of the purified inhibitor was around 6 kDa. The isolated α-amylase inhibitor was found to be resistant to heat and proteolysis. Feeding analysis of Callosobruchus maculatus larvae on a diet containing seed powder of A. aspera revealed that survival of the larvae was severely affected, with the highest mortality rate occurring on the fifth day of feeding. The isolated inhibitor inhibited the majority of amylase isoforms of C. maculatus, Tribolium confusum and Helicoverpa armigera in electrophoretic analysis and solution assays. The information obtained in the present investigation could be useful for a genetic engineering approach that would make seeds resistant to storage pest infestations.
Insect midgut proteases are known to be regulated by plant protease inhibitors. In the present study, the antinutritional effects of a variety of seed extracts against Periplaneta americana (Linnaeus) (Dictyoptera: Blattidae) midgut... more
Insect midgut proteases are known to be regulated by plant protease inhibitors. In the present study, the antinutritional effects of a variety of seed extracts against Periplaneta americana (Linnaeus) (Dictyoptera: Blattidae) midgut proteases are assessed in vitro and in vivo. ...
Wounding of plants by chewing insects or other damage induces the synthesis of defensive proteinase inhibitors (PI) in both wounded and distal unwounded leaves. In the present paper we report the characterization of inducible defensive PI... more
Wounding of plants by chewing insects or other damage induces the synthesis of defensive proteinase inhibitors (PI) in both wounded and distal unwounded leaves. In the present paper we report the characterization of inducible defensive PI from pigeon pea (Cajanus cajan) and its in vitro interaction with Helicoverpa armigera gut proteinases (HGP). We found that PI activity was induced in local as well as systemic leaves of pigeon pea by the wounding and methyl jasmonate (MeJA) application. Consistent induction of PI was observed in two wild cultivars of pigeon pea at various growth stages. The estimated molecular weight of inducible PI was ~16.5 kDa. Electrophoretic analysis and enzyme assays revealed that the induced PI significantly inhibited total gut proteinase as well as trypsin-like activity from the midgut of H. armigera. The induced PI was found to be inhibitor of trypsin as well as chymotrypsin. Study could be important to know the further roles of defensive PIs.