Pancreatic Trypsin Research Articles

INHIBITORY EFFECT OF CARBOHYDRAZ ON SECRETORY ACTIVITY OF THE PANCREAS

Since the time of Ivan Petrovich Pavlov's work, it is known that the juice collected from the pancreatic fistula, when it is re-introduced into the duodenum, inhibits the secretion of the pancreas. Most recently, in the works of American, French and Japanese physiologists, this inhibition was associated with the fact that trypsinogen secreted in the pancreatic secretion and trypsin contained in the duodenal contents inhibit the release of the hormone cholicystokininpancreosimin by duodenal endocrine cells - a powerful stimulator of enzyme secretion by the pancreas. This explanation of the reverse reproduction of pancreatic secretion from the duodenum is recognized by most experts. In connection with the above, the study of this problem seems to be an urgent task.

Coexistence of acute pancreatitis and an obstructing upper ureteral calculus: does a calculus cause pancreatitis? a case report.

A 36-year-old male patient presented with epigastric, and right loin pain with decreased urine output for 3 days. On physical examination, he had central abdominal and right flank tenderness. An abdominal ultrasound showed mild to moderate ascites, a hyperechoic pancreas, a small (26×77mm) left kidney with increased echogenicity, right renal hypertrophy with moderate hydronephrosis, and a dilated upper ureter due to a 10mm obstructing stone with a perirenal fluid collection. The diagnosis of acute pancreatitis with an obstructing right upper ureteric stone was established. Under spinal anesthesia, an emergency ureteroscopy with laser fragmentation of the stone was performed, and a JJ stent was inserted. He developed postobstructive diuresis and his renal function was improved with a rapid decline of pancreatic enzymes as well. Two theories explain the presentation of acute pancreatitis by ureteral obstruction. First, the obstructed severe hydronephrotic kidney compresses the duodenum and head of the pancreas, obstructing the distal part of the common bile duct and triggering the elevation of pancreatic duct pressure, bile reflux, trypsin activation, and pancreatic autodigestion. The second theory states that acute pancreatitis develops when urine is extravasated from an obstructed kidney into the adjacent tissues, irritating the uncinate process of the pancreas. Although mentioning ureteral obstruction as a cause of pancreatitis is scarce, the clinician should be aware that in each case of ureteral obstruction, the emergence of acute pancreatitis is a possible complication.

Probing protein stability: towards a computational atomistic, reliable, affordable, and improvable model.

We present an improved application of a recently proposed computational method designed to evaluate the change of free energy as a function of the average value of a suitably chosen collective variable in proteins. The method is based on a full atomistic description of the protein and its environment. The goal is to understand how the protein melting temperature changes upon single-point mutations, because the sign of the temperature variation will allow us to discriminate stabilizing vs. destabilizing mutations in protein sequences. In this refined application the method is based on altruistic well-tempered metadynamics, a variant of multiple-walkers metadynamics. The resulting metastatistics is then modulated by the maximal constrained entropy principle. The latter turns out to be especially helpful in free-energy calculations as it is able to alleviate the severe limitations of metadynamics in properly sampling folded and unfolded configurations. In this work we apply the computational strategy outlined above in the case of the bovine pancreatic trypsin inhibitor, a well-studied small protein, which is a reference for computer simulations since decades. We compute the variation of the melting temperature characterizing the folding-unfolding process between the wild-type protein and two of its single-point mutations that are seen to have opposite effect on the free energy changes. The same approach is used for free energy difference calculations between a truncated form of frataxin and a set of five of its variants. Simulation data are compared to in vitro experiments. In all cases the sign of the change of melting temperature is reproduced, under the further approximation of using an empirical effective mean-field to average out protein-solvent interactions.

Study on the mechanism of early pancreatic exocrine function changes in severely scalded rats

Study on the mechanism of early pancreatic exocrine function changes in severely scalded rats

Accounting for Solvation Correlation Effects on the Thermodynamics of Water Networks in Protein Cavities

Macromolecular recognition and ligand binding are at the core of biological function and drug discovery efforts. Water molecules play a significant role in mediating the protein-ligand interaction, acting as more than just the surrounding medium by affecting the thermodynamics and thus the outcome of the binding process. As individual water contributions are impossible to measure experimentally, a range of computational methods have emerged to identify hydration sites in protein pockets and characterize their energetic contributions for drug discovery applications. Even though several methods model solvation effects explicitly, they focus on determining the stability of specific water sites independently and neglect solvation correlation effects upon replacement of clusters of water molecules, which typically happens in hit-to-lead optimization. In this work, we rigorously determine the conjoint effects of replacing all combinations of water molecules in protein binding pockets through the use of the RE-EDS multistate free-energy method, which combines Hamiltonian replica exchange (RE) and enveloping distribution sampling (EDS). Applications on the small bovine pancreatic trypsin inhibitor and four proteins of the bromodomain family illustrate the extent of solvation correlation effects on water thermodynamics, with the favorability of replacement of the water sites by pharmacophore probes highly dependent on the composition of the water network and the pocket environment. Given the ubiquity of water networks in biologically relevant protein targets, we believe our approach can be helpful for computer-aided drug discovery by providing a pocket-specific and a priori systematic consideration of solvation effects on ligand binding and selectivity.

Correlation between protein conformations and water structure and thermodynamics at high pressure: A molecular dynamics study of the Bovine Pancreatic Trypsin Inhibitor (BPTI) protein.

Pressure-induced perturbation of a protein structure leading to its folding-unfolding mechanism is an important yet not fully understood phenomenon. The key point here is the role of water and its coupling with protein conformations as a function of pressure. In the current work, using extensive molecular dynamics simulation at 298K, we systematically examine the coupling between protein conformations and water structures of pressures of 0.001, 5, 10, 15, 20 kbar, starting from (partially) unfolded structures of the protein Bovine Pancreatic Trypsin Inhibitor (BPTI). We also calculate localized thermodynamics at those pressures as a function of protein-water distance. Our findings show that both protein-specific and generic effects of pressure are operating. In particular, we found that (1) the amount of increase in water density near the protein depends on the protein structural heterogeneity; (2) the intra-protein hydrogen bond decreases with pressure, while the water-water hydrogen bond per water in the first solvation shell (FSS) increases; protein-water hydrogen bonds also found to increase with pressure, (3) with pressure hydrogen bonds of waters in the FSS getting twisted; and (4) water's tetrahedrality in the FSS decreases with pressure, but it is dependent on the local environment. Thermodynamically, at higher pressure, the structural perturbation of BPTI is due to the pressure-volume work, while the entropy decreases with the increase of pressure due to the higher translational and rotational rigidity of waters in the FSS. The local and subtle effects of pressure, found in this work, are likely to be typical of pressure-induced protein structure perturbation.

A case of umbilical cord hemorrhagic ulceration with congenital upper gastrointestinal atresia

A 23-year-old woman (gravida 3, para 2) was referred to our hospital at 29 weeks of gestation by a nearby doctor due to hydramnios and suspected umbilical cord hernia. Ultrasound and magnetic resonance imaging indicated hydramnios, umbilical cord hernia, and a dilated stomach, with suspicion of upper gastrointestinal atresia. She was hospitalized at 33 weeks of gestation owing to membrane rupture. Fetal bradycardia was observed during delivery, and an emergency cesarean section was performed. A female infant weighing 1,799 g was delivered (Apgar score: 1/1); hemorrhagic ulceration of the umbilical cord was observed. Pancreatic phospholipase A2, trypsin, lipase, and bile acid levels in the amniotic fluid were elevated. The infant died of hemorrhagic shock and circulatory failure on the second day after birth. Based on these findings, we hypothesize that umbilical cord ulceration or hemorrhage may occur in congenital upper gastrointestinal atresia due to discharge of fetal gastrointestinal enzymes into the amniotic fluid.

Overt Vitamin B-6 Deficiency Affects Rat Pancreatic Digestive Enzyme and Glutathione Reductase Activities

Previous reports suggest that vitamin B-6 deficiency contributes to pancreatic insufficiency. However, the susceptibility of pancreatic function to marginal vitamin B-6 intake has not been defined. The present study examines digestive enzyme activity and steady-state mRNA levels, as well as antioxidant enzyme status from rats fed different vitamin B-6 diets. Groups (n = 12) of adult female Long-Evans rats were assigned to five dietary groups and fed their respective diets for 6 wk. Control and food-restricted rats were fed the control diet (7 mg pyridoxine/kg diet) freely, or food intake was restricted to the lowest intake of the experimental groups. The experimental groups were fed purified diets containing 0 (deficient), 0.25 or 1 (marginal) mg pyridoxine/kg diet. Plasma amylase and pancreatic amylase, trypsin and chymotrypsin activities were significantly lower in deficient rats compared with rats fed the control diet. Lower enzyme activities were accompanied by 83 and 55% lower amylase and trypsinogen mRNA levels compared with levels in rats fed the control diet. Other than low glutathione reductase in deficient rats, pancreatic antioxidant enzyme activity was similar in all dietary groups. These data suggest that the exocrine pancreas is impaired by vitamin B-6 deficiency, but marginal pyridoxine intake maintains function.

Oxidative Protein Folding Promotion by Imidazoyl-conjugated Thiol

The promotion of protein folding is an important subject that can contribute to the efficient production of biological medicines. Here we report the promotion of disulfide bond-coupled protein folding by imidazoyl-conjugated thiol (ImdSH). ImdSH accelerated the disulfide bond formation and folding of reduced ribonuclease A and bovine pancreatic trypsin inhibitor relative to glutathione as a conventionally used additive.

Tragacanth Gum and Linseed Gum as Adhesives Improved the Survival, Digestive Function, Antioxidant Enzyme Activities, and Immunity in Large Yellow Croaker (Larimichthys crocea) Larvae

A 30-day feeding experiment was designed to investigate the effects of different adhesives on survival, growth performance, digestive enzyme activities, antioxidant capacity, and inflammation response of large yellow croaker (Larimichthys crocea) larvae (initial weight 8.40 ± 0.18 mg). Four iso-nitrogenous (55% crude protein) and iso-lipidic (22% crude lipid) microbound diets (MBD) were formulated to contain 2% sodium alginate (SA), 2% tragacanth gum (TG), 2% linseed gum (LG), and 2% poloxamer (PX), respectively. Results showed that larvae fed with TG and LG diets had significantly higher survival rate than the larvae fed with SA and PX ( P < 0.05 ). Larvae fed with the PX diet had lower final weight and specific growth rate than the other groups ( P < 0.05 ). Larvae fed with TG diet had significant higher activities of lipase in pancreatic segments and trypsin in intestinal segments than the larvae fed with the PX ( P < 0.05 ). Meanwhile, the lipase activities of larvae fed with LG was significantly higher in intestinal segments than those fed with SA and PX diets ( P < 0.05 ). Larvae fed with TG and LG diets had significantly higher activities of AKP in brush border membranes than larvae fed with SA and PX diets ( P < 0.05 ). Moreover, the mRNA expression of pcna and zo-1 increased, respectively, in larvae fed TG and LG diets compared with the other groups ( P < 0.05 ). Larvae fed with LG and TG diets had significantly higher activities of SOD and T-AOC than those fed with PX diet ( P < 0.05 ). The content of GSH increased significantly in larvae fed with SA, TG, and LG diets compared with PX treatment ( P < 0.05 ). The activities of iNOS significantly increased in larvae fed with TG and LG diets compared to other groups ( P < 0.05 ). Meanwhile, the mRNA expression of il-10 significantly increased in TG and LG groups ( P < 0.05 ). In conclusion, results of the study demonstrated that TG and LG can be used as feed binders for microdiets with positive effects on survival, activities of digestive enzymes, antioxidant capacity, and inflammatory response. However, PX inhibited the survival and growth, and further study is needed to determine the suitable usage and dosage.

Pancreatic Secretory Trypsin Inhibitor (SPINK1) Gene Mutation in Patients with Acute Alcohol Pancreatitis (AAP) Compared to Healthy Controls and Heavy Alcohol Users without Pancreatitis

Only 3-5% of heavy alcohol users develop acute alcohol pancreatitis (AAP). This suggests that additional triggers are required to initiate the inflammatory process. Genetic susceptibility contributes to the development of AAP, and SPINK1 mutation is a documented risk factor. We investigated the prevalence of the SPINK1(N34S) mutation in patients with AAP compared to heavy alcohol users who had never suffered an episode of pancreatitis. Blood samples for the mutational analysis from patients with first episode (n = 60) and recurrent AAP (n = 43) and from heavy alcohol users without a history of AAP (n = 98) as well as from a control population (n = 1914) were obtained. SPINK1 mutation was found in 8.7% of the patients with AAP. The prevalence was significantly lower in healthy controls (3.4%, OR 2.72; 1.32-5.64) and very low in alcoholics without pancreatitis (1.0%, OR 9.29; 1.15-74.74). In a comparison adjusted for potential cofounders between AAP patients and alcoholics, SPINK1 was found to be an independent marker for AAP. The prevalence of the SPINK1 mutation is overrepresented in AAP patients and very low in alcoholics without pancreatitis. This finding may play a role in understanding the variable susceptibility to AAP found in heavy alcohol users.

The Non-native Disulfide-Bond-Containing Landscape Orthogonal to the Oxidative Protein-Folding Trajectory: A Necessary Evil?

Oxidative protein folding describes the process by which disulfide-bond-containing proteins mature from their ribosomal, fully reduced and unfolded, origins. Over the past 40 years, a number of exemplar proteins including bovine pancreatic ribonuclease A (RNaseA), bovine pancreatic trypsin inhibitor (BPTI), and hen egg-white lysozyme (HEWL), among others, have provided rich insight into the nature of the intermolecular interactions that drive the formation of the native, biologically active fold. In this Review Article, we revisit the oxidative folding process of RNaseA with a focus on reconciling the role of non-native disulfide-bond-containing species that populate the oxidative folding landscape. Toward gaining such an understanding, we project the regeneration pathway onto a Cartesian coordinate system. This helps not only to recognize the magnitude of the seemingly "fruitless", non-native disulfide-bond-containing species that lie orthogonal to the "native-protein-forming" reaction progress but also to reconcile a role for their existence in the regenerative trajectory. Finally, we superimpose the folding funnel onto the regeneration trajectory to draw parallels between oxidative folders and conformational folders (proteins that lack disulfide bonds). The overall objective is to provide the reader with a semi-quantitative description of oxidative protein folding and the barriers to successful regeneration while underscoring a role of seemingly fruitless intermediates.

Protein 3D Hydration: A Case of Bovine Pancreatic Trypsin Inhibitor

Characterization of the hydrated state of a protein is crucial for understanding its structural stability and function. In the present study, we have investigated the 3D hydration structure of the protein BPTI (bovine pancreatic trypsin inhibitor) by molecular dynamics (MD) and the integral equation method in the three-dimensional reference interaction site model (3D-RISM) approach. Both methods have found a well-defined hydration layer around the protein and revealed the localization of BPTI buried water molecules corresponding to the X-ray crystallography data. Moreover, under 3D-RISM calculations, the obtained positions of waters bound firmly to the BPTI sites are in reasonable agreement with the experimental results mentioned above for the BPTI crystal form. The analysis of the 3D hydration structure (thickness of hydration shell and hydration numbers) was performed for the entire protein and its polar and non-polar parts using various cut-off distances taken from the literature as well as by a straightforward procedure proposed here for determining the thickness of the hydration layer. Using the thickness of the hydration shell from this procedure allows for calculating the total hydration number of biomolecules properly under both methods. Following this approach, we have obtained the thickness of the BPTI hydration layer of 3.6 Å with 369 water molecules in the case of MD simulation and 3.9 Å with 333 water molecules in the case of the 3D-RISM approach. The above procedure was also applied for a more detailed description of the BPTI hydration structure near the polar charged and uncharged radicals as well as non-polar radicals. The results presented for the BPTI as an example bring new knowledge to the understanding of protein hydration.

Water Network in the Binding Pocket of Fluorinated BPTI-Trypsin Complexes─Insights from Simulation and Experiment.

Structural waters in the S1 binding pocket of β-trypsin are critical for the stabilization of the complex of β-trypsin with its inhibitor bovine pancreatic trypsin inhibitor (BPTI). The inhibitor strength of BPTI can be modulated by replacing the critical lysine residue at the P1 position by non-natural amino acids. We study BPTI variants in which the critical Lys15 in BPTI has been replaced by α-aminobutyric acid (Abu) and its fluorinated derivatives monofluoroethylglycine (MfeGly), difluoroethylglycine (DfeGly), and trifluoroethylglycine (TfeGly). We investigate the hypothesis that additional water molecules in the binding pocket can form specific noncovalent interactions with the fluorinated side chains and thereby act as an extension of the inhibitors. We report potentials of mean force (PMF) of the unbinding process for all four complexes and enzyme activity inhibition assays. Additionally, we report the protein crystal structure of the Lys15MfeGly-BPTI-β-trypsin complex (pdb: 7PH1). Both experimental and computational data show a stepwise increase in inhibitor strength with increasing fluorination of the Abu side chain. The PMF additionally shows a minimum for the encounter complex and an intermediate state just before the bound state. In the bound state, the computational analysis of the structure and dynamics of the water molecules in the S1 pocket shows a highly dynamic network of water molecules that does not indicate a rigidification or stabilizing trend in regard to energetic properties that could explain the increase in inhibitor strength. The analysis of the energy and the entropy of the water molecules in the S1 binding pocket using grid inhomogeneous solvation theory confirms this result. Overall, fluorination systematically changes the binding affinity, but the effect cannot be explained by a persistent water network in the binding pocket. Other effects, such as the hydrophobicity of fluorinated amino acids and the stability of the encounter complex as well as the additional minimum in the potential of mean force in the bound state, likely influence the affinity more directly.

Effects of Proteases from Pineapple and Papaya on Protein Digestive Capacity and Gut Microbiota in Healthy C57BL/6 Mice and Dose-Manner Response on Mucosal Permeability in Human Reconstructed Intestinal 3D Tissue Model.

Cysteine proteases obtained from the stem of pineapple or papaya latex, bromelain and papain, respectively, exhibit a broad spectrum of beneficial effects on human health. However, their effects on gut microbiota composition or dose-manner effects on the intestinal integrity of healthy tissue have not been evaluated. In this study, C57BL/6 young, healthy mice were fed bromelain or papain in a dose of 1 mg per animal/day for three consecutive days, followed by the assessment of digestive protein capacity, intestinal morphology and gut microbiota composition. Furthermore, a human reconstructed 3D tissue model EpiIntestinal (SMI-100) was used to study the effects of 1, 0.1 and 10 mg/mL doses of each enzyme on tissue integrity and mucosal permeability using TEER measurements and passage of Lucifer Yellow marker from the apical to the basolateral side of the mucosa. The results indicated that fruit proteases have the potential to modulate gut microbiota with decreasing abundance of Proteobacteria and increasing beneficial Akkermansia muciniphila. The enhancement of pancreatic trypsin was observed in bromelain and papain supplementation, while bromelain also increased the thickness of the ileal mucosa. Furthermore, an in vitro study showed a dose-dependent interruption in epithelial integrity, which resulted in increased paracellular permeability by the highest doses of enzymes. These findings define bromelain and papain as promising enzymatic supplementation for controlled enhancement of paracellular uptake when needed, together with beneficial effects on the gut microbiota.

The emergence of protein dynamics simulations: how computational statistical mechanics met biochemistry

In this essay, we aim to illustrate how Martin Karplus and his research group effectively set in motion the engine of molecular dynamics (MD) simulations of biomolecules. This process saw its prodromes between 1969 and the early 1970s with Karplus’ landing in biology, a transition that came to fruition with the treatment of 11-cis-retinal photoisomerization and the development of an allosteric model to account for the mechanism of cooperativity in hemoglobin. In 1977, J. Andrew McCammon, Bruce Gelin, and Martin Karplus published an article in Nature reporting the MD simulation of bovine pancreatic trypsin inhibitor (BPTI). This publication helped initiate the merger of computational statistical mechanics and biochemistry, a process that Karplus undertook at a later stage and whose beginnings we propose to reconstruct in this article through unpublished accounts of the key people who participated in this endeavor.

Formation of structured clots, gastric emptying and hydrolysis kinetics of yak milk during in vitro dynamic gastrointestinal digestion: Impact of different heat treatments

Yak milk is highly nutritious in terms of abundant protein and fat, whereas little is known regarding the influence and mechanism of heat treatments on its gastric emptying and hydrolysis kinetics during digestion in the human gastrointestinal tract. In this study, a bionic human stomach-intestine system was employed to examine in vitro dynamic gastrointestinal digestion of fresh raw and heated yak milks. During gastric digestion, the formation of protein clots or aggregates was observed for all the milk samples. However, significantly more clots were found from autoclaved milk and raw milk, with a dry weight of 14.1 and 10.9 g from 200 g initial milk sample after 30 min digestion, respectively, compared to pasteurized (6.9 g) and microwaved (9.2 g) milks. For both raw and heated milks, the large-sized coalesced fat globules formed in the stomach were rapidly dissociated to small and uneven droplets upon digestion in the small intestinal conditions due to the highly efficient hydrolysis of milk fat globule membrane (MFGM) proteins and fat globules by pancreatic trypsin and lipase. Due to the formation of more dense-structured clots, a longer gastric emptying half-time was shown for the autoclaved (41.1 min) and raw (38.4 min) milks in comparison to that for the yak milks treated with microwave (34.2 min) and pasteurization (36.2 min). Consistently, the pasteurized milk (92.5 %) exhibited the highest protein digestibility at the end of digestion, followed by microwaved milk (87.8 %) ≈ autoclaved milk (86.1 %) > raw milk (80.8 %). The amount of free fatty acids (FFAs) released during digestion in general followed a similar order with that for proteolysis among the milk samples. This study highlights the crucial role of the formation of structured clots in the gastric emptying and hydrolysis of proteins and fat globules. This information will provide a mechanistic understanding of digestion kinetics of yak milk as impacted by heat treatments.

Insight into the structural basis of the dual inhibitory mode of Lima bean (Phaseolus lunatus) serine protease inhibitor.

Bovine pancreatic trypsin was crystallized, in-complex with Lima bean trypsin inhibitor (LBTI) (Phaseolus lunatus L.), in the form of a ternary complex. LBTI is a Bowman-Birk-type bifunctional serine protease inhibitor, which has two independent inhibitory loops. Both of the loops can inhibit trypsin, however, only the hydrophobic loop is specific for inhibiting chymotrypsin. The structure of trypsin incomplex with the LBTI has been solved and refined at 2.25 Å resolution, in the space group P41, with Rwork /Rfree values of 18.1/23.3. The two binding sites of LBTI differ in only two amino acids. Lysine and leucine are the key residues of the two different binding loops positioned at the P1, and involved in binding the S1 binding site of trypsin. The asymmetric unit cell contains two molecules of trypsin and one molecule of LBTI. The key interactions include hydrogen bonds between LBTI and active site residues of trypsin. The 3D structure of the enzyme-inhibitor complex provided details insight into the trypsin inhibition by LBTI. To the best of our knowledge, this is the first report on the structure of trypsin incomplex with LBTI.

Characterizing Metastable States with the Help of Machine Learning

Present-day atomistic simulations generate long trajectories of ever more complex systems. Analyzing these data, discovering metastable states, and uncovering their nature are becoming increasingly challenging. In this paper, we first use the variational approach to conformation dynamics to discover the slowest dynamical modes of the simulations. This allows the different metastable states of the system to be located and organized hierarchically. The physical descriptors that characterize metastable states are discovered by means of a machine learning method. We show in the cases of two proteins, chignolin and bovine pancreatic trypsin inhibitor, how such analysis can be effortlessly performed in a matter of seconds. Another strength of our approach is that it can be applied to the analysis of both unbiased and biased simulations.

Bovine pancreatic trypsin inhibitor and soybean Kunitz trypsin inhibitor: Differential effects on proteases and larval development of the soybean pest Anticarsia gemmatalis (Lepidoptera: Noctuidae)

Pest management is challenged with resistant herbivores and problems regarding human health and environmental issues. Indeed, the greatest challenge to modern agriculture is to protect crops from pests and still maintain environmental quality. This study aimed to analyze by in silico, in vitro, and in vivo approaches to the feasibility of using the inhibitory protein extracted from mammals – Bovine Pancreatic Trypsin Inhibitor (BPTI) as a potential inhibitor of digestive trypsins from the pest Anticarsia gemmatalis and comparing the results with the host-plant inhibitor – Soybean Kunitz Trypsin Inhibitor (SKTI). BPTI and SKTI interacts with A. gemmatalis trypsin-like enzyme competitively, through hydrogen and hydrophobic bonds. A. gemmatalis larvae exposed to BPTI did not show two common adaptative mechanisms i.e., proteolytic degradation and overproduction of proteases, presenting highly reduced trypsin-like activity. On the other hand, SKTI-fed larvae did not show reduced trypsin-like activity, presenting overproduction of proteases and SKTI digestion. In addition, the larval survival was reduced by BPTI similarly to SKTI, and additionally caused a decrease in pupal weight. The non-plant protease inhibitor BPTI presents intriguing element to compose biopesticide formulations to help decrease the use of conventional refractory pesticides into integrated pest management programs.