The Experts below are selected from a list of 186 Experts worldwide ranked by ideXlab platform
Hiroyasu Inoue - One of the best experts on this subject based on the ideXlab platform.
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secretion of miraculin through the function of a signal peptide conserved in the kunitz type Soybean Trypsin Inhibitor family
FEBS Letters, 2013Co-Authors: Ayako Takai, Makiko Satoh, Tomomi Matsuyama, Rieko Nakata, Takashi Aoyama, Hiroyasu InoueAbstract:Miraculin, a glycoprotein that modifies sour tastes into sweet ones, belongs to the Kunitz-type Soybean Trypsin Inhibitor (STI) family. To clarify the functional relation of miraculin with Kunitz-type STIs, we investigated its subcellular localization and Trypsin Inhibitory activity. In transgenic Arabidopsis thaliana, miraculin, fused to yellow fluorescent protein, localized to and outside the plasma membrane depending on the putative secretion signal peptide. When transgenic seedlings were cultured in liquid medium, miraculin was present in the supernatant only after cellulase treatment. No Trypsin Inhibitory activity was detected in native or recombinant miraculin. In conclusion, miraculin is secreted outside the plasma membrane through the function of a signal peptide, conserved in Kunitz-type STIs, whereas its Trypsin Inhibitory activity may be lost during its evolution.
H C Sharma - One of the best experts on this subject based on the ideXlab platform.
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biological activity of Soybean Trypsin Inhibitor and plant lectins against cotton bollworm legume pod borer helicoverpa armigera
Plant Biotechnology, 2005Co-Authors: Sonali Shukla, Richa Arora, H C SharmaAbstract:The noctuid, Helicoverpa armigera, is the most important crop pest worldwide. We evaluated the biological activity of Soybean Trypsin Inhibitor and plant lectins against this pest to identify toxin genes for deployment through transgenic plants. Of the seven plant lectins tested, chickpea and snowdrop lectins showed marked antibiotic effects in terms of insect survival and development. Larval survival was lower in artificial diet impregnated with Soybean Trypsin Inhibitor (49%), and snowdrop (64%) and chickpea (65%) lectins compared to untreated control diet (90%). Pupal weight was adversely affected by chickpea lectin (272.6 mg) compared as to untreated control (335.4 mg). Lower pupation and/or adult emernence (� 50%) was observed in diets impregnated with Soybean Trypsin Inhibitor and chickpea, snowdrop and peanut lectins as compared to 90% pupation/adult emergence on untreated control diet. Soybean Trypsin Inhibitor, and lectins from snowdrop, peanut, and chickpea can be considered for deployment through transgenic plants for the management of H. armigera.
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Biological activity of Soybean Trypsin Inhibitor and plant lectins against cotton bollworm/legume pod borer, Helicoverpa armigera
Plant Biotechnology, 2005Co-Authors: Sonali Shukla, Richa Arora, H C SharmaAbstract:The noctuid, Helicoverpa armigera, is the most important crop pest worldwide. We evaluated the biological activity of Soybean Trypsin Inhibitor and plant lectins against this pest to identify toxin genes for deployment through transgenic plants. Of the seven plant lectins tested, chickpea and snowdrop lectins showed marked antibiotic effects in terms of insect survival and development. Larval survival was lower in artificial diet impregnated with Soybean Trypsin Inhibitor (49%), and snowdrop (64%) and chickpea (65%) lectins compared to untreated control diet (90%). Pupal weight was adversely affected by chickpea lectin (272.6 mg) compared as to untreated control (335.4 mg). Lower pupation and/or adult emernence (� 50%) was observed in diets impregnated with Soybean Trypsin Inhibitor and chickpea, snowdrop and peanut lectins as compared to 90% pupation/adult emergence on untreated control diet. Soybean Trypsin Inhibitor, and lectins from snowdrop, peanut, and chickpea can be considered for deployment through transgenic plants for the management of H. armigera.
Fernando L Garciacarreno - One of the best experts on this subject based on the ideXlab platform.
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the protease based compensatory mechanism to minimize the effect of dietary Soybean Trypsin Inhibitor in litopenaeus vannamei
Aquaculture, 2019Co-Authors: Liliana Rojoarreola, Julio H Cordovamurueta, Cyril Choquet, Fernando L GarciacarrenoAbstract:Abstract Soybean meal is a recurrent ingredient in shrimp feed formulations; although economically convenient, the downside is the antinutritional molecules often present in its active forms. Recently, a midgut gland proteolytic activity adjustment to compensate the effect of increasing concentrations of Soybean Trypsin Inhibitor (SBTI) delivered in shrimp feed was found in Litopenaeus vannamei; which prompted us to inquire on the transcriptional response of shrimp digestive peptidases against exogenous peptidase Inhibitors and connect the results for a broader comprehension of the compensatory mechanism triggered by ingestion of SBTI. Our results indicate that such response involves at least two stages: in the first hours, a drastic increase in transcription of Trypsin, an Inhibitor sensitive protease was observed followed by a gradual increase in transcription of putatively less-sensitive protease chymoTrypsin, and a non-sensitive metalloprotease was observed. At 23 h a second doses of SBTI was administered and at 24 h, Trypsin transcripts returned to basal levels, whereas the transcription of genes encoding Inhibitor-insensitive peptidases increased. Taken together, this paper is the first comprehensive study of the compensatory mechanisms to exogenous protease Inhibitors in Penaeidae. Specific activity and gene expression of digestive proteases observe the same pattern, indicating a phenotypic plasticity in the digestive system of L. vannamei as an adaptive response to compensate the protein digestion capacity when some of the peptidase activities are reduced by the presence of protease Inhibitors.
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peptidase compensation in the digestive system of whiteleg shrimp penaeus vannamei against dietary kunitz type Soybean Trypsin Inhibitor
Aquaculture Nutrition, 2017Co-Authors: Claudia I Maytorenaverdugo, Julio H Cordovamurueta, Fernando L GarciacarrenoAbstract:The aim of this study was to assess the regulatory process of digestive peptidases of crustaceans in the presence of Soybean Trypsin Inhibitor (SBTI). This naturally occurring Inhibitor in Soybean meal was used to inhibit the activity of digestive serine peptidases of the whiteleg shrimp Litopenaeus vannamei. In vitro, SBTI inhibited the total proteolytic activity and chymoTrypsin activity by 65%. Trypsin activity was reduced by SBTI from 40% to 15% from 2 to 4 hr of incubation, which is the average time of residence of feed in the shrimp digestive system. During the bioassays, experimental groups were fed with increasing concentration of supplemental SBTI (1 g kg−1 and 2 g kg−1) and digestive gland and faeces of individual specimens were collected daily. At the end of the bioassay, peptidase activity of digestive gland and faeces was shown, revealing differential inhibition after feeding for 5 days. Several serine peptidases were observed in zymograms, showing a compensation effect on the digestive gland through the activation of peptidases from different catalytic type. These results provide evidence that the shrimp digestive gland can overcome the effect of SBTI by two adaptive mechanisms: synthesis of additional peptidases of the serine class and other unidentified peptidases.
Ayako Takai - One of the best experts on this subject based on the ideXlab platform.
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secretion of miraculin through the function of a signal peptide conserved in the kunitz type Soybean Trypsin Inhibitor family
FEBS Letters, 2013Co-Authors: Ayako Takai, Makiko Satoh, Tomomi Matsuyama, Rieko Nakata, Takashi Aoyama, Hiroyasu InoueAbstract:Miraculin, a glycoprotein that modifies sour tastes into sweet ones, belongs to the Kunitz-type Soybean Trypsin Inhibitor (STI) family. To clarify the functional relation of miraculin with Kunitz-type STIs, we investigated its subcellular localization and Trypsin Inhibitory activity. In transgenic Arabidopsis thaliana, miraculin, fused to yellow fluorescent protein, localized to and outside the plasma membrane depending on the putative secretion signal peptide. When transgenic seedlings were cultured in liquid medium, miraculin was present in the supernatant only after cellulase treatment. No Trypsin Inhibitory activity was detected in native or recombinant miraculin. In conclusion, miraculin is secreted outside the plasma membrane through the function of a signal peptide, conserved in Kunitz-type STIs, whereas its Trypsin Inhibitory activity may be lost during its evolution.
Munishwar N. Gupta - One of the best experts on this subject based on the ideXlab platform.
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immobilized metal affinity chromatography without chelating ligands purification of Soybean Trypsin Inhibitor on zinc alginate beads
Biotechnology Progress, 2002Co-Authors: Munishwar N. Gupta, Sulakshana JainAbstract:Immobilized metal affinity chromatography (IMAC) is a widely used technique for bioseparation of proteins in general and recombinant proteins with polyhistidine fusion tags in particular. An expensive and critical step in this process is coupling of a chelating ligand to the chromatographic matrix. This chelating ligand coordinates metal ions such as Cu 2+ , Zn 2+ , and Ni 2+ , which in turn bind proteins. The toxicity of chemicals required for coupling and their slow release during the separation process are of considerable concern. This is an important issue in the context of purification of proteins/enzymes which are used in food processing or pharmaceutical purposes. In this work, a simpler IMAC design is described which should lead to a paradigm shift in the application of IMAC in separation. It is shown that zinc alginate beads (formed by chelating alginate with Zn 2+ directly) can be used for IMAC. As proof of concept, Soybean Trypsin Inhibitor was purified 18-fold from its crude extract with 90% recovery of biological activity. The dynamic binding capacity of the packed bed was 3919 U mL -1 , as determined by frontal analysis. The media could be regenerated with 8 M urea and reused five times without any appreciable loss in its binding capacity.
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imidazole a new ligand for metal affinity precipitation precipitation of kunitz Soybean Trypsin Inhibitor using cu ii loaded copolymers of 1 vinylimidazole with n vinylcaprolactam or n isopropylacrylamide
Applied Biochemistry and Biotechnology, 1997Co-Authors: Yu I Galaev, Munishwar N. Gupta, Anil Kumar, Ritu Agarwal, Bo MattiassonAbstract:Kunitz Soybean Trypsin Inhibitor (STI) was specifically coprecipitated during precipitation of Cu(II)-loaded copolymers induced by increase in temperature and ionic strength. The copolymers used consisted of 1-vinylimidazole andN-vinylcaprolactam orN- isopropylacrylamide. The elution of STI was achieved by solubilization of the STI-Cu(II)-polymer complex in the presence of an excess of the competing ligand, imidazole, and a subsequent precipitation of the polymer with STI remaining free in solution in a purified form as judged by Sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE). To the best of our knowledge this is the first reported successful metal affinity precipitation of protein in a heterobifunctional format.
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Affinity precipitation of Trypsin with Soybean Trypsin Inhibitor linked Eudragit S-100
Journal of Biotechnology, 1994Co-Authors: Ashwani Kumar, Munishwar N. GuptaAbstract:Abstract Soybean Trypsin Inhibitor linked to Eudragit S-100 was used for the affinity precipitation of Trypsin. Polymer and ligand concentrations used in conjugate preparation showed remarkable effect on the Trypsin recovery. Trypsin precipitation efficiency amounted to 89% and recovery was 74%. The final purification of relatively crude commercial Trypsin resulted in 1.85-fold purification. The SDS-PAGE analysis indicated significant purification. The precipitated enzyme activity was around 96% and recovered enzyme activity was 83%.