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

Sharma, Pratima; Nath, Amit; Kumari, Reena; Bhardwaj, Satya Vrat

doi:10.1007/s11676-012-0243-7

Cited by 17 publications

(17 citation statements)

References 32 publications

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“…Inhibitors of Kunitz and Bowman-Birk type, isolated from the seeds of Entada acaciifolia and Albizia lebbeck, both belonging to the subfamily Mimosoideae, have molecular weights of 20 and 12.3 kDa, respectively (Oliveira et al 2012, Sharma et al 2012. Based on these results, the inhibitors partially isolated from different species studied include inhibitors of the Kunitz and BowmanBirk type.…”

Section: Discussionmentioning

confidence: 86%

Partial purification of trypsin inhibitors from Parkia seeds (Fabaceae)

Chevreuil

Gonçalves

Calderon³

et al. 2014

Hoehnea

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-(Partial purification of trypsin inhibitors from Parkia seeds (Fabaceae)). Leguminous seeds (Fabaceae) have a high content of inhibitors of which serine protease inhibitors are the most widely studied. However, there are only a few studies related to the investigation of these proteins in tree species belonging to the Amazon flora. The protein content presented in seeds of four Amazonian Leguminosae species, Parkia pendula, P. discolor, P. multijuga and P. Nitida, was extracted by using NaCl 0.15 mol L -1 and then partially fractionated by using affinity chromatography performed on a trypsinSepharose 4B. These inhibitors presented different affinities between trypsin and chymotrypsin serine proteases, showing a higher inhibition to trypsin compared to chymotrypsin, except for P. nitida, which showed high inhibition against both enzymes. The SDS-PAGE analysis showed that the species from Parkia genus have a main band corresponding to partially purified trypsin inhibitors. The apparent molecular mass inhibitors (approximately 13-18 kDa) and the high specificity for trypsin suggest the occurrence of Bowman-Birk and Kunitz type inhibitors. Keywords: Amazonia, Bowman-Birk inhibitor, Kunitz inhibitor, leguminous seeds, serine protease RESUMO -(Purificação parcial de inibidores de tripsina de sementes de Parkia (Fabaceae)). Sementes de leguminosas (Fabaceae) apresentam alto conteúdo de inibidores, incluindo os inibidores de serinoproteinases que têm sido extensivamente estudados. Todavia, poucos estudos foram realizados quanto à investigação dessas proteínas em espécies arbóreas pertencentes à flora amazônica. As proteínas presentes nas sementes de quatro espécies de leguminosas da Amazônia, Parkia pendula, P. discolor, P. multijuga e P. nitida, foram obtidas pela extração usando NaCl 0.15 mol L -1 e, parcialmente purificadas usando a cromatografia de afinidade em tripsina-Sepharose 4B. Os inibidores exibiram afinidades diferentes entre a tripsina e a quimotripsina, exceto para P. nitida, a qual apresentou alta inibição contra as duas enzimas. A análise em SDS-PAGE mostrou que as espécies do gênero Parkia contém uma banda principal correspondendo aos inibidores de tripsina parcialmente purificados. As massas moleculares aparentes determinadas para os inibidores (aproximadamente, 13 a 18 kDa) e a alta especificidade pela tripsina sugerem a ocorrência de inibidores do tipo Bowman-Birk e Kunitz. Palavras-chave: Amazônia, Inibidor Bowman-Birk, Inibidor Kunitz, sementes de leguminosas, serinoproteinase 1.

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Section: Discussionmentioning

confidence: 86%

Partial purification of trypsin inhibitors from Parkia seeds (Fabaceae)

Chevreuil

Gonçalves

Calderon³

et al. 2014

Hoehnea

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“…The noncompetitive mechanism found for the inhibition of ApTI on the trypsin‐like enzymes of A. gemmatalis agrees with other in silico studies carried out with trypsins from other species of the order Lepidoptera (Migliolo et al, 2010; Ramalho et al, 2018), and the reactive (inhibitory) peptide bond was identified in the alpha chain with the participation of the Arg64 residue and amino acid residues around Pro63, Ile65, and Arg66. This bond is at an exactly homologous position to the reactive sites identified in the soybean inhibitor at Arg‐63‐Ile‐64 (McPherson, Daym, & Larson, 2019) in Albizia at Arg‐66‐I1e‐67 (Sharma, Nath, Kumari, & Bhardwaj, 2012) and in Psophocarpus at Arg‐64‐Ser‐65 (Yamamoto, Hara, & Ikenaka, 1983).…”

Section: Discussionmentioning

confidence: 82%

Noncompetitive tight‐binding inhibition ofAnticarsia gemmatalistrypsins byAdenanthera pavoninaprotease inhibitor affects larvae survival

Meriño‐Cabrera

Mendes

Castro

et al. 2020

Arch Insect Biochem Physiol

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The economic loss in soybean crops caused by the Lepidoptera insects has encouraged the search for new strategies to control this pest, which are currently based on synthetic insecticides. This paper evaluated the ability of ApTI (Adenanthera pavonina trypsin inhibitor) to inhibit trypsin-like proteins from Anticarsia gemmatalis by docking, molecular dynamics, and enzymatic and survival assay. The docking and molecular dynamic simulation between trypsin and ApTI were performed using the program CLUSPRO and NAMD, respectively. The inhibitory constant K i and the inhibition type were determined through chromogenic assays. The survival assay of neonatal larvae under treatment with artificial diet supplemented with ApTI was also performed. The ApTI binding site was predicted to block substrate access to trypsin due to four interactions with the enzyme, producing a complex with a surface area of 1,183.7 Å 2 .The kinetic analysis revealed a noncompetitive tightbinding mechanism. The survival curves obtained using Kaplan-Meier estimators indicated that the highest larvae mortality was 60%, using 1.2 mg of ApTI per 100 ml of artificial diet. The in vitro, in vivo, and in silico studies demonstrated that ApTI is a strong noncompetitive inhibitor of trypsin with biotechnological potential for the control of A. gemmatalis insect. K E Y W O R D S binding, insects, Kunitz inhibitor, molecular docking, noncompetitive, trypsin enzyme

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“…On the other hand, proteinase inhibitors are potential candidates for biocontrol of insect pests since insect digestive proteinases are promising targets towards control of various insects (Sharma et al, 2012). Proteases have been found to be effective against many Coleopteran (Elden, 2000).…”

Section: Resultsmentioning

confidence: 99%

Susceptibility response of varieties and local populations of lupines to Bruchus rufimanus (Coleoptera: Chrysomelidae)

Nikolova

2021

AAS

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This study aimed to evaluate the susceptibility response of varieties and local populations of lupines to Bruchus rufimanus in multi-environment field tests. Seed damaged rate and susceptibility index were assessed in each environment and subjected to a heritability-adjusted genotype and genotype x environment biplot analysis. It was found that the susceptibility index of damaged seeds was positively related to precipitation amounts and humidity, and inversely to min and max temperatures. The seed damaged rate was positively related to temperatures but negatively to rain and humidity. The local polish population WAT and cultivars Pink Mutant, Solnechnii, and Bezimenii 1 had the lowest seed damaged rate and stable position across environments. Meanwhile, these cultivars showed a low susceptibility index and low variability. The discrepancy between the early phenological development of ‘Pink Mutant’, ‘Solnechnii’, and ‘Bezimenii 1’ and the life cycle of B. rufimanus was one of the reasons for manifested tolerance. Correlations between damaged seed and susceptibility index as well as the mass of 1000 seeds and sensitivity index were strongly positive and negative, respectively. ‘WAT’, ‘Pink Mutant’, ‘Solnechnii’, and ‘Bezimenii 1’ had a clear advantage in defending itself from B. rufimanus attack, which makes them particularly interesting for breeding purposes.

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

Cited by 17 publications

References 32 publications

Partial purification of trypsin inhibitors from Parkia seeds (Fabaceae)

Partial purification of trypsin inhibitors from Parkia seeds (Fabaceae)

Noncompetitive tight‐binding inhibition ofAnticarsia gemmatalistrypsins byAdenanthera pavoninaprotease inhibitor affects larvae survival

Susceptibility response of varieties and local populations of lupines to Bruchus rufimanus (Coleoptera: Chrysomelidae)

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