Red kidney bean seeds (Phaseolus vulgaris) contain a variety of bioactive proteins including lectins, enzyme inhibitors, hydrolytic enzymes and antifungal proteins. The aim of this research was to investigate activities of selected low pH and heat-stable bioactive proteins extracted from different parts of red kidney bean seed, seedling and pod as well as seed and root exudates. Crude red kidney bean seed extracts inhibited growth of Alternaria alternata as well as its protease activity, but not its amylase activity. A protein with inhibitory activity against growth of A. alternata was purified from extracts of the red kidney bean cotyledons and embryonic axis. This purified bean protein was devoid of chitinase and β-1, 3- glucanase activities. Also, it did not inhibit porcine pancreatic α-amylase, bovine trypsin, amylase and protease of A. alternata suggesting that the antifungal activity of the protein is not related to these activities. Proteinaceous extracts of red kidney bean cotyledons induced melanin and conidia formation in mycelium of A. alternata. A protein responsible for this conidiation inducing effect was shown for the first time to be a mannose-binding lectin which is also known as PvFRIL (Phaseolus vulgaris fetal liver tyrosine kinase 3-receptor interacting lectin). An unexpected finding was that extracts of the embryonic axis stimulated rather than inhibited porcine α-amylase activity. Phytohemagglutinin (PHA-L in particular), co-extracted with α-amylase inhibitor from red kidney bean seeds, was implicated as an α-amylase stimulator with the potential of greatly assisting digestion of starch. In cotyledonary extracts, amylase stimulatory activity was masked by amylase inhibitory activity that was inactivated when the extracts were boiled for 10 min. An in-gel non-denaturing electrophoretic method was used to show presence of porcine α-amylase isoinhibitors in extracts of the cotyledons and embryonic axis. All other seedling parts as well as seed and root exudates had amylase stimulatory activity. Another improved non-denaturing electrophoretic method with immobilized azoalbumin was developed for in-gel detection of isoinhibitors of bovine trypsin in seed parts. It eliminates the need for both time-consuming and labourious staining, destaining or renaturation steps used in other methods. Accumulation of most of the selected bioactive proteins during seed development in different seed parts appeared to start at 20 days after flower abscission. The activities of these proteins decreased to lower levels after 11 days of germination. Besides these observed developmental changes, under abiotic (UV-C irradiation) and biotic (seedlings co-cultured with A. alternata) stress, increased activity of some of the selected bioactive proteins were detected. In conclusion, this study has contributed to a better understanding of antifungal activity and the selected bioactive proteins in extracts of red kidney bean.

Made available in DSpace on 2014-12-17T14:03:42Z (GMT). No. of bitstreams: 1 JoycellaneANCR_DISSERT.pdf: 2345415 bytes, checksum: 891d1d3a4e1202d27cf8a25aff1812be (MD5) Previous issue date: 2013-02-19 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / The seeds are excellent sources of proteinase inhibitors and have been highlighted owing to various applications. Among these applications are those in effect on food intake and weight gain that stand out because of the increasing number of obese individuals. This study evaluated the effects of trypsin inhibitor present in the seed of tamarind (Tamarindus indica L.) reduction in weight gain, biochemical and morphological alterations in Wistar rats. For this, we partially purified a trypsin inhibitor tamarind seed. This inhibitor, ITT2 at a concentration of 25 mg / kg body weight, over a period of 14 days was able to reduce food intake in rats (n = 6) by approximately 47%, causing a reduction in weight gain approximately 70% when compared with the control group. With the evaluation of the in vivo digestibility was demonstrated that the animals lost weight due to satiety, presented by the reduction of food intake, since there were significant differences between true digestibility for the control group (90.7%) and the group treated with inhibitor (89.88%). Additionally, we checked the deeds of ITT2 on biochemical parameters (glucose, triglycerides, total cholesterol, high-density lipoprotein, low-density lipoprotein, glutamic-pyruvic transaminase, glutamic oxaloacetic transaminase, gamma glutamyl transferase albumin, globulin, total protein and C-reactive protein) and these, when assessed in the study groups showed no statistically significant variations. We also evaluate the histology of some organs, liver, stomach, intestine, and pancreas, and showed no changes. And to evaluate the effect of trypsin inhibitor on food intake due to the satiety is regulated by cholecystokinin (CCK) were measured plasma levels, and it was observed that the levels of CCK in animals receiving ITT2 were significantly higher ( 20 + 1.22) than in animals receiving only solution with casein (10.14 + 2.9) or water (5.92 + 1.15). Thus, the results indicate that the effect caused ITT2 satiety, reducing food intake, which in turn caused a reduction in weight gain in animals without causing morphological and biochemical changes, this effect caused by the elevation of plasma levels CCK / As sementes s?o excelentes fontes de inibidores de proteinases, e a estes t?m sido atribu?do in?meras aplica??es. Dentre estas aplica??es est?o as com efeito sobre o consumo alimentar e o ganho de peso que se destacam devido o crescente aumento de indiv?duos obesos. Esse estudo avaliou os efeitos do inibidor de tripsina presente na semente de tamarindo (Tamarindus indica L.) sobre o ganho de peso e par?metros bioqu?micos e morfohistol?gicos de ?rg?os de ratos wistar. Para isso, foi parcialmente purificado um inibidor de tripsina de semente de tamarindo (ITT2). Este na concentra??o de 25 mg/Kg de peso, num per?odo de 11 dias, foi capaz de reduzir o consumo alimentar de ratos em cerca de 47%, ocasionando uma redu??o no ganho de peso de aproximadamente 70% quando comparado com o grupo controle. Com a avalia??o da digestibilidade proteica in vivo foi poss?vel demonstrar que os animais perderam peso devido ? saciedade, apresentada pela redu??o do consumo alimentar, visto que n?o houve varia??es significativas da digestibilidade verdadeira entre o grupo controle (90,7%) e o grupo tratado com inibidor (89,88%). Al?m disso, o sangue dos animais foi coletado a fim de serem avaliados os efeitos do ITT2, sobre par?metros bioqu?micos (glicose, triglicer?deos, colesterol total, lipoprote?nas de alta-densidade, lipoprote?na de baixa-densidade, alanina amino transferase, aspartato amino transferase, gama glutamil transferase, albumina, globulina, prote?nas totais e prote?na C reativa) e estes n?o apresentaram varia??es significativas. Os ?rg?os, f?gado, est?mago, intestino, al?m do p?ncreas, n?o apresentaram altera??es. Para avaliar se o efeito do ITT2 sobre o consumo alimentar devido ? saciedade seria acompanhado pelo aumento de colecistocinina (CCK), foram dosados os seus n?veis plasm?ticos, e observado que os n?veis de CCK nos animais que receberam ITT2 eram significativamente maiores (20 + 1,22) do que nos animais que receberam apenas case?na (10,14 + 2,9) ou ?gua (5,92 + 1,15). Dessa forma, os resultados obtidos, indicam que o ITT2 causou um efeito sacietog?nico, reduzindo o consumo alimentar, que por sua vez ocasionou uma redu??o no ganho de peso nos animais, por?m sem causar altera??es bioqu?micas ou morfohistol?gicas, efeito este acompanhado pela eleva??o nos n?veis plasm?ticos de CCK