Demystifying the Biochemistry: Theoretical Concepts and Practical Activities for High School

INTRODUCTION: The education goes through a crisis in their teaching-learning process, due to the fact that their contents are developed out of context, merely theoretical activities. Several scholars warn about the importance of working with experiential activities to enhance and stimulate student learning, seeking a more meaningful than we have seen in recent times. The objectives of this work are installation of a laboratory for teaching science in public school, ISEPAM, at North of Rio de Janeiro, formation of multipliers agents and promoting a Scientific Week/Exhibition involving all school community. MATERIALS AND METHODS:First, we perform the spatial organization of the laboratory, including purchase of materials/equipment. In a second step, experimental activities involving identification of Macromolecules in foods were prepared and tested. From this, we promote short courses in biochemistry for teachers to forming multiplier agents. As a conclusion of the activities of this project a Scientific Week/Exhibition was promoted in school. RESULTS AND DISCUSSION: Practical classes on fermentation, identification of lipids, proteins and carbohydrates were performed by teachers and students from ISEPAM. We observed in the biochemistry course which the participants have established relationships between theory and practice and also they showed interest in the insertion of experiments in their work routine.. Regarding the Scientific Week/Exhibition, students of elementary and high school participated. They were encouraged and integrated with experiment activities and thereby new talents for scientific and technological interest were identified. CONCLUSION: It was observed that the experimental activities become more attractive and challenging the search for knowledge

Isolation And Characterization Of Isolectins From Talisia Esculenta Seeds

Four isolectins (TEL-I, TEL-II, TEL-III and TEL-IV) were isolated from seeds of Talisia esculenta by reverse-phase high-performance liquid chromatography. RP-HPLC was performed on a u-Bondapack C18 column (0.78 cm X 30 cm) (Waters 991-PDA system) at room temperature. Rechromatography of the four fractions on a C18 column under the same conditions yielded lectins with two dissimilar subunits (M r 20 kDa and 40 kDa) bound noncovalently. The isolectins showed very similar characteristics, such as molecular masses, N-terminal sequences, and hemagglutinating activity, but differed in their isoelectric points and in inhibition by carbohydrates. © 2001 Plenum Publishing Corporation.206495500Abrecht, H., Wattiez, R., Ruysschaert, J.-M., Homblé, F., (2000) Plant Physiol., 124, pp. 1181-1190Animashaun, T., Togun, R.A., Hughes, R.C., (1994) Glycoconjugate J., 11, pp. 299-303Anuradha, P., Bhide, S.V., (1999) Phytochemistry, 52, pp. 751-758Barbieri, L., Falasca, A., Franceschi, C., Licastro, F., Rossi, C.A., Stirpe, F., (1983) Biochem. J., 215, pp. 433-439Blum, H., Beier, H., Gross, H.J., (1987) Electrophoresis, 8, pp. 93-99Cavada, B.S., Santos, C.F., Granjeiros, T.B., Nunes, E.P., Sales, P.V.P., Ramos, R.L., Sousa, F.A.M., Calvete, J.J., (1998) Phytochemistry, 49, pp. 675-680Datta, P.K., Figueroa, M.O.R., Lajolo, F.M., (1993) J. Agric. Food Chem., 41, pp. 1851-1855Dubois, M., Gilles, K.A., Hamilton, J.K., Rebers, P.A., Smith, F., (1956) Anal. Chem., 28, pp. 350-356Eloumani, H., Bladier, D., Caruelle, D., Courty, J., Joubert, R., Caron, M., (1990) Int. J. Biochem., 22, pp. 539-544Freire, M.G.M., Gomes, V.M., Corsini, R.E., Machado, O.L.T., De Simone, S.G., Novello, J.C., Marangoni, S., Macedo, M.L.R., Plant Physiol. Biochem., , in pressHarata, K., Muraki, M., (2000) J. Mol. Biol., 297, pp. 673-681Kilpatrick, D.C., Weston, J., Urbaniak, S.J., (1983) Anal. Biochem., 134, pp. 205-209Laemmli, U.K., (1970) Nature, 227, pp. 680-689Machuka, J.S., Okeola, O.G., Van Damme, J.M., Chrispeels, M.J., Leuven, F.V., Peumans, W.J., (1999) Phytochemistry, 51, pp. 721-728Matsumoto, I., Koyama, T., Kitagaki-Ogawa, H., Seno, N., (1987) J. Chromatog., 400, pp. 77-81Ooi, L.S.M., Ng, T.B., Sun, S.S.M., Ooi, V.E.C., (2000) J. Protein Chem., 19, pp. 163-168Pérez, G., (1998) Int. J. Biochem. Cell Biol., 30, pp. 843-853Pratt, R.C., Singh, N.K., Shade, R.E., Murdock, L.L., Bressan, R.A., (1990) Plant Physiol., 93, pp. 1453-1459Rüdiger, H., (1997) Glycosciences, Status and Perpectives, 2, pp. 415-438. , H-J. Gabius and S. Gabius (Eds): London, Chapman & Hall, WeinheimRüdiger, H., (1998) Acta Anat., 161, pp. 130-152Santos-de-Oliveira, R., Dias-Baruffi, M., Thomaz, S.M.O., Beltramini, L.M., Roque-Barreira, M.C., (1994) J. Immunol., 22, pp. 1798-1807Suzuki, T., Takagi, T., Furukohri, T., Kawamura, K., Nakauchi, M., (1990) J. Biol. Chem., 265, pp. 1274-1281Wongkham, S., Wongkham, C., Boonsiri, P., Simasathiansophon, S., Trisonthi, C., Atisook, K., (1995) Phytochemistry, 40, pp. 1331-133

Purification And Characterization Of A Trypsin Inhibitor From Plathymenia Foliolosa Seeds

A novel trypsin inhibitor (PFTI) was isolated from Plathymenia foliolosa (Benth.) seeds by gel filtration chromatography on a Sephadex G-100, DEAE-Sepharose, and trypsin-Sepharose columns. By SDS-PAGE, PFTI yielded a single band with a Mr of 19 kDa. PFTI inhibited bovine trypsin and bovine chymotrypsin with equilibrium dissociation constants (Ki) of 4 × 10-8 and 1.4 × 10-6 M, respectively. PFTI retained more than 50% of activity at up to 50°C for 30 min, but there were 80 and 100% losses of activity at 60 and 70°C, respectively. DTT affected the activity or stability of PFTI. The N-terminal amino acid sequence of PFTI showed a high degree of homology with various members of the Kunitz family of inhibitors. Anagasta kuehniella is found worldwide; this insect attacks stored grains and products of rice, oat, rye, corn, and wheat. The velvet bean caterpillar (Anticarsia gemmatalis) is considered the main defoliator pest of soybean in Brazil. 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A protein with amino acid sequence homology to bovine insulin is present in the legume Vigna unguiculata (cowpea)

Since the discovery of bovine insulin in plants, much effort has been devoted to the characterization of these proteins and elucidation of their functions. We report here the isolation of a protein with similar molecular mass and same amino acid sequence to bovine insulin from developing fruits of cowpea (Vigna unguiculata) genotype Epace 10. Insulin was measured by ELISA using an anti-human insulin antibody and was detected both in empty pods and seed coats but not in the embryo. The highest concentrations (about 0.5 ng/µg of protein) of the protein were detected in seed coats at 16 and 18 days after pollination, and the values were 1.6 to 4.0 times higher than those found for isolated pods tested on any day. N-terminal amino acid sequencing of insulin was performed on the protein purified by C4-HPLC. The significance of the presence of insulin in these plant tissues is not fully understood but we speculate that it may be involved in the transport of carbohydrate to the fruit

Purification And Characterization Of A New Trypsin Inhibitor From Dimorphandra Mollis Seeds

A second trypsin inhibitor (DMTI-II) was purified from the seed of Dimorphandra mollis (Leguminosae-Mimosoideae) by ammonium sulfate precipitation (30-60%), gel filtration, and ionexchange and affinity chromatography. A molecular weight of 23 kDa was estimated by gel filtration on a Superdex 75 column SDS-PAGE under reduced conditions showed that DMTI-II consisted of a single polypeptide chain, although isoelectric focusing revealed the presence of three isoforms. The dissociation constant of 1.7 ́ 10-9 M with bovine trypsin indicated a high affinity between the inhibitor and this enzyme. The inhibitory activity was stable over a wide pH range and in the presence of DTT. 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Lima bean (Phaseolus lunatus) seed coat phaseolin is detrimental to the cowpea weevil (Callosobruchus maculatus)

The presence of phaseolin (a vicilin-like 7S storage globulin) peptides in the seed coat of the legume Phaseolus lunatus L. (lima bean) was demonstrated by N-terminal amino acid sequencing. Utilizing an artificial seed system assay we showed that phaseolin, isolated from both cotyledon and testa tissues of P. lunatus, is detrimental to the nonhost bruchid Callosobruchus maculatus (F) (cowpea weevil) with ED50 of 1.7 and 3.5%, respectively. The level of phaseolin in the seed coat (16.7%) was found to be sufficient to deter larval development of this bruchid. The expression of a C. maculatus-detrimental protein in the testa of nonhost seeds suggests that the protein may have played a significant role in the evolutionary adaptation of bruchids to legume seeds

Simultaneous allergen inactivation and detoxification of castor bean cake by treatment with calcium compounds

Ricinus communis L. is of great economic importance due to the oil extracted from its seeds. Castor oil has been used for pharmaceutical and industrial applications, as a lubricant or coating agent, as a component of plastic products, as a fungicide or in the synthesis of biodiesel fuels. After oil extraction, a castor cake with a large amount of protein is obtained. However, this by-product cannot be used as animal feed due to the presence of toxic (ricin) and allergenic (2S albumin) proteins. Here, we propose two processes for detoxification and allergen inactivation of the castor cake. In addition, we establish a biological test to detect ricin and validate these detoxification processes. In this test, Vero cells were treated with ricin, and cell death was assessed by cell counting and measurement of lactate dehydrogenase activity. The limit of detection of the Vero cell assay was 10 ng/mL using a concentration of 1.6 x 10(5) cells/well. Solid-state fermentation (SSF) and treatment with calcium compounds were used as cake detoxification processes. For SSF, Aspergillus niger was grown using a castor cake as a substrate, and this cake was analyzed after 24, 48, 72, and 96 h of SSF. Ricin was eliminated after 24 h of SSF treatment. The cake was treated with 4 or 8% Ca(OH)2 or CaO, and both the toxicity and the allergenic properties were entirely abolished. A by-product free of toxicity and allergens was obtained