Partial characterization of proteins from baru (Dipteryx alata Vog) seeds

BACKGROUND: Baru (Dipteryx alata Vog.) is a fruit distributed throughout the Brazilian savanna and contains a seed with a high protein content, whose properties have been rarely explored. The purpose of this study was to characterize this protein, especially by isolation and quantifying its fractions and measuring some of its molecular properties. RESULTS: Baru seeds contain 244 g kg(-1) protein on a dry weight basis. Solubility profiles showed a preponderance of globulins. This fraction dominated the seed composition, with 61.7 wt% of the total soluble proteins. Albumins and glutelins accounted for 14 and 3.3 wt%, respectively. SDS-PAGE resolution of albumin and globulin showed main bands with molecular weights of 84 kDa and 64,66 and 73 kDa, respectively. The total protein of the flour and the globulin showed values of in vitro digestibility of 85.59% and 90.54%, relative to casein. Total globulin produced only one chromatographic peak, both on Sepharose CL-6B gel filtration and on DEAE-cellulose ion-exchange columns, eluted at a concentration of 0.12 mol L(-1) NaCl. CONCLUSION: The baru seed had high protein content with large quantities of storage proteins. The chromatographic and solubility profiles indicate the predominance of a fraction with characteristics of a legumin-type protein. (C) 2011 Society of Chemical Industry

Potential of Amaranthus cruentus BRS Alegria in the production of flour, starch and protein concentrate: chemical, thermal and rheological characterization

BACKGROUND: Amaranth is a little-known culture in Brazilian agriculture. Amaranthus cruentus BRS Alegria was the first cultivar recommended by Embrapa for the soil of the Brazilian scrubland. In order to evaluate the potential of this species in the production of flour, starch and protein concentrates, the latter products were obtained from A. cruentus BRS Alegria seeds, characterized and compared with the products obtained from the A. caudatus species cultivated in its soil of origin. RESULTS: The seeds of A. cruentus BRS Alegria furnished high-purity starch and flour with significant content of starch, proteins, and lipids. The starch and flour of this species presented higher gelatinization temperatures and formed stronger gels upon cooling compared with those obtained from the A. caudatus species. This is due to their greater amylose content and a difference in the composition of the more important fatty acids, such as stearic, oleic and linoleic acids, which indicates that they have greater heat stability. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and differential scanning calorimetry revealed the presence of albumins, globulins, glutelins and prolamins in the protein concentrate, which was obtained as a byproduct of starch production. CONCLUSION: Amaranthus cruentus BRS Alegria has potential application in the production of flour, starch and protein concentrates, with interesting characteristics for use as food ingredients. (C) 2010 Society of Chemical Industry

Chemical and nutritional evaluation of kernels of bocaiuva, Acrocomia aculeata (Jacq.) Lodd.

Protein characterization and results of proximate composition and mineral analyses of fruit kernels of bocaiuva, Acrocomia aculeata (Jacq.) Lodd., are reported. The kernels presented high contents of oil (51.7%), protein (17.6%) and fiber (15.8%). The seeds´ soluble proteins were isolated according to their solubility. The main separated proteins were globulins (53.5%) and glutelins (40.0%). Moreover, the presence of low molecular mass proteases in these two fractions was shown by the SDS-PAGE method. The assays of protease-inhibitory and hemagglutinating activities showed that bocaiuva´s protein fractions were not resistant to trypsin or chymotrypsin activities and that both had low lectin content. The globulin in vitro digestibility assay resembled a casein standard. Neither globulin nor glutelin enzymatic hydrolyses increased significantly (p < 0.05) after heat treatment. Threonine and lysine are the most limiting amino acids, respectively from two major protein fractions of the bocaiuva kernel, globulin (47.1% amino acid score) and glutelin (49.5% amino acid score), in terms of the theoretical profiles for children in the age range of 2 to 5 years recommended by the FAO/WHO. Bocaiuva kernels are found to be rich in calcium, phosphorus and manganese compared to some fruit nuts such as cashew and coconut.

Isolation, fractionation and characterisation of proteins from Mucuna bean

The chemical composition and fractional distribution of protein isolates prepared from species of Mucuna bean were studied. Using six different extraction media, the yield of protein based on the Kjeldahl procedure varied from 8% to 34%, and the protein content varied from 75% to 95%. When the yields were high, the colour of the isolates generally tended to be dark and unsatisfactory. Hence, the use of chemical treatments and high pressure processing were explored. The solubility maxima for the protein isolates in water were found to occur at pH values of 2.0 and 11.0, while the pH corresponding to minimum solubility (i.e. isoelectric region) occurred at pH values of 4.0 and 5.0. The total essential amino acid in the isolates ranged from 495 to 557 mg g(-1) protein, which compares favourably with the recommended level for pre-school and school children. Methionine and cysteine were the limiting amino acids. A key nutritional attribute of the protein isolates was its high lysine content. The isolate can therefore complement cereal-based foods which are deficient in lysine. The proteins mainly consisted of albumins, glutelins and globulins. Prolamins were only present in trace concentration (< 0.3%). Gel filtration chromatograms of the isolates indicated the presence of major protein fractions with molecular weights of 40 and 15 kDa, while gel electrophoresis (SDS-PAGE) indicated a major broad zone with molecular weights of 36 +/- 7 and 17.3 +/- 13 kDa. (c) 2006 Elsevier Ltd. All rights reserved.

The trafficking pathway of a wheat storage protein in transgenic rice endosperm

Background and Aims The trafficking of proteins in the endoplasmic reticulum (ER) of plant cells is a topic of considerable interest since this organelle serves as an entry point for proteins destined for other organelles, as well as for the ER itself. In the current work, transgenic rice was used to study the pattern and pathway of deposition of the wheat high molecular weight (HMW) glutenin sub-unit (GS) 1Dx5 within the rice endosperm using specific antibodies to determine whether it is deposited in the same or different protein bodies from the rice storage proteins, and whether it is located in the same or separate phases within these. Methods The protein distribution and the expression pattern of HMW sub-unit 1Dx5 in transgenic rice endosperm at different stages of development were determined using light and electron microscopy after labelling with antibodies. Key results The use of HMW-GS-specific antibodies showed that sub-unit 1Dx5 was expressed mainly in the sub-aleurone cells of the endosperm and that it was deposited in both types of protein body present in the rice endosperm: derived from the ER and containing prolamins, and derived from the vacuole and containing glutelins. In addition, new types of protein bodies were also formed within the endosperm cells. Conclusions The results suggest that the HMW 1Dx5 protein could be trafficked by either the ER or vacuolar pathway, possibly depending on the stage of development, and that its accumulation in the rice endosperm could compromise the structural integrity of protein bodies and their segregation into two distinct populations in the mature endosperm.

Efeito do consumo de farinha de linhaça (Linum usitatissimum) no crescimento de ratos Wistar e relação com a digestibilidade de globulinas e fatores antinutricionais protéicos

Linseed is an important oilseed consumed raw as nutritional supplement, that although represents a rich source of nutrients, its nutritional value could be impaired due to the presence of antinutritional factors. In this study, protein fractions from raw linseed flour were extracted and isolated being obtained 12% of albumins, 82% of globulins, 5% of glutelins and 1% of prolamins. These proteins were visualized by SDS-PAGE and albumins showed low molecular mass protein bands around 21 kDa and minor bands, similar to that of trypsin inhibitor; Globulins presented protein bands with high molecular masses, which possibly are constituents of multimeric proteins, such as legumins. After determination of the centesimal composition of raw linseed, it was used as exclusive protein source for young rats to evaluate its effect on animal growth. The results showed negative effects on rat growth (weight gain 73% less than the control group) and reduction of intestinal villus (35%), that could be related with in vitro and in vivo globulin digestibility and proteinaceous antinutritional factors (mammalian digestive enzymes inhibitors and lectins) in albumin fraction. Native globulins showed, by SDS-PAGE, low susceptibility in vitro to trypsin and chymotrypsin, however presented high degradation by pancreatin. Thermal treatment of globulins for 5 and 15 minutes at 100ºC improved considerably its digestibility by trypsin and pancreatin. Globulins presented 93.2% in vivo digestibility, similar to the control protein. Albumin fraction had high trypsin inhibition activity (100%) and chymotrypsin inhibition of 28.3%; haemagglutinating activity was not detected. The results of this study indicate the negative action of trypsin inhibitors on animal growth, but can not be discarded its combined action with other antinutritional factors, which could compromise the raw linseed utilization as an alternative food

DSC studies on protein isolate of guava seeds Psidium guajava

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Partial characterization of proteins from baru (Dipteryx alata Vog) seeds

BACKGROUND: Baru (Dipteryx alata Vog.) is a fruit distributed throughout the Brazilian savanna and contains a seed with a high protein content, whose properties have been rarely explored. The purpose of this study was to characterize this protein, especially by isolation and quantifying its fractions and measuring some of its molecular properties.RESULTS: Baru seeds contain 244 g kg(-1) protein on a dry weight basis. Solubility profiles showed a preponderance of globulins. This fraction dominated the seed composition, with 61.7 wt% of the total soluble proteins. Albumins and glutelins accounted for 14 and 3.3 wt%, respectively. SDS-PAGE resolution of albumin and globulin showed main bands with molecular weights of 84 kDa and 64,66 and 73 kDa, respectively. The total protein of the flour and the globulin showed values of in vitro digestibility of 85.59% and 90.54%, relative to casein. Total globulin produced only one chromatographic peak, both on Sepharose CL-6B gel filtration and on DEAE-cellulose ion-exchange columns, eluted at a concentration of 0.12 mol L(-1) NaCl.CONCLUSION: The baru seed had high protein content with large quantities of storage proteins. The chromatographic and solubility profiles indicate the predominance of a fraction with characteristics of a legumin-type protein. (C) 2011 Society of Chemical Industry

Thermal study and physico-chemical characterization of some functional properties of guava seeds protein isolate (Psidium guajava)

The guava seed protein isolate ( PI) was obtained from the protein precipitation belonging to the class of the gluteline (Ip 4.5). The conditions for the preparation of the PI were determined by both the solubility curve and simultaneous thermogravimetry-differential thermal analysis (TG-DTA): pH 11.5, absence of NaCl and whiteners and T=( 25 +/- 3) degrees C. Under these conditions a yield of 77.0 +/- 0.4%, protein content of 94.2 +/- 0.3, ashes 0.50 +/- 0.05% and thermal stability, T= 200 degrees C, were obtained. The TG-DTA curves and the PI emulsification capacity study showed the presence of hydrophobic microdomains at pH 11.5 and 3.0 suggesting a random coil protein conformation and, to pH 10.0, an open protein conformation. The capacity of emulsification (CE), in the absence of NaCl, was verified for: 1 - pH 3.0 and 8.5, using the IP extracted at pH 10.0 and 11.5, CE >= 343 +/- 5 g of emulsified oil/g of protein; 2 - pH 6.60 just for the PI obtained at pH 11.5, CE >= 140 +/- 8 g of emulsified oil/g of protein.

Proteína e qualidade de sementes de feijão (Phaseolus vulgaris L.) em função da adubação nitrogenada em plantio direto

Nitrogen limitation in the common bean (Phaseolus vulgaris L.) produces reduced productivity and seeds with low protein content and physiological quality. We assessed the effects of nitrogen fertilizer side dressing on the protein content and physiological quality of the seeds of P. vulgaris L. cultivars (cv.) IPR Juriti and Pérola grown in plots (soil type = Dystrophic Red Latosol) using a no-tillage system under a thick mulch of millet residues at the Experimental Station of UNESP-Ilha Solteira campus, located in Selvíria, MS (Köppen climate type = Aw) during autumn (March/June) and winter (June/September) 2005. For each cultivar, a randomized block experimental design was used with four replicates and factorially arranged treatments equivalent to 0, 30, 60, 90 and 120 kg.ha-1 added nitrogen (as urea, containing 45% of N) applied as a side dressing during the V4-3 and V4-6 phenological stage corresponding to the 3rd and 6th completely opened trifoliolate leaf on the main stem. Supplementation with up to 120 kg.ha-1 nitrogen promoted a greater increase in crude protein at V4-3 (unsupplemented = 17.6% and 16.3%; 120 kg.ha-1 N = 24.1% and 22.3% for cv. IPR Juriti and Pérola, respectively) than at V4-6 (unsupplemented = 19.2% and 18.3%; 120 kg.ha-1 N = 21.3% and 20.3% for cv. IPR Juriti and Pérola, respectively). About 90% of the crude protein from cv. IPR Juriti was composed of soluble protein compared to 72% for the cv. Pérola. Albumins and globulins represented about 80% of the soluble protein and prolamins were lower at 0.6%. In conclusion, nitrogen fertilization up to 120 kg.ha-1 applied as a side dressing at the V4-3 phenological stage in no-tillage under a thick mulch of millet promoted a greater increase of crude protein in common bean seeds than at the V4-6 stage. The highest accumulation of soluble protein occurred at 90 kg.ha-1 applied nitrogen without having a significant influence on the physiological quality of the seeds.

Isolado protéico de farinha de semente de goiaba (Psidium guajava): caracterização de propriedades funcionais e térmicas

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Cooking Effects on Iron and Proteins Content of Beans (Phaseolus Vulgaris L.) by GF AAS and MALDI-TOF MS

The effects of domestic cooking on proteins, organic compounds and Fe distribution in beans (Phaseolus vulgaris L.) were investigated. Sequential extraction with different extractant solutions (mixture of methanol and chloroform 1:2 v/v, water, 0.5 mol L-1 NaCl, 70% v/v ethanol and 0.5 mol L-1 NaOH) were used for extracting lipids, albumins, globulins, prolamins and glutelins, respectively. Iron determination by graphite furnace atomic absorption spectrometry (GF AAS), proteins by Bradford method and organic compounds by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) were carried out in this work. High concentration of albumins, globulins and glutelins were found in raw beans, while in the cooked beans, albumins and glutelins are main proteins types. The MALDI-TOF MS spectra of raw and cooked beans revealed that the domestic cooking altered the molecular weight of the organic compounds, since that in the cooked beans were found compounds between 2 and 3.5 kDa, which were not presented in the raw beans. Besides this, in cooked beans were also observed the presence of four compounds of high molecular weight (12-16 kDa), being that in the raw grains there is only one (ca. 15.2 kDa). In raw grains is possible to observe that Fe is mainly associated to albumins, globulins and glutelins. For cooked grains, Fe is associated to albumins and globulins.

Cooking effects on iron and proteins content of beans (Phaseolus Vulgaris L.) by GF AAS and MALDI-TOF MS

The effects of domestic cooking on proteins, organic compounds and Fe distribution in beans (Phaseolus vulgaris L.) were investigated. Sequential extraction with different extractant solutions (mixture of methanol and chloroform 1:2 v/v, water, 0.5 mol L-1 NaCl, 70% v/v ethanol and 0.5 mol L-1 NaOH) were used for extracting lipids, albumins, globulins, prolamins and glutelins, respectively. Iron determination by graphite furnace atomic absorption spectrometry (GF AAS), proteins by Bradford method and organic compounds by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) were carried out in this work. High concentration of albumins, globulins and glutelins were found in raw beans, while in the cooked beans, albumins and glutelins are main proteins types. The MALDI-TOF MS spectra of raw and cooked beans revealed that the domestic cooking altered the molecular weight of the organic compounds, since that in the cooked beans were found compounds between 2 and 3.5 kDa, which were not presented in the raw beans. Besides this, in cooked beans were also observed the presence of four compounds of high molecular weight (12-16 kDa), being that in the raw grains there is only one (ca. 15.2 kDa). In raw grains is possible to observe that Fe is mainly associated to albumins, globulins and glutelins. For cooked grains, Fe is associated to albumins and globulins.

Efeito do consumo de farinha de linhaça (Linum usitatissimum) no crescimento de ratos Wistar e relação com a digestibilidade de globulinas e fatores antinutricionais protéicos

Linseed is an important oilseed consumed raw as nutritional supplement, that although represents a rich source of nutrients, its nutritional value could be impaired due to the presence of antinutritional factors. In this study, protein fractions from raw linseed flour were extracted and isolated being obtained 12% of albumins, 82% of globulins, 5% of glutelins and 1% of prolamins. These proteins were visualized by SDS-PAGE and albumins showed low molecular mass protein bands around 21 kDa and minor bands, similar to that of trypsin inhibitor; Globulins presented protein bands with high molecular masses, which possibly are constituents of multimeric proteins, such as legumins. After determination of the centesimal composition of raw linseed, it was used as exclusive protein source for young rats to evaluate its effect on animal growth. The results showed negative effects on rat growth (weight gain 73% less than the control group) and reduction of intestinal villus (35%), that could be related with in vitro and in vivo globulin digestibility and proteinaceous antinutritional factors (mammalian digestive enzymes inhibitors and lectins) in albumin fraction. Native globulins showed, by SDS-PAGE, low susceptibility in vitro to trypsin and chymotrypsin, however presented high degradation by pancreatin. Thermal treatment of globulins for 5 and 15 minutes at 100ºC improved considerably its digestibility by trypsin and pancreatin. Globulins presented 93.2% in vivo digestibility, similar to the control protein. Albumin fraction had high trypsin inhibition activity (100%) and chymotrypsin inhibition of 28.3%; haemagglutinating activity was not detected. The results of this study indicate the negative action of trypsin inhibitors on animal growth, but can not be discarded its combined action with other antinutritional factors, which could compromise the raw linseed utilization as an alternative food

Efeito do consumo de farinha de linhaça (Linum usitatissimum) no crescimento de ratos Wistar e relação com a digestibilidade de globulinas e fatores antinutricionais protéicos

Linseed is an important oilseed consumed raw as nutritional supplement, that although represents a rich source of nutrients, its nutritional value could be impaired due to the presence of antinutritional factors. In this study, protein fractions from raw linseed flour were extracted and isolated being obtained 12% of albumins, 82% of globulins, 5% of glutelins and 1% of prolamins. These proteins were visualized by SDS-PAGE and albumins showed low molecular mass protein bands around 21 kDa and minor bands, similar to that of trypsin inhibitor; Globulins presented protein bands with high molecular masses, which possibly are constituents of multimeric proteins, such as legumins. After determination of the centesimal composition of raw linseed, it was used as exclusive protein source for young rats to evaluate its effect on animal growth. The results showed negative effects on rat growth (weight gain 73% less than the control group) and reduction of intestinal villus (35%), that could be related with in vitro and in vivo globulin digestibility and proteinaceous antinutritional factors (mammalian digestive enzymes inhibitors and lectins) in albumin fraction. Native globulins showed, by SDS-PAGE, low susceptibility in vitro to trypsin and chymotrypsin, however presented high degradation by pancreatin. Thermal treatment of globulins for 5 and 15 minutes at 100ºC improved considerably its digestibility by trypsin and pancreatin. Globulins presented 93.2% in vivo digestibility, similar to the control protein. Albumin fraction had high trypsin inhibition activity (100%) and chymotrypsin inhibition of 28.3%; haemagglutinating activity was not detected. The results of this study indicate the negative action of trypsin inhibitors on animal growth, but can not be discarded its combined action with other antinutritional factors, which could compromise the raw linseed utilization as an alternative food