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Papain is a cysteine endopeptidase of vegetal origin (papaya (Carica papaya L.) with diverse applications in food technology. In this review we have focused our attention on its application in the production of bio-peptides by hydrolysis of proteins from fish residues. This way, a residual material, that can become a contaminant if dumped without control, is converted into highly interesting products. The main bioactivity of the produced peptides is their antioxidant activity, followed by their nutritional and functional activities, but peptides with many other bioactivities have been produced. Thera are also examples of production of hydrolysates with several bioactivities. The enzyme may be used alone, or in combination with other enzymes to increase the degree of hydrolysis. We gratefully recognize the support from the Ministerio de Ciencia e Innovación from Spanish Government (project number CTQ2017-86170-R) and CSIC for the project AEP045. The FPU fellowship (Ministerio de Educacion) for Mr. Morellon–Sterling is gratefully recognized. Dr. Tacias-Pascacio thanks the financial support from “Programa para el Desarrollo Profesional Docente” (PRODEP) from Mexican Government. ABM would like to thank Ministerio de Ciencia Innovación y Universidades and FEDER (Project RTI2018-095291-B-I00) and the Generalitat Valenciana (PROMETEOII/2018/076) for financial support.

Papain is a cysteine protease from papaya, with many applications due to its broad specificity. This paper reviews for first time the immobilization of papain on different supports (organic, inorganic or hybrid supports) presenting some of the features of the utilized immobilization strategies (e.g., epoxide, glutaraldehyde, genipin, glyoxyl for covalent immobilization). Special focus is placed on the preparation of magnetic biocatalysts, which will permit the simple recovery of the biocatalyst even if the medium is a suspension. Problems specific to the immobilization of proteases (e.g., steric problems when hydrolyzing large proteins) are also defined. The benefits of a proper immobilization (enzyme stabilization, widening of the operation window) are discussed, together with some artifacts that may suggest an enzyme stabilization that may be unrelated to enzyme rigidification. We gratefully recognize the support from the Ministerio de Ciencia e Innovación from Spanish Government (project number CTQ2017-86170-R) and CSIC for the project AEP045. The FPU fellowship (Ministerio de Educacion) for Mr. Morellon–Sterling is gratefully recognized. Dr. Tacias-Pascacio thanks the financial support from “Programa para el Desarrollo Profesional Docente” (PRODEP) from Mexican Government. ABM would like to thank Ministerio de Ciencia Innovación y Universidades and FEDER (Project RTI2018-095291-B-I00) and the Generalitat Valenciana (PROMETEOII/2018/076) for financial support.

Silica has been extracted from rice husks via a simple hydrothermal process and functionalized with triethoxy(octyl)silane −OCTES (Octyl–SiO2) and (3-aminopropyl)triethoxysilane – 3-APTES (Amino–SiO2), with the aim of using it as support to immobilize lipase from Thermomyces lanuginosus (TLL) via adsorption. The supports have been characterized by particle size distribution and elemental analyses, XRD, TGA, SEM, AFM and N2 physisorption so as to confirm their functionalization. Effect of pH, temperature, initial protein loading and contact time on the adsorption process has been systematically evaluated. Maximum immobilized protein loading of 12.3 ± 0.1 mg/g for Amino–SiO2 (5 mM buffer sodium acetate at pH 4.0, 25 °C and initial protein loading of 20 mg/g) and 21.9 ± 0.1 mg/g for Octyl–SiO2 (5 mM buffer sodium acetate at pH 5.0, 25 °C and initial protein loading of 30 mg/g) was observed. However, these biocatalysts presented similar catalytic activity in olive oil emulsion hydrolysis (between 630 and 645 U/g). TLL adsorption was a spontaneous process involving physisorption. Experimental data on Octyl–SiO2 and Amino–SiO2 adsorption were well-fitted to the Langmuir isotherm model. It was also investigated whether these biocatalysts could synthesize cetyl esters via esterification reaction. Thus, it was found that cetyl stearate synthesis required 100–110 min of reaction time to attain maximum conversion percentage (around 94%). Ester productivity of immobilized TLL on Amino–SiO2 was 1.3–3.1 times higher than Octyl–SiO2.

This review aims to cover the uses of the commercially available protease Alcalase in the production of biologically active peptides since 2010. Immobilization of Alcalase has also been reviewed, as immobilization of the enzyme may improve the final reaction design enabling the use of more drastic conditions and the reuse of the biocatalyst. That way, this review presents the production, via Alcalase hydrolysis of different proteins, of peptides with antioxidant, angiotensin I–converting enzyme inhibitory, metal binding, antidiabetic, anti-inflammatory and antimicrobial activities (among other bioactivities) and peptides that improve the functional, sensory and nutritional properties of foods. Alcalase has proved to be among the most efficient proteases for this goal, using different protein sources, being especially interesting the use of the protein residues from food industry as feedstock, as this also solves nature pollution problems. Very interestingly, the bioactivities of the protein hydrolysates further improved when Alcalase is used in a combined way with other proteases both in a sequential way or in a simultaneous hydrolysis (something that could be related to the concept of combi-enzymes), as the combination of proteases with different selectivities and specificities enable the production of a larger amount of peptides and of a smaller size. We gratefully recognize the support from the Ministerio de Ciencia e Innovación from Spanish Government (project number CTQ2017-86170-R). The FPU fellowship (Ministerio de Educacion) for Mr. Morellon–Sterling and the fellowship for Mr. Siar from the Algerian Ministry of Higher Education and Scientific Research are also thanked. Dr. Tacias-Pascacio thanks the financial support from “Programa para el Desarrollo Profesional Docente” (PRODEP) from Mexican Government.

Chocolate is an important source of free bioactive amines and amino acids which play important roles in human health. Considering the limited information on the bioaccessibility of these compounds from chocolate, the objective of this study was to characterize their profiles and bioaccessibility in 70% cocoa dark chocolate through in vitro simulation of oral, gastric and intestinal digestions. Seven amines were detected; polyamines were predominant before in vitro digestion, whereas tyramine, cadaverine and spermidine after digestion. All amines showed high bioaccessibility with slight influence of digestive enzymes. Amines increased after gastrointestinal digestion: tyramine (13-fold), tryptamine (9-fold), others (2.4–4.2-fold) and histamine appeared. All amino acids, GABA and ammonia were detected in chocolate, and their contents increased after in vitro digestion due to digestive enzymes (4.6, 2.8 and 2.1, respectively). Dark chocolate protein is a good source of tryptophan, phenylalanine + tyrosine, isoleucine, histidine, but limiting for lysine, leucine, and threonine.

CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico FAPEMIG - Fundação de Amparo à Pesquisa do Estado de Minas Gerais CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior Mushrooms are significant sources of amino acids and bioactive amines; however, their bioaccessibility can be affected by processing and during in vitro digestion. Fresh Agaricus bisporus mushroom was submitted to cooking and canning and samples were submitted to in vitro gastric and gastric-intestinal digestions. An UHPLC method was used for the simultaneous determination of 18 free amino acids, 10 biogenic amines and ammonia in the samples. Fresh mushroom contained 14 free amino acids, with predominance of alanine and glutamic acid; spermidine was the only amine detected; and ammonia was also detected. Spermidine levels were not affected by cooking, but there was a significant loss (14%) after canning. Spermidine levels were not affected by the in vitro gastric and intestinal digestion, suggesting full bioaccessibility. There was a significant decrease on total amino acids levels after cooking and canning, with higher losses for aspartic and glutamic acids in cooked and for aspartic acid and valine in canned mushrooms. After the in vitro gastric and intestinal digestions, the total levels of amino acids increased and two additional amino acids (arginine and methionine) were detected. During in vitro digestion many of the amino acids were released mainly in the intestinal phase. After in vitro digestion, amino acids per gram of protein of mushrooms are adequate for most FAO amino acid pattern for adults. Multivariate analysis confirmed that protein hydrolysis in processed mushrooms is higher in intestinal phase. Bioaccessibility data for spermidine in A. bisporus is a novelty and increase the value of this food.

Due to the problems raised by the use of animal or microbial recombinant proteases, the use of proteases from vegetable origin is becoming increasingly popular.. Among them, sulfidryl proteases have a special interest. Ficin is an outstanding example of this kind of proteases. This paper aims to be to make a comprehensive review of the recent uses of this enzyme, including for example protein hydrolysis, the production of bioactive peptides and antibodies fragments (researchers point that ficin results are more reproducible than using other proteases), meat tenderization, milk coagulations in cheese making or peptide synthesis. Efforts to get industrial immobilized biocatalysts of the enzyme will be also described. The review shows the huge potential and brilliant prospect that this enzyme can have in the near future. We gratefully recognize the support from the Ministerio de Ciencia e Innovación from Spanish Government (project number CTQ2017-86170-R). The FPU fellowship (Ministerio de Educacion) for Mr. Morellon–Sterling and the fellowship for Mr. Siar from the Algerian Ministry of Higher Education and Scientific Research are also thanked.

The addition of strawberry pulp to ketchup sauces may be an alternative to attend the consumers’ demand for innovative products with functional appeal. Sauces were made using different tomato/strawberry pulp ratios (100:0; 75:25; 50:50; 25:75; 0:100) and characterized for the physicochemical characteristics, bioactive compounds, antioxidant activity, and consumers’ acceptance. The strawberry-enriched ketchup sauces presented higher phenolics, total flavonoids, antioxidant activity, and increased fluidity, and lower browning index, hue angle, chroma, pH, and Bostwick consistency. The antioxidant activities (ABTS and DPPH) were positively correlated with phenolics (r = 0.96 and 0.91) and flavonoids (r = 0.92 and 0.89) contents, respectively. The replacement of up to 50% tomato pulp by strawberry pulp did not change the acidity, flavor, and overall acceptance of the samples. The strawberry pulp has proven to be an effective alternative to increase the concentrations of bioactive compounds while reducing the amount of sugar used in traditional ketchup formulations.

This review presents some of the hottest topics in biotechnological applications: proteases in biocatalysis. Obviously, one of the most relevant areas of application is in the hydrolysis of proteins in food technology, and that has led to a massive use on proteomics. The aim is to identify via peptide maps the different proteins obtained after a specific protease hydrolysis. However, concepts like degradomics are also taking on a more relevant importance in the use and study of proteases and will also be discussed. Other protease applications, as seem in cleaning (detergent development), the pharmaceutical industry, and in fine chemistry, will be analyzed. This review progresses from basic areas such as protease classification to a discussion of the preparation of protease-immobilized biocatalysts, considering the different problems raised by the use of immobilized proteases due to the peculiar features of the substrates, usually large macromolecules. Production of bioactive peptides via limited hydrolysis of proteins will occupy an important place in this review.

Dentre os suplementos dietéticos, os fabricados com proteína de soro de leite apresentam um excelente perfil aminoacídico e caracterizam-se como proteínas de alto valor biológico. No entanto, como resultado da falta de uniformidade, a qualidade nutricional deste tipo de produto é incerta. Assim, este estudo verificou a composição proteica e digestibilidade in vitro desses produtos. Foram adquiridas 12 amostras de suplementos e o teor de proteínas foi determinado pelo método de Kjeldahl. Para a determinação da digestibilidade proteica in vitro foi adotado amedida do percentual de hidrólise de cada amostra, após tratamento com sequência de enzimas pepsina e pancreatina. Ao comparar a composição proteica com os valores declarados nos rótulos dos produtos comercializados observou-se que a maioria das amostras analisadas estava de acordo com o valor declarado. Quanto a digestibilidade proteica in vitro, constatou-se grandes diferenças entre os resultadosdas amostras. Os valores encontrados oscilaram entre 24,2% e 42,2% e estavam abaixo do valor de hidrólise proteica da caseína (48,9%). Esta grande variação pode ser devido às diferentes composições proteicas dos suplementos alimentícios, já que, embora sejam suplementos que trazem em seu rótulo principal o termo whey protein, também veiculam muitas vezes outras proteínas, como proteínas de soja. Além disso, as formulações podem ter em sua composição sucos de frutas ou cacau.Como conclusão, o estudo dos reais teores proteicos, em comparação declarado no rótulo, e mesmo o grau de hidrólise alcançado destas formulações, como indicativo de suas digestibilidades, se mostra cada vez mais importante.

Despite the wide variety of potential applications of lipases within industrial processes, the high cost of production and purification is still their main limiting factor. The aim of this work is to optimize the production of Geotrichum candidum lipase (GCL) using submerged fermentation with a combination of statistical experimental design and surface methodology analysis, in order to give a higher production within a shorter time at the lowest possible cost and easy purification. Cottonseed oil, a low-cost by-product of cotton processing, was used as both an inducer and a carbon source. A maximum lipase activity of 27.17 IUmL-1 was achieved after 30h fermentation in a 5L stirred tank bioreactor under optimal conditions: 2.3% (m/v) of casein peptone, 0.8% (v/v) of cottonseed oil and 0.05% (m/v) of MgSO4 and NaNO3. The lipase purification in a single step by immobilization on PHB particles was verified. The combination of these two steps allowed a significant decrease in this lipase cost of production. Moreover, the produced lipase showed high specificity to hydrolyze long-chain fatty acids with cis-9 double bonds, such as oleic and linoleic acids, having an excellent potential for modifying oils in order to produce different bio-products in industrial applications.

Made available in DSpace on 2018-11-26T17:12:11Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-11-01 Seven different in vitro methods to determine the protein digestibility for chickpea proteins were considered and also the application of these methodologies for calculating PDCAAS (protein digestibility-corrected amino acid score), seeking their correlations with the in vivo methodology. In vitro digestibility of raw and heated samples were determined using pepsin-pancreatin hydrolysis, considering soluble nitrogen via Kjeldahl (ppKJ) and hydrolysed peptide linkages using trinitrobenzenesulfonic acid and o-phthaldialdehyde. In vitro digestibility was also determined using trypsin, chymotrypsin and peptidase (3-Enz) or trypsin, chymotrypsin, peptidase and pronase solution (4-Enz). None of the correlations between in vitro and in vivo digestibilities were significant (at p < 0.0500), but, strong correlations were observed between PDCAAS calculated by in vitro and in vivo results. PDCAAS-ppKJ, PDCAAS-3-Enz and PDCAAS-4-Enz presented the highest correlations with in vivo method, r = 0.9316, 0.9442 and 0.9649 (p < 0.0500), respectively. The use of in vitro methods for calculating PDCAAS may be promising and deserves more discussions. (C) 2016 Elsevier Ltd. All rights reserved. Univ Fed Alfenas, Nutr Fac, 700 Gabriel Monteiro Silva, BR-37130000 Alfenas, MG, Brazil Sao Paulo State Univ, Sch Pharmaceut Sci, Dept Food & Nutr, Araraquara Jau Rd,Km 01, BR-14801902 Araraquara, SP, Brazil Univ Fed Alfenas, Sch Pharmaceut Sci, Food & Med Dept, Gabriel Monteiro da Silva St 700, BR-37130000 Alfenas, MG, Brazil Sao Paulo State Univ, Sch Pharmaceut Sci, Dept Food & Nutr, Araraquara Jau Rd,Km 01, BR-14801902 Araraquara, SP, Brazil

Protein antigenicity can be reduced by enzymatic hydrolysis, which can be performed either by free or immobilized enzyme. The immobilized enzyme is removed from the reaction medium and reused, while the free enzyme must be inactivated to stop the reaction, generally by heating. Here we have shown that hydrolysates produced with free or immobilized Alcalase on glyoxyl-agarose bead presented different physicochemical properties (hydrophilicity profile, molecular mass distribution, surface hydrophobicity) and different levels of residual milk allergens (α-lactalbumin and β-lactoglobulin). Although, under the studied conditions, the hydrolysis with immobilized enzyme did not reduce the residual allergen levels as efficiently as the free enzyme, the results suggest potential applications of immobilized Alcalase for production of hypoallergenic hydrolysates.

FAPEMIG - Fundação de Amparo à Pesquisa do Estado de Minas Gerais Changes in the aqueous solubility of proteins, amino acids, sugars, and other bioactive compounds of the soybean (BRS 257) were studied after different germination times (0, 8, 32, 56, 80, 104 and 176 h). The germination process clearly changed the solubilization of the components, and even antioxidant activities, in aqueous extracts. Significantly (p

Cereal-based diets formulated as semi-purified diets can provide flexibility for researchers, enabling open controlled formulas, besides being cheaper than purified diets. Seeking to widen the researchers’ options in their experimental design developments, we aimed at assaying the chemical score, growth performance and protein utilization of nine semi-purified experimental chows. These diets were formulated at 17.8%, 12% and 8% protein contents, using three variations of ingredients for each one, as follows: casein (C), casein+soybean meal (C+S, 1:1 w/w protein) and casein+soybean meal+corn gluten meal (C+S+G, 1:1:1 w/w protein), without cysteine supplementation and setting casein 17.8% (AIN-93G) as reference diet. The diets C and C+S (17.8%) had the Cys as the limiting amino acid once the addition of gluten turns the Lys as second limiting. All diets had the potential for promoting growth with body mass gain, feed conversion ratio (FCR; chow consumed per body weight gain, average 3.12) and feed efficiency ratio (FER; body weight gain per chow consumed, average 0.3), except for C+S+G 8% (FCR = 6 g; FER = 0.13). These variations were mainly due to the protein amounts independently from the protein ingredient used and could characterize the C+S+G 8% as unable to support growth. For the other parameters (digestibility, net protein ratio, net protein utilization and protein efficiency ratio), there were no relevant differences between the diets. We can conclude that 17.8%, 12% and 8% chows (C and C+S) allowed a proper combination of ingredients from the point of view of palatability, nutrient availability/utilization, metabolic processes, growth performance and feed utilization parameters.

This review intended to give a brief idea of the importance of proteases applications. Processes that involve protein hydrolysis steps find wide ranging utilizations, such as cleaning process, proteomic studies, or food biotechnology process. Many positive effects hoped for with food processing can be achieved by protein hydrolysis using specific proteases, changing nutritional, bioactive and functional properties of food proteins, which include improved digestibility, modifications of sensory quality (such as texture or taste), improvement of antioxidant capability or reduction in allergenic compounds. Protease applications in industrial processes are constantly being introduced and can be advantageous compared to chemical processes, by increasing hydrolysis specificity, product preservation and purity, and reducing environmental impact. Differences in specificity between proteases are very important to take in to consideration as a guide for the choice of protease according to the protein source to be hydrolyzed or predicted products. In this present review, some aspects of the processes that involve protein hydrolysis steps are discussed, especially considering the application of specific proteases as a tool on food biotechnology.

Soymilk was fermented using Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus thermophilus. The soy yoghurt produced was evaluated in comparison to soymilk and commercial milk yoghurt by biological evaluation, rat growth, nitrogen balance assays and microbiological methods. Trypsin inhibitor activity and chemical composition were also determined for all samples. For the soy yoghurt, the growth and nitrogen balance values were not different from the control diet, but the nitrogen balance values were higher than for the soymilk, without significant difference in terms of growth assays. Compared to the commercial yoghurt, Net Protein Ratio and Nitrogen Utilization values were lower, but the Protein Efficiency Ratio, Biological Value and Net Protein Utilization values were equivalent, and for digestibility assays the best results were obtained with the soy yoghurt. The results indicated that soy yoghurt represents a good protein alternative to milk yoghurt and casein. The protein quality of soymilk ...

The chickpea vicilin-like globulin was isolated and chromatographed on Sepharose CL-6B and Sephacryl S-300. The native globulin with a molecular weight of 140 kDa was resolved in Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in seven polypeptide bands in the range of 12.4–67 kDa. The solubility profile of the protein in water and NaCl solutions was typical of a legume globulin. The purified vicilin-like globulin, native and heated, was hydrolyzed by pepsin, trypsin and chymotrypsin. The hydrolysis patterns indicated that the native vicilin-like protein was only partially degraded by the enzymes in comparison with casein. Heating increased its susceptibility to hydrolysis relative to the native form, for all the enzymes. However, the results obtained by the pH-drop method revealed that the in vitro digestibility of the vicilin-like protein was not altered by heating, while 11S-like and total globulins suffered a small increase, indicating that the structural characteristics of storage globulins may be important factors limiting the protein digestion.

Made available in DSpace on 2022-04-28T20:43:09Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-01-01 The amylase from Neurospora crassa is an interesting enzyme, having higher stability than amylase from Aspergillus oryzea under a broad range of pH values. Moreover, the N. crassa enzyme may be immobilized on different supports with good retention of enzyme activity. The best stabilizations were achieved using Eupergit C 250 L or glyoxyl agarose, with which the enzyme remained fully active at 60°C for 24 h while the soluble enzyme remained about 17%. The glyoxyl agarose immobilized enzyme had high thermostability, high optimal temperature (65°C) and broad pH/activity profile, suggesting that this enzyme has potential for food and industrial applications for starch modification. Copyright © Taylor & Francis Group, LLC. Departamento de Bioquimica e Tecnologia Quimica Instituto de Quimica, São Paulo Departamento de Microbiologia de Alimentos Instituto de Fermentaciones Industriales-CSIC, Madrid Departamento de Biocatálisis Instituto de Catálisis-CSIC Madrid Spain Departamento de Alimentos e Nutrição, São Paulo Departamento de Alimentos e Nutrição FCFUNESP, Rodovia Araraquara-Jaú Km 1, 14801 902, Araraquara, São Paulo Instituto de Catalisis Departamento de Biocatálisis Campus UAM, Cantoblanco. 28049 Madrid Departamento de Alimentos e Nutrição FCFUNESP, Rodovia Araraquara-Jaú Km 1, 14801 902, Araraquara, São Paulo

Herbs and spices, excellent sources of phenolic compounds, can be considered potential antioxidant additives. The use of spices must strike a balance between their potential antioxidant capabilities during preparation and the flavor acceptance, in order to avoid rejection of the food. The aimed of this study is to evaluate the influence of different spices and their concentrations on cooked common beans, focusing its potential as antioxidant additives. Onion, parsley, spring onion, laurel and coriander increased the antioxidant activity of preparation when used at 7.96 g of onion, 1.06 g parsley, 3.43 g spring onion, 0.25 g laurel (dry leaves), and 0.43 g coriander/100 g of cooked beans. Besides, these spices concentrations enhance total phenolics and alter the mixture protein digestibility minimally. For garlic samples it was not possible to establish a concentration that increases the antioxidant activity of cooked beans.

A simplified procedure for the preparation of immobilized beta-amylase using non-purified extract from fresh sweet potato tubers is established in this paper, using differently activated agarose supports. Beta-amylase glutaraldehyde derivative was the preparation with best features, presenting improved temperature and pH stability and activity. The possibility of reusing the amylase was also shown, when this immobilized enzyme was fully active for five cycles of use. However, immobilization decreased enzyme activity to around 15%. This seems to be mainly due to diffusion limitations of the starch inside the pores of the biocatalyst particles. A fifteen-fold increase in the Km was noticed, while the decrease of Vmax was only 30% (10.1 U mg−1 protein and 7.03 U mg−1 protein for free and immobilized preparations, respectively)., We would like to thank CNPq and PADC-FCFAr for financial support. We gratefully recognize the support from the Spanish Government, grant CTQ2009-07568.

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T18:41:28Z No. of bitstreams: 1 S1517-83822008000400016.pdf: 1179975 bytes, checksum: 042400e118ea520062326e9e41a151ad (MD5) Made available in DSpace on 2013-08-22T18:41:28Z (GMT). No. of bitstreams: 1 S1517-83822008000400016.pdf: 1179975 bytes, checksum: 042400e118ea520062326e9e41a151ad (MD5) Previous issue date: 2008-12-01 Made available in DSpace on 2013-09-30T18:06:59Z (GMT). No. of bitstreams: 2 S1517-83822008000400016.pdf: 1179975 bytes, checksum: 042400e118ea520062326e9e41a151ad (MD5) S1517-83822008000400016.pdf.txt: 28733 bytes, checksum: 2b9eb0fef4a0dc0c9842e17cf1ac5209 (MD5) Previous issue date: 2008-12-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:56:08Z No. of bitstreams: 2 S1517-83822008000400016.pdf: 1179975 bytes, checksum: 042400e118ea520062326e9e41a151ad (MD5) S1517-83822008000400016.pdf.txt: 28733 bytes, checksum: 2b9eb0fef4a0dc0c9842e17cf1ac5209 (MD5) Made available in DSpace on 2014-05-20T13:56:08Z (GMT). No. of bitstreams: 2 S1517-83822008000400016.pdf: 1179975 bytes, checksum: 042400e118ea520062326e9e41a151ad (MD5) S1517-83822008000400016.pdf.txt: 28733 bytes, checksum: 2b9eb0fef4a0dc0c9842e17cf1ac5209 (MD5) Previous issue date: 2008-12-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Três linhagens de Bacillus sp (BACRP, BACNC- 1 e BACAR) foram isoladas a partir de solo aderido em casca de mandioca. Foram utilizados amido de batata, amido de mandioca, maltodextrina e glicose como fonte de carbono, e temperaturas de crescimento de 25-55ºC, sendo que os três isolados apresentaram maior atividade específica de CGTase quando cultivados com amido de batata a 40ºC. em pH 7,0 os isolados BACRP e BACAR apresentaram atividade específica de 4,0x 10-3 e 2,2x10-3 U/mg prot, respectivamente, quando cultivados em meios acrescidos de 2% de NaCl; em pH 10,0 suas atividades foram de 3,4x10-3 e 3,0x10-3 U/mg prot na mesma concentração de NaCl. Por outro lado, o isolado de BACNC-1 apresentou atividade específica 2,4x10-3 U/mg prot quando cultivado em pH 7,0 acrescido de 1% de NaCl, e em pH 10,0 sua atividade específica foi de 3,4x10-3 U/mg prot sem adição de NaCl. Também foi demonstrada neste trabalho que ciclodextrinas são formadas durante o processo fermentativo, e que a precipitação com acetona ou liofilização seguida de diálise foram eficientes na remoção destas CDs, eliminando sua interferência nos ensaios enzimáticos. A enzima produzida pela cepa BACAR foi purificada parcialmente liberando b-CD como produto da reação. Three strains of Bacillus sp. (BACRP, BACNC-1 and BACAR) were isolated from soil adhered to cassava husk. CGTase specific activity for the three isolated strains was higher when cultivated at 40ºC. Potato starch, cassava starch, maltodextrin and glucose were used as carbon source and growth temperatures varied from 25 to 55ºC. The three isolates presented higher CGTase specific activity when cultivated with potato starch at 40ºC. Isolated BACRP and BACAR presented specific activity of 4.0x10-3 and 2.2x10-3 U/mg prot at pH 7.0, respectively, when cultivated in mediums added with NaCl 2%; at pH 10,0 their activities were of 3.4x10-3 and 3.0x10-3 U/mg prot, respectively, in the same concentration of NaCl. on the other hand, the isolated BACNC-1 presented activity specific of 2.4x10-3 U/mg prot when cultivated at pH 7.0 added of NaCl 1%, and at pH 10.0 the specific activity was of 3.4x10-3 U/mg prot without NaCl addition. This work also showed the presence of cyclodextrins formed during fermentation process and that precipitation with acetone or lyophilization followed by dialysis was efficient at removing CDs (cyclodextrins), thus, eliminating interference in the activity assays. The enzyme produced by the BACAR strain was partially purified and β-CD was liberated as a reaction product. Universidade Estadual Paulista Faculdade de Ciências Farmacêuticas Departamento de Alimentos e Nutrição Universidade Estadual Paulista Instituto de Biociências Departamento de Bioquímica e Microbiologia Universidade Estadual Paulista Faculdade de Ciências Farmacêuticas Departamento de Alimentos e Nutrição Universidade Estadual Paulista Instituto de Biociências Departamento de Bioquímica e Microbiologia FAPESP: 02/09889-6

The aim of this study was to isolate the protein fractions from chickpea, var. IAC-Marrocos, as well as to evaluate its in vivo nutritional protein quality. Among the proteins, albumins showed better nutritional value in the in vivo assays and amino acid contents, despite their higher trypsin inhibitor contents. Trypsin inhibitors were found to be heat labile in all samples, but the digestibility results for unheated and heated flour and albumins suggest that their contents are not very decisive. The PER values for casein (not supplemented) were very similar to those of heated flour and unheated or heated albumin and total globulins. The albumin and glutelin fractions showed the best results for PDCAAS, however, lower than those of casein. Despite the high digestibility of the globulin the very low essential amino acid content lowered its PDCAAS, and it had the lowest values.