RESUMO
Phenolic compounds that are present in pineapple by-products offer many health benefits to the consumer; however, they are unstable to many environmental factors. For this reason, encapsulation is ideal for preserving their beneficial effects. In this work, extracts were obtained by the combined method of solid-state fermentation with Rhizopus oryzae and ultrasound. After this process, the encapsulation process was performed by ionotropic gelation using corn starch, sodium alginate, and Weissella confusa exopolysaccharide as wall material. The encapsulates produced presented a moisture content between 7.10 and 10.45% (w.b), a solubility of 53.06 ± 0.54%, and a wettability of 31.46 ± 2.02 s. The total phenolic content (TPC), antioxidant capacity of DPPH, and ABTS of the encapsulates were also determined, finding 232.55 ± 2.07 mg GAE/g d.m for TPC, 45.64 ± 0.9 µm Trolox/mg GAE for DPPH, and 51.69 ± 1.08 µm Trolox/mg GAE for ABTS. Additionally, ultrahigh performance liquid chromatography (UHPLC) analysis allowed us to identify and quantify six bioactive compounds: rosmarinic acid, caffeic acid, p-coumaric acid, ferulic acid, gallic acid, and quercetin. According to the above, using ionotropic gelation, it was possible to obtain microencapsulates containing bioactive compounds from pineapple peel extracts, which may have applications in the development of functional foods.
RESUMO
The thin-layer chromatography technique (TLC) is a simple and inexpensive analysis commonly used to identify qualitatively the presence of carbohydrates in food samples such as mono- di and oligosaccharides particularly. TLC assay could be improved using image processing software for the semiquantitative determination of this type of compound. In the present work, TLC-image analysis with Silica Gel 60 TLC plates was used for the semiquantitative determination of 6 standards of carbohydrates (glucose, fructose, sucrose, 1-kestose, nystose, and fructofuranosylnystose). Subsequently, the areas of the spots of each compound were determined by digitizing in a conventional office scanner. Then, the segmentation of the images is carried out using software for image processing. The calibration curves were plotted in the Excel software using the average of the areas of the pigmentations obtained in pixels. In this study, the technique of thin-layer chromatography was also used to quantitatively determine the presence of carbohydrates in food samples such as honey, garlic, and onion. Values of determination coefficient (R2) greater than 0.97 in all the calibration curves were obtained. This technique could be useful for detecting carbohydrates (monosaccharides, disaccharides, and oligosaccharides) in analytical assays and food samples without needing specialized analytical equipment. In this work, it was possible to determine the concentration of carbohydrates in samples of garlic and onion that showed the presence of prebiotic carbohydrates in addition to sucrose, glucose, and fructose.
RESUMO
Cultivable halophilic microorganisms were isolated and identified from saline and alkaline-sodic soils: Cuatro Cienegas, Sayula and San Marcos lakes. Physicochemical characteristics of soils were determined to understand the relationship between those and the microorganisms isolated. The Cuatro Cienegas soils had a neutral pH, EC of 2.3-8 dS cm-1, classified as moderately saline. Whereas, the soils from Sayula and San Marcos lakes, had an alkaline pH, EC 15 to 65 dS m-1, typical of saline-sodic. We identified 23 cultivable halophilic bacteria using 16s rDNA, being Halobacillus sp., Marinococcus sp., and Alkalibacillus sp. the predominant genus by culture dependent approach. We found a correlation between the soils anion and cation content with the occurrence of different genus of halophilic bacteria in each studied site. Alkalibacillus sp. was predominant in Sayula and San Marcos lakes and was related to the high Na+ content; while Bacillus sp. and Halobacillus sp. were predominant in Cuatro Cienegas, their occurrence was related to a high content of Ca2+, Mg2+, and SO4 2-.
RESUMO
Abstract Hamelia patens, is a plant traditionally used to treat a variety of conditions among the Huastec people of Mexico. The objective of this study is to characterize the phenolic content and critically examine the antimicrobial activity of leaf extracts H. patens, obtained by maceration, Soxhlet and percolation, using ethanol as 70% solvent. Phenolic compounds are characterized by liquid chromatography, coupled to a High Resolution Mass Spectrometry, and the antimicrobial activity was studied from the inhibitory effect of each extract for Escherichia coli, Staphylococcus aureus, Salmonella typhi and S. paratyphi, and by the Minimum Bactericidal Concentration, the percentage of activity and the Index of Bacterial Susceptibility of each extract. The phenolic compound identified in different concentrations in the three extracts was epicatechin. The extracts obtained by the three methods had antimicrobial activity, however, there was no significant difference (p < 0.05) between the Minimum Bactericidal Concentration of the extracts obtained by maceration, percolation and Soxhlet. The results of this study contribute to the body of knowledge on the use of extracts in controlling microorganisms with natural antimicrobials.
Assuntos
Fenóis/isolamento & purificação , Fenóis/farmacocinética , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacocinética , Hamelia/química , Fracionamento Químico/métodos , Antibacterianos/isolamento & purificação , Antibacterianos/farmacologia , Fenóis/química , Staphylococcus aureus/efeitos dos fármacos , Extratos Vegetais/química , Testes de Sensibilidade Microbiana , Escherichia coli/efeitos dos fármacos , México , Antibacterianos/químicaRESUMO
Hamelia patens, is a plant traditionally used to treat a variety of conditions among the Huastec people of Mexico. The objective of this study is to characterize the phenolic content and critically examine the antimicrobial activity of leaf extracts H. patens, obtained by maceration, Soxhlet and percolation, using ethanol as 70% solvent. Phenolic compounds are characterized by liquid chromatography, coupled to a High Resolution Mass Spectrometry, and the antimicrobial activity was studied from the inhibitory effect of each extract for Escherichia coli, Staphylococcus aureus, Salmonella typhi and S. paratyphi, and by the Minimum Bactericidal Concentration, the percentage of activity and the Index of Bacterial Susceptibility of each extract. The phenolic compound identified in different concentrations in the three extracts was epicatechin. The extracts obtained by the three methods had antimicrobial activity, however, there was no significant difference (p < 0.05) between the Minimum Bactericidal Concentration of the extracts obtained by maceration, percolation and Soxhlet. The results of this study contribute to the body of knowledge on the use of extracts in controlling microorganisms with natural antimicrobials.(AU)
Assuntos
Hamelia , Extratos Vegetais/administração & dosagem , Extratos Vegetais/uso terapêutico , Antibacterianos/farmacologia , Compostos Fenólicos/análise , Cromatografia Líquida de Alta Pressão/métodosRESUMO
Hamelia patens, is a plant traditionally used to treat a variety of conditions among the Huastec people of Mexico. The objective of this study is to characterize the phenolic content and critically examine the antimicrobial activity of leaf extracts H. patens, obtained by maceration, Soxhlet and percolation, using ethanol as 70% solvent. Phenolic compounds are characterized by liquid chromatography, coupled to a High Resolution Mass Spectrometry, and the antimicrobial activity was studied from the inhibitory effect of each extract for Escherichia coli, Staphylococcus aureus, Salmonella typhi and S. paratyphi, and by the Minimum Bactericidal Concentration, the percentage of activity and the Index of Bacterial Susceptibility of each extract. The phenolic compound identified in different concentrations in the three extracts was epicatechin. The extracts obtained by the three methods had antimicrobial activity, however, there was no significant difference (p<0.05) between the Minimum Bactericidal Concentration of the extracts obtained by maceration, percolation and Soxhlet. The results of this study contribute to the body of knowledge on the use of extracts in controlling microorganisms with natural antimicrobials.
Assuntos
Antibacterianos/isolamento & purificação , Antibacterianos/farmacologia , Fracionamento Químico/métodos , Hamelia/química , Fenóis/isolamento & purificação , Fenóis/farmacocinética , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacologia , Antibacterianos/química , Escherichia coli/efeitos dos fármacos , México , Testes de Sensibilidade Microbiana , Fenóis/química , Extratos Vegetais/química , Staphylococcus aureus/efeitos dos fármacosRESUMO
OBJECTIVE: To determinate the recovery of total polyphenolic compounds content, in vitro antioxidant activity and HPLC/ESI/MS characterization of extract from Nephelium lappaceum L. (Mexican rambutan). METHODS: The rambutan husk extract was obtained by aqueous extraction and a polyphenolic fraction was recovered using Amberlite XAD-16. The total polyphenolic compounds content was determined by the Folin Ciocalteu and butanol-HCI methods. In vitro antioxidant activity was performed using ABTS and ferric reducing antioxidant power methods. RESULTS: Mexican rambutan husk showed a total polyphenolic content of 582 mg/g and an evident antioxidant activity by ABTS and ferric reducing antioxidant power analysis. The HPLC/ESI/MS assay allowed the identification of 13 compounds, most of which belong to ellagitannins. Geraniin, corilagin and ellagic acid were present in the sample; the mineral composition was also evaluated. CONCLUSIONS: Rambutan husk cultivated in Mexico is a promising source for the recovery of added value bioactive compounds with antioxidant activity, which have potential applications as bioactive antioxidant agents for the treatment of diseases.
RESUMO
Fungal hydrolysis of ellagitannins produces hexahydroxydiphenic acid, which is considered an intermediate molecule in ellagic acid release. Ellagic acid has important and desirable beneficial health properties. The aim of this work was to identify the effect of different sources of ellagitannins on the efficiency of ellagic acid release by Aspergillus niger. Three strains of A. niger (GH1, PSH and HT4) were assessed for ellagic acid release from different polyphenol sources: cranberry, creosote bush, and pomegranate used as substrate. Polyurethane foam was used as support for solid-state culture in column reactors. Ellagitannase activity was measured for each of the treatments. Ellagic acid was quantified by high performance liquid chromatography. When pomegranate polyphenols were used, a maximum value of ellagic acid (350.21 mg/g) was reached with A. niger HT4 in solid-state culture. The highest amount of ellagitannase (5176.81 U/l) was obtained at 8 h of culture when cranberry polyphenols and strain A. niger PSH were used. Results demonstrated the effect of different polyphenol sources and A. niger strains on ellagic acid release. It was observed that the best source for releasing ellagic acid was pomegranate polyphenols and A. niger HT4 strain, which has the ability to degrade these compounds for obtaining a potent bioactive molecule such as ellagic acid.
La hidrólisis fúngica de los elagitaninos produce ácido hexahidroxidifénico, considerado como una molécula intermedia en la liberación de ácido elágico. El ácido elágico tiene importantes y deseables propiedades benéficas para la salud humana. El objetivo de este trabajo fue identificar el efecto de la fuente de elagitaninos sobre la eficiente liberación de ácido elágico por Aspergillus niger. La liberación de ácido elágico se realizó con tres cepas de A. niger (GH1, PSH y HT4) en presencia de diferentes fuentes de polifenoles (arándano, gobernadora y granada), usadas como sustrato. Se empleó espuma de poliuretano como soporte para el cultivo en estado sólido en reactores en columna. Se midió la actividad elagitanasa a cada uno de los tratamientos. El ácido elágico liberado se cuantificó por cromatografía líquida de alta resolución. Cuando se utilizaron los polifenoles de granada, se alcanzó un valor máximo de 350,21 mg/g de ácido elágico con A. niger HT4 en cultivo en estado sólido. La mayor actividad elagitanasa (5176.81 U/l) se obtuvo a 8 h de cultivo cuando se usaron los polifenoles de arándano como sustrato y A. niger PSH. Los resultados demostraron el efecto que tiene la fuente de polifenoles y la cepa de A. niger en la liberación de ácido elágico. Se observó que la mejor fuente para la liberación de ácido elágico fueron los polifenoles de granada y que la cepa A. niger HT4 posee la habilidad de degradar estos compuestos para la obtención de potentes moléculas bioactivas, como el ácido elágico.
Assuntos
Aspergillus niger/isolamento & purificação , Ácido Elágico/análise , Polifenóis/análise , Aspergillus niger/fisiologia , Cromatografia Líquida de Alta Pressão/métodosRESUMO
Fungal hydrolysis of ellagitannins produces hexahydroxydiphenic acid, which is considered an intermediate molecule in ellagic acid release. Ellagic acid has important and desirable beneficial health properties. The aim of this work was to identify the effect of different sources of ellagitannins on the efficiency of ellagic acid release by Aspergillus niger. Three strains of A. niger (GH1, PSH and HT4) were assessed for ellagic acid release from different polyphenol sources: cranberry, creosote bush, and pomegranate used as substrate. Polyurethane foam was used as support for solid-state culture in column reactors. Ellagitannase activity was measured for each of the treatments. Ellagic acid was quantified by high performance liquid chromatography. When pomegranate polyphenols were used, a maximum value of ellagic acid (350.21 mg/g) was reached with A. niger HT4 in solid-state culture. The highest amount of ellagitannase (5176.81 U/l) was obtained at 8h of culture when cranberry polyphenols and strain A. niger PSH were used. Results demonstrated the effect of different polyphenol sources and A. niger strains on ellagic acid release. It was observed that the best source for releasing ellagic acid was pomegranate polyphenols and A. niger HT4 strain, which has the ability to degrade these compounds for obtaining a potent bioactive molecule such as ellagic acid.
Assuntos
Aspergillus niger/efeitos dos fármacos , Aspergillus niger/metabolismo , Ácido Elágico/metabolismo , Taninos Hidrolisáveis/farmacologia , Extratos Vegetais/farmacologia , Polifenóis/farmacologia , Larrea , Lythraceae , Vaccinium macrocarponRESUMO
Tannin acyl hydrolases, or tannases (EC 3.1.1.20), are enzymes with potential biotechnological applications. In this work, we describe the gene and amino acid sequences of the tannase from Aspergillus niger GH1. In addition, we engineered Pichia pastoris strains to produce and secrete the enzyme, and the produced tannase was characterized biochemically. The nucleotide sequence of mature tannase had a length of 1,686 bp, and encodes a protein of 562 amino acids. A molecular model of mature A. niger GH1 tannase showed the presence of two structural domains, one with an α/ß-hydrolase fold and one lid domain that covers the catalytic site, likely being residues Ser-196, Asp-448, and His-494 the putative catalytic triad, which are connected by a disulfide bond between the neighboring cysteines, Cys-195 and Cys-495. A 120-ml shake flask culture with a constructed recombinant P. pastoris strain showed extracellular tannase activity at 48 h induction of 0.57 U/ml. The produced tannase was N-glycosylated, consisted of two subunits, likely linked by a disulfide bond, and had an optimum pH of 5.0 and optimum temperature of 20 °C. These biochemical properties differed from those of native A. niger GH1 tannase. The recombinant tannase could be suitable for food and beverage applications.
Assuntos
Aspergillus niger/enzimologia , Hidrolases de Éster Carboxílico/genética , Hidrolases de Éster Carboxílico/metabolismo , Pichia/genética , Sequência de Aminoácidos , Aspergillus niger/genética , Hidrolases de Éster Carboxílico/química , Domínio Catalítico , Proteínas Fúngicas/química , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Glicosilação , Modelos Moleculares , Pichia/metabolismo , Estrutura Secundária de Proteína , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismoRESUMO
ß-Fructofuranosidases or invertases (EC 3.2.1.26) are enzymes that are widely used in the food industry, where fructose is preferred over sucrose, because it is sweeter and does not crystallize easily. Since Aspergillus niger GH1, an xerophilic fungus from the Mexican semi-desert, has been reported to be an invertase producer, and because of the need for new enzymes with biotechnological applications, in this work, we describe the gene and amino acid sequence of the invertase from A. niger GH1, and the use of a synthetic gene to produce the enzyme in the methylotrophic yeast Pichia pastoris. In addition, the produced invertase was characterized biochemically. The sequence of the invertase gene had a length of 1770 bp without introns, encodes a protein of 589 amino acids, and presented an identity of 93% and 97% with invertases from Aspergillus kawachi IFO 4308 and A. niger B60, respectively. A 4.2 L culture with the constructed recombinant P. pastoris strain showed an extracellular and periplasmic invertase production at 72 h induction of 498 and 3776 invertase units (U), respectively, which corresponds to 1018 U/L of culture medium. The invertase produced had an optimum pH of 5.0, optimum temperature of 60 °C, and specific activity of 3389 U/mg protein, and after storage for 96 h at 4 °C showed 93.7% of its activity. This invertase could be suitable for producing inverted sugar used in the food industry.
Assuntos
Aspergillus niger/genética , Proteínas Fúngicas/genética , beta-Frutofuranosidase/genética , Sequência de Aminoácidos , Aspergillus niger/enzimologia , Sequência de Bases , Sistema Livre de Células , Clonagem Molecular , Líquido Extracelular/enzimologia , Frutose/biossíntese , Proteínas Fúngicas/isolamento & purificação , Proteínas Fúngicas/metabolismo , Genes Sintéticos , Glucose/biossíntese , Concentração de Íons de Hidrogênio , Microbiologia Industrial/métodos , Periplasma/enzimologia , Pichia , Estabilidade Proteica , Proteínas Recombinantes de Fusão/metabolismo , Homologia de Sequência , Sacarose/metabolismo , Temperatura , beta-Frutofuranosidase/isolamento & purificação , beta-Frutofuranosidase/metabolismoRESUMO
The combined effects of pH and temperature on red pigment production and fungal morphology were evaluated in a submerged culture of Penicillium purpurogenum GH2, using Czapek-Dox media with d-xylose as a carbon source. An experimental design with a factorial fix was used: three pH values (5, 7, and 9) and two temperature levels (24 and 34 °C) were evaluated. The highest production of red pigment (2.46 g/L) was reached with a pH value of 5 and a temperature of 24 °C. Biomass and red pigment production were not directly associated. This study demonstrates that P. purpurogenum GH2 produces a pigment of potential interest to the food industry. It also shows the feasibility of producing and obtaining natural water-soluble pigments for potential use in food industries. A strong combined effect (p<0.05) of pH and temperature was associated with maximal red pigment production (2.46 g/L).
Assuntos
Penicillium/química , Penicillium/metabolismo , Pigmentos Biológicos/biossíntese , Adaptação Fisiológica/fisiologia , Tamanho Celular , Concentração de Íons de Hidrogênio , Penicillium/citologia , TemperaturaRESUMO
Tannase is an inducible enzyme with important applications in the food and pharmaceutical industries. This enzyme was produced by the fungus Aspergillus niger GH1 under solid-state fermentation using polyurethane foam as solid support and tannic acid as sole carbon source and tannase inducer. Physicochemical properties of A. niger tannase were characterized, and the kinetic and thermodynamics parameters on methyl gallate hydrolysis were evaluated. The enzyme was stable in a pH range of 2-8 and a functional temperature range of 25-65 °C. The highest k(cat) value was 2,611.10 s(-1) at 65 °C. Tannase had more affinity for methyl gallate at 45 °C with a K(M) value of 1.82 mM and an efficiency of hydrolysis (k(cat)/K(M)) of 330.01 s(-1) mM(-1). The lowest E(a) value was found to be 21.38 kJ/mol at 4.4 mM of methyl gallate. The lowest free energy of Gibbs (ΔG) and enthalpy (ΔH) were found to be 64.86 and 18.56 kJ/mol, respectively. Entropy (ΔS) was -0.22 kJ/mol K. Results suggest that the A. niger GH1 tannase is an attractive enzyme for industrial applications due its catalytic and thermodynamical properties.
Assuntos
Aspergillus niger/enzimologia , Hidrolases de Éster Carboxílico/química , Proteínas Fúngicas/química , Aspergillus niger/crescimento & desenvolvimento , Técnicas de Cultura Celular por Lotes/instrumentação , Biocatálise , Hidrolases de Éster Carboxílico/metabolismo , Estabilidade Enzimática , Fermentação , Proteínas Fúngicas/metabolismo , Concentração de Íons de Hidrogênio , Cinética , Poliuretanos/análiseRESUMO
Según la definición otorgada por la Organización de las Naciones Unidas para la Agricultura y la Alimentación (FAO), los prebióticos son componentes no vivos de los alimentos que confieren un beneficio saludable al huésped, asociado con la modulación de la microbiota. Los compuestos prebióticos incluyen oligosacáridos (fructooligosácaridos, galactooligosacáridos, xylooligosacáridos, pecticoligosacáridos), lactosacarosa, azúcares-alcoholes, glucooligosacáridos, levanos o fructanos, almidón resistente, xylosacáridos, entre otros. Los procesos de recuperación, síntesis y/o purificación son específicos para cada grupo. El metabolismo de la microflora produce la formación de gases como H2, CO2 y CH4, y compuestos orgánicos (ácidos grasos de cadena corta y etanol) como producto de la fermentación de prebióticos. Los efectos observados por acción de los prebióticos impactan en la salud del consumidor y pueden manifestarse de forma localizada como aumento de la masa fecal, de la absorción colónica de algunos minerales y de la síntesis de ácido fólico. También, pueden observarse de forma sistémica con la disminución de colesterol, triglicéridos, amonio, urea, entre otras actividades. Una de las aplicaciones con mayor potencial de los estudios de pre y probióticos, es la formulación de alimentos simbióticos; así mismo, los prebióticos prometen cumplir con las necesidades actuales de los consumidores, quienes demandan alimentos funcionales.
According to the definition given by the United Nations Food and Agriculture Organization (FAO), prebiotics are non live components of food that grant a health benefit to the host, associated with the modulation the intestinal microbiota. Prebiotic compounds include, among others, oligosaccharides (fructooligosaccharides, galactooligosaccharides, xylooligosaccharides, pecticooligosaccharides), lactosaccharose, sugar-alcohols, glucooligosccharides, levans or fructans, resistant starch, and xylosaccharides. The recovery, synthesis and/or purification processes are specific for each group. Microflora metabolism produces formation of gases such as H2, CO2 and CH4, and organic compounds (short chain fatty acids and ethanol) as products of prebiotic fermentation. The effects on the health of the consumer produced by the action of the prebiotics can be manifested locally, as by an increase of fecal mass, colon absorption of some minerals, and folic acid synthesis. They can also be observed systemically, as by a decrease of cholesterol, triglycerides, ammonium, and urea, among other activities. One of the applications with the greatest potential in the study of pre and probiotics is the formulation of symbiotic food; probiotics also promise to fulfill the present needs of comsumers, who demand functional food.
RESUMO
Significant differences on structure, stability, and catalytic properties of tannase were found when this enzyme was produced under solid-state and submerged fermentations (SSF and SmF) by Aspergillus niger. The specific activity was 5.5 times higher on SSF than in SmF. Significant differences in isoelectric points of tannases were found. The pH optima for both types of enzyme was found at 6 and the pH stability of SSF and SmF tannase were at 6 and 5-8, respectively. The optimal temperature range was from 50 to 60 °C for SmF tannase and 60 °C for SSF tannase, and both enzyme types showed tolerance to high temperatures (60-70 °C). The SSF tannase showed a major specificity for methyl gallate substrate while SmF tannase for tannic acid. All metal ions tested, had an activity inhibition from 30-46% on SSF tannase. SDS-PAGE analysis as well as gel localization studies of both SSF and SmF purified tannases showed a single band with a molecular weight of 102 and 105 kDa, respectively. Different levels of glycosylation were found among SSF and SmF purified tannases. This is the first report about structural differences among tannase produced under SSF and SmF and this study provides basis for explanation of the stability and catalytic differences observed previously for this two tannase types.
Assuntos
Aspergillus niger/enzimologia , Aspergillus niger/metabolismo , Hidrolases de Éster Carboxílico/metabolismo , Eletroforese em Gel de Poliacrilamida , Fermentação/fisiologia , Concentração de Íons de Hidrogênio , TemperaturaRESUMO
The influence of the physical structure of polyurethane matrix as a support in a solid state culture in tannase production and gallic acid accumulation by Aspergillus niger Aa-20 was evaluated. Three different polyurethane matrices were used as the support: continuous, semi-discontinuous and discontinuous. The highest tannase production at 2479.59 U/L during the first 12 h of culture was obtained using the discontinuous matrix. The gallic acid was accumulated at 7.64 g/L at the discontinuous matrix. The results show that the discontinuous matrix of polyurethane is better for tannase production and gallic acid accumulation in a solid state culture bioprocess than the continuous and semi-discontinuous matrices.
Assuntos
Aspergillus niger/metabolismo , Reatores Biológicos/microbiologia , Hidrolases de Éster Carboxílico/metabolismo , Técnicas de Cultura de Células/métodos , Ácido Gálico/metabolismo , Poliuretanos/químicaRESUMO
Pectinesterase was extracted from potato alpha cultivar, purified and partially characterized The used protocol resulted in a 58.8-fold purification (51 850.2 units/mg protein) with 15.5 percent recovery of pectinesterase activity. The purified enzyme had a molecular weight of 27 kDa and its isoelectric point was around 4.5 with pH and temperature optima of 8.0 and 60°C, respectively. The purified enzyme had a single symmetric peak of specific activity after chromatographic steps. The homogeneity of the purified pectinesterase was confirmed by gel filtration and polyacrylamide electrophoresis gel.
A pectinesterase foi extraída da batata (cultivar do alfa), purificada e parcialmente caracterizada. O protocolo usado levou a uma proteína purificada 58,8 vezes (51 850,2 units/mg da proteína) com uma recuperação de 15,5 por cento da atividade da proteína. A enzima purificada apresentou um peso molecular de 27 kDa e seu ponto isoelétrico foi ao redor 4,5. A pectinesterase exibiu pH e temperatura ótimos de respectivamente 8,0 e 60°C. A enzima purificada apresentou um único pico simétrico de atividade específica após as etapas de cromatografia. A homogeneidade da pectinesterase purificada foi confirmada por filtração em gel e por eletroforese em gel de poliacrilamida.