924 resultados para Different Proteinase Activity
Resumo:
The Brazilian Savannah, known as "Cerrado," has an extensive biodiversity, but it is under explored. Among the native vegetables is the jatobá-do-cerrado (Hymenaea stigonocarpa Mart.), a legume with great potential for exploration for its content of dietary fiber. Legumes are an important source of nutrient compounds, such as phenolic compounds and vitamins that have antioxidant properties. This study aimed at determining the chemical composition and antioxidant activity of the jatobá flour. The jatobá flour showed high fiber content (insoluble and soluble fiber 47.8 and 12.8 g.100 g- 1, respectively), significant amounts of carotenoids such as beta-carotene and lutein, and some minerals such as calcium: 145 mg.100 g- 1, magnesium: 125 mg.100 g- 1, and potassium: 1352 mg.100 g- 1. The jatobá flour extracted with different solvents (water, methanol, and acetone) exhibited antioxidant activity by the DPPH, FRAP, and ORAC methods. The solvent used in the extraction affected the total phenolic content and antioxidant activity. Acetone extraction produced the best results. Therefore, the jatobá flour is an ingredient that can be used to develop new products with properties that promote health.
Resumo:
Sweet potato is an important staple, and it is mainly known for its contribution of β-carotene in human diet. The effects of cultivar and habitat on this pigment and other nutritional characteristics of the crop still require investigation. In this study, three locally bred cultivars of sweet potato, two of which are orange-fleshed, were grown in three different agro-ecological areas to determine soluble sugar content, β-carotene, and total antioxidants of roots. In addition antioxidant activity, total carotenoids, and chlorophyll content were determined in edible leaves. Reducing sugars, β-carotene, total antioxidants capacity, total carotenoids, and chlorophyll content were significantly affected by environmental conditions. The location at lower altitude and closer to the coastline showed high evapotranspiration, thus reducing sugar content, antioxidant activity, and phytonutrients in both storage roots and leaves. Absence of water stress in agro-ecological locations further inland and at higher altitudes was associated with an increase in these compounds. Free radical scavenging activity of DPPH was higher in the storage roots (610.49 µmoles TE/100g) than in the leaves (426.06 µmoles TE/100g); nevertheless, opposite results were found for the ferric ion reducing activity (FRAP). The deep orange-fleshed cultivar A45 contained high β-carotene (15 mg/100g), which is enough to meet RDA for vitamin A. There is evidence of agro-ecological effect on sweet potato nutritional value.
Resumo:
AbstractAscorbic acid, carotenoids and polyphenols stand out among the orange juice natural antioxidants. The winemaking process affects their bioavailability and bioactivity. Antioxidant activities (AA) were estimated in different process conditions to asses those properties. The AA and their correlation with ascorbic acid, total phenolics and carotenoids content were calculated. The variables and levels analyzed were: pasteurized and natural must (PJ and NJ), pH 3.5 and 4.0 and fermentation temperatures at 10°C and 20°C. Statistically significant differences (α=0.05) were found among bioactive compounds concentrations. Antioxidant compounds concentration was higher in raw material than in orange wine. Juice pasteurization caused the major losses while subsequent vinification affects them to a lesser extent. Highest antioxidants retention was measured in wines from JN fermented at pH 3.5 and 10 °C (JN-3.5-10) followed by wines from JP and fermented at the same conditions (JP-3.5-10). AA determined by DPPH showed a positive and close correlation with FRAP, while ABTS showed a low correlation with former assays. Juice pasteurization process and fermentation temperature influenced bioactive compound reduction which correlates with the AA variation.
Resumo:
Abstract The changes in flaxseed constituents at different stages of maturity are reported. The physical properties and antioxidant activity of flaxseed oil during flaxseed development have been evaluated. Continuous decrease in total polyphenol content during flaxseed development. All the results showed no significant differences between HPLC-MS and TLC for quantitative determination of phospholipids classes. The fatty acid compositions of individual phospholipids were also reported. The antioxidant activity of oilseed was assessed by means of 2, 2-diphenyl-1-picryl-hydrazyl (DPPH) radical scavenging assay. The test demonstrated that the antioxidant activity of the flaxseed oil was found to be harvesting time-dependant.
Resumo:
Abstract The objective of this work was to study the effect of enzymatic hydrolysis of black bean protein concentrate using different enzymes. Bean proteins were extracted and hydrolyzed over a period of 120 min using the enzymes pepsin or alcalase. The protein hydrolysates’ molecular weight was assayed by electrophoresis and the antioxidant activity was evaluated by the capturing methods of free radicals ABTS●+ and DPPH. Electrophoretic results showed that the bands above 50 kDa disappeared, when the beans protein was subjected to hydrolysis with pepsin. The bean protein hydrolysate obtained by hydrolysis with alcalase enzyme, showed higher antioxidant activity for inhibition of the radical ABTS●+. However, the hydrolysates obtained by hydrolysis with pepsin had higher antioxidant activity for inhibition of the radical DPPH. The use of pepsin and alcalase enzymes, under the same reaction time, produced black bean protein hydrolysates with different molecular weight profiles and superior antioxidant activity than the native bean protein.
Resumo:
Abstract The research work aimed at investigating the effect of pre-harvest gibberellic acid (GA3) treatment on the quality of ‘Obilnaja’ and ‘Black Star’ Japanese plum varieties. GA3 was sprayed onto the trees during the fruit color break at 0, 25, 50, 75, and 100 ppm concentrations. After pre-cooling, the plums were placed in modified atmosphere packages and exposed to the following conditions as follows: short storage-transportation (ST) [20 days at 2 °C and 90% relative humidity (RH)]; distribution center (DC) (5 days at 6 °C and 80% RH), and shelf life conditions (SL) (2 days at 20 °C and 70% RH). Pre-harvest GA3 treatments increased the fruit weight and size. Treatment of GA3 at 50, 75, and 100 ppm increased the fruit flesh firmness and total soluble substances (TSS) values in both the plum varieties during storage, transport, and marketing; it also limited the weight loss during the marketing process. Treatment of GA3 had no significant effects on the color, titratable acidity (TA), and the total phenolic and antioxidant activity values of plums. Pre-harvest GA3 treatment at 50 ppm GA3 can be thus recommended for both the plum varieties due to its effect on the fruit quality.
Resumo:
Abstract The objective of this work was to evaluate the antioxidant activity of protein hydrolysates obtained by the enzymatic hydrolysis of okara using an endopeptidase (Alcalase) and exopeptidase (Flavourzyme). The reaction was monitored by the pH-stat procedure in which five aliquots were collected during the hydrolysis by each enzyme, corresponding to different degrees of hydrolysis (DH). The antioxidant activities of the aliquots were evaluated by the ABTS, DPPH and FRAP methods. For the hydrolysates obtained using Alcalase, the antioxidant activities increased from: 68.6 to 99.5% (ABTS), 14.5 to 17.7% (DPPH) and 222.6 to 684.9 µM Trolox (FRAP), when the DH varied from 0 to 33.6%. With respect to Flavourzyme, the results were: 67.2 to 88.2% (ABTS), 9.5 to 18.5% (DPPH) and 168.0 to 360.3 µM Trolox (FRAP), when the DH increased up to 5.8%. The results showed that the protein hydrolysates had antioxidant capacities, which were influenced by the degree of hydrolysis and the type of enzyme.
Resumo:
Currently, the highest interest with respect to the assessment of seed physiological quality is to obtain reliable results in a relatively short period of time. This initiative allows for prompt decisions during different phases of seed production primarily after physiological maturity. This research was performed to verify the efficiency and rapidity of the method of Pettenkofer to determine the respiratory activity and to differentiate vigor levels of soybean seed lots. Three lots of soybean seeds cv. 8000 were used. Seed performance was determined by respiratory activity, compared to the following tests: standard germination, germination first count, electrical conductivity, seedling emergence, seedling shoot and root length and total dry mass. Results ranked seed lots according to defferences in physiological quality. Seed imbibition and conditioning period in Pettenkofer's equipment were enough to detect differences in vigor among seed lots, showing that the determination of the respiratory activity is a promising procedure to identify differences in vigor levels among soybean seed lots.
Resumo:
The objective of this study was to evaluate physiological quality, content, and activity of antioxidants, in soybean seeds subjected to accelerated aging during different periods. Seeds of cultivars BRS 258, BRS 262 and BRS 268, subjected to accelerated aging during 12, 24, 36 and 48 hours and non-aged seeds were used. After each aging period, the seed were evaluated by tests of: germination; first count and tetrazolium. The total of phenolic compounds, total flavonoides, total of isoflavones, and activity for eliminating ABTS●+ radicals were quantified. There were differences among cultivars according to vigor and viability only after seeds were aged. Cultivars BRS 158 and BRS 268 have shown better seed physiological quality in each aging period; however, not presenting higher amounts of isoflavones and efficiency in removing free radicals. For all cultivars, the values for total of phenolic compounds, as well as total of flavonoids have shown quadratic positive behavior; the values for isoflavones remained constant and the vigor and viability showed contrary trend to activity of antioxidant agents.
Resumo:
Green mould is a serious disease of commercially grown mushrooms, the causal agent being attributed to the filamentous soil fungus Triclzodenna aggressivum f. aggressivu11l and T. aggressivum f. ellropaellm. Found worldwide, and capable of devastating crops, this disease has caused millions of dollars in lost revenue within the mushroom industry. One mechanism used by TricllOdenlla spp. in the antagonism of other fungi, is the secretion of lytic enzymes such as chitinases, which actively degrade a host's cell wall. Therefore, the intent of this study was to examine the production of chitinase enzymes during the host-parasite interaction of Agaricus bisporus (commercial mushroom) and Triclzodemza aggressivum, focusing specifically on chitinase involvement in the differential resistance of white, off-white, and brown commercial mushroom strains. Chitinases isolated from cultures of A. bisporus and T. aggressivu11l grown together and separately, were identified following native PAGE, and analysis of fluorescence based on specific enzymatic cleavage of 4-methylumbelliferyl glucoside substrates. Results indicate that the interaction between T. aggressivulll and A. bisporus involves a complex enzyme battle. It was determined that T. aggressivum produces a number of chitinases that appear to correlate to those isolated in previous studies using biocontrol strains of T. Izarziallilm. A 122 kDa N-acetylglucosaminidase of T. aggressivu11l revealed the highest and most variable activity, and is therefore believed to be an important predictor of antifungal activity. Furthermore, results indicate that brown strain resistance of mushrooms may be related to high levels of a 96 kDa N-acetylglucosaminidase, which showed elevated activity in both solitary and dual cultures with T. aggressivum. Overall, each host-parasite combination produced unique enzyme profiles, with the majority of the differences seen between day 0 and day 6 for the extracellular chitinases. Therefore, it was concluded that the antagonistic behaviour of T. aggressivli1ll does not involve a typical response, always producing the same types and levels of enzymes, but that mycoparasitism, specifically in the form of chitinase production, may be induced and regulated based on the host presented.
Resumo:
Physical inactivity poses a huge burden on Canada's health care system and is detrimental to the health of Canadians (Katzmarzyk & Janssen, 2004). Walking is a viable option for individuals to become physically active on a daily basis and is in fact the most commonly reported leisure time physical activity. It has been associated with many health benefits including weight loss/weight control, reduced risk of coronary artery disease and diabetes, lowered blood pressure, and improved psychological wellbeing (Brisson & Tudor-Locke, 2004). Specifically, individuals' stage of change, selfefficacy and health related quality of life (HRQL) are three psychological constructs that can be greatly improved with increased physical activity (Dishman, 1991; Penedo & Dahn, 2005; Poag & McAuley, 1992). Public health physical activity recommendations exist but many individuals find these difficult to meet due to overly busy lifestyles (Public Health Agency of Canada, 2003). Pedometers are inexpensive devices that can monitor individual bouts of walking so that the incorporation of physical activity into one's daily life is more plausible. They are also excellent tools for motivation, goalsetting, and immediate feedback (Brisson & Tudor-Locke, 2004). Since many people spend a large proportion of their time at their places of employment, workplaces have begun to be a common site for the development of physical activity interventions. These programs have been growing in popUlarity and have shown numerous benefits for both employees and employers (Voit, 2001). The purpose of the current study was to implement and evaluate the use of a pedometer-based physical activity intervention incorporating goal-setting and physical activity logs in a workplace setting, and to examine the relationship between different types of self-efficacy (task, barrier, and scheduling) and different phases of the intervention. Twenty male participants from a local steel manufacturing plant who exhibited health risk factors (e.g. hypertension, diabetes, etc.) were assigned to one of two groups (group A or group B). All participants were asked to wear pedometers on their waists, record their daily steps, set goals that were outlined on a step-tracking sheet (detennined by their baseline number of steps), and keep track of their work days, wakelbed time, sedentary time, and time spent doing other physical activity. Group A began the intervention immediately following the baseline measures, whereas group B continued with their regular routine for 4 weeks before beginning. Physiological measures (height, weight, blood pressure, relative body fat, waist and hip circumference, and body mass index) were taken and a battery of questionnaires that assessed barrier, task and scheduling self-efficacy, HRQL, and stage of change administered at baseline, week 5 (end of intervention for group A), week 9 (end of intervention for group B; follow-up for group A) and week 13 (follow-up for both groups). Results showed that this workplace physical activity intervention was successful at increasing the participants' daily steps, that task self-efficacy is a significant predictor of participants' exercise adherence during the initial stages of participation (intervention phase), and that the participants felt that this intervention was effective. Finally, further exploratory analyses showed that this intervention was effective for all participants, but most valuable for participants most in need of improvement - that is, those who were most sedentary prior to the intervention. This intervention is an inexpensive use of simple and effective tools (e.g. pedometers), has the potential to attract a wide variety of participants and become a pennanent part of any health promotion initiative.
Resumo:
An in vitro investigation of some important factors controlling the activity of chitin synthase in cell-free extracts of two Mortierella species has been carried out. Mixed membrane fractions from mycelial homogenates of Mortierella candelabrum and Mortierella pusilla were found to catalyse the transfer of N-acetylglucosamine from UDP-N-acetylglucosamine into an insoluble product characterized as chitin by its insolubility in weak acid and alkali, and the release of glucosamine and diacetylchitobiose on hydrolysis with a strong acid and chitinase, respectively. Apparent Km values for UDP-GlcNAc were 1.8 mM and 2.0 mM for M. pusilla and ~ candelabrum, respectively. Polyoxin D was found to be a very potent competitive inhibitor with values of the constant of inhibition, Ki' for both species about three orders of magnitude lower than theKm for UDP-GlcNAc. A divalent cation, Mg+2 , Mn+2 or Co+2 , was required for activity. N-acetylglucosamine, the monomer of chitin, stimulated the activity of the enzyme. The crude enzyme preparation of ~ candelabrum, unlike that of ~ pusilla, showed an absolute requirement for both Mg+2 and N-acetylglucosamine. Large differences in response to exogenous proteases were noted in the ratio of active to inactive chitin synthase of the two species. A fifteen fold or greater increase was obtained after treatment with acid protease (from Aspergillussaitoi) as compared to a two- to four-fold activation of the M. pusilla membrane preparation treated similarly. During storage at 4°C over 48 hours, an endogenous activation of chitin synthase of ~ pus ilIa was achieved, comparable to that obtained by exogenous protease treatment. The high speed supernatant of both species inhibited the chitin synthase activity of the mixed membrane fractions. The inhibitor of ~ pus ilIa was effective against the pre-activated enzyme whereas that of M. candelabrum inhibited the activated enzyme. Several possibilities are discussed as to the role of the different factors regulating the enzyme activity. The suggestion is made from the properties of chitin synthase in the two species that in vivo a delicate balance exists between the activation and inactivation of the enzyme which is responsible for the pattern of wall growth of each fungus.
Resumo:
A strain of Drosophila melanogaster (mid america stock culture no. hl16) has been reported to be deficient in aldehyde oxidase activity (Hickey and Singh 1982). This strain was characterized during the course of this study and compared to other mutant strains known to be deficient in aldehyde oxidase activity. During the course of this investigation, the hl16 strain was found to be temperature sensitive in its viability. It was found that the two phenotypes, the enzyme deficiency, and the temperature sensitive lethality were the result of two different mutations, both mapping to the X-chromosome. These two mutations were found to be separable by recombination. The enzyme deficiency was found to map to the same locus as the cinnamon mutation, another mutation which affects aldehyde oxidase production. The developmental profile of aldehyde oxidase in the hl16 strain was compared to the developmental profile in the Canton S wild type strain. The aldehyde oxidase activity in adult hl16 individuals was also compared to that of various other strains. It was also found that the aldehyde oxidase activity was temperature sensitive in the adult flies. The temperature sensitive lethality mutation was mapped to position 1-0.1.
Resumo:
Pancreatic deoxyribonuclease preferentially digests active genes during all phases of the cell cycle including mitosis. Recently, a DNAse I-directed in ~ nick translation technique has been used to demonstrate differences in the DNAse I sensitivity of euchromatic and heterochromatic regions of mitotic chromosomes. This ill ~ technique has been used in this study to ask whether facultative heterochromatin of the inactive X chromosome can be distinguished from the active X chromosome in mouse and human tissues. In addition to this, in ~ nick translation has been used to distinguish constitutive heterochromatin in mouse and human mitotic chromosomes. Based on relative levels of DNAse I sensitivity, the inactive X chromosome could not be distinguished from the active X chromosome in either mouse or human tissues but regions of constitutive heterochromatin could be distinguished by their relative DNAse I insensitivity. The use of !D situ nick translation was also applied to tissue sections of 7.5 day mouse embryos to ask whether differing levels of DNAse I sensitivity could be detected between different tissue types. Differences in DNAse I sensitivities were detected in three tissues examined; embryonic ectoderm, an embryo-derived tissue, and two extraembryonic tissues, extraembryonic ectoderm and ectoplacental cone. Embryonic ectoderm and extraembryonic ectoderm nuclei possessed comparable levels of DNAse I sensitivity while ectoplacental cone was significantly less DNAse I sensitive. This suggests that tissue-specific mechanisms such as chromatin structure may be involved in the regulation of gene activity in certain tissue types. This may also shed some light on possible tissue specific mechanisms regulating X chromosome activity in the developing mouse embryo.
Resumo:
Pyruvate dehydrogenase (PDH) plays an important role in regulating carbohydrate oxidation in skeletal muscle. PD H is deactivated by a set of PD H kinases (PD K 1-4) with PDK2 and 4 being the predominant isoforms in skeletal muscle. PDK2 is highly sensitive to pyruvate inhibition, and is the most abundant isoform, while PDKI and 4 protein content are normally lower. This study examined the PDK isoform content and PDHa activation in muscle at rest and 10 and 40 Hz stimulation from PDK2 knockout (PDK2KO) mice to delineate the role of PDK2 in activating the PDH complex during low and moderate intensity muscle contraction. PDHa activity was lower in PDK2KO mice during contraction while total PDK actitvity was -4 fold lower. PDK4 protein was not different, however PDKI partially compensated for the lack of PDK2 and was -56% higher than WT. PDKI is a very potent inhibitor of the PDH complex due to its phosphorylation site specificity and allosteric regulation. These results suggest that the site specificity and allosteric regulatory properties of the individual PDK isoforms are more important than total PDK activity in determining transformation of the complex and PDHa activity during acute muscle contraction.