34 resultados para Enzimas - Regulação
Resumo:
Nowadays generation ethanol second, that t is obtained from fermentation of sugars of hydrolyses of cellulose, is gaining attention worldwide as a viable alternative to petroleum mainly for being a renewable resource. The increase of first generation ethanol production i.e. that obtained from sugar-cane molasses could lead to a reduction of lands sustainable for crops and food production. However, second generation ethanol needs technologic pathway for reduce the bottlenecks as production of enzymes to hydrolysis the cellulose to glucose i.e. the cellulases as well as the development of efficient biomass pretreatment and of low-cost. In this work Trichoderma reesei ATCC 2768 was cultivated under submerged fermentation to produce cellulases using as substrates waste of lignocellulosic material such as cashew apple bagasse as well as coconut bagasse with and without pretreatment. For pretreatment the bagasses were treated with 1 M NaOH and by explosion at high pressure. Enzyme production was carried out in shaker (temperature of 27ºC, 150 rpm and initial medium pH of 4.8). Results showed that T.reesei ATCC 2768 showed the higher cellulase production when the cashew apple bagasse was treated with 1M NaOH (2.160 UI/mL of CMCase and 0.215 UI/mL of FPase), in which the conversion of cellulose, in terms of total reducing sugars, was of 98.38%, when compared to pretreatment by explosion at high pressure (0.853 UI/mL of CMCase and 0.172 UI/mL of Fpase) showing a conversion of 47.39% of total reducing sugars. Cellulase production is lower for the medium containing coconut bagasse treated with 1M NaOH (0.480 UI/mL of CMcase and 0.073 UI/mL of FPase), giving a conversion of 49.5% in terms of total reducing sugars. Cashew apple bagasse without pretreatment showed cellulase activities lower (0.535 UI/mL of CMCase and 0,152 UI/mL of FPase) then pretreated bagasse while the coconut bagasse without pretreatment did not show any enzymatic activity. Maximum cell concentration was obtained using cashew nut bagasse as well as coconut shell bagasse treated with 1M NaOH, with 2.92 g/L and 1.97 g/L, respectively. These were higher than for the experiments in which the substrates were treated by explosion at high pressure, 1.93 g/L and 1.17 g/L. Cashew apple is a potential inducer for cellulolytic enzymes synthysis showing better results than coconut bagasse. Pretreatment improves the process for the cellulolytic enzyme production
Resumo:
The production of enzymes by microorganisms using organic residues is important and it can be associated with several applications such as food and chemical industries and so on. The objective of this work is the production of CMCase, Xylanase, Avicelase and FPase enzymes by solid state fermentation (SSF) using as substrates: bagasse of coconut and dried cashew stem. The microorganisms employed are Penicillium chrysogenum and an isolated fungus from the coconut bark (Aspergillus fumigatus). Through the factorial design methodology and response surface analysis it was possible to study the influence of the humidity and pH. For Penicillium chrysogenum and the isolated fungus, the coconut bagasse was used as culture medium. In another fermentation, it was used the mixture of coconut bagasse and cashew stem. Fermentations were conducted using only the coconut bagasse as substrate in cultures with Penicillium chrysogenum fungus and the isolated one. A mixture with 50% of coconut and 50% of cashew stem was employed only for Penicillium chrysogenum fungus, the cultivation conditions were: 120 hours at 30 °C in BOD, changing humidity and pH values. In order to check the influence of the variables: humidity and pH, a 2 2 factorial experimental design was developed, and then two factors with two levels for each factor and three repetitions at the central point. The levels of the independent variables used in ascending order (-1, 0, +1), to humidity, 66%, 70.5% and 75% and pH 3, 5 and 7, respectively. The software STATISTICA TM (version 7.0, StatSoft, Inc.) was used to calculate the main effects of the variables and their interactions. The response surface methodology was used to optimize the conditions of the SSF. A chemical and a physic-chemical characterization of the coconut bagasse have determined the composition of cellulose (%) = 39.09; Hemicellulose (%) = 23.80, Total Lignin (%) = 36.22 and Pectin (%) = 1.64. To the characterization of cashew stem, the values were cellulose (g) = 15.91 Hemicellulose (%) = 16.77, Total Lignin (%) = 30.04 and Pectin (%) = 15.24. The results indicate the potential of the materials as substrate for semisolid fermentation enzyme production. The two microorganisms used are presented as good producers of cellulases. The results showed the potential of the fungus in the production of CMCase enzyme, with a maximum of 0.282 UI/mL and the Avicelase enzyme the maximum value ranged from 0.018 to 0.020 UI/ mL, using only coconut bagasse as substrate. The Penicillium chrysogenum fungus has showed the best results for CMCase = 0.294 UI/mL, FPase = 0.058 UI/mL, Avicelase = 0.010 UI/mL and Xylanase = 0.644 UI/ mL enzyme, using coconut bagasse and cashew stem as substrates. The Penicllium chrysogenum fungus showed enzymatic activities using only the coconut as substrate for CMCase = 0.233 UI/mL, FPase = 0.031 to 0.032 UI/ mL, Avicelase = 0.018 to 0.020 UI/mL and Xylanase = 0.735 UI/ mL. Thus, it can be concluded that the used organisms and substrates have offered potential for enzyme production processes in a semi-solid cultivation
Resumo:
The need for new sources of energy and the concern about the environment have pushed the search for renewable energy sources such as ethanol. The use of lignocellulosic biomass as substrate appears as an important alternative because of the abundance of this raw material and for it does not compete with food production. However, the process still meets difficulties of implementation, including the cost for production of enzymes that degrade cellulose to fermentable sugars. The aim of this study was to evaluate the behavior of the species of cactus pear Opuntia ficus indica and Nopalea cochenillifera, commonly found in northeastern Brazil, as raw materials for the production of: 1) cellulosic ethanol by simultaneous saccharification and fermentation (SSF) process, using two different strains of Saccharomyces cerevisiae (PE-2 and LNF CA-11), and 2) cellulolytic enzymes by semi-solid state fermentation (SSSF) using the filamentous fungus Penicillium chrysogenum. Before alcoholic fermentation process, the material was conditioned and pretreated by three different strategies: alkaline hydrogen peroxide, alkaline using NaOH and acid using H2SO4 followed by alkaline delignification with NaOH. Analysis of composition, crystallinity and enzymatic digestibility were carried out with the material before and after pretreatment. In addition, scanning electron microscopy images were used to compare qualitatively the material and observe the effects of pretreatments. An experimental design 2² with triplicate at the central point was used to evaluate the influence of temperature (30, 40 and 45 °C) and the initial charge of substrate (3, 4 and 5% cellulose) in the SSF process using the material obtained through the best condition and testing both strains of S. cerevisiae, one of them flocculent (LNF CA-11). For cellulase production, the filamentous fungus P. chrysogenum was tested with N. cochenillifera in the raw condition (without pretreatment) and pretrated hydrothermically, varying the pH of the fermentative medium (3, 5 and 7). The characterization of cactus pear resulted in 31.55% cellulose, 17.12% hemicellulose and 10.25% lignin for N. cochenillifera and 34.86% cellulose, 19.97% hemicellulose and 15.72% lignin for O. ficus indica. It has also been determined, to N. cochenillifera and O. ficus indica, the content of pectin (5.44% and 5.55% of calcium pectate, respectively), extractives (26.90% and 9.69%, respectively) and ashes (5.40% and 5.95%). Pretreatment using alkaline hydrogen peroxide resulted in the best cellulose recovery results (86.16% for N. cochenillifera and 93.59% for O. ficus indica) and delignification (48.79% and 23.84% for N. cochenillifera and O. ficus indica, respectively). This pretreatment was also the only one which did not increase the crystallinity index of the samples, in the case of O. ficus indica. However, when analyzing the enzymatic digestibility of cellulose, alkali pretreatment was the one which showed the best yields and therefore it was chosen for the tests in SSF. The experiments showed higher yield of conversion of cellulose to ethanol by PE-2 strain using the pretreated N. cochenillifera (93.81%) at 40 °C using 4% initial charge of cellulose. N. cochenillifera gave better yields than O. ficus indica and PE-2 strain showed better performance than CA-11. N. cochenillifera proved to be a substrate that can be used in the SSSF for enzymes production, reaching values of 1.00 U/g of CMCase and 0.85 FPU/g. The pretreatment was not effective to increase the enzymatic activity values
Resumo:
Shrimp farming in Brazil is a consolidated activity, having brought economical and social gains to several states with the largest production concentrated in the northeast. This fact is also reflected in higher feed intake, necessitating a more efficient feed management. Currently, management techniques already foresee food loss due to molting. In this sense, studies relating shrimp s digestive physiology, molting physiology and behavioral response of shrimp feed can optimize the feed management. Thus, our study aimed to evaluate the behavioral response of the marine shrimp L. vannamei (Crustacea: Penaeidae) in accordance with the stages of moulting cycle and feeding schedules based on higher or lower activity of proteolytic digestive enzymes; also, to investigate the influence of feeding schedule on hepatosomatic index and non-specific and specific protease activity (trypsin). Experiments were carried out at the Laboratory of Shrimp Behavioral Studies at UFRN in partnership with the Laboratory of Enzimology UFPE. Juveniles of L. vannamei weighting 5.25 g (+ 0.25 g) were kept in aquaria at a density of 33 shrimp m -2. In the first experiment, shrimp were fed in the light phase or in the dark phase for 8 days; in the ninth day, the animals were observed for 15 minutes every hour during the 12 hours of each phase of the photoperiod. We recorded the frequency of inactivity, exploration, food intake, burrowing, swimming and crawling behavior. At the end of the 12th observation session, the shrimp were sacrified and classified by the method of setogenesis in the molt cycle stages A, B, C, D0, D1, D2 or D3. We found that the shrimp in A stage show high levels of inactivity. Moreover, the frequency of food intake was very low. The shrimp in D3 stage also had low food intake and high inactivity associated with elevated frequencies of burrowing. In the second experiment, shrimp were kept in physiological acclimation to experimental conditions for 28 days, distributed in 12 treatments in the light phase and 12 treatments in the dark phase. In the end, the animals were sacrified and dissected to assess non-specific and specific protease activity (trypsin) activity. In general, these parameters did not vary among animals fed in the light phase and those fed in the dark phase. However, significant differences were found in the activity of specific and nonspecific proteases in relation to food treatment. In the light phase, the major proteolytic activities converged to 10 hours after the start of the light phase, while the lowest activities converged to 6 hours after the beginning of this phase. In the dark phase, the highest enzyme activity converged to 12 hours after the onset of phase, while the lowest activities converged to 3 hours after the onset of phase. In the third experiment, we sought to evaluate the behavioral responses of shrimp in relation to dietary treatments based on higher or lower activity of proteolytic enzymes, considering the results of the second experiment. The behavioral categories observed were the same as the ones in the first experiment, with observations of 30 minutes (15min before and 15min after food supply). We found variation in behavioral responses as a function of the treatments, with greater intake of food in shrimp fed during the period of greatest activity of proteolytic enzymes, in the light phase. Thus we see that periodic events associated with the shrimp s physiology interfere in their behavioral responses, revealing situations that are more adjustable to the provision of food, and consequently optimizing feeding management
Resumo:
The concern with issues related to consumer protection has emerged in North America and then spread throughout the world. In Brazil, consumer‟s rights and interests only gained greater importance after their consolidation in the Constitution of 1988 and the enactment of the 8078/90 Law (Consumer‟s Protection and Defense Code), which established the consumerist microsystem. The understanding of the legal relationship of consumption concept is necessarily connected to knowledge of the elements that compose it. Among these, we can find the consumer and the provider (subjective elements), the product or service (objective elements), and the consumer‟s condition as final receiver of the consumption object (finalistic element). In order to elucidate the configuration of consumer protection before advertising communication, this work will analyze the advertising through the prism of consumerist laws, conceptualizing it and presenting a differentiation of it in relation to practices such as marketing, offer and commercial communication as well as examining its several kinds of manifestation, focusing mainly the ones categorized as misleading or unfair advertising. All kinds of advertising communication against the consumerist microsystem are subject to judicial control exercised by the State. Besides individual protection possibilities, this state-owned control can be collectively exercised as a result of the utilization of public civil action and popular action. Some specific categories of advertising (smoking products, alcoholic beverages, pesticides, medicines and therapies) are still subject to a set of particular restraints provided by the 9294/96 Law, which enables the performance of a special control in relation to them. In addition to state control, there is also a system of advertising communication self-regulation, which develops itself through the actions of the National Council of Advertising Self-Regulation that are based mainly on the laws established by the Brazilian Code of Advertising Self-Regulation and its annexes. However, this system of advertising self-regulation still has some deficiencies that hinder its effectiveness
Resumo:
The present work aims to demonstrate the link of the principle of efficiency - as expressed in the Constitution of 1988, by Constitutional Amendment No. 19 - with regulatory agencies, more specifically the ANATEL (National Telecommunications Agency). It also includes this principle’s importance to regulation - to monitor and manage public services - as well as when an activity will be considered efficient, keeping in mind that agencies are subjected to other principles of public administration. The increasing use of telephony has enabled further development of technologies that provide improvements in the provision of this service. The VoIP (Voice over IP), is nothing more than a technological breakthrough that directly targets the providers of conventional telephone service, both by modifying the business working for a long time with the same technology as the amount of new competitors’ dispute on market share. It also analyses the difficulty of understanding and definition of what is VoIP telephony, its growth and the threats that the traditional and mostly which is ANATEL’s role concerning this telephony technology. As regulator of the telecommunications service, ANATEL not yet regulated the voice telephony service using the IP protocol. What looks over the years is that ANATEL exercise its regulatory function to provide better conditions for competition among providers of VoIP and traditional telephone companies, obviously some difficulties are expected, given that VoIP is a technology that provides two services, through conventional telephony and using the internet.
Resumo:
While essential to human nature, health and life have been protected since ancient times by various areas of knowledge, particularly by the Law, given its dynamics within the regulation of social interactions. In Brazil, health has been granted major importance by the Federal Constitution of 1988, which, disrupting the dictatorial authoritarianism, inaugurating a Social State and focusing on the values of freedom and human dignity, raises health to the condition of a social right, marked predominantly by an obligational bias directed, primarily, to the State, through the enforcement of public policies. Although, given the limitation of the State action to the reserve for contingencies, it turns clear that an universalizing access to public health is impossible, seen that the high cost of medical provisions hinders the State to meet all the health needs of the rightholders. As a result of the inefficiency of the State, the effort of the Constituent Assembly of 1988 in creating a hybrid health system becomes nuclear, which, marked by the possibility of exploration of healthcare by the private initiative, assigns to the private enterprise a key role in supplementing the public health system, especially through the offer of health insurance plans. At this point, however, it becomes clear that health provisions rendered by the private agents are not unlimited, which involves discussions about services and procedures that should be excluded from the contractual coverage, for purposes of sectoral balance, situation which draws the indispensability of deliberations between Fundamental Rights on one hand, related to the protection of health and life, and contractual principles on the other hand, connected to the primacy of private autonomy. At this point, the importance of the regulation undertaken by the ANS, Brazilian National Health Agency, appears primordial, which, by means of its seized broad functions, considerable autonomy and technical discretion, has conditions to implement an effective control towards the harmonization of the regulatory triangle, the stability and development of the supplementary health system and, consequently, towards the universalization of the right to health, within constitutional contours. According to this, the present essay, resorting to a broad legislative, doctrinal and jurisprudential study, concludes that economic regulation over the private healthcare sector, when legitimately undertaken, provides progress and stability to the intervening segment and, besides, turns healthcare universalization feasible, in a way that it can not be replaced efficiently by any other State function.
Resumo:
In Brazil, the National Agency of Electric Energy (ANEEL) represents the energy regulator. The rates review have been one of its main tasks, which establish a pricing practice at a level to cover the efficient operating costs and also the appropriate return of the distributors investments. The changes in the procedures to redefine the efficient costs and the several studies on the methodologies employed to regulate this segment denote the challenge faced by regulators about the best methodological strategy to be employed. In this context, this research aims to propose a benchmarking evaluation applied to the national regulation system in the establishment of efficient operating costs of electricity distribution utilities. The model is formulated to promote the electricity market development, partnering with government policies ant to society benefit. To conduct this research, an integration of Data Envelopment Analysis (DEA) with the Stochastic Frontier Analysis (SFA) is adopted in a three stages procedure to correct the efficiency in terms of environmental effects: (i) evaluation by means of DEA to measure operating costs slacks of the utilities, in which environmental variables are omitted; (ii) The slacks calculated in the first stage are regressed on a set of environmental variables by means of SFA and operating costs are adjusted to account the environmental impact and statistical noise effects; and, (iii) reassess the performance of the electric power distribution utilities by means of DEA. Based on this methodology it is possible to obtain a performance evaluation exclusively expressed in terms of management efficiency, in which the operating environment and statistical noise effects are controlled.
Resumo:
Objective Based on the system of reference and counter-reference and comprehensiveness in oral health care, we aimed to examine ways of refering users to Specialized Dental Care Centers (SDCC) and the interface between them and Primary Care. Methods This is a cross-sectional study carried out with users and dentists of SDCC in a metropolitan region of Northeast of Brazil. Analyses were descriptive, and the association test was done with chi-square. Results Six forms of entry to specialized service were identified: free demand (13.8 %) and reference by the Primary Care dentist (63.2 %) were most frequent. Users referred by the basic health unit dentist had more interest in making a counter-reference than the others (p<0.001, PR=4.65, 95 % CI: 2.74 to 7.91), while individuals without this referral had 1.49 times more difficulty obtaining care (95 % CI: 1.02 to 2.17). Referral procedures are a decisive factor for counter-references. However, the high demand for primary care services and the short supply these services can offer in the face of needs make SDCC performance difficult. Conclusion The analysis of oral health practices from the perspective of network modeling points to the service's need to establish protocols for regulation in a bid to improve access to and the quality of care provided.
Resumo:
Objective Based on the system of reference and counter-reference and comprehensiveness in oral health care, we aimed to examine ways of refering users to Specialized Dental Care Centers (SDCC) and the interface between them and Primary Care. Methods This is a cross-sectional study carried out with users and dentists of SDCC in a metropolitan region of Northeast of Brazil. Analyses were descriptive, and the association test was done with chi-square. Results Six forms of entry to specialized service were identified: free demand (13.8 %) and reference by the Primary Care dentist (63.2 %) were most frequent. Users referred by the basic health unit dentist had more interest in making a counter-reference than the others (p<0.001, PR=4.65, 95 % CI: 2.74 to 7.91), while individuals without this referral had 1.49 times more difficulty obtaining care (95 % CI: 1.02 to 2.17). Referral procedures are a decisive factor for counter-references. However, the high demand for primary care services and the short supply these services can offer in the face of needs make SDCC performance difficult. Conclusion The analysis of oral health practices from the perspective of network modeling points to the service's need to establish protocols for regulation in a bid to improve access to and the quality of care provided.
Resumo:
The global energy crisis triggered the possibility of unconventional hydrocarbons exploration and production, culminating in the US energy revolution as well as making other countries interested in the development of these natural resources. The justification for this research comes from Brazil's interest in the use of hydraulic fracturing techniques in unconventional resources since the Brazilian National Agency of Petroleum, Natural Gas and Biofuels (ANP) organized the 12th acquisition process and published the Resolution no. 21 of April 10, 2014. The conflict between economic viability and the social and environmental damage from exploration and production of unconventional hydrocarbons in Brazil resulted in the search for a legal solution that would consider the economic, social and environmental interests. The main purpose here is to analyze the regulation of unconventional oil exploration and production in Brazil in order to show lack of regulatory instruments so far. The specific objectives are to investigate how the lack of effective regulation may ultimately prevent the development itself, analyze the importance of systematization of a new regulatory tool for ensuring legal security and energy, identify the key negative environmental and social impacts, and suggest possibilities approaches within the new regulatory framework. The research methodology stands out the hypothetico-deductive model as approach, and the comparative model as procedural method. Moreover, the research techniques used here are performance of a theoretical and descriptive questioning over literature search, analysis of Brazilian standardization and case laws, and a brief comparative study, in order to provide suggested approaches for a new regulatory framework.
Resumo:
The global energy crisis triggered the possibility of unconventional hydrocarbons exploration and production, culminating in the US energy revolution as well as making other countries interested in the development of these natural resources. The justification for this research comes from Brazil's interest in the use of hydraulic fracturing techniques in unconventional resources since the Brazilian National Agency of Petroleum, Natural Gas and Biofuels (ANP) organized the 12th acquisition process and published the Resolution no. 21 of April 10, 2014. The conflict between economic viability and the social and environmental damage from exploration and production of unconventional hydrocarbons in Brazil resulted in the search for a legal solution that would consider the economic, social and environmental interests. The main purpose here is to analyze the regulation of unconventional oil exploration and production in Brazil in order to show lack of regulatory instruments so far. The specific objectives are to investigate how the lack of effective regulation may ultimately prevent the development itself, analyze the importance of systematization of a new regulatory tool for ensuring legal security and energy, identify the key negative environmental and social impacts, and suggest possibilities approaches within the new regulatory framework. The research methodology stands out the hypothetico-deductive model as approach, and the comparative model as procedural method. Moreover, the research techniques used here are performance of a theoretical and descriptive questioning over literature search, analysis of Brazilian standardization and case laws, and a brief comparative study, in order to provide suggested approaches for a new regulatory framework.
Resumo:
A Constituição Federal de 1988 e o Estatuto da Cidade estabelecem o Plano Diretor como instrumento básico da política de ordenamento territorial, tendo como princípio fundamental o cumprimento da função social da propriedade e do direito à cidade. Na perspectiva de adequação às diretrizes e objetivos da política urbana estabelecidos em 1988, o município de Natal elaborou os Planos Diretores de 1994 e 2007, definindo instrumentos e parâmetros de regulação do uso e ocupação do solo possíveis de assegurar o cumprimento da função social da propriedade urbana e de gerar subsídios ao planejamento e à gestão da cidade. Apesar de Natal ter sido um dos municípios brasileiros pioneiros na adoção desses princ pios, antecipando e incorporando os instrumentos que em 2001 viriam a ser definidos no Estatuto da Cidade, identifica-se que alguns desses instrumentos e parâmetros direcionados à regulação do uso e ocupação do solo não tiveram sua aplicação plena, a exemplo do mecanismo de acompanhamento e controle dado pelo Estoque de Área Edificável e da Densidade, que foi substituída pelo Coeficiente de Aproveitamento no Plano Diretor de 2007. Questionando esse procedimento, busca-se na presente pesquisa investigar de que maneira essa substituição do parâmetro densidade pelo coeficiente de aproveitamento influenciou na capacidade da gestão pública de regular os processos de uso e ocupação do solo, de forma a adequar a sua intensificação ao suporte da infraestrutura instalada. Foram tomadas como referência teórico-conceitual as contribuições sobre a prática de planejamento urbano no Brasil, nos marcos do ideário da reforma urbana, com destaque para as reflexõe s de Flávio Villaça, Orlando Alves Santos Junior e Daniel Todtmann Montandon, Luiz César de Q. Ribeiro, Raquel Rolnik, Ermínia Maricato, Laura Machado de Bueno e Renato Cymbalista, José Roberto Bassul e Carlos F. Lago Burnett, e, com relação aos parâmetros de controle urbanístico, o estudo identifica as diferentes abordagens sobre a densidade urbana e o coeficiente de aproveitamento com base nas reflexões de Claudio Acioly Jr., Forbes Davidson, Juan Luis Mascaró, Ricardo Ojima, Marcelo de Souza, José Rámon Navarro Vera e Armando Ortuño Padilla, Nestor Goulart Reis, Marta Dora Grostein e Susana Ricardo Alves. Como conclusão, discute-se a hipótese formulada, inicialmente, de que a mudança de parâmetros verificada colocou limites para o município realizar uma gestão adequada do solo urbano e, portanto, de fazer cumprir a função social da propriedade, considerando a necessidade de adequação entre a intensificação do uso e ocupação do solo e a infraestrutura instalada
Resumo:
Serines proteinases inhibitors (PIs) are widely distributed in nature and are able to inhibit both in vitro and in vivo enzymatic activites. Seed PIs in than leguminous are classified in seven families, Bowman-Birk and Kunitz type families that most studied representing an important role in the first line of defense toward insects pests. Some Kunitz type inhibitors possess activities serine and cysteine for proteinases named bifunctional inhibitor, as ApTKI the inhibitor isolate from seed of Adenanthera pavonina. The A. pavonina inhibitor presenting the uncommon property and was used for interaction studies between proteinases serine (trypsin) and cysteine (papain). In order to determinate the in vitro interaction of ApTKI against enzymes inhibitor purification was carried cut by using chromatographic techniques and inhibition assays. The 3D model of the bifunctional inhibitor ApTKI was constructed SWISS-MODEL program by homology modeling using soybean trypsin inhibitor (STI, pdb:1ba7), as template which presented 40% of identity to A. pavonina inhibitor. Model quality was evaluated by PROCHECK program. Moreover in silico analyzes of formed complex between the enzymes and ApTKI was evaluated by HEX 4.5 program. In vitro results confirmed the inhibitory assays, where the inhibitor presented the ability to simultaneously inhibit trypsin and papain. The residues encountered in the inhibitor model of folder structural three-dimensional that make contact to enzymes target coud explain the specificity pattern against serine and cysteine proteinases
Resumo:
A β-D-N-acetilglucosaminidase extracted and partially isolated from crustacean Artemia franciscana by ammonium sulfate precipitation and filtration gel chromatography Bio Gel A 1.5m. the enzyme was immobilized on ferromagnetic Dacron yielding a insoluble active derivative with 5.0 units/mg protein and 10.35% of the soluble enzyme activity. β-D-N-acetilglucosaminidase-ferromagnetic Dacron was easily removed from the reaction mixture by a magnetic field, it was reused for ten times without loss in its activity. The ferromagnetic Dacron was better activated at pH 5.0. The particles visualized at scanning electron microscope (SEM) had presented different sizes, varying between 721nm and 100µm. Infra red confirmed immobilization on support, as showed by primary amino peaks at 1640 and 1560 cm-1 . The immobilize enzyme presented Km of 2.32 ± 0.48 mM and optimum temperature of 50°C. Bought presented the same thermal stable of the soluble enzyme and larger enzymatic activity at pH 5.5. β-D-N-acetilglucosaminidase-Dacron ferromagnético showed sensible for some íons as the silver (AgNO3), with loss of activity. The β-D-N acetilglucosaminidase activity for mercury chloride (HgCl2), whom is one of the most toxic substance joined in nature, it was presented activity already diminished at 0,01mM and lost total activity at 4mM, indicating sensitivity for this type of metal. β-D-N-acetilglucosaminidase-ferromagnetic Dacron showed degradative capacity on heparan sulfate, the enzyme still demonstrated degradative capacity on heparan sulphate, suggesting a possible application to produce fractions of this glycosaminoglycan