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The present study evaluated the influence of the variables polyethylene glycol (PEG) molar mass, pH, PEG concentration and sodium citrate concentration in the integrated production of the protease ...

β-fructofuranosidases (FFases) are enzymes involved in sucrose hydrolysis and fructo-oligosaccharides' production which are of great interest for the food industry. FFase from Aspergillus tamarii URM4634 was extracted using PEG/Phosphate Aqueous Biphasic Systems (ABS), and the impact of magnetic field on the extraction behavior was evaluated. A 24-full experimental design was employed to study the influence of molar mass of PEG, concentrations of PEG and phosphate and pH on the selected response variables, i.e., partition coefficient (K), purification factor (PF), activity yield (Y) and selectivity (S). The influence of magnetic field during partition and NaCl concentration on the same responses was also studied. The best results of FFase extraction without magnetic field (K = 0.50, PF = 4.05, Y = 72.66% and S = 0.06) were observed at pH 8.0 using 12.5% (w/w) PEG 400 and 25% (w/w) NaH2PO4/K2HPO4. Application of the magnetic field allowed improving the performance, with the best results being obtained at the longest distance between magnets (lowest magnetic field) and absence of NaCl (K = 0.93, PF = 4.22, Y = 83.79% and S = 0.09). The outcomes obtained demonstrate that ABS combination with low intensity magnetic field can be used as an efficient FFase pre-purification method.

A Caatinga e um bioma exclusivamente brasileiro, com fauna e flora peculiares, proporcionando assim um ambiente propicio para micro-organismos adaptaveis a nutricao e clima adversos. O processo de obtencao de enzimas atraves dos fungos por meio de fermentacao em estado solido e bastante viavel por manter o micro-organismo em condicoes que simulam o ambiente natural. Assim sendo, o presente trabalho teve como objetivo realizar a bioprospeccao de enzimas produzidas por Aspergillus tamarii URM4634 isolado do solo da caatinga. O micro-organismo foi mantido com o meio de cultura Czapek por 7 dias a 30°C ate ocorrer esporulacao. Os esporos foram coletados, padronizados na concentracao de 107 esporos/mL e inoculados no substrato de farelo de trigo (FT). A fermentacao em estado solido foi realizada contendo 5g de substrato FT, previamente autoclavado a 121°C por 20 minutos, com um teor de 40% de umidade, por 72 horas, a 30°C em câmara climatica. Apos a fermentacao foi extraido o extrato bruto (EB), adicionando Apos a fermentacao o extrato proteolitico foi obtido por adicao de 7,5 mL de fosfato de sodio 0,1M, pH 7,0 por grama de material fermentado e homogeneizado na mesa agitadora por 2 horas, submetido maceracao durante 1 minuto, agitacao orbital 5000 rpm durante 15 minutos a 4°C e em seguida filtrado/centrifugado. O EB foi analisado para diferentes atividades enzimaticas: protease, colagenase, endoglucanase, pectinase e frutosiltransferase. Nas determinacoes das atividades enzimaticas realizadas, verificou-se altos valores para protease com 116,6 U/mL, podemos destacar a producao de colagenase com 210,6 U/mL. Ainda pode ser verificada atividade de 0,41 (U/mL) de endoglucanase; 0,49 (U/mL) pectinases e 10,7 (U/mL) de frutosiltransferase. O perfil obtido apresentou nove bandas proteicas com pesos moleculares de 14,4 a 97,0 kDa. A natureza do composto proteolitico (enzima) foi confirmada por zimograma para as atividades de frutosiltransferase e protease, onde a banda hidrolisada branca correspondeu a posicao da frutosiltransferase, sendo visualizado duas bandas com pesos moleculares distintos, indicando assim a presenca de duas diferentes enzimas com a mesma atividade. Para a atividade proteolitica foi observado a presenca de apenas uma banda. Os resultados demostraram que A. tamarii URM4634, foi capaz de produzir todas as enzimas testadas, com substrato de baixo custo utilizando subprodutos agroindustriais. Tendo um maior potencial de utilizacao industrial para as proteases, no qual foi obtido altos valores de atividade, em destaque para a producao de colagenases.

β-fructofuranosidases (FFases) are enzymes involved in sucrose hydrolysis and fructo-oligosaccharides' production which are of great interest for the food industry. FFase from

The present study evaluated the influence of the variables polyethylene glycol (PEG) molar mass, pH, PEG concentration and sodium citrate concentration in the integrated production of the protease from

β-fructofuranosidases (EC3.2.1.26) are members of the GH32 family of glycoside hydrolases, which include more than 390 enzymes of vegetable and microbial origins, used in several biotechnological applications. Thus, this research aimed to produce a β-fructofuranosidase obtained by Aspergillus tamarii through solid state fermentation, and to purify by Aqueous Two-Phase System (ATPS). Summary results presented the optimal parameters to produce the β-fructofuranosidase used wheat bran as a substrate at 30 °C for 48 h, and purification process using ATPS with polyethylene glycol and sodium citrate (PEG/sodium citrate), where the β-fructofuranosidase preferably partitioned to the salt-rich phase, the best run (24% of PEG 400, 20% sodium citrate, pH 8) which presented a higher purification factor 6.42 with 12.39 U/mL activity and 352% yield. Optimum parameter was pH 5.15 and temperature of 55 °C, respectively. The purified enzyme showed excellent thermal stability and exhibited a half-life of 60 min at 65 °C. Kinetics results for enzyme showed for Sucrose substrate the enzyme showed Km of 42.9 ± 2.21 mM and Vmax of 180.2 ± 2.8 μM min−1 mg−1 of protein. Although Vmax was the highest for 1-Kestose (219.4 ± 2.7 μM min−1 mg−1 of protein) the preferred substrate of Aspergillus tamarii β-fructofuranosidase (β-FFase) was Nystose (Km of 3.8 ± 0.15 mM). SDS-PAGE revealed a single band of protein at ~66 kDa. Finally, this study demonstrated the potential of ATPS to purify a β-fructofuranosidase with application in biotechnological field aiming to functional foods.

RESUMO As proteases fibrinolíticas são capazes de degradar coágulos de fibrina formados dentro dos vasos sanguíneos, evitando a trombose intravascular. Em animais, a tromboflebite, que acomete frequentemente os equinos, ocasiona, em seus casos graves, a obstrução jugular e também um edema de laringe, derivando a obstrução das vias aéreas, o que possibilita um edema cerebral, ocorrendo o óbito do animal. Devido ao fato de o tratamento ser de custo elevado, faz-se necessária a investigação de outras fontesde proteases fibrinolíticas com custos menores e com menos efeitos colaterais. Diante disso, este estudo tem como objetivo produzir e caracterizar proteases fibrinolíticas obtidas de Streptomyces parvulus DPUA 1573. Para produção da enzima, foi utilizado um planejamento fatorial 24 avaliando a concentração da farinha de soja (0,5, 1,0 e 1,5%) e da glicose (0, 0,5 e 1,0g/L), temperatura (28, 32 e 37ºC) e agitação (150, 200 e 250rpm) sobre a biomassa e a atividade fibrinolítica. Pode-se verificar que a protease fibrinolítica apresentou atividade máxima (835U/mL) nas condições de concentração de 1,5% de soja, 1g/L de glicose, 28°C e 150rpm com 48 horas de fermentação. A protease fibrinolítica obtida teve temperatura e pH ótimos de 55°C e pH 9,0, respectivamente. A atividade enzimática foi inibida pelo EDTA, pelo íon Fe2+ e pelo SDS, o que indicou a enzima ser uma metaloprotease. A linhagem Streptomyces parvulus DPUA 1573 foi capaz de produzir protease fibrinolítica, possuindo características bioquímicas favoráveis à aplicação na medicina veterinária e possivelmente humana. ABSTRACT Fibrinolytic proteases are able to degrade fibrin clot formed in the blood vessel, avoiding intravascular thrombosis. In animals, thrombophlebitis often affects horses, and in severe cases causes obstruction of the jugular and laryngeal edema leading to airway obstruction allowing cerebral edema resulting in the death of the animal. Since treatment is costly, the investigation of other sources of fibrinolytic proteases at lower cost and with fewer side effects is needed. Thus, this study aims to produce and characterize fibrinolytic proteases from Streptomyces parvulus DPUA 1573. For enzyme production, a factorial design was performed to evaluate 24 soybean flour concentration (0.5, 1.0 and 1.5%) and glucose (0, 0.5 and 1.0g/L), temperature (28, 32 and 37°C) and agitation (150, 200 and 250rpm) on biomass and fibrinolytic activity. Fibrinolytic protease showed maximum activity (835 U/mL) under these conditions: 1.5% soybean flour, 1g/L glucose, 28°C, and 150rpm 48 hours of fermentation. The optimal temperature was 55°C and optimal pH was 9.0. Fibrinolytic protease activity was inhibited by EDTA, the ion Fe2+, and by SDS, which indicated that the enzyme is a metallo-protease. The strain Streptomyces parvulus DPUA 1573 was able to produce fibrinolytic protease with biochemical characteristics favorable for application in veterinary and human medicine.

Resumo: A mastite é uma inflamação na glândula mamária que pode acarretar perdas na produção e na qualidade do leite, gerando prejuízos econômicos para a pecuária leiteira. O tratamento é baseado na utilização de antibióticos, sendo, em muitos casos, ineficazes devido à resistência bacteriana já conhecida para esta doença. O objetivo deste trabalho foi selecionar linhagens de Streptomyces spp. produtoras de biocompostos com atividade antimicrobiana frente a isolados do gênero Staphylococcus multirresistentes de búfalas com mastite. Bem como, determinar os melhores parâmetros de produção, e avaliar a produção simultânea de ácido clavulânico. A seleção de Streptomyces spp. com capacidade de produzir compostos com atividade antimicrobiana foi realizada através da técnica bloco de gelose. Dentre as 30 espécies de Streptomyces spp. testadas, o micro-organismo Streptomyces parvulus DPUA 1573 apresentou melhores resultados, sendo capaz de inibir o crescimento de 7 isolados Staphylococcus spp. multirresistentes. Posteriormente, a espécie selecionada Streptomyces parvulus DPUA 1573 foi cultivada em diferentes condições pré-determinadas pelo planejamento fatorial 24, onde as variáveis independentes foram: concentração de soja (0,5; 1,0; 1,5%), glicose (0; 0,5; 1g/L), agitação (150; 200; 250rpm) e temperatura (28; 32; 37°C) e todos os ensaios do planejamento foram monitorados até 120 horas de cultivo. Todas as variáveis independentes influenciaram positivamente no crescimento celular, enquanto que para atividade antimicrobiana apenas as variáveis temperatura e agitação apresentaram efeitos significativos positivos. O líquido metabólito produzido por Streptomyces parvulus DPUA 1573 foi capaz de inibir o crescimento de sete Staphylococcus spp. multirresistentes. As melhores condições de cultivo para a produção de moléculas bioativas por este micro-organismo foi a 37?C, com 250rpm de agitação por período de 72 horas. Nos ensaios que apresentaram atividade antimicrobiana, foi avaliada a produção de ácido clavulânico ao longo do cultivo. A maior concentração de ácido clavulânico foi de 269,84g/L obtidas nas condições de 1,5% de farinha de soja em ausência de glicose no tempo de 96 horas. A linhagem Streptomyces parvulus DPUA 1573 foi eficiente contra Staphylococcus spp. multirresistentes isolados de mastite em búfalas, ainda apresentando concomitantemente produção de ácido clavulânico com o potencial uso farmacêutico. Abstract: Mastitis is an inflammation in one or more mammary glands which can lead to reduction in production and quality of milk causing economic losses to dairy farming. The use of antibiotics is the key for the treatment of this disease, but in many cases ineffective due to bacterial resistance already known for this condition. The aim of this study was to select strains of Streptomyces spp. producing biomolecules with antimicrobial activity against multidrug-resistant Staphylococcus isolated from buffaloes with mastitis, as well as to determine the best production parameters to the evaluation of simultaneous production of clavulanic acid. Thirty species of Streptomyces spp. were used to selecting the greatest producer spectrum of antimicrobial activity (agar block technique), with selection of Streptomyces parvulus DPUA 1573, and 7 multidrug-resistant Staphylococcus spp. sensitive to its biocompounds. The selected strain of Streptomyces parvulus DPUA 1573 was cultured in different conditions predetermined by the factorial design 24, where the independent variables were: soybean concentration (0.5, 1.0, 1.5%), glucose (0, 0.5, 1g/L), agitation (150, 200, 250rpm) and temperature (28, 32, 37°C); all the tests were monitored up to 120 hours of cultivation. All independent variables influenced positively the cell growth, while for antimicrobial activity only the variables temperature and agitation showed positive effects. The antimicrobial bio compounds showed activity against seven multidrug-resistant Staphylococcus spp under the conditions: temperature 37°C, agitation 250rpm, with 72 hours of production process. In the tests which showed antimicrobial activity, was also assessed the production of clavulanic acid along with the cultivation. The highest concentration of clavulanic acid was 269.84g/L obtained under the conditions of 1.5% of soybean flour and absence of glucose in 96 hours. The strain Streptomyces parvulus DPUA 1573 was effective against multidrug-resistant strains of Staphylococcus spp. of mastitis from buffaloes, still showing concomitantly production of clavulanic acid for pharmaceutical use.

A fibrinolytic protease from M. subtilissimus UCP 1262 was recovered and partially purified by polyethylene glycol (PEG)/sodium sulfate aqueous two-phase systems (ATPS). The simultaneous influence of PEG molar mass, PEG concentration and sulfate concentration on the enzyme recovery was first investigated using a 23 full factorial design, and the Response Surface Methodology used to identify the optimum conditions for enzyme extraction by ATPS. Once the best PEG molar mass for the process had been selected (6000 g/mol), a two-factor central composite rotary design was applied to better evaluate the effects of the other two independent variables. The fibrinolytic enzyme was shown to preferentially partition to the bottom phase with a partition coefficient (K) ranging from 0.2 to 0.7. The best results in terms of enzyme purification were obtained with the system formed by 30.0% (w/w) PEG 6000 g/mol and 13.2% (w/w) sodium sulfate, which ensured a purification factor of 10.0, K of 0.2 and activity yield of 102.0%. SDSPAGE and fibrin zymography showed that the purified protease has a molecular mass of 97 kDa and an apparent isoelectric point of 5.4. When submitted to assays with different substrates and inhibitors, it showed selectivity for succinyl-l-ala-ala-pro-l-phenylalanine-p-nitroanilide and was almost completely inhibited by phenylmethylsulfonyl fluoride, behaving as a chymotrypsin-like protease. At the optimum temperature of 37° C, the enzyme residual activity was 94 and 68% of the initial one after 120 and 150 min of incubation, respectively. This study demonstrated that M. subtilissimus protease has potent fibrinolytic activity compared with similar enzymes produced by solid-state fermentation, therefore it may be used as an agent for the prevention and therapy of thrombosis. Furthermore, it appears to have the advantages of low cost production and simple purification., The authors acknowledge the financial support of the Brazilian Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES), Fundacao de Amparo a Ciencia e Tecnologia do Estado de Pernambuco (FACEPE), and Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq). The authors also thank the project approved in the REN-NORFUN network (MCT/CNPq/MMA/MEC/CAPES/FNDCT, Acao Transversal/FAPs, No.47/2010, Sistema Nacional de Pesquisa em Biodiversidade - SISBIOTA/Brazil).

Fibrinolytic enzymes have received attention regarding their medicinal potential for thrombolytic diseases, a leading cause of morbidity and mortality worldwide. Various natural enzymes purified from animal, plant and microbial sources have been extensively studied. The aim of this work was to produce fibrinolytic protease by solid state fermentation using agro industrial substrates. Rhizopus arrhizus var. arrhizus UCP 1295 and Mucor subtillissimus UCP 1262 filamentous fungi species isolated from soil of Caatinga-PE, Brasil, were used as producer microorganisms. Wheat bran was shown to be the best substrate for the production of the enzyme and by using a 23 full factorial design the main effects and interactions of the quantity of the substrate wheat bran, moisture and temperature on the fibrinolytic enzyme production and protease were evaluated. The best results for fibrinolytic and protease activities, 144.58 U/mL and 48.33 U/mL, respectively, were obtained with Mucor subtillissimus UCP 1262 using as culture medium 3 g wheat bran, 50% moisture at a temperature of 25°C for 72 hours. The optimum temperature for the produced enzyme was 45°C and most of its original activity was retained after being subjected to 80°C for 120 min. The protease activity was enhanced by K+, Ca+ and Mn+; but with Cu+ there was an inhibition. The specificity to chromogenic substrate and the inhibition by PMSF indicates that it is a chymotrypsin-like serine protease. Presented results suggest that this enzyme produced by solid-state fermentation is an interesting alternative as a candidate for thrombolytic therapy.

Parkia pendula seed lectin was used to treat cutaneous wounds of normal and immunocompromised mice, inducing cicatrization. Methotrexate (0.8 mg/kg/week) was used as immunosuppressive drug. Wounds were produced in the dorsal region (1 cm(2)) of female albino Swiss mice (Mus musculus), health and immunocompromised. Wounds were daily topically treated with 100 μL of the following solutions: (1) control (NaCl 0.15 M), (2) control Im (0.15 M NaCl), (3) P. pendula seed lectin (100 μg/mL), and (4) P. pendula seed lectin Im (100 μg/mL). Clinical evaluation was performed during 12 days. Biopsies for histopathology analysis and microbiological examinations were carried out in the second, seventh, and 12th days. The presence of edema and hyperemia was observed in all groups during inflammatory period. The first crust was detected from the second day, only in the groups treated with P. pendula seed lectin. Microbiological analysis of wounds from day 0 to day 2 did not show bacterium at P. pendula seed lectin group; however, Staphylococcus sp. was detected every day in the other groups. The lectin markedly induced a total wound closing at P. pendula seed lectin and P. pendula seed lectin Im groups on 11th day of evolution. The present study suggests that P. pendula seed lectin is a biomaterial potential to show pharmacological effect in the repair process of cutaneous wounds.

This work explores the possibility of using a polyethylene glycol (PEG)/sodium citrate aqueous two-phase system (ATPS) as a first step in a process for the purification of lectin from Canavalia grandiflora seed. Purification of lectins is a limiting step for its uses in the area of biotechnology. Extraction by an aqueous two-phase system is a powerful technique for separation, concentration, and purification of biomolecules and pharmaceutical products. Four factors (PEG's molar mass, PEG's concentration, pH, citrate concentration) affecting the lectin ConGF partitioning were studied. A two-level factorial design (24) was carried out. Lectin ConGF preferentially partitioned to the top (polyethylene glycol) phase. The statistical analysis showed that the citrate concentration chloride significantly affects the KL (partitioning coefficient for protein) value for lectin partitioning. An ATPS composed of 20% (w/w) PEG 400 and 20% (w/w) citrate, at pH 6, allowed the recovery of lectin ConGF with an 8.67 partition coefficient and 104% yield. Consequently, the system proved to be efficient and can be used as a first step in lectin purification from crude extract of ConGF seeds.

The ascorbate oxidase is the enzyme used to determine the content of ascorbic acid in the pharmaceutical and food industries and clinics analyses. The techniques currently used for the purification of this enzyme raise its production cost. Thus, the development of alternative processes and with the potential to reduce costs is interesting. The application of aqueous two-phase system is proposed as an alternative to purification because it enables good separation of biomolecules. The objective of this study was to determine the conditions to continuously pre-purify the enzyme ascorbate oxidase by an aqueous two-phase system (PEG/citrate) using rotating column provided with perforated discs. Under the best conditions (20,000 g/mol PEG molar mass, 10% PEG concentration, and 25% citrate concentration), the system showed satisfactory results (partition coefficient, 3.35; separation efficiency, 54.98%; and purification factor, 1.46) and proved suitable for the pre-purification of ascorbate oxidase in continuous process.

This work deals with the use of an aqueous two-phase system (ATPS) of PEG/citrate to remove proteases from a Clostridium perfringens fermentation broth. To plan the experimental tests and evaluate the corresponding results, three successive experimental designs were employed, for which the PEG molar mass (MPEG) and concentration (CPEG), the citrate concentration (CC) and the pH were selected as independent variables, while the purification factor (PF), the partition coefficient (K), the activity yield (Y) and the selectivity (S) were selected as responses. PF of proteases in the top phase was shown to increase with increasing MPEG and decreasing CC, whereas a completely opposite trend was observed for K. On the other hand, Y was favored by simultaneous decreases in both these variables, while S decreased with increasing CC. Therefore, selecting a simultaneous increase in PF and Y as the most desirable result, the best performance of the system was obtained using MPEG = 10,000 g/mol, CPEG = 22% (w/w) and CC = 8.0% (w/w) at pH 8.5. Under these conditions, the activity yield was very high (131%) but the purification factor (4.2) and the selectivity (4.3) were lower than those ensured by more selective purification methods. According to these results, the ATPS seems to be an interesting alternative primary concentration/decontamination step for vaccine preparation from C. perfringens fermented broth.

Polysaccharide obtained from Anacardium occidentale L. gum was used for trypsin entrapment using cellulose (gaze) as a support and this preparation was applied as cutaneous wound healing. Trypsin release in vitro and the influence of pH and temperature on activity, stability and storage time of entrapped enzyme were evaluated. The preparation showed that it was still capable to release enzyme even after 48 h. Entrapped enzyme presented an optimal pH and temperature of 8.6 and 55 °C, respectively. Also, it was stable at high temperature (45 °C for 60 min) and wide range of pH, retaining 80% of its initial activity when stored for 28 days at 25 °C. Histopathological analysis of mice skin wound healing under the entrapped trypsin preparation treatment showed an acceleration of fibroblast proliferation, neovascularization of granulation tissue and stimulating effect on the epithelium formation compared to the skin wound under the treatment using preparations without trypsin. These results demonstrate that the trypsin–polysaccharide–cellulose preparation could be used in cutaneous dressing applications for wound healing.

A novel protease with fibrinolytic activity was purified from MucorsubtilissimusUCP 1262usinga two-step purification protocol.The enzyme was pre-purified using cetonic precipitation and adsorbed by ion exchange chromatography on DEAE-sephadex G50. System two-dimensional electrophoresis (2DE) coupled to SDS-PAGE showed a single protein band of15.3kDa approximatelyand isoeletric focusing point of 3.9, exhibiting a nature as an acidic enzyme.The activity was suppressed by Co 2+ and HgCl2, but slightly enhanced byCa 2+ . Additionally, the activity was slightly inhibited by EDTA, but significantlyinhibited by PMSF. It exhibited fibrinolytic activity,which is weaker than that of plasmin, but also had a higher affinity for the N-Succinyl-Ala-Ala-Pro-Phe p-nitroanilide (SAAPNA) and azocasein substrates, suggesting a chymotrypsin-like protease. These results demonstrated an economically viable protocol purification of a protease with fibrinolytic activity with high degree of purity that may represent a potential source of new therapeutic agents to treat thrombosis.

The kinetic and thermodynamic properties of ascorbate oxidase (AO) activity and stability of a Cucurbita maxima extract were investigated. Activity tests performed at 25 degrees C using initial ascorbic acid concentration in the range 50-750 M allowed estimating the Michaelis constant for this substrate (Km = 126 microM) and the maximum initial rate of ascorbic acid oxidation (A0,max = 1.57 mM min-1). The main thermodynamic parameters of the enzyme reaction (DeltaH* = 10.3 kJ mol-1; DeltaG* = 87.2 kJ mol-1; DeltaS* = -258 J mol-1 K-1) were estimated through activity tests performed at 25-48 C. Within such a temperature range, no decrease in the initial reaction rate was detected. The long-term thermostability of the raw extract was then investigated by means of residual activity tests carried out at 10-70 degrees C, which allowed estimating the thermodynamic parameters of the irreversible enzyme inactivation as well (DeltaH*D = 51.7 kJ mol-1; DeltaG*D = 103 kJ mol-1; S*D = -160 J mol-1 K-1). Taking into account the specific rate of AO inactivation determined at different temperatures, we also estimated the enzyme half-life (1047 min at 10 degrees C and 21.2 min at 70 degrees C) and predicted the integral activity of a continuous system using this enzyme preparation. This work should be considered as a preliminary attempt to characterize the AO activity of a C. maxima extract before its concentration by liquid-liquid extraction techniques.

Cramoll 1,4 is a lectin extracted from seeds of Cratylia mollis Mart. Many assays have shown the cytokine release activity and pro-inflammatory profile of this lectin. Here, we used Cramoll 1,4 in the treatment of cutaneous wounds in normal and immunocompromised mice for available your cicatricial power. Surgical wounds were treated daily with a topical administration of Cramoll 1,4 and parameters as edema, hyperemia, scab, granulation and scar tissues as well as contraction of wounds were analyzed. Cramoll 1,4 wounds showed higher edema and arrival of more polimorphonuclear cells at the site of lesions. Granulation tissue and collagen fiber deposition were observed with higher intensity in all Cramoll 1,4 treated wounds and promoted excellent closing and repair of lesions in less time than other groups. Results showed that Cramoll 1,4 lectin was effective in the repair of experimental lesions in mice and can be used as a future cicatricial compound.