ABSTRACT
As enzimas fibrinolíticas podem ser obtidas de micro-organismos por meio de processos fermentativos. O presente trabalho teve como objetivo avaliar a produção e extração integrada da protease fibrinolítica de Mucor subtilissimus UCP 1262 usando sistema de duas fases aquosas (SDFA). O processo integrado foi realizado para avaliar a produção, partição e recuperação da protease fibrinolítica, segundo planejamento experimental 23, utilizando como variáveis independentes a massa molar do polietileno glicol (PEG), a concentração do PEG e a concentração do sulfato de sódio. A maior atividade fibrinolítica (15,40U/mL) foi obtida na fase rica em sulfato de sódio no ensaio composto por 10% de sal e 18% de PEG 8000 (g/mol). Recuperações superiores a 80% foram obtidas. A protease fibrinolítica apresentou pH ótimo 7,0, estabilidade entre os pH 6,0 e 8,5, temperatura ótima 50°C, sendo estável de 10°C a 50°C. A enzima foi classificada como uma serino protease, com massa molecular de 52kDa. Como resultado, o processo é notavelmente eficaz para pré-purificar a protease fibrinolítica com baixo custo e rapidez significativa. Quando comparada a outras técnicas de produção e purificação isoladas, a fermentação extrativa é um processo digno a ser substituto das etapas iniciais de separação convencionais.(AU)
Fibrinolytic enzymes can be obtained from microorganisms through fermentative processes. The study aimed to evaluate the fibrinolytic protease production and integrated extraction from Mucor subtilissimus UCP 1262 by extractive fermentation using Aqueous Two-Phase Systems (ATPS). The integrated process was carried out to assess the production, partition and fibrinolytic enzyme recovery, according to a 2 3 -experimental design, using as independent variables Polyethylene glycol (PEG) molar mass, PEG and sodium sulphate concentration, concentration. The highest fibrinolytic activity (15.40U/mL) was obtained in sodium sulfate rich phase in the assay comprising of 10% of salt and 18% of PEG 8000 (g/mol). Yield greater than 80% was obtained. The fibrinolytic protease presented optimum pH 7.0 and stability between pH 6.0 and 8.5, and optimum temperature 50°C, stable between 10°C to 50°C. The enzyme was classified as a serine-protease with 52kDa of molecular weight. As a result, the process is remarkably effective to pre-purify the fibrinolytic protease with a low cost and significantly faster processing time. When compared to other isolated production and purification techniques the extractive fermentation is worthy of being a candidate to replace the initial stages of conventional separation processes.(AU)
Subject(s)
Fibrin/antagonists & inhibitors , Fibrinolytic Agents/isolation & purification , Mucor/enzymology , Enzyme Induction , FermentationABSTRACT
La regeneración ósea guiada (ROG) es un procedimiento que consiste en el incremento de la cantidad del hueso empleando materiales poliméricos biocompatibles como por ejemplo, el quitosano y el plasma rico en fibrina (PRF); los cuales se han valorado de manera individual con excelentes resultados. En este estudio, se propone analizar radiográficamente la regeneración ósea de ambos polímeros sobre alvéolos dentales postextracción. Se seleccionaron 5 pacientes con indicación de extracción de cordales inferiores bilaterales y a un alvéolo se aplicó quitosano y al otro PRF; se realizaron radiografías periapicales a los 15, 30, 60 y 120 días. Posteriormente, se analizaron las radiografías para observar el nivel de regeneración ósea y los resultados mostraron que ambos biomateriales regeneraron los tejidos, pero con la siguiente diferencia: la ROG con PRF ocurrió en menor tiempo mientras que la ROG con quitosano tuvo una mejor organización estructural. Se concluye que ambos biomateriales se pueden tomar como opciones de tratamientos en la regeneración ósea guiada de tejidos.
The bone regeneration it a procedure that consists in the formation of new bony tissues using biocompatible polymeric materials, among them the chitosan (natural biopolymer) and the rich fibrin plasma (PRF) this biomaterials have been studied and used to achieve the bony regeneration obtaining excellent results. The aim of this research was to design an experiment that allowing us to compare the regenerator effect of both polymers, after the extraction of dental pieces in human beings, using periapical radiography's. Five patients, with indication of extraction of the lower bilateral wisdom dental pieces were selected. Chitosan and PRF were applied on different dental sockets after surgery. To register the information, periapical Rx were taken and analyzed, the procedure were done monthly, for 120 days. The results for Rx showed that both biomaterials regenerated bony tissue. The PRF generated major quantity of tissue in minor time whereas the chitosan did it with better structural organization, so, it was possible to conclude that chitosan and PRF represents interesting options for the treatments where guided bony regeneration is required.