Produção de protease colagenolítica por Bacillus stearothermophillus de solo amazônico

Abstract

RESUMOAs proteases colagenolíticas são enzimas capazes de hidrolisar as ligações peptídicas de vários tipos de colágeno e têm grande importância na medicina e em aplicações terapêuticas. O objetivo desta pesquisa foi avaliar a produção de proteases colagenolíticas por Bacillus stearothermophilus. Os tratamentos foram realizados por meio de um planejamento fatorial completo 2 3 , a fim de avaliar a significância dos efeitos e interações das variáveis -pH inicial, concentração de substrato e temperatura -sobre a produção de protease colagenolítica. O ponto central foi executado em quadruplicata para fornecer uma estimativa dos erros experimentais. Ensaios enzimáticos com colágeno e azocaseína como substratos foram realizados para determinação das atividades colagenolítica e proteolítica respectivamente. A maior atividade enzimática colagenolítica foi 79,38 U mL , em condições iniciais de fermentação, na concentração de substrato a 1% (p/v), pH 7,2 e 25 °C. A atividade proteolítica da enzima foi mais ativa em pH 9,0 e 50 °C e foi estável nas faixas de pH (6,0 -9,0) e temperatura (45 °C -50 °C). Bacillus stearothermophilus apresenta viabilidade para a produção de proteases colagenolíticas e a obtenção dessas enzimas tem grande importância para aplicações biotecnológicas. PALAVRAS-CHAVE: enzimas bacterianas, colágeno, planejamento fatorial Production of collagenolytic protease by Bacillus stearothermophillus from an Amazonian soil ABSTRACT Collagenolytic proteases are enzymes able of hydrolyzing the peptide bonds of several types of collagen, and have great importance in the medicine and therapeutic applications. The aim of this research was to evaluate the production of collagenolytic proteases by Bacillus stearothermophilus. Treatments were performed used a 2 3 full factorial design, in order to evaluate the significance of the effects and interactions of variables -initial pH, substrate concentration and temperature -on the production of collagenolytic protease. The central point was run in quadruplicate to provide an estimate of the experimental error. Enzyme assays with collagen and azocasein as substrates were performed to determine the collagenolytic and proteolytic activities respectively. The highest collagenolytic enzyme activity was 79.38 U mL -1 , corresponding to a specific activity of 136.86 U mg -1 , in the initial fermentation conditions, substrate concentration at 1% (w/v), pH 7.2 and 25 °C. Proteolytic activity of enzyme was most active at pH 9.0 and 50 °C, and was stable over wide pH (6.0 -9.0) and temperature ranges (45 °C -50 °C). Bacillus stearothermophilus shows viability for the production of collagenolytic proteases, and the obtaining these enzymes have high importance for biotechnological applications.

Rapid microbiological tests for prospecting new fungal strains with high potentiality for the pectinolytic enzymes production

“…Microbial enzymes are widely used in industrial processes possessing a wide application in the conversion of food, in pharmaceutical production of medicines, leather industry, detergents, textiles and in the management of industrial waste (Lima, 2014), may be obtained by bacteria, yeasts and mainly by filamentous fungi from genus Aspergillus that are capable of producing enzymes in culture medium of low cost, secreting extracellularly. These fungi present formation of septate and branched mycelium, white or yellowish coloration with formation of stalks (Guimarães et al, 2006;Lelis et al, 2012;Haas et al, 2013).…”

Isolation and purification of Bacillus thuringiensis var. israelensis IMV B-7465 peptidase with specificity toward elastin and collagen

“…Our results are confirmed by the literature data showing that collagenase peptidases exhibit maximum activities at pH 7.0-8.0 in the temperature range from 25 to 65 °C. The highest activity of collagenase of B. stearothermophilus DPUA1729 reported to be at pH 7.2 and 25 °C [14], B. megaterium NRCat pH 7.5 and 50 °C [15], B. licheniformis F11 -at pH 9.0 and 50 °C [16], B. cereus MBL13 -at pH 8.0 and 40 °C [17], B. cereus AT -at pH 8.0 and 50 °C [18]. As it is seen, the majority of bacilli elastases are alkaline enzymes.…”