&lt;b&gt;Production and partial characterization of proteases from &lt;i&gt;Mucor hiemalis&lt;/i&gt; URM3773

Roana Cecília Dos Santos Ribeiro,Erika Valente Medeiros,Thaís Rafaelle Dos Santos Ribeiro,Keila Aparecida Moreira,Cristina Maria De Souza-Motta

doi:10.4025/actascibiolsci.v37i1.21016

Abstract

The current study evaluated the proteases production from 11 fungal species belonging to the genera Mucor, Rhizomucor and Absidia. The species were obtained from the Collection of Cultures URM at the Mycology Department-UFPE, Brazil. The best producing species was Mucor hiemalis URM 3773 (1.689 U mL -1 ). Plackett-Burman design methodology was employed to select the most effective parameter for protease production out of 11 medium components, including: concentration of filtrate soybean, glucose, incubation period, yeast extract, tryptone, pH, aeration, rotation, NH4Cl, MgSO4 and K2HPO4. Filtrated soybean concentration was the significant variable over the response variable, which was the specific protease activity. The crude enzyme extract showed optimal activity in pH 7.5 and at 50oC. The enzyme was stable within a wide pH range from 5.8 to 8.0, in the phosphate buffer 0.1M and in stable temperature variation of 40-70oC, for 180 minutes. The ions FeSO4, NaCl, MnCl2, MgCl2 and KCl stimulated the protease activity, whereas ZnCl2 ion inhibited the activity in 2.27%. Iodoacetic acid at 1mM was the proteases inhibitor that presented greater action.The results indicate that the studied enzyme have great potential for industrial application.

Highlights

The world has witnessed a significant increase in the use of enzymes as industrial catalysts
In particular, represent an important group of industrially produced enzymes and they account for 60% of the worldwide sales value of the total industrial enzymes
The thermal stability of the evaluated enzyme extract was higher when compared with other reported proteases, such as: Hirsutella rhossiliensis, in which the enzyme was stable for 2 hours between 40 and 50°C (WANG et al, 2009) and Monacrosporium microscaphoides that were incubated at 70°C for 10 minutes and resulted in a reduction of 90% in the proteolytic activity (MIAO et al, 2006)

Summary

Introduction

The world has witnessed a significant increase in the use of enzymes as industrial catalysts. The biggest sector in the biotechnological industry relies on the production and use of microbial origin enzymes (BON et al, 2008). It is essential to optimize the fermentation medium for proteases growth and production, in order to obtain high and commercially viable yields (ESPÓSITO et al, 2009). It is essential to know the enzyme properties, such as its stability, optimal working conditions and mechanistic class. Such knowledge is essential before considering its potential industrial uses (BALQIS; ROSMA, 2011). The present study aimed to quantitatively select the best filamentous fungus proteases producing species that does it by submerged fermentation in order to obtain crude proteases preparation. Temperature optimum performance, pH and temperature stability as well as the effects of the metal ions and of proteases inhibitors

Material and methods

Results and discussion

Conclusion