Abstract

Proteases are important industrial enzymes that account for about 60% of the total enzyme market globally due to their large application in food, feed, textile and pharmaceutical industries. The effect of salt stress on protease production was evaluated on Aspergillus flavus and Aspergillus niger. The enzyme production was enhanced by stepwise optimization of the culture parameters, notably, carbon source, nitrogen source, pH, and temperature of the submerged fermentation process while using a minimal salt media and casein as substrate for the protease activity. The fungi species were found to be good producers of both acid and alkaline proteases under 4% salt stress condition. The optimum culture conditions for alkaline protease production by Aspergillus flavus were sucrose 4%, peptone 1%, pH 8 at 40°C with maximum enzymatic activities of 8.85 mM/min/mg protein, 5.22 mM/min/mg protein, 3.75 mM/min/mg protein, and 1.64 mM/min/mg protein, respectively. Lactose 4%, peptone 1%, pH 6 at 50°C were the optimum culture conditions for acid protease production by Aspergillus flavus with maximum enzymatic activities of 4.59 mM/min/mg protein, 2.06 mM/min/mg protein, 1.24 mM/min/mg protein, and 1.23 mM/min/mg protein, respectively. For Aspergillus niger, the optimum culture conditions for alkaline protease production were corn starch 4%, yeast extract 1%, pH 6 at 40°C with maximum enzymatic activities of 5.99 mM/min/mg protein, 3.85 mM/min/mg protein, 6.18 mM/min/mg protein, and 3.72 mM/min/mg protein, respectively. While lactose 4%, yeast extract 1%, pH 6 at 50°C were the best culture conditions for acid protease production by Aspergillus niger with maximum enzymatic activities of 4.81 mM/min/mg protein, 0.93 mM/min/mg protein, 5.71 mM/min/mg protein, and 3.34 mM/min/mg protein, respectively.

Highlights

  • Microbial proteases are degradative enzymes, which catalyse the total hydrolysis of proteins [1]

  • Corn starch with an enzymatic activity of 5.99 mM/min/mg protein served as the best carbon source for extracellular alkaline protease production from Aspergillus niger (Figure 1)

  • During the preliminary stage of this study, the NaCl concentration in the minimal salt media was varied at an interval of 2% from 0% to 6% for alkaline and acid protease production by Aspergillus flavus and Aspergillus niger (Figure S1)

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Summary

Introduction

Microbial proteases are degradative enzymes, which catalyse the total hydrolysis of proteins [1]. Proteases are the most important industrial enzyme accounting for about 60% of the total enzyme market in the world and approximately 40% of the total worldwide enzyme sale [2]. They are very essential in the production of detergents for protein stain removal [3] [4]. Proteases are essential for improving the functional, nutritional and flavour properties in proteins especially in baking where they are used to degrade proteins in flour for biscuits, crackers and cookies. Proteases give a wide application such as in treatment of clotting disorder [5] and in the production of digestive and certain medical treatments of inflammation and virulent wounds [6]

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