Optimized production and properties of thermostable alkaline protease from Bacillus subtilis SHS-04 grown on groundnut (Arachis hypogaea) meal

Abstract

Production of alkaline protease from Bacillus subtilis SHS-04 was investigated under different fermentation conditions involving low-cost substrates with the aim of optimizing yield of enzyme. Maximum enzyme production (1616.21 U/mL) was achieved using groundnut meal (0.75%) as nitrogen source and 0.5% glucose as carbon source at 48 h cultivation period, pH 9, 45˚C and 200 rpm. The yield was 348% increase over comparable control samples. The alkaline protease had optimum temperature of 60˚C and remarkably exhibited 80% relative activity at 70˚C. It was highly thermostable showing 98.7% residual activity at 60˚C after 60 minutes of incubation at pH 9.0 and was stable in the presence of organic solvents studied. These properties indicate the viability of the protease for biotechnological and industrial applications. The optimized yield of enzyme achieved in this study establishes groundnut meal as potential low-cost substrate for alkaline protease production by B. subtilis SHS-04.

Degradation of chlorothalonil through a hydrolytic dehalogenase secreted from Bacillus subtilis WB800

“…Among these reports, Olajuyigbe obtained the maximum production of alkaline protease from B. subtilis SHS-04, showing a 3.48-fold increase over control samples after optimization (Olajuyigbe, 2013). B. subtilis is a strain with native feature of protease production; thus, higher activity would be achieved more easily for proteases.…”

Enhanced production of alkaline protease from novel bacterium Bacillus cereus GVK21 under submerged fermentation

“…7) examined, groundnut oil cake enhanced the protease production from B. cereus GVK21 by 112 % (1762 U/ml) as compared to control (665 U/ml), i.e., groundnut oil cake supported maximum protease production. Bacillus subtilis SHS-04exhibited maximum protease production (1616.21 U/mL) by utilizing groundnut cake as substrate (Olajuyigbe, 2013).…”

“…Groundnut oil cake, a rich protein source (30-40%), which mainly constitutes different amino acids such as arginine (11.0), Leucine (6.1), glycine (6.0), and phenyl alanine (4.9) and a dry matter of 92.6% comprehends slow release of nitrogen due to its conditioned and moderately complex nature which may favor the process organism metabolically and physiologically for effi cient production of protease (Kuo, 1967). Several reports indicate that groundnut cake serves as a reasonably good nitrogen source for protease production by various microorganisms (Kuberan et al, 2010;Kranthi et al, 2013, Olajuyigbe, 2013.…”

“…Utilization of agro industrial residues as carbon and nitrogen sources for the bulk production of industrial enzymes may play a signifi cant role not only in reducing the production cost and also contribute towards growing environmental concerns by addressing the agro industrial waste disposal and man-agement problems because of tremendous quantities of agricultural residues generated through agricultural practices and industrial processes (Bajaj and Wani, 2011;Singh and Bajaj 2015). Earlier workers also reported that defatted oil meals like soybean meal (Saurabh et al, 2007), cotton seed cake (Bajaj et al, 2013) and groundnut meals (Olajuyigbe, 2013) are the cost-effective alternatives for industrial production processes. Hence utilization of a low-priced nitrogen source is an important criterion for economic production of industrial enzymes.…”

Optimization of fermenting medium by statistical method for production of alkaline protease by Bacillus licheniformis MZK05M9

“…To produce alkaline protease, many researchers used inexpensive substrates coupled with expensive one as a carbon and nitrogen source such as glucose and soybean meal [15][16][17], soybean meal and trypton [18], wheat bran and soybean meal [19], starch and soybean meal [20], molasses and potassium nitrate [21], molasses and urea [22], starch, wheat bran and soya flour [23], wheat bran and beef extract [24], and glucose and groundnut meal [25]. However, there are fewer attempts reported to induce protease production using inexpensive both carbon and nitrogen sources [2,26].…”