Optimization of alkaline protease production from Bacillus subtilis NS isolated from sea water

Abstract

The protease producing bacterial strain Bacillus subtilis was isolated from sea water and identified by 16S rRNA sequencing. The strain named as B. subtilis NS. Optimization of the strain revealed that the most suitable nitrogen source to enhance protease production was beef extract. Among various carbon sources tested, maximum production of protease was registered in medium with added glucose. The effect of metals ions indicated that maximum protease production was observed in medium supplemented with magnesium chloride (MgCl). Investigating the effect of sodium chloride (NaCl) concentration on protease production revealed that 7% yielded higher protease production. The most suitable pH and temperature for maximum protease production revealed that pH 9 and a temperature of 40°C gave optimal protease production.

Microbial Proteases Applications

“…These enzymes have gained interest not only due to their vital role in metabolic activities but also due to their immense utilization in industries (Rao et al, 1998; Sandhya et al, 2005; Younes and Rinaudo, 2015). The proteases available in the market are of microbial origin because of their high yield, less time consumption, less space requirement, lofty genetic manipulation, and cost-effectiveness, which have made them suitable for biotechnological application in the market (Nisha and Divakaran, 2014; Ali et al, 2016). These microbial proteases are preferred to plant and animal proteases because of the presence of all desired characteristics for industrial applications (Palsaniya et al, 2012; Sathishkumar et al, 2015).…”

The fermentation optimization for alkaline protease production by Bacillus subtilis BS-QR-052

“…Notably, corn syrup powder, as a composite nitrogen source, emerged as a significant factor influencing the yield, closely linked to the secretion of extracellular protease. Additionally, the inclusion of inorganic salts and metal ions contributed to the maintenance of extracellular alkaline protease stability, aligning with the pronounced impact observed with the addition of MgSO 4 (Nisha and Divakaran, 2014). Hence, the search for an optimal combination of cultivation medium components holds great promise in promoting the maximum secretion of extracellular alkaline protease by B. subtilis.…”

“…High productivity is one of the advantages of B. subtilis in producing alkaline protease. The enzyme activity of alkaline protease produced by wild strains ranges from 50 to 500 U/mL ( Nisha and Divakaran, 2014 ; Bukhari et al, 2020 ), and can be increased to 500–1,000 U/mL through optimization of the culture medium or overexpression methods ( Shafique et al, 2021 ; Emon et al, 2023 ). In this study, the enzyme activity was 598 U/mL and 1780.03 U/mL before and after optimization, respectively, representing a remarkable 198% improvement.…”

“…Nitrogen sources commonly used are soybean meal, peptone, urea, yeast extract, beef paste, soybean cake powder, and corn syrup powder. Previous research suggests that a combination of carbon and nitrogen sources may be more suitable for enhancing the fermentation process and maximizing alkaline protease production by B. subtilis ( Krishnaveni et al, 2012 ; Nisha and Divakaran, 2014 ; Sathishkumar et al, 2015 ). In this study, we employed a composite carbon source (corn starch) and a composite nitrogen source (corn syrup powder) to optimize the production yield of alkaline protease.…”

Screening and isolation of protease-producing bacteria from wastewater samples in Obafemi Awolowo University (OAU) Campus, Ile-Ife, Nigeria

“…These enzymes have gained interest not only due to their vital role in metabolic activities but also due to their immense utilization in industries (Rao et al, 1998;Sandhya et al, 2005;Younes & Rinaudo, 2015). The proteases available in the market are of microbial origin because of their high yield, less time consumption, less space requirement, lofty genetic manipulation, and cost-effectiveness, which have made them suitable for biotechnological application in the market (Nisha & Divakaran, 2014;Ali et al, 2016). Comparatively, proteases produced by plants and animals are more labor-intensive than microbially produced proteases (Gupta et al, 2002;Kalaiarasi & Sunitha, 2009).…”