Production and characterization of thermostable alkaline protease of Bacillus subtilis (ATCC 6633) from optimized solid‐state fermentation

Abstract

Proteases are the most important group of enzymes utilized commercially in various arenas of industries, such as food, detergent, leather, dairy, pharmaceutical, diagnostics, and waste management, accounting for nearly 20% of the world enzyme market. Microorganisms of specially Bacillus genera serve as a vast repository of diverse set of industrially important enzymes and utilized for the large-scale enzyme production using a fermentation technology. Approximately 30%-40% of the cost of industrial enzymes originates from the cost of the growth medium. This study is attempted to produce protease from Bacillus subtilis (ATCC 6633) after optimization of various process parameters with the aid of solid-state fermentation using a cheap nutrient source such as wheat bran. B. subtilis (ATCC 6633) produces proteases of molecular weight 36 and 20 kDa, respectively, in the fermented medium as evident from SDS zymogram. Alkaline protease activity has been detected with optimum temperature at 50 °C and is insensitive to ethylenediaminetetraacetic acid. This thermostable alkaline protease exhibits dual pH optimum at 7 and 10 with moderate pH stability at alkaline pH range. It preserves its activity in the presence of detergent such as SDS, Tween 20, and Triton X-100 and may be considered as an effective additive to detergent formulation with some industrial importance.

Enzyme production by solid‐state fermentation on soybean meal: A comparative study of conventional and ultrasound‐assisted extraction methods

“…However, recently, numerous studies are inclined to a solid‐state fermentation as a more promising technology. This technique offers numerous advantages, but the most important feature of enzyme production in this mode is the possibility of the utilization of inexpensive substrates, which mostly arose as agro‐industrial by‐products or wastes . These substrates have low commercial value and even impose attention and care regarding their environmental impact.…”

“…Various Bacillus spp. have been found to successfully grow on diverse agricultural wastes and by‐products such as various cereal brans, straws, husks, fruit/ vegetable peels from the food processing industry, and so on .…”

Simultaneous Production of Β-glucanase and Protease From Bacillus Velezensis Strain Identified in the Manure of Piglets.

“…To the best of our knowledge, this is the rst report that Bacillus velezensis has been isolated from the manure of piglets. There were many reports about β-glucanase and protease produced by B. subtilis (17,18). B. velezensis had 99% similarity with B. subtilis, therefore, we speculated that B. velezensis can also produceβ-glucanase and protease.…”

Solid‐state fermentation of Moringa oleifera leaf meal using Bacillus pumilusCICC 10440

“…Commercial cellulases are produced mainly by fungi; however, bacteria have also been considered as robust and versatile enzyme producers due to their high growth rate, stability under extreme conditions, and multienzyme complexes . Bacillus strains are often used in modern biological fermentation technology, as they are non‐toxic and biologically safe . From a commercial standpoint, Bacillus pumilus , Bacillus sp ., Bacillus subtilis subsp.…”

“…15,16 Bacillus strains are often used in modern biological fermentation technology, as they are non-toxic and biologically safe. 17,18 From a commercial standpoint, Bacillus pumilus, Bacillus sp., Bacillus subtilis subsp. and Trichoderma reesei have been found to be potent extracellular enzyme producers; as a result, they were chosen for the fermentation of Moringa oleifera leaf meal in the present study.…”