Characterization of Novel Alkaline Protease producing Bacillus subtilis C3a-FIIRO with Potential for Industrial Application

F.A Fashola,O.I Ibidapo,P.N Nwagala,F.A Orji,A.K Lawal,C.P Augustine,I.C James,O.T Fadipe,S.O Olatope,F.B Aderinwale

doi:10.4314/njb.v38i2.6

Abstract

Microbial alkaline protease is one of the dominant industrial enzymes which function in splitting polypeptides chain of protein into monomers of amino acids and peptides. This study aimed to identify alkaline protease produced by Bacillus sp. Soil samples were aseptically collected from dump sites in FIIRO, Lagos state, Nigeria. The samples were serially diluted, and bacteria were isolated using pour plate method. The resulting isolates were screened and morphologically characterized. The isolate with the highest protease production potential was subjected to biochemical characterization using Analytical Profile Index (API) identification kit system and 16S rRNA sequencing. The selected isolate was used to produce alkaline protease by solid state fermentation using rice bran as a substrate. Out of the 18 bacteria isolated, 11 isolates showed alkaline protease production potential. Isolate C3a-FIIRO was selected for its maximal alkaline protease produced as indicated by a 56 mm zone of clearance. Morphological and biochemical characterization revealed isolate C3a-FIIRO as a member of the genus Bacillus. The 16S rRNA gene sequencing confirmed the isolate as Bacillus subtilis C3a-FIIRO (MW577298) with closest homology to Bacillus subtilis Y17B. The enzyme activity of 6848.171 U/ml ± 0.11 and protein concentration of 152.13 mg/ml ± 0.003 showed that Bacillus subtilis C3a-FIIRO has potential for sustainable alkaline protease production.

Highlights

The dependency of the nation on importation of industrial enzymes implies there is no industry currently producing these enzymes in Nigeria, which positions the nation continuously on the radar of other importing nations with strong prospect for these enzymes
Microbial alkaline protease is one of the dominant industrial enzymes which function in splitting polypeptides chain of protein into monomers of amino acids and peptides
This study aimed to identify alkaline protease produced by Bacillus sp

Summary

Introduction

The dependency of the nation on importation of industrial enzymes implies there is no industry currently producing these enzymes in Nigeria, which positions the nation continuously on the radar of other importing nations with strong prospect for these enzymes. Based on pH, they are classified as acidic, alkaline, and neutral proteases (Ellaiah et al, 2002; Sayaniya & Patel, 2021) Among these three proteases, alkaline protease (EC 3.4.21), which are active at alkaline pH, occupy a pivotal position due to their wide industrial application in food, brewing, dairy, and pharmaceutical industries (Kumar et al, 1999; Varia et al, 2019; Manavalan et al, 2020). Bacillus species have been identified as important industrial microorganisms due to their rapid growth rate, extracellular secretion of enzyme and generally recognized as safe status making them bacterial workhorses in industrial microbial cultivations to produce a variety of enzymes (Pandey et al, 2000; Gouda et al, 2006; Vijayalakshmi et al, 2012; Deb et al, 2013; Karray et al, 2021). The benefit of exploiting microorganisms for industrial applications and the need to encourage biodiversity characterization makes the identification of these organisms important. This study focuses on identifying a novel bacterial strain isolated from the soil with hyper alkaline protease producing potential

Material and Methods

Results and Discussions

Conclusion