International Research Journal of Engineering and Technology (IRJET)
e-ISSN: 2395-0056
Volume: 10 Issue: 07 | Jul 2023
p-ISSN: 2395-0072
www.irjet.net
Isolation and identification of protease producing bacteria Divyansh Rana1, Mukesh Kumar Rana2, Sheetal Shirodkar3 1Student, Amity Institute of Biotechnology, Amity University, Noida, Uttar Pradesh, India
2Principal Scientist, Division of Genomic Resources, ICAR-NBPGR, Pusa Campus, New Delhi, India
Amity Institute of Biotechnology, Amity University, Noida, Uttar Pradesh, India ---------------------------------------------------------------------***--------------------------------------------------------------------3Assistant Professor II,
Abstract - Microbes are utilized in numerous industrial
(Kumar et al., 2002). Proteases are present in all organisms including plants, animals, fungi, bacteria, and viruses. Different sources of microbial proteases from which they are isolated and identified mostly include various bacteria and fungi. Most proteases for commercial utilization, are isolated from bacteria belonging to the genus Bacillus. Bacillus megaterium, Bacillus stearothermophilus, Bacillus thuringiensis, Bacillus pumilus, Bacillus subtilis and Bacillus stearothermophilus produce thermostable proteases. Besides Bacillus species, Clostridium histolyticum and various Streptococcus sp., produce cysteine proteases and Vibrio chlorae, Pseudomonas aeruginosa and Streptomyces griseus produce metalloproteases (Gupta et al., 2002; Lakshmi et al., 2008). The proteases obtained from microbial sources hold a pivotal role in industrial applications since it leads to a great reduction in production costs (Sawant and Nagendran, 2014). The production of these protease enzymes in microbes depends on the culture media type and composition used for microbial growth, such as the ratio of carbon to nitrogen, occurrence of a metabolizable sugar such as glucose etc. (Gupta et al., 2002). Besides, protease production depends upon the physiological factors such as the optimum pH, optimum temperature, inoculation media and time of incubation (Suppiah et al., 2012). Even biological characteristics like the genome of an individual can lead to change in the biochemical behavior of the microbe and affect the production of metabolite (Varela et al., 1996). Commercially, optimization of culture media using numerous media constituents during fermentation leads to formation of a cost-effective metabolic yield (Hameed et al., 1999). Also, till date no particular media is established to maximize the production of any enzyme because the genomic diversity present in microbes causes each microbial strain to exhibit a unique environment for maximizing enzymatic production. Hence, it is quite essential to perform a series of experiments for investigating the metabolite or enzyme production pattern under different environmental aspects from new isolated microbial strains to achieve maximum production (Prakasham et al., 2006). The protease producing bacterial identification is the next step after successful isolation and is performed using various morphological and biochemical assays, supplemented by specific tests like antibiotic inhibition and serotyping tests. Recent research led to development of newer molecular diagnostic methodologies which allow microbial strain identification using their genetic sequences, directly from the source sample. Under biochemical characterization, the
applications in a number of ways that exploit their own metabolic capabilities to aid human benefits. Proteases, the enzymatic product of bacteria, act as an integral component in a wide variety of industrial applications including pharmaceutical, food, leather and detergent formulation. Proteases are ubiquitous as they are found in plants, animals, fungi, bacteria and viruses. Microbial sources of proteases, particularly bacteria, are of particular interest in industry. The isolation and identification of protease producing bacteria, hence, is the most basic pre-requisite in this regard for the success of any industry based on protease utilization. After their isolation from different sources, bacteria are routinely identified by morphological and biochemical characterization assays. Newer molecular techniques now allow bacterial species to be identified with authenticity by their genetic sequences. Different sources of bacteria for protease production have been exploited but soil samples, tannery and food processing industries including kitchen waste forms the basis of this report, wherein the procedures and protocols for their isolation and identification are discussed. Key Words: Isolation, Protease, Identification, Biochemical characterization, Morphological assay
1.INTRODUCTION Proteases are a group of enzymes that catalyse the process of proteolysis in which proteins are broken down into single amino acids or smaller polypeptides. Proteases work to simplify proteins by disintegrating the peptide bonds by hydrolysis present within them. Protease enzymes also hold a pivotal role in biological functions, like the simplification of proteins into amino acids, their digestion followed by cell signaling (King et al., 2014; Shen et al., 2006). Proteases also cause a change by disrupting the function of a protein and breaking it down to its simpler components, which leads to the activation of a signal or a particular function in a biological pathway. Proteases occupy an important position with respect to applications in commercial as well as physiological industries. Proteases act as an integral component in a wide range of industrial applications including pharmaceutical, food, leather and detergent formulation (Mohen et al., 2005). Also, of all the total enzymes present globally, proteases account for two thirds and about quarter of the total global enzyme produced
© 2023, IRJET
|
Impact Factor value: 8.226
|
ISO 9001:2008 Certified Journal
|
Page 1131