Acta Scientific Microbiology (ISSN: 2581-3226)

Research Article Volume 6 Issue 3

Impact of Substrate pH and Enzyme-Substrate Incubation Time on Protease from a New Halo-tolerant Bor S17B13 Found in the Mangroves of Western India

Priyanka Sawant and Jignasha Thumar*

Department of Microbiology and Biotechnology, Gujarat University, Ahmedabad, Gujarat, India

*Corresponding Author: Jignasha Thumar, Department of Microbiology and Biotechnology, Gujarat University, Ahmedabad, Gujarat, India.

Received: January 17, 2023; Published: February 02, 2023

Abstract

Since enzymes are bioactive chemicals, they are of interest to scientists all over the world. Demand for microbial enzymes is rising rapidly across a wide range of sectors as their importance in ensuring the security, sustainability and efficiency of biotech processes becomes more widely recognised. Media optimization is a powerful tool to enhance production capacity from biological sources (microorganisms). In this present study, samples were collected from mangrove-rich sources such as Borivali Monari Creek and Jhow Island, Maharashtra, western India. Samples collected were mangrove-associated soil, mangrove root, and sea water. The type of sampling was random. Samples were collected with prior permission from the mangrove cell maharashtra government. A total of 30 samples were processed, from which 62 isolates were obtained. These isolates were all screened for industrially important enzymes (protease, amylase, and cellulase), and the best protease producer was selected for further studies. The method of optimization was "ONE VARIABLE AT A TIME" (OVAT) ” and method is a well-known optimization technique. Optimization was carried out for parameters like inoculum size, pH of the growth medium, various carbon sources, varied nitrogen sources, and metal ions and cations. Casein was used as substrate, and casein was prepared in various pH buffers for the enzyme assay. At 5 minutes after incubation of enzyme substrate mixture, enzyme activity was at its peak, and substrate degradation in the medium was minimal as compare 65 minutes set. Enzyme activity decreased d ramatically as substrate pH increased (between 11 and 12 pH). This optimization supports maximizing the yield of protease production and is of great benefit to the biotech industry. The present study shows that with time stability of enzyme reduces and residues of degraded substrate increases within the reaction mixture. Here in our experiment variation in substrate pH were studied, which showed that protease from Bor S17B13 isolate has optimum enzyme activity at substrate pH 10 at highly alkaline pH then 10 pH, activity of enzyme was reduced. A phenomenon called dissociation of substrate (casein) was seen at casein solution 13 pH.

Keywords: Mangrove Ecosystem; Halotolerant; Protease; Bor S17B13; Substrate pH; Enzyme Substrate; ES

References

  1. Q K Beg and R Gupta. “Purification and characterization of an oxidation-stable, thiol-dependent serine alkaline protease from Bacillus mojavensis”. Enzyme and Microbial Technology2 (2003): 294-304.
  2. M K Panda., et al. “Isolation and characterization of a thermophilic Bacillus sp. with protease activity isolated from hot spring of Tarabalo, Odisha, India”. Iranian Journal of Microbiology2 (2013): 159-165.
  3. P Sawant and J Thumar. “Amalgamation of technologies and scientrepreneur prospective of electrochemically active halophilic microbugs - a review paper". Vidya-A journal of Gujarat Univeristy2 (2021): 125-156.
  4. B Johnvesly and G R Naik. “Studies on production of thermostable alkaline protease from thermophilic and alkaliphilic Bacillus JB-99 in a chemically defined medium”. Process Biochemistry 37.2 (2001): 139-144.
  5. U Jadhav and H Hocheng. “A review of recovery of metals from industrial waste”. Journal of Achievements in Materials and Manufacturing 54 (2012): 159-167.
  6. P Sawant and J Thumar. “Isolation of Marine Microbugs from Mangrove Ecosystem and Screening of Industrially Important Enzymes”. Acta Scientific Microbiology (2022): 35-39.
  7. R C Kasana., et al. “A rapid and easy method for the detection of microbial cellulases on agar plates using Gram’s iodine”. Current Microbiology5 (2008): 503-507.
  8. B O De Veras., et al. “Screening of protease, cellulase, amylase and xylanase from the salt-tolerant and thermostable marine Bacillus subtilis strain SR60”. F1000 Research 7 (2018): 1704.
  9. R Zalak., et al. “Isolation and Screening of Multivariant Enzyme Producing Endophytic Bacteria from Citrus limon”. Acta Scientific Microbiology 5 (2022): 2581-3226.
  10. Hagihara. “The Enzymes”. 2nd edn. vol. 4. Boyer “. P. Acad. Press New York, (1958).
  11. J T Thumar and S P Singh. “Secretion of an alkaline protease from a salt- tolerant and alkaliphilic, Streptomyces clavuligerus strain Mit-1”. Brazilian Journal of Microbiology4 (2007): 766-772.
  12. R K Patel., et al. “Production of Extracellular Halo-alkaline Protease from a Newly Isolated Haloalkaliphilic Bacillus sp. Isolated from Seawater in Western India”. World Journal of Microbiology and Biotechnology 4 (2006): 375-382.
  13. C Masi., et al. “Isolation, screening, characterization, and identification of alkaline protease-producing bacteria from leather industry effluent”. Annals of Microbiology 71.1 (2021): 1-11.
  14. L Meza-Espinoza., et al. “Enzyme activity and partial characterization of proteases obtained from Bromelia karatas fruit and compared with Bromelia pinguin proteases”. Food Science and Biotechnology 2 (2018): 509-517.
  15. M Amid., et al. “Purification and characterization of alkaline-thermostable protease enzyme from Pitaya (Hylocereus polyrhizus) waste: a potential low cost of the enzyme”. BioMed Research International (2014): 259238.
  16. N Abdullah Al-Dhabi., et al. “Characterization and fermentation optimization of novel thermo stable alkaline protease from Streptomyces Al-Dhabi-82 from the Saudi Arabian environment for eco-friendly and industrial applications”. Journal of King Saud University - Science 32.1 (2020): 1258-1264.
  17. Nadeem J., et al. “Purification and characterization of an alkaline protease from Bacillus licheniformis UV-9 for detergent formulations”. Songklanakarin Journal of Science and Technology (SJST) 35 (2013): 187-195.
  18. H Bian and J Plank. “Re-association Behavior of Casein Submicelles in Highly Alkaline Environments”. Zeitschrift für Naturforsch B 67 (2012).

Citation

Citation: Priyanka Sawant and Jignasha Thumar. “Impact of Substrate pH and Enzyme-Substrate Incubation Time on Protease from a New Halo-tolerant Bor S17B13 Found in the Mangroves of Western India". Acta Scientific Microbiology 6.2 (2023): 03-09.

Copyright

Copyright: © 2022 Priyanka Sawant and Jignasha Thumar. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.




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