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Research Article Open access CC BY 4.0

Protease Production by Submerged Fermentation in Shake Flasks Using Bacillus sp. Isolated from the Soil

J. Okpalla, D. A. Onyekuru, I. E. Duru, T. O. Mba

Microbiology Research Journal International · pp. 42–51 · Published 26 Dec 2022

10.9734/mrji/2022/v32i101350

Abstract

Proteases are one of the most industrially important enzymes, which account for about 60% of total enzyme market. Protease production by submerged fermentation in shake flasks using Bacillus sp. isolated from the soil was studied. Soil samples were collected from different locations within Chukwuemeka Odumegwu Ojukwu University, Uli, Anambra state. The soil samples were serially diluted and inoculated on sterilized skim milk agar plates. The plates were incubated at 30oC for 72 h. A clear zone around the colonies gave an indication of protease-producing bacteria isolates.   The selected protease producers were subsequently used for shake flask fermentation in 50 ml sterile medium. Optimization study was conducted to determine the effect of carbon sources, nitrogen sources, trace elements, agitation rates and pH.  Twenty one bacteria isolates were found to be active protease producers and isolates RS-5 and OS-9 had the highest zone of clearance of 13.5 and 12.1 mm respectively. The result of submerged production of protease by the  bacteria isolates  revealed that the isolates RS-5 and OS-9 accumulated maximum protease yield of 3.23 and 2.71 U/ml respectively. The isolates were Gram positive and endospore formers, and were identified as Bacillus sp. RS-5 and OS-9.The addition of Starch and maltose stimulated optimum protease production of 3.47 and 2.77 U/ml by Bacillus sp. RS-5 and OS-9 respectively. Beef extract enhanced maximum enzyme yield of 3.35 and 2.90 U/ml for Bacillus sp. RS-5 and OS-9 respectively. Maximum protease yield of 3.28 U/ml for Bacillus sp. RS-5 and 2.85 U/ml for Bacillus sp. OS-9 was obtained by the supplementation of 0.4 g/l of FeS04 respectively. The maximum protease yield was observed at agitation rate of 200 rpm for Bacillus sp. RS-5 and 170 rpm for Bacillus sp. OS-9. At pH8, protease accumulation was highest for Bacillus sp. RS-5 and OS-9. The study revealed that the soil harbours some protease-producing bacteria strains and protease production can be greatly enhanced through optimization of process parameters.

Soil Bacillus species protease submerged fermentation trace elements

References (19)

  1. 1 Purification and characterization of an oxidation-stable, thiol-dependent serine alkaline protease from Bacillus mojavensis [DOI]
  2. 2 Production of protease from a new alkalophilic Bacillus sp. I-312 grown on soybean meal: optimization and some properties [DOI]
  3. 3 Optimization of physical factors affecting the production of thermo-stable organic solvent-tolerant protease from a newly isolated halo tolerant Bacillus subtilis strain Rand [DOI]
  4. 4 Production of Protease by Bacillus sp. N-40 Isolated from Soil and Its Enzymatic Properties
  5. 5 Alkaline protease production by submerged fermentation in stirred tank reactor using Bacillus licheniformis NCIM-2042: Effect of aeration and agitation regimes [DOI]
  6. 6 Production dynamics of extracellular protease from Bacillus species [DOI]
  7. 7 The production of alkaline protease by a Bacillus species isolate [DOI]
  8. 8 Optimization of thermostable alkaline protease production from species of Bacillus using rice bran [DOI]
  9. 9 Optimal conditions for production of extracellular protease from newly isolated Bacillus cereus strain CA15 [DOI]
  10. 10 Optimization of environmental and nutritional conditions for the production of alkaline protease by a newly isolated bacterium Bacillus cereus strain 146
  11. 11 Optimization of alkaline protease production from Pseudomonas fluorescens isolated from meat waste contaminated soil [DOI]
  12. 12 Isolation, purification and mass production of protease enzyme from bacillus subtilis
  13. 13 Protease obtention using Bacillus subtilis 3411 and amaranth seed meal medium at different aeration rates [DOI]
  14. 14 Production of alkaline protease in a low-cost medium by alkalophilic Bacillus sp. and properties of the enzyme [DOI]
  15. 15 Laundry Performance Of Subtilisin Proteases [DOI]
  16. 16 The benefit of detergent enzymes under changing washing conditions [DOI]
  17. 17 Screening and Optimal Protease Production by Bacillus sp. Sw-2 Using Low Cost Substrate Medium [DOI]
  18. 18 Purification and Characterization of Caseinolytic Extracellular protease from Bacillus amyloliquefaciens S94
  19. 19 Studies on the production of lipase from recombinant Staphylococcus carnosus [DOI]

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