ORIGINAL RESEARCH
Whey Fermentation for Protease Production Using
Bacillus thuringiensis Isolated from Mangrove
Rhizosphere Soil in Jazan, Saudi Arabia
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1
Biology Department, Faculty of Science, Jazan University, Jazan, Kingdom of Saudi Arabia
2
The Holding Company for Biological Products and Vaccines, Vacsera, Cairo, Egypt
3
Botany Department, Faculty of Science, Fayoum University, Fayoum, Egypt
4
Botany and Microbiology Department, Faculty of Science, Helwan University, Cairo, Egypt
Submission date: 2019-05-12
Final revision date: 2019-07-06
Acceptance date: 2019-07-09
Online publication date: 2020-02-05
Publication date: 2020-03-31
Corresponding author
Khaled E. El-Gayar
Biology Department- Faculty of Science- Jazan University-KSA, Biology Department- Faculty of Science- Jazan Univ, 45142, Jazan, Saudi Arabia
Pol. J. Environ. Stud. 2020;29(3):2167-2176
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ABSTRACT
Utilization of whey as a fermentation substrate offers an opportunity to produce valuable products.
The current work was carried out in order to evaluate the capability of bacteria isolated from mangrove
rhizosphere soil in Jazan (Kingdom of Saudi Arabia) for the production of extracellular protease using
fresh whey as a substrate. The bacterial isolate with the maximum proteolytic activity was identified
by biochemical and 16S rDNA techniques as Bacillus thuringiensis. The factors affecting bacterial
growth and protease production were investigated. The maximum protease production was achieved
with minimal medium supplemented with 50% whey, 1% skim milk, 10% NaCl, and 1% CaCl2 at pH
7.0 and temperature 37ºC. In a fermentation experiment, B. thuringiensis utilized whey as a substrate
for the production of extracellular protease with a high specific activity. The protease was precipitated
using ammonium sulfate, followed by dialysis to yield 9.7-fold purification with 70.8% recovery. After
fermentation, there was a remarkable reduction of BOD, COD, and COD/BOD ratio. A high yield of
biomass with a promising protein level was obtained. In conclusion, B. thuringiensis demonstrated
a high capability for extracellular protease and single cell protein production using whey as a costeffective
medium that could minimize environmental pollution.