Abstract
Green chemistry technologies are the powerful tool for the management of environmental wastes challenges. Agro industrial residues are composed of complex polysaccharides that support the microbial growth for the production of useful products (enzymes, organic acids, drugs, etc.). Disposal and environment friendly management of these wastes has become a global priority. The aim of present investigation was to improve the alkaline protease yield by treating the parent Bacillus subtilis M-9 strain with different mutagens UV-irradiations, N-methyl-N-nitro- N-nitrosoguinidine (NTG), Ethidium bromide (EB), using agroindustrial wastes (banana stalk and corn stover) in submerged fermentation. Fifteen positive mutants were selected on skim milk agar plates for shake flask experiments. BSU-5 mutant strain showed 81.21± 3.24 PU/mL alkaline protease activity higher than parent strain (23.57 ± 1.19 PU/mL) in optimized fermentation medium. The fermentation profile like pH (9), temperature (45°C), inoculum size (2 mL), incubation time (24 hrs, and kinetic parameters such as u (h-1), Yp/s, Yp/x, Yx/s, qs, Qs, qp also confirmed the hyper proteolytic activity of alkaline protease produced from BSU-5 mutant strain over parent strain and other mutants. Finally, the BSU-5 mutant strain was immobilized by entrapping it in calcium alginate beads and agar. Alkaline protease production and stability of biocatalyst were investigated in both free and immobilized cells. It was concluded from the study, immobilized cells were more efficient for enzyme production then free cells when used repeatedly.
Highlights
Synthetic and expensive substrates are being replaced by agroindustrial byproducts for the production of a wide range of value added biotechnological products [1,2]
The parent strain B. subtilis M-9 was treated with different mutagens like UV irradiation, NTG (N-methyl-N-nitro-N-nitrosoguinidine) and EB, to improve its productivity
The culture of Bacillus subtilis M-9 was treated with NTG and EB for different time intervals. 22.3% survival rate was observed with NTG after an exposure of 140 minutes (Table 1)
Summary
Synthetic and expensive substrates are being replaced by agroindustrial byproducts for the production of a wide range of value added biotechnological products [1,2]. Lignocellulosic biomass comprising forestry, agricultural and agro industrial wastes are abundant, renewable and inexpensive energy sources. Such wastes include a variety of materials such as sawdust, poplar trees, sugarcane bagasse and straws, stems, stalks, leaves, husks, shells and peels from cereals like rice, wheat, corn. Lignocelluloses wastes are accumulated every year in large quantities, causing environmental problems. Due to their chemical composition based on sugars and other compounds of interest, they could be utilized for the production of a number of value added products, such as ethanol, food additives, organic acids, enzymes and others. Besides the environmental problems caused by their accumulation in the nature, the non use of these materials constitutes a loss of potentially valuable sources [3]
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