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Switchable deep eutectic solvent driven micellar extractive fermentation of ultrapure fibrin digesting enzyme from
Ramya Muniasamy1, Bhavani Sowndharya Balamurugan1, Devi Rajamahendran1
1Green Separation Engineering Laboratory, School of Chemical and Biotechnology, SASTRA Deemed To Be University, Thanjavur, Tamil Nadu, 613401, India.
Scientific Reports
|January 19, 2022
Summary
This study introduces pH-driven micellar two-phase extraction for producing and purifying fibrinolytic protease (FLP) from Bacillus subtilis using sustainable shrimp waste. The novel method achieved high enzyme purity and activity, demonstrating effective fibrinolytic potential.
Area of Science:
- Biotechnology
- Enzyme Engineering
- Biochemistry
Background:
- Fibrinolytic protease (FLP) is crucial for treating thrombolytic diseases.
- Current production and purification methods can be inefficient.
- Sustainable and cost-effective enzyme production is highly desirable.
Purpose of the Study:
- To develop a pH-driven swappable micellar two-phase extraction for concurrent production and purification of FLP.
- To utilize sustainable shrimp waste as a low-cost substrate for FLP production.
- To optimize and validate the extraction and purification process for high-purity FLP.
Main Methods:
- Extractive fermentation using a pH swap mechanism with surfactant deep eutectic solvents (SDES).
- Optimization of extractive fermentation using Response Surface Methodology (RSM).
- Purification of FLP using gel filtration chromatography and anion exchange chromatography.
Main Results:
- Maximum protease yield of 185 U/mg from shrimp waste.
- Optimized enzyme activity of 248 IU/mg using RSM.
- Achieved maximum purity fold of 22.32 with enzyme activity of 1172 IU/ml.
- Confirmed in-vitro fibrinolytic activity via fibrin plate assay.
Conclusions:
- pH-driven micellar two-phase extraction is an effective strategy for concurrent FLP production and purification.
- Surfactant deep eutectic solvents facilitate efficient extraction of FLP.
- The developed method offers a sustainable and scalable approach for therapeutic enzyme production.
