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Modified two-phase system for partition of Bacillus macerans spores
Abstract:
An aqueous two-phase system made with polyethylene glycol (PEG) 1000 and potassium phosphate gives much higher recovery of Bacillus macerans spores in the upper phase (PEG rich) than does a similar system utilizing PEG 4000. The upper phase completely rejects vegetative cells, which collect at the interface. The system may be useful in purifying spores of other species. Scanning electron micrographs of B. macerans spores cleaned in this system show no obvious attached sporangial fragments. The micrographs show that the ridged coat may form polygonal structures at the poles, as previously observed in Bacillus polymyxa spores.
Insights
Polyethylene glycol (PEG) 1000 and potassium phosphate systems efficiently recover Bacillus macerans spores. This aqueous two-phase system also purifies spores by separating them from vegetative cells.
Area of Science:
- Microbiology
- Biochemistry
- Separation Science
Background:
- Bacillus macerans spores are of interest for various applications.
- Efficient separation and purification of bacterial spores are crucial for research and industrial processes.
- Aqueous two-phase systems (ATPS) offer a promising method for bioparticle separation.
Purpose of the Study:
- To evaluate the effectiveness of an ATPS using polyethylene glycol (PEG) 1000 and potassium phosphate for Bacillus macerans spore recovery.
- To compare the recovery efficiency of PEG 1000-based ATPS with PEG 4000-based ATPS.
- To assess the purity of recovered spores and investigate spore coat morphology.
Main Methods:
- Preparation of an aqueous two-phase system using polyethylene glycol (PEG) 1000 and potassium phosphate.
- Separation of Bacillus macerans spores and vegetative cells within the ATPS.
- Analysis of spore recovery in the upper PEG-rich phase.
- Scanning electron microscopy (SEM) of purified spores to examine surface morphology.
Main Results:
- The PEG 1000/potassium phosphate system demonstrated significantly higher recovery of Bacillus macerans spores compared to a PEG 4000 system.
- Vegetative cells were effectively excluded from the upper phase, accumulating at the interface.
- Scanning electron micrographs revealed purified spores with no significant sporangial fragments and showed polygonal structures on the ridged coat.
Conclusions:
- An aqueous two-phase system with PEG 1000 and potassium phosphate is highly effective for recovering Bacillus macerans spores.
- This ATPS provides a method for purifying spores by separating them from vegetative cells.
- The system shows potential for purifying spores of other bacterial species and aids in studying spore coat structures.