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Updated: Jun 30, 2026
Purification of a High Molecular Mass Protein in Streptococcus mutans
Published on: September 14, 2019
Methods for obtaining active mutanase preparations from Paenibacillus curdlanolyticus
Małgorzata Pleszczyńska1, Adrian Wiater, Janusz Szczodrak
1Department of Industrial Microbiology, Maria Curie-Skłodowska University, Lublin, Poland. mplesz@poczta.onet.pl
Abstract:
The strain Paenibacillus curdlanolyticus MP-1 was used to obtain mutan-hydrolyzing enzymes. Different methods of precipitation and concentration of the post culture liquid were tested. All these methods produced satisfactory results in regard to the overall activity of mutanase and yielded active preparations of the enzyme. The best precipitation was obtained with propanol -98% of the initial enzyme activity was preserved with a purification of 2-fold. Salting out with ammonium sulfate at 50% saturation gave mutanase recovery of 77% and a purification of around 2-fold. Ultrafiltration yielded an about 10-fold concentrated preparation of the enzyme with a yield of 98%. Lyophilization and concentration of the culture broth (in the range from 5 to 20 times) in a vacuum evaporator yielded active crude preparations with mutanase recovery of 97%.
Insights
Researchers optimized methods for isolating mutanase enzymes from Paenibacillus curdlanolyticus MP-1. Propanol precipitation and ultrafiltration proved effective, preserving high enzyme activity and achieving significant purification for biotechnological applications.
Area of Science:
- Enzymology
- Biotechnology
- Microbiology
Background:
- Mutanase enzymes are crucial for hydrolyzing mutan, a component of dental plaque.
- Efficient isolation and purification of active mutanase are essential for its industrial and therapeutic applications.
Purpose of the Study:
- To evaluate different precipitation and concentration techniques for isolating mutan-hydrolyzing enzymes.
- To identify the most effective method for obtaining active and purified mutanase preparations from Paenibacillus curdlanolyticus MP-1.
Main Methods:
- Tested precipitation methods including propanol and ammonium sulfate salting out.
- Evaluated concentration techniques such as ultrafiltration and vacuum evaporation (lyophilization).
- Assessed enzyme activity, recovery, and purification fold for each method.
Main Results:
- Propanol precipitation preserved 98% of initial enzyme activity with 2-fold purification.
- Ultrafiltration achieved a 10-fold concentration with 98% yield.
- Vacuum evaporation yielded active crude preparations with 97% mutanase recovery.
Conclusions:
- Multiple methods effectively yielded active mutanase preparations.
- Propanol precipitation and ultrafiltration are highly efficient for mutanase isolation and concentration.
- Optimized enzyme recovery and purification facilitate potential applications in dental health and biotechnology.
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