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Updated: Jan 3, 2026
Purification of a High Molecular Mass Protein in Streptococcus mutans
Published on: September 14, 2019
Mutanase Enzyme from Paracoccus mutanolyticus RSP02: Characterization and Application as a Biocontrol Agent
Sudheer Kumar Buddana1,2, Ravi Naga Amrutha1, Uma Rajeswari Batchu1
11Medicinal Chemistry and Biotechnology, CSIR-Indian Institute of Chemical Technology, Hyderabad, 500 007 India.
Abstract:
Mutanases are enzymes that have the ability to cleave α-1,3 linkages in glucan polymer. In the present investigation, mutanase enzyme purified from the culture filtrate of Paracoccus mutanolyticus was evaluated for Streptococcal biofilm degradation and antimicrobial activity against pathogenic fungi along with enzyme kinetics, activation energies, pH and thermal stability. Biochemical and molecular characterization depicted that the enzyme showed optimum activity at pH 5.5 and at 50 °C. It displayed Michaelis-Menten behaviour with a Km of 1.263 ± 0.03 (mg/ml), Vmax of 2.712 ± 0.15 U/mg protein. Thermal stability studies denoted that it required 55.46 and 135.43 kJ mol-1 of energy for activation and deactivation in the temperature range of 30-50 °C and 50-70 °C respectively. Mutanase activity was enhanced ~ 50 and 75% by Fe2+ and EDTA, respectively, while presence of Hg2+ and Mn2+ inhibit > 90% of its activity. This enzyme has a molecular mass of 138 kDa and showed monomeric nature by Zymography. Scanning electron microscopy analysis of mutanase treated Streptococcal cells revealed cleavage of linkages among the cells and complete separation of cells, indicating its potential in dentistry as an anticaries agent in the prophylaxis and therapy of dental caries. In addition, antifungal activity of mutanase against Colletotrichum capsici MTCC 10147 and Cladosporium cladosporioide MTCC 7371 revealed that the enzyme has potential towards biological control of phytopathogens which could be used as an alternative bio-control agent against chemical pesticides in the future.
Insights
This study purified mutanase from Paracoccus mutanolyticus, demonstrating its effectiveness in degrading Streptococcus biofilms and inhibiting fungal growth. The enzyme shows potential as an anticaries agent and a bio-control agent for phytopathogens.
Area of Science:
- Enzymology
- Microbiology
- Biotechnology
Background:
- Mutanases are enzymes capable of cleaving α-1,3 linkages in glucan polymers.
- Dental caries are primarily caused by Streptococcus species, which produce biofilms.
- Phytopathogenic fungi pose significant threats to agriculture, necessitating novel biocontrol agents.
Purpose of the Study:
- To purify and characterize mutanase from Paracoccus mutanolyticus.
- To evaluate the enzyme's efficacy against Streptococcus biofilms and pathogenic fungi.
- To determine the enzyme's kinetic properties, stability, and potential applications.
Main Methods:
- Enzyme purification from Paracoccus mutanolyticus culture filtrate.
- Biochemical and molecular characterization including pH, temperature, and kinetic studies.
- Scanning electron microscopy for biofilm degradation analysis.
- Antifungal activity assays against Colletotrichum capsici and Cladosporium cladosporioide.
Main Results:
- Optimized enzyme activity at pH 5.5 and 50°C, with Michaelis-Menten kinetics (Km = 1.263 mg/ml, Vmax = 2.712 U/mg).
- Demonstrated significant degradation of Streptococcus biofilms and potent antifungal activity.
- Identified Fe2+ and EDTA as activators, while Hg2+ and Mn2+ inhibited activity.
Conclusions:
- Mutanase from P. mutanolyticus effectively degrades Streptococcus biofilms, indicating potential as an anticaries agent.
- The enzyme exhibits antifungal properties against key phytopathogens, suggesting its use as a bio-control agent.
- This research highlights mutanase as a promising biocatalyst for applications in dentistry and agriculture.
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