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Indian Journal of Microbiology

, Volume 59, Issue 4, pp 436–444 | Cite as

Mutanase Enzyme from Paracoccus mutanolyticus RSP02: Characterization and Application as a Biocontrol Agent

  • Sudheer Kumar Buddana
  • Ravi Naga Amrutha
  • Uma Rajeswari Batchu
  • Suprasanna Penna
  • Reddy Shetty PrakashamEmail author
Original research article
  • 13 Downloads

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.

Keywords

Mutanase Enzyme kinetics Streptococcal biofilm Zymography Antifungal activity 

Notes

Acknowledgements

Authors are thankful to the Director, CSIR-IICT Hyderabad. Mr. Sudheer Kumar B gratefully acknowledges the CSIR, New Delhi, for providing Senior Research Fellowship. R. Naga Amrutha thanks the Department of Science and Technology (DST), New Delhi for financial support. Uma Rajeswari Batchu for BRNS, Mumbai for providing JRF. The manuscript communication number through CSIR- IICT is IICT/Pubs/2019/223.

Supplementary material

12088_2019_821_MOESM1_ESM.pptx (715 kb)
Supplementary material 1 (PPTX 714 kb)

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Copyright information

© Association of Microbiologists of India 2019

Authors and Affiliations

  1. 1.Medicinal Chemistry and BiotechnologyCSIR-Indian Institute of Chemical TechnologyHyderabadIndia
  2. 2.Academy of Scientific and Innovative Research (AcSIR)CSIR-Indian Institute of Chemical TechnologyHyderabadIndia
  3. 3.Nuclear Agriculture and Biotechnology DivisionBhabha Atomic Research Centre (BARC)MumbaiIndia

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