Purification and Characterization of an Active Principle, Lawsone, Responsible for the Plasmid Curing Activity of Plumbago zeylanica Root Extracts
Plasmid curing is the process of obviating the plasmid encoded functions such as antibiotic resistance, virulence, degradation of aromatic compounds, etc. in bacteria. Several plasmid curing agents have been reported in literature, however, no plasmid curing agent can eliminate all plasmids from dif...
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doaj-cc0a61b0265849fb8b362d4e9492b3e62020-11-24T21:00:39ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2018-11-01910.3389/fmicb.2018.02618380787Purification and Characterization of an Active Principle, Lawsone, Responsible for the Plasmid Curing Activity of Plumbago zeylanica Root ExtractsRajashree Bhalchandra Patwardhan0Prashant Kamalakar Dhakephalkar1Balu Ananda Chopade2Dilip D. Dhavale3Ramesh R. Bhonde4Department of Microbiology, Haribhai V. Desai College of Arts, Science and Commerce, Pune University, Pune, IndiaBioenergy Group, Agharkar Research Institute, Pune, IndiaDepartment of Microbiology, Savitribai Phule Pune University, Aurangabad, IndiaDepartment of Chemistry, Savitribai Phule Pune University, Pune, IndiaSchool of Regenerative Medicine – Manipal Academy of Higher Education, Bengaluru, IndiaPlasmid curing is the process of obviating the plasmid encoded functions such as antibiotic resistance, virulence, degradation of aromatic compounds, etc. in bacteria. Several plasmid curing agents have been reported in literature, however, no plasmid curing agent can eliminate all plasmids from different hosts. Hence, there is always a need for novel plasmid curing agents that can be effectively used for reversal of plasmid encoded functions such as virulence, antibiotic resistance, etc. In the present study, an active principle responsible for the plasmid curing activity was purified from roots of Plumbago zeylanica by bioassay guided fractionation and identified as 2-hydroxy-1,4-naphthoquinone (lawsone), on the basis of spectral and analytical data such as NMR, GCMS, FTIR. Plasmid curing activity of lawsone was observed against reference as well as wild plasmids (pBR322, pRK2013, R136, pUPI281, and pUPI282) residing in a range of hosts. Curing of plasmid was confirmed by agarose gel electrophoresis. MICs of antibiotics against A. baumannii A24 (pUPI281) and E. coli (pRK2013) decreased significantly in presence of lawsone suggesting synergy between lawsone and antibiotics. Lawsone also inhibited transfer of plasmid pRK2013 to E. coli either by transformation or conjugation. Viability assays (MTT) revealed that lawsone was not toxic to mammalian cells. Thus, the present investigation has revealed lawsone as an effective plasmid curing agent capable of suppressing development and spread of antibiotic resistance. Further, lawsone has important application in basic research to identify phenotypes encoded by the plasmids in plasmid curing experiments. To the best of our knowledge this is the first report of plasmid curing activity of lawsone isolated from roots of P. zeylanica.https://www.frontiersin.org/article/10.3389/fmicb.2018.02618/fulllawsone(2-hydroxy-14 naphthoquinone)Plumbago zeylanicaextractionpurificationplasmid curing |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Rajashree Bhalchandra Patwardhan Prashant Kamalakar Dhakephalkar Balu Ananda Chopade Dilip D. Dhavale Ramesh R. Bhonde |
spellingShingle |
Rajashree Bhalchandra Patwardhan Prashant Kamalakar Dhakephalkar Balu Ananda Chopade Dilip D. Dhavale Ramesh R. Bhonde Purification and Characterization of an Active Principle, Lawsone, Responsible for the Plasmid Curing Activity of Plumbago zeylanica Root Extracts Frontiers in Microbiology lawsone(2-hydroxy-1 4 naphthoquinone) Plumbago zeylanica extraction purification plasmid curing |
author_facet |
Rajashree Bhalchandra Patwardhan Prashant Kamalakar Dhakephalkar Balu Ananda Chopade Dilip D. Dhavale Ramesh R. Bhonde |
author_sort |
Rajashree Bhalchandra Patwardhan |
title |
Purification and Characterization of an Active Principle, Lawsone, Responsible for the Plasmid Curing Activity of Plumbago zeylanica Root Extracts |
title_short |
Purification and Characterization of an Active Principle, Lawsone, Responsible for the Plasmid Curing Activity of Plumbago zeylanica Root Extracts |
title_full |
Purification and Characterization of an Active Principle, Lawsone, Responsible for the Plasmid Curing Activity of Plumbago zeylanica Root Extracts |
title_fullStr |
Purification and Characterization of an Active Principle, Lawsone, Responsible for the Plasmid Curing Activity of Plumbago zeylanica Root Extracts |
title_full_unstemmed |
Purification and Characterization of an Active Principle, Lawsone, Responsible for the Plasmid Curing Activity of Plumbago zeylanica Root Extracts |
title_sort |
purification and characterization of an active principle, lawsone, responsible for the plasmid curing activity of plumbago zeylanica root extracts |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Microbiology |
issn |
1664-302X |
publishDate |
2018-11-01 |
description |
Plasmid curing is the process of obviating the plasmid encoded functions such as antibiotic resistance, virulence, degradation of aromatic compounds, etc. in bacteria. Several plasmid curing agents have been reported in literature, however, no plasmid curing agent can eliminate all plasmids from different hosts. Hence, there is always a need for novel plasmid curing agents that can be effectively used for reversal of plasmid encoded functions such as virulence, antibiotic resistance, etc. In the present study, an active principle responsible for the plasmid curing activity was purified from roots of Plumbago zeylanica by bioassay guided fractionation and identified as 2-hydroxy-1,4-naphthoquinone (lawsone), on the basis of spectral and analytical data such as NMR, GCMS, FTIR. Plasmid curing activity of lawsone was observed against reference as well as wild plasmids (pBR322, pRK2013, R136, pUPI281, and pUPI282) residing in a range of hosts. Curing of plasmid was confirmed by agarose gel electrophoresis. MICs of antibiotics against A. baumannii A24 (pUPI281) and E. coli (pRK2013) decreased significantly in presence of lawsone suggesting synergy between lawsone and antibiotics. Lawsone also inhibited transfer of plasmid pRK2013 to E. coli either by transformation or conjugation. Viability assays (MTT) revealed that lawsone was not toxic to mammalian cells. Thus, the present investigation has revealed lawsone as an effective plasmid curing agent capable of suppressing development and spread of antibiotic resistance. Further, lawsone has important application in basic research to identify phenotypes encoded by the plasmids in plasmid curing experiments. To the best of our knowledge this is the first report of plasmid curing activity of lawsone isolated from roots of P. zeylanica. |
topic |
lawsone(2-hydroxy-1 4 naphthoquinone) Plumbago zeylanica extraction purification plasmid curing |
url |
https://www.frontiersin.org/article/10.3389/fmicb.2018.02618/full |
work_keys_str_mv |
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