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Biodegradation of chlorpropham and its major products by Bacillus licheniformis NKC-1

  • Namadev K. Pujar
  • H. G. Premakshi
  • Shruti Laad
  • Shridhar V. Pattar
  • Manisha Mirjankar
  • Chandrappa M. Kamanavalli
Original Paper

Abstract

Chlorpropham [isopropyl N-(3-chlorophenyl) carbamate] (CIPC), an important phenyl carbamate herbicide, has been used as a plant growth regulator and potato sprout suppressant (Solanum tuberosum L) during long-term storage. A bacterium capable of utilizing the residual herbicide CIPC as a sole source of carbon and energy was isolated from herbicide-contaminated soil samples employing selective enrichment method. The isolated bacterial strain was identified as Bacillus licheniformis NKC-1 on the basis of its morphological, cultural, biochemical characteristics and also by phylogenetic analysis based on 16S rRNA gene sequences. The organism degraded CIPC through its initial hydrolysis by CIPC hydrolase enzyme to yield 3-chloroaniline (3-CA) as a major metabolic product. An inducible 3-CA dioxygenase not only catalyzes the incorporation of molecular oxygen but also removes the amino group by the deamination yielding a monochlorinated catechol. Further, degradation of 4-chlorocatechol proceeded via ortho- ring cleavage through the maleylacetate process. 3-Chloroaniline and 4-chlorocatechol are the intermediates in the CIPC degradation which suggested that dechlorination had occurred after the aromatic ring cleavage. The presence of these metabolites has been confirmed by using ultra-violet (UV), high-performance liquid chromatography (HPLC), thin layer chromatography (TLC), Fourier transmission-infrared (FT-IR), proton nuclear magnetic resonance (1H NMR) and gas chromatography-mass (GC-MS) spectral analysis. Enzyme activities of CIPC hydrolase, 3-CA dioxygenase and chlorocatechol 1, 2-dioxygenase were detected in the cell-free-extract of the CIPC culture and are induced by cells of NKC-1 strain. These results demonstrate the biodegradation pathways of herbicide CIPC and promote the potential use of NKC-1 strain to bioremediate CIPC-contaminated environment with subsequent release of ammonia, chloride ions and carbon dioxide.

Keywords

Biodegradation Chlorpropham B. licheniformis NKC-1 Sprout suppressant GC-MS spectral analysis 

Notes

Acknowledgements

Authors are thankful to the DST, New Delhi, for providing financial assistance (Grant No. PURSE-Phase-2/3 (G), SR).

Supplementary material

11274_2018_2494_MOESM1_ESM.docx (512 kb)
Supplementary material 1 (DOCX 512 KB)

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Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  • Namadev K. Pujar
    • 1
  • H. G. Premakshi
    • 2
  • Shruti Laad
    • 1
  • Shridhar V. Pattar
    • 1
  • Manisha Mirjankar
    • 1
  • Chandrappa M. Kamanavalli
    • 1
  1. 1.P. G. Department of Studies in BiochemistryKarnatak UniversityDharwadIndia
  2. 2.P. G. Department of Studies in ChemistryKarnatak UniversityDharwadIndia

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