Proposal

PROTEIN PROFILING AND ENZYMATIC STUDY OF PESTICIDE
DEGRADING BACTERIA
Research proposal
By
Chinmay Rout
M.Sc. Microbiology
Research advisor : Dr Jyoti R Rout
SCHOOL OF BIOLOGICAL SCIENCES ,AIPH UNIVERSITY, BBSR

 INTRODUCTION:
 Pesticides are chemical substances having biocidal properties .
 They are plant protection agent as they are used to prevent plants from adverse and
damaging effect of weeds, insects ,pests etc.
 Organophosphorus compounds, namely, glyphosate, chlorpyrifos, parathion, methyl

parathion, diazinon and phorate have been extensively used as pesticides .
 Organophosphate pesticides include heterogenous compound, such as

monocrotophos, phosphoric acid, and monocron, that have a phosphoric acid
derivatives, chemical structure, exposure to which contaminates a wide range of
water and terrestrial ecosystem .
 Several physical and chemical methods are there for the degradation of the
xenobiotic compound but the biological methods for degradation of xenobiotic
compound by exploiting the microbial metabolism would be the best approach .
 In the process of biodegradation microbial species also show other symptomatic
response to different environmental stresses .
 The proteomic analysis offers the crosslink between presence of actual stress
responsive genes, its product and reflects the protein profile of a cell under the
stress conditions .
 Overall protein content of a cell may get increased indicating the stimulation in the
expression pattern of stress responsive gene thereby fostering bacteria to tolerate
pesticide toxicity .
 REVIEW OF LITERATURE
AUTHOR/YEAR TITLE SOURCE FINDINGS
(Pankaj et al., Differential expression and Cross Mark The protein sample of the identified bacteria,
2016) characterization of cypermethrin- Bacillus thuringiensis, exposed to
degrading potential proteins in cypermethrin were subjected to two-
Bacillus thuringiensis. dimensional gel electrophoresis and 450
protein spots were reproducibly detected ,
quantitative analysis revealed the presence of
51 protein spots showing significant and
reproducible changes in protein expression
under pesticide stress.
(Aswathi et al., 2021) Chlorpyrifos induced ELSVIER Sixty-four out of 794 significantly up-regulated proteins
proteome remodelling of were specifically involved in carbohydrate metabolism,
Pseudomonas nitroreducens environmental and genetic information processing and
AR-3 potentially aid efficient biosynthesis of secondary metabolites. A presumptive
degradation of the pesticide . MBL-fold metallo-hydrolase protein showing 44%
sequence similarity to an organophosphate hydrolase
was identified from the genome
(Shweta et al., 2023) Chlorpyrifos mediated Research Here the overall findings revealed that Bacillus
amendment in protein Gate megaterium expressed a few enzymes/or protein in
profiling of Bacillus spp . order to show adaptation towards surrounding
environment . It can be an efficient degrader of
chlorpyrifos, hence, could be exploited for remediation
of chlorpyrifos contaminated sites .
 RATIONALE :
 To understand the molecular mechanisms and pathways involved in the

biodegradation of pesticides and to identify the key enzymes and proteins that are
responsible for the process .
 Protein profiling and enzymatic study can also help to discover novel genes and
enzymes that can be used for bioremediation of pesticide-contaminated soil and
water.
 METHODOLOGY :
 Isolation and identification of pesticide degrading bacteria .
 Collecting soil sample from pesticide contaminated site .
 Enriching them in a suitable medium containing the pesticide .
 Isolating the pure bacterial cultures on agar plate .
 Identifying them by morphological and molecular methods ,such as Gram
staining ,PCR amplification
 Protein extraction and quantification .
 Lysing the bacterial cells and extracting the total protein content using various
physical and chemical method .
 Protein profiling by SDS-PAGE
 Solubilized proteins will be subjected to SDS-PAGE in gel slab .
 Electrophoresis will be performed in a discontinuous buffer system ,then the gel will
be stained by Coomassie brilliant blue , then the profile of the protein will be
analysed .
 Screening of industrially important enzymes .
 Characterization of enzymes .
 SIGNIFICANCE IN PUBLIC HEALTH .
 Understand the molecular mechanisms and genetic diversity of these bacteria ,

which can enhance their bioremediation potential and efficiency .
 Identify and characterize the enzymes which are involved in pesticide degradation,
which can be used as biocatalysts for detoxifying pesticide residues in the
environment .
 Develop bioindicators for monitoring pesticide contamination and biodegradation in

soil and water .
REFERENCES
• Pankaj, Negi G, Gangola S, Khati P, Kumar G, Srivastava A, et al. Differential

expression and characterization of cypermethrin-degrading potential proteins in
Bacillus thuringiensis strain, SG4. 3 Biotech 2016;6:225.
https://doi.org/10.1007/s13205-016-0541-4.
• Aswathi A, Pandey A, Madhavan A, Sukumaran RK. Chlorpyrifos induced
proteome remodelling of Pseudomonas nitroreducens AR-3 potentially aid
efficient degradation of the pesticide. Environmental Technology & Innovation
2021;21:101307. https://doi.org/10.1016/j.eti.2020.101307.
• Shweta N, Kamal J, Sahu K. Chlorpyrifos Mediated Amendment in Protein
Profiling of Bacillus spp 2023;36:1–11.
https://doi.org/10.52228/JRUB.2023-36-1-1.
THANK YOU

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PROTEIN PROFILING AND ENZYMATIC STUDY OF PESTICIDE

SCHOOL OF BIOLOGICAL SCIENCES ,AIPH UNIVERSITY, BBSR

 Organophosphorus compounds, namely, glyphosate, chlorpyrifos, parathion, methyl

 Organophosphate pesticides include heterogenous compound, such as

 To understand the molecular mechanisms and pathways involved in the

 SIGNIFICANCE IN PUBLIC HEALTH .

 Understand the molecular mechanisms and genetic diversity of these bacteria ,

 Develop bioindicators for monitoring pesticide contamination and biodegradation in

• Pankaj, Negi G, Gangola S, Khati P, Kumar G, Srivastava A, et al. Differential

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