Isolation and Protein Characterization of Lindane Degrading Root Epiphytic Bacterium Arthrobacter sp. T16 from Typha latifolia

Tanvi SINGH, Dileep K. SINGH


Lindane, extensively used as pesticide, causes severe environmental hazard and is a threat to the humanity. The present study aims to assess the capability and mechanism of root epiphytic bacteria of wetland plant Typha latifolia to degrade lindane. Isolation of lindane degrading root epiphytic bacteria was done by standard enrichment technique and lindane degradation analysis was done using Gas Liquid Chromatography. Bacterial strain Arthrobacter sp. T16 was isolated and identified, which showed maximum degradation of 71.2 ± 1.3% of 50 mg l-1 lindane. Lindane biodegradation was accompanied with decrease in pH, increase in chloride ions concentration of culture medium and a positive dechlorination assay. Biodegradation potential of Arthrobacter sp. T16 was also studied at different lindane concentrations. Maximum degradation was observed at 10 mg l-1 lindane followed by 50 mg l-1 and 100 mg l-1 lindane. Lindane biodegradation kinetics study inferred that the average rate of lindane degradation increased with increase in lindane concentration. Lindane induced proteins in Arthrobacter sp. T16 were studied by SDS-PAGE. Distinctive polypeptides came into view in the presence of lindane and were identified as putative ABC transporter periplasmic amino acid-binding protein, elongation factor Tu and trifunctional transcriptional regulator/proline dehydrogenase/pyrroline-5-carboxylate dehydrogenase, each expressed due to lindane stress. This study specifies the potential of phytoremediation in controlling the environmental contamination problem with the help of indigenous organisms present in roots of plants.



accession number; bacterial suspension; half-life; mesocosm; pellet

Full Text:



Abhilash PC, Srivastava S, Srivastava P, Singh B, Jafri A, Singh N (2011). Influence of rhizospheric microbial inoculation and tolerant plant species on the rhizoremediation of lindane. Environmental and Experimental Botany 74:127-130.

Alvarez A, Benimeli CS, Saez JM, Fuentes MS, Cuozzo SA, Polti MA, Amoroso MJ (2012a). Bacterial bio-resources for remediation of hexachlorocyclohexane. International Journal of Molecular Sciences 13(11):15086-15106.

Alvarez A, Yañez ML, Benimeli CS, Amoroso MJ (2012b). Maize plants (Zea mays) root exudates enhance lindane removal by native Streptomyces strains. International Biodeterioration and Biodegradation 66(1):14-18.

Benimeli CS, Castro G, Chaile A, Amoroso MJ (2007). Lindane uptake and degradation by aquatic Streptomyces sp. strain M7. International Biodeterioration and Biodegradation 59(2):148-155.

Benimeli CS, Fuentes MS, Abate CM, Amoroso MJ (2008). Bioremediation of lindane-contaminated soil by Streptomyces sp. M7 and its effects on Zea mays growth. International Biodeterioration and Biodegradation 61(3):233-239.

Benndorf D, Davidson I, Babel W (2004). Regulation of catabolic enzymes during long-term exposure of Delftia acidovorans MC1 to chlorophenoxy herbicides. Microbiology 150(4):1005-1014.

Böltner D, Godoy P, Muñoz‐Rojas J, Duque E, Moreno‐Morillas S, Sánchez L, Ramos JL (2008). Rhizoremediation of lindane by root‐colonizing Sphingomonas. Microbial Biotechnology 1(1):87-93.

Caldas T, El Yaagoubi A, Richarme G (1998). Chaperone properties of bacterial elongation factor EF-Tu. Journal of Biological Chemistry 273(19):11478-11482.

Camacho-Pérez B, Ríos-Leal E, Rinderknecht-Seijas N, Poggi-Varaldo HM (2012). Enzymes involved in the biodegradation of hexachlorocyclohexane: a mini review. Journal of Environmental Management 95:S306-S318.

Cuozzo SA, Rollán GG, Abate CM, Amoroso MJ (2009). Specific dechlorinase activity in lindane degradation by Streptomyces sp. M7. World Journal of Microbiology & Biotechnology 25(9):1539-1546.

Datta J, Maiti AK, Modak DP, Chakrabartty PK, Bhattacharyya P, Ray PK (2000). Metabolism of γ-hexachlorocyclohexane by Arthrobacter citreus strain BI-100: identification of metabolites. Journal of General and Applied Microbiology 46(2):59-67.

De Paolis MR, Lippi D, Guerriero E, Polcaro CM, Donati E (2013). Biodegradation of α-, β-, and γ-hexachlorocyclohexane by Arthrobacter fluorescens and Arthrobacter giacomelloi. Applied Biochemistry and Biotechnology 170(3):514-24.

Draft Risk Management Evaluation for Lindane (2007). Persistent Organic Pollutants Review Committee (POPRC) Stockholm Convention on persistent organic pollutants. Retrieved 2018 May 21 from

Egorova DO, Buzmakov SA, Nazarova EA, Andreev DN, Demakov VA, Plotnikova EG (2017). Bioremediation of hexachlorocyclohexane-contaminated soil by the new Rhodococcus wratislaviensis strain Ch628. Water Air Soil Pollution 228(5):183-199.

Endo R, Ohtsubo Y, Tsuda M, Nagata Y (2007). Identification and characterization of genes encoding a putative ABC-type transporter essential for utilization of γ-hexachlorocyclohexane in Sphingobium japonicum UT26. Journal of Bacteriology 189(10):3712-3720.

Fuentes MS, Benimeli CS, Cuozzo SA, Amoroso MJ (2010a). Isolation of pesticide-degrading actinomycetes from a contaminated site: bacterial growth, removal and dechlorination of organochlorine pesticides. International Biodeterioration and Biodegradation 64(6):434-441.

Fuentes MS, Benimeli CS, Cuozzo SA, Saez JM, Amoroso MJ (2010b). Microorganisms capable to degrade organochlorine pesticides. Current Research, Technology and Education Topics in Applied Microbiology and Microbial Biotechnology 1255-1264.

Gerhardt KE, Huang XD, Glick BR, Greenberg BM (2009). Phytoremediation and rhizoremediation of organic soil contaminants: potential and challenges. Plant Science 176(1):20-30.

Giri K, Rawat AP, Rawat M, Rai JPN (2014). Biodegradation of hexachlorocyclohexane by two species of bacillus isolated from contaminated soil. Chemistry and Ecology 30(2):97-109.

Iwasaki I, Utsumi S, Ozawa T (1952). New colorimetric determination of chloride using mercuric thiocyanate and ferric ion. Bulletin of the Chemical Society of Japan 25(3):226-226.

Jennings AA, Li Z (2014). Scope of the worldwide effort to regulate pesticide contamination in surface soils. Journal of Environmental Management 146:420-443.

Kaur J, Moskalikova H, Niharika N, Sedlackova M, Hampl A, Damborsky J, Prokop Z, Lal R (2013). Sphingobium baderi sp. nov., isolated from a hexachlorocyclohexane dump site. International Journal of Systematic and Evolutionary Microbiology 63(2):673-678.

Khan S, Afzal M, Iqbal S, Khan QM (2013). Plant–bacteria partnerships for the remediation of hydrocarbon contaminated soils. Chemosphere 90(4):1317-1332.

Kumar N, Kumar RN, Bora A, Amb MK (2011). An evaluation of pesticide stress induced proteins in three cyanobacterial species Anabaena fertilissima, Aulosira fertilissima and Westiellopsis prolifica using SDS-PAGE. World Academy of Science, Engineering and Technology 5(3):139-146.

Lal R, Pandey G, Sharma P, Kumari K, Malhotra S, et al., Oakeshott JG (2010). Biochemistry of microbial degradation of hexachlorocyclohexane and prospects for bioremediation. Microbiology and Molecular Biology Reviews 74(1):58-80.

Liang XW, Zhang L, Natarajan SK, Beckker DF (2013). Proline Mechanisms of stress survival. Antioxidants & Redox Signaling 19(9):998-1011.

Miguel AS, Roy J, Gury J, Monier A, Coissac E, Ravanel P, Geremia RA, Raveton M (2014). Effects of organochlorines on microbial diversity and community structure in Phragmites australis rhizosphere. Applied Microbiology and Biotechnology 98(9):4257-66.

Moses S, Sinner T, Zaprasis A, Stöveken N, Hoffmann T, Belitsky BR, Sonenshein AL, Bremer E (2012). Proline utilization by Bacillus subtilis: uptake and catabolism. Journal of Bacteriology 194(4):745-758.

Paul S, Paul B, Khan MA, Aggarwal C, Thakur JK, Rathi MS (2013). Effects of Lindane on Lindane-Degrading Azotobacter chroococcum; Evaluation of Toxicity of Possible Degradation Product(s) on Plant and Insect. Bulletin of Environmental Contamination and Toxicology 90(3):351-356.

Pérez DJ, Menone ML, Camadro EL, Moreno VJ (2008). Genotoxicity evaluation of insecticide endosulfan in the wetland macrophyte Bidens laevis L. Environmental Pollution 153(3):695-698.

Perez-Arellano I, Carmona-Alvarez F, Martinez AI, Rodriguez-Diaz J, Cervera J (2010). Pyrroline-5-carboxylate synthase and proline biosynthesis: from osmotolerance to rare metabolic disease. Protein Science 19(3):372-382.

Pesce SF, Wunderlin DA (2004). Degradation of Lindane by a native bacteria consortium isolated from contaminated river sediment. International Biodeterioration and Biodegradation 54(4):255-260.

Philip C, Shrivastava AK, Shrivastava R (2014). Effect of organochlorine pesticide on growth of Actinomycetes. Indian Journal of Scientific Research 4(1):191-195.

Phillips TM, Seech AG, Lee H, Trevors JT (2001). Colorimetric assay for Lindane dechlorination by bacteria. Journal of Microbiological Methods 47(2):181-8.

Polti MA, Aparicio JD, Benimeli CS, Amoroso MJ (2014). Simultaneous bioremediation of Cr (VI) and lindane in soil by Actinobacteria. International Biodeterioration and Biodegradation 88:48-55.

Rajendran UM, Elango K, Anand N (2007). Effects of a Fungicide, an Insecticide and a Biopesticide on Tolypothrix scytonemoides. Pesticide Biochemistry and Physiology 87(2):164-171.

Saez JM, Aparicio JD, Amoroso MJ, Benimeli CS (2015). Effect of the acclimation of a Streptomyces consortium on lindane biodegradation by free and immobilized cells. Process Biochemistry 50(11):1923-1933.

Sagar V, Singh DP (2011). Biodegradation of lindane pesticide by non white-rots soil fungus Fusarium sp. World Journal of Microbiology & Biotechnology 27(8):1747-1754.

Salam JA, Lakshmi V, Das D, Das N (2013). Biodegradation of lindane using a novel yeast strain, Rhodotorula sp. VITJzN03 isolated from agricultural soil. World Journal of Microbiology & Biotechnology 29(3):475-487.

Segura A, Godoy P, van Dillewijn P, Hurtado A, Arroyo N, Santacruz S, Ramos JL (2005). Proteomic analysis reveals the participation of energy- and stress-related proteins in the response of Pseudomonas putida DOT-T1E to toluene. Journal of Bacteriology 187(17):5937- 5945.

Servet C, Ghelis T, Richard L, Zilberstein A, Savoure A (2012). Proline dehydrogenase: a key enzyme in controlling cellular homeostasis. Frontiers in Bioscience 17(1):607-620.

Singh M, Singh DK (2014). Endosulfan induced alteration in bacterial protein profile and RNA yield of Klebsiella sp. M3, Achromobacter sp. M6, and Rhodococcus sp. M2. Journal of Hazardous Materials 265:233-241.

Singh T, Singh DK (2017). Phytoremediation of organochlorine pesticides: Concept, method, and recent developments. International Journal of Phytoremediation 19(9):834-843.

Strungaru SA, Nicoara M, Jitar O, Plavan G (2015). Influence of urban activity in modifying water parameters, concentration and uptake of heavy metals in Typha latifolia L. into a river that crosses an industrial city. Journal of Environmental Health Science & Engineering 13(1):5.

Sukumaran D (2013). Phytoremediation of heavy metals from industrial effluent using constructed wetland technology. Applied Ecology and Environmental Sciences 1(5):92-97.

Vijgen J, Abhilash PC, Li YF, Lal R, Forter M, Torres J, Singh N, Yunus M, Tian C, Schäffer A, Weber R (2011). Hexachlorocyclohexane (HCH) as new Stockholm convention POPs-a global perspective on the management of Lindane and its waste isomers. Environmental Science and Pollution Research 18(2):152-162.

Zaprasis A, Brill J, Thüring M, Wünsche G, Heun M, et al., Bremer E (2013). Osmoprotection of Bacillus subtilis through import and proteolysis of proline-containing peptides. Applied and Environmental Microbiology 79(2):576-587



Under the aegis of Horticulture and Forestry Society from Transylvania