Engineering bacteria for bioremediation of persistent organochlorine pesticide lindane (γ-hexachlorocyclohexane)

• Published in: Bioresource technology Vol. 149; pp. 439 - 445
• Main Authors: Chaurasia, Akhilesh Kumar, Adhya, Tapan Kumar, Apte, Shree Kumar
• Format: Journal Article
• Language: English
• Published: Kidlington Elsevier Ltd 01.12.2013; Elsevier
• Subjects: Anabaena; Anabaena - genetics; Anabaena - metabolism; Bacteria; Bacterial Proteins - metabolism; Biodegradation, Environmental; Bioengineering; Biological and medical sciences; Bioremediation; Biotechnology; Degradation; Environment and pollution; Escherichia coli - metabolism; Fundamental and applied biological sciences. Psychology; Genes, Bacterial; Genetic Engineering; Industrial applications and implications. Economical aspects; LinA2 overexpression; Lindane - metabolism; Lindane bioremediation; Lindane toxicity and detection; Pesticides; Pesticides - metabolism; Recombinant; Recombinant E. coli/Anabaena strains; Recombination, Genetic - genetics; Sphingomonas; Stress, Physiological; Toxicity; Xenobiotics; LinA2 overexpression; Lindane toxicity and detection; Recombinant E. coli/Anabaena strains; Lindane bioremediation; Recombination; Insecticide; Toxicity; Lindane; Pollution control; Pesticides; Gene overexpression; Engineering; Organochlorine compounds; Cyanobacteria; Anabaena; Bacteria; Bioremediation
• ISSN: 0960-8524; 1873-2976
• DOI: 10.1016/j.biortech.2013.09.084

•GroEL expression identified as a major target for lindane toxicity in Anabaena.•Recombinant Anabaena strain developed to bioremediate lindane in paddy fields.•LinA2 overexpression facilitated degradation of all isomers of lindane by GM Escherichia coli.•Fluorescence quenching-based method designed for visual lindane detection on plates.•Degradation of high lindane levels (2mg/ml) by dead bioengineered E. coli demonstrated. Strategies were designed for bioremediation of the highly persistent toxic pesticide γ-hexachlorocyclohexane (γ-HCH) or lindane from the environment. Lindane caused the loss of stress-protective chaperone GroEL, and inhibited photosynthesis, respiration and nitrogen-fixation in Anabaena, resulting in growth arrest. To alleviate lindane toxicity, the linA2 gene, encoding HCH dehydrochlorinase from Sphingomonas paucimobilis B90, was knocked-in at an innocuous locus in Anabaena genome and over-expressed from an eco-friendly light-inducible PpsbA1 promoter. The recombinant Anabaena degraded >98% of 10ppm lindane within 6–10days. A LinA2 overexpressing Escherichia coli strain could degrade 10ppm of all the isomers of lindane within 1h and displayed a visual degradation zone on a newly designed histochemical plate containing 50mg lindane within 12h. The study demonstrates (a) bioremediation of traces of lindane prevalent in paddy fields, using bioengineered photoautotrophic Anabaena, and, (b) biodegradation of huge stockpiles of lindane, by employing recombinant live/dead E. coli.