Biodecomposition of Phenanthrene and Pyrene by a Genetically Engineered Escherichia coli

Abstract

Background: Genetically engineered microorganisms (GEMs) can be used for bioremediation of the biological pollutants into nonhazardous or less-hazardous substances, at lower cost. Polycyclic aromatic hydrocarbons (PAHs) are one of these contaminants that associated with a risk of human cancer development. Genetically engineered E. coli that encoded catechol 2,3- dioxygenase (C230) was created and investigated its ability to biodecomposition of phenanthrene and pyrene in spiked soil using high-performance liquid chromatography (HPLC) measurement. We revised patents documents relating to the use of GEMs for bioremediation. This approach have already been done in others studies although using other genes codifying for same catechol degradation approach. Objective: In this study, we investigated biodecomposition of phenanthrene and pyrene by a genetically engineered Escherichia coli. Methods: Briefly, following the cloning of C230 gene (nahH) into pUC18 vector and transformation into E. coli Top10F, the complementary tests, including catalase, oxidase and PCR were used as on isolated bacteria from spiked soil. Results: The results of HPLC measurement showed that in spiked soil containing engineered E. coli, biodegradation of phenanthrene and pyrene comparing to autoclaved soil that inoculated by wild type of E. coli and normal soil group with natural microbial flora, were statistically significant (p<0.05). Moreover, catalase test was positive while the oxidase tests were negative. Conclusion: These findings indicated that genetically manipulated E. coli can provide an effective clean-up process on PAH compounds and it is useful for bioremediation of environmental pollution with petrochemical products.

Insights Into Mechanism of the Naphthalene-Enhanced Biodegradation of Phenanthrene by Pseudomonas sp. SL-6 Based on Omics Analysis

“…Phenanthrene was extracted with isochoric n-hexane, then redissolved with methanol after volatilizing n-hexane. Sterilization with 0.22 μm filter, residual PHE was determined by high performance liquid chromatography (HPLC) ( Wang et al, 2015 ; Maryam et al, 2020 ). HPLC conditions: the column was ZORBAX Eclipse XDB-C18 (4.6 × 150 mm, 5 μm), injection volume = 5 μL, column temperature = 24°C, flow rate = 1 mL/min, methanol: water = 70: 30, signal = 254 nm.…”

Alternative Strategies for Microbial Remediation of Pollutants via Synthetic Biology

“…Moreover, environment and ecosystem risk assessments infer unpredictable and adverse effects, as discussed above (Cervelli et al, 2016;van Dorst et al, 2020). Particularly, the ecological risk assessment behind addition of GEMs (Genetically Engineered Microorganisms) (Benjamin et al, 2019;Ahankoub et al, 2020) to the native environment rather than a laboratory (Fernandez et al, 2019) is done mainly because of unnecessary delivery of antibiotic resistance marker along with recombinant genome of interest (Davison, 2002;Nora et al, 2019), and unintentional uptake or transfer of induced genes to other microorganisms (French et al, 2019;Janssen and Stucki, 2020). This phenomenon is definately disturbing the microbial native genome entity (Gangar et al, 2019;Petsas and Vagi, 2019).…”