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Abstract
The phenylurea herbicides are persistent in soil and water, necessitating the creation of methods for removing them from the environment. This study aimed to examine the soil microbial diversity, searching for local bacterial isolates able to efficiently degrade the phenylurea herbicide isoproturon, 3-(4-isopropylphenyl)-1, 1-dimethylurea (IPU). The best isolates able to effectively degrade IPU were selected, characterized, and identified as Pseudomonas putida and Acinetobacter johnsonii. The catechol 1, 2-dioxygenase enzyme's catA gene was amplified, cloned, and expressed in E. coli M15. The Expressed E. coli showed high degradation efficiency (44.80%) as analyzed by HPLC after 15 days of inoculation in comparison to P. putida (21.60%). The expression of the catA gene in P. putida and expressed E. coli was measured using quantitative polymerase chain reaction (qPCR). The results displayed a significant increase in the mRNA levels of the catA gene by increasing the incubation time with IPU. Hydrophilic interaction chromatography (HILIC) mass spectrometry analysis revealed that three intermediate metabolites, 1-(4-isopropylphenyl)-3-methylurea (MDIPU), 4-Isopropylaniline (4-IA) and 1-(4-isopropylphenyl) urea (DDIPU) were generated by both P. putida and expressed E. coli. In addition, IPU-induced catA activity was detected in both P. putida and expressed E. coli. The supernatant of both P. putida and expressed E. coli had a significant influence on weed growth. The study clearly exhibited that P. putida and expressed E. coli were capable of metabolizing IPU influentially and thus could be utilized for bioremediation and biodegradation technology development.
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Details
1 Cairo University, Department of Genetics, Faculty of Agriculture, Giza, Egypt (GRID:grid.7776.1) (ISNI:0000 0004 0639 9286)
2 Cairo University, Department of Biochemistry, Faculty of Agriculture, Giza, Egypt (GRID:grid.7776.1) (ISNI:0000 0004 0639 9286); Egypt-Japan University of Science and Technology (E-JUST), Basic Applied Science institute, New Borg El-Arab City, Egypt (GRID:grid.440864.a) (ISNI:0000 0004 5373 6441)
3 Cairo University, Botany and Microbiology Department, Faculty of Science, Giza, Egypt (GRID:grid.7776.1) (ISNI:0000 0004 0639 9286)
4 Misr University for Science and Technology (MUST), College of Biotechnology, 6(th) October City, Egypt (GRID:grid.440875.a) (ISNI:0000 0004 1765 2064)