Recombinant Hydrolase Mediated Biodegradation of Fenamiphos by Cronobacter muytjensii GH10: Gene Cloning, Expression, and Docking Analysis | ||||
| Egyptian Journal of Chemistry | ||||
| Article 49, Volume 68, Issue 12, December 2025, Page 627-640 PDF (6.48 MB) | ||||
| Document Type: Original Article | ||||
| DOI: 10.21608/ejchem.2025.395474.11922 | ||||
 View on SCiNiTO | ||||
| Authors | ||||
Ghada M. El-Sayed 1; nivien Abd-Rahman Abo Serih 2; Amira Mohammed-Roshdy Ibrahim Ibrahim Asar 3; Maher A. Hammad A. Hammad4; Shyamaa H. Mahmoud5; Omaima A Sharaf  6
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| 1Department of Microbial Genetics, National Research Centre | ||||
| 2Microbial Genetic Department, Biotechnology Research Institute, national research centr | ||||
| 3Microbial Chemistry department, Biotechnology institution, National Research Centre, Egypt | ||||
| 4Department of Plant Protection, Faculty of Agriculture, Ain Shams University, Cairo 11566, Egypt | ||||
| 5Zoology Department, Faculty of Science , Menoufia University, Egypt | ||||
| 6Department of Agricultural Microbiology, Biological and Agricultural division, National Research Centre | ||||
| Abstract | ||||
| Abstract The repeated application of pesticide, fenamiphos is a common agricultural practice in greenhouse cultivation areas in Egypt, resulting in the accumulation of significant fenamiphos residues in the soil. The aim of this study is to eliminate the fenamiphos residues from contaminated soil using the recombinant enzyme. For this purpose, a strain of Cronobacter muytjensii GH10 was isolated from soils treated with chlorpyrifos. It demonstrated notable efficiency in degrading fenamiphos in both in vitro assays and pot experiments conducted in sterilized and non-sterilized soils. A novel enzyme capable of hydrolyzing the P–O–C bond of fenamiphos was identified from C. muytjensii GH10, and its gene (oph) was successfully cloned and heterologously expressed in Escherichia coli BL21 (DE3) using a pET expression vector under the control of the T7 promoter system. The recombinant hydrolase encoded by the oph gene exhibited superior biodegradation activity compared to the native strain, particularly in vitro and in pot experiments using sterilized soil. Furthermore, in silico analysis of the predicted protein structure encoded by oph revealed enhanced insights into the enzyme's binding affinity and catalytic function. Notably, two hydrogen bonds were identified between fenamiphos and the hydrolase via the THR183 residue, supporting the enzyme's role in cleaving the P–O–C bond. This study is the first to evaluate both the bacterial cultural of C. muytjensii GH10 and its recombinant hydrolase enzyme for the biodegradation of fenamiphos. | ||||
| Keywords | ||||
| fenamiphos; biodegradation; recombinant hydrolase; gene cloning and expression; molecular docking | ||||
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