Recombinant Hydrolase Mediated Biodegradation of Fenamiphos by Cronobacter muytjensii GH10: Gene Cloning, Expression, and Docking Analysis

El-Sayed, Ghada M.; Abo Serih, Nivien Abd-Rahman; Asar, Amira Mohammed-Roshdy Ibrahim Ibrahim; Hammad, Maher A. Hammad A.; Mahmoud, Shyamaa H.; Sharaf, Omaima A

doi:10.21608/ejchem.2025.395474.11922

Recombinant Hydrolase Mediated Biodegradation of Fenamiphos by Cronobacter muytjensii GH10: Gene Cloning, Expression, and Docking Analysis

Document Type : Original Article

Authors

¹ Department of Microbial Genetics, National Research Centre

² Microbial Genetic Department, Biotechnology Research Institute, national research centr

³ Microbial Chemistry department, Biotechnology institution, National Research Centre, Egypt

⁴ Department of Plant Protection, Faculty of Agriculture, Ain Shams University, Cairo 11566, Egypt

⁵ Zoology Department, Faculty of Science , Menoufia University, Egypt

⁶ Department of Agricultural Microbiology, Biological and Agricultural division, National Research Centre

10.21608/ejchem.2025.395474.11922

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.

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