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Selection and Molecular Response of AHL-lactonase (aiiA) Producing Bacillus sp. Under Penicillin G-induced Conditions

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Abstract

Quorum sensing (QS) is the process by which microorganisms employ chemicals called autoinducers (AIs) to communicate with their population. The QS mechanism generally controls the expression of the virulence related genes in bacteria. N-acyl homoserine lactones (AHLs) are the most widespread QS molecules. Due to their diverse AHL-lactonase activities, Bacillus species make particularly suitable candidates for procedures such as demolition of pathogenic bacterial QS signals and bioremediation of β-lactam antibiotics from contaminated environments. In this study, seven Bacillus strains with Quorum quenching (QQ) activity were isolated using an enrichment medium supplemented with Penicillin G (PenG). The AHL-lactonase encoding gene (aiiA) was amplified by PCR and sequenced. Amino acid sequences underwent multiple sequence alignment. Docking studies were carried out with both C6HSL and PenG ligand using AutoDock tools. The aiiA amino acid sequences of the isolates were found to be well conserved. Furthermore, amino acid sequence alignment revealed that 74.9% of amino acid sequences were conserved in the genus Bacillus. Docking of the C6HSL to wild type (3DHA) and H97D variant reduced the docking score by only 0.1 kcal/mol for the mutated protein. When PenG docked with a higher (1.5 kcal/mol) score as a ligand to wild-type and mutant receptors, the docking score for the mutated protein likewise decreased by 0.1 kcal/mol. This research contributed to the diversification of organisms with QQ activity and beta-lactam antibiotic resistance. It also clarified the binding score of the PenG ligand to the Bacillus AHL lactonase molecule for the first time.

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Data Availability

The datasets created and/or analyzed for this report are accessible from the corresponding author on reasonable request.

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Acknowledgements

The authors would like to thank employees of Kılıç Seafood Inc., Didim Incubation and Adaptation Facility, for their support. Professor Dr. Özgür Ceylan’s contribution to this study is also gratefully acknowledged.

Funding

This project was completed with the generous financial support of Ege University under grant number FM-GAP-2022-24187.

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Authors and Affiliations

  1. Graduate School of Natural and Applied Sciences, Ege University, İzmir, Turkey

    Gülperi Bulut

  2. Faculty of Sciences, Department of Biology, Ege University, İzmir, Turkey

    İhsan Yaşa & Asiye Esra Eren Eroğlu

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Gülperi Bulut, İhsan Yaşa and Asiye Esra Eren Eroğlu planned the study. Gülperi Bulut and Asiye Esra Eren Eroğlu carried out laboratory tests, examined the data, and wrote the manuscript. İhsan Yaşa assessed the manuscript for scope. The manuscript has passed review by all authors, and all of them agree to submit for publication.

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Correspondence to Asiye Esra Eren Eroğlu.

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Bulut, G., Yaşa, İ. & Eren Eroğlu, A.E. Selection and Molecular Response of AHL-lactonase (aiiA) Producing Bacillus sp. Under Penicillin G-induced Conditions. Protein J 42, 427–436 (2023). https://doi.org/10.1007/s10930-023-10115-7

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