Molecular Characterization of spa, hld, fmhA, and l ukD Genes and Computational Modeling the Multidrug Resistance of Staphylococcus Species through Callindra harrisii Silver Nanoparticles

Muhammad Waseem; Muhammad Naveed; Shafiq Ur Rehman; Syeda Izma Makhdoom; Tariq Aziz; Metab Alharbi; Abdulrahman Alsahammari; Abdullah F Alasmari

doi:10.1021/acsomega.3c01597

Molecular Characterization of spa, hld, fmhA, and l ukD Genes and Computational Modeling the Multidrug Resistance of Staphylococcus Species through Callindra harrisii Silver Nanoparticles

ACS Omega. 2023 Jun 5;8(23):20920-20936. doi: 10.1021/acsomega.3c01597. eCollection 2023 Jun 13.

Authors

Muhammad Waseem¹, Muhammad Naveed¹, Shafiq Ur Rehman², Syeda Izma Makhdoom¹, Tariq Aziz³, Metab Alharbi⁴, Abdulrahman Alsahammari⁴, Abdullah F Alasmari⁴

Affiliations

¹ Department of Biotechnology, Faculty of Science and Technology, University of Central Punjab, Lahore 54590, Pakistan.
² Department of Basic and Applied Chemistry, Faculty of Science and Technology, University of Central Punjab, Lahore 54000, Pakistan.
³ Department of Agriculture, University of Ioannina, Arta 47100, Greece.
⁴ Department of Pharmacology and Toxicology, College of Pharmacy, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia.

Abstract

The problem of multidrug resistance in bacterial pathogens is significant and is related to the high morbidity and death rates of living things due to increased levels of beta-lactamases. Plant-derived nanoparticles have gained a great significance in the field of science and technology to combat bacterial diseases, especially multidrug-resistant bacteria. This study examines the multidrug resistance and virulent genes of identified pathogenic Staphylococcus species obtained from Molecular Biotechnology and Bioinformatics Laboratory (MBBL), culture collection. The polymerase chain reaction-based characterization of Staphylococcus aureus and Staphylococcus argenteus having ON875315.1 and ON876003.1 accession IDs revealed the presence of the spa, LukD, fmhA, and hld genes. The green synthesis of silver nanoparticles (AgNPs) was carried out by utilizing the leaf extract of Calliandra harrisii, of which metabolites act as capping and reducing agents for the precursor of nano-synthesis, i.e., AgNO₃ of 0.25 M. The synthesized AgNPs were characterized via UV-vis spectroscopy, Fourier transform infrared spectroscopy, scanning electron microscopy, and energy-dispersive X-ray analysis which inferred the bead-like shape of our nanoparticles with the size of 2.21 nm with the existence of aromatic and hydroxyl functional groups at surface plasmon resonance of 477 nm. The antimicrobial activity by AgNPs showed 20 mm inhibition of Staphylococcus species as compared to the vancomycin and cefoxitin antibiotics along with crude plant extract, which showed a minimum zone of inhibition. The synthesized AgNPs were also analyzed for various biological activities like anti-inflammatory with 99.15% inhibition in protein denaturation, antioxidant with 99.8% inhibition in free radical scavenging, antidiabetic with 90.56% inhibition of alpha amylase assay, and anti-haemolytic with 89.9% inhibition in cell lysis which shows good bioavailability and biocompatibility of the nanoparticles with the biological system of the living being. The amplified genes (spa, LukD, fmhA, and hld) were also analyzed for their interaction with AgNPs computationally at the molecular level. The 3-D structure of AgNP and amplified genes was retrieved from ChemSpider (ID: 22394) and Phyre2 online server, respectively. The binding affinities of AgNP with spa, LukD, fmhA, and hld were -7.16, -6.5, -6.45, and -3.3 kJ/mol, respectively, which infers a good docking score except of hld which is -3.3 kJ/mol due to its small size. The salient features of biosynthesized AgNPs proved to be an effective approach in combating the multidrug-resistant Staphylococcus species in the future.