Bushra Uzair1, Farid Menaa2, Barkat Ali Khan3, Faryal Vali Mohammad4, Viqar Uddin Ahmad4, Ryad Djeribi5, Bouzid Menaa6. 1. Department of Bioinformatics and Biotechnology, International Islamic University, Islamabad, Pakistan. Electronic address: bushra.uzair@iiu.edu.pk. 2. Department of Advanced Technologies, California Innovations Corporation, San Diego, CA, United States. Electronic address: dr.fmenaa@gmail.com. 3. Faculty of Pharmacy and Alternative Medicine, The Islamia University of Bahawalpur, Pakistan. 4. H.E.J. Research Institute of Biological and Chemical Science, Karachi, Pakistan. 5. Biofilms and Biocontamination of Materials, Annaba University, Algeria. 6. Department of Advanced Technologies, California Innovations Corporation, San Diego, CA, United States.
Abstract
AIMS: Screening of seaweed-associated bacteria capable of producing antimicrobials. METHODS AND RESULTS: Fifteen microbial strains, associated to the brown seaweed Pelvetia canaliculata (Linnaeus) attached to the rocks of Sonmiani Beach (Karachi, Pakistan), were screened. Crude extract filtrates of CMG S2 strain grew on Zobell marine agar (ZMA) had the most remarkable antimicrobial activity. Based on its phenotypic aspects (e.g. Gram-positive, microccoid form), biochemical characteristics (e.g. halotolerance) and genetic analyses, CMG S2 is identified as a putatively new Kocuria marina type strain belonging to the actinobacteria's class and micrococcaceae family. Thereby, the nucleotide sequence analysis of its full-length 16S ribosomal ribonucleic acid (rRNA) gene (GenBank accession number EU073966.1) displayed highest identity (i.e. 99%) and score (2630) with K. marina KMM 3905. Phylogenic trees analysis using the neighbor-joining method showed closest evolutionary distance of CMG S2 with KMM 3905 strain and K. carniphila (DC2201) specie. Interestingly, a unique ultraviolet (UV)-bioactive compound was purified from CMG S2 crude extracts by flash silica gel column and thin-layer chromatography (TLC) techniques. Its chemical structure was unraveled as 4-[(Z)-2 phenyl ethenyl] benzoic acid (PEBA, later named kocumarin) by nuclear magnetic resonance (NMR) spectroscopy techniques. Importantly, kocumarin demonstrated prominent and rapid growth inhibition against all tested fungi and pathogenic bacteria including methicillin-resistant Staphylococcus aureus (MRSA), with a minimal fungal inhibitory concentration (MFC) of 15-25μg/mL and a minimal (bacterial) inhibitory concentration (MIC) of 10-15μg/mL. SIGNIFICANCE AND IMPACT OF THE STUDY: Kocumarin represents a new promising natural antibiotic for in vivo and environmental applications.
AIMS: Screening of seaweed-associated bacteria capable of producing antimicrobials. METHODS AND RESULTS: Fifteen microbial strains, associated to the brown seaweed Pelvetia canaliculata (Linnaeus) attached to the rocks of Sonmiani Beach (Karachi, Pakistan), were screened. Crude extract filtrates of CMG S2 strain grew on Zobell marine agar (ZMA) had the most remarkable antimicrobial activity. Based on its phenotypic aspects (e.g. Gram-positive, microccoid form), biochemical characteristics (e.g. halotolerance) and genetic analyses, CMG S2 is identified as a putatively new Kocuria marina type strain belonging to the actinobacteria's class and micrococcaceae family. Thereby, the nucleotide sequence analysis of its full-length 16S ribosomal ribonucleic acid (rRNA) gene (GenBank accession number EU073966.1) displayed highest identity (i.e. 99%) and score (2630) with K. marina KMM 3905. Phylogenic trees analysis using the neighbor-joining method showed closest evolutionary distance of CMG S2 with KMM 3905 strain and K. carniphila (DC2201) specie. Interestingly, a unique ultraviolet (UV)-bioactive compound was purified from CMG S2 crude extracts by flash silica gel column and thin-layer chromatography (TLC) techniques. Its chemical structure was unraveled as 4-[(Z)-2 phenyl ethenyl] benzoic acid (PEBA, later named kocumarin) by nuclear magnetic resonance (NMR) spectroscopy techniques. Importantly, kocumarin demonstrated prominent and rapid growth inhibition against all tested fungi and pathogenic bacteria including methicillin-resistant Staphylococcus aureus (MRSA), with a minimal fungal inhibitory concentration (MFC) of 15-25μg/mL and a minimal (bacterial) inhibitory concentration (MIC) of 10-15μg/mL. SIGNIFICANCE AND IMPACT OF THE STUDY: Kocumarin represents a new promising natural antibiotic for in vivo and environmental applications.