ANTIBACTERIAL AND ANTICANCER ACTIVITIES OF MARINE BACTERIAL EXTRACTS AND DETECTION OF GENES FOR BIOACTIVE COMPOUNDS SYNTHESIS
Keywords:
Antibacterial, Metabolite, Sponge-associated bacteria, Anticancer, PKS, NRPSAbstract
Objective: This study was aimed to determine the antimicrobial, toxicity, and anticancer activities of ethyl acetate extracts from sponge-associated bacteria and to detect the presence of PKS and NRPS genes.
Methods: Crude extracts of sponge-associated bacteria HAA-01, HAL-13, and HAL-74 were used as samples for toxicity test using Brine Shrimp Lethality Test (BSLT). The Minimum Inhibitory Concentration (MIC) of bacterial extracts were tested using agar diffusion method. Cytotoxicity test was carried out by 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT)method against HeLa cell line. To determine the presence of keto synthase (KS) domain of polyketide synthase (PKS) and adenylation (A) domain of nonribosomal peptide synthetase (NRPS) genes were carried out using Polymerase Chain Reaction (PCR). DNA of PCR products were purified and sequenced. The DNA sequences were analyzed by the Blast program, and phylogenetic tree analysis was constructed using MEGA5 program with neighbor-joining method.
Results: The research results indicated that the best value of MIC of bacterial extracts against five bacteria such as Escherichia coli, Enteropathogenic E. coli (EPEC), Pseudomonas aeruginosa, Staphylococcus aureus and Bacillus subtilis were 0.1 mg/ml. The half maximal lethal concentration (LC50) value of HAL-74, HAA-01 and HAL-13 bacterial metabolite extracts against the brine shrimp Artemia salina were 378 μg/ml, 463.028 μg/ml and 444.796 μg/ml, respectively. The bacterial metabolite extract has cytotoxicity effect on HeLa cell line with the best half maximal inhibitory concentration (IC50) value was 132.877 µg/ml as performed by extract of HAL-74. Genetic analysis showed that HAL-74 and HAA-01 genomes contained genes that encoded the KS domain of PKS gene and A domain of NRPS gene, while HAL-13 genome encoded the KS domain of PKS gene, which played a role in the synthesis of bioactive compounds.
Conclusion: Bacterial metabolite extracts of HAA-01, HAL-13, and HAL-74 had antimicrobial, toxicity against A. salina, and anticancer against HeLa cell line activities. The best toxicity and anticancer activity were performed by HAL-74 crude extract with LC50 and IC50 were 378 mg/ml and 132.877 mg/ml, respectively. HAA-01 and HAL-74 genomes contained PKS and NRPS genes while HAL-13 genome contained PKS gene.
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References
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