This study's aim was to determine the identity of antibacterial compounds produced by Pseudomonas aeruginosa strain UICC B-40 and describe the antibacterial compounds' mechanisms of action for damaging pathogenic bacteria cells. Isolation and identification of the compounds were carried out using thin layer chromatography (TLC), nuclear magnetic resonance (NMR) spectroscopy and liquid chromatography mass spectrometry (LC-MS) analyses. Antibacterial activity was assayed via minimum inhibitory concentration (MIC) and the antibacterial compound mechanism was observed morphologically through scanning electron microscopy (SEM). This study successfully identified the (2E,5E)-phenyltetradeca-2,5-dienoate antibacterial compound (molecular weight 300 g/mol), composed of a phenolic ester, fatty acid and long chain of aliphatic group structures. MIC values for this compound were determined at 62.5 μg/ml against Staphylococcus aureus strain ATCC 25923. The mechanism of the compound involved breaking down the bacterial cell walls through the lysis process. The (2E,5E)-phenyltetradeca-2,5-dienoate compound exhibited inhibitory activity on the growth of Gram-positive bacteria.
Pseudomonas azotoformans known for the benefit on agriculture area as soil fertility enhancer. Pseudomonas azotoformans UICC B-91 was successfully isolated from tissues of N. altissima which traditionally known for treating infectious diseases. Therefore, endophytic P. azotoformans might contain antimicrobial compound. This research aims on exploring the antimicrobial activities of several extracts P. azotoformans UICC B-91 against pathogenic microbes. Isolates were extracted with dichloromethane, chloroform, and ethyl acetate. Each crude extract was observed for its potential antimicrobial activity with disc diffusion method against several pathogen microbes. Bioactive compound was assayed by thin layer chromatography (TLC) and screened by bioautography. Results indicated that all crude extract have inhibitory effect on Escherichia coli ATCC 8739, Bacillus cereus ATCC 10876, Staphylococcus aureus ATCC 6583, Staphylococcus aureus ATCC 25923, Salmonella typhimurium ATCC 25241, Pseudomonas aeruginosa ATCC 15442, Bacillus subtilis ATCC 19659 and Candida albicans ATCC 10231. Highest clear zone was on dichloromethane extract. TLC fraction of dichloromethane extract was able to inhibit growth on pathogenic bacteria (except for B. subtilis ATCC 19659 and S. aureus ATCC 6583) and fungi at minimum concentration in the range 125-1,000 μg/mL and 125 μg/mL, respectively. Bioactive compound from endophytic P. azotoformans extracted with dichloromethane have antibacterial and anticandidal effect.
Antimicrobial activity analysis were regarding to disc diffusion. 10,11 The antimicrobial activity was measured by diameter of clear zone minus diameter of disc diffusion. Thin Layer Chromatography (TLC)TLC analytical, TLC-bioautography and semipreparative separation were performed on aluminium silica TLC F254 plates. The TLC plates were developed with mobile phase chloroform: methanol (9:1) (F1, F3, F4, and F5) and hexane: ethyl acetate (7:3) (F2). Screening for antimicrobial compound conducted by TLC-bioautography. 12 Positive result reflected on clear zone developed by Rf. Further TLC-separation on clear zone-Rf were developed with chloroform: ethyl acetate (5:5) (F1, F4, and F5).
The infection disease is still important problem in Indonesia, especially in eastern part of Indonesia. The problem of the bacterial resistance has resulted in increased morbidity, mortality, and costs of healthcare. This situation is therefore drives many researches on discovery for new antibiotics and novel bioactive compounds to combat multidrug resistant microbes, such as Mycobacterium tuberculosis, Staphylococcus spp., and Streptococcus spp. The aim of this study is to know the development of the endophytic microorganisms. The endophytic bacteria provide vast potential in producing various novel natural products, including secondary metabolites similar to their hosts. The plant is one of host for the endophytic microorganisms reside. In Indonesia, that endophytic is used medicinally for treatment as well as from the host. The bioactive compound of endophytic microorganisms are used medicinally for treatment antibacterial, antiviral, anticancer, antioxidants, antidiabetic, and immunosuppressive. These showed that endophytic bacteria are promising source of new antibiotics against susceptible and resistant forms of various infectious microbes.
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