ABSTRACT
Rhamnolipid biosurfactant produced by Pseudomonas aeruginosa, possesses non-toxicity, environmental compatibility, a wide range of pH (4-8), temperature (4-100 °C), and salinity (1-10%) stability. The application of RLs is worldwide accepted in the pharmaceutical, medicinal, and food industries. It has been used for cytotoxicity efficacy analysis with a limited number of cancerous cell lines. To widen the scope of rhamnolipid application as an anticancer agent, we have studied Di-RLs homolog, 'Rha-Rha-C10-C10' produced by Pseudomonas aeruginosa RA5 against human cancerous cell lines including breast cancer (MCF-7), leukemia (K-562), cervical cancer (HeLa), Lung cancer (HOP-62), and colon cancer (HT-29) in a dose-dependent way. It was purified with silica gel chromatography followed by TLC and mass spectroscopy prior to cytotoxicity analysis. With a tensiometer, critical micelle concentration of Di-RLs was estimated to be 33.92 ± 2 mN/m at 0.2%. Cytotoxicity analysis of Di-RLs on K-562 cell line demonstrated inhibition with GI50 and TGI at < 10 µg/mL and 66.6 µg/mL, after 48 h of application. The morphology of human cancerous cell lines was observed under a laser confocal microscope with the SRB staining method. Further research is recommended to comprehend the Di-RLs as a potential anti-cancer agent.
ABSTRACT
High-throughput screening (HTS) is a present-day approach for assaying thousands of cultures in parallel. This miniaturization allows rapid screening of large number of microorganims capable of producing bio-based materials thereby meeting the demands of the ever evolving food, pharmaceutical and cosmetic industry. In this study, agar-based assays for phosphate solubilization, cellulose degradation and lactic acid production were developed in 96-well microplates using Biomek FXP Automated Liquid Handling system. Techno-economic analysis from this study reveals the lower overall cost per assay using HTS as compared to conventional Petri plate assays. Though automated liquid handling workstations have been used to perform liquid-based assays, there are very few studies which report their use for agar-based microplate assays. These findings thus corroborate the establishment of rapid and efficient miniaturized, qualitative agar-based screening methods for identifying microorganisms with potential for commercial application.
