Abstract
Bacterial production of polyunsaturated fatty acids (PUFAs) is a potential biotechnological approach for production of valuable nutraceuticals. Reliable method for screening of number of strains within short period of time is great need. Here, we report a novel simplified method for screening and isolation of PUFA-producing bacteria by direct visualization using the H2O2-plate assay. The oxidative stability of PUFAs in growing bacteria towards added H2O2 is a distinguishing characteristic between the PUFAs producers (no zone of inhibition) and non-PUFAs producers (zone of inhibition) by direct visualization. The confirmation of assay results was performed by injecting fatty acid methyl esters (FAMEs) produced by selected marine bacteria to Gas Chromatography-Mass Spectrometry (GCMS). To date, this assay is the most effective, inexpensive, and specific method for bacteria producing PUFAs and shows drastically reduction in the number of samples thus saves the time, effort, and cost of screening and isolating strains of bacterial PUFAs producers.
Highlights
Microbial lipids are a diverse group of compounds with a number of vital nutraceutical and pharmaceutical applications and utilized commercially since the 1980s
In H2O2-plate assay method, the cells which are susceptible to externally-added H2O2 cannot able to grow suitably and shows a zone of inhibition which is dependant on added concentration of H2O2 on filter paper disc
The contradictory situation was observed for bacterial cells which produce polyunsaturated fatty acids (PUFAs)
Summary
Microbial lipids are a diverse group of compounds with a number of vital nutraceutical and pharmaceutical applications and utilized commercially since the 1980s These microbial lipids or polyunsaturated fatty acids (PUFAs) are obtained from various sources. Selective procedure would allow the detection and isolation of microorganisms producing the desired metabolite This primary screening should be rapid, inexpensive, predictive, specific, but effective over a broad range and should be applicable to large scale. Sometimes primary screening is time consuming and labour intensive when a large number of isolates have to be screened to identify a few potential ones This is possibly the most critical step since it eliminates the large bulk of unwanted isolates, which are either nonproducers or producers of known compounds. This study was based on application of antioxidative effect of PUFA against ROS (Figure 1), for rapid screening of large number of marine isolates. We have presented qualitative method for rapid screening of PUFA producers
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