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. 2012:2012:542721.
doi: 10.1155/2012/542721. Epub 2012 Aug 15.

Novel simplified and rapid method for screening and isolation of polyunsaturated Fatty acids producing marine bacteria

Ashwini Tilay  1 , Uday Annapure
Affiliations

Novel simplified and rapid method for screening and isolation of polyunsaturated Fatty acids producing marine bacteria

Ashwini Tilay et al. Biotechnol Res Int. 2012.

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 H(2)O(2)-plate assay. The oxidative stability of PUFAs in growing bacteria towards added H(2)O(2) 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.

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Figures

Figure 1
Figure 1
Proposed mode of action for the H2O2-plate assay method.
Figure 2
Figure 2
Map of western coast region of Maharashtra (INDIA).
Figure 3
Figure 3
Protocol for primary screening of marine isolates.
Figure 4
Figure 4
H2O2-plate assay.
Figure 5
Figure 5
Gas chromatogram showing fatty acid profile of a selected marine isolate after primary screening.
See this image and copyright information in PMC

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