Characterization of water-soluble dark-brown pigment from Antarctic bacterium, Lysobacter oligotrophicus
Section snippets
Extraction and purification of the pigment
L. oligotrophicus strain 107-E2T was cultivated on a rotary shaker in 0.25xLB medium (2.5 g/L of tryptone, 1.3 g/L of yeast extract) at 23°C for 2 weeks (25). The culture containing dark-brown pigment was centrifuged (5000 ×g, 10 min) and the supernatant was filtrated by 0.22 μm pore size filter (Millipore). Then flow-through fraction was acidified to pH 2.5 with HCl and incubated on overnight. Precipitate was collected by centrifuge (5000 ×g, 5 min), washed by ethanol twice, and dried. For
Purification and identification of the pigment
In the previous study, the pigment from L. oligotrophicus was expected to be melanin pigment (25). The dark-brown water-soluble pigment from L. oligotrophicus was recovered from the culture by the modified method for bacterial melanin isolation (17). To identify the pigment, spectral (UV–visible, EPR and FT-IR) analyses were performed (Fig. 1, Fig. 2, Fig. 3). When UV–visible absorption spectra of the pigment at 180–750 nm were analyzed, absorption was observed at the all observed wavelengths (
Acknowledgments
The authors would like to express their sincere thanks to all members concerned to the 46th Japanese Antarctic Research Expedition. This study was supported by JSPS KAKENHI (19205022 and 23657068).
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