Citation: TAO Zhan-Hua, LIU Jun-Xian, SHI De-Qiang, KANG Jian. Analysis of Staphyloxanthin Biosynthesis Using Single Cell Raman Spectroscopy[J]. Chinese Journal of Analytical Chemistry, ;2016, 44(3): 456-461. doi: 10.11895/j.issn.0253-3820.150729
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Laser tweezers Raman spectroscopy (LTRS) was used to study the inhibitory effect of indol on staphyloxanthin biosynthesis in Staphylococcus aureus cells and the dynamic changes of this pigment content inside bacterial cells during batch cultivation. The Raman spectra of Staphylococcus aureus cells cultivated for different time and exposed to various doses of indol were acquired. The intensity of 1523 cm-1 band was used for the quantification of staphyloxanthin, in the meantime, the pigment was measured by UV spectrometry. The experimental result showed that an excellent linear relationship existed between the intensities of Raman peak at 1523 cm-1 of bacterial cells and the pigment contents estimated by UV spectrometry, with a correlation coefficient of 0.9772. The spectral data at population level as well as single cell level revealed that indol could inhibit the production of pigment in dose-dependent manner, and the pigment content in bacterial cells incubated with indol decreased by 70%. Under the batch growth condition, the pigment amount in Staphylococcus aureus cells reached the maximum value during the middle exponential growth phase (12 h) and the heterogeneity of pigment content in bacterial cells within certain populations at various time points was relatively small, with RSDs of 39.2% to 61.1%. This investigation indicates that LTRS can be served as a reliable method for the analysis of staphyloxanthin content at single cell level.
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