Fingerprinting analysis of photopigments in purple bacteria

Abstract:

Abstract:［Objective］Photopigments，including carotenoid and bacteriochlorophyll a，are the most important functional units of photosynthesis in purple bacteria． We developed rapid qualitative and quantitative methods to determine photopigments．［Methods］ Using Rhodopesudomonas palustris CQV97 as a reference，we used image gray intensity analysis，absorption spectrophotometry，thin layer chromatography (TLC)，HPLC and mass spectrometry (MS) for photopigment analysis．［Results］ The total amount of photopigments increased by 13.5% by using modified acetonemethanol extraction． We developed two types of photopigment fingerprintings by TLC and HPLC，estimated the apparent relative content of each photopigment of fingerprintings，and determined the corresponding relationships between Rf value of each photopigment on TLC fingerprinting and retention time of each photopigment elution in HPLC fingerprinting． Based on the data from the absorption spectra，MS and related photopigment biosynthetic pathway analysis，we identified 11 photopigments in CQV97 strain． Using this strain as a standard，we analyzed photopigments of the tested samples by TLC or HPLC． It was shown that (1) the relative standard deviation (RSD) of the two methods was less than 5%; (2) the compositions and contents of the theory sample were consistent with that of the standard sample; (3) the photopigment compositions of the real sample was the same as the standard sample，but the photopigment content was different． ［Conclusion］ Both of TLC and HPLC analyses for photopigment determination have good stability and repeatability． The fingerprintings analyses are suitable for rapid determination of photopigments of purple bacteria and have important application in control of regulation mechanism for photopigment synthesis．