A novel method for the release of viable single cells from Botryococcus braunii (Race B) colony using iodine treatment

Abstract

The first report on the use of Iodine solution for single cell release from colonial green algae Botryoccus braunii is described. The possible reaction of Iodine (I2), Potassium iodide (KI) and the combination of both KI + I2 solutions to release single cells embedded in the hydrocarbon-rich extracellular matrix (ECM) of B. braunii have been attempted. After these solution treatments, the algal samples were immediately viewed under a microscope to check the release of single cells and the average colonies released were counted using hemocytometer within a specific time interval. Treated colonies were initially filtered using 5 μm mesh, the remaining filtrate was passed through 0.45 μm membrane filter and the resultant organisms were subjected to various analyses. After treatment, >90% of viable cells were released by Iodine solution, without having any inhibitory effect or damage on the released cells. Compared to the above results, no cells were released by KI and < 2% cells were released and colonies were rapidly stained dark brown by KI + I2 solution. Nile red analysis revealed that the released cells were rich in lipid bodies and the FTIR spectrum of extracted liquid hydrocarbons revealed that the presence of various forms of iodinated hydrocarbons under Iodine treatment. The experiment also proved the mechanism behind cell release by a liquid hydrocarbon, squalene, which reacted with Iodine atoms. Here, Iodine takes part to catalyst in the breakdown of double bonds to form Squalene-HI. The extracted hydrocarbons from Iodine treated shells can be utilized for pharmaceutical and cosmeceutical industries.