An efficient algae cell wall disruption methodology for recovery of intact chloroplasts from microalgae

ABSTRACT

The microalgal cell wall breakage has been identified as complex phenomenon which is highly dependent onthe nature and composition of cell wall. A detailed analysis of plastids and their function requires the breakingopen of cell without any damage to cellular components. To develop a rapid and universal methodology forcell wall breakage, liquid nitrogen crushing, sonication, enzymatic lysis, and homogenization procedures wereapplied to various microalgal species. Homogenization-based procedure for the isolation of intact chloroplastwas found to be universal for all algal species under the study. The isolated chloroplasts were subjected tochloroplast integrity analysis. The intact chloroplast exhibited a positive maximum quantum yield and Fv/Fm values ranging from 0.1 to 0.4 as measured by pulse amplitude modulation fluorometry and was found tobe suitable for further downstream applications such as isolation of protein–pigment complexes involved inphotosynthetic O2 evolution. The developed methodology is a quick and efficient technique for the isolation ofintact chloroplasts across different genera of microalgae by employing minor changes in the base protocol as aspecies-specific characteristic