Chloroplast Isolation from Marine Microalgae

Chloroplasts are an important part of microalgae, macroalgae and plants. As the site of photosynthesis, they are essential for the survival of the organism and are responsible for carbohydrate metabolism, amino acid synthesis, chlorophyll synthesis and lipid synthesis. Marine microalgae is an important dietary microalgae with high nutritional value. Unlike other unicellular microalgae, its cells contain two chloroplasts. In addition, they contain many bioactive compounds that are commercially available. Marine microalgae in particular have unique biochemical properties that higher plants do not have. As an aquatic photosynthetic microorganism, it is able to convert carbon dioxide into biochemical substances, which can then be processed into biofuels, food, feed and high-value bioactive compounds. Sophisticated methods can be used to extract intracellular products, mainly lipids and carotenoids, from microalgae by various processes such as solvent/microwave-assisted extraction, sonication, bead beating, and French filter presses. However, these methods are not suitable for complete organelle isolation as these methods are usually applied to dried biomass, resulting in complete destruction of cells. On the premise of keeping chloroplasts and other organelles intact, efficient cell disruption is a prerequisite for further understanding of various photosynthesis-related phenomena.

Fig. 1. Flow chart depicting scheme for isolation, purification, and characterization of intact chloroplast from microalgal species. (Patil S, et al., 2020)

Services

At present, the chloroplast neutral lipid metabolism of C. reinhardtii has been extensively studied. However, studies on the lipid composition of microalgal chloroplasts have not been comprehensive. A major obstacle to studying the lipid composition of microalgal chloroplasts is the technical challenges associated with isolating and purifying intact chloroplasts. For many years, Lifeasible has been working on the independent isolation of chloroplasts from different plant tissues. Traditionally, Chloroplasts isolated by physically or chemically breaking cell walls and plasma membranes. Here, our scientists developed a method to rapidly isolate chloroplasts from cell-wall-free microalgae. This method is widely applicable to different genera of algae such as Chlamydomonas, Chlorella, and Paracoccus, which has important implications for the determination of antenna size, the understanding of the structure-function relationship of thylakoid membrane components, the sequencing of chloroplast genomes, and the ipid composition of algal organelles. The basic process of marine microalgae chloroplast isolation is as follows:

(1) Microalgal culture and sample collection.

(2) Isolation and purification of intact chloroplast.

(3) The isolated chloroplasts and their integrity were analyzed by phase contrast fluorescence microscopy (Leica) and transmission electron microscopy (TEM).

(4) Enzyme-linked immunosorbent assay (ELISA) to analyze its purity.

(5) The lipid composition of microalgal chloroplasts was identified by ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS/MS).

Competitive Advantages

• A simple, fast, and versatile method to isolate chloroplasts from various microalgae.
• Rapid chloroplast isolation using a two-step method of hypotonic buffer and homogenization saves time and increases yield.
• Chloroplasts purified by this method are mostly intact and contain only minimal contamination.
• It can be used for the study of many different aspects of chloroplast biology and provides a useful reference for the isolation of chloroplasts from other marine microalgae.

Lifeasible can meet the needs of customers on time and on budget through a wide range of chloroplast isolation strategies. Our aim is to be customer-centric and to provide the highest quality service to customers around the world. Our skilled and dedicated scientific researchers ensure that the most appropriate methods and techniques are selected for each specialized chloroplast project. Our customer service representatives are enthusiastic and trustworthy 24 hours a day, 7 days a week. If you are interested in our services, please feel free to contact us for more information or a detailed discussion.

References

1. Patil S, Lali A M, Prakash G. (2020) An efficient algae cell wall disruption methodology for recovery of intact chloroplasts from microalgae[J]. Journal of Applied Biology and Biotechnology. 8(3): 2-8.
2. Zhong Y, Li Y, Xu J, et al. (2020) Isolation of chloroplasts from marine microalgae Isochrysis galbana Parke suitable for organelle lipid composition analysis[J].