evaluating chlorophyll content in chlorella sorokiniana and haematococcus pluvialis to study co2, light color, and intensity effects

Enhancing Chlorophyll Production in Microalgae via Growth Factor Optimization

In recent years, the growing environmental problems caused by anthropogenic activities and their adverse effects on the health and balance of ecosystems have driven the search for more efficient and environmentally friendly production systems. This has accelerated processes in industries and fostered the implementation of bioremediation treatments and the development of bio compounds to mitigate these harmful effects1.
This context has led to a significant growth in the study of microalgae, driven by the need to find innovative solutions to current environmental and economic challenges. Microalgae thrive in aquatic environments, using sunlight and carbon dioxide as sources of energy and carbon, respectively. This characteristic makes them a sustainable and promising alternative for producing various valuable products. Research in this field has focused on understanding the physiology and metabolism of these cells, as well as developing efficient technologies for their cultivation and processing2.
There is a clear need to develop accessible and reliable tools for studying microalgae to accelerate research processes and deepen the understanding of their physiology, metabolism, and potential applications. These tools should enable rapid analysis of the effect of environmental and production factors on key parameters, such as chlorophyll concentration, which is a fundamental indicator of the health and development of these organisms. A decrease in chlorophyll content may indicate environmental stress, nutritional deficiencies, or diseases.
These green pigments play a crucial role in photosynthesis, capturing sunlight energy to convert carbon dioxide and water into glucose and oxygen and constitute between 0.5% and 5% of the biomass of microalgae3. Beyond their essential role in sustaining life processes, chlorophylls have found applications in various industries. Chlorophyll extracts are utilized in food and beverage processing as natural colorants, imparting vibrant green hues to products while also providing antioxidant properties. Moreover, chlorophyll-based supplements are gaining popularity in the health and wellness sector due to their purported detoxifying and anti-inflammatory effects. By harnessing the multifaceted properties of chlorophyll, industries can develop innovative products that contribute to both visual appeal and consumer well-being1.
In this regard, one microalgae of great biotechnological relevance is C. sorokiniana. This microorganism stands out for its rapid growth rate, making it highly efficient in biomass production. Additionally, C. sorokiniana exhibits a diverse content of highly nutritional compounds, including proteins, lipids, and vitamins, which renders it valuable for various applications in food, feed, and biofuel production2. Moreover, this microalgae species has been found to produce extracellular enzymes with diverse functions, opening up possibilities for biotechnological applications such as wastewater treatment, bioremediation, and pharmaceuticals4. In addition to its rapid growth and versatile applications, C. sorokiniana also demonstrates significant potential for chlorophyll production. As a photosynthetic microorganism, C. sorokiniana possesses the machinery necessary for the synthesis of chlorophyll, which constitutes between 0.5% and 5% of the biomass of microalgae3. This capability makes C. sorokiniana an attractive candidate for commercial-scale chlorophyll production and promises to be a sustainable solution to urgent environmental and nutritional challenges5.
On the other hand, another microalgae species of significant interest is H. pluvialis. This microalga is renowned for its production of astaxanthin, a potent antioxidant pigment with numerous industrial applications. Astaxanthin serves as a protective mechanism for H. pluvialis photosystem, shielding it from oxidative stress induced by environmental factors. This pigment is highly sought after in cosmetics, nutraceuticals, and aquaculture industries, owing to its antioxidant properties and potential health benefits. With its abundant ability to produce astaxanthin, H. pluvialis presents a promising avenue for developing innovative products catering to various industrial and consumer needs6.
Recent studies have also highlighted H. pluvialis1 potential for chlorophyll production, further enhancing its industrial significance. For instance, a study investigated the chlorophyll content of H. pluvialis under varying growth conditions and found that under optimal cultivation parameters, H. pluvialis exhibited remarkable chlorophyll production rates, surpassing those of other microalgae species7. This finding underscores the potential of H. pluvialis as a sustainable source of chlorophyll, offering novel opportunities for its utilization in various industrial sectors.

Author Spotlight: Optimizing Growth Factors for Production of Biotechnologically Relevant Secondary Metabolites

Evaluation of the Effect of Growth Factors on Chlorophylls a and b Production from Microalgae

Our project evaluates how growth factor control affects the production of biotechnologically-relevant secondary metabolites such as chlorophyll from microalgae such as Chlorella sorokiniana and Haematococcus pluvialis at different scales. We have focused on establishing an analytical method to quantify chlorophylls in microalgae. Some emerging technologies for advancement in this are the modeling and optimization of processes using computational technique.However, in order to carry them out, it is necessary to obtain experimental information on the processes. The growth factors evaluated in Chlorella sorokiniana that have the greatest effect on chlorophyll production are the light color and the carbon dioxide supply flow. The changing light intensity does not significantly influence the production of this metabolite.The findings in this field of study can be advanced by implementing growth factors that induce microalgae to increase the production of secondary metabolites in conjunction with rapid, simple, and reliable analysis techniques. Our work provide those with guidelines to investigate and understand the biochemical and the energetic processes carried out by microalgae with different substrates, evaluating the effect of environmental variables to optimize the production of your mass and cellular compounds for various applications.

Evaluating Chlorophyll Content in Chlorella sorokiniana and Haematococcus pluvialis to Study CO2, Light Color, and Intensity Effects

To begin, prepare the inoculum to study the growth kinetics of Chlorella sorokiniana and Haematococcus pluvialis Prepare the required volume of sterile culture medium according to standard laboratory procedures. Configure the light source for both the photobioreactor and the flask to the desired color spectrum. Cover both systems to prevent external light interference.After adjusting the photoperiod settings, connect the carbon dioxide duration system to the photobioreactor to provide carbon dioxide for 12 hours to chlorella. Next, inoculate the culture vessel to achieve a starting cell density of 300, 000 cells per milliliter or an optical density of 0.180 at 550 nanometers. Sample the culture at regular intervals of 12 hours for Chlorella sorokiniana and eight hours for Haematococcus pluvialis.Place 40 milliliters of the sample into a plastic conical centrifugation tube. Centrifuge the tube at 3000 G for 20 minutes at 15 degrees Celsius. Decant the supernatant and add three milliliters of 90%pure acetone solution to resuspend the cells, then transfer the cells into a glass tube covered with aluminum foil to prevent oxidation and mix using a vortex.Sonicate the suspended sample in an ice bath for two cycles of five minutes each, and let the sample rest at four degrees Celsius for 16 hours. After resting, sonicate the sample again for two cycles of five minutes each under the same conditions, and centrifuge the sample at 3000 G for 20 minutes at 15 degrees Celsius. Separate the pigment extract using a Pasteur pipette and transfer it to another clean tube protected from light.Then place the pigment extract in a quartz cell and read it in a spectrophotometer at 600, 647, and 664 nanometers. Calculate chlorophyll concentrations using the equation. Chlorophyll content of Chlorella sorokiniana was highest in treatment four, under conditions of high carbon dioxide addition, purple light, and high light intensity.Main effects plot showed that light intensity had a lower effect compared to carbon dioxide and purple light effects. In Chlorella sorokiniana, a consistent increase in chlorophylls A and B was seen throughout the experiment, with chlorophyll A surpassing chlorophyll B at 50 hours. A decrease in chlorophyll concentration was observed in Haematococcus pluvialis at 50 hours, with chlorophyll B decreasing significantly while chlorophyll A remained higher.

evaluating-chlorophyll-content-chlorella-sorokiniana-haematococcus

Research

JoVE Journal

Biochemistry

259 Views. To begin, prepare the inoculum to study the growth kinetics of Chlorella sorokiniana and Haematococcus pluvialis Prepare the required volume of sterile culture medium according to standard laboratory procedures. Configure the light source for both the photobioreactor and the flask to the desired color spectrum. Cover both systems to prevent external light interference.After adjusting the photoperiod settings, connect the carbon dioxide duration system to the photobioreactor to provide ...