Haematococcus pluvialis accumulates astaxanthin, which is a high-value antioxidant, during the red cyst stage of its lifecycle. The development of a rigid cell wall in the cysts hinders the recovery of astaxanthin. We investigated morphological changes and cell disruption of mature H. pluvialis cyst cells while using high-pressure homogenization for astaxanthin extraction. When treated with French-press-cell (pressure, 10,000–30,000 psi; passage, 1–3), the intact cyst cells were significantly broken or fully ruptured, releasing cytoplasmic components, thereby facilitating the separation of astaxanthin by ethyl acetate. Fluorescence microscopy observations using three different fluorescent dyes revealed that a greater degree of cell breakage caused greater external dispersion of astaxanthin, chlorophyll, lipids, proteins, and carbohydrates. The mechanical treatment resulted in a high cell disruption rate of up to 91% based on microscopic cell typing and Coulter methods. After the ethyl acetate extraction, the astaxanthin concentration significantly increased by 15.2 mg/L in proportion to the increase in cell disruption rate, which indicates that cell disruption is a critical factor for solvent-based astaxanthin recovery. Furthermore, this study recommends a synergistic combination of the fast instrumental particle-volume-distribution analysis and microscope-based morphologic phenotyping for the development of practical H. pluvialis biorefinery processes that co-produce various biological products, including lipids, proteins, carbohydrates, chlorophyll, and astaxanthin.

The green microalga Haematococcus pluvialis accumulates astaxanthin, a potent antioxidant pigment, as a defense mechanism against environmental stresses. In this study, we investigated the technical feasibility of a stress-based method for inducing astaxanthin biosynthesis in H. pluvialis using electric stimulation in a two-chamber bioelectrochemical system. When a cathodic (reduction) current of 3 mA (voltage: 2 V) was applied to H. pluvialis cells for two days, considerable lysis and breakage of algal cells were observed, possibly owing to the formation of excess reactive oxygen species at the cathode. Conversely, in the absence of cell breakage, the application of anodic (oxidation) current effectively stimulated astaxanthin biosynthesis at a voltage range of 2–6 V, whereas the same could not be induced in the untreated control. At an optimal voltage of 4 V (anodic current: 30 mA), the astaxanthin content in the cells electro-treated for 2 h was 36.9% higher than that in untreated cells. Our findings suggest that electric treatment can be used to improve astaxanthin production in H. pluvialis culture if bioelectrochemical parameters, such as electric strength and duration, are regulated properly.

To alleviate carbon dioxide (CO₂) emission, alternative approaches have been considered, such as employing microorganisms capable of CO₂-fixation using their own metabolic pathways.

Global warming is currently accelerating due to an increase in greenhouse gas emissions by industrialization. Microbial electrosynthesis (MES) using electroactive autotrophic microorganisms has recently been reported as a method to reduce carbon dioxide, the main culprit of greenhouse gas. However, there are still few cases of application of MES, and the molecular mechanisms are largely unknown. To investigate the growth characteristics in MES, we carried out growth tests according to reducing power sources in Rhodobacter sphaeroides. The growth rate was significantly lower when electrons were directly supplied to cells, compared to when hydrogen was supplied. Through a transcriptome analysis, we found that the expression of reactive oxygen species (ROS)-related genes was meaningfully higher in MES than in normal photoautotrophic conditions. Similarly, endogenous contents of H₂O₂ were higher and peroxidase activities were lower in MES. The exogenous application of ascorbic acid, a representative biological antioxidant, promotes cell growth by decreasing ROS levels, confirming the inhibitory effects of ROS on MES. Taken together, our observations suggest that reduction of ROS by increasing antioxidant activities is important for enhancing the cell growth and production of CO₂-converting substances such as carotenoids in MES in R. sphaeroides