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.

In the recent climate change regime, industrial demand for renewable materials to replace petroleum-derived polymers continues to rise. Of particular interest is polyhydroxybutyrate (PHB) as a substitute for polypropylene. Accumulating evidence indicates that PHB is highly produced as a carbon storage material in various microorganisms. The effects of growth conditions on PHB production have been widely studied in chemolithotrophs, particularly in Rhodobacter. However, the results on PHB production in Rhodobacter have been somewhat inconsistent due to different strains and experimental conditions, and it is currently unclear how diverse environmental factors are linked with PHB production. Here, we report optimized growth conditions for PHB production and show that the growth conditions are closely related to reactive oxygen species (ROS) regulation. PHB accumulates in cells up to approximately 50% at the highest level under dark-aerobic conditions as opposed to light aerobic/anaerobic conditions. According to the time-course, PHB contents increased at 48 h and then gradually decreased. When observing the effect of temperature and medium composition on PHB production, 30 °C and a carbon/nitrogen ratio of 9:1 or more were found to be most effective. Among PHB biosynthetic genes, PhaA and PhaB are highly correlated with PHB production, whereas PhaC and PhaZ showed little change in overall expression levels. We found that, while the amount of hydrogen peroxide in cells under dark conditions was relatively low compared to the light conditions, peroxidase activities and expression levels of antioxidant-related genes were high. These observations suggest optimal culture conditions for growth and PHB production and the importance of ROS-scavenging signaling with regard to PHB production.

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.

Three-dimensionally patterned Ag–Pt alloy catalyst improves a junction quality of electrolyte/SiO_x/Si photocathodes for H₂ evolution.