Optimizing cultivation strategies and scaling up for fucoxanthin production using Pavlova sp.

The microalgal-based production of fucoxanthin has emerged as an imperative research endeavor due to its antioxidant, and anticancer properties. In this study, three brown marine microalgae, namely Skeletonema costatum, Chaetoceros gracilis, and Pavlova sp., were screened for fucoxanthin production. All strains displayed promising results, with Pavlova sp. exhibiting the highest fucoxanthin content (27.91 mg/g) and productivity (1.16 mg/L·day). Moreover, the influence of various cultivation parameters, such as culture media, salinity, sodium nitrate concentration, inoculum size, light intensity, and iron concentration, were investigated and optimized, resulting in a maximum fucoxanthin productivity of 7.89 mg/L·day. The investigation was further expanded to large-scale outdoor cultivation using 50 L tubular photobioreactors, illustrating the potential of Pavlova sp. and the cultivation process for future commercialization. The biomass and fucoxanthin productivity for the large-scale cultivation were 70.7 mg/L·day and 4.78 mg/L·day, respectively. Overall, the findings demonstrated considerable opportunities for fucoxanthin synthesis via microalgae cultivation and processing.

Optimizing heterotrophic production of Chlorella sorokiniana SU-9 proteins potentially used as a sustainable protein substitute in aquafeed.

Alternative protein sources for the reduction/replacement of fish meal in aqua-feeds are in urgent demand. Microalgae are considered sustainable protein sources for aquaculture due to their high-quality proteins with a complete profile of essential amino acids. This study examined the heterotrophic production of proteins from Chlorella sorokiniana SU-9. Culture parameters for maximal biomass and protein production are as follows: glucose - 10 g/L glucose, sodium nitrate - 1.5 g/L, and iron - 46 µM iron in BG-11 medium. Under optimal conditions, biomass content, protein content and protein productivity of SU-9 reached 4.14 ± 0.20 g/L, 403 ± 33 mg/g and 382 ± 36 mg/L/d, respectively. The protein profile of Chlorella sorokiniana SU-9 is comparable to fishmeal and soybean meal. The essential amino acids arginine, lysine and cysteine, along with glutamine and glutamate, were high. The production cost of SU-9 can be significantly reduced under heterotrophic cultivation conditions, making it a potential protein substitute in aquafeed.

A novel two-stage process for the effective treatment of swine wastewater using Chlorella sorokiniana AK-1 based algal-bacterial consortium under semi-continuous operation.

This study aimed at developing an eco-friendly and effective treatment for swine wastewater (SWW) using a designer microalgae-bacteria consortium. A functional algal bacterial consortium was developed with SWW-derived bacteria and Chlorella sorokiniana AK-1. Light intensity (300 µmol/m2/s) and inoculum size (0.15 and 0.2 g/L for microalgae and bacteria) were optimized. Semi-batch operation treating 50 % SWW resulted in a COD, BOD, TN, and TP removal efficiency of 81.1 ± 0.9 %, 97.0 ± 0.7 %, 90.6 ± 1.6 % and 91.3 ± 1.1 %, respectively. A novel two-stage process with an initial bacterial start-up stage followed by microalgal inoculation was applied for attaining stable organic carbon removal, in addition to satisfactory TN and TP removal. Full strength SWW was treated with this strategy with COD, BOD, TN, and TP removal efficiencies of 72.1 %, 94.9 %, 88 %, and 94.6 %, respectively. The biomass consisted of 36 % carbohydrates, indicating a potential feedstock for biochar production. In addition, the effluent met the standards for effluent discharge in Taiwan.

Bioethanol production from microalgae biomass at high-solids loadings.

Industrial adoption of microalgae biofuel technology has always been hindered by its economic viability. To increase the feasibility of bioethanol production from microalgae, fermentation was applied to Chlorella vulgaris FSP-E biomass at high-solids loading conditions. First, Chlorella vulgaris FSP-E was cultivated to produce microalgae biomass with high carbohydrate content. Next, different ethanol-producing microorganisms were screened. Saccharomyces cerevisiae FAY-1 showed no inhibition when fermenting high initial glucose concentrations and was selected for the fermentation experiments at high-solids loadings. Optimization of acid hydrolysis at high biomass loading was also performed. The fermentation of microalgal biomass hydrolysate produced a final ethanol concentration and yield higher than most reported literature using microalgae feedstock. In addition, the kinetics of bioethanol fermentation of microalgae hydrolysate under high-solids loading were evaluated. These results showed the potential of fermenting microalgae biomass at high-solids loading in improving the viability of microalgae bioethanol production.

Bioethanol production from Chlorella vulgaris ESP-31 grown in unsterilized swine wastewater.

The potential of microalgae to remove nutrients from swine wastewater and accumulate carbohydrates was examined. Chlorella sorokiniana AK-1 and Chlorella vulgaris ESP-31 were grown in 10% unsterilized swine wastewater and obtained a maximum carbohydrate content and productivity of 42.5% and 189 mg L-1d-1, respectively. At 25% wastewater and 25% BG-11 concentration, the maximum carbohydrate productivity and total nitrogen removal efficiency of C. vulgaris ESP-31 were improved to 266 mg L-1d-1 and 54.2%, respectively. Further modifications in light intensity, inoculum size, and harvesting period enhanced the biomass growth, carbohydrate concentration, and total nitrogen assimilation to 3.6 gL-1, 1.8 gL-1, and 92.2%, respectively. Ethanol fermentation of the biomass resulted in bioethanol yield and concentration of 84.2% and 4.2 gL-1, respectively. Overall, unsterilized swine wastewater was demonstrated as a cost-effective nutrient source for microalgal cultivation which further increases the economic feasibility and environmental compatibility of bioethanol production with concomitant swine wastewater treatment.

Lutein production with Chlorella sorokiniana MB-1-M12 using novel two-stage cultivation strategies - metabolic analysis and process improvement.

Microalgae-derived carotenoids have increasingly been considered as feasible green alternatives for synthetic antioxidants. In this study, the lutein high-yielding strain (Chlorella sorokiniana MB-1; henceforth MB-1) and its mutant derivative (C. sorokiniana MB-1-M12; henceforth M12) were evaluated for their growth, biomass production, and lutein accumulation in three different cultivation modes - photoautotrophy, mixotrophy, and heterotrophy. M12 could grow effectively under heterotrophic conditions, but the lutein content was lower, indicating the necessity of photo-induction for lutein accumulation. Metabolic analysis of MB-1 and M12 in autotrophic growth in the presence of carbon dioxide indicated that carbon assimilation and channeling of the fixed metabolites towards carotenoid accumulation was elevated in M12 compared to MB-1. Novel two-stage alternative cultivation strategies (Autotrophic/Heterotrophic and Mixotrophic/Heterotrophic cultures) were applied for enhancing lutein production in M12. Maximum lutein quantity (6.17 mg/g) and production (33.64 mg/L) were obtained with the TSHM strategy that is considered the best two-stage operation.

A novel process for the mixotrophic production of lutein with Chlorella sorokiniana MB-1-M12 using aquaculture wastewater.

In this study, microalgal cultivation was applied as a feasible strategy for treating shrimp culture wastewater (SCW) from a shrimp farm in southern Tainan. Chlorella sorokiniana MB-1-M12 was first grown on BG-11 medium with 0.5% salinity, obtaining a biomass concentration and productivity of 4.35 g/L and 1.56 g/L/d, respectively. When 80% of BG-11 nutrients were added to 75% strength SCW, lutein content and productivity increased to 5.19 mg/g and 5.55 mg/L/d, respectively. A novel operation strategy involving periodic exchange of freshwater and SCW was designed for semi-continuous cultivation of MB-1-M12 strain for optimal biomass and lutein production. The average biomass concentration, productivity, lutein content, and productivity were 3.5 g/L, 1.3 g/L/d, 3.89 mg/g and 5.0 mg/L/d, respectively. Although microalgae have been considered as an alternative natural source of lutein, this work is among the earliest reports describing lutein production from microalgae cultivated with wastewater via a circular economy concept.

Enhancing lutein production with mixotrophic cultivation of Chlorella sorokiniana MB-1-M12 using different bioprocess operation strategies.

A lutein-enriched mutant, Chlorella sorokiniana MB-1-M12 was grown mixotrophically for lutein production. The lutein production efficiency of the strain was enhanced via optimizing the operating strategies. The results show that using semi-continuous cultivation with a medium replacement ratio of 75% resulted in a higher lutein productivity and lutein concentration of 6.24 mg/L/d and 50.6 mg/L, respectively, which were markedly higher than those obtained from batch and fed-batch cultivation. Cultivation under simulated outdoor cultivation conditions (i.e., temperature of 35 °C/25 °C for a 12 h/12 h light/dark cycle) could achieve the highest lutein productivity and lutein concentration of 3.34 mg/L/d and 30.8 mg/L, respectively. Lutein production via outdoor cultivation of MB-1-M12 strain with a 60-L tubular photobioreactor was performed using semi-continuous operation. With a medium replacement ratio of 75%, a good lutein productivity (4.46 mg/L/d) and concentration (27.4 mg/L) was obtained, indicating the feasibility of producing lutein under outdoor cultivation of the microalgal strain.