Optimizing Docosahexaenoic Acid (DHA) Production by Schizochytrium sp. Grown on Waste Glycerol

The aim of this study was to optimize biomass and docosahexaenoic acid (DHA) production by Schizochytrium sp. grown on waste glycerol as an organic carbon source. Parameters having a significant effect on biomass and DHA yields were screened using the fractional Plackett–Burman design and the response surface methodology (RSM). Schizochytrium sp. growth was most significantly influenced by crude glycerin concentration in the growth medium (150 g/dm3), process temperature (27 °C), oxygen in the bioreactor (49.99% v/v), and the concentration of peptone as a source of nitrogen (9.99 g/dm3). The process parameter values identified as optimal for producing high DHA concentrations in the biomass were as follows: glycerin concentration 149.99 g/dm3, temperature 26 °C, oxygen concentration 30% (v/v), and peptone concentration 2.21 g/dm3. The dry cell weight (DCW) obtained under actual laboratory conditions was 66.69 ± 0.66 g/dm3, i.e., 1.27% lower than the predicted value. The DHA concentration obtained in the actual culture was at 17.25 ± 0.33 g/dm3, which was 3.03% lower than the predicted value. The results obtained suggest that a two-step culture system should be employed, with the first phase focused on high production of Schizochytrium sp. biomass, and the second focused on increasing DHA concentration in the cells.

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Effect of the Concentration of Extracellular Polymeric Substances (EPS) and Aeration Intensity on Waste Glycerol Valorization by Docosahexaenoic Acid (DHA) Produced in Heterotrophic Culture of Schizochytrium sp.

Applied Sciences ◽

10.3390/app11209573 ◽

2021 ◽

Vol 11 (20) ◽

pp. 9573

Author(s):

Natalia Kujawska ◽

Szymon Talbierz ◽

Marcin Dębowski ◽

Joanna Kazimierowicz ◽

Marcin Zieliński

Keyword(s):

Docosahexaenoic Acid ◽

Culture Medium ◽

Extracellular Polymeric Substances ◽

Stationary Growth Phase ◽

Operating Costs ◽

Glycerol Conversion ◽

Waste Glycerol ◽

Dry Cell Weight ◽

Dha Production ◽

Dry Cell

The study aimed to determine the effectiveness of docosahexaenoic acid (DHA) production by Schizochytrium sp. biomass fed with waste glycerol depending on the concentration of extracellular polymeric substances (EPS) in the culture medium and medium aeration effectiveness. The microalgae from the genus Schizochytrium sp. were proved to be capable of producing EPS composed of glucose, galactose, mannose, fucose, and xylose. The highest EPS concentration, reaching 8.73 ± 0.09 g/dm3, was determined at the stationary growth phase. A high EPS concentration caused culture medium viscosity to increase, contributing to diminished oxygen availability for cells, lower culture effectiveness, and reduced waste glycerol conversion to DHA. The Schizochytrium sp. culture variant found optimal in terms of the obtained technological effects and operating costs was performed at the volumetric oxygen mass transfer coefficient of kLa = 600 1/h, which enabled obtaining dry cell weight (DCW) of 147.89 ± 4.77 g/dm3, lipid concentration of 69.44 ± 0.76 g/dm3, and DHA concentration in the biomass reaching 29.44 ± 0.36 g/dm3. The effectiveness of waste glycerol consumption in this variant reached 3.76 ± 0.31 g/dm3·h and 3.16 ± 0.22 g/gDCW.

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Cultivation Method Effect on Schizochytrium sp. Biomass Growth and Docosahexaenoic Acid (DHA) Production with the Use of Waste Glycerol as a Source of Organic Carbon

Energies ◽

10.3390/en14102952 ◽

2021 ◽

Vol 14 (10) ◽

pp. 2952

Author(s):

Natalia Kujawska ◽

Szymon Talbierz ◽

Marcin Dębowski ◽

Joanna Kazimierowicz ◽

Marcin Zieliński

Keyword(s):

Fatty Acids ◽

Docosahexaenoic Acid ◽

Batch Culture ◽

Acid Production ◽

Carbon Sources ◽

Fed Batch ◽

Waste Glycerol ◽

Total Fatty Acids ◽

Fed Batch Culture ◽

Dha Production

Inexpensive carbon sources offering an alternative to glucose are searched for to reduce costs of docosahexaenoic acid production by microalgae. The use of waste glycerol seems substantiated and prospective in this case. The objective of this study was to determine the production yield of heterotrophic microalgae Schizochytrium sp. biomass and the efficiency of docosahexaenoic acid production in various types of cultures with waste glycerol. Cultivation conditions were optimized using the Plackett–Burman method and Response Surface Methodology. The highest technological performance was obtained in the fed-batch culture, where the concentration of Schizochytrium sp. biomass reached 103.44 ± 1.50 g/dm3, the lipid concentration in Schizochytrium sp. biomass was at 48.85 ± 0.81 g/dm3, and the docosahexaenoic acid concentration at 21.98 ± 0.36 g/dm3. The highest docosahexaenoic acid content, accounting for 61.76 ± 3.77% of total fatty acids, was determined in lipid bodies of the Schizochytrium sp. biomass produced in the batch culture, whereas the lowest one, accounting for 44.99 ± 2.12% of total fatty acids, in those of the biomass grown in the fed-batch culture.

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Overproduction of docosahexaenoic acid in Schizochytrium sp. through genetic engineering of oxidative stress defense pathways

Biotechnology for Biofuels ◽

10.1186/s13068-021-01918-w ◽

2021 ◽

Vol 14 (1) ◽

Author(s):

Xiao Han ◽

Zhaohui Li ◽

Ying Wen ◽

Zhi Chen

Keyword(s):

Oxidative Stress ◽

Cell Growth ◽

Docosahexaenoic Acid ◽

Peroxidation Of Lipids ◽

Ros Scavenging ◽

Lipid Yield ◽

Oxidative Stress Defense ◽

Dry Cell Weight ◽

Dha Production ◽

Stress Defense

Abstract Background Oxidation and peroxidation of lipids in microorganisms result in increased levels of intracellular reactive oxygen species (ROS) and reactive aldehydes, and consequent reduction of cell growth and lipid accumulation. Results To reduce oxygen-mediated cell damage and increase lipid and docosahexaenoic acid (DHA) production in Schizochytrium sp., we strengthened the oxidative stress defense pathways. Overexpression of the enzymes thioredoxin reductase (TRXR), aldehyde dehydrogenase (ALDH), glutathione peroxidase (GPO), and glucose-6-phosphate dehydrogenase (ZWF) strongly promoted cell growth, lipid yield, and DHA production. Coexpression of ZWF, ALDH, GPO, and TRXR enhanced ROS-scavenging ability. Highest values of dry cell weight, lipid yield, and DHA production (50.5 g/L, 33.1 g/L, and 13.3 g/L, respectively) were attained in engineered strain OaldH-gpo-trxR by shake flask fed-batch culture; these were increases of 18.5%, 80.9%, and 114.5% relative to WT values. Conclusions Our findings demonstrate that engineering of oxidative stress defense pathways is an effective strategy for promoting cell robustness, lipid yield, and DHA production in Schizochytrium.

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