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Improved High-Throughput Raman-Activated Cell Sorting Yields a High-DHA Aurantiochytrium Mutant with Enhanced Lipid
Sen Wang1,2,3,4, Xixian Wang1,2,3,4, Caili Sun1,2,3
1CAS Key Laboratory of Biofuels, Shandong Provincial Key Laboratory of Energy Genetics, Shandong Engineering Research Center of Single Cell Oil, Qingdao Engineering Laboratory of Single Cell Oil, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao 266101, Shandong, China.
High-throughput Raman-activated cell sorting enables rapid screening of microbial strains for docosahexaenoic acid (DHA) production. This method identified a superior strain, ABBS26, with over 65% DHA content for potential food and pharmaceutical applications.
Area of Science:
- Biotechnology
- Microbial strain development
- Bioprocess engineering
Background:
- Industrial microbial strain breeding is limited by low-throughput phenotypic screening.
- Efficient production of docosahexaenoic acid (DHA) is crucial for food and pharmaceutical applications.
- Thraustochytrids are key microorganisms for producing polyunsaturated fatty acids (PUFAs).
Purpose of the Study:
- To develop a high-throughput screening method for microbial DHA production.
- To isolate and characterize a high-DHA-producing microbial strain.
- To optimize fermentation processes for enhanced DHA yield.
Main Methods:
- Utilized Raman-activated cell sorting for high-throughput intracellular DHA quantification.
- Screened a mutant library of 50,000 thraustochytrid cells.
- Performed omics analysis to understand the genetic basis of high DHA content.
- Optimized fed-batch fermentation parameters.
Main Results:
- Isolated strain ABBS26 with over 65% intracellular DHA content.
- Identified robust lipid recycling, including saturated fatty acid targeting, as key to high DHA in ABBS26.
- Achieved a final biomass of 147.3 g/L and DHA production of 47.4 g/L in optimized fermentation.
- Strain ABBS26 reached 67.6% DHA content of total lipids.
Conclusions:
- Raman-activated cell sorting is an effective high-throughput strategy for PUFA-producing microbial strain improvement.
- Strain ABBS26 demonstrates significant potential for industrial-scale DHA production.
- The developed approach accelerates the breeding of enhanced microbial strains for nutraceuticals and pharmaceuticals.
