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Guangzhou Scikoon Industry Ltd. Co.
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2024 Technical Program

Biotechnology

Protein and Co-Products

Processing

Health and Nutrition

Session: Fermentation II: Precision Fermentation of Alternative Oils With Unique Health Benefits

Production of Human Milk Fat Analogue by Fermentation of Microalgae

Tuesday, April 30, 2024
11:35 AM – 11:55 AM EDT
Room: 524b
Abstract: We developed a fermentation technology to produce essential human milk triacylglycerols (TAG), addressing a critical challenge in infant nutrition. Extensive literature analysis guided the selection of specific TAG targets. Leveraging a microalgal oil production platform, we implemented a two-step procedure to modify fatty acid and TAG composition. Classical strain improvement techniques increased palmitic acid (16:0) content, followed by the expression of a lysophosphatidic acid acyltransferase (LPAAT) sequence, enabling specific esterification of 16:0 at the internal position (sn-2 palmitate) of TAG. Scaling up the fermentation in a 50 L reactor yielded kilograms of biomass, with subsequent extraction and refining using standard oil processing conditions. Liquid chromatography and mass spectrometry monitored TAG profiles and the regiospecificity of 16:0 at the internal position (sn-2 palmitate). The initial strain had a 25% 16:0 level, increased to 30% through classical strain improvement, while oleic acid decreased from 61% to 57%. Introduction of an exogenous LPAAT gene raised 16:0 esterified at the internal position by a factor of 10, reaching >70% of total fatty acids. This reduced oleic acid in the internal position from 81% to 22%, with primary TAG species identified as 1,3-dioleoyl-2-palmitoyl-glycerol (OPO). Large scale fermentation trial confirmed the strain's capability to yield >150 g of oil per liter of fermentation broth in a 5-day timeframe, establishing scalability for larger-scale industrialization. Our work demonstrates the feasibility of producing a suitable TAG composition for integration into infant nutrition formulations, meeting infant feeding requirements when combined with other fats and oils.