Checkerspot Develops Human Milk Fat Analogue by Microalgae Fermentation

Seagriculture EU 2024

Members of the Checkerspot team, left to right: Walter Rakitsky, Dino Athanasiadis, Leon Parker, Jon Wittenberg, Mona Correa, Xiaoying Zhou, Bryce Doherty, Veronica Benites, Xinhua Zhao. Image: Shogo Kawatsu

Checkerspot, based in Alameda, CA, is a biotechnology company and Certified B Corporation™ that produces unique oils through the fermentation of microalgae. Recently they announced in Frontiers in Nutrition the development of a new source of human milk fat analogue known as OPO (Oleic-Palmitic-Oleic or sn-2 palmitate).

Human milk contains a complex mixture of triglycerides, making it challenging to recreate using common ingredients. Currently, infant formula companies use conventional oils such as palm, coconut, canola, and sunflower oils to create formulations that mimic the fatty acid profile found in the lipid fraction of human milk. However, it is incredibly challenging to precisely replicate the composition of human milk triglycerides using only vegetable oils since they do not reflect the unique positioning of the fatty acids on the glycerol backbone.

Checkerspot’s technology allows for precise esterification of specific fatty acids at the targeted position (in vivo), allowing for the desired distribution of triglyceride structures at the molecular level. This results in the production of a new generation of OPO for application in infant nutrition products.

In instances where breastfeeding is impractical due to physiological challenges, personal preferences, or other factors, offering alternatives that mimic the nutritional properties of human milk is the preferred choice for parents and caregivers.

Infant formulas currently available in the market often incorporate structured triglycerides through human milk fat substitutes produced enzymatically. While these first-generation substitutes have notably enhanced the nutritional properties of infant formula, these substitutes exhibit varying levels of sn-2 palmitate, typically ranging from 39% to 65% of the total palmitic acid content. This falls below the level of palmitic acid in the sn-2 position observed in human milk, which is in the range of 70% of total palmitic acid. The variations in these formulations highlight the ongoing efforts in the industry to achieve higher levels of sn-2 palmitate for enhanced health benefits in formula-fed infants.

Checkerspot’s sugars-to-oils molecular biology capability is focused on the development of structured triglycerides and specialty fatty acids that advance nutrition and health. Its molecular foundry is precisely creating carbon chain lengths and specific positioning of fatty acids, while simultaneously scaling up the production of these oils through fermentation.

Producing oils on demand

The ability to produce oils on demand in a controlled environment has the potential to strengthen and secure previously long supply chains and improve ultimate traceability for high purity ingredients.

“Microalgae function very much like higher plants when it comes to lipid metabolism. They typically insert saturated fatty acids at the sn-1 and sn-3 positions on the glycerol backbone and unsaturated fatty acids at the sn-2 position. Successfully making a human milk triglyceride in our system posed a formidable challenge because it goes completely counter to its normal lipid metabolism,” said Scott Franklin, Chief Scientific Officer (CSO) at Checkerspot.

“We obtained a unique human milk fat analog containing palmitic acid in sn-2 position in the range of 73% of total palmitic acid. This achievement marks a significant stride in advancing the field,” he said, recognizing the years of research needed to design a strain capable of producing the triglyceride structures naturally present in human milk.

A typical infant formula contains approximately 25g of lipids per 100g of powder, with 40-60% of this content consisting of a human milk fat analogue as a proper source of structured triglycerides. The remaining portion of lipids includes vegetable oils utilized as a source of essential fatty acids. Producing structured triglycerides for infant nutrition through fermentation enables precise control, delivering the high quality demanded by this category. Moving forward, a comprehensive food safety evaluation will be conducted to comply with the regulatory regimes in various markets.

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Seagriculture USA 2024
Contact Phil Ganz



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