and a symmetrical C10-dialdehyde has been discovered and is used in industrial production. It combines these chemicals together with an ethynylation and then a Wittig reaction. Two equivalents of the proper ylide combined with the proper dialdehyde in a solvent of methanol, ethanol, or a mixture of the two, yields astaxanthin in up to 88% yields. Metabolic engineering The cost of astaxanthin extraction, high market price, and lack of efficient fermentation production systems, combined with the intricacies of chemical synthesis, discourage its commercial development. The metabolic engineering of bacteria (Escherichia coli) enables efficient astaxanthin production from beta-carotene via either zeaxanthin or canthaxanthin. Structure Stereoisomers In addition to structural isomeric configurations, astaxanthin also contains two chiral centers at the 3- and 3-positions, resulting in three unique stereoisomers (3R,3R and 3R,3'S meso and 3S,3'S). While all three stereoisomers are present in nature, relative distribution varies considerably from one organism to another. Synthetic astaxanthin contains a mixture of all three stereoisomers, in approximately 1:2:1 proportions. Esterification Astaxanthin exists in two predominant forms, non-esterified (yeast, synthetic) or esterified (algal) with various length fatty acid moieties whose composition is influenced by the source organism as well as growth conditions. The astaxanthin fed to salmon to enhance flesh coloration is in the non-esterified form The predominance of evidence supports a de-esterification of fatty acids from the astaxanthin molecule in the intestine prior to or concomitant with absorption resulting in the circulation and tissue deposition of non-esterified astaxanthin. European Food Safety Authority (EFSA) published a scientific opinion on a similar xanthophyll carotenoid, lutein, stating that "following passage through the gastrointestinal tract and/or uptake lutein esters are hydrolyzed to form free lutein again". While it can be assumed that non-esterified astaxanthin would be more bioavailable than esterified astaxanthin due to the extra enzymatic steps in the intestine needed to hydrolyse the fatty acid components, several studies suggest that bioavailability is more dependent on formulation than configuration. Uses Astaxanthin is used as a dietary supplement and feed supplement as food colorant for salmon, crabs, shrimp, chickens and egg production. For seafood and animals The primary use of synthetic astaxanthin today is as an animal feed additive to impart coloration, including farm-raised salmon and chicken egg yolks. Synthetic carotenoid pigments colored yellow, red or orange represent about 15–25% of the cost of production of commercial salmon feed. In the 21st century, most commercial astaxanthin for aquaculture is produced synthetically. Class