Formaldehyde formation in the glycine cleavage system and its use for an aldolase-based biosynthesis of 1,3-prodanediol

Formaldehyde formation in the glycine cleavage system and its use for an aldolase-based biosynthesis of 1,3-prodanediol

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Abstract Glycine cleavage system (GCS) occupies a key position in one-carbon (C1) metabolic pathway Misc robotics mecha and receives great attention for the use of C1 carbons like formate and CO2 via synthetic biology.In this work, we demonstrate that formaldehyde exists as a substantial byproduct of the GCS reaction cycle.Three causes are identified for its formation.First, the principal one is the decomposition of N 5 ,N 10 -methylene-tetrahydrofolate (5,10-CH2-THF) to form formaldehyde and THF.Increasing the rate of glycine cleavage promotes the formation of 5,10-CH2-THF, thereby increasing the formaldehyde release rate.

Next, formaldehyde can be produced in the GCS even in the absence of THF.The reason is that T-protein of the GCS can degrade methylamine-loaded H-protein (Hint) to formaldehyde and ammonia, accompanied with the formation of dihydrolipoyl H-protein (Hred), but the reaction rate is less than 0.16% of that in the presence of THF.Increasing T-protein Toy Playsets concentration can speed up the release rate of formaldehyde by Hint.Finally, a certain amount of formaldehyde can be formed in the GCS due to oxidative degradation of THF.

Based on a formaldehyde-dependent aldolase, we elaborated a glycine-based one carbon metabolic pathway for the biosynthesis of 1,3-propanediol (1,3-PDO) in vitro.This work provides quantitative data and mechanistic understanding of formaldehyde formation in the GCS and a new biosynthetic pathway of 1,3-PDO.