Table 1 Theoretical maximum yields of 2,4-dihydroxybutyrate (DHB) on methanol (MeOH) and ethylene glycol (EG) according to our synthetic pathway or alternative routes

Subst.	Pathways	Theoretical yield (mol/mol)	Theoretical yield (g/g)	O₂ demand (mol/mol)
MeOH	RuMP^a + malyl-P^b	0.22	0.83	2.8
	RumP^a + malyl-CoA^c	0.17	0.64	5.0
	Synthetic pathway	0.25	0.94	2.0
EG	Tartronic SA^d+ malyl-P^b	0.33	0.64	3.5
	Tartronic SA^d + malyl-CoA^c	0.33	0.64	3.5
	β-hydroxyaspartate cycle^e + malyl-P^b	0.33	0.64	3.5
	β-hydroxyaspartate cycle^e + malyl-CoA^c	0.33	0.64	3.5
	Synthetic pathway	0.50	0.97	1.0

^aNatural methanol assimilation pathway²⁷ via ribulose-5P (RuMP).
^bNon-natural DHB pathway²¹ derived from L-malate natural precursor via malyl-P intermediate.
^cNon-natural DHB pathway²⁴ derived from glyoxylate/acetyl-CoA natural precursors via malyl-CoA intermediate.
^dNatural EG assimilation pathway¹⁵ via tartronic semialdehyde.
^eEG assimilation pathway via β-hydroxyaspartate cycle which yields oxaloacetate from glyoxylate in 4 reactions²³.

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