Method for the preparation of 2,4-dihydroxybutyric acid (2,4-DHB) comprising the successive steps of: a) a first step of converting malate, and/or succinyl-CoA and/or glyoxylate into malyl-CoA, b) a second step of converting malyl-CoA previously obtained into malate-4-semialdehyde, c) a third step of converting malate-4-semialdehyde into 2,4-DHB using a DHB dehydrogenase; wherein - step a) is catalyzed by an enzyme having malyl-CoA synthetase, succinyl-CoA: -malate-CoA transferase and/or malyl-CoA lyase activity, respectively; said enzymes being chosen among malate-coenzyme A ligase as defined by EC 6.2.1.9, succinyl-CoA -malate CoA transferase as defined by EC 2.8.3.- or malyl-CoA lyase as defined by EC 4.1.3.24, functional variant or fragment thereof; - step b) is catalyzed by an enzyme having malyl-CoA reductase activity, the enzyme having malyl-CoA reductase activity is chosen among malonyl-CoA reductase as defined by EC 1.2.1.75, succinyl-CoA reductase as defined by EC 1.2.1.76, or reported 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase as defined by EC 1.1.1.38 and EC 1.1.1.88, cinnamoyl-CoA reductase as defined by EC 1.2.1.44, or acetaldehyde dehydrogenase as defined by EC 1.2.1.10, or any mutant improving activity and/or substrate affinity of mutated enzyme for malyl-CoA or decreasing its activity on the natural substrate; - step c) is catalyzed by the methylbutyraldehyde reductase Ypr1 of S. cerevisiae, the succinic semialdehyde reductase of M. sedula, the 4-hydroxybutyrate dehydrogenase 4hbd of P. gingivalis, or the alcohol dehydrogenase Yqhd of E. coli any mutant improving activity and/or substrate affinity of mutated enzyme for malate-4-semi aldehyde and/or reducing its activity or affinity for its natural substrate by at least a factor 2. a) a first step of converting malate, and/or succinyl-CoA and/or glyoxylate into malyl-CoA, b) a second step of converting malyl-CoA previously obtained into malate-4-semialdehyde, c) a third step of converting malate-4-semialdehyde into 2,4-DHB using a DHB dehydrogenase; - step a) is catalyzed by an enzyme having malyl-CoA synthetase, succinyl-CoA: -malate-CoA transferase and/or malyl-CoA lyase activity, respectively; said enzymes being chosen among malate-coenzyme A ligase as defined by EC 6.2.1.9, succinyl-CoA -malate CoA transferase as defined by EC 2.8.3.- or malyl-CoA lyase as defined by EC 4.1.3.24, functional variant or fragment thereof; - step b) is catalyzed by an enzyme having malyl-CoA reductase activity, the enzyme having malyl-CoA reductase activity is chosen among malonyl-CoA reductase as defined by EC 1.2.1.75, succinyl-CoA reductase as defined by EC 1.2.1.76, or reported 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase as defined by EC 1.1.1.38 and EC 1.1.1.88, cinnamoyl-CoA reductase as defined by EC 1.2.1.44, or acetaldehyde dehydrogenase as defined by EC 1.2.1.10, or any mutant improving activity and/or substrate affinity of mutated enzyme for malyl-CoA or decreasing its activity on the natural substrate; - step c) is catalyzed by the methylbutyraldehyde reductase Ypr1 of S. cerevisiae, the succinic semialdehyde reductase of M. sedula, the 4-hydroxybutyrate dehydrogenase 4hbd of P. gingivalis, or the alcohol dehydrogenase Yqhd of E. coli any mutant improving activity and/or substrate affinity of mutated enzyme for malate-4-semi aldehyde and/or reducing its activity or affinity for its natural substrate by at least a factor 2.