biostudies-literatureShang HQNational Natural Science Foundation of Chinae70018https://www.ebi.ac.uk/biostudies/studies/S-EPMC11968058biostudies-literatureUnknown25(4)Ferulic acid is a high-value chemical synthesized in plants. The ferulic acid biosynthesis is still affected by the insufficient methylation activity of caffeic acid O-methyltransferase (<i>COMT</i>). In this study, we engineered <i>COMT</i> from <i>Arabidopsis thaliana</i> to match caffeic acid, and the mutant <i>COMT</i> ^{N129V-H313A-F174L} showed 4.19-fold enhanced catalytic efficiency for degrading caffeic acid. Then, we constructed the de novo synthesis pathway of ferulic acid by introducing tyrosine ammonia lyase from <i>Flavobacterium johnsoniae</i> (<i>FjTAL</i>), 4-hydroxyphenylacetate 3-hydroxylase from <i>Escherichia coli</i> (<i>EcHpaBC</i>), and mutant <i>COMT</i> ^{N129V-H313A-F174L}, and further increased tyrosine synthesis. Furthermore, we overexpressed twoEngineering in life sciencesEngineering Caffeic Acid O-Methyltransferase for Efficient De Novo Ferulic Acid Synthesis.PMC1196805832172145Chen QHQiang SHu CYZhang CQMeng YHShang HQZheng RBo Yang QfalseEngineering Caffeic Acid O-Methyltransferase for Efficient De Novo Ferulic Acid Synthesis.Ferulic acid is a high-value chemical synthesized in plants. The ferulic acid biosynthesis is still affected by the insufficient methylation activity of caffeic acid O-methyltransferase (<i>COMT</i>). In this study, we engineered <i>COMT</i> from <i>Arabidopsis thaliana</i> to match caffeic acid, and the mutant <i>COMT</i> ^{N129V-H313A-F174L} showed 4.19-fold enhanced catalytic efficiency for degrading caffeic acid. Then, we constructed the de novo synthesis pathway of ferulic acid by introducing tyrosine ammonia lyase from <i>Flavobacterium johnsoniae</i> (<i>FjTAL</i>), 4-hydroxyphenylacetate 3-hydroxylase from <i>Escherichia coli</i> (<i>EcHpaBC</i>), and mutant <i>COMT</i> ^{N129V-H313A-F174L}, and further increased tyrosine synthesis. Furthermore, we overexpressed two2025-01-01T00:00:00Z2025 Apr2026-06-03T04:45:18.618Z2025-07-09T03:04:56.114ZS-EPMC119680584019072710.1002/elsc.70018