{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,30]],"date-time":"2026-06-30T22:51:58Z","timestamp":1782859918502,"version":"3.54.5"},"reference-count":115,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2024,1,19]],"date-time":"2024-01-19T00:00:00Z","timestamp":1705622400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>4-Hydroxyphenylacetate 3-hydroxylase (4HPA3H) is a long-known class of two-component flavin-dependent monooxygenases from bacteria, including an oxygenase component (EC 1.14.14.9) and a reductase component (EC 1.5.1.36), with the latter being accountable for delivering the cofactor (reduced flavin) essential for o-hydroxylation. 4HPA3H has a broad substrate spectrum involved in key biological processes, including cellular catabolism, detoxification, and the biosynthesis of bioactive molecules. Additionally, it specifically hydroxylates the o-position of the C4 position of the benzene ring in phenolic compounds, generating high-value polyhydroxyphenols. As a non-P450 o-hydroxylase, 4HPA3H offers a viable alternative for the de novo synthesis of valuable natural products. The enzyme holds the potential to replace plant-derived P450s in the o-hydroxylation of plant polyphenols, addressing the current significant challenge in engineering specific microbial strains with P450s. This review summarizes the source distribution, structural properties, and mechanism of 4HPA3Hs and their application in the biosynthesis of natural products in recent years. The potential industrial applications and prospects of 4HPA3H biocatalysts are also presented.<\/jats:p>","DOI":"10.3390\/ijms25021222","type":"journal-article","created":{"date-parts":[[2024,1,19]],"date-time":"2024-01-19T05:46:44Z","timestamp":1705643204000},"page":"1222","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["4-Hydroxyphenylacetate 3-Hydroxylase (4HPA3H): A Vigorous Monooxygenase for Versatile O-Hydroxylation Applications in the Biosynthesis of Phenolic Derivatives"],"prefix":"10.3390","volume":"25","author":[{"given":"Ping","family":"Sun","sequence":"first","affiliation":[{"name":"The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuping","family":"Xu","sequence":"additional","affiliation":[{"name":"The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuan","family":"Tian","sequence":"additional","affiliation":[{"name":"The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pengcheng","family":"Chen","sequence":"additional","affiliation":[{"name":"The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dan","family":"Wu","sequence":"additional","affiliation":[{"name":"The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pu","family":"Zheng","sequence":"additional","affiliation":[{"name":"The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,1,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"860508","DOI":"10.3389\/fonc.2022.860508","article-title":"Therapeutic Implications of Caffeic Acid in Cancer and Neurological Diseases","volume":"12","author":"Alam","year":"2022","journal-title":"Front. Oncol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"774","DOI":"10.1038\/sj.bjc.6600197","article-title":"The Cancer Preventative Agent Resveratrol Is Converted to the Anticancer Agent Piceatannol by the Cytochrome P450 Enzyme CYP1B1","volume":"86","author":"Potter","year":"2002","journal-title":"Br. J. Cancer"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"863","DOI":"10.1016\/j.ijbiomac.2018.11.146","article-title":"The Attenuation of HIV-1 Tat-Induced Neurotoxicity by Salvianic Acid A and Danshen Granule","volume":"124","author":"Bao","year":"2019","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1039\/D0FO02324H","article-title":"Phenolic Compounds: Current Industrial Applications, Limitations and Future Challenges","volume":"12","author":"Albuquerque","year":"2021","journal-title":"Food Funct."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2162","DOI":"10.1128\/jb.175.7.2162-2167.1993","article-title":"Characterization of an Escherichia coli Aromatic Hydroxylase with a Broad Substrate Range","volume":"175","author":"Prieto","year":"1993","journal-title":"J. Bacteriol."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Guo, W., Zhou, W., Zhou, H., and Chen, X. (2019). Characterization of Enzymatic Properties of Two Novel Enzymes, 3,4-Dihydroxyphenylacetate Dioxygenase and 4-Hydroxyphenylacetate 3-Hydroxylase, from Sulfobacillus acidophilus TPY. BMC Microbiol., 19.","DOI":"10.1186\/s12866-019-1415-9"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"5550","DOI":"10.1046\/j.1432-1033.2001.02490.x","article-title":"A Novel Two-Protein Component Flavoprotein Hydroxylase","volume":"268","author":"Chaiyen","year":"2001","journal-title":"Eur. J. Biochem."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2105","DOI":"10.1128\/AEM.70.4.2105-2109.2004","article-title":"Use of Whole Cells of Pseudomonas aeruginosa for Synthesis of the Antioxidant Hydroxytyrosol via Conversion of Tyrosol","volume":"70","author":"Allouche","year":"2004","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1128\/jb.178.1.111-120.1996","article-title":"Molecular Characterization of the 4-Hydroxyphenylacetate Catabolic Pathway of Escherichia coli W: Engineering a Mobile Aromatic Degradative Cluster","volume":"178","author":"Prieto","year":"1996","journal-title":"J. Bacteriol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"483","DOI":"10.1016\/0304-4165(64)90332-0","article-title":"Metabolism of P-Hydroxyphenylacetic Acid in Pseudomonas Ovalis","volume":"93","author":"Adachi","year":"1964","journal-title":"Biochim. Biophys. Acta (BBA)\u2014Gen. Subj."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1128\/jb.120.1.159-167.1974","article-title":"Bacterial Degradation of 4-Hydroxyphenylacetic Acid and Homoprotocatechuic Acid","volume":"120","author":"Sparnins","year":"1974","journal-title":"J. Bacteriol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"302","DOI":"10.1128\/jb.143.1.302-306.1980","article-title":"Catabolism of 3- and 4-Hydroxyphenylacetate by the 3,4-Dihydroxyphenylacetate Pathway in Escherichia coli","volume":"143","author":"Cooper","year":"1980","journal-title":"J. Bacteriol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"25848","DOI":"10.1016\/S0021-9258(18)35687-4","article-title":"P-Hydroxyphenylacetate-3-Hydroxylase. A Two-Protein Component Enzyme","volume":"267","author":"Arunachalam","year":"1992","journal-title":"J. Biol. Chem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"22823","DOI":"10.1016\/S0021-9258(17)31719-2","article-title":"Molecular Characterization of 4-Hydroxyphenylacetate 3-Hydroxylase of Escherichia coli. A Two-Protein Component Enzyme","volume":"269","author":"Prieto","year":"1994","journal-title":"J. Biol. Chem."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"481","DOI":"10.1128\/AEM.66.2.481-486.2000","article-title":"Characterization of 4-Hydroxyphenylacetate 3-Hydroxylase (HpaB) of Escherichia coli as a Reduced Flavin Adenine Dinucleotide-Utilizing Monooxygenase","volume":"66","author":"Xun","year":"2000","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"4167","DOI":"10.1128\/aem.61.12.4167-4171.1995","article-title":"Polymer Production by Klebsiella pneumoniae 4-Hydroxyphenylacetic Acid Hydroxylase Genes Cloned in Escherichia coli","volume":"61","author":"Gibello","year":"1995","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"160","DOI":"10.1007\/s002030050429","article-title":"Molecular Cloning and Analysis of the Genes Encoding the 4-Hydroxyphenylacetate Hydroxylase from Klebsiella Pneumoniae","volume":"167","author":"Gibello","year":"1997","journal-title":"Arch. Microbiol."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Di Lorenzo, C., Colombo, F., Biella, S., Stockley, C., and Restani, P. (2021). Polyphenols and Human Health: The Role of Bioavailability. Nutrients, 13.","DOI":"10.3390\/nu13010273"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"514","DOI":"10.1039\/C8FO01997E","article-title":"The Effects of Polyphenols and Other Bioactives on Human Health","volume":"10","author":"Fraga","year":"2019","journal-title":"Food Funct."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"104302","DOI":"10.1016\/j.tifs.2023.104302","article-title":"Valorisation of Fruit Waste for Harnessing the Bioactive Compounds and Its Therapeutic Application","volume":"144","author":"Ritika","year":"2023","journal-title":"Trends Food Sci. Technol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2611","DOI":"10.1038\/s41598-020-59451-z","article-title":"Structure-Antioxidant Activity Relationship of Methoxy, Phenolic Hydroxyl, and Carboxylic Acid Groups of Phenolic Acids","volume":"10","author":"Chen","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"433","DOI":"10.1080\/10715760500540442","article-title":"Antioxidant Profile of Dihydroxy- and Trihydroxyphenolic Acids-A Structure\u2013Activity Relationship Study","volume":"40","author":"Siquet","year":"2006","journal-title":"Free Radic. Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"200","DOI":"10.1016\/j.crfs.2021.03.011","article-title":"Extraction of Phenolic Compounds: A Review","volume":"4","author":"Alara","year":"2021","journal-title":"Curr. Res. Food Sci."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"uhab068","DOI":"10.1093\/hr\/uhab068","article-title":"Hydroxylation Decoration Patterns of Flavonoids in Horticultural Crops: Chemistry, Bioactivity, and Biosynthesis","volume":"9","author":"Liu","year":"2022","journal-title":"Hortic. Res."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Heine, T., van Berkel, W., Gassner, G., van P\u00e9e, K.-H., and Tischler, D. (2018). Two-Component FAD-Dependent Monooxygenases: Current Knowledge and Biotechnological Opportunities. Biology, 7.","DOI":"10.3390\/biology7030042"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"670","DOI":"10.1016\/j.jbiotec.2006.03.044","article-title":"Flavoprotein Monooxygenases, a Diverse Class of Oxidative Biocatalysts","volume":"124","author":"Kamerbeek","year":"2006","journal-title":"J. Biotechnol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1016\/j.abb.2013.12.005","article-title":"Flavin Dependent Monooxygenases","volume":"544","author":"Huijbers","year":"2014","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Paul, C.E., Eggerichs, D., Westphal, A.H., Tischler, D., and van Berkel, W.J.H. (2021). Flavoprotein Monooxygenases: Versatile Biocatalysts. Biotechnol. Adv., 51.","DOI":"10.1016\/j.biotechadv.2021.107712"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Pimviriyakul, P., Jaruwat, A., Chitnumsub, P., and Chaiyen, P. (2021). Structural Insights into a Flavin-Dependent Dehalogenase HadA Explain Catalysis and Substrate Inhibition via Quadruple Pi-Stacking. J. Biol. Chem., 297.","DOI":"10.1016\/j.jbc.2021.100952"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"4818","DOI":"10.1074\/jbc.M116.774448","article-title":"Kinetic Mechanism of the Dechlorinating Flavin-Dependent Monooxygenase HadA*","volume":"292","author":"Pimviriyakul","year":"2017","journal-title":"J. Biol. Chem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1016\/bs.enz.2020.05.008","article-title":"Chapter Ten\u2014Phenolic Hydroxylases","volume":"Volume 47","author":"Chaiyen","year":"2020","journal-title":"The Enzymes"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"33107","DOI":"10.1074\/jbc.M703440200","article-title":"Crystal Structure of the Oxygenase Component (HpaB) of the 4-Hydroxyphenylacetate 3-Monooxygenase from Thermus Thermophilus HB8","volume":"282","author":"Kim","year":"2007","journal-title":"J. Biol. Chem."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"718","DOI":"10.1002\/prot.21534","article-title":"Crystal Structure of the Flavin Reductase Component (HpaC) of 4-Hydroxyphenylacetate 3-Monooxygenase from Thermus Thermophilus HB8: Structural Basis for the Flavin Affinity","volume":"70","author":"Kim","year":"2008","journal-title":"Proteins Struct. Funct. Bioinform."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"7087","DOI":"10.1038\/s41598-019-43577-w","article-title":"Structural Insights into Catalytic Versatility of the Flavin-Dependent Hydroxylase (HpaB) from Escherichia coli","volume":"9","author":"Shen","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1002\/cbic.201900277","article-title":"Structural and Functional Characterization of 4-Hydroxyphenylacetate 3-Hydroxylase from Escherichia coli","volume":"21","author":"Deng","year":"2020","journal-title":"ChemBioChem"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1145","DOI":"10.1007\/s00253-013-4958-y","article-title":"Catalytic Activity of the Two-Component Flavin-Dependent Monooxygenase from Pseudomonas Aeruginosa toward Cinnamic Acid Derivatives","volume":"98","author":"Furuya","year":"2014","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"480","DOI":"10.1007\/s00284-007-9016-5","article-title":"Cloning and Characterization of a 4-Hydroxyphenylacetate 3-Hydroxylase From the Thermophile Geobacillus sp. PA-9","volume":"55","author":"Hawumba","year":"2007","journal-title":"Curr. Microbiol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1758","DOI":"10.1099\/mic.0.027839-0","article-title":"From a Consortium Sequence to a Unified Sequence: The Bacillus Subtilis 168 Reference Genome a Decade Later","volume":"155","author":"Barbe","year":"2009","journal-title":"Microbiology"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"589","DOI":"10.1099\/mic.0.031880-0","article-title":"Characterization of the Anthranilate Degradation Pathway in Geobacillus Thermodenitrificans NG80-2","volume":"156","author":"Liu","year":"2010","journal-title":"Microbiology"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"5103","DOI":"10.1021\/bi052313i","article-title":"Crystal Structures of the Short-Chain Flavin Reductase HpaC from Sulfolobus tokodaii Strain 7 in Its Three States: NAD(P)+-Free, NAD+-Bound, and NADP+-Bound","volume":"45","author":"Okai","year":"2006","journal-title":"Biochemistry"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"26435","DOI":"10.1074\/jbc.M501988200","article-title":"A Two-Component Hydroxylase Involved in the Assimilation of 3-Hydroxyphenyl Acetate in Pseudomonas Putida","volume":"280","author":"Sandoval","year":"2005","journal-title":"J. Biol. Chem."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1016\/j.bbaexp.2004.08.003","article-title":"Cloning and Expression of P-Hydroxyphenylacetate 3-Hydroxylase from Acinetobacter baumannii: Evidence of the Divergence of Enzymes in the Class of Two-Protein Component Aromatic Hydroxylases","volume":"1680","author":"Thotsaporn","year":"2004","journal-title":"Biochim. Biophys. Acta (BBA)\u2014Gene Struct. Expr."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1080\/09168451.2015.1072463","article-title":"Biocatalytic Synthesis of 3,4,5,3\u2032,5\u2032-Pentahydroxy-Trans-Stilbene from Piceatannol by Two-Component Flavin-Dependent Monooxygenase HpaBC","volume":"80","author":"Furuya","year":"2016","journal-title":"Biosci. Biotechnol. Biochem."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1177","DOI":"10.1073\/pnas.0608381104","article-title":"Structure of the Monooxygenase Component of a Two-Component Flavoprotein Monooxygenase","volume":"104","author":"Alfieri","year":"2007","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"720","DOI":"10.1107\/S1744309112016909","article-title":"Crystallization and Preliminary X-Ray Analysis of the Reductase Component of p-Hydroxyphenylacetate 3-Hydroxylase from Acinetobacter Baumannii","volume":"68","author":"Oonanant","year":"2012","journal-title":"Acta Crystallogr. Sect. F Struct. Biol. Cryst. Commun."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"10603","DOI":"10.1073\/pnas.0903809106","article-title":"Multiple Pathways Guide Oxygen Diffusion into Flavoenzyme Active Sites","volume":"106","author":"Baron","year":"2009","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"28170","DOI":"10.1074\/jbc.M111.241836","article-title":"Stabilization of C4a-Hydroperoxyflavin in a Two-Component Flavin-Dependent Monooxygenase Is Achieved through Interactions at Flavin N5 and C4a Atoms","volume":"286","author":"Thotsaporn","year":"2011","journal-title":"J. Biol. Chem."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"4492","DOI":"10.1021\/acscatal.5b00439","article-title":"P-Hydroxyphenylacetate 3-Hydroxylase as a Biocatalyst for the Synthesis of Trihydroxyphenolic Acids","volume":"5","author":"Dhammaraj","year":"2015","journal-title":"ACS Catal."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"17044","DOI":"10.1074\/jbc.M512385200","article-title":"Kinetic Mechanisms of the Oxygenase from a Two-Component Enzyme, p-Hydroxyphenylacetate 3-Hydroxylase from Acinetobacter Baumannii","volume":"281","author":"Sucharitakul","year":"2006","journal-title":"J. Biol. Chem."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"960","DOI":"10.1038\/s41467-019-08781-2","article-title":"Promiscuous Enzymatic Activity-Aided Multiple-Pathway Network Design for Metabolic Flux Rearrangement in Hydroxytyrosol Biosynthesis","volume":"10","author":"Chen","year":"2019","journal-title":"Nat. Commun."},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Wang, L., Ma, X., Ruan, H., Chen, Y., Gao, L., Lei, T., Li, Y., Gui, L., Guo, L., and Xia, T. (2021). Optimization of the Biosynthesis of B-Ring Ortho-Hydroxy Lated Flavonoids Using the 4-Hydroxyphenylacetate 3-Hydroxylase Complex (HpaBC) of Escherichia coli. Molecules, 26.","DOI":"10.3390\/molecules26102919"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1016\/j.abb.2018.06.010","article-title":"Crystal Structure of the Flavin Reductase of Acinetobacter Baumannii P-Hydroxyphenylacetate 3-Hydroxylase (HPAH) and Identification of Amino Acid Residues Underlying Its Regulation by Aromatic Ligands","volume":"653","author":"Yuenyao","year":"2018","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"10434","DOI":"10.1021\/bi050615e","article-title":"The Reductase of P-Hydroxyphenylacetate 3-Hydroxylase from Acinetobacter Baumannii Requires p-Hydroxyphenylacetate for Effective Catalysis","volume":"44","author":"Sucharitakul","year":"2005","journal-title":"Biochemistry"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"8611","DOI":"10.1021\/bi7006614","article-title":"Kinetics of a Two-Component p-Hydroxyphenylacetate Hydroxylase Explain How Reduced Flavin Is Transferred from the Reductase to the Oxygenase","volume":"46","author":"Sucharitakul","year":"2007","journal-title":"Biochemistry"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"26213","DOI":"10.1074\/jbc.M112.354472","article-title":"The C-Terminal Domain of 4-Hydroxyphenylacetate 3-Hydroxylase from Acinetobacter Baumannii Is an Autoinhibitory Domain","volume":"287","author":"Phongsak","year":"2012","journal-title":"J. Biol. Chem."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"590","DOI":"10.1016\/j.bbrc.2005.09.081","article-title":"Dynamics Involved in Catalysis by Single-Component and Two-Component Flavin-Dependent Aromatic Hydroxylases","volume":"338","author":"Ballou","year":"2005","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1002\/pro.3525","article-title":"Monooxygenation of Aromatic Compounds by Flavin-Dependent Monooxygenases","volume":"28","author":"Chenprakhon","year":"2019","journal-title":"Protein Sci."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"1481","DOI":"10.1002\/cbic.201900737","article-title":"Creating Flavin Reductase Variants with Thermostable and Solvent-Tolerant Properties by Rational-Design Engineering","volume":"21","author":"Maenpuen","year":"2020","journal-title":"ChemBioChem"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"3865","DOI":"10.1021\/acssynbio.2c00430","article-title":"Engineering a Prokaryotic Non-P450 Hydroxylase for 3\u2032-Hydroxylation of Flavonoids","volume":"11","author":"Wang","year":"2022","journal-title":"ACS Synth. Biol."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"1895","DOI":"10.1002\/bit.25237","article-title":"Biotechnological Production of Plant-Specific Hydroxylated Phenylpropanoids","volume":"111","author":"Lin","year":"2014","journal-title":"Biotechnol. Bioeng."},{"key":"ref_61","doi-asserted-by":"crossref","unstructured":"Herrmann, S., Dippe, M., Pecher, P., Funke, E., Pietzsch, M., and Wessjohann, L.A. (2022). Engineered Bacterial Flavin-Dependent Monooxygenases for the Regiospecific Hydroxylation of Polycyclic Phenols. ChemBioChem, 23.","DOI":"10.1002\/cbic.202100480"},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1016\/j.ymben.2022.08.007","article-title":"Targeting Cofactors Regeneration in Methylation and Hydroxylation for High Level Production of Ferulic Acid","volume":"73","author":"Zhou","year":"2022","journal-title":"Metab. Eng."},{"key":"ref_63","doi-asserted-by":"crossref","unstructured":"Zhang, Q., Jin, Y., Yang, K., Hu, S., Lv, C., Huang, J., Mei, J., Zhao, W., and Mei, L. (2023). Modification of the 4-Hydroxyphenylacetate-3-Hydroxylase Substrate Pocket to Increase Activity towards Resveratrol. Molecules, 28.","DOI":"10.3390\/molecules28145602"},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"19457","DOI":"10.1021\/acs.jafc.3c06479","article-title":"Biosynthesis of 3-Hydroxyphloretin Using Rational Design of 4-Hydroxyphenylacetate 3-Monooxygenase","volume":"71","author":"Xu","year":"2023","journal-title":"J. Agric. Food Chem."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"219","DOI":"10.1007\/s10811-013-0113-5","article-title":"Functional Expression of an Arabidopsis P450 Enzyme, p-Coumarate-3-Hydroxylase, in the Cyanobacterium Synechocystis PCC 6803 for the Biosynthesis of Caffeic Acid","volume":"26","author":"Xue","year":"2014","journal-title":"J. Appl. Phycol."},{"key":"ref_66","first-page":"3466","article-title":"High-density fermentation of Escherichia coli to express 4-hydroxyphenylacetate 3-hydroxylase and efficient biosynthesis of caffeic acid","volume":"38","author":"Zhang","year":"2022","journal-title":"Sheng Wu Gong Cheng Xue Bao"},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"6087","DOI":"10.1128\/AEM.01103-12","article-title":"Biotechnological Production of Caffeic Acid by Bacterial Cytochrome P450 CYP199A2","volume":"78","author":"Furuya","year":"2012","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"1187","DOI":"10.1021\/jf001258b","article-title":"Synthesis of the Antioxidant Hydroxytyrosol Using Tyrosinase as Biocatalyst","volume":"49","author":"Cantos","year":"2001","journal-title":"J. Agric. Food Chem."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"10045","DOI":"10.1073\/pnas.96.18.10045","article-title":"New Routes for Lignin Biosynthesis Defined by Biochemical Characterization of Recombinant Ferulate 5-Hydroxylase, a Multifunctional Cytochrome P450-Dependent Monooxygenase","volume":"96","author":"Humphreys","year":"1999","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"1687","DOI":"10.1021\/cb300222b","article-title":"Regioselective Hydroxylation of Trans-Resveratrol via Inhibition of Tyrosinase from Streptomyces Avermitilis MA4680","volume":"7","author":"Lee","year":"2012","journal-title":"ACS Chem. Biol."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"2853","DOI":"10.1016\/j.tetlet.2014.03.076","article-title":"Regioselective Synthesis of Piceatannol from Resveratrol: Catalysis by Two-Component Flavin-Dependent Monooxygenase HpaBC in Whole Cells","volume":"55","author":"Furuya","year":"2014","journal-title":"Tetrahedron Lett."},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"3288","DOI":"10.1021\/acssynbio.0c00346","article-title":"Efficient Biosynthesis of (2 S)-Eriodictyol from (2 S)-Naringenin in Saccharomyces cerevisiae through a Combination of Promoter Adjustment and Directed Evolution","volume":"9","author":"Gao","year":"2020","journal-title":"ACS Synth. Biol."},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1186\/s12934-022-01815-3","article-title":"Biosynthesis of Eriodictyol from Tyrosine by Corynebacterium Glutamicum","volume":"21","author":"Wu","year":"2022","journal-title":"Microb. Cell Factories"},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1186\/s12934-016-0533-4","article-title":"Hydroxylation of Diverse Flavonoids by CYP450 BM3 Variants: Biosynthesis of Eriodictyol from Naringenin in Whole Cells and Its Biological Activities","volume":"15","author":"Chu","year":"2016","journal-title":"Microb. Cell Factories"},{"key":"ref_75","doi-asserted-by":"crossref","unstructured":"Chu, L.L., Pandey, R.P., Lim, H.N., Jung, H.J., Thuan, N.H., Kim, T.-S., and Sohng, J.K. (2017). Synthesis of Umbelliferone Derivatives in Escherichia coli and Their Biological Activities. J. Biol. Eng., 11.","DOI":"10.1186\/s13036-017-0056-5"},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"6683","DOI":"10.1021\/acs.jafc.0c03156","article-title":"Biocatalytic Production of a Potent Inhibitor of Adipocyte Differentiation from Phloretin Using Engineered CYP102A1","volume":"68","author":"Nguyen","year":"2020","journal-title":"J. Agric. Food Chem."},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"550","DOI":"10.1248\/bpb.b19-00756","article-title":"Equol Inhibits Mushroom Tyrosinase in Vitro through Tight Binding","volume":"43","author":"Niwa","year":"2020","journal-title":"Biol. Pharm. Bull."},{"key":"ref_78","doi-asserted-by":"crossref","unstructured":"Nozawa, D., Matsuyama, A., and Furuya, T. (2022). Biocatalytic Synthesis and Evaluation of Antioxidant and Antibacterial Activities of Hydroxyequols. Bioorganic Med. Chem. Lett., 73.","DOI":"10.1016\/j.bmcl.2022.128908"},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1016\/j.pep.2011.04.013","article-title":"Gene Engineering, Purification, Crystallization and Preliminary X-Ray Diffraction of Cytochrome P450 p-Coumarate-3-Hydroxylase (C3H), the Arabidopsis Membrane Protein","volume":"79","author":"Kim","year":"2011","journal-title":"Protein Expr. Purif."},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"756","DOI":"10.1021\/acssynbio.9b00431","article-title":"De Novo Biosynthesis of Caffeic Acid from Glucose by Engineered Saccharomyces cerevisiae","volume":"9","author":"Li","year":"2020","journal-title":"ACS Synth. Biol."},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1186\/1475-2859-11-42","article-title":"Biosynthesis of Caffeic Acid in Escherichia coli Using Its Endogenous Hydroxylase Complex","volume":"11","author":"Lin","year":"2012","journal-title":"Microb. Cell Factories"},{"key":"ref_82","doi-asserted-by":"crossref","first-page":"3188","DOI":"10.1002\/bit.24988","article-title":"Caffeic Acid Production Enhancement by Engineering a Phenylalanine Over-producing Escherichia coli Strain","volume":"110","author":"Huang","year":"2013","journal-title":"Biotechnol. Bioeng."},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1002\/btpr.2185","article-title":"Optimization of Naringenin and P-Coumaric Acid Hydroxylation Using the Native E. Coli Hydroxylase Complex, HpaBC","volume":"32","author":"Jones","year":"2016","journal-title":"Biotechnol. Prog."},{"key":"ref_84","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1016\/j.eng.2018.11.029","article-title":"Engineering the Biosynthesis of Caffeic Acid in Saccharomyces cerevisiae with Heterologous Enzyme Combinations","volume":"5","author":"Liu","year":"2019","journal-title":"Engineering"},{"key":"ref_85","doi-asserted-by":"crossref","first-page":"377","DOI":"10.1016\/j.foodchem.2012.12.063","article-title":"A New Process for Obtaining Hydroxytyrosol Using Transformed Escherichia coli Whole Cells with Phenol Hydroxylase Gene from Geobacillus Thermoglucosidasius","volume":"139","year":"2013","journal-title":"Food Chem."},{"key":"ref_86","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1016\/j.ymben.2013.06.001","article-title":"Metabolic Engineering of Escherichia coli for Production of Salvianic Acid A via an Artificial Biosynthetic Pathway","volume":"19","author":"Yao","year":"2013","journal-title":"Metab. Eng."},{"key":"ref_87","doi-asserted-by":"crossref","first-page":"7691","DOI":"10.1021\/acs.jafc.0c03065","article-title":"Engineering Escherichia coli for High-Yielding Hydroxytyrosol Synthesis from Biobased l -Tyrosine","volume":"68","author":"Zeng","year":"2020","journal-title":"J. Agric. Food Chem."},{"key":"ref_88","doi-asserted-by":"crossref","first-page":"1499","DOI":"10.1111\/1751-7915.13957","article-title":"Metabolic Engineering of Saccharomyces cerevisiae for Hydroxytyrosol Overproduction Directly from Glucose","volume":"15","author":"Bisquert","year":"2021","journal-title":"Microb. Biotechnol."},{"key":"ref_89","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1016\/j.phytochem.2017.08.016","article-title":"NMR-Based Identification of the Major Bioactive Molecules from an Italian Cultivar of Lycium barbarum","volume":"144","author":"Lopatriello","year":"2017","journal-title":"Phytochemistry"},{"key":"ref_90","first-page":"1038","article-title":"Chemical Constituents of Opuntia Milpa Alta Haw","volume":"23","author":"Luo","year":"2011","journal-title":"Nat. Prod. Res. Dev."},{"key":"ref_91","doi-asserted-by":"crossref","first-page":"695","DOI":"10.1016\/S0031-9422(97)00348-8","article-title":"5-Hydroxyguaiacyl Nuclei as Aromatic Constituents of Native Lignin","volume":"46","author":"Suzuki","year":"1997","journal-title":"Phytochemistry"},{"key":"ref_92","doi-asserted-by":"crossref","unstructured":"Ami\u0107, A., Dimitri\u0107 Markovi\u0107, J.M., Markovi\u0107, Z., Milenkovi\u0107, D., Milanovi\u0107, \u017d., Antonijevi\u0107, M., Masti\u013e\u00e1k Cagardov\u00e1, D., and Rodr\u00edguez-Guerra Pedregal, J. (2021). Theoretical Study of Radical Inactivation, LOX Inhibition, and Iron Chelation: The Role of Ferulic Acid in Skin Protection against UVA Induced Oxidative Stress. Antioxidants, 10.","DOI":"10.3390\/antiox10081303"},{"key":"ref_93","doi-asserted-by":"crossref","first-page":"4797","DOI":"10.1021\/jf800317v","article-title":"Antioxidant Potential of Hydroxycinnamic Acid Glycoside Esters","volume":"56","author":"Kylli","year":"2008","journal-title":"J. Agric. Food Chem."},{"key":"ref_94","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1016\/0014-5793(84)80278-1","article-title":"Ferulic Acid Shydroxylase: A New Cytochrome P-450-Dependent Enzyme from Higher Plant Microsomes Involved in Lignin Synthesis","volume":"169","author":"Grand","year":"1984","journal-title":"Febs Lett."},{"key":"ref_95","doi-asserted-by":"crossref","first-page":"6869","DOI":"10.1073\/pnas.93.14.6869","article-title":"Ferulate-5-Hydroxylase from Arabidopsis Thaliana Defines a New Family of Cytochrome P450-Dependent Monooxygenases","volume":"93","author":"Meyer","year":"1996","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_96","doi-asserted-by":"crossref","first-page":"8955","DOI":"10.1073\/pnas.96.16.8955","article-title":"Coniferyl Aldehyde 5-Hydroxylation and Methylation Direct Syringyl Lignin Biosynthesis in Angiosperms","volume":"96","author":"Osakabe","year":"1999","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_97","doi-asserted-by":"crossref","first-page":"2889","DOI":"10.1021\/acschembio.6b00402","article-title":"A Single-Site Mutation at Ser146 Expands the Reactivity of the Oxygenase Component of p-Hydroxyphenylacetate 3-Hydroxylase","volume":"11","author":"Dhammaraj","year":"2016","journal-title":"ACS Chem. Biol."},{"key":"ref_98","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.abb.2017.03.004","article-title":"Hydroxylation of 4-Hydroxyphenylethylamine Derivatives by R263 Variants of the Oxygenase Component of p-Hydroxyphenylacetate-3-Hydroxylase","volume":"620","author":"Chenprakhon","year":"2017","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_99","doi-asserted-by":"crossref","unstructured":"Chen, S., Wang, X., Cheng, Y., Gao, H., and Chen, X. (2023). A Review of Classification, Biosynthesis, Biological Activities and Potential Applications of Flavonoids. Molecules, 28.","DOI":"10.3390\/molecules28134982"},{"key":"ref_100","doi-asserted-by":"crossref","first-page":"773","DOI":"10.1016\/j.bcp.2004.05.008","article-title":"Involvement of Cytochrome P450 1A2 in the Biotransformation of Trans-Resveratrol in Human Liver Microsomes","volume":"68","author":"Piver","year":"2004","journal-title":"Biochem. Pharmacol."},{"key":"ref_101","doi-asserted-by":"crossref","first-page":"932","DOI":"10.1124\/dmd.108.026484","article-title":"Generation of the Human Metabolite Piceatannol from the Anticancer-Preventive Agent Resveratrol by Bacterial Cytochrome P450 BM3","volume":"37","author":"Kim","year":"2009","journal-title":"Drug Metab. Dispos."},{"key":"ref_102","doi-asserted-by":"crossref","first-page":"248","DOI":"10.1007\/s12010-017-2645-7","article-title":"Enzymatic Activation of the Emerging Drug Resveratrol","volume":"185","author":"Koyani","year":"2018","journal-title":"Appl. Biochem. Biotechnol."},{"key":"ref_103","doi-asserted-by":"crossref","unstructured":"Liu, W., Feng, Y., Yu, S., Fan, Z., Li, X., Li, J., and Yin, H. (2021). The Flavonoid Biosynthesis Network in Plants. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms222312824"},{"key":"ref_104","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1016\/j.abb.2019.04.012","article-title":"Functional Characterization of Flavonoid 3\u2032-Hydroxylase, CsF3\u2032H, from Crocus Sativus L: Insights into Substrate Specificity and Role in Abiotic Stress","volume":"667","author":"Baba","year":"2019","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_105","doi-asserted-by":"crossref","unstructured":"Park, S., Choi, M.J., Lee, J.Y., Kim, J.K., Ha, S.-H., and Lim, S.-H. (2016). Molecular and Biochemical Analysis of Two Rice Flavonoid 3\u2032-Hydroxylase to Evaluate Their Roles in Flavonoid Biosynthesis in Rice Grain. Int. J. Mol. Sci., 17.","DOI":"10.3390\/ijms17091549"},{"key":"ref_106","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1124\/dmd.105.004929","article-title":"Structural elucidation of hydroxylated metabolites of the isoflavan equol by gas chromatography-mass spectrometry and high-performance liquid chromatography-mass spectrometry","volume":"34","author":"Glatt","year":"2006","journal-title":"Drug Metab. Dispos."},{"key":"ref_107","doi-asserted-by":"crossref","unstructured":"Song, H., Lee, P.-G., Kim, J., Kim, J., Lee, S.-H., Kim, H., Lee, U.-J., Kim, J.Y., Kim, E.-J., and Kim, B.-G. (2022). Regioselective One-Pot Synthesis of Hydroxy-(S)-Equols Using Isoflavonoid Reductases and Monooxygenases and Evaluation of the Hydroxyequol Derivatives as Selective Estrogen Receptor Modulators and Antioxidants. Front. Bioeng. Biotechnol., 10.","DOI":"10.3389\/fbioe.2022.830712"},{"key":"ref_108","doi-asserted-by":"crossref","first-page":"21826","DOI":"10.1039\/C9RA03913A","article-title":"Monooxygenase-Catalyzed Regioselective Hydroxylation for the Synthesis of Hydroxyequols","volume":"9","author":"Hashimoto","year":"2019","journal-title":"RSC Adv."},{"key":"ref_109","doi-asserted-by":"crossref","first-page":"1515","DOI":"10.1038\/s41467-020-14918-5","article-title":"Developing a Highly Efficient Hydroxytyrosol Whole-Cell Catalyst by de-Bottlenecking Rate-Limiting Steps","volume":"11","author":"Yao","year":"2020","journal-title":"Nat. Commun."},{"key":"ref_110","doi-asserted-by":"crossref","first-page":"423","DOI":"10.1049\/iet-nbt.2017.0189","article-title":"Encapsulation of the Reductase Component of p-Hydroxyphenylacetate Hydroxylase in Poly(Lactide-Co-Glycolide) Nanoparticles by Three Different Emulsification Techniques","volume":"12","author":"Sawasdee","year":"2018","journal-title":"IET Nanobiotechnol."},{"key":"ref_111","doi-asserted-by":"crossref","first-page":"3163","DOI":"10.1016\/j.ijbiomac.2020.08.182","article-title":"Co-Immobilization of Two-Component Hydroxylase Monooxygenase by Functionalized Magnetic Nanoparticles for Preserving High Catalytic Activity and Enhancing Enzyme Stabilty","volume":"164","author":"Liao","year":"2020","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_112","doi-asserted-by":"crossref","first-page":"563","DOI":"10.1038\/s41467-020-14371-4","article-title":"Compartmentalized Microbes and Co-Cultures in Hydrogels for on-Demand Bioproduction and Preservation","volume":"11","author":"Johnston","year":"2020","journal-title":"Nat. Commun."},{"key":"ref_113","doi-asserted-by":"crossref","first-page":"7490","DOI":"10.1021\/acs.jafc.2c02137","article-title":"De Novo Production of Hydroxytyrosol by Metabolic Engineering of Saccharomyces Cerevisiae","volume":"70","author":"Liu","year":"2022","journal-title":"J. Agric. Food Chem."},{"key":"ref_114","doi-asserted-by":"crossref","first-page":"125646","DOI":"10.1016\/j.foodchem.2019.125646","article-title":"Overproduction of Hydroxytyrosol in Saccharomyces cerevisiae by Heterologous Overexpression of the Escherichia coli 4-Hydroxyphenylacetate 3-Monooxygenase","volume":"308","author":"Bisquert","year":"2020","journal-title":"Food Chem."},{"key":"ref_115","doi-asserted-by":"crossref","unstructured":"Gong, P., Tang, J., Wang, J., Wang, C., and Chen, W. (2023). A Novel Microbial Consortia Catalysis Strategy for the Production of Hydroxytyrosol from Tyrosine. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24086944"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/2\/1222\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T13:45:49Z","timestamp":1760103949000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/2\/1222"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,1,19]]},"references-count":115,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2024,1]]}},"alternative-id":["ijms25021222"],"URL":"https:\/\/doi.org\/10.3390\/ijms25021222","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,1,19]]}}}