{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,29]],"date-time":"2026-05-29T17:47:08Z","timestamp":1780076828441,"version":"3.54.0"},"reference-count":34,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2023,6,6]],"date-time":"2023-06-06T00:00:00Z","timestamp":1686009600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100002428","name":"Austrian Science Funds FWF","doi-asserted-by":"publisher","award":["P34934"],"award-info":[{"award-number":["P34934"]}],"id":[{"id":"10.13039\/501100002428","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002428","name":"Austrian Science Funds FWF","doi-asserted-by":"publisher","award":["W1224"],"award-info":[{"award-number":["W1224"]}],"id":[{"id":"10.13039\/501100002428","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Coproheme decarboxylases (ChdCs) are terminal enzymes of the coproporphyrin-dependent heme biosynthetic pathway. In this reaction, two propionate groups are cleaved from the redox-active iron-containing substrate, coproheme, to form vinyl groups of the heme b product. The two decarboxylation reactions proceed sequentially, and a redox-active three-propionate porphyrin, called monovinyl, monopropionate deuteroheme (MMD), is transiently formed as an intermediate. While the reaction mechanism for the first part of the redox reaction, which is initiated by hydrogen peroxide, has been elucidated in some detail, the second part of this reaction, starting from MMD, has not been studied. Here, we report the optimization of enzymatic MMD production by ChdC and purification by reversed-phase chromatography. With the obtained MMD, we were able to study the second part of heme b formation by actinobacterial ChdC from Corynebacterium diphtheriae, starting with Compound I formation upon the addition of hydrogen peroxide. The results indicate that the second part of the decarboxylation reaction is analogous to the first part, although somewhat slower, which is explained by differences in the active site architecture and its H-bonding network. The results are discussed in terms of known kinetic and structural data and help to fill some mechanistic gaps in the overall reaction catalyzed by ChdCs.<\/jats:p>","DOI":"10.3390\/biom13060946","type":"journal-article","created":{"date-parts":[[2023,6,6]],"date-time":"2023-06-06T02:57:30Z","timestamp":1686020250000},"page":"946","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Reactivity of Coproheme Decarboxylase with Monovinyl, Monopropionate Deuteroheme"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4010-4950","authenticated-orcid":false,"given":"Gaurav","family":"Patil","sequence":"first","affiliation":[{"name":"Department of Chemistry, Institute of Biochemistry, University of Natural Resources and Life Sciences, Muthgasse 18, A-1190 Vienna, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hanna","family":"Michlits","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Institute of Biochemistry, University of Natural Resources and Life Sciences, Muthgasse 18, A-1190 Vienna, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1199-2469","authenticated-orcid":false,"given":"Paul G.","family":"Furtm\u00fcller","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Institute of Biochemistry, University of Natural Resources and Life Sciences, Muthgasse 18, A-1190 Vienna, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3375-7715","authenticated-orcid":false,"given":"Stefan","family":"Hofbauer","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Institute of Biochemistry, University of Natural Resources and Life Sciences, Muthgasse 18, A-1190 Vienna, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,6,6]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"118861","DOI":"10.1016\/j.bbamcr.2020.118861","article-title":"Heme biosynthesis in prokaryotes","volume":"1868","author":"Layer","year":"2021","journal-title":"Biochim Biophys Acta Mol. Cell Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"985","DOI":"10.1515\/hsz-2022-0205","article-title":"A primer on heme biosynthesis","volume":"403","author":"Dailey","year":"2022","journal-title":"Biol. Chem."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"2210","DOI":"10.1073\/pnas.1416285112","article-title":"Noncanonical coproporphyrin-dependent bacterial heme biosynthesis pathway that does not use protoporphyrin","volume":"112","author":"Dailey","year":"2015","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"e00048-16","DOI":"10.1128\/MMBR.00048-16","article-title":"Prokaryotic Heme Biosynthesis: Multiple Pathways to a Common Essential Product","volume":"81","author":"Dailey","year":"2017","journal-title":"Microbiol. Mol. Biol. Rev."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1016\/j.abb.2018.01.005","article-title":"Coproheme decarboxylases\u2014Phylogenetic prediction versus biochemical experiments","volume":"640","author":"Pfanzagl","year":"2018","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"4022","DOI":"10.1021\/acs.biochem.5b00492","article-title":"Unusual Peroxide-Dependent, Heme-Transforming Reaction Catalyzed by HemQ","volume":"54","author":"Celis","year":"2015","journal-title":"Biochemistry"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"4386","DOI":"10.1111\/febs.13930","article-title":"Hydrogen peroxide-mediated conversion of coproheme to heme b by HemQ-lessons from the first crystal structure and kinetic studies","volume":"283","author":"Hofbauer","year":"2016","journal-title":"FEBS J."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"6766","DOI":"10.1021\/acscatal.9b00963","article-title":"Redox Cofactor Rotates during Its Stepwise Decarboxylation: Molecular Mechanism of Conversion of Coproheme to Heme","volume":"9","author":"Milazzo","year":"2019","journal-title":"ACS Catal."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"5405","DOI":"10.1021\/acscatal.0c00411","article-title":"Actinobacterial coproheme decarboxylases use histidine as distal base to promote Compound I formation","volume":"10","author":"Michlits","year":"2020","journal-title":"ACS Catal."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"3989","DOI":"10.1074\/jbc.RA117.000830","article-title":"Decarboxylation involving a ferryl, propionate, and a tyrosyl group in a radical relay yields heme","volume":"293","author":"Streit","year":"2018","journal-title":"J. Biol. Chem."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1900","DOI":"10.1021\/jacs.6b11324","article-title":"Structure-Based Mechanism for Oxidative Decarboxylation Reactions Mediated by Amino Acids and Heme Propionates in Coproheme Decarboxylase (HemQ)","volume":"139","author":"Celis","year":"2017","journal-title":"J. Am. Chem. Soc."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Liu, W., Pang, Y., Song, Y., Li, X., Tan, H., and Chen, G. (2022). Reorienting Mechanism of Harderoheme in Coproheme Decarboxylase-A Computational Study. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23052564"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"3600","DOI":"10.1016\/j.bpj.2021.06.042","article-title":"Reaction intermediate rotation during the decarboxylation of coproheme to heme b in C. diphtheriae","volume":"120","author":"Sebastiani","year":"2021","journal-title":"Biophys. J."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2044","DOI":"10.1021\/acs.biochem.8b00186","article-title":"Insights into the Active Site of Coproheme Decarboxylase from Listeria monocytogenes","volume":"57","author":"Milazzo","year":"2018","journal-title":"Biochemistry"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1016\/j.jinorgbio.2019.03.009","article-title":"The hydrogen bonding network of coproheme in coproheme decarboxylase from Listeria monocytogenes: Effect on structure and catalysis","volume":"195","author":"Milazzo","year":"2019","journal-title":"J. Inorg. Biochem."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Sebastiani, F., Baroni, C., Patil, G., Dali, A., Becucci, M., Hofbauer, S., and Smulevich, G. (2023). The Role of the Hydrogen Bond Network in Maintaining Heme Pocket Stability and Protein Function Specificity of C. diphtheriae Coproheme Decarboxylase. Biomolecules, 13.","DOI":"10.3390\/biom13020235"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"e00312","DOI":"10.1042\/BSR20150330","article-title":"From chlorite dismutase towards HemQ\u2014The role of the proximal H-bonding network in heme binding","volume":"36","author":"Hofbauer","year":"2016","journal-title":"Biosci. Rep."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"111718","DOI":"10.1016\/j.jinorgbio.2022.111718","article-title":"An active site at work\u2014The role of key residues in C. diphteriae coproheme decarboxylase","volume":"229","author":"Sebastiani","year":"2022","journal-title":"J. Inorg. Biochem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"890","DOI":"10.1002\/jrs.6326","article-title":"Spectroscopic evidence of the effect of hydrogen peroxide excess on the coproheme decarboxylase from actinobacterial Corynebacterium diphtheriae","volume":"53","author":"Sebastiani","year":"2022","journal-title":"J. Raman Spectrosc."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1021\/acs.biochem.6b00958","article-title":"Reactions of Ferrous Coproheme Decarboxylase (HemQ) with O2 and H2O2 Yield Ferric Heme b","volume":"56","author":"Streit","year":"2017","journal-title":"Biochemistry"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"5398","DOI":"10.1021\/acs.biochem.6b00701","article-title":"Chemistry and Molecular Dynamics Simulations of Heme b-HemQ and Coproheme-HemQ","volume":"55","author":"Hofbauer","year":"2016","journal-title":"Biochemistry"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"5259","DOI":"10.1021\/acs.biochem.9b00140","article-title":"Control of Metabolite Flux during the Final Steps of Heme b Biosynthesis in Gram-Positive Bacteria","volume":"58","author":"Celis","year":"2019","journal-title":"Biochemistry"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1016\/j.abb.2015.01.010","article-title":"Structure and heme-binding properties of HemQ (chlorite dismutase-like protein) from Listeria monocytogenes","volume":"574","author":"Hofbauer","year":"2015","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"140536","DOI":"10.1016\/j.bbapap.2020.140536","article-title":"Understanding molecular enzymology of porphyrin-binding alpha + beta barrel proteins\u2014One fold, multiple functions","volume":"1869","author":"Hofbauer","year":"2021","journal-title":"Biochim. Biophys. Acta Proteins Proteom."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"807678","DOI":"10.3389\/fbioe.2021.807678","article-title":"Initial Steps to Engineer Coproheme Decarboxylase to Obtain Stereospecific Monovinyl, Monopropionyl Deuterohemes","volume":"9","author":"Michlits","year":"2022","journal-title":"Front. Bioeng. Biotechnol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"543","DOI":"10.1016\/0006-3002(59)90407-X","article-title":"Cleavage of the haem-protein link by acid methylethylketone","volume":"35","author":"Teale","year":"1959","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1179\/135100000101535717","article-title":"The reactivity of myeloperoxidase compound I formed with hypochlorous acid","volume":"5","author":"Burner","year":"2000","journal-title":"Redox Rep."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"5425","DOI":"10.1021\/acs.biochem.5b00831","article-title":"Eukaryotic Catalase-Peroxidase: The Role of the Trp-Tyr-Met Adduct in Protein Stability, Substrate Accessibility, and Catalysis of Hydrogen Peroxide Dismutation","volume":"54","author":"Gasselhuber","year":"2015","journal-title":"Biochemistry"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"14823","DOI":"10.1074\/jbc.RA118.004773","article-title":"Roles of distal aspartate and arginine of B-class dye-decolorizing peroxidase in heterolytic hydrogen peroxide cleavage","volume":"293","author":"Pfanzagl","year":"2018","journal-title":"J. Biol. Chem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"725","DOI":"10.1002\/jrs.1037","article-title":"Relationship between heme vinyl conformation and the protein matrix in peroxidases","volume":"34","author":"Marzocchi","year":"2003","journal-title":"J. Raman Spectrosc."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"796","DOI":"10.1073\/pnas.94.3.796","article-title":"Hydrogen peroxide-mediated alteration of the heme prosthetic group of metmyoglobin to an iron chlorin product: Evidence for a novel oxidative pathway","volume":"94","author":"Sugiyama","year":"1997","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"8199","DOI":"10.1016\/S0021-9258(19)70630-9","article-title":"The stereochemistry of peroxidase catalysis","volume":"255","author":"Poulos","year":"1980","journal-title":"J. Biol. Chem."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"575","DOI":"10.1016\/S0021-9258(19)86214-2","article-title":"The crystal structure of cytochrome c peroxidase","volume":"255","author":"Poulos","year":"1980","journal-title":"J. Biol. Chem."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"3919","DOI":"10.1021\/cr400415k","article-title":"Heme enzyme structure and function","volume":"114","author":"Poulos","year":"2014","journal-title":"Chem. Rev."}],"container-title":["Biomolecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-273X\/13\/6\/946\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T19:49:01Z","timestamp":1760125741000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-273X\/13\/6\/946"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,6,6]]},"references-count":34,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2023,6]]}},"alternative-id":["biom13060946"],"URL":"https:\/\/doi.org\/10.3390\/biom13060946","relation":{},"ISSN":["2218-273X"],"issn-type":[{"value":"2218-273X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,6,6]]}}}