{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,26]],"date-time":"2026-07-26T05:56:16Z","timestamp":1785045376454,"version":"3.55.0"},"reference-count":31,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2025,5,27]],"date-time":"2025-05-27T00:00:00Z","timestamp":1748304000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000780","name":"European Union\u2019s Horizon 2020 research and innovation program","doi-asserted-by":"publisher","award":["654000"],"award-info":[{"award-number":["654000"]}],"id":[{"id":"10.13039\/501100000780","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Bioengineering"],"abstract":"<jats:p>We report the synthesis of highly enantiopure perdeuterated poly-L-lactic acid and poly-D-lactic acid polymers with well-defined molecular weight by polymerization of perdeuterated lactides. Enantiopure D- and L-lactic acid-d4 monomers were synthesized from sodium pyruvate-d3 using D- and L-lactate dehydrogenase enzymes (D-LDH and L-LDH) as biocatalysts. The reduced form of the co-enzyme nicotinamide adenine dinucleotide-d1 (NADH-d1) was generated in situ from the oxidized form nicotinamide adenine dinucleotide (NAD+) by formate dehydrogenase (FDH)-catalyzed oxidation of sodium formate-d1 to carbon dioxide with concerted reduction of NAD+ to NADH-d1. For the conversion of the perdeuterated lactic acid monomers to the corresponding lactide dimers, we developed a process for generating these compounds in the high purity needed for the final anionic ring-opening polymerization step. This method enabled the generation of a range of perdeuterated polylactic acid polymers that are highly suitable for the characterization of polymer structure and dynamics using neutron scattering, infrared and nuclear magnetic resonance spectroscopy methods that are sensitive to deuterium. Furthermore, these deuterium-labeled polymers are well-suited to the study of the biodegradation of PLA-based plastics.<\/jats:p>","DOI":"10.3390\/bioengineering12060575","type":"journal-article","created":{"date-parts":[[2025,5,28]],"date-time":"2025-05-28T08:12:47Z","timestamp":1748419967000},"page":"575","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["The Enzymatic Synthesis of Perdeuterated D- and L-Lactic Acid-d4 and Polymerization of Their Lactides to Polylactic Acid"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8196-9774","authenticated-orcid":false,"given":"Anna E.","family":"Leung","sequence":"first","affiliation":[{"name":"European Spallation Source ERIC, P.O. Box 176, 221 00 Lund, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andreas","family":"Raba","sequence":"additional","affiliation":[{"name":"J\u00fclich Centre for Neutron Science (JCNS-1), Forschungszentrum J\u00fclich GmbH, Leo Brandt Stra\u00dfe, 52425 J\u00fclich, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Klaus","family":"Beckerle","sequence":"additional","affiliation":[{"name":"Institute for Inorganic Chemistry, RWTH Aachen University, 52074 Aachen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J\u00fcrgen","family":"Allgaier","sequence":"additional","affiliation":[{"name":"J\u00fclich Centre for Neutron Science (JCNS-1), Forschungszentrum J\u00fclich GmbH, Leo Brandt Stra\u00dfe, 52425 J\u00fclich, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5248-1147","authenticated-orcid":false,"given":"Hanna P.","family":"Wacklin-Knecht","sequence":"additional","affiliation":[{"name":"European Spallation Source ERIC, P.O. Box 176, 221 00 Lund, Sweden"},{"name":"Division of Physical Chemistry, Department of Chemistry, Lund University, 221 00 Lund, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,5,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Jim\u00e9nez, A., Peltzer, M., and Ruseckaite, R. (2014). PLA Synthesis. From the Monomer to the Polymer. Poly(lactic Acid) Science and Technology: Processing, Properties, Additives and Applications, The Royal Society of Chemistry.","DOI":"10.1039\/9781782624806"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"904","DOI":"10.1021\/ma00170a034","article-title":"Stereocomplex Formation Between Enantiomeric Poly(Lactides)","volume":"20","author":"Ikada","year":"1987","journal-title":"Macromolecules"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1123","DOI":"10.1016\/S0079-6700(02)00012-6","article-title":"Properties of Lactic Acid Based Polymers and Their Correlation with Composition","volume":"27","author":"Stolt","year":"2002","journal-title":"Prog. Polym. Sci."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1857","DOI":"10.1021\/ma0357523","article-title":"Melt Chain Dimensions of Polylactide","volume":"37","author":"Anderson","year":"2004","journal-title":"Macromolecules"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"3022","DOI":"10.1002\/anie.201708903","article-title":"C\u2212H Functionalisation for Hydrogen Isotope Exchange","volume":"57","author":"Atzrodt","year":"2018","journal-title":"Angew. Chem. Int. Ed."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"5601","DOI":"10.1021\/ac500952z","article-title":"Parahydrogen Induced Polarization of 1-13C-Phospholactate-D2 for Biomedical Imaging with >30,000,000-Fold NMR Signal Enhancement in Water","volume":"86","author":"Shchepin","year":"2014","journal-title":"Anal Chem."},{"key":"ref_7","first-page":"47","article-title":"Vicinal Deuterium Isotope Effects on Proton Chemical Shifts","volume":"2","author":"Saur","year":"1970","journal-title":"J. Magn. Reson."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"495","DOI":"10.1016\/S0021-9258(18)73943-4","article-title":"The Preparation of Fatty Acids Containing Deuterium","volume":"125","author":"Rittenberg","year":"1938","journal-title":"J. Biol. Chem."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1511","DOI":"10.1139\/v67-245","article-title":"A Simplified Preparation of Fully Deuterated, High Molecular Weight Hydrocarbons","volume":"45","author":"Atkinson","year":"1967","journal-title":"Can. J. Chem."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2215","DOI":"10.1002\/adsc.200800407","article-title":"A Convenient and Effective Method for the Regioselective Deuteration of Alcohols","volume":"350","author":"Maegawa","year":"2008","journal-title":"Adv. Synth. Catal."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1454","DOI":"10.1038\/s41467-020-15310-z","article-title":"Bringing Biocatalytic Deuteration into the Toolbox of Asymmetric Isotopic Labelling Techniques","volume":"11","author":"Rowbotham","year":"2020","journal-title":"Nat. Commun."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2959","DOI":"10.1021\/ja00217a044","article-title":"L-Lactate Dehydrogenase: Substrate Specificity and Use as a Catalyst in the Synthesis of Homochiral 2-Hydroxy Acids","volume":"110","author":"Kim","year":"1988","journal-title":"J. Am. Chem. Soc."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"302","DOI":"10.1021\/ja00469a062","article-title":"Immobilization of Synthetically Useful Enzymes by Condensation Polymerization","volume":"100","author":"Pollak","year":"1978","journal-title":"J. Am. Chem. Soc."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1007\/BF02921743","article-title":"D-Lactate Dehydrogenase","volume":"22","author":"Simon","year":"1989","journal-title":"Appl. Biochem. Biotechnol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1283","DOI":"10.1021\/ja00238a067","article-title":"Membrane-Enclosed Enzymic Catalysis (MEEC): A Useful, Practical New Method for the Manipulation of Enzymes in Organic Synthesis","volume":"109","author":"Bednarski","year":"1987","journal-title":"J. Am. Chem. Soc."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"400","DOI":"10.1016\/0045-2068(89)90041-2","article-title":"Lactate Dehydrogenase-Catalyzed Regeneration of NAD from NADH for Use in Enzyme-Catalyzed Synthesis","volume":"17","author":"Whitesides","year":"1989","journal-title":"Bioorg. Chem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1007\/BF02798431","article-title":"Regeneration of Nicotinamide Cofactors for Use in Organic Synthesis","volume":"14","author":"Chenault","year":"1987","journal-title":"Appl. Biochem. Biotechnol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"326","DOI":"10.1039\/c39910000326","article-title":"Synthesis of Optically Pure (R)-2-Hydroxy Acids Using D-Lactate Dehydrogenase","volume":"5","author":"Kim","year":"1991","journal-title":"J. Chem. Soc. Chem. Commun."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"5012","DOI":"10.1021\/ja00353a026","article-title":"Enzyme-Catalyzed Organic Synthesis: Regeneration of Deuterated Nicotinamide Cofactors for Use in Large-Scale Enzymatic Synthesis of Deuterated Substances","volume":"105","author":"Wong","year":"1983","journal-title":"J. Am. Chem. Soc."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1002\/jcp.1030470414","article-title":"Steric Specificity of Hydrogen Transfer in Pyridine Nucleotide Dehydrogenase Reactions","volume":"47","author":"Vennesland","year":"1956","journal-title":"J. Cell Comp. Physiol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"907","DOI":"10.1002\/cssc.201501695","article-title":"Lactide Synthesis and Chirality Control for Polylactic Acid Production","volume":"9","author":"Dusselier","year":"2016","journal-title":"Chem. Sus. Chem."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"486","DOI":"10.1039\/B815104K","article-title":"Stereocontrolled Ring-Opening Polymerisation of Lactide","volume":"39","author":"Stanford","year":"2010","journal-title":"Chem. Soc. Rev."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"78","DOI":"10.1126\/science.aaa7169","article-title":"Shape-Selective Zeolite Catalysis for Bioplastics Production","volume":"349","author":"Dusselier","year":"2015","journal-title":"Science"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1002\/macp.1964.020750119","article-title":"Preparation of Some Polyesters by Organometallic-Catalyzed Ring Opening Polymerization","volume":"75","author":"Tsuruta","year":"1964","journal-title":"Die Makromol. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1016\/S0045-6535(00)00323-4","article-title":"Syntheses and Application of Polylactides","volume":"43","author":"Kricheldorf","year":"2001","journal-title":"Chemosphere"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1002\/masy.19961030110","article-title":"Polylactides\u2014Synthesis, Characterization and Medical Application","volume":"103","author":"Kricheldorf","year":"1996","journal-title":"Macromol. Symp."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1312","DOI":"10.1002\/adsc.200800788","article-title":"Poly(Trimethylene Carbonate) from Biometals-Based Initiators\/Catalysts: Highly Efficient Immortal Ring-Opening Polymerization Processes","volume":"351","author":"Helou","year":"2009","journal-title":"Adv. Synth. Catal."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"49073","DOI":"10.1002\/app.49073","article-title":"Investigation of Ring-Opening Polymerization of 5-[2-{2-(2-Methoxyethoxy)Ethoxy}-Ethoxymethyl]-5-Methyl-1,3-Dioxa-2-One by Organometallic Catalysts","volume":"137","author":"Chanthaset","year":"2020","journal-title":"J. Appl. Polym. Sci."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"4771","DOI":"10.1002\/pola.20942","article-title":"Synthesis and Characterization of Novel Biodegradable Block Copolymer Poly(Ethylene Glycol)-Block-Poly(L-Lactide-Co-2-Methyl-2-Carboxyl-Propylene Carbonate)","volume":"43","author":"Guan","year":"2005","journal-title":"J. Polym. Sci. A Polym. Chem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"4440","DOI":"10.1021\/acs.analchem.4c05827","article-title":"Raman Microspectroscopy to Trace the Incorporation of Deuterium from Labeled (Micro)Plastics into Microbial Cells","volume":"97","author":"Elsner","year":"2025","journal-title":"Anal Chem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"268","DOI":"10.1016\/j.talanta.2016.11.038","article-title":"Determination of D-Lactide Content in Lactide Stereoisomeric Mixture Using Gas Chromatography-Polarimetry","volume":"164","author":"Feng","year":"2017","journal-title":"Talanta"}],"container-title":["Bioengineering"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2306-5354\/12\/6\/575\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T17:41:25Z","timestamp":1760031685000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2306-5354\/12\/6\/575"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,5,27]]},"references-count":31,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2025,6]]}},"alternative-id":["bioengineering12060575"],"URL":"https:\/\/doi.org\/10.3390\/bioengineering12060575","relation":{},"ISSN":["2306-5354"],"issn-type":[{"value":"2306-5354","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,5,27]]}}}