{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T18:32:33Z","timestamp":1790274753646,"version":"4.1.0"},"reference-count":90,"publisher":"Wiley","issue":"7","license":[{"start":{"date-parts":[[2021,10,8]],"date-time":"2021-10-08T00:00:00Z","timestamp":1633651200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"content-domain":{"domain":["chemistry-europe.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["ChemBioChem"],"published-print":{"date-parts":[[2022,4,5]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Enantiopure \u03b1\u2010hydroxy ketones are important building blocks of active pharmaceutical ingredients (APIs), which can be produced by thiamine\u2010diphosphate\u2010dependent lyases, such as benzaldehyde lyase. Here we report the discovery of a novel thermostable benzaldehyde lyase from <jats:italic>Rhodococcus erythropolis<\/jats:italic> R138 (<jats:italic>Re<\/jats:italic>BAL). While the overall sequence identity to the only experimentally confirmed benzaldehyde lyase from <jats:italic>Pseudomonas fluorescens<\/jats:italic> Biovar I (<jats:italic>Pf<\/jats:italic>BAL) was only 65\u2009%, comparison of a structural model of <jats:italic>Re<\/jats:italic>BAL with the crystal structure of <jats:italic>Pf<\/jats:italic>BAL revealed only four divergent amino acids in the substrate binding cavity. Based on rational design, we generated two <jats:italic>Re<\/jats:italic>BAL variants, which were characterized along with the wild\u2010type enzyme in terms of their substrate spectrum, thermostability and biocatalytic performance in the presence of different co\u2010solvents. We found that the new enzyme variants have a significantly higher thermostability (up to 22\u2009\u00b0C increase in T<jats:sub>50<\/jats:sub>) and a different co\u2010solvent\u2010dependent activity. Using the most stable variant immobilized in packed\u2010bed reactors via the SpyCatcher\/SpyTag system, (<jats:italic>R<\/jats:italic>)\u2010benzoin was synthesized from benzaldehyde over a period of seven days with a stable space\u2010time\u2010yield of 9.3\u2005mmol\u2009\u22c5\u2009L<jats:sup>\u20101<\/jats:sup>\u2009\u22c5\u2009d<jats:sup>\u22121<\/jats:sup>. Our work expands the important class of benzaldehyde lyases and therefore contributes to the development of continuous biocatalytic processes for the production of \u03b1\u2010hydroxy ketones and APIs.<\/jats:p>","DOI":"10.1002\/cbic.202100468","type":"journal-article","created":{"date-parts":[[2021,9,24]],"date-time":"2021-09-24T05:10:43Z","timestamp":1632460243000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Modeling\u2010Assisted Design of Thermostable Benzaldehyde Lyases from <i>Rhodococcus erythropolis<\/i> for Continuous Production of \u03b1\u2010Hydroxy Ketones"],"prefix":"10.1002","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2195-1927","authenticated-orcid":false,"given":"Martin","family":"Peng","sequence":"first","affiliation":[{"name":"Karlsruhe Institute of Technology (KIT) Institute for Biological Interfaces (IBG 1)  Hermann-von-Helmholtz-Platz\u20051 76344 Eggenstein-Leopoldshafen Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7010-8031","authenticated-orcid":false,"given":"Dominik L.","family":"Siebert","sequence":"additional","affiliation":[{"name":"Karlsruhe Institute of Technology (KIT) Institute for Biological Interfaces (IBG 1)  Hermann-von-Helmholtz-Platz\u20051 76344 Eggenstein-Leopoldshafen Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2174-2225","authenticated-orcid":false,"given":"Martin K. M.","family":"Engqvist","sequence":"additional","affiliation":[{"name":"Chalmers University of Technology Department of Biology and Biological Engineering Division of Systems and Synthetic Biology  Kemiv\u00e4gen 10 412 96 Gothenburg Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8837-081X","authenticated-orcid":false,"given":"Christof M.","family":"Niemeyer","sequence":"additional","affiliation":[{"name":"Karlsruhe Institute of Technology (KIT) Institute for Biological Interfaces (IBG 1)  Hermann-von-Helmholtz-Platz\u20051 76344 Eggenstein-Leopoldshafen Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7909-8191","authenticated-orcid":false,"given":"Kersten S.","family":"Rabe","sequence":"additional","affiliation":[{"name":"Karlsruhe Institute of Technology (KIT) Institute for Biological Interfaces (IBG 1)  Hermann-von-Helmholtz-Platz\u20051 76344 Eggenstein-Leopoldshafen Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2021,10,8]]},"reference":[{"key":"e_1_2_7_1_1","doi-asserted-by":"publisher","DOI":"10.1021\/ar900196n"},{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1002\/cctc.201100145"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bioorg.2021.104644"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1128\/jb.171.5.2401-2405.1989"},{"key":"e_1_2_7_5_1","unstructured":"\u00a0"},{"key":"e_1_2_7_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1119(94)90218-6"},{"key":"e_1_2_7_7_2","unstructured":"E. Janzen M. Pohl \u201cPseudomonas fluorescens biovar I benzaldehyde lyase (bznB) gene complete cds\u201d can be found underhttps:\/\/www.ncbi.nlm.nih.gov\/nuccore\/9965497 2000."},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkaa1025"},{"key":"e_1_2_7_9_1","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.201600462"},{"key":"e_1_2_7_10_1","doi-asserted-by":"publisher","DOI":"10.1002\/chem.201602084"},{"key":"e_1_2_7_11_1","unstructured":"\u00a0"},{"key":"e_1_2_7_12_2","doi-asserted-by":"publisher","DOI":"10.1002\/adsc.200700550"},{"key":"e_1_2_7_13_2","doi-asserted-by":"publisher","DOI":"10.1002\/chem.201406156"},{"key":"e_1_2_7_14_2","doi-asserted-by":"publisher","DOI":"10.1021\/ol034415b"},{"key":"e_1_2_7_15_2","doi-asserted-by":"publisher","DOI":"10.1002\/1615-4169(200201)344:1<96::AID-ADSC96>3.0.CO;2-Z"},{"key":"e_1_2_7_16_2","doi-asserted-by":"publisher","DOI":"10.1002\/cctc.201300093"},{"key":"e_1_2_7_17_2","doi-asserted-by":"publisher","DOI":"10.1039\/c0ob01121e"},{"key":"e_1_2_7_18_1","doi-asserted-by":"publisher","DOI":"10.1002\/adsc.201000735"},{"key":"e_1_2_7_19_1","doi-asserted-by":"publisher","DOI":"10.1002\/anie.201702278"},{"key":"e_1_2_7_19_2","doi-asserted-by":"publisher","DOI":"10.1002\/ange.201702278"},{"key":"e_1_2_7_20_1","doi-asserted-by":"publisher","DOI":"10.1002\/anie.202100534"},{"key":"e_1_2_7_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbapap.2005.08.025"},{"key":"e_1_2_7_22_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1500545112"},{"key":"e_1_2_7_23_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bioorg.2006.09.002"},{"key":"e_1_2_7_24_1","doi-asserted-by":"publisher","DOI":"10.1002\/bit.21189"},{"key":"e_1_2_7_25_1","doi-asserted-by":"publisher","DOI":"10.1021\/op0601316"},{"key":"e_1_2_7_26_1","doi-asserted-by":"publisher","DOI":"10.1002\/biot.200600030"},{"key":"e_1_2_7_27_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tetasy.2004.07.039"},{"key":"e_1_2_7_28_1","doi-asserted-by":"publisher","DOI":"10.1039\/b819722a"},{"key":"e_1_2_7_29_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbiotec.2016.12.004"},{"key":"e_1_2_7_30_1","unstructured":"\u00a0"},{"key":"e_1_2_7_31_2","doi-asserted-by":"publisher","DOI":"10.1039\/C8CY01891J"},{"key":"e_1_2_7_32_2","first-page":"1842","volume":"351","author":"J\u00e4ger V. D.","year":"2019","journal-title":"Adv. Synth. Catal."},{"key":"e_1_2_7_33_2","doi-asserted-by":"publisher","DOI":"10.1021\/acssynbio.8b00274"},{"key":"e_1_2_7_34_1","doi-asserted-by":"publisher","DOI":"10.1021\/acssynbio.9b00099"},{"key":"e_1_2_7_35_1","doi-asserted-by":"publisher","DOI":"10.1093\/protein\/gzn001"},{"key":"e_1_2_7_36_1","first-page":"478925","author":"Natalia D.","year":"2011","journal-title":"Biotechnol. Res. Int."},{"key":"e_1_2_7_37_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00253-016-7809-9"},{"key":"e_1_2_7_38_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10709-015-9827-4"},{"key":"e_1_2_7_39_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi8004413"},{"key":"e_1_2_7_40_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gky427"},{"key":"e_1_2_7_41_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-09654-8"},{"key":"e_1_2_7_42_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkw1132"},{"key":"e_1_2_7_43_1","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btq662"},{"key":"e_1_2_7_44_1","doi-asserted-by":"publisher","DOI":"10.1002\/elps.200900140"},{"key":"e_1_2_7_45_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1742-4658.2005.04998.x"},{"key":"e_1_2_7_46_1","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.200600277"},{"key":"e_1_2_7_47_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gky473"},{"key":"e_1_2_7_48_1","unstructured":"\u00a0"},{"key":"e_1_2_7_49_2","doi-asserted-by":"publisher","DOI":"10.1021\/acscatal.9b05223"},{"key":"e_1_2_7_50_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00792-011-0363-6"},{"key":"e_1_2_7_51_2","doi-asserted-by":"publisher","DOI":"10.1042\/bj3120527"},{"key":"e_1_2_7_52_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcatb.2009.02.021"},{"key":"e_1_2_7_53_1","unstructured":"\u00a0"},{"key":"e_1_2_7_54_2","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2007.354"},{"key":"e_1_2_7_55_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja00783a034"},{"key":"e_1_2_7_56_1","unstructured":"\u00a0"},{"key":"e_1_2_7_57_2","doi-asserted-by":"publisher","DOI":"10.1039\/C5GC00342C"},{"key":"e_1_2_7_58_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcatb.2014.06.003"},{"key":"e_1_2_7_59_1","unstructured":"\u00a0"},{"key":"e_1_2_7_60_2","author":"Schweiger A. K.","year":"2019","journal-title":"ACS Sustainable Chem. Eng."},{"key":"e_1_2_7_61_2","doi-asserted-by":"publisher","DOI":"10.3390\/catal10040447"},{"key":"e_1_2_7_62_2","doi-asserted-by":"publisher","DOI":"10.3390\/molecules24040792"},{"key":"e_1_2_7_63_1","doi-asserted-by":"publisher","DOI":"10.1002\/adsc.201200284"},{"key":"e_1_2_7_64_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bioorg.2009.03.001"},{"key":"e_1_2_7_65_1","unstructured":"\u00a0"},{"key":"e_1_2_7_66_2","doi-asserted-by":"publisher","DOI":"10.1002\/adsc.200700230"},{"key":"e_1_2_7_67_2","doi-asserted-by":"publisher","DOI":"10.1039\/D1GC00561H"},{"key":"e_1_2_7_68_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1115485109"},{"key":"e_1_2_7_69_1","unstructured":"\u00a0"},{"key":"e_1_2_7_70_2","doi-asserted-by":"publisher","DOI":"10.1039\/D0SC01878C"},{"key":"e_1_2_7_71_2","doi-asserted-by":"publisher","DOI":"10.1021\/acscatal.8b00986"},{"key":"e_1_2_7_72_2","doi-asserted-by":"publisher","DOI":"10.1002\/anie.201810331"},{"key":"e_1_2_7_72_3","doi-asserted-by":"publisher","DOI":"10.1002\/ange.201810331"},{"key":"e_1_2_7_73_2","doi-asserted-by":"publisher","DOI":"10.3390\/mi10110783"},{"key":"e_1_2_7_74_2","doi-asserted-by":"publisher","DOI":"10.3390\/mi10120795"},{"key":"e_1_2_7_75_2","doi-asserted-by":"publisher","DOI":"10.1021\/acsanm.9b01975"},{"key":"e_1_2_7_76_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1401291111"},{"key":"e_1_2_7_77_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.chembiol.2021.01.011"},{"key":"e_1_2_7_78_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cbpa.2019.11.008"},{"key":"e_1_2_7_79_2","doi-asserted-by":"crossref","first-page":"409","DOI":"10.1002\/9783527817290.ch12","volume-title":"Catalyst Immobilization: Methods and Applications","author":"Romero-Fern\u00e1ndez M.","year":"2020"},{"key":"e_1_2_7_80_1","doi-asserted-by":"publisher","DOI":"10.1021\/acscatal.7b02230"},{"key":"e_1_2_7_81_1","doi-asserted-by":"publisher","DOI":"10.1016\/0167-7799(89)90035-8"},{"key":"e_1_2_7_82_1","doi-asserted-by":"publisher","DOI":"10.1002\/anie.201609590"},{"key":"e_1_2_7_82_2","doi-asserted-by":"publisher","DOI":"10.1002\/ange.201609590"},{"key":"e_1_2_7_83_1","doi-asserted-by":"publisher","DOI":"10.1002\/cctc.202000145"},{"key":"e_1_2_7_84_1","doi-asserted-by":"publisher","DOI":"10.1039\/C9SC04074A"},{"key":"e_1_2_7_85_1","unstructured":"\u00a0"},{"key":"e_1_2_7_86_2","doi-asserted-by":"publisher","DOI":"10.1002\/prot.21227"},{"key":"e_1_2_7_87_2","doi-asserted-by":"publisher","DOI":"10.1006\/jmbi.2001.5226"}],"container-title":["ChemBioChem"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cbic.202100468","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/cbic.202100468","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/chemistry-europe.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cbic.202100468","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T20:46:50Z","timestamp":1760302010000},"score":1,"resource":{"primary":{"URL":"https:\/\/chemistry-europe.onlinelibrary.wiley.com\/doi\/10.1002\/cbic.202100468"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,10,8]]},"references-count":90,"journal-issue":{"issue":"7","published-print":{"date-parts":[[2022,4,5]]}},"alternative-id":["10.1002\/cbic.202100468"],"URL":"https:\/\/doi.org\/10.1002\/cbic.202100468","archive":["Portico"],"relation":{},"ISSN":["1439-4227","1439-7633"],"issn-type":[{"value":"1439-4227","type":"print"},{"value":"1439-7633","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,10,8]]},"assertion":[{"value":"2021-09-02","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2021-10-08","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e202100468"}}