{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,26]],"date-time":"2026-08-26T23:07:15Z","timestamp":1787785635698,"version":"build-2784847793"},"reference-count":20,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2019,1,8]],"date-time":"2019-01-08T00:00:00Z","timestamp":1546905600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001446","name":"Economic Development Board - Singapore","doi-asserted-by":"publisher","award":["279\u2010000\u2010348\u2010592"],"award-info":[{"award-number":["279\u2010000\u2010348\u2010592"]}],"id":[{"id":"10.13039\/501100001446","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100004330","name":"GlaxoSmithKline","doi-asserted-by":"publisher","award":["279\u2010000\u2010348\u2010592"],"award-info":[{"award-number":["279\u2010000\u2010348\u2010592"]}],"id":[{"id":"10.13039\/100004330","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Biotech &amp; Bioengineering"],"published-print":{"date-parts":[[2019,3]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Alcohol dehydrogenase (ADH) and amine dehydrogenase (AmDH)\u2010catalyzed one\u2010pot cascade conversion of an alcohol to an amine provides a simple preparation of chiral amines. To enhance the cofactor recycling in this reaction, we report a new concept of coupling whole\u2010cells with the cell\u2010free system to enable separated intracellular and extracellular cofactor regeneration and recycling. This was demonstrated by the respective biotransformation of racemic 4\u2010phenyl\u20102\u2010butanol <jats:bold>1a<\/jats:bold> and 1\u2010phenyl\u20102\u2010propanol <jats:bold>1b<\/jats:bold> to (<jats:italic>R<\/jats:italic>)\u20104\u2010phenylbutan\u20102\u2010amine <jats:bold>3a<\/jats:bold> and (<jats:italic>R<\/jats:italic>)\u20101\u2010phenylpropan\u20102\u2010amine <jats:bold>3b<\/jats:bold>. <jats:italic>Escherichia coli<\/jats:italic> cells expressing <jats:italic>S<\/jats:italic>\u2010enantioselective CpsADH, <jats:italic>R<\/jats:italic>\u2010enantioselective PfODH, and NADH oxidase (NOX) was developed to oxidize racemic alcohols <jats:bold>1a\u2013b<\/jats:bold> to ketones <jats:bold>2a\u2013b<\/jats:bold> with full conversion via intracellular NAD<jats:sup>+<\/jats:sup> recycling. AmDH and glucose dehydrogenase (GDH) were used to convert ketones <jats:bold>2a\u2013b<\/jats:bold> to amines (<jats:italic>R<\/jats:italic>)\u2010<jats:bold>3a\u2013b<\/jats:bold> with 89\u201394% conversion and 891\u2013943 times recycling of NADH. Combining the cells and enzymes for the cascade transformation of racemic alcohols <jats:bold>1a\u2013b<\/jats:bold> gave 70% and 48% conversion to the amines (<jats:italic>R<\/jats:italic>)\u2010<jats:bold>3a<\/jats:bold> and (<jats:italic>R<\/jats:italic>)\u20103<jats:bold>b<\/jats:bold> in 99% <jats:italic>ee<\/jats:italic>, with a\u00a0total turnover number (TTN) of 350 and 240 for NADH recycling, respectively. Improved results were obtained by using the <jats:italic>E. coli<\/jats:italic> cells with immobilized AmDH and GDH: (<jats:italic>R<\/jats:italic>)\u2010<jats:bold>3a<\/jats:bold> was produced in 99% <jats:italic>ee<\/jats:italic> with 71\u201384% conversion and a TTN of 1410\u20101260 for NADH recycling, the highest value so far for the ADH\u2013AmDH\u2010catalyzed cascade conversion of alcohols to amines. The concept might be generally applicable to this type of reactions.<\/jats:p>","DOI":"10.1002\/bit.26896","type":"journal-article","created":{"date-parts":[[2018,12,8]],"date-time":"2018-12-08T07:39:06Z","timestamp":1544254746000},"page":"536-542","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":34,"title":["Enhancing cofactor recycling in the bioconversion of racemic alcohols to chiral amines with alcohol dehydrogenase and amine dehydrogenase by coupling cells and cell\u2010free system"],"prefix":"10.1002","volume":"116","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2528-2839","authenticated-orcid":false,"given":"Ji","family":"Liu","sequence":"first","affiliation":[{"name":"Department of Chemical &amp; Biomolecular Engineering National University of Singapore, 4 Engineering Drive 4  Singapore"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7370-2562","authenticated-orcid":false,"given":"Zhi","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Chemical &amp; Biomolecular Engineering National University of Singapore, 4 Engineering Drive 4  Singapore"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2019,1,8]]},"reference":[{"key":"e_1_2_6_1_2_1","doi-asserted-by":"publisher","DOI":"10.1002\/anie.201107813"},{"key":"e_1_2_6_1_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/nchem.2782"},{"key":"e_1_2_6_1_4_1","doi-asserted-by":"publisher","DOI":"10.1002\/cctc.201701366"},{"key":"e_1_2_6_1_5_1","doi-asserted-by":"publisher","DOI":"10.1002\/cctc.201500785"},{"key":"e_1_2_6_1_6_1","doi-asserted-by":"publisher","DOI":"10.1039\/B703488C"},{"key":"e_1_2_6_1_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cbpa.2017.09.008"},{"key":"e_1_2_6_1_8_1","unstructured":"Hummel W. &Riebel B.(1999). 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(R)\u2010specific alcohol dehydrogenases from lactobacillus with improved catalytic activity using an NAD+substrate."},{"key":"e_1_2_6_1_9_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi051596b"},{"key":"e_1_2_6_1_10_1","doi-asserted-by":"publisher","DOI":"10.1039\/C7OB01927K"},{"key":"e_1_2_6_1_11_1","doi-asserted-by":"publisher","DOI":"10.1021\/op4000237"},{"key":"e_1_2_6_1_12_1","doi-asserted-by":"publisher","DOI":"10.1002\/cctc.201601446"},{"key":"e_1_2_6_1_13_1","doi-asserted-by":"publisher","DOI":"10.1002\/ange.201705848"},{"key":"e_1_2_6_1_14_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.aac9283"},{"key":"e_1_2_6_1_15_1","doi-asserted-by":"publisher","DOI":"10.1002\/cctc.201701092"},{"key":"e_1_2_6_1_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.copbio.2016.05.005"},{"key":"e_1_2_6_1_17_1","doi-asserted-by":"publisher","DOI":"10.1002\/cctc.201701669"},{"key":"e_1_2_6_1_18_1","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms11917"},{"key":"e_1_2_6_1_19_1","doi-asserted-by":"publisher","DOI":"10.1002\/anie.201311091"},{"key":"e_1_2_6_1_20_1","doi-asserted-by":"publisher","DOI":"10.1021\/cs501906r"},{"key":"e_1_2_6_1_21_1","doi-asserted-by":"publisher","DOI":"10.1021\/cs5016113"}],"container-title":["Biotechnology and Bioengineering"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/bit.26896","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/bit.26896","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/bit.26896","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T22:13:28Z","timestamp":1760048008000},"score":1,"resource":{"primary":{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/10.1002\/bit.26896"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,1,8]]},"references-count":20,"journal-issue":{"issue":"3","published-print":{"date-parts":[[2019,3]]}},"alternative-id":["10.1002\/bit.26896"],"URL":"https:\/\/doi.org\/10.1002\/bit.26896","archive":["Portico"],"relation":{},"ISSN":["0006-3592","1097-0290"],"issn-type":[{"value":"0006-3592","type":"print"},{"value":"1097-0290","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,1,8]]},"assertion":[{"value":"2018-10-02","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2018-12-06","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2019-01-08","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}