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In this study, an <jats:italic>Acidovorax\u2010<\/jats:italic>derived <jats:italic>CHMO<\/jats:italic> gene was expressed in <jats:italic>Pseudomonas taiwanensis<\/jats:italic> VLB120. Upon purification, the enzyme was characterized in vitro and shown to feature a broad substrate spectrum and up to 100% conversion in 6\u2009h. Furthermore, we determined and compared the cyclohexanone conversion kinetics for different CHMO\u2010biocatalyst formats, that is, isolated enzyme, suspended whole cells, and biofilms, the latter two based on recombinant CHMO\u2010containing <jats:italic>P. taiwanensis<\/jats:italic> VLB120. Biofilms showed less favorable values for <jats:italic>K<\/jats:italic><jats:sub>S<\/jats:sub> (9.3\u2010fold higher) and <jats:italic>k<\/jats:italic><jats:sub>cat<\/jats:sub> (4.8\u2010fold lower) compared with corresponding <jats:italic>K<\/jats:italic><jats:sub>M<\/jats:sub> and <jats:italic>k<\/jats:italic><jats:sub>cat<\/jats:sub> values of isolated CHMO, but a favorable <jats:italic>K<\/jats:italic><jats:sub>I<\/jats:sub> for cyclohexanone (5.3\u2010fold higher). The unfavorable <jats:italic>K<\/jats:italic><jats:sub>S<\/jats:sub> and <jats:italic>k<\/jats:italic><jats:sub>cat<\/jats:sub> values are related to mass transfer\u2010 and possibly heterogeneity issues and deserve further investigation and engineering, to exploit the high potential of biofilms regarding process stability. Suspended cells showed only 1.8\u2010fold higher <jats:italic>K<\/jats:italic><jats:sub>S<\/jats:sub>, but 1.3\u2010 and 4.2\u2010fold higher <jats:italic>k<\/jats:italic><jats:sub>cat<\/jats:sub> and <jats:italic>K<\/jats:italic><jats:sub>I<\/jats:sub> values than isolated CHMO. This together with the efficient NADPH regeneration via glucose metabolism makes this format highly promising from a kinetics perspective.<\/jats:p>","DOI":"10.1002\/bit.27791","type":"journal-article","created":{"date-parts":[[2021,4,12]],"date-time":"2021-04-12T08:03:57Z","timestamp":1618214637000},"page":"2719-2733","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Characterization of different biocatalyst formats for BVMO\u2010catalyzed cyclohexanone oxidation"],"prefix":"10.1002","volume":"118","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9149-0265","authenticated-orcid":false,"given":"Lisa","family":"Bretschneider","sequence":"first","affiliation":[{"name":"Department of Solar Materials Helmholtz\u2010Centre for Environmental Research \u2010 UFZ Leipzig 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