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Dye\u2010coupled assays showed that the earlier Lns exhibit a higher enzyme activity than the late Lns. Despite widespread use, there are limitations: oftentimes a pH of 9 and activators are required for the assay. Moreover, Ln\u2010MDH variants are not obtained by isolation from the cells grown with the respective Ln, but by incubation of an apo\u2010MDH with the Ln. Herein, we report the cultivation of Ln\u2010dependent methanotroph <jats:italic>Methylacidiphilum fumariolicum<\/jats:italic> SolV with nine different Lns, the isolation of the respective MDHs and the assessment of the enzyme activity using the dye\u2010coupled assay. We compare these results with a protein\u2010coupled assay using its physiological electron acceptor cytochrome <jats:italic>c<jats:sub>GJ<\/jats:sub><\/jats:italic> (cyt <jats:italic>c<jats:sub>GJ<\/jats:sub><\/jats:italic>). Depending on the assay, two distinct trends are observed among the Ln series. The specific enzyme activity of La\u2010, Ce\u2010 and Pr\u2010MDH, as measured by the protein\u2010coupled assay, exceeds that measured by the dye\u2010coupled assay. This suggests that early Lns also have a positive effect on the interaction between XoxF\u2010MDH and its cyt <jats:italic>c<jats:sub>GJ<\/jats:sub><\/jats:italic> thereby increasing functional efficiency.<\/jats:p>","DOI":"10.1002\/cbic.202300811","type":"journal-article","created":{"date-parts":[[2024,1,25]],"date-time":"2024-01-25T05:30:58Z","timestamp":1706160658000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Assessing Lanthanide\u2010Dependent Methanol Dehydrogenase Activity: The Assay Matters"],"prefix":"10.1002","volume":"25","author":[{"given":"Manh Tri","family":"Phi","sequence":"first","affiliation":[{"name":"Department of Chemistry Ludwig-Maximilians-Universit\u00e4t M\u00fcnchen  Butenandtstr. 5\u201313 81377 M\u00fcnchen Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Helena","family":"Singer","sequence":"additional","affiliation":[{"name":"Department of Chemistry Ludwig-Maximilians-Universit\u00e4t M\u00fcnchen  Butenandtstr. 5\u201313 81377 M\u00fcnchen Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Felix","family":"Z\u00e4h","sequence":"additional","affiliation":[{"name":"Department of Chemistry Ludwig-Maximilians-Universit\u00e4t M\u00fcnchen  Butenandtstr. 5\u201313 81377 M\u00fcnchen Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christoph","family":"Haisch","sequence":"additional","affiliation":[{"name":"Faculty of Chemistry Technical University of Munich  Lichtenbergstr. 4 85748 Garching Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sabine","family":"Schneider","sequence":"additional","affiliation":[{"name":"Department of Chemistry Ludwig-Maximilians-Universit\u00e4t M\u00fcnchen  Butenandtstr. 5\u201313 81377 M\u00fcnchen Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1990-9030","authenticated-orcid":false,"given":"Huub J. M.","family":"Op den Camp","sequence":"additional","affiliation":[{"name":"Department of Microbiology Research Institute for Biological and Environmental Sciences Radboud University Nijmegen  Heyendaalseweg 135 6525 AJ Nijmegen The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2197-136X","authenticated-orcid":false,"given":"Lena J.","family":"Daumann","sequence":"additional","affiliation":[{"name":"Department of Chemistry Ludwig-Maximilians-Universit\u00e4t M\u00fcnchen  Butenandtstr. 5\u201313 81377 M\u00fcnchen Germany"},{"name":"Chair of Bioinorganic Chemistry Heinrich-Heine-Universit\u00e4t D\u00fcsseldorf  Universit\u00e4tsstra\u00dfe 1 40225 D\u00fcsseldorf Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2024,1,25]]},"reference":[{"key":"e_1_2_8_1_1","unstructured":"\u00a0"},{"key":"e_1_2_8_2_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0050480"},{"key":"e_1_2_8_3_2","volume":"35","author":"Kato S.","year":"2020","journal-title":"Microbes Environ."},{"key":"e_1_2_8_4_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00248-016-0808-7"},{"key":"e_1_2_8_5_2","doi-asserted-by":"publisher","DOI":"10.1111\/1462-2920.12896"},{"key":"e_1_2_8_6_2","doi-asserted-by":"publisher","DOI":"10.1111\/1462-2920.14062"},{"key":"e_1_2_8_7_2","volume":"97","author":"Roszczenko-Jasi\u0144ska P.","year":"2020","journal-title":"FEMS Microbiol. 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