{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,30]],"date-time":"2026-06-30T22:00:54Z","timestamp":1782856854614,"version":"3.54.5"},"reference-count":44,"publisher":"Walter de Gruyter GmbH","issue":"11","funder":[{"name":"Arbeitsgemeinschaft industrieller Forschungsvereinigungen","award":["F 519F"],"award-info":[{"award-number":["F 519F"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2020,10,25]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Cytochrome P450s are an important group of enzymes catalyzing hydroxylation, and epoxidations reactions. In this work we describe the characterization of the CinA\u2013CinC fusion enzyme system of a previously reported P450 using genetically fused heme (CinA) and FMN (CinC) enzyme domains from<jats:italic>Citrobacter braaki<\/jats:italic>. We observed that mixing individually inactivated heme (-) with FMN (-) domain in the CinA-10aa linker - CinC fusion constructs results in recovered activity and the formation of (2S)-2\u03b2-hydroxy,1,8-cineole (174\u00a0\u00b5M), a similar amount when compared to the fully functional fusion protein (176\u00a0\u00b5M). We also studied the effect of the fusion linker length in the activity complementation assay. Our results suggests an intermolecular interaction between heme and FMN parts from different CinA\u2013CinC fusion protein similar to proposed mechanisms for P450 BM3 on the other hand, linker length plays a crucial influence on the activity of the fusion constructs. However, complementation assays show that inactive constructs with shorter linker lengths have functional subunits, and that the lack of activity might be due to incorrect interaction between fused enzymes.<\/jats:p>","DOI":"10.1515\/hsz-2020-0134","type":"journal-article","created":{"date-parts":[[2020,6,6]],"date-time":"2020-06-06T07:44:19Z","timestamp":1591429459000},"page":"1249-1255","source":"Crossref","is-referenced-by-count":3,"title":["Insights on intermolecular FMN-heme domain interaction and the role of linker length in cytochrome P450cin fusion proteins"],"prefix":"10.1515","volume":"401","author":[{"given":"Ketaki D.","family":"Belsare","sequence":"first","affiliation":[{"name":"Lehrstuhl f\u00fcr Biotechnologie, RWTH Aachen University, Worringerweg 3, Aachen, D-52074, Germany"},{"name":"University of California at San Francisco, 555 Mission Bay Blvd South, San Francisco, 94158, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna Jo\u00eblle","family":"Ruff","sequence":"additional","affiliation":[{"name":"Lehrstuhl f\u00fcr Biotechnologie, RWTH Aachen University, Worringerweg 3, Aachen, D-52074, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ronny","family":"Martinez","sequence":"additional","affiliation":[{"name":"Lehrstuhl f\u00fcr Biotechnologie, RWTH Aachen University, Worringerweg 3, Aachen, D-52074, Germany"},{"name":"Departamento de Ingenier\u00eda en Alimentos, Universidad de La Serena, Av. Ra\u00fal Bitr\u00e1n 1305, La Serena, 1720010, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ulrich","family":"Schwaneberg","sequence":"additional","affiliation":[{"name":"Lehrstuhl f\u00fcr Biotechnologie, RWTH Aachen University, Worringerweg 3, Aachen, D-52074, Germany"},{"name":"DWI \u2013\u00a0Leibniz Institut f\u00fcr Interaktive Materialien, Forckenbeckstra\u00dfe 50, Aachen, D-52074, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"374","reference":[{"key":"ref381","first-page":"1","article-title":"Fusion proteins of an enoate reductase and a Baeyer-Villiger monooxygenase faciltate the synthesis of a chiral lactones","volume":"398","year":"2017","journal-title":"Biol. Chem."},{"key":"ref161","first-page":"1","article-title":"Fusion proteins of an enoate reductase and a Baeyer-Villiger monooxygenase faciltate the synthesis of a chiral lactones","volume":"398","year":"2017","journal-title":"Biol. 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