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doi:10.22028/D291-33430
Titel: | Identification and circumvention of bottlenecks in CYP21A2-mediated premedrol production using recombinant Escherichia coli |
VerfasserIn: | König, Lisa Brixius-Anderko, Simone Milhim, Mohammed Tavouli-Abbas, Daniela Hutter, Michael C. Hannemann, Frank Bernhardt, Rita |
Sprache: | Englisch |
Titel: | Biotechnology and Bioengineering |
Bandnummer: | 117 |
Heft: | 4 |
Seiten: | 901–911 |
Verlag/Plattform: | Wiley |
Erscheinungsjahr: | 2019 |
Freie Schlagwörter: | C21 hydroxylation CYP21A2 cytochrome b5 enzyme engineering whole‐cell biotransformation |
DDC-Sachgruppe: | 570 Biowissenschaften, Biologie |
Dokumenttyp: | Journalartikel / Zeitschriftenartikel |
Abstract: | Synthetic glucocorticoids such as methylprednisolone are compounds of fundamental interest to the pharmaceutical industry as their modifications within the sterane scaffold lead to higher inflammatory potency and reduced side effects compared with their parent compound cortisol. In methylprednisolone production, the complex chemical hydroxylation of its precursor medrane in position C21 exhibits poor stereo‐ and regioselectivity making the process unprofitable and unsustainable. By contrast, the use of a recombinant E. coli system has recently shown high suitability and efficiency. In this study, we aim to overcome limitations in this biotechnological medrane conversion yielding the essential methylprednisolone‐precursor premedrol by optimizing the CYP21A2‐based whole‐cell system on a laboratory scale. We successfully improved the whole‐cell process in terms of premedrol production by (a) improving the electron supply to CYP21A2; here we use the N‐terminally truncated version of the bovine NADPH‐dependent cytochrome P450 reductase (bCPR−27) and coexpression of microsomal cytochrome b5; (b) enhancing substrate access to the heme by modification of the CYP21A2 substrate access channel; and (c) circumventing substrate inhibition which is presumed to be the main limiting factor of the presented system by developing an improved fed‐batch protocol. By overcoming the presented limitations in whole‐cell biotransformation, we were able to achieve a more than 100% improvement over the next best system under equal conditions resulting in 691 mg·L−1·d−1 premedrol. |
DOI der Erstveröffentlichung: | 10.1002/bit.27246 |
Link zu diesem Datensatz: | urn:nbn:de:bsz:291--ds-334306 hdl:20.500.11880/30741 http://dx.doi.org/10.22028/D291-33430 |
ISSN: | 1097-0290 0006-3592 |
Datum des Eintrags: | 26-Feb-2021 |
Bezeichnung des in Beziehung stehenden Objekts: | Supporting Information |
In Beziehung stehendes Objekt: | https://onlinelibrary.wiley.com/action/downloadSupplement?doi=10.1002%2Fbit.27246&file=bit27246-sup-0001-Supplementary.docx |
Fakultät: | NT - Naturwissenschaftlich- Technische Fakultät |
Fachrichtung: | NT - Biowissenschaften |
Professur: | NT - Prof. Dr. Volkhard Helms NT - Prof. Dr. Bruce Morgan NT - Keiner Professur zugeordnet |
Sammlung: | SciDok - Der Wissenschaftsserver der Universität des Saarlandes |
Dateien zu diesem Datensatz:
Datei | Beschreibung | Größe | Format | |
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bit.27246.pdf | 1,47 MB | Adobe PDF | Öffnen/Anzeigen |
Diese Ressource wurde unter folgender Copyright-Bestimmung veröffentlicht: Lizenz von Creative Commons