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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

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