A straightforward approach to synthesize 7-aminocephalosporanic acid in vivo in the cephalosporin C producer \(\textit {Acremonium chrysogenum}\)
- The pharmaceutical industry has developed various highly effective semi-synthetic cephalosporins, which are generated by modifying the side chains of the core molecule 7-aminocephalosporanic acid (7-ACA). In industrial productions, the 7-ACA nucleus is obtained in vitro from cephalosporin C (CPC) by chemical or enzymatic processes, which are waste intensive and associated with high production costs. Here, we used a transgenic in vivo approach to express bacterial genes for cephalosporin C acylase (CCA) in the CPC producer Acremonium chrysogenum. Western blot and mass spectrometry analyses verified that the heterologous enzymes are processed into α- and β-subunits in the fungal cell. Extensive HPLC analysis detected substrates and products of CCAs in both fungal mycelia and culture supernatants, with the highest amount of 7-ACA found in the latter. Using different incubation times, temperatures, and pH values, we explored the optimal conditions for the active bacterial acylase to convert CPC into 7-ACA in the culture supernatant. We calculated that the best transgenic fungal strains exhibit a one-step conversion rate of the bacterial acylase of 30%. Our findings can be considered a remarkable contribution to supporting future pharmaceutical manufacturing processes with reduced production costs.
Author: | Xuemei LinGND, Jan LambertzGND, Tim A. DahlmannORCiDGND, Marc M. NowaczykORCiDGND, Burghard KönigGND, Ulrich KückORCiDGND |
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URN: | urn:nbn:de:hbz:294-101891 |
DOI: | https://doi.org/10.3390/jof8050450 |
Parent Title (English): | Journal of fungi |
Publisher: | MDPI |
Place of publication: | Basel |
Document Type: | Article |
Language: | English |
Date of Publication (online): | 2023/09/26 |
Date of first Publication: | 2022/04/26 |
Publishing Institution: | Ruhr-Universität Bochum, Universitätsbibliothek |
Tag: | Open Access Fonds 7-Aminocephalosporanic acid; Acremonium chrysogenum; cephalosporin C acylase |
Volume: | 8 |
Issue: | 5, Artikel 450 |
First Page: | 450-1 |
Last Page: | 450-20 |
Note: | Article Processing Charge funded by the Deutsche Forschungsgemeinschaft (DFG) and the Open Access Publication Fund of Ruhr-Universität Bochum. |
Institutes/Facilities: | Fakultät für Biologie und Biotechnologie, Arbeitskreis Allgemeine und Molekulare Botanik |
Dewey Decimal Classification: | Naturwissenschaften und Mathematik / Biowissenschaften, Biologie, Biochemie |
open_access (DINI-Set): | open_access |
faculties: | Fakultät für Biologie und Biotechnologie |
Licence (English): | Creative Commons - CC BY 4.0 - Attribution 4.0 International |