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A regulated synthetic operon facilitates stable overexpression of multigene terpenoid pathway in Bacillus subtilis

Abdallah, I. I., Xue, D., Pramastya, H., van Merkerk, R., Setroikromo, R. & Quax, W. J., Feb-2020, In : Journal of Industrial Microbiology & Biotechnology. 47, 2, p. 243-249 7 p.

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  • A regulated synthetic operon facilitates stable overexpression of multigene terpenoid pathway in Bacillus subtilis

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DOI

The creation of microbial cell factories for sustainable production of natural products is important for medical and industrial applications. This requires stable expression of biosynthetic pathways in a host organism with favorable fermentation properties such as Bacillus subtilis. The aim of this study is to construct B. subtilis strains that produce valuable terpenoid compounds by overexpressing the innate methylerythritol phosphate (MEP) pathway. A synthetic operon allowing the concerted and regulated expression of multiple genes was developed. Up to 8 genes have been combined in this operon and a stably inherited plasmid-based vector was constructed resulting in a high production of C-30 carotenoids. For this, two vectors were examined, one with rolling circle replication and another with theta replication. Theta-replication constructs were clearly superior in structural and segregational stability compared to rolling circle constructs. A strain overexpressing all eight genes of the MEP pathway on a theta-replicating plasmid clearly produced the highest level of carotenoids. The level of transcription for each gene in the operon was similar as RT-qPCR analysis indicated. Hence, that corresponding strain can be used as a stable cell factory for production of terpenoids. This is the first report of merging and stably expressing this large-size operon (eight genes) from a plasmid-based system in B. subtilis enabling high C-30 carotenoid production.

Original languageEnglish
Pages (from-to)243-249
Number of pages7
JournalJournal of Industrial Microbiology & Biotechnology
Volume47
Issue number2
Publication statusPublished - Feb-2020

    Keywords

  • Bacillus subtilis, MEP, Carotenoids, Cell factory, Stability, ESCHERICHIA-COLI, RIBOFLAVIN PRODUCTION, EXPRESSION VECTORS, PLASMID PLS32, CONSTRUCTION, REPLICATION, OPTIMIZATION, CLONING, OVERPRODUCTION, MOLASSES

ID: 120758715