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A regulated synthetic operon facilitates stable overexpression of multigene terpenoid pathway in Bacillus subtilis
Abdallah I.I.a, Xue D.a, Pramastya H.a,b, van Merkerk R.a, Setroikromo R.a, Quax W.J.a
a Department of Chemical and Pharmaceutical Biology, Groningen Research Institute of Pharmacy, University of Groningen, Groningen, 9713 AV, Netherlands
b Pharmaceutical Biology Research Group, School of Pharmacy, Institut Teknologi Bandung, Bandung, 40132, Indonesia
[vc_row][vc_column][vc_row_inner][vc_column_inner][vc_separator css=”.vc_custom_1624529070653{padding-top: 30px !important;padding-bottom: 30px !important;}”][/vc_column_inner][/vc_row_inner][vc_row_inner layout=”boxed”][vc_column_inner width=”3/4″ css=”.vc_custom_1624695412187{border-right-width: 1px !important;border-right-color: #dddddd !important;border-right-style: solid !important;border-radius: 1px !important;}”][vc_empty_space][megatron_heading title=”Abstract” size=”size-sm” text_align=”text-left”][vc_column_text]© 2020, Society for Industrial Microbiology and Biotechnology.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 C30 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 C30 carotenoid production.[/vc_column_text][vc_empty_space][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][vc_empty_space][megatron_heading title=”Author keywords” size=”size-sm” text_align=”text-left”][vc_column_text]Bacillus subtilis,Biosynthetic Pathways,DNA Replication,Genetic Vectors,Multigene Family,Operon,Plasmids,Terpenes[/vc_column_text][vc_empty_space][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][vc_empty_space][megatron_heading title=”Indexed keywords” size=”size-sm” text_align=”text-left”][vc_column_text]Bacillus subtilis,Carotenoids,Cell factory,MEP,Stability[/vc_column_text][vc_empty_space][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][vc_empty_space][megatron_heading title=”Funding details” size=”size-sm” text_align=”text-left”][vc_column_text][{‘$’: ‘We thank I. Maeda for providing the pHYcrtMN plasmid. Funding for this work was obtained through EuroCoRes SYNBIO (SYNMET), NWO-ALW 855.01.161, EU FP-7 grant 289540 (PROMYSE). IIA is a recipient of Erasmus Mundus Action 2, Strand 1, Fatima Al Fihri project ALFI1200161 scholarship and is on study leave from Faculty of Pharmacy, Alexandria University. HP is a recipient of Bernoulli scholarship from University of Groningen.’}, {‘$’: ‘We thank I. Maeda for providing the pHYcrtMN plasmid. Funding for this work was obtained through EuroCoRes SYNBIO (SYNMET), NWO-ALW 855.01.161, EU FP-7 grant 289540 (PROMYSE). IIA is a recipient of Erasmus Mundus Action 2, Strand 1, Fatima Al Fihri project ALFI1200161 scholarship and is on study leave from Faculty of Pharmacy, Alexandria University. HP is a recipient of Bernoulli scholarship from University of Groningen.’}][/vc_column_text][vc_empty_space][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][vc_empty_space][megatron_heading title=”DOI” size=”size-sm” text_align=”text-left”][vc_column_text]https://doi.org/10.1007/s10295-019-02257-4[/vc_column_text][/vc_column_inner][vc_column_inner width=”1/4″][vc_column_text]Widget Plumx[/vc_column_text][/vc_column_inner][/vc_row_inner][/vc_column][/vc_row][vc_row][vc_column][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][/vc_column][/vc_row]