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Cycling between gro...
Cycling between growth and production phases increases cyanobacteria bioproduction of lactate
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- Shabestary, Kiyan (författare)
- KTH,Systembiologi,Science for Life Laboratory, SciLifeLab
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- Hernandez, Hugo Pineda (författare)
- Univ Amsterdam, Fac Sci, Swammerdam Inst Life Sci, Mol Microbial Physiol Grp, Sci Pk 904, NL-1098 XH Amsterdam, Netherlands.
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- Miao, Rui (författare)
- KTH,Science for Life Laboratory, SciLifeLab,Systembiologi
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- Ljungqvist, Emil E. (författare)
- KTH,Industriell bioteknologi,Science for Life Laboratory, SciLifeLab
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- Hallman, Olivia (författare)
- KTH,Skolan för kemi, bioteknologi och hälsa (CBH),Science for Life Laboratory, SciLifeLab
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- Sporre, Emil (författare)
- KTH,Science for Life Laboratory, SciLifeLab,Systembiologi
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- dos Santos, Filipe Branco (författare)
- Univ Amsterdam, Fac Sci, Swammerdam Inst Life Sci, Mol Microbial Physiol Grp, Sci Pk 904, NL-1098 XH Amsterdam, Netherlands.
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- Hudson, Elton P. (författare)
- KTH,Science for Life Laboratory, SciLifeLab,Systembiologi
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(creator_code:org_t)
- Elsevier BV, 2021
- 2021
- Engelska.
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Ingår i: Metabolic engineering. - : Elsevier BV. - 1096-7176 .- 1096-7184. ; 68, s. 131-141
- Relaterad länk:
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https://doi.org/10.1...
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
Ämnesord
Stäng
- Decoupling growth from product synthesis is a promising strategy to increase carbon partitioning and maximize productivity in cell factories. However, reduction in both substrate uptake rate and metabolic activity in the production phase are an underlying problem for upscaling. Here, we used CRISPR interference to repress growth in lactate-producing Synechocystis sp. PCC 6803. Carbon partitioning to lactate in the production phase exceeded 90%, but CO2 uptake was severely reduced compared to uptake during the growth phase. We characterized strains during the onset of growth arrest using transcriptomics and proteomics. Multiple genes involved in ATP homeostasis were regulated once growth was inhibited, which suggests an alteration of energy charge that may lead to reduced substrate uptake. In order to overcome the reduced metabolic activity and take advantage of increased carbon partitioning, we tested a novel production strategy that involved alternating growth arrest and recovery by periodic addition of an inducer molecule to activate CRISPRi. Using this strategy, we maintained lactate biosynthesis in Synechocystis for 30 days in a constant light turbidostat cultivation. Cumulative lactate titers were also increased by 100% compared to a constant growth-arrest regime, and reached 1 g/L. Further, the cultivation produced lactate for 30 days, compared to 20 days for the non-growth arrest cultivation. Periodic growth arrest could be applicable for other products, and in cyanobacteria, could be linked to internal circadian rhythms that persist in constant light.
Ämnesord
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
Nyckelord
- Two-stage production
- Cyanobacteria
- Stress response
- Synthetic biology
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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