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  In silico-guided engineering of Pseudomonas putida towards growth under micro-oxic conditions

Kampers, L. F. C., van Heck, R. G. A., Donati, S., Saccenti, E., Volkers, R. J. M., Schaap, P. J., Suarez-Diez, M., Nikel, P. I., & dos Santos, V. A. P. M. (2019). In silico-guided engineering of Pseudomonas putida towards growth under micro-oxic conditions. MICROBIAL CELL FACTORIES, 18(1):. doi:10.1186/s12934-019-1227-5.

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アイテムのパーマリンク: https://hdl.handle.net/21.11116/0000-0008-BEEE-7 版のパーマリンク: https://hdl.handle.net/21.11116/0000-0008-E3AE-4
資料種別: 学術論文

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 作成者:
Kampers, Linde F. C.1, 著者
van Heck, Ruben G. A.1, 著者
Donati, Stefano2, 著者           
Saccenti, Edoardo1, 著者
Volkers, Rita J. M.1, 著者
Schaap, Peter J.1, 著者
Suarez-Diez, Maria1, 著者
Nikel, Pablo I.1, 著者
dos Santos, Vitor A. P. Martins1, 著者
所属:
1external, ou_persistent22              
2Emmy Noether Research Group Dynamic Control of Metabolic Networks, Alumni, Max Planck Institute for Terrestrial Microbiology, Max Planck Society, Karl-von-Frisch-Strasse 10, D-35043 Marburg, DE, ou_3266292              

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 要旨: BackgroundPseudomonas putida is a metabolically versatile, genetically
accessible, and stress-robust species with outstanding potential to be
used as a workhorse for industrial applications. While industry
recognises the importance of robustness under micro-oxic conditions for
a stable production process, the obligate aerobic nature of P. putida,
attributed to its inability to produce sufficient ATP and maintain its
redox balance without molecular oxygen, severely limits its use for
biotechnology applications.ResultsHere, a combination of genome-scale
metabolic modelling and comparative genomics is used to pinpoint
essential O2-dependent processes. These explain the inability of the
strain to grow under anoxic conditions: a deficient ATP generation and
an inability to synthesize essential metabolites. Based on this, several
P. putida recombinant strains were constructed harbouring acetate kinase
from Escherichia coli for ATP production, and a class I dihydroorotate
dehydrogenase and a class III anaerobic ribonucleotide triphosphate
reductase from Lactobacillus lactis for the synthesis of essential
metabolites. Initial computational designs were fine-tuned by means of
adaptive laboratory evolution.ConclusionsWe demonstrated the value of
combining in silico approaches, experimental validation and adaptive
laboratory evolution for microbial design by making the strictly aerobic
Pseudomonas putida able to grow under micro-oxic conditions.

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 日付: 2019-10-22
 出版の状態: オンラインで出版済み
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 識別子(DOI, ISBNなど): ISI: 000491883800001
DOI: 10.1186/s12934-019-1227-5
 学位: -

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出版物 1

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出版物名: MICROBIAL CELL FACTORIES
種別: 学術雑誌
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出版社, 出版地: -
ページ: - 巻号: 18 (1) 通巻号: 179 開始・終了ページ: - 識別子(ISBN, ISSN, DOIなど): -