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| LEADER |
02901nam a2200505Ia 4500 |
| 001 |
10.1186-s13068-021-01954-6 |
| 008 |
220427s2021 CNT 000 0 und d |
| 020 |
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|a 17546834 (ISSN)
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| 245 |
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|a ATP and NADPH engineering of Escherichia coli to improve the production of 4-hydroxyphenylacetic acid using CRISPRi
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| 260 |
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|b BioMed Central Ltd
|c 2021
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| 856 |
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|z View Fulltext in Publisher
|u https://doi.org/10.1186/s13068-021-01954-6
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|a Background: 4-Hydroxyphenylacetic acid (4HPAA) is an important raw material for the synthesis of drugs, pesticides and biochemicals. Microbial biotechnology would be an attractive approach for 4HPAA production, and cofactors play an important role in biosynthesis. Results: We developed a novel strategy called cofactor engineering based on clustered regularly interspaced short palindromic repeat interference (CRISPRi) screening (CECRiS) for improving NADPH and/or ATP availability, enhancing the production of 4HPAA. All NADPH-consuming and ATP-consuming enzyme-encoding genes of E. coli were repressed through CRISPRi. After CRISPRi screening, 6 NADPH-consuming and 19 ATP-consuming enzyme-encoding genes were identified. The deletion of the NADPH-consuming enzyme-encoding gene yahK and the ATP-consuming enzyme-encoding gene fecE increased the production of 4HPAA from 6.32 to 7.76 g/L. Automatically downregulating the expression of the pabA gene using the Esa-PesaS quorum-sensing-repressing system further improved the production of 4HPAA. The final strain E. coli 4HPAA-∆yfp produced 28.57 g/L of 4HPAA with a yield of 27.64% (mol/mol) in 2-L bioreactor fed-batch fermentations. The titer and yield are the highest values to date. Conclusion: This CECRiS strategy will be useful in engineering microorganisms for the high-level production of bioproducts. © 2021, The Author(s).
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|a 4-Hydroxyphenylacetic acid
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|a ATP engineering
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|a Biochemistry
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|a Bioproducts
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|a biotechnology
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|a carboxylic acid
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|a Cofactor engineering
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|a coliform bacterium
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|a CRISPRi
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|a Encoding (symbols)
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|a Encoding genes
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|a enzyme
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|a Enzymes
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|a Escherichia coli
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|a Escherichia coli
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|a Escherichia coli
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|a Fed-batch fermentation
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|a Genes
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| 650 |
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|a genetic engineering
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|a High-level production
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|a Microbial biotechnology
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|a NADPH engineering
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| 650 |
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|a Novel strategies
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| 650 |
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|a Palindromic
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| 650 |
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|a Quorum-sensing
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|a Signal encoding
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| 700 |
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|a He, X.
|e author
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|a Liao, Y.-L.
|e author
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|a Liu, J.-Z.
|e author
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|a Lu, Q.
|e author
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|a Shen, Y.-P.
|e author
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| 700 |
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|a Yan, Z.-B.
|e author
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| 773 |
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|t Biotechnology for Biofuels
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