Enhancing the Ability of Escherichia coli to Synthesise L-Isoleucine Using λ-Red Recombinant Technology Combined with Complex Mutagenesis
To improve the synthetic ability of E. coli L-isoleucine by combining λ-Red recombination with complex mutagenesis. Firstly, taking E. coli NXA as the original strain, λ-Red homologous recombination was used to knock out the coding gene brnQ of branched chain amino acid transport protein to obtain m...
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| Formato: | Artigo |
| Idioma: | zho |
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The editorial department of Science and Technology of Food Industry
2025-01-01
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| Colecção: | Shipin gongye ke-ji |
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| Acesso em linha: | http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2024020234 |
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| author | Jipeng WANG Tengteng ZHU Lu LIU Cheng MA Xiaobo WEI Huiyan LIU Haitian FANG |
| author_facet | Jipeng WANG Tengteng ZHU Lu LIU Cheng MA Xiaobo WEI Huiyan LIU Haitian FANG |
| author_sort | Jipeng WANG |
| collection | DOAJ |
| description | To improve the synthetic ability of E. coli L-isoleucine by combining λ-Red recombination with complex mutagenesis. Firstly, taking E. coli NXA as the original strain, λ-Red homologous recombination was used to knock out the coding gene brnQ of branched chain amino acid transport protein to obtain mutant strain E. coli NXA1. Secondly, E. coli NXA1 was subjected to multiple rounds of complex mutagenesis with atmospheric room temperature plasma (ARTP), ultraviolet (UV), and nitrosoguanidine (NTG), which was screened to obtain the mutant strain E. coli NXA2 with α-AB being structural analogue. The fermentation results showed that L-isoleucine titer of E. coli NXA1 was 2.76 g/L, which was 33.98% higher than E. coli NXA after fermentation for 40 hours at 37 ℃ and 200 r/min. The L-isoleucine titer of E. coli NXA2 was 3.22 g/L, which was 16.67% higher than E. coli NXA1 and 56.31% higher than E. coli NXA. After 20 continuous passages of E. coli NXA2, the good genetic stability could be reflected. The combination of λ-Red recombination and complex mutagenesis has a significant effect on improving the synthetic ability of E. coli L-isoleucine, laying a theoretical basis for the breeding of L-isoleucine high-producing strains. |
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| institution | Directory Open Access Journal |
| issn | 1002-0306 |
| language | zho |
| last_indexed | 2025-02-17T03:53:22Z |
| publishDate | 2025-01-01 |
| publisher | The editorial department of Science and Technology of Food Industry |
| record_format | Article |
| series | Shipin gongye ke-ji |
| spelling | doaj.art-0da78134e893435287f901b4fb9453d22025-01-10T06:49:30ZzhoThe editorial department of Science and Technology of Food IndustryShipin gongye ke-ji1002-03062025-01-0146216717410.13386/j.issn1002-0306.20240202342024020234-2Enhancing the Ability of Escherichia coli to Synthesise L-Isoleucine Using λ-Red Recombinant Technology Combined with Complex MutagenesisJipeng WANG0Tengteng ZHU1Lu LIU2Cheng MA3Xiaobo WEI4Huiyan LIU5Haitian FANG6Ningxia Key Laboratory of Food Microbial Application Technology and Safety Control, School of Food Science and Engineering, Ningxia University, Yinchuan 750021, ChinaNingxia Key Laboratory of Food Microbial Application Technology and Safety Control, School of Food Science and Engineering, Ningxia University, Yinchuan 750021, ChinaNingxia Key Laboratory of Food Microbial Application Technology and Safety Control, School of Food Science and Engineering, Ningxia University, Yinchuan 750021, ChinaNingxia 36du Biotechnology Co., Ltd., Yinchuan 750021, ChinaNingxia Key Laboratory of Food Microbial Application Technology and Safety Control, School of Food Science and Engineering, Ningxia University, Yinchuan 750021, ChinaNingxia Key Laboratory of Food Microbial Application Technology and Safety Control, School of Food Science and Engineering, Ningxia University, Yinchuan 750021, ChinaNingxia Key Laboratory of Food Microbial Application Technology and Safety Control, School of Food Science and Engineering, Ningxia University, Yinchuan 750021, ChinaTo improve the synthetic ability of E. coli L-isoleucine by combining λ-Red recombination with complex mutagenesis. Firstly, taking E. coli NXA as the original strain, λ-Red homologous recombination was used to knock out the coding gene brnQ of branched chain amino acid transport protein to obtain mutant strain E. coli NXA1. Secondly, E. coli NXA1 was subjected to multiple rounds of complex mutagenesis with atmospheric room temperature plasma (ARTP), ultraviolet (UV), and nitrosoguanidine (NTG), which was screened to obtain the mutant strain E. coli NXA2 with α-AB being structural analogue. The fermentation results showed that L-isoleucine titer of E. coli NXA1 was 2.76 g/L, which was 33.98% higher than E. coli NXA after fermentation for 40 hours at 37 ℃ and 200 r/min. The L-isoleucine titer of E. coli NXA2 was 3.22 g/L, which was 16.67% higher than E. coli NXA1 and 56.31% higher than E. coli NXA. After 20 continuous passages of E. coli NXA2, the good genetic stability could be reflected. The combination of λ-Red recombination and complex mutagenesis has a significant effect on improving the synthetic ability of E. coli L-isoleucine, laying a theoretical basis for the breeding of L-isoleucine high-producing strains.http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2024020234escherichia coliλ-red recombinant technologybrnq genecomplex mutagenesisfermentationl-isoleucine |
| spellingShingle | Jipeng WANG Tengteng ZHU Lu LIU Cheng MA Xiaobo WEI Huiyan LIU Haitian FANG Enhancing the Ability of Escherichia coli to Synthesise L-Isoleucine Using λ-Red Recombinant Technology Combined with Complex Mutagenesis Shipin gongye ke-ji escherichia coli λ-red recombinant technology brnq gene complex mutagenesis fermentation l-isoleucine |
| title | Enhancing the Ability of Escherichia coli to Synthesise L-Isoleucine Using λ-Red Recombinant Technology Combined with Complex Mutagenesis |
| title_full | Enhancing the Ability of Escherichia coli to Synthesise L-Isoleucine Using λ-Red Recombinant Technology Combined with Complex Mutagenesis |
| title_fullStr | Enhancing the Ability of Escherichia coli to Synthesise L-Isoleucine Using λ-Red Recombinant Technology Combined with Complex Mutagenesis |
| title_full_unstemmed | Enhancing the Ability of Escherichia coli to Synthesise L-Isoleucine Using λ-Red Recombinant Technology Combined with Complex Mutagenesis |
| title_short | Enhancing the Ability of Escherichia coli to Synthesise L-Isoleucine Using λ-Red Recombinant Technology Combined with Complex Mutagenesis |
| title_sort | enhancing the ability of escherichia coli to synthesise l isoleucine using λ red recombinant technology combined with complex mutagenesis |
| topic | escherichia coli λ-red recombinant technology brnq gene complex mutagenesis fermentation l-isoleucine |
| url | http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2024020234 |
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