Construction of high-yield inositol-1-phosphate synthase recombinant Escherichia coli and optimization of fermentation conditions for myo-inositol production

In this study, the recombinant plasmid pETDuet-INO1 was constructed by introducing the expression gene INO1 of inositol-1-phosphate synthase (IPS) (a key enzyme in inositol synthesis pathway) cloned from Saccharomyces cerevisiae SC288 by gene engineering technology into plasmid pETDuet-1. The recombinant strain Escherichia coli BL21/pETDuet-INO1 was constructed by introducing the recombinant plasmid pETDuet- INO1 into E. coli BL21 to achieve efficient expression of IPS in E. coli. The fermentation conditions for myo-inositol production were optimized by single factor experiments and orthogonal experiments. The results showed that the recombinant E. coli BL21/pETDuet-INO1 was successfully constructed, and the optimal fermentation conditions for myo-inositol production were as follows: initial glucose addition 12 g/L, induction temperature 28 ℃, and inoculum 3%. The yield of myo-inositol was (0.81±0.007) g/L after induction 24 h under these conditions, which was 60.39% higher than that before optimization.

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Publication Details

Journal
SHILAP Revista de lepidopterología
Published
2024-12-01
DOI
https://doi.org/10.11882/j.issn.0254-5071.2024.12.021
Primary Topic
Mycorrhizal Fungi and Plant Interactions
Type
article
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article

Construction of high-yield inositol-1-phosphate synthase recombinant Escherichia coli and optimization of fermentation conditions for myo-inositol production

SHILAP Revista de lepidopterología
Mycorrhizal Fungi and Plant Interactions
article

Construction of high-yield inositol-1-phosphate synthase recombinant Escherichia coli and optimization of fermentation conditions for myo-inositol production

article en

Abstract

In this study, the recombinant plasmid pETDuet-INO1 was constructed by introducing the expression gene INO1 of inositol-1-phosphate synthase (IPS) (a key enzyme in inositol synthesis pathway) cloned from Saccharomyces cerevisiae SC288 by gene engineering technology into plasmid pETDuet-1. The recombinant strain Escherichia coli BL21/pETDuet-INO1 was constructed by introducing the recombinant plasmid pETDuet- INO1 into E. coli BL21 to achieve efficient expression of IPS in E. coli. The fermentation conditions for myo-inositol production were optimized by single factor experiments and orthogonal experiments. The results showed that the recombinant E. coli BL21/pETDuet-INO1 was successfully constructed, and the optimal fermentation conditions for myo-inositol production were as follows: initial glucose addition 12 g/L, induction temperature 28 ℃, and inoculum 3%. The yield of myo-inositol was (0.81±0.007) g/L after induction 24 h under these conditions, which was 60.39% higher than that before optimization.

SHILAP Revista de lepidopterología
Hangzhou Wanxiang Polytechnic (CN)
Openalex Percentile: Top 60%
Mycorrhizal Fungi and Plant Interactions
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