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 plas-mid 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 con-structed,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.