ABSTRACT As one of the essential amino acids,厶-valine plays an important role in physioloxy and biochemistro. Pyruvate io not only the precursor needed for 厶-valine biosynthesis, but also an impoSani intermediate metabolite. Therefore, the efficient accumulation of 厶-valine and cell growth are mutually affected. In order te keep balancc between cell growth and accumulation of 厶-valine, the CRISPR interferencc technique Oo Coybaceium glutamate was established, and the relativa transcription leaeis of sevarai genes in tricarboxylio acid cycle % TCA) and pentoss phosphate pathway % PPP) were reeulated in varying deerees. Through shaking flask experiment, the genes aceE enccding pyruvate dehydroxenase and Cg/1576 enccding glucase-6-phosphate dehydroxenation were confirmed as eXXctive modiVcation taroetr fo 厶-valine accumulation. When the initiation of aceE and Cg/1576 were weekened, 34.6 and 32.3 g/L 厶-valine were accumulated, which were 6. 1 and 3. 8 g/L higher than the controi, respectivaly. In addition, O was conOrmed that weakening the initiation of aceE and Cg/1576 was the best interfeI■enco strateey, with a final !-valine accumulation of 37. 1 g/L. Thio study not only determined important taroet genes for modification, but also provided a new direction foe accumulation of 厶-valine in C. glutamate.

Key words Corynebacterium glutamicum ；厶-valine ； CRISPRi

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