論文 - 詳細
| RRC ID | 27479 |
|---|---|
| 著者 | Hasegawa S, Suda M, Uematsu K, Natsuma Y, Hiraga K, Jojima T, Inui M, Yukawa H. |
| タイトル | Engineering of Corynebacterium glutamicum for high-yield L-valine production under oxygen deprivation conditions. |
| ジャーナル | Appl Environ Microbiol |
| Abstract |
We previously demonstrated efficient L-valine production by metabolically engineered Corynebacterium glutamicum under oxygen deprivation. To achieve the high productivity, a NADH/NADPH cofactor imbalance during the synthesis of l-valine was overcome by engineering NAD-preferring mutant acetohydroxy acid isomeroreductase (AHAIR) and using NAD-specific leucine dehydrogenase from Lysinibacillus sphaericus. Lactate as a by-product was largely eliminated by disrupting the lactate dehydrogenase gene ldhA. Nonetheless, a few other by-products, particularly succinate, were still produced and acted to suppress the L-valine yield. Eliminating these by-products therefore was deemed key to improving theL-valine yield. By additionally disrupting the phosphoenolpyruvate carboxylase gene ppc, succinate production was effectively suppressed, but both glucose consumption and L-valine production dropped considerably due to the severely elevated intracellular NADH/NAD(+) ratio. In contrast, this perturbed intracellular redox state was more than compensated for by deletion of three genes associated with NADH-producing acetate synthesis and overexpression of five glycolytic genes, including gapA, encoding NADH-inhibited glyceraldehyde-3-phosphate dehydrogenase. Inserting feedback-resistant mutant acetohydroxy acid synthase and NAD-preferring mutant AHAIR in the chromosome resulted in higher L-valine yield and productivity. Deleting the alanine transaminase gene avtA suppressed alanine production. The resultant strain produced 1,280 mM L-valine at a yield of 88% mol mol of glucose(-1) after 24 h under oxygen deprivation, a vastly improved yield over our previous best. |
| 巻・号 | 79(4) |
| ページ | 1250-7 |
| 公開日 | 2013-2-1 |
| DOI | 10.1128/AEM.02806-12 |
| PII | AEM.02806-12 |
| PMID | 23241971 |
| PMC | PMC3568611 |
| MeSH | Anaerobiosis Biosynthetic Pathways / genetics* Corynebacterium glutamicum / genetics* Corynebacterium glutamicum / metabolism* Gene Deletion Gene Expression Lactic Acid / metabolism Metabolic Engineering* NAD / metabolism Oxygen / metabolism Recombination, Genetic Succinic Acid / metabolism Valine / biosynthesis* |
| IF | 4.016 |
| 引用数 | 71 |
| WOS 分野 | BIOTECHNOLOGY & APPLIED MICROBIOLOGY MICROBIOLOGY |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | Patent(IFI CLAIMS) |
| 各媒体での言及数の合計 | 16 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| 一般微生物 | JCM 18229 |