التفاصيل البيبلوغرافية
| العنوان: |
Model-guided metabolic rewiring to bypass pyruvate oxidation for pyruvate derivative synthesis by minimizing carbon loss |
| المؤلفون: |
Zhang, Yun, Wang, Xueliang, Odesanmi, Christianah, Hu, Qitiao, Li, Dandan, Tang, Yuan, Liu, Zhe, Mi, Jie, Liu, Shuwen, Wen, Tingyi |
| المساهمون: |
Gilbert, Jack A., MOST | National Natural Science Foundation of China, MOST | National Key Research and Development Program of China |
| المصدر: |
mSystems ; volume 9, issue 3 ; ISSN 2379-5077 |
| بيانات النشر: |
American Society for Microbiology |
| سنة النشر: |
2024 |
| الوصف: |
Engineering microbial hosts to synthesize pyruvate derivatives depends on blocking pyruvate oxidation, thereby causing severe growth defects in aerobic glucose-based bioprocesses. To decouple pyruvate metabolism from cell growth to improve pyruvate availability, a genome-scale metabolic model combined with constraint-based flux balance analysis, geometric flux balance analysis, and flux variable analysis was used to identify genetic targets for strain design. Using translation elements from a ~3,000 cistronic library to modulate fxpK expression in a bicistronic cassette, a bifido shunt pathway was introduced to generate three molecules of non-pyruvate-derived acetyl-CoA from one molecule of glucose, bypassing pyruvate oxidation and carbon dioxide generation. The dynamic control of flux distribution by T7 RNAP-mediated synthetic small RNA decoupled pyruvate catabolism from cell growth. Adaptive laboratory evolution and multi-omics analysis revealed that a mutated isocitrate dehydrogenase functioned as a metabolic switch to activate the glyoxylate shunt as the only C4 anaplerotic pathway to generate malate from two molecules of acetyl-CoA input and bypass two decarboxylation reactions in the tricarboxylic acid cycle. A chassis strain for pyruvate derivative synthesis was constructed to reduce carbon loss by using the glyoxylate shunt as the only C4 anaplerotic pathway and the bifido shunt as a non-pyruvate-derived acetyl-CoA synthetic pathway and produced 22.46, 27.62, and 6.28 g/L of l -leucine, l -alanine, and l -valine by a controlled small RNA switch, respectively. Our study establishes a novel metabolic pattern of glucose-grown bacteria to minimize carbon loss under aerobic conditions and provides valuable insights into cell design for manufacturing pyruvate-derived products. IMPORTANCE Bio-manufacturing from biomass-derived carbon sources using microbes as a cell factory provides an eco-friendly alternative to petrochemical-based processes. Pyruvate serves as a crucial building block for the ... |
| نوع الوثيقة: |
article in journal/newspaper |
| اللغة: |
English |
| DOI: |
10.1128/msystems.00839-23 |
| الإتاحة: |
https://doi.org/10.1128/msystems.00839-23Test |
| حقوق: |
https://creativecommons.org/licenses/by/4.0Test/ ; https://journals.asm.org/non-commercial-tdm-licenseTest |
| رقم الانضمام: |
edsbas.8C72C400 |
| قاعدة البيانات: |
BASE |
