Ding, Xiumin published the artcileBottom-up synthetic biology approach for improving the efficiency of menaquinone-7 synthesis in Bacillus subtilis, Product Details of C5H11NO2S, the publication is Microbial Cell Factories (2022), 21(1), 101, database is CAplus and MEDLINE.
Menaquinone-7 (MK-7), which is associated with complex and tightly regulated pathways and redox imbalances, is produced at low titers in Bacillus subtilis. Synthetic biol. provides a rational engineering principle for the transcriptional optimization of key enzymes and the artificial creation of cofactor regeneration systems without regulatory interference. This holds great promise for alleviating pathway bottlenecks and improving the efficiency of carbon and energy utilization. We used a bottom-up synthetic biol. approach for the synthetic redesign of central carbon and to improve the adaptability between material and energy metabolism in MK-7 synthesis pathways. First, the rate-limiting enzymes, 1-deoxyxylulose-5-phosphate synthase (DXS), isopentenyl-diphosphate delta-isomerase (Fni), 1-deoxyxylulose-5-phosphate reductase (DXR), isochorismate synthase (MenF), and 3-deoxy-7-phosphoheptulonate synthase (AroA) in the MK-7 pathway were sequentially overexpressed. Promoter engineering and fusion tags were used to overexpress the key enzyme MenA, and the titer of MK-7 was 39.01 mg/L. Finally, after stoichiometric calculation and optimization of the cofactor regeneration pathway, we constructed two NADPH regeneration systems, enhanced the endogenous cofactor regeneration pathway, and introduced a heterologous NADH kinase (Pos5P) to increase the availability of NADPH for MK-7 biosynthesis. The strain expressing pos5P was more efficient in converting NADH to NADPH and had excellent MK-7 synthesis ability. Following three Design-Build-Test-Learn cycles, the titer of MK-7 after flask fermentation reached 53.07 mg/L, which was 4.52 times that of B. subtilis 168. Addnl., the artificially constructed cofactor regeneration system reduced the amount of NADH-dependent byproduct lactate in the fermentation broth by 9.15%. This resulted in decreased energy loss and improved carbon conversion. In summary, a “high-efficiency, low-carbon, cofactor-recycling” MK-7 synthetic strain was constructed, and the strategy used in this study can be generally applied for constructing high-efficiency synthesis platforms for other terpenoids, laying the foundation for the large-scale production of high-value MK-7 as well as terpenoids.
Microbial Cell Factories published new progress about 63-68-3. 63-68-3 belongs to catalysis-chemistry, auxiliary class Natural product, name is (S)-2-Amino-4-(methylthio)butanoic acid, and the molecular formula is C5H11NO2S, Product Details of C5H11NO2S.
Referemce:
https://courses.lumenlearning.com/boundless-chemistry/chapter/catalysis/,
Catalysis – Wikipedia