MOESM4 of Production of trehalose with trehalose synthase expressed and displayed on the surface of Bacillus subtilis spores

Additional file 4. Construction of recombinant plasmids pDG1730-CotC-treS and pDG1730-CotG-treS

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Additional file 6. The strategy for the chromosomal integration of the Cot-treS fusion genes

MOESM1 of Production of trehalose with trehalose synthase expressed and displayed on the surface of Bacillus subtilis spores

Additional file 1. The dot blot assay of TreS displayed on spore surface from different recombinant bacteria. 1: TreS standard; C+G: TreS displayed on the spore surface of B. subtilis WB800n/cotC-treS–cotG-treS; C: TreS displayed on the spore surface of B. subtilis WB800n/cotC-treS; G: TreS displayed on the spore surface of B. subtilis WB800n/cotG-treS

MOESM2 of Production of trehalose with trehalose synthase expressed and displayed on the surface of Bacillus subtilis spores

Additional file 2. Optimal pH, temperature, and tolerance of TreS displayed on the spore surface

MOESM7 of Production of trehalose with trehalose synthase expressed and displayed on the surface of Bacillus subtilis spores

Additional file 7. Construction of recombinant B. subtilis WB800n with deletion of genes sleB and cwlJ

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Additional file 5. Construction of recombinant plasmid pDG1730-CotC-treS–CotG-treS

MOESM3 of Production of trehalose with trehalose synthase expressed and displayed on the surface of Bacillus subtilis spores

Additional file 3. The bacterial strains, plasmids, and primers used in this study

An inventory rotation mechanism for relief supplies considering recycling and remanufacturing

Relief supply inventories are important in disaster response operations. To address the problem of dead stocks and avoid relief supply expiration, such inventories must be renewed periodically. Different from the traditional shelf-life rotation mechanism (i.e. renewal of the entire inventory at the expiration date), which pays more attention to decreasing the quantity of dead stocks, the novel period rotation mechanism that we design in this study will gradually rotate relief supplies according to their shelf life. We propose a mathematical programming model to minimise the total cost of relief organisations and conduct a case study and analyse the sensitivity of key parameters using real-world data. Results show that our rotation mechanism can substantially reduce the financial burden on relief organisations and improve the quality of relief supplies for victims. The novelty of our study includes: (1) We introduce a new approach that can rotate multiple times, through which we consider the quality of relief supplies; (2) We incorporate recycling and remanufacturing into the rotation process; and (3) We adopt an inductive reasoning method to develop the model with demand and time uncertainties. We prove the feasibility of our model and contributions of our study to humanitarian logistics practice and research.</p

Balancing Pyruvate Node Based on a Dual-Layered Dynamic Regulation System to Improve the Biosynthesis of Caffeic Acid in Candida glycerinogenes

Caffeic acid is a phenolic acid compound widely applied in the food and pharmaceutical fields. Currently, one of the reasons for the low yield of caffeic acid biosynthesis is that the carbon flow enters mainly into the TCA cycle via pyruvate, which leads to low concentrations of erythrose 4-phosphate (E4P) and phosphoenolpyruvate (PEP), the precursors of caffeic acid synthesis. Here, we developed a growth-coupled dual-layered dynamic regulation system. This system controls intracellular pyruvate supply in real time by responding to intracellular pyruvate and p-coumaric acid concentrations, autonomously coordinates pathway gene expression, and redirects carbon metabolism to balance cell growth and caffeic acid synthesis. Finally, our constructed engineered strain based on the dual-layered dynamic regulation system achieved a caffeic acid titer of 559.7 mg/L in a 5 L bioreactor. Thus, this study demonstrated the efficiency and potential of this system in boosting the yield of aromatic compounds

Data_Sheet_1_Development of a co-culture system for green production of caffeic acid from sugarcane bagasse hydrolysate.PDF

Caffeic acid (CA) is a phenolic acid compound widely used in pharmaceutical and food applications. However, the efficient synthesis of CA is usually limited by the resources of individual microbial platforms. Here, a cross-kingdom microbial consortium was developed to synthesize CA from sugarcane bagasse hydrolysate using Escherichia coli and Candida glycerinogenes as chassis. In the upstream E. coli module, shikimate accumulation was improved by intensifying the shikimate synthesis pathway and blocking shikimate metabolism to provide precursors for the downstream CA synthesis module. In the downstream C. glycerinogenes module, conversion of p-coumaric acid to CA was improved by increasing the supply of the cytoplasmic cofactor FAD(H2). Further, overexpression of ABC transporter-related genes promoted efflux of CA and enhanced strain resistance to CA, significantly increasing CA titer from 103.8 mg/L to 346.5 mg/L. Subsequently, optimization of the inoculation ratio of strains SA-Ec4 and CA-Cg27 in this cross-kingdom microbial consortium resulted in an increase in CA titer to 871.9 mg/L, which was 151.6% higher compared to the monoculture strain CA-Cg27. Ultimately, 2311.6 and 1943.2 mg/L of CA were obtained by optimization of the co-culture system in a 5 L bioreactor using mixed sugar and sugarcane bagasse hydrolysate, respectively, with 17.2-fold and 14.6-fold enhancement compared to the starting strain. The cross-kingdom microbial consortium developed in this study provides a reference for the production of other aromatic compounds from inexpensive raw materials.</p