| Products/Services Used | Details | Operation |
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| Gene Synthesis | Samples in the exponential phase were collected and immediately centrifuged at 10,000 × g and 4 °C for 5 min. Total RNA was isolated using the TaKaRa RNAiso Plus (Takara). The RNA was reverse transcribed using a cDNA synthesis kit (Takara) according to the manufacturer’s protocol. Primers were designed using an online tool (https://www.genscript.com/ssl-bin/app/primer). Appropriately diluted cDNA was used with a SYBRs Premix Ex TaqTM kit (Takara) according to the manufacturer’s protocol. The expression levels of the selected genes were determined using the ABI 7500 Real-Time PCR system (Applied Biosystems, USA) with the primers listed in Table 2. The 16S rRNA gene was used as the housekeeping gene (internal control). The relative fold change in gene expression was calculated by the comparative Ct method (2−ΔΔCt method) [26]. All real-time PCRs were performed as three independent experiments. | Get A Quote |
Bio-based succinate synthesis process suffers from low production efficiency, especially in the late stage of fermentation because of high osmotic stress. In this study, DR1558 (a response regulator in tow-component signal transduction systems) from Deinococcus radiodurans was overexpressed in Escherichia coli Suc260. High concentration glucose and NaCl impact experiment results showed that the recombinant strain (E. coli Suc270) displayed higher tolerance to osmotic stress. Fermentation in a 3-L fermenter showed that the dry cell weight, glucose consumption, and succinic acid titer of Suc270 were 2.06, 90.2, and 55.4 g/L, which is an increase of 1.50-, 1.38-, and 1.26-fold, respectively, compared to thos... More
