In this research, a recombinant Escherichia coli strain BL21(TPP) was constructed to synthesize malonate through overexpressing six genes of ppc, aspC, panD, pa0132, yneI and pyc. Under shake flask fermentation conditons, strain BL21(TPP) produced 0.61 g/L malonic acid. In a 5 L fermentor, the production of malonic acid reached 3.32 g/L making use of an intermittent feeding method. Then, a recombinant strain BL21(SCR) had been constructed by fusional phrase of ppc and aspC, aswell as pa0132 and yneI, respectively. As a result, the production of malonic acid increased to 0.83 g/L in the shake flask level, that has been a 36% boost on the beginning strain BL21(TPP). Finally, the best malonate production reached 5.61 g/L in a 5 L fermentor, that was a 69% boost throughout the starting strain BL21(TPP). Creation of malonic acid by metabolically designed E. coli provides a basis for further optimization, and may serve as a reference when it comes to biosynthesis of various other dicarboxylic acids.D-mannitol is widely used into the pharmaceutical and medical sectors as an important precursor of antitumor medicines and resistant stimulants. However, the expense of the present enzymatic process for D-mannitol synthesis is high, thus perhaps not suited to commercialization. To handle this problem, an efficient mannitol dehydrogenase LpGDH used for the transformation and a glucose dehydrogenase BaGDH employed for NADH regeneration were screened, respectively. Those two enzymes had been LY333531 co-expressed in Escherichia coli BL21(DE3) to create a two-enzyme cascade catalytic response for the efficient synthesis of d-mannitol, with a conversion rate of 59.7% from D-fructose achieved. The regeneration of cofactor NADH had been improved by enhancing the copy wide range of Bagdh, and a recombinant strain E. coli BL21/pETDuet-Lpmdh-Bagdh-Bagdh was constructed to deal with the imbalance between cofactor quantity and key enzyme expression degree within the two-enzyme cascade catalytic reaction. An optimized whole mobile change procedure had been carried out under 30 ℃, initial pH 6.5, cell size (OD600) 30, 100 g/L D-fructose substrate and an equivalent molar concentration of sugar. The greatest yield of D-mannitol was 81.9 g/L with a molar conversion rate of 81.9% in 5 L fermenter under the ideal transformation problems. This research provides a green and efficient biotransformation way of future large-scale creation of D-mannitol, which is also of good relevance for the production of other sugar alcohols.Natamycin is a natural, broad spectrum and extremely efficient antifungal substance that belongs to polyene macrolide antibiotics. It was found in avoidance of food fungal contamination and treatment of clinical fungal infection. The extracellular transportation performance of natamycin are an important facet hampering the yield of natamycin produced by Streptomyces gilvosporeus. The extracellular transporter SgnA/B of natamycin ended up being analyzed by bioinformatics tools and molecular docking practices. This ATP-binding cassette transporter, contained SgnA and SgnB, is a heterodimers with inward-facing conformation. The essential difference between the natamycin combining efficiency of the two drug-binding cavities in SgnA/B is positive for natamycin extracellular transport. sgnA/B gene had been overexpressed in S. gilvosporeus F607 and the effects of sgnA/B gene overexpression on natamycin synthesis and extracellular transportation were reviewed. In F-EX strain, the extracellular/intracellular ratio of natamycin in logarithmic synthesis phase was increased, and the total fermentation yield at 120 h had been increased by 12.5per cent and reached to 7.38 g/L. Moreover, transcriptome sequencing analysis indicated that sgnA/B gene overexpression impacted the expression of genetics mixed up in k-calorie burning of numerous amino acids, propionate, glucose, C5-branched dibasic acid and TCA cycle. This research demonstrated that the improved extracellular transportation increased the synthesis of natamycin by S. gilvosporeus, and S. gilvosporeus F-EX showed great prospect of the industrial production of natamycin.L-ascorbic acid 2-glucoside (AA-2G) is a derivative of L-ascorbic acid (L-AA). In contrast to L-AA, it’s great stability and is Biotic indices easily decomposed by chemical in the human body. α-Glucosidase (AG) had been initial chemical found effective at producing AA-2G. But, researches with this chemical continues to be in infancy. We took AG produced from Aspergillus niger (AAG), Japanese rice (JrAG) and Rattus rattus (RAG), and compared their particular specific enzymatic task and transglycosidation rate, because of the seek to improve synthesis of AA-2G because of the transglycosidation of AG. The genetics encoding these three various AG had been cloned and expressed in engineered fungus. The circumstances for the transglycosidation result of these three enzymes were enhanced and also the transglycosidation performance and yield of AA-2G underneath the optimized circumstances had been compared. The specific activity of AAG achieved 1.0 U/mg, although the yield of AA-2G reached 153.1 mg/L with a transglycosidation rate of 0.5per cent. The precise activity of RAG achieved 0.4 U/mg, while the yield of AA-2G achieved 861.0 mg/L with a transglycosidation price of 2.5%. JrAG showed the greatest particular activity and transglycosidation rate. The enzyme specific activity of JrAG reached 1.9 U/mg, while the yield of AA-2G achieved 2 577.2 mg/L with a transglycosidation price of 7.6per cent, a lot higher than compared to the other two glucosidases. JrAG may hence have possible to boost the synthesis of AA-2G.A bio-electrochemical system can promote the interaction between microorganism and electrode and consequently alter cellular metabolic process. To research the metabolic performance of Zymomonas mobilis into the bio-electrochemical system, we applied an H-type bio-electrochemical reactor to control Z. mobilis fermentation under 3 V. Compared with the control team without used current teaching of forensic medicine , the glycerol within the anode chamber increased by 24%, although the glucose consumption into the cathode chamber increased by 16%, and the ethanol and succinic acid concentration increased by 13% and 8%, correspondingly.