Biosynthesis of mannose from glucose via constructing phosphorylation-dephosphorylation reactions in Escherichia coli.

d-mannose has been widely used in food, medicine, cosmetic, and food-additive industries. To date, chemical synthesis or enzymatic conversion approaches based on iso/epimerization reactions for d-mannose production suffered from low conversion rate due to the reaction equilibrium, necessitating intricate separation processes for obtaining pure products on an industrial scale. To circumvent this challenge, this study showcased a new approach for d-mannose synthesis from glucose through constructing a phosphorylation-dephosphorylation pathway in an engineered strain. Specifically, the gene encoding phosphofructokinase (PfkA) in glycolytic pathway was deleted in Escherichia coli to accumulate fructose-6-phosphate (F6P). Additionally, one endogenous phosphatase, YniC, with high specificity to mannose-6-phosphate, was identified. In ΔpfkA strain, a recombinant synthetic pathway based on mannose-6-phosphate isomerase and YniC was developed to direct F6P to mannose. The resulting strain successfully produced 25.2 g/L mannose from glucose with a high conversion rate of 63% after transformation for 48 h. This performance surpassed the 15% conversion rate observed with 2-epimerases. In conclusion, this study presents an efficient method for achieving high-yield mannose synthesis from cost-effective glucose.

Vibration and Noise Analysis and Experimental Study of Rail Conveyor.

The rail conveyor is a new type of energy-saving system for the long-distance transportation of bulk materials. Operating noise is an urgent problem that the current model faces. It will cause noise pollution and affect the health of workers. In this paper, the factors causing vibration and noise are analyzed by modeling the wheel-rail system and the supporting truss structure. Based on the built test platform, the system vibration of the vertical steering wheel, the track support truss, and the track connection were measured, and the vibration characteristics at different positions were analyzed. Based on the established noise and vibration model, the distribution and occurrence rules of system noise under different operating speeds and fastener stiffness conditions were obtained. The experimental results show that the vibration amplitude of the frame near the head of the conveyor is the largest. The amplitude under the condition of 2 m/s running speed at the same position is 4 times that under the condition of 1 m/s. At different welds of the track, the width and depth of the rail gap have a great influence on the vibration impact, which is mainly due to the impact of the uneven impedance at the track gap, and the greater the running speed, the more obvious the vibration impact. The simulation results show the trend of noise generation, the speed of the trolley, and the stiffness of the track fasteners have a positive effect on the generation of noise in the low-frequency region. The research results of this paper will play an important role in the noise and vibration analysis of rail conveyors and help to optimize the structure design of the track transmission system.

Altered Cofactor Preference of Thermostable StDAPDH by a Single Mutation at K159.

D-amino acid production from 2-keto acid by reductive amination is an attractive pathway because of its high yield and environmental safety. StDAPDH, a meso-diaminopimelate dehydrogenase (meso-DAPDH) from Symbiobacterium thermophilum, was the first meso-DAPDH to show amination of 2-keto acids. Furthermore, StDAPDH shows excellent thermostability compared to other meso-DAPDHs. However, the cofactor of StDAPDH is NADP(H), which is less common than NAD(H) in industrial applications. Therefore, cofactor engineering for StDAPDH is needed. In this study, the highly conserved cofactor binding sites around the adenosine moiety of NADPH were targeted to determine cofactor specificity. Lysine residues within a loop were found to be critical for the cofactor specificity of StDAPDH. Replacement of lysine with arginine resulted in the activity of pyruvic acid with NADH as the cofactor. The affinity of K159R to pyruvic acid was equal with NADH or NADPH as the cofactor, regardless of the mutation. Molecular dynamics simulations revealed that the large steric hindrance of arginine and the interaction of the salt bridge between NADH and arginine may have restricted the free movement of NADH, which prompted the formation of a stable active conformation of mutant K159R. These results provide further understanding of the catalytic mechanism of StDAPDH and guidance for the cofactor engineering of StDAPDH.

Cinnabar is not converted into methylmercury by human intestinal bacteria.

ETHNOPHARMACOLOGICAL RELEVANCE: Cinnabar (Cin), a naturally occurring mercuric sulfide (HgS), is a mineral widely used in traditional Chinese medicine throughout history. As for the toxicity of cinnabar, one important assumption is that cinnabar may be transformed into highly toxic methylmercury by gastrointestinal flora. There is no evidence in humans to support this assumption. AIM OF THE STUDY: To investigate the biotransformation of cinnabar (HgS) in the human intestinal bacteria with modern analytical techniques. MATERIALS AND METHODS: A gas chromatograph, equipped with electron capture detection (GC-ECD) and mass spectrometry (GC-MS), were used to detect the formation of methylmercury after incubation of cinnabar with human intestinal bacteria. The content of soluble mercury in the bacteria media was determined by cold vapor-atomic absorption spectrometry (CV-AAS). In addition, X-ray absorption near-edge structure spectroscopy (XANES) was used to confirm the possible transformation of cinnabar in the bacteria media, and under mimetic intestinal condition by measuring the species of sulfur and mercury in the reaction extraction of cinnabar and Na(2)S mixture. RESULTS: No methylmercury was detected by both GC-ECD and GC-MS, which suggest that cinnabar (HgS) is not methylated in the human intestine. A small amount of soluble mercury was found to be released in the flora medium of HgS or cinnabar by CV-AAS. The XANES analyses revealed that polysulfides exist in the flora medium, and the simulated results showed that the products by incubating cinnabar with Na(2)S were mercuric polysulfides. CONCLUSION: These results showed that under gut flora conditions cinnabar would be transformed into mercuric polysulfides rather than methylmercury. Our work provides evidences of nontoxic transformation of cinnabar in the human intestinal bacteria.

Efficiency of methods applied for goat estrous synchronization in subtropical monsoonal climate zone of Southwest China.

Two experiments were undertaken to select the efficient method applied for goat estrous synchronization. In experiment 1, a total of 120 does (Capra hircus) were divided into five groups with a randomized block design, and the does of treatment 1 were synchronized by intravaginal sponges impregnated with 30 mg Levonorgestrel inserted for 10 days. Does of treatments 2, 3, and 4 were treated with further injection of 25 IU follicle-stimulating hormone (FSH), 0.05 mg prostaglandin F2 alpha (PGF(2alpha)), and 25 IU FSH + 0.05 mg PGF(2alpha) at sponge withdrawal, respectively. The does in the control group (n = 40) without estrous synchronization treatment and natural estrous does were observed. In experiment 2, a total of 140 does in five goat farms in breeding and non-breeding seasons were treated with the selective efficient procedure. The results presented that all the employed treatments were capable of inducing and synchronizing estrous goats. According to estrous response and economy, the use of intravaginal sponges impregnated with 30 mg Levonorgestrel and 0.05 mg PGF(2alpha) (treatment 3) is the first choice for estrous synchronization, and 95.0% of synchronized does demonstrated estrus, which was significantly higher than that of treatment 1 (P < 0.05) and control group (P < 0.01). The percentages of ovulating of treatments 3 and 4 were the same (95.0%), which were significantly higher than that of treatment 1 (P < 0.01). The ovulation rates among different groups were not significant (P > 0.05). When the selective procedure was applied to five goat farms, 85.7% (120/140) of does demonstrated estrus, and the kidding percentage, litter size, and prolificacy rate were 53.6%, 0.95, and 177%, respectively.