Metabolic engineering of Escherichia coli for efficient production of L-5-hydroxytryptophan from glucose.

BACKGROUND: 5-hydroxytryptophan (5-HTP), the direct biosynthetic precursor of the neurotransmitter 5-hydroxytryptamine, has been shown to have unique efficacy in the treatment of a variety of disorders, including depression, insomnia, and chronic headaches, and is one of the most commercially valuable amino acid derivatives. However, microbial fermentation for 5-HTP production continues to face many challenges, including low titer/yield and the presence of the intermediate L-tryptophan (L-Trp), owing to the complexity and low activity of heterologous expression in prokaryotes. Therefore, there is a need to construct an efficient microbial cell factory for 5-HTP production. RESULTS: We describe the systematic modular engineering of wild-type Escherichia coli for the efficient fermentation of 5-HTP from glucose. First, a xylose-induced T7 RNA polymerase-PT7 promoter system was constructed to ensure the efficient expression of each key heterologous pathway in E. coli. Next, a new tryptophan hydroxylase mutant was used to construct an efficient tryptophan hydroxylation module, and the cofactor tetrahydrobiopterin synthesis and regeneration pathway was expressed in combination. The L-Trp synthesis module was constructed by modifying the key metabolic nodes of tryptophan biosynthesis, and the heterologous synthesis of 5-HTP was achieved. Finally, the NAD(P)H regeneration module was constructed by the moderate expression of the heterologous GDHesi pathway, which successfully reduced the surplus of the intermediate L-Trp. The final engineered strain HTP11 was able to produce 8.58 g/L 5-HTP in a 5-L bioreactor with a yield of 0.095 g/g glucose and a maximum real-time productivity of 0.48 g/L/h, the highest values reported by microbial fermentation. CONCLUSION: In this study, we demonstrate the successful design of a cell factory for high-level 5-HTP production, combined with simple processes that have potential for use in industrial applications in the future. Thus, this study provides a reference for the production of high-value amino acid derivatives using a systematic modular engineering strategy and a basis for an efficient engineered strain development of 5-HTP high-value derivatives.

Study on the nonfatigue and fatigue states of orchard workers based on electrocardiogram signal analysis.

In recent years, fatigue has become an important issue in modern life that cannot be ignored, especially in some special occupations. Agricultural workers are high-risk occupations that, under fatigue conditions over a long period, will cause health problems. In China, since very few studies have focused on the fatigue state of agricultural workers, we were interested in using electrocardiogram (ECG) signals to analyze the fatigue state of agricultural workers. Healthy agricultural workers were randomly recruited from hilly orchards in South China. Through the field experiment, 130 groups of 5-min interval ECG signals were collected, and we analyzed the ECG signal by HRV. The time domain (meanHR, meanRR, SDNN, RMSSD, SDSD, PNN20, PNN50 and CV), frequency domain (VLF percent, LF percent, HF percent, LF norm, HF norm and LF/HF) and nonlinear parameters (SD1, SD2, SD1/SD2 and sample entropy) were calculated and Spearman correlation coefficient analysis and Mann-Whitney U tests were performed on each parameter for further analysis. For all subjects, nine parameters were slightly correlated in nonfatigue and fatigue state. Six parameters were significantly increased and ten HRV parameters were significantly decreased compared the nonfatigue state. As for males, fifteen parameters were significantly different, and for females, eighteen parameters were significantly different. In addition, the probability density functions of SDNN, SDSD, VLF%, HFnorm and LF/HF were significantly different in nonfatigue and fatigue state for different genders, and the nonlinear parameters become more discrete compared the nonfatigue state. Finally, we obtained the most suitable parameters, which reflect the fatigue characteristics of orchard workers under different genders. The results have instructional significance for identifying fatigue in orchard workers and provide a convincing and valid reference for clinical diagnosis.

Chemosensory characteristics of regional Vidal icewines from China and Canada.

This work aimed to compare the flavor characteristics of Vidal icewines from China and Canada and to establish relationships between sensory descriptors and chemical composition. Descriptive analysis was performed with a trained panel to obtain the sensory profiles. Thirty important aroma-active compounds were quantified by four different methodologies. Partial least squares discriminant analysis was used to identify candidate compounds, which were unique to certain sensory descriptors. The sensory profiles of icewines from China were characterized by nut and honey aromas, while icewines from Canada expressed caramel and rose aromas. Nut and honey aromas had a close correlation with 1-hexanol, isoamyl acetate, phenethyl acetate and phenylethyl alcohol. Caramel aroma was correlated with ethyl esters and lactones and rose aroma was correlated with terpenes.