BackgroundL-tryptophan is an aromatic amino acid widely used in the food, chemical and pharmaceutical industries. In Escherichia coli, L-tryptophan is synthesized from phosphoenolpyruvate and erythrose 4-phosphate by enzymes in the shikimate pathway and L-tryptophan branch pathway, while L-serine and phosphoribosylpyrophosphate are also involved in L-tryptophan synthesis. In order to construct a microbial strain for efficient L-tryptophan production from glucose, we developed a one step tryptophan attenuator inactivation and promoter swapping strategy for metabolic flux optimization after a base strain was obtained by overexpressing the tktA, mutated trpE and aroG genes and inactivating a series of competitive steps.ResultsThe engineered E. coli GPT1002 with tryptophan attenuator inactivation and tryptophan operon promoter substitution exhibited 1.67 ~ 9.29 times higher transcription of tryptophan operon genes than the control GPT1001. In addition, this strain accumulated 1.70 g l-1 L-tryptophan after 36 h batch cultivation in 300-mL shake flask. Bioreactor fermentation experiments showed that GPT1002 could produce 10.15 g l-1 L-tryptophan in 48 h.ConclusionsThe one step inactivating and promoter swapping is an efficient method for metabolic engineering. This method can also be applied in other bacteria.

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Polyhydroxybutyrate (PHB), the best known polyhydroxyalkanoates (PHA) has been believed to change intracellular metabolic flow and oxidation/reduction state, as well as enhance stress resistance of the host. In this study, a PHB biosynthesis pathway, which contains phaCAB operon genes from Ralstonia eutropha, was introduced into an L-tryptophan producing Escherichia coli strain GPT1002. The expression of the PHB biosynthesis genes resulted in PHB accumulation inside the cells and improved the L-tryptophan production. Quantitative real-time PCR analysis showed that the transcription of tryptophan operon genes in GPT2000 increased by 1.9 to 4.3 times compared with the control, indicating that PHB biosynthesis in engineered E. coli changed the physiological state of the host. Xylose was added into the medium as co-substrate to enhance the precursor supply for PHB biosynthesis. The addition of xylose improved both extracellular L-tryptophan production and intracellular PHB accumulation. Moreover, we obtained 14.4 g l(-1) L-tryptophan production and 9.7 % PHB (w/w) accumulation in GPT2000 via fed-batch cultivation.

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Escherichia coli grows fermentatively in glucose-containing medium under anaerobic condition with formation of a mixture of organic acids (lactate, acetate, formate, and succinate) and ethanol to accommodate reducing equivalents generated during glycolysis. In this paper, we tried to improve the lactate accumulation in E. coli by redox regulation genetically and physiologically. Our results indicated that genetic regulation of the host by reducing the reductive by-product may improve the lactate production. In addition, lactate accumulation was also improved under reduced and anaerobic cultivation conditions. Engineered E. coli SDU4 was able to accumulate lactate under strictly anaerobic conditions to 100 g/L with a yield of 1.97 mol/mol glucose.

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ABSTRACT:In recent years, the photorealistic 3D city models are increasingly important in various geospatial applications related to virtual city tourism, 3D GIS, urban planning, real-estate management. Besides the acquisition of high-precision 3D geometric data, texture reconstruction is also a crucial step for generating high-quality and visually realistic 3D models. However, most of the texture reconstruction approaches are probably leading to texture fragmentation and memory inefficiency. In this paper, we introduce an automatic framework of texture reconstruction to generate textures from oblique images for photorealistic visualization. Our approach include three major steps as follows: mesh parameterization, texture atlas generation and texture blending. Firstly, mesh parameterization procedure referring to mesh segmentation and mesh unfolding is performed to reduce geometric distortion in the process of mapping 2D texture to 3D model. Secondly, in the texture atlas generation step, the texture of each segmented region in texture domain is reconstructed from all visible images with exterior orientation and interior orientation parameters. Thirdly, to avoid color discontinuities at boundaries between texture regions, the final texture map is generated by blending texture maps from several corresponding images. We evaluated our texture reconstruction framework on a dataset of a city. The resulting mesh model can get textured by created texture without resampling. Experiment results show that our method can effectively mitigate the occurrence of texture fragmentation. It is demonstrated that the proposed framework is effective and useful for automatic texture reconstruction of 3D city model.  Corresponding Author