14 results about "Metabolic intermediate" patented technology

This invention discloses a method for the synergistic metabolic analysis of multiple components of traditional Chinese medicine (TCM) integrated with the gut microbiota. After initializing the microbiota ecosystem, a solution of multi-component TCM extracts is continuously injected into a microfluidic chip. Stable isotope-labeled precursors are pulsed into a specific simulation region to perform spatial isotope encoding. A standing wave acoustic field is applied at the sampling node of the microfluidic chip, and multiphase fluid spatial splitting is performed using the difference in the primary acoustic radiation force experienced by the particles to derive cell-free TCM metabolites. The cell-free TCM metabolites are separated by chromatography and gas phase drift separation. A four-dimensional metabolic feature array is constructed, and the absolute value of the mass shift is calculated for spatial inverse decoding. Closed-loop feedback is triggered based on the rate of change of target metabolite ion abundance, increasing the amplitude of the standing wave acoustic field pressure to confine intestinal bacteria and performing in-situ enrichment of synergistic metabolic intermediates. This invention realizes the spatial metabolic trajectory tracing and network reconstruction of multiple components of TCM in a complex gut microbiota ecosystem.

The invention belongs to the technical field of synthetic biology and enzyme engineering, and particularly relates to a multi-enzyme in-vivo space co-localization carbon metabolic flux strengthening method, a genetically engineered bacterium and application of the genetically engineered bacterium. According to the invention, a recombinant expression vector is constructed, key enzymes RHLA and RHLB participating in synthesis of a target product are respectively subjected to fusion expression with protein scaffold modules RIAD and RIDD through (G4S) 3 flexible connecting peptide, and are introduced into host chassis bacteria, so that a stable multi-enzyme composite system is formed in cells, the transmission distance of a metabolic intermediate is shortened, and the expression efficiency is improved. Spatial co-localization and concerted catalysis of key catalytic enzymes are realized, and the conversion efficiency of a carbon source to a target product is improved. Compared with the prior art, the genetically engineered bacterium constructed by the invention has the advantages of stable structure, no need of complex regulation, reasonable metabolic flow distribution and the like in the fermentation process, the synthesis efficiency and yield of the target biosurfactant are remarkably improved, and the feasibility of industrial amplification application is enhanced.

The invention relates to the technical field of gene engineering detection, and particularly discloses a gene detection system for promoting andrographolide synthesis based on a biosensor, a key gene ApDof29 for regulating and controlling andrographolide synthesis is determined through whole genome analysis, gene expression profile research and co-expression network analysis, the core regulation and control effect of ApDof29 is verified, and the gene detection system for promoting andrographolide synthesis based on the biosensor is used for promoting andrographolide synthesis. The invention also develops a real-time dynamic monitoring system, and realizes non-invasive monitoring of andrographolide content accumulation, biosynthesis rate and metabolic intermediate concentration change by utilizing the fusion of a specific response element and a reporter gene. Through comprehensive calculation processing of an accumulation abnormal coefficient, a synthesis rate abnormal coefficient and a concentration change abnormal coefficient, the regulation and control effect of ApDof29 is dynamically evaluated, and a scientific basis is provided for subsequent gene modification.

The invention provides a liquid chromatography-mass spectrometry method capable of efficiently detecting a deoxynivalenol (DON) metabolic intermediate product, namely 3-keto-DON (3-keto-DON). The detection method comprises a liquid phase elution procedure and a mass spectrum acquisition mode. According to the method, the detection time is 10 min, the retention time of 3-keto-DON is 6.99 min, the detection limit is 4 ng / ml, the quantitation limit is 10 ng / ml, the linear range is 10-1800 ng / ml, the correlation coefficient R2 is equal to 0.9995, and the relative standard deviation RSD is 1.99%.

This invention relates to a recombinant bacterial cell for de novo synthesis of xanthocyanin WGX-50 using glucose as a carbon source, its construction method, and its applications. The recombinant bacterial cell uses *Escherichia coli* as the host strain and integrates the following three heterologous biosynthetic modules: a 3,4-dimethoxyphenylethylamine de novo synthesis module, which contains an enzyme system catalyzing the generation of dopamine from L-tyrosine and further stepwise methylation of dopamine to 3,4-dimethoxyphenylethylamine; a cinnamoyl-CoA synthesis module, which contains an enzyme system catalyzing the generation of cinnamoyl-CoA from phenylalanine or glucose metabolic intermediates; and an amidation module, which contains an acyltransferase capable of catalyzing the condensation of 3,4-dimethoxyphenylethylamine and cinnamoyl-CoA to generate xanthocyanin WGX-50. This invention achieves, for the first time, the de novo biosynthesis of xanthocyanin in microorganisms, completely eliminating dependence on the expensive exogenous precursor 3,4-dimethoxyphenylethylamine and significantly reducing raw material costs.

Provided herein are methods and pharmaceutical compositions for preventing or reducing frequency or severity of convulsions or seizures, particularly epileptic seizures by administration of a therapeutically effective amount of 2-deoxy-D-glucose (2-DG) in combination with a therapeutically effective amount of beta-hydroxybutyrate, or 2-DG with other ketones or metabolic intermediates that increase brain ketones. Also provided are methods and pharmaceutical compositions for reducing severity and progression of traumatic brain injury and deleterious sequelae resulting therefrom in an animal by administration of a therapeutically effective amount of 2-DG in combination with a therapeutically effective amount of beta-hydroxybutyrate, or 2-DG with other ketones or metabolic intermediates that increase brain ketones.

The invention relates to the technical field of biology, in particular to a preparation method of fucosyltransferase and 2 '-fucosyllactose. The invention provides a fucosyltransferase, a nucleic acid molecule, a combined element, a host and a preparation method of 2 '-fucosyllactose. According to the invention, through knockout of a metabolic intermediate degradation gene and overexpression of a 2 '-fucosyllactose biosynthetic pathway, the yield of a 2'-fucosyllactose production engineering strain is greatly improved, and the economical efficiency of synthesis of 2 '-fucosyllactose by a fermentation method is improved.

The application provides an application of alpha-ketoglutaric acid in improving corneal endothelial injury and dysfunction, and belongs to the technical field of biological medicine. The application finds that, as a key metabolic intermediate in the tricarboxylic acid cycle, alpha-ketoglutaric acid can significantly reduce corneal endothelial cell injury and dysfunction induced by various causes such as high glucose, genetic defects, high intraocular pressure and surgical trauma by improving mitochondrial energy metabolism, reducing oxidative stress, stabilizing mitochondrial structure and membrane potential. Experiments prove that alpha-ketoglutaric acid can be used in the form of local administration such as eye drops, anterior chamber perfusion, systemic oral administration or combined administration, effectively reduces corneal edema, reduces central corneal thickness, shortens the recovery time of corneal transparency, and has a clear protection and repair effect on diabetic-related corneal endothelial lesions, Fuchs corneal endothelial dystrophy, glaucoma high intraocular pressure injury and intraocular surgery-related endothelial injury.

This invention provides a recombinant bacterium and its applications. After specific genetic engineering modifications, this recombinant bacterium can synthesize polyhydroxybutyrate (PHB) using one-carbon compounds such as methanol or carbon dioxide as a carbon source. Specifically, the metabolic pathway of the recombinant bacterium, after optimization, enables the efficient synthesis of PHB from methanol as a raw material via metabolic intermediates such as formaldehyde, 3-phosphoglyceric acid, and acetyl-CoA. The recombinant bacterium constructed in this invention can effectively utilize low-cost carbon sources such as methanol or carbon dioxide to produce PHB, and has promising prospects for industrial application.

A method for determining the amount of an analyte in a sample is described, wherein the analyte comprises insulin, proinsulin, a metabolic intermediate of proinsulin, a C-peptide, a mutant C-peptide, or a mixture of any two or more thereof. Provided herein are mass spectrometry methods for detecting and quantifying an analyte (e.g., insulin, proinsulins, metabolic intermediates of proinsulins, C-peptides, mutant C-peptides, or mixtures of any two or more thereof) in a biological sample using an enrichment and / or purification method in combination with mass spectrometry techniques.

The invention relates to a gene editing cell metabolic flow dynamic analysis method, system, equipment and medium, and the method comprises the following steps: carrying out mass spectrometry on a gene editing cell sample, identifying a metabolic intermediate in the gene editing cell sample, and obtaining dynamic change data; carrying out derivatization detection on the gene editing cell sample, and obtaining organic acid concentration data in a semi-quantitative manner; and processing the dynamic change data and the organic acid concentration data through a dynamic metabolism network model, and outputting metabolic flow dynamic data for quantitatively representing the state of a metabolic pathway, so as to achieve the purposes of dynamically and quantitatively analyzing the metabolic function of the gene editing cell and directly guiding the optimization of a gene editing strategy.

The invention relates to an organic acid derivatization reagent assisted synthetic route analysis method, system and equipment, and the method comprises the following steps: obtaining a fermentation broth sample of gene editing cells; carrying out chemical derivatization reaction on the intermediate metabolite by using an organic acid derivatization reagent, and converting the intermediate metabolite into a volatile derivative; introducing into a gas chromatograph-mass spectrometer, and separating and detecting the volatile derivative to obtain qualitative and quantitative data of an intermediate metabolite; analyzing metabolic flux distribution in the amino acid synthesis path through a metabolic flux dynamic analysis model; according to the metabolic flux distribution, identifying hidden metabolic nodes and flux limiting nodes in the amino acid synthesis path; and outputting analysis results of hidden metabolic nodes and flux limiting nodes to realize high-sensitivity capture and quantification of metabolic intermediates which are difficult to detect in gene editing cells, so that the aim of systematically analyzing a complete dynamic pathway of an amino acid synthesis network is fulfilled.

The application discloses application of 3-hydroxyglutaric acid in preparation of a medicine for preventing and / or treating a high uric acid related disease and a preventive and therapeutic medicine. The application firstly discovers and proves that 3-hydroxyglutaric acid has the activity of reducing the uric acid level in vivo, is a pioneering discovery of new use of known substances, and determines a "treatment window" which is lower than a toxicity threshold and can effectively reduce uric acid through a rigorous dose-effect relationship study. As a metabolic intermediate in vivo, the biological compatibility of 3-hydroxyglutaric acid at a low dose is expected to be better than that of a chemically synthesized medicine, and the potential side effect risk of long-term use is lower.

Provided herein are methods and pharmaceutical compositions for preventing or reducing frequency or severity of convulsions or seizures, particularly epileptic seizures by administration of a therapeutically effective amount of 2-deoxy-D-glucose (2-DG) in combination with a therapeutically effective amount of beta-hydroxybutyrate, or 2-DG with other ketones or metabolic intermediates that increase brain ketones. Also provided are methods and pharmaceutical compositions for reducing severity and progression of traumatic brain injury and deleterious sequelae resulting therefrom in an animal by administration of a therapeutically effective amount of 2-DG in combination with a therapeutically effective amount of beta-hydroxybutyrate, or 2-DG with other ketones or metabolic intermediates that increase brain ketones.