57 results about "Metabolism pathway" patented technology

The present invention is generally directed to the evolution of new metabolic pathways and the enhancement of bioprocessing through a process herein termed recursive sequence recombination. Recursive sequence recombination entails performing iterative cycles of recombination and screening or selection to “evolve” individual genes, whole plasmids or viruses, multigene clusters, or even whole genomes. Such techniques do not require the extensive analysis and computation required by conventional methods for metabolic engineering.

The invention discloses recombinant Escherichia coli producing salidroside, a construction method and applications thereof. The construction method comprises: (1) using Escherichia coli [delta]A as a starting strain, and carrying out gene knockdown or gene silencing to make the galE gene, the galT gene and the ugd gene on the Escherichia coli chromosome be not expressed to obtain an Escherichia coli strain BMGU; and (2) introducing genes such as ARO10, UGT73B6MK, pgm, galU and T7Polymerase to over-express the genes such as ARO10, UGT73B6MK, pgm, galU and T7Polymerase so as to achieve the heterologous synthesis of salidroside, such that the recombinant Escherichia coli producing salidroside is obtained. According to the present invention, the efficient UDP-glucosyltransferase mutant UGT73B6MK is introduced, and the glucose metabolism pathway is optimized, such that the yield of salidroside is significantly improved.

The invention discloses a method for treating high ammonia-nitrogen wastewater through nutrition conversion of mixotroph. The method for treating the high ammonia-nitrogen wastewater through nutrition conversion of the mixotroph comprises the following steps: 1, conducting transferring and culture on the mixotroph; 2, conducting high density culture in a culture medium rich in organic carbon or wastewater rich in organic carbon; 3, harvesting heterotrophic cells of the mixotroph; 4, transferring the heterotrophic cells into the high ammonia-nitrogen wastewater for autotrophic culture, absorbing high-concentration nitrogen and ammonia, and purifying the wastewater. According to the method, the method for changing the nutrition metabolism pathway of the mixotroph is introduced for treating the high ammonia-nitrogen wastewater, the treated wastewater can be recycled, the requirement of industrialized wastewater treatment conducted through microalgae is met, and the method is a new approach for conducting sewage treatment through produced microalgae economically and efficiently. The harvested microalgae cells can be further treated and used for preparing biological energy source, animal feed and the like.

According to the invention, the amino acid dehydrogenase activity of an L-valine fermentation strain is enhanced, and / or an Entner-Doudoroff (ED) metabolism pathway is activated, such that the problem of unbalanced reducing power during the L-valine fermentation process is solved, therefore the yield and the conversion rate of the L-valine produced by the escherichia coli are improved, and the one-step method anaerobic fermentation of the L-valine is achieved.

The invention discloses a gene NtAOS1 for improving jasmonic acid content in tobacco leaves, a cloning method and an application of the NtAOS1. The nucleotide sequence of the NtAOS1 which can improvethe content of jasmonic acid in tobacco is shown in SEQ ID NO. 1; the amino acid sequence encoding the protein is shown in SEQ ID NO. 2; a nucleic acid interference (RNAi) technology is used to designa segment of interference fragment, a gateway technology is used to construct plant expression vector and transform tobacco, and a qPCR technology is used to select NtAOS1 gene transcription level to significantly reduce transgenic plants. Detection of jasmonic acid content shows that the decreased expression level of the gene NtAOS1 results in significantly higher jasmonic acid content than that of wild-type control. The invention provides a material basis for applying the gene NtAOS1 and coded protein thereof to tobacco stress resistance breeding and subsequent research on jasmonic acid synthesis and metabolism pathway in tobacco.

The invention discloses a method for detecting tanshinol and its metabolites in a biological sample. The method is characterized by comprising the following steps that the biological sample is treated; a solution to be detected is detected by adopting an ultra-high performance liquid chromatography-mass spectrometer. By adopting the detecting method, more tanshinol metabolites in the biological sample can be detected, and accordingly a foundation is laid for illumination of tanshinol metabolism pathways.

The invention discloses a verbascoside biosynthesis pathway and verbascoside biosynthesis enzyme related genes. Tuberous roots of rehmannia at six different development stages serve as materials, overall RNA (ribonucleic acid) is extracted and detected, combined transcriptome sequencing is performed by an Illumina HiSeq2500 platform to obtain 149.8 million reads, and head assembly is performed to obtain 96961 Unigenes sequences. The Unigenes sequences are compared with a KEGG (Kyoto encyclopedia of genes and genomes) database to perform metabolism pathway analysis and obtain 280 KEGG pathways, a metabolism pathway is compared with a supposed verbascoside biosynthesis pathway, and 5 candidate KEGG pathways are screened. The supposed verbascoside biosynthesis pathway is compared with the screened 5 candidate KEGG pathways, 19 gene coding enzyme catalysis verbascoside biosynthesis pathways are built, the Unigenes sequences and the database are subjected to Blast X comparison, and ESTscan software functions are added to obtain 13 candidate verbascoside biosynthesis enzyme related genes. A foundation is laid for illuminating the complete verbascoside biosynthesis pathway and producing verbascoside by secondary metabolism engineering or synthetic biology.

The invention relates to a genetically engineered bacterium for synthesizing N-acetylglucosamine and an application of the genetically engineered bacterium. The genetically engineered bacterium is recombinant corynebacterium glutamicum containing a glucosamine synthetase gene glmS and a glucosamine transacetylase gene gna1, and is modified by a chassis microorganism of which related genes of a GlcNAc reverse transport pathway, a catabolism pathway and a competitive metabolism pathway are knocked out; the genetically engineered bacterium also contains a 6p-GlcNAc specific phosphatase gene yqaBused for increasing the concentration of extracellular GlcNAc, and the genetically engineered bacterium expresses the glucosamine synthetase gene glmS, the glucosamine transacetylase gene gna1 and the6p-GlcNAc specific phosphatase gene yqaB. The genetically engineered bacterium is safe and non-toxic, can produce the GlcNAc at high yield by utilizing a microbial fermentation method, and is stablein batch, relatively low in production cost and wide in application prospect.

The invention belongs to the field of biological energy sources and particularly relates to a nitrogen defect control method for production of ethanol through fermentation of synthesis gas with mixed bacteria. The method comprises steps as follows: a first stage: mixed bacteria are cultured to grow in a fermentation medium to reach a stable state, that is, the yield of fermentation products is stable and unchanged; a second stage: the mixed bacteria growing to reach the stable state are cultured at the second stage, the nitrogen concentration of the fermentation medium at the stage is controlled to be lower than that at the first stage, the mixed bacteria are the same type of acetogenic bacteria which come from cow dung, urban excess sludge and enriched products thereof, as ethanol is a non-growth coupling type product of the acetogenic bacteria, insufficient nitrogen nutrients possibly cause abnormal cell metabolism, cells enter a non-growth state or dormant state, the condition not beneficial to growth of the acetogenic bacteria is created through nitrogen nutrient defects, so that the metabolism pathway is changed from the acid producing phase to the solvent producing phase, and ethanol production is controlled.

The invention relates to a method for high flux synthesis of a nucleotide pool and assembling of a double-stranded DNA from semiconductor chips. The method comprises the following steps: 1, dividing a pre-synthesized double-stranded DNA into a plurality of DNA fragments; 2, designing nucleotides by using the DNA fragments; 3, arranging a flank sequence and a recombinase digestion identification sequence to each of two ends of each of the nucleotides to synthesize nucleotide sequences, wherein the nucleotide sequences are prepared from the semiconductor chips; 4, washing out the nucleotides from the semiconductor chips; 5, carrying out PCR amplification with the flank sequence as a primer to obtain a PCR product for later use, wherein the amount of every synthesized nucleotide obtained after the PCR amplification is greatly increased; and 6, splicing the nucleotides by using he recombinase digestion PCR product, and cutting while splicing to obtain double-stranded DNA fragments. The method has the advantages of low cost and high one-time synthesis flux, and can be used in assembling of metabolism pathways, biological approaches, biological element databases or genomes.

The invention belongs to the field of biomedicine and relates to application of a mevalonic acid metabolism pathway inhibitor and alphavirus in preparing antitumor drug. The invention finds for the first time that the mevalonic acid metabolism pathway inhibitor can be used for preparing an alphavirus antitumor synergist. The invention further relates to a drug composition and a drug set which respectively contain the mevalonic acid metabolism pathway inhibitor and the alphavirus and application of the inhibitor and the alphavirus in treating tumor, especially for tumor insensitive to the alphavirus.

The invention provides a method for detecting the content of linoleic acid in tissues by liquid chromatography-tandem mass spectrometry and a diagnostic kit of colorectal cancer. The method comprises the following steps: grinding a tissue sample into powder in liquid nitrogen; homogenizing the tissue powder with iced methanol containing BHT (Butylated Hydroxytoluene) and centrifuging; adding an interior label into supernatant and carrying out derivatization treatment with n-butanol; dispersing and dissolving the mixture by a mobile phase again to obtain a sample to be loaded and detected; analyzing the sample to be loaded and detected with the liquid chromatography-tandem mass spectrometry to obtain the content of the linoleic acid in the tissue sample. Furthermore, a metabolic enzyme of a linoleic acid pathway is subjected to expression analysis and a result shows that gene expression of FADS1 (Fatty Acid Desaturase 1), FADS2 (Fatty Acid Desaturase 2), ELOVL2 (elongase of very long chain fatty acid 2) and ELOVL5 (elongase of very long chain fatty acid 5) in the linoleic acid pathway is increased. The method provided by the invention can be used for detecting changes of the content of the linoleic acid of tumor tissues and normal tissues of patients with the colorectal cancer and expression changes of genes in a related metabolism pathway, and important reference is provided for diagnosis and treatment of the colorectal cancer and post-period nutrition support.

The invention discloses a metabonomics analysis method for a quorum sensing inhibition mechanism. Through extraction of bacterium metabolites and <1>H-NMR test, R language is adopted for carrying out orthogonal signal correction-partial least square method discrimination analysis on the <1>H-NMR spectrogram, screening out differentially expressed metabolites, calculating metabolite changing rates of a quorum sensing inhibition group and a control group, selecting the changed metabolites related to quorum sensing and carrying out metabolism pathway analysis, identifying the changed metabolism pathway and analyzing a bacterium quorum sensing inhibition mechanism of a quorum sensing inhibitor. The metabonomics analysis method disclosed by the invention has the advantages that metabonomics is adopted, the total metabolic state of an organism is dynamically and integrally described, and the action mechanism of the quorum sensing inhibitor can be effectively analyzed.

The invention discloses an application of a lactalbumin-oleic acid composite to preparation of medicines for restraining energy metabolism of tumor cells, and finds that alpha-LA-OA can resist the glycolysis process of tumor cells and reduce the consumption of oxygen by an aerobic respiration process of a mitochondrion and the generation amount of ATP. In addition, the invention further finds thatthe alpha-LA-OA can reduce generation of ATP in an HepG2 cell glucose metabolism way, so that the effect of restraining energy metabolism of tumor cells can be achieved; an HepG2 tumor cell model isused for confirming that the alpha-LA-OA can notably promote expression of mTORC2 target protein Rictor, mSin1 and Ser473-Akt, and the expression amount of various enzymes in the glycolysis way can benotably reduced. The alpha-LA-OA can activate an mTORC2 signal route, and can restrain activity of key enzymes in the glucose metabolism way, so as to reduce the energy metabolism for restraining tumor cells, by ATP.

The invention discloses application of a porphyrin pigment serving as immunologic adjuvant and further discloses application of the porphyrin pigment serving as a vaccine. The porphyrin pigment is a biological pigment, tissues outside an animal cardiovascular system can be combined with a pathogeny recognition receptor TLR4, and tissue damage and danger signals are simulated to build efficient stimulation to an immune system. The porphyrin pigment is also an ingredient of an animal and a human body, internal metabolic pathways are clear, uncertainty in the safety aspect does not exist, and the porphyrin pigment is suitable for preparation of the immunologic adjuvant and the vaccine for the animal, especially for human. The porphyrin pigment serving as the immunologic adjuvant can remarkably strengthen the humoral immunity effect of antigen or the vaccine and can reach the level of complete Freund adjuvant.

The invention relates to the use of benfotiamine (a synthetic derivative of vitamin B1) in preparing drugs for treating diabetic retinopathy, that is, a new clinical indication thereof. Benfotiamine can reduce the formation of 6-phosphofructose and 3-phosphoglyceraldehyde by activating the pentose phosphate pathway, thereby blocking abnormal metabolism pathways including DAG-PKC pathway, aminohexose pathway, polyalcohol pathway and AGEs formation pathway and preventing hyperglycemia-induced damage to retinal capillary endothelial cells. The compound is combined with pharmaceutically-acceptable adjuvant to prepare pharmaceutical preparations suitable for oral intake at a daily dose of 5-100mg, 1-3 times daily.

In the invention, a full-length blue-light induced bZIP type transcription factor (NgAUREO1) of 2080bp of nannochloropsis gaditana is cloned. The gene NgAUREO1 comprises an ORF of 1398bp, a 5'-end noncoding region of 192bp and a 3'-end noncoding region of 490bp and is capable of coding 465 amino acids. The gene NgAUREO1 comprises a bZIP structural domain and an LOV structural domain, wherein the bZIP structural domain can be combined with the promoter region of a specific gene to adjust and control the expression of the specific gene, and the LOV structural domain can sense the blue light of about 470nm, so that the gene NgAUREO1 has the characteristic of being capable of being adjusted and controlled by the blue light. The similarity between the sequence of the amino acid in the gene NgAUREO1 and the sequence of the amino acid in the gene aureochromel of vaucheria is 59%. A yeast expression vector of the gene NgAUREO1 is constructed, and the gene NgAUREO1 is introduced into saccharomyces cerevisiae to increase the content of oil in the yeast. The stimulation of the blue light to the recombinant saccharomyces cerevisiae proves that the blue light can serve as a switch to participate in adjusting and controlling the metabolism pathway of the fatty acid.

The invention discloses a prognosis analysis method based on primary liver cancer gene classification and liver cancer tissue energy metabolism. The prognosis analysis method comprises the following steps: acquiring clinical phenotype data, expression profile data and gene CNV and SNV mutation data of a liver cancer sample; calculating sample scores of four metabolic pathways of the tumor sample, and determining a plurality of optimal classifications; and analyzing differences of expression, gene mutation, clinical phenotypes and immune characteristics of a plurality of optimal classification metabolic pathways, and then comprehensively analyzing results to establish a primary liver cancer prognosis analysis model. The method has the advantages that the primary liver cancer gene classifier is constructed through a big data analysis technology, the molecular subtype of the primary liver cancer is determined, and the metabolic pathway expression difference, the gene mutation difference, the clinical manifestation difference and the immune difference of the molecular subtype of the primary liver cancer are analyzed, so that the primary liver cancer prognosis analysis model is established; the method is used for accurately predicting the prognosis of the primary liver cancer and provides a research basis for revealing an energy metabolism mode and a tumor microenvironment of the primary liver cancer.

The invention discloses fructokinase from a fungal traditional Chinese medicine and a coding gene and use thereof. The fungal traditional Chinese medicine fructokinase has an amino acid sequence as shown in SEQ ID NO: 1 and a nucleotide sequence of the coding gene as shown in SEQ ID NO: 2. The fungal traditional Chinese medicine fructokinase participates in synthesis of main precursor monosaccharide D-fructose of cordyceps polysaccharide from D-fructose-6-phosphoric acid. A fungal traditional Chinese medicine hirsutella sinensis polysaccharide biosynthetic metabolism pathway is studied in details in principle. A cloned DNA containing the nucleotide sequence can be transferred into engineering bacteria by transduction, transformation and conjugal transfer methods. An expression of a D-fructose biosynthetic gene is regulated, the host is endowed with high expressivity of D-fructose, an effective way is provided for increasing the yield of the D-fructose and a derivative thereof, and the fructokinase has a great application prospect.

The invention belongs to the technical field of biomass energy preparation, and particularly relates to a method for producing acetone-ethanol-butanol (ABE) by fermentation with a straw hydrolysate asa raw material. The method specifically includes the steps of straw pretreatment, biological enzymolysis, preparation of an ABE fermentation culture solution, a fermentation hydrolysate and the like.According to the method, when butanol is fermented with a straw acid hydrolysate as a raw material and in combination with the fermentation treatment of specific microbial strains, the expression level of related key enzymes in a solvent metabolism pathway can be significantly affected by adding glutathione, and thus chemical compositions of cell membranes of the strains are changed, and the butanol production capacity is improved. According to the method, straws are pretreated into hydrolysate and then directly fermented to produce ABE, the complex detoxification process is avoided, so thatthe production cost is reduced, and the industrialization of ABE fermentation is promoted; and furthermore, through the optimization of the fermentation process, especially the addition and utilization of the glutathione in the fermentation process, the fermentation yield of the ABE is further improved, and the ABE industrial production is promoted.

The invention relates to detection of drug metabolism in plasma, and in particular relates to a method for determining five ginsenosides in rat plasma, and the method is characterized in that: Ra1, Rb1, Rh1, Re and Ro in the plasma can be detected by a gas chromatography method by dichloromethane extraction after precipitating of proteins with methanol, and solid phase extraction column purifying.Compared with the prior art, the method has the advantages of high accuracy, high sensitivity and good repeatability, is suitable for simultaneous quantitative detection of the five ginsenosides in the plasma, and can be used for the study of ginsenoside metabolism pathways in the plasma.

The invention relates to application of polymerized hemoglobin in preparation of a medicine for preventing and treating respiratory failure. The medicine disclosed by the invention is an oxygen-carrying medicine and is also a colloidal solution. Compared with whole blood or red blood cells with an oxygen-carrying function, the red blood cell has the characteristics of good rheological property, long circulation retention time, normal physiological metabolic pathway and the like, and has the advantages of no need of matching, wide working temperature range (2222 DEG C), no blood coagulation risk, stable time of 36 months and the like compared with whole blood or red blood cells with an oxygen-carrying function. Wherein the particle size of polymerized hemoglobin is only one thousandth of that of red blood cells, but the oxygen carrying capacity is as high as 729 times of that of the red blood cells, and small-dose perfusion can relieve the tissue ischemia condition and relieve inflammatory response and can reach the microcirculation tail end which cannot be reached by the red blood cells to effectively supply oxygen. A neuroprotective agent can also be added to prepare a compound medicine, so that the inflammation state of a patient with respiratory failure can be reduced, and the damage of oxygen free radicals can be reduced.

The invention discloses a tryptanthrin derivative, and a preparation method and application thereof. The preparation method comprises the following steps: with 5-methylisatin as a raw material, oxidizing 5-methylisatin into 5-methylisatoic anhydride; under the weak alkaline condition of triethylamine, subjecting 5-methylisatoic anhydride and 5-fluoroisatin to reflux with acetonitrile as a solvent,and carrying out filtration and washing to obtain 2-methyl-8-fluorotryptanthrin; subjecting 2-methyl-8-fluorotryptanthrin and N-bromosuccinimide to reflux in a carbon tetrachloride solvent; and reacting a concentrated product with malonic acid in pyridine, and carrying out acidification, washing and filtering to obtain the desired tryptanthrin derivative. The tryptanthrin derivative of the invention has excellent inhibitory activity against indoleamine-2,3-dioxygenase (IDO), increases the water solubility of tryptanthrin, has broad application prospects and can be used for treating diseases with pathological features of the IDO-mediated tryptanthrin metabolism pathway, e.g., cancer, Alzheimer's disease, depression and cataract. The preparation method of the invention has the advantages ofsimple operation, mild conditions and the like, and is easy to be industrialized.

The invention discloses application of a porphyrin pigment serving as immunologic adjuvant and further discloses application of the porphyrin pigment serving as a vaccine. The porphyrin pigment is a biological pigment, tissues outside an animal cardiovascular system can be combined with a pathogeny recognition receptor TLR4, and tissue damage and danger signals are simulated to build efficient stimulation to an immune system. The porphyrin pigment is also an ingredient of an animal and a human body, internal metabolic pathways are clear, uncertainty in the safety aspect does not exist, and the porphyrin pigment is suitable for preparation of the immunologic adjuvant and the vaccine for the animal, especially for human. The porphyrin pigment serving as the immunologic adjuvant can remarkably strengthen the humoral immunity effect of antigen or the vaccine and can reach the level of complete Freund adjuvant.

The invention discloses an echinacea purpurea extract, which is prepared by the following steps: crushing echinacea purpurea, adding an ethanol solution with the volume ratio concentration of 20-50%, performing reflux extraction for more than two times, then combining extracting solutions, and performing concentrating first and then drying, thereby obtaining the echinacea purpurea extract. A preparation method of the echinacea purpurea extract and applications of the echinacea purpurea extract in the fields of regulating the intestinal content metabonomics of immunosuppressive rats and changing phosphoinositide metabolism, a phosphatidylinositol number system, ascorbic acid and aldose metabolism pathways of the immunosuppressive rats. The echinacea purpurea extract has the advantages that the echinacea purpurea extract has the effect of regulating immunosuppressive rat intestinal flora metabonomics, and animal experiments prove that the echinacea purpurea extract can improve the growth performance, and enhance the immune cell activity . The immune system is regulated by changing phosphoinositide metabolism, a phosphatidylinositol number system, ascorbic acid and aldose metabolism pathways.