818 results about "Pyruvic acid" patented technology

The invention discloses construction and application of an engineering bacterium for co-expressing D-mannitol dehydrogenase and D-glucose dehydrogenase, and belongs to the technical field of genetic engineering. The technical problem to be solved by the invention is how to co-express D-mannitol dehydrogenase and D-glucose dehydrogenase through a biosynthetic pathway. In order to solve the technical problem, the invention provides recombinant escherichia coli. The recombinant escherichia coli is characterized by containing a D-mannitol dehydrogenase gene, a D-glucose dehydrogenase gene, a glucokinase gene, an acetyl coenzyme A synthetase gene, a D-lactose transporter gene, a D-glucose PTS permease-free gene and a pyruvate oxidase-free gene. And no regulator gene of galactose operon is contained. The invention establishes a preparation method for co-producing D-mannitol and sodium gluconate, which has guiding significance for the production of D-mannitol and sodium gluconate.

The invention relates to the technical field of microbial fermentation, and discloses a genetically engineered bacterium of high-yield D-pantothenic acid as well as a preparation method and application of the genetically engineered bacterium. The D-PA yield of the engineering bacteria is greatly improved through the following effects that acetolactate synthase IlvB and IlvN shunted by pyruvic acid are subjected to overexpression, meanwhile, a transcription attenuation region is deleted, and valine feedback inhibition sites are mutated; the method comprises the following steps: overexpressing keto-alcohol acid reductoisomerase IlvC and dihydroxy acid dehydratase IlvD, and converting the IlvC coenzyme preference from NADPH into NADH, so as to break through the upstream of D-PA synthesis; overexpression of PanB is carried out, overexpression of a serine-glycine transport system is carried out, and synthesis of 5, 10-methylenetetrahydrofolic acid is promoted; meanwhile, the expression of ketopantoic acid reductase PanE is enhanced; and finally, over-expressing pantothenic acid synthetase PanC, and increasing the content of a cofactor ATP required by the PanC. According to the engineering bacteria provided by the invention, through overall reinforcement of the multiple modules, cell growth is improved due to balance of cofactors and increase of D-PA, and the yield of D-PA reaches an unprecedented height.

The invention relates to the field of gene engineering, and discloses a D-3-phosphoglycerate dehydrogenase mutant, a coding gene thereof and application of the D-3-phosphoglycerate dehydrogenase mutant and the coding gene. Compared with a protein with an amino acid sequence as shown in SEQ ID NO: 1, the mutant comprises mutation at the 131 site and / or the 285 site. The mutant can specifically catalyze a substrate 3-phosphoglycerate to synthesize 3-phosphate-hydroxypyruvic acid, can effectively promote synthesis of L-serine in a metabolic pathway of microorganisms, improves the yield and conversion rate of the L-serine, and reduces the yield of byproducts at the same time.

The invention discloses construction and application of an L-malic acid producing issatchenkia orientalis strain, and belongs to the technical field of microorganisms and the technical field of microbial fermentation and metabolic engineering. Through a metabolic engineering means, a cytoplasm metabolic pathway of an issatchenkia orientalis strain is transformed, and a pyruvate decarboxylase coding gene, glycerol dehydrogenase and lactic dehydrogenase are knocked out; a pyruvate carboxylase gene derived from aspergillus flavus, a malate dehydrogenase gene derived from corynebacterium glutamicum and phosphoenolpyruvate carboxylase derived from escherichia coli are screened and integrated, so that a cytoplasm rTCA cycle synthesis pathway is enhanced; inserting a dicarboxylic acid external transporter gene from schizosaccharomyces cerevisiae at a neutral site GD21, and knocking out endogenous dicarboxylic acid internal transporter; the L-malic acid yield of the modified yeast strain in the fermentation process is remarkably increased and reaches 140.16 g / L. The engineering strain provided by the invention has good industrial application potential in efficient L-malic acid production.

The invention belongs to the technical field of chondroitin sulfate biosynthesis, and discloses a recombinant strain for high-yield chondroitin sulfate, application of the recombinant strain and a preparation method of the chondroitin sulfate. The recombinant strain provided by the invention can be used for simultaneously expressing UDP-glucosamine isomerase, UDP-glucose dehydrogenase, chondroitin polymerase, glucosamine synthetase, phosphoglucosamine mutase, UDP-N-acetylglucosamine pyrophosphorylase, glucose-6-phosphate isomerase and glucose phosphate mutase; according to the present invention, glucose-1-phosphate uridyl transferase, ATP sulfonase, sulfotransferase, pyruvate kinase and APS kinase are adopted, and the enzyme molecules are matched with the original carbon metabolism pathway in the recombinant strain cell to jointly form the enzyme-linked pathway having the efficient CSA synthesis ability, such that the high-yield and high-purity preparation of the CSA can be achieved; the method has the advantages of simple operation, high biological safety of the obtained product, easy realization of large-scale production and the like, and has a good application prospect.

The invention provides recombinant escherichia coli for de novo synthesis of anisic acid. At least one of the following improvements is carried out on an escherichia coli starting strain: (1) overexpression of O-methyltransferase PSA-OMT1 from Pleuromyces sapidus, (2) overexpression of O-methyltransferase PSA-OMT1 from Pleuromyces sapidus, and (3) overexpression of O-methyltransferase PSA-OMT1 from Pleuromyces sapidus; the method comprises the following steps: carrying out overexpression on chorismate lyase UbiC derived from E.coli MG1655 or Providencia rustiana, and carrying out the overexpression on the chorismate lyase UbiC derived from E.coli MG1655 or Providencia rustiana; (2) carrying out overexpression on a 3-deoxy-D-arabic-heptanone acid 7-phosphate (DAHP) synthase feedback inhibition mutant AroGD146N derived from E.coli MG1655 on the basis of the step (1); and (3) overexpressing synthase PpsA derived from phosphoenolpyruvate (PEP) on the basis of the step (2), and knocking out a pykF gene for coding pyruvate kinase I. Therefore, in the fermentation process of the recombinant escherichia coli, the accumulation of the anisic acid in the fermentation liquor can be realized, the maximum anisic acid shake flask yield of the recombinant escherichia coli can reach 284.9 mg / L, and a foundation is laid for the subsequent construction of high-yield anisic acid engineering bacteria.

The invention provides a porcine oocyte in-vitro maturation culture solution additive and application thereof, and belongs to the field of embryo engineering, and the additive contains taurine. The additive also contains sodium pyruvate, cysteine, glucose, NaHCO3, fetal calf serum, estradiol, pregnant horse serum gonadotropin and human chorionic gonadotropin. When the porcine oocyte in-vitro maturation culture solution added with the additive is used for culturing porcine oocytes in vitro, the ROS content in the porcine oocytes can be remarkably reduced, the oxidation resistance of the porcine oocytes can be improved, and the mitochondrial distribution of the oocytes can be improved, so that the development of the oocytes can be improved; the maturation of porcine oocytes and the development of early embryos are promoted. When the technology is applied to the pig assisted reproduction technology, the success rate and quality of embryo engineering can be improved; the pig oocyte in-vitro maturation culture solution added with the additive can improve the premature embryo division rate in parthenogenetic activation of the pig oocytes.

The invention belongs to the technical field of biological medicines, and particularly relates to a pyruvate preparation for treating asthma and a preparation method of the pyruvate preparation. The pyruvate preparation is prepared from the following components in parts by weight: 10 to 12 parts of pyruvate, 10 to 12 parts of sodium chloride, 0.5 to 1 part of citric acid and 500 parts of water for injection. The pH value of the pyruvate preparation is 6 to 7. The pyruvate preparation is used for daily management of asthma patients, the purpose of inhibiting asthma is achieved by controlling increase of the number of inflammatory cells such as eosinophilic granulocytes and reducing inflammatory reaction, the effect of the pyruvate preparation is close to that of similar hormone medicine dexamethasone, and the effect of the pyruvate preparation is improved while the treatment purpose is achieved. The dependence and side effects of patients on hormone drugs are reduced.

The invention belongs to the technical field of medicines, and particularly relates to a pyruvate-containing composite preparation as well as a preparation method and application thereof. The composite preparation containing pyruvate is prepared from the following components in percentage by mass: 0.02 to 0.04 percent of a compound A, 0.05 to 0.1 percent of pyruvate, 0.2 to 2 percent of sodium dihydrogen phosphate, 0.2 to 2 percent of disodium hydrogen phosphate, 0.7 to 0.9 percent of sodium chloride and the balance of water for injection, the structural formula of the compound A is as follows: # imgabs0 #. The compound A and pyruvate in the compound preparation interact with each other, so that lung inflammatory response can be inhibited, the level of inflammatory factors can be reduced, and lung edema can be relieved.

The present disclosure provides a recombinant yeast cell for making ethanol. The recombinant yeast cell comprises a first genetic modification to increase an ethanol yield in the recombinant yeast cell when compared to a parental yeast cell. The recombinant yeast cell also comprises a second genetic modification capable of increasing pyruvate decarboxylase activity in the recombinant yeast cell when compared to the parental yeast cell. The parental yeast cell lacks the first genetic modification and the second genetic modification. The present disclosure also provides methods for making the recombinant yeast cell as well processes for using the recombinant yeast cell to make ethanol.

The invention relates to the field of biological medicines, in particular to a composition containing pyruvate as well as a preparation method and application of the composition. The composition containing pyruvate is prepared from the following raw materials in percentage by mass: 0.05 to 0.1 percent of pyruvate, 0.02 to 0.04 percent of bergenin derivative, 5 to 10 percent of water for injection and the balance of propellant, the structural formula of the bergenin derivative is as follows: # imgabs0 #. The composition containing pyruvate can reduce the abundance of pathogenic bacteria and increase the abundance of probiotics, so that the effect of treating lung diseases is achieved by adjusting the respiratory tract micro-ecology.

The present disclosure relates to compounds that are capable of inhibiting the mitochondrial pyruvate carrier and promoting hair growth. The disclosure further relates to methods of promoting hair growth or treating conditions or disorders affecting hair growth, such as baldness or alopecia.

A transformant produced by introducing a chorismate pyruvate-lyase gene comprising DNA (a), (b), (c), (d) or (e) and a 4-hydroxybenzoate hydroxylase gene comprising DNA (f), (g), (h), (i) or (j) into a Hydrogenophilus bacterium can utilize carbon dioxide as a sole carbon source to produce protocatechuic acid with high efficiency. (a) DNA that comprises the nucleotide sequence represented by SEQ ID NO:1; (b) DNA that comprises a nucleotide sequence having 90% or higher identity with SEQ ID NO:1 and encodes a polypeptide having a chorismate pyruvate-lyase activity; (c) DNA that encodes a polypeptide comprising the amino acid sequence represented by SEQ ID NO:2; (d) DNA that encodes a polypeptide comprising an amino acid sequence having 90% or higher identity with SEQ ID NO:2 and having a chorismate pyruvate-lyase activity; (e) DNA that encodes a polypeptide comprising an amino acid sequence having the deletion, substitution, insertion or addition of 1 to 10 amino acid residues in the amino acid sequence represented by SEQ ID NO:2 and having a chorismate pyruvate-lyase activity; (f) DNA that comprises the nucleotide sequence represented by SEQ ID NO:3; (g) DNA that comprises a nucleotide sequence having 90% or higher identity with SEQ ID NO:3 and encodes a polypeptide having a 4-hydroxybenzoate hydroxylase activity; (h) DNA that encodes a polypeptide comprising the amino acid sequence represented by SEQ ID NO:4; (i) DNA that encodes a polypeptide comprising an amino acid sequence having 90% or higher identity with SEQ ID NO:4 and having a chorismate pyruvate-lyase activity; and (j) DNA that encodes a polypeptide comprising an amino acid sequence having the deletion, substitution, insertion or addition of 1 to 30 amino acid residues in the amino acid sequence represented by SEQ ID NO:4 and having a 4-hydroxybenzoate hydroxylase activity.

The invention relates to a method for increasing the yield of L-malic acid produced by fermentation by adding ethylene glycol, which comprises the following steps: constructing genetic engineering escherichia coli, and introducing ethylene glycol metabolism genes (overexpression) including glycerol dehydrogenase, lactaldehyde reductase, lactaldehyde dehydrogenase and the like; and overexpression of other genes involving malic acid production, such as malic acid synthase, phosphoenolpyruvate carboxylase and malic acid export protein. The gene engineering strain is an E.coli BW25113 strain of which ldhA, pta, poxB, adhE, pflB, mgsA, frdA, maeA, maeB, yqhD, ycdW and iclR genes are substantially inactivated, and the deleted genes are beneficial to reducing the generation of metabolic byproducts and increasing the accumulation of malic acid. The genetic engineering strain is subjected to fermentation culture by taking glucose and ethylene glycol as carbon sources, and L-malic acid is separated from a culture after culture is finished.

The invention provides a method for increasing the yield of bacterial cellulose by utilizing low-dose ultraviolet radiation, and belongs to the field of microbial fermentation. The method comprises the following steps: inoculating a fermentation culture medium with a Taonella.mepensis seed solution, carrying out fermentation treatment, and carrying out ultraviolet radiation under set conditions in the fermentation process, so as to obtain a fermentation solution containing a bacterial cellulose membrane; and taking out the bacterial cellulose membrane in the fermentation liquor, and purifying the bacterial cellulose membrane to obtain the bacterial cellulose. On the basis of an HS culture medium formula, the ultraviolet irradiation time is regulated and controlled at the same time, so that the activity of cellulose synthesis related enzymes, such as the activity of UDP-glucose pyrophosphorylase and glucose phosphate mutase, is improved, the activity of glucose decomposition pathway related enzymes, such as pyruvate kinase and phosphofructokinase, is inhibited, and the activity of cellulose synthesis related enzymes is improved. The yield of the bacterial cellulose is further improved.

The invention discloses a serum-free culture medium for in-vitro proliferation of poultry muscle stem cells and application of the serum-free culture medium, and belongs to the technical field of cell engineering, a basic serum-free culture medium is DMEM-F12, a complete serum culture medium is a mixture of DMEM-F12 added with 10% of fetal calf serum and 1% of mycillin, and the volume ratio of a culture medium supplement to the basic culture medium is 1: 1000; the serum-free culture medium contains a supplement; the supplement comprises the following compositions: insulin, transferrin, fibroblast growth factor 2, epidermal growth factor, catalase, ethanolamine, vitamin B12, linoleic acid, linolenic acid, retinol acetate, tocopherol, L-alanine, L-proline, sodium selenite, sodium pyruvate, reduced glutathione, berberine hydrochloride and analogues thereof.

The invention relates to a repetitive structural unit recombinant protein cpE2 and a preparation method and application thereof. The recombinant protein cpE2 of the repetitive structural unit has self-assembly performance and is formed by modifying an E2 subunit of a pyruvate dehydrogenase compound and dihydrofatty acid amide transacetylase. Specifically, the C end of the E2 protein is arranged at the N end through a connecting peptide by utilizing a circulating recombination principle, so that the recombinant protein cpE2 is constructed. The engineered cpE2 protein cage can be self-assembled into a cage-shaped structure, linking groups can be respectively brought into the protein cage through N-terminal or C-terminal modification of cpE2, the problem that a modifier or a drug is difficult to put into the protein cage by a traditional E2 protein is solved, meanwhile, the specific particle size of the cpE2 protein cage can be controlled through the length of a linking peptide, and the protein cage has a good application prospect. And efficient carrying of goods is achieved.

The present invention relates to the use of pyruvate dehydrogenase kinase (PDK4) inhibitor sodium dichloroacetate (DCA) in the treatment of heart failure with preserved ejection fraction. The use specifically comprises: the pyruvate dehydrogenase kinase (PDK4) inhibitor sodium dichloroacetate (DCA) is administrated orally to prevent and / or treat heart failure with preserved ejection fraction.

The invention belongs to the technical field of biological medicines, and particularly relates to sodium pyruvate sustained-release gel nanoparticles based on redox response as well as a preparation method and application of the sodium pyruvate sustained-release gel nanoparticles. The preparation method comprises the following steps: adding modified gellan gum containing sulfydryl and catechol groups, phenylboronic acid modified gellan gum and sodium pyruvate into water to obtain a mixed solution; under the protection of inert gas, divalent metal salt and potassium persulfate are added into the mixed solution, stirring is performed for 3-5 h, after the reaction is finished, centrifugation is performed, precipitates are collected, washed and dried, and the composite material is obtained. Experimental results show that the sodium pyruvate sustained-release gel nanoparticles prepared by the invention can realize controlled release of drugs, reduce toxic and side effects caused by burst release of the drugs, improve the curative effect of the drugs and improve the bioavailability.

The invention discloses escherichia coli recombinant bacteria with high glycolic acid yield and a construction method of the escherichia coli recombinant bacteria, and belongs to the technical field of bioengineering. Lactate dehydrogenase ldhA, phosphate transacetylase pta, acetokinase ackA and pyruvate oxidase poxB are knocked out in escherichia coli, key enzymes isocitrate lyase aceA and glyoxylate reductase ghrA in a glycollic acid synthesis path are over-expressed in a genome level, conversion of carbon flow to glycollic acid is enhanced, and the glycolic acid is converted into glycolic acid. On the basis, malic acid synthase AaceB, malic acid synthase GglcB and glycollic acid oxidase glcDEF are knocked out, a branch metabolic pathway is blocked, an efficient strain which is free of plasmids and resistance genes and does not need to induce glycollic acid production is constructed, the accumulation amount of L-valine fermented in a 5L fermentation tank for 48 hours reaches 51.2 g / L, the yield is 0.6 g / g, and the production intensity is 1.06 g / L / h.

The invention provides a construction method and application of recombinant yarrowia lipolytica with high yield of tyrosol, and belongs to the field of bioengineering. The recombinant yarrowia lipolytica with high yield of tyrosol is named as a yarrowia lipolytica XJ-TL strain, and the recombinant yarrowia lipolytica with high yield of tyrosol is named as a yarrowia lipolytica XJ-TL strain. The recombinant yarrowia lipolytica disclosed by the invention expresses an ethanol dehydrogenase encoding gene, and simultaneously expresses any one or a combination of more of a DAHP synthase mutant encoding gene, a chorismate mutase mutant encoding gene, a glucose 6-phosphate dehydrogenase encoding gene, a transketolase encoding gene and a transaldolase encoding gene; meanwhile, a natural promoter of a pyruvate kinase coding gene is truncated. After the recombinant yarrowia lipolytica XJ-TL strain is subjected to fed-batch fermentation, tyrosol can be synthesized at a relatively high level. The construction method of the recombinant yarrowia lipolytica is easy to operate, and efficient synthesis of tyrosol can be achieved.

The invention relates to the technical field of microbial metabolism engineering, in particular to a corynebacterium glutamicum producing strain for efficiently producing D-pantothenic acid as well as a construction method and application of the corynebacterium glutamicum producing strain. The construction method specifically comprises the following steps: (1) by taking wild corynebacterium glutamicum ATCC 13032 as a chassis, weakening ilvA, and constructing DPA1 through mutation expression; (2) knocking out the quinone pyruvate oxidoreductase and inserting a panBCE gene expression cassette into the quinone pyruvate oxidoreductase to construct DPA2; (3) introducing and using a pEC-xk99E overexpression plasmid to screen and express Escherichia coli W3110, Bacillus subtilis 168, and endogenous acetohydroxy acid synthase, acetolactate isomerized reductoisomerase and 2-hydroxy acid dehydratase, and carrying out a step (2) of carrying out a step (3) of screening and expressing the Escherichia coli W3110, Bacillus subtilis 168, and endogenous acetohydroxy acid synthase, acetolactate isomerized reductoisomerase and 2-hydroxy acid dehydratase in the DPA2; and (4) under the control of a strong promoter, carrying out tandem overexpression on the screened ilvBNCD gene by using pEC-xk99E, constructing DPA3, and enhancing a pantothenic acid main synthesis pathway and an upstream pathway pulling force, thereby obtaining the corynebacterium glutamicum for efficiently producing D-pantothenic acid. The strain provided by the invention can produce D-pantothenic acid with high yield and high conversion rate by utilizing a carbon source, and has important industrial application value.

The invention belongs to the field of metabolic engineering, and discloses a genetically engineered bacterium of high-yield beta-alanine as well as a construction method and application of the genetically engineered bacterium. The genetically engineered bacterium takes escherichia coli W3110 as an original strain, and is obtained by performing the following gene editing on a genome of the escherichia coli W3110, the genetically engineered bacterium does not express proteins such as lactose operon repressor protein, pyruvate dehydrogenase, acetokinase and the like; protein such as aspartic acid-alpha-decarboxylase, phosphoenolpyruvate carboxylase, pyruvate carboxylase and the like is overexpressed; the phosphoenolpyruvate carboxykinase is replaced by phosphoenolpyruvate carboxykinase from Mannheim succinic acid producing bacteria. The genetically engineered bacterium BA-17 for high yield of beta-alanine, which is constructed by the invention, is fermented in a 5L fermentation tank for 60 hours, the yield of beta-alanine reaches 126.7 g / L, the conversion rate reaches 60.3%, and the production intensity reaches 2.11 g / L / h.

The invention discloses pyruvate carboxylase (PC) serving as a biomarker for detecting endometriosis and application of the pyruvate carboxylase (PC) to treatment of the endometriosis. The biomarker can be used for detecting the expression level of PC in an endometrial tissue sample through an immunohistochemical method, and when the IHC score is greater than or equal to 4, a subject is judged to be at a high risk of endometriosis or suffered from the endometriosis. Meanwhile, the invention provides an application of drugs (including specific siRNA, ZY-444, octyl gallate and the like) for inhibiting PC expression in treatment of endometriosis, and a corresponding treatment method. The invention provides a new biomarker, target spot and scheme for detection and treatment of endometriosis, and has important clinical application value.

The invention discloses a method for catalytic synthesis of malonic acid by using whole cells, and belongs to the field of bioengineering. According to the invention, escherichia coli BL21 (DE3) is taken as a host, an aspartic acid ammonia lyase gene aspA and an aspartic acid-alpha-decarboxylase gene panD, a succinate semialdehyde dehydrogenase gene yneI and a beta-alanine pyruvate transaminase gene, a beta-alanine pyruvate transaminase gene bauA and a malonyl-coenzyme A reductase gene mcr-C are expressed in modules, and recombinant escherichia coli is constructed. The malonic acid is produced by adopting a whole-cell catalysis method, so that the production period is shortened, the production efficiency of the malonic acid is improved, and the yield of the malonic acid can reach 51.0 g / L after the malonic acid is catalyzed for 24 hours.

The invention provides a valine production strain and a construction method and application thereof, a designed acetohydroxy acid synthase mutant is that the 88th basic group of an ilvB gene is changed from a to c, the 382nd basic group is changed from a to g, the 413th basic group is changed from c to t, the gene sequence of a designed artificial operon comprises a promoter, an ilvB (A138V) gene or ilvB (Q30K, S128G, A138V) gene of coding mutated acetohydroxy acid synthase, and an ilvN (G20D, I21D, I21D, I21D, I21D, I21D, I21D, I21D) gene. I22F) gene, a pyk gene for coding pyruvate kinase, and a terminator; by designing a specific acetohydroxyacid synthase mutant and related biological materials and artificial operon, the strain constructed by directional modification of the strain by using a pK18mobsacB system gene editing technology based on allele exchange has the advantages of good genetic stability, high fermentation yield and the like, and valine can be stably produced.

The invention discloses an application of cinobufagin in preparation of a medicine for inhibiting bone metastasis of lung cancer. It is proved for the first time that 1) in-vitro cell experiments prove that cinobufagin can effectively inhibit proliferation and migration ability of lung cancer bone metastasis cells and promote cell apoptosis; 2) a mouse left ventricular injection bone metastasis model and a tibia injection model show that cinobufagin can significantly inhibit the development of lung cancer bone metastasis and osteolytic bone injury; (3) the specific regulation and control mechanism of cinobufagin for inhibiting bone metastasis of lung cancer is explored, and it is found through combined analysis of IP-MS proteomics and LiP-SMap proteomics that cinobufagin may target pyruvate carboxylase to play the function of cinobufagin; and 4) further verifying that cinobufagin can be combined with PC to inhibit bone metastasis of lung cancer through a drug affinity reactive target stability experiment and a heat transfer experiment. The research provides a new target spot and a treatment strategy for inhibiting bone metastasis of lung cancer.

The invention discloses a tobacco leaf surface spraying composition and application thereof, and belongs to the technical field of organic fertilizers. The tobacco leaf surface spraying composition is prepared from the following components in percentage by weight: 0.50 percent to 1.00 percent of lycopene, 0.06 percent to 0.10 percent of phosphoenolpyruvic acid and the balance of water. According to the method, the lycopene and the phosphoenolpyruvic acid with specific contents are sprayed on the leaf surfaces of the tobacco leaves, so that the content of key volatile metabolites closely related to the sensory quality of the flue-cured tobacco in the fresh and sweet flavor type flue-cured tobacco can be increased, the sensory quality of the fresh and sweet flavor type flue-cured tobacco is improved, and the fresh and sweet flavor type of the flue-cured tobacco is enhanced.

The invention discloses a production strain for fermenting L-histidine and application of the production strain, and belongs to the technical field of biological engineering. According to the invention, escherichia coli E.coli CICC 10243 is taken as an original strain, histidine synthesis operon genes hisDCB-cg2302-cg2301-HA-impA-FI, hisN and hisEG from corynebacterium glutamicum are integrated on a genome of the escherichia coli E.coli CICC 10243, a pyruvate dehydrogenase AceE mutant coding gene is introduced, and an escherichia coli transcription factor SlyA coding gene slyA is inactivated. The novel strain E.coli His08 for fermentation production of L-histidine is prepared and fermented in a 5L fermentation tank for 60 h, the histidine yield reaches 58.39 g / L, the sugar-acid conversion rate is 0.15 g / g, and the strain E.coli His08 has important application value.

The invention relates to a novel hexahydropyrroloindole alkaloid extracted from fructus lycii, which can effectively solve the problems of extracting and preparing the novel hexahydropyrroloindole alkaloid A from the fructus lycii and realizing the application of the novel hexahydropyrroloindole alkaloid A in preparation of medicines for preventing and treating type 2 diabetes mellitus. The novel hexahydropyrroloindole alkaloid A is prepared by the following steps: sequentially eluting with water, ethanol and acetone, carrying out reduced pressure concentration on an ethanol part, extracting, stirring a sample with silica gel, carrying out gradient elution, merging the same fractions, eluting a target component, stirring the sample with silica gel, carrying out gradient elution, collecting fractions within retention time, concentrating and drying, thereby obtaining the novel hexahydropyrroloindole alkaloid A in the preparation of the medicine for preventing and treating the type 2 diabetes mellitus and the application of the novel hexahydropyrroloindole alkaloid A in the preparation of the medicine for preventing and treating the type 2 diabetes mellitus. Glucose consumption in insulin-resistant HepG2 cells can be remarkably promoted, glucose uptake capacity is improved, glycogen synthesis is promoted, activity of glycolysis key enzymes hexokinase and pyruvate kinase is enhanced, development and application prospects are wide, and economic and social benefits are remarkable.