33 results about "Deoxynojirimycine" patented technology

The invention discloses a preparation method and application of a mulberry leaf extract with high stability and high activity in a protein food system, and belongs to the field of food processing and nutrition and health. The mulberry leaf extract is prepared by taking mulberry leaves as a raw material through steam explosion and ball milling pretreatment, compound enzyme directional enzymolysis, ultrasonic-assisted extraction, protein-induced sedimentation combined with ultrafiltration membrane selective impurity removal, compound stabilization treatment and drying. The obtained extract is rich in 1-deoxynojirimycin (DNJ) and mulberry leaf polysaccharide, high-polymerization-degree polyphenol is remarkably removed, and high clarity and heat stability are achieved. The DNJ retention rate gt is obtained after the milk is stored at 37 DEG C for 7 days when the milk protein is applied to a milk system; and therefore, the product prepared by the method is obviously superior to the product prepared by the prior art. The extract is pure in flavor and high in biological activity, and can be widely applied to the field of healthy foods such as dairy products and protein beverages.

The invention relates to the technical field of compound extraction and separation, and discloses a method for extracting and separating 1-deoxynojirimycin from mulberry leaves, which comprises the following steps: S1, mulberry leaf pretreatment: crushing and sieving mulberry leaves to obtain mulberry leaf powder; s2, extracting: extracting mulberry leaf powder by adopting a mixed solvent of water and ethanol, repeatedly extracting twice, and combining extracting solutions; s3, preliminary impurity removal; s4, chromatographic purification: S401, enabling the supernate to pass through a strong acid cation exchange resin column, eluting with deionized water, then eluting with an ammonia water solution, collecting eluent, and adjusting the eluent to be neutral to obtain neutral feed liquid; s402, enabling the neutral feed liquid to pass through a macroporous adsorption resin column, eluting with deionized water, then eluting with an ethanol solution, and collecting a target eluent containing 1-deoxynojirimycin; and S5, post-processing. Adsorption and separation of DNJ are sequentially carried out through a two-step chromatography process of strong acid cation exchange resin and macroporous adsorption resin, and various impurities are effectively removed.

The invention belongs to the technical field of biology, and discloses a transaminase GabT1 mutant S106I, a construction method thereof and application of the transaminase GabT1 mutant S106I in biosynthesis of 1-deoxynojirimycin. According to the invention, the 106th serine near the molecular catalytic domain of the transaminase GabT1 derived from bacillus amyloliquefaciens is mutated into isoleucine in a site-directed mutagenesis manner, so that the catalytic efficiency of the transaminase GabT1 is remarkably improved, the problem that the catalytic efficiency of the transaminase GabT1 on fructose-6-phosphoric acid is not high at present is solved, and the application of the transaminase GabT1 to the fructose-6-phosphoric acid is promoted. The yield of 1-deoxynojirimycin synthesized by the mutant GabTS106I is increased by 43.87% compared with that of a wild type. The 1-deoxynojirimycin fermentation production capability of the transaminase GabT1 mutant and the engineering strain obtained by the invention is greatly improved, and the transaminase GabT1 mutant and the engineering strain have a good industrial application prospect.

The invention discloses a phenylboronic acid affinity deoxynojirimycin imprinted polymer nano-material as well as a preparation method and application thereof, and belongs to the fields of nano-material preparation and natural compound extraction and separation. Phenylboronic acid modified nano silica gel is used as a carrier, deoxynojirimycin is used as a template molecule, dopamine is used as a cross-linking agent, a polyethyleneimine hyperbranched strategy is further combined, a deoxynojirimycin molecularly imprinted polymer (MIPs-PEI) with boric acid affinity and pH response characteristics is constructed, and hyperbranched modification remarkably improves hydrophilicity and boric acid affinity sites of the material. The synthesized MIPs-PEI has good adsorption performance on deoxynojirimycin and can effectively enrich deoxynojirimycin in mulberry leaves, and the natural hypoglycemic alkaloid deoxynojirimycin is extracted in a green mode through the method.

The invention provides a method for purifying 1-deoxynojirimycin in the root bark of white mulberry. The method comprises the following steps: adding white mulberry root-bark powder into a deep eutectic solution, heating and extracting, and filtering to obtain an extracting solution; adding 0.3%-1% of a novel protein precipitant into the extracting solution, precipitating and filtering to obtain a column loading solution; the upper injection liquid passes through a pretreated cationic resin column, washing is performed after adsorption, ammonia water elution is performed, and the eluent is dried to obtain 1-deoxynojirimycin; wherein the deep eutectic solution comprises the following components in percentage by weight: 0.1%-0.5% of choline chloride, 0.1%-1.5% of citric acid and water; the preparation method of the novel protein precipitant comprises the following steps: dissolving chitosan in a citric acid solution, heating and emulsifying, adding a demulsifier for demulsification after emulsification is completed, and filtering to obtain the novel protein precipitant. According to the method, through the treatment method that deep eutectic solvent extraction is combined with protein precipitant treatment and then column purification is conducted, the extraction efficiency of DNJ can be effectively improved, and protein impurities can be better removed.

The present invention belongs to the field of industrial biotechnology, and discloses a method for improving 1-deoxynojirimycin bioproduction strains, ybbA gene and application, the method comprising introducing the ybbA gene, an expression cassette comprising the ybbA gene or a vector comprising the ybbA gene into Bacillus amyloliquefaciens. The present invention uses precise gene knockout and backfilling technology to conduct a comprehensive functional analysis of the ybbA gene in Bacillus amyloliquefaciens. It aims to clarify the exact role of the ybbA gene in DNJ biosynthesis and provide a scientific basis and theoretical support for the development of novel and efficient DNJ production strategies. The research results of the present invention are expected to break through the bottleneck of DNJ yield improvement and open up new paths and methods for related research in the field of industrial biotechnology.

The invention discloses a method for co-producing mulberry leaf protein, polysaccharide, flavone and 1-deoxynojirimycin, which is a co-production technical scheme that mulberry leaves are used as raw materials to prepare 60% mulberry leaf protein as a first product, 60% mulberry leaf polysaccharide as a second product, 80% mulberry leaf flavone as a third product and 90% 1-deoxynojirimycin as a fourth product. The method specifically comprises the following steps: (1) primary extraction of mulberry leaves; (2) secondary extraction of mulberry leaves and separation and purification of flavone components; (3) separation and purification of the 1-deoxynojirimycin; (4) extracting mulberry leaves for the third time and separating and purifying protein; and (5) separating and purifying the mulberry leaf polysaccharide. Four products are obtained from the raw material mulberry leaves through continuous extraction and separation, the cost of a single product is reduced, the production efficiency is improved, the solvent dosage is reduced, the energy consumption is reduced, and benefits are maximized; the process operation is convenient, rapid and effective, and the comprehensive utilization degree of mulberry leaf resources is improved. The method is suitable for mulberry leaf resource development and utilization.

The present invention belongs to the field of industrial biotechnology, and discloses a method for improving the ability of Bacillus amyloliquefaciens to synthesize 1-deoxynojirimycin, the ybaS gene and its application. The method includes introducing the ybaS gene, an expression cassette containing the ybaS gene or a vector containing the ybaS gene into Bacillus amyloliquefaciens. The present invention for the first time identifies and reveals the core gene involved in DNJ biosynthesis - the ybaS gene (the nucleotide sequence is shown in SEQ ID No.1, and the amino acid sequence is shown in SEQ ID No.2), and the protein encoded by it has unique functions, filling the key gap in the DNJ synthetic metabolic pathway, providing a new perspective for analyzing the molecular mechanism of microbial synthesis of DNJ, and laying a solid molecular foundation for subsequent genetic engineering modification.

The invention discloses a method for extracting 1-deoxynojirimycin from fresh mulberry leaves, and relates to the technical field of pharmacological active ingredient extraction. The method comprises the following steps: S1, collecting fresh mulberry leaves, airing, crushing and sterilizing; s2, water is used as a solvent, continuous countercurrent extraction is adopted, and a pre-extracting solution is prepared; s3, removing impurities from the pre-extracting solution by using a micro-filtration membrane to prepare a micro-filtration permeate liquid; s4, pressurizing the microfiltration permeate liquid to permeate through a large-aperture nanofiltration membrane to prepare a nanofiltration permeate liquid; s5, pressurizing the nanofiltration permeate liquid, and concentrating the nanofiltration permeate liquid through a small-aperture nanofiltration membrane to prepare a concentrated extracting solution; and S6, performing multi-effect concentration on the concentrated extracting solution, and performing spray drying to obtain a finished product. The method is beneficial to resource conservation, energy conservation and environmental protection, and is suitable for industrial production.

The application provides an amphiphilic nobeliatin molecule, a preparation method thereof and a blood glucose lowering and antibacterial application to a diabetic wound. The application relates to the technical field of biomedical materials. The application synthesizes a novel amphiphilic nobeliatin molecule by taking 1-deoxynobeliatin as a hydrophilic part and an alkyl chain as a hydrophobic part. The compound can self-assemble into a cluster sugar molecule in an aqueous solution, has good blood glucose lowering activity through the "multivalent effect" of the cluster sugar-glycosidase, has good antibacterial activity through the tertiary amine structure of the azasugar molecule and the long alkyl chain and the bacterial membrane, and realizes the compatibility of the blood glucose lowering effect and the antibacterial effect. The synthesis raw material of the nobeliatin molecule provided by the application is rich in source, low in price, simple in preparation process and convenient for industrial application. Therefore, the discovery of the compound is expected to provide a promising treatment scheme for the development of blood glucose lowering and antibacterial integrated materials and the treatment of diabetic wounds, and has a wide potential application field.

The invention provides a bacillus amyloliquefaciens fermentation medium for producing 1-deoxynojirimycin at high yield, a fermentation method and application. The bacillus amyloliquefaciens fermentation medium comprises a carbon source, a nitrogen source and inorganic salt, the carbon source is starch, and the nitrogen source is peptone. A nitrogen source and a carbon source in a bacillus amyloliquefaciens fermentation culture medium are optimized, starch is used as the carbon source, peptone is used as the nitrogen source, and inorganic salt can balance osmotic pressure in the culture medium, maintain stability of an intracellular environment and ensure normal physiological functions and morphological structures of bacillus amyloliquefaciens cells; the bacillus amyloliquefaciens strain has the advantages that the bacillus amyloliquefaciens strain is high in yield of 1-deoxynojirimycin, energy metabolism and substance synthesis of the bacillus amyloliquefaciens are promoted, so that the yield of 1-deoxynojirimycin is increased to 1102 mg / L, the yield of 1-deoxynojirimycin is greatly increased, the production cost is reduced, and the bacillus amyloliquefaciens strain has important guiding significance on large-scale fermentation of the bacillus amyloliquefaciens.

A method for simultaneously preparing a chlorophyll paste and 1-deoxynojirimycin includes the following steps: (1) drying a silkworm excrement in a shaded area until the silkworm excrement reaches a specific moisture content; (2) placing the silkworm excrement into an extraction tank, adding an extraction solvent to obtain a mixture, and stirring the mixture with a water bath, filtering the mixture, the extraction solvent comprising both a non-polar solvent and a polar solvent; (3) subjecting the mixture to a coarse filtration to obtain a crude filtrate; (4) subjecting the crude filtrate to centrifugation, obtaining a chlorophyll-rich light phase and a 1-deoxynojirimycin-rich heavy phase; and (5) undergoing fine filtration and concentration with the chlorophyll-rich light phase to obtain a chlorophyll paste, and subjecting the 1-deoxynojirimycin-rich heavy phase with fine filtration, concentration, formulation, and spray drying to prepare 1-deoxynojirimycin.

The invention provides an amphipathic nojirimycin molecule, a preparation method thereof and application of the amphipathic nojirimycin molecule in reducing blood glucose and resisting bacteria on diabetic wounds. The invention relates to the technical field of biomedical materials. According to the invention, 1-deoxynojirimycin is taken as a hydrophilic part, an alkyl chain is taken as a hydrophobic part, and a novel amphiphilic nojirimycin molecule is synthesized. The compound can be self-assembled into a cluster sugar molecule in an aqueous solution, and has good hypoglycemic activity through a'multivalent effect 'of cluster sugar-glycosidase; through the action of a tertiary amine structure of an azasugar molecule and a long alkyl chain with a bacterial membrane, the azasugar has good antibacterial activity, and compatibility of a hypoglycemic effect and an antibacterial effect is realized. The nojirimycin molecule provided by the invention is rich in synthesis raw material source, low in price, simple in preparation process and convenient for industrial application. Therefore, the discovery of the compound is expected to provide a promising treatment scheme for development of blood glucose reducing and antibacterial integrated materials and treatment of diabetic wounds, and the compound has a wide potential application field.

To provide treatment of glycogen storage disease (GSD).SOLUTION: The present invention relates to a kit of parts comprising (i) a pharmacological chaperone or a pharmaceutically acceptable salt thereof and (ii) a therapeutic acid alpha glucosidase (GAA) polypeptide or a nucleic acid molecule encoding a therapeutic GAA polypeptide, wherein the pharmacological chaperone is 1-deoxynojirimycin (DNJ) or a derivative thereof and Ambroxol (ABX) or a derivative thereof.SELECTED DRAWING: None

The invention discloses a method for preparing high-purity 1-deoxynojirimycin, and relates to the technical field of alkaloid extraction and separation, and the 1-deoxynojirimycin is prepared from mulberry leaf powder obtained by crushing mulberry leaves through compound enzyme enzymolysis, ethanol aqueous solution extraction, nanofiltration membrane purification, resin adsorption, impurity removal and elution, concentration and gradient crystallization. According to the method, the extraction rate of the 1-deoxynojirimycin is increased by using the compound enzyme, and the purification difficulty is reduced by combining the steps of nanofiltration membrane purification, gradient crystallization and the like, so that the high-purity 1-deoxynojirimycin is obtained.

Disclosed are a 1-deoxynojirimycin derivative and use thereof. According to the present invention, a variety of 1-deoxynojirimycin derivatives are prepared and obtained through screening. Compared with 1-deoxynojirimycin as a lead compound, the 1-deoxynojirimycin derivative can better bind to an amino acid site that is related to a target protein OPA1, stabilize a binding pocket at a dimer interface, promote the formation of an OPA1 dimer and repair a mitochondrial ultrastructure, thereby significantly saving mitochondrial functions and effectively improving a physiological state of cells. The 1-deoxynojirimycin derivative of the present invention can be used for preparing a drug for treating a disease that is related to unbalanced formation of the OPA1 dimer, and for example, can be used as a potential therapeutic drug for mitochondrial cardiomyopathy and other mitochondrial diseases.

The invention belongs to the technical field of industrial biology, and discloses bacillus amyloliquefaciens capable of producing 1-deoxynojirimycin and application of the bacillus amyloliquefaciens to fermentation production of the 1-deoxynojirimycin. The yield of DNJ produced by fermentation of the bacillus amyloliquefaciens is high, the titer of the DNJ reaches 1.4 g / L or above, little foam is generated in the fermentation process, process control of industrial-grade large-scale fermentation is facilitated, the fermentation period is short, a culture medium formula is simple, and industrial production is facilitated.

The invention belongs to the technical field of industrial biology, and discloses a method for improving a 1-deoxynojirimycin biological production strain, a ybbA gene and application, and the method comprises the step of introducing the ybbA gene, an expression cassette containing the ybbA gene or a carrier containing the ybbA gene into bacillus amyloliquefaciens. According to the invention, the ybbA gene in bacillus amyloliquefaciens is subjected to comprehensive functional analysis by using an accurate gene knockout and back-filling technology. The invention aims to clarify the exact effect of the ybbA gene in DNJ biosynthesis, and provides scientific basis and theoretical support for developing a novel and efficient DNJ production strategy. The research achievement of the invention is expected to break through the bottleneck of DNJ yield increase, and opens up a new path and method for related research in the field of industrial biotechnology.

The present invention belongs to the field of plant cultivation technology and discloses a cultivation method for increasing 1-deoxynojirimycin in mulberry leaves. The present invention mixes base fertilizer into the cultivation medium and plants mulberry trees, applies Bacillus subtilis in the form of root irrigation during the mulberry growth period, and harvests mulberry leaves in a timely manner. The cultivation method of the present invention can promote the accumulation of DNJ in mulberry leaves and can reduce the level of nitrate nitrogen in the soil.

The present invention discloses 1-deoxynojirimycin derivatives and their uses. Through screening, several 1-deoxynojirimycin derivatives were prepared. Compared with the lead compound 1-deoxynojirimycin, these derivatives better bind to the relevant amino acid site of the target protein OPA1 and are stabilized in the binding pocket of the dimer interface. They promote the formation of OPA1 dimers and restore mitochondrial ultrastructure, thereby significantly maintaining mitochondrial function and effectively improving cellular physiological status. The 1-deoxynojirimycin derivatives of the present invention are useful for preparing drugs for treating diseases associated with an imbalance in OPA1 dimer formation, and are promising therapeutic agents for, for example, mitochondrial cardiomyopathy and other mitochondrial diseases.

The invention provides a deoxynojirimycin-pterostilbene compound as well as a preparation method and application thereof, and relates to the technical field of medicinal chemistry. The deoxynojirimycin-pterostilbene compound provided by the invention has a structure as shown in a formula I, and n is the number of carbon and is selected from 2-6, 8, 10 or 12. The invention also provides the application of the deoxynojirimycin-pterostilbene compound in preparation of medicines for treating or preventing abnormal glucose and lipid metabolism diseases. According to the prepared deoxynojirimycin-pterostilbene compound, the adopted method has the advantages that raw materials are easy to obtain, operation is easy and convenient, and the yield is high, a target compound can be rapidly synthesized, and the target compound has the effect of reducing blood sugar and blood lipid.

The composition is prepared from folium mori, radix astragali, radix puerariae, rhizoma dioscoreae, poria cocos and pericarpium citri reticulatae in specific parts by weight, and the core of the composition is that the contents of key active ingredients 1-deoxynojirimycin and total flavonoids in the folium mori raw material are quantitatively screened; the preparation method adopts a step-by-step water extraction technology and a low-temperature concentration technology. The invention relates to the technical field of health food and traditional Chinese medicine pharmacy. By performing standardized quality control on the core raw materials and combining scientific compound compatibility and an optimized preparation process, the problems that in the prior art, due to the fact that the quality of the raw materials is not uniform, the product effect is unstable, and due to the simple formula, the action target spot is single are solved, and the prepared composition has the stable and efficient auxiliary blood sugar reducing effect and is suitable for being used for preparing the traditional Chinese medicine composition. And the composition can synergistically improve the metabolic state of the body, is high in safety and has remarkable popularization and application values.

The present invention provides a dual-targeted polyhydroxy alkaloid compound, a preparation method and uses thereof, relating to the field of medicine. The dual-targeted polyhydroxy alkaloid compound provided by the present invention combines 1-deoxynojirimycin with thiazolidinedione to synthesize a novel 1-deoxynojirimycin derivative having effects such as treating diabetes. The synthesis method is simple and the product purity is high. The derivative has excellent α-glucosidase inhibitory activity and can directly bind to PPARγ, thereby improving insulin resistance and enhancing the blood glucose lowering effect.

Use of a mulberry extract in the preparation of a medicament for treating and / or preventing diabetic vascular complications. The mulberry extract comprises an alkaloid, and the alkaloid comprises 1-deoxynojirimycin. The diabetic vascular complications include diabetic foot or diabetic atherosclerosis. The mulberry extract has a significant ameliorating effect on the diabetic vascular complications.

This invention discloses a high-yield 1-deoxynojirimycin-producing *Bacillus pumilus* strain and its applications, belonging to the field of microbiology. This invention, through screening, ARTP and NTG mutagenesis, obtained a *Bacillus pumilus* strain with significantly increased 1-deoxynojirimycin production. Bacillus pumilus DNJ-BP01, deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 38218, was used to inoculate the seed culture of Bacillus pumilus DNJ-BP01 into a fermentation medium containing 5% beef extract supplemented with 2.5% mannitol as the carbon source and 3% potassium nitrate as the nitrogen source. After 76 hours of aerobic fermentation, the yield of 1-deoxynojirimycin reached 0.892 g / L. This invention provides a new fermentation strain for the efficient industrial production of 1-deoxynojirimycin and has broad application prospects.

The invention belongs to the technical field of biological medicines, and particularly relates to a method for preparing a synergistic hypoglycemic active composition based on specific symbiotic fermentation and application of the synergistic hypoglycemic active composition. The invention discloses a method for preparing a synergistic hypoglycemic active composition based on specific symbiotic fermentation and application thereof, and aims to solve the problems of complex process, high cost and incapability of synergistic interaction of active components caused by physical mixing or respective treatment of mulberry leaves and white kidney beans. The method comprises the following steps: mixing mulberry leaf powder and white kidney bean powder according to a ratio of 1: (0.5-2), inoculating a compound lactic acid bacteria agent compounded by lactobacillus plantarum and lactobacillus mucus, carrying out solid symbiotic fermentation at 30-37 DEG C for 60-84 hours, and carrying out extraction, centrifugation, concentration and freeze drying to obtain the composition. The obtained product is rich in 1-deoxynojirimycin, flavonoid aglycones, small molecule peptides and novel double-enzyme inhibition metabolites, has remarkable alpha-amylase and alpha-glucosidase synergistic inhibition activity, and can be used for preparing food, health food or medicine with a blood glucose reducing function.

The present application provides novel iminosugars and their use as glucosidase inhibitors. The present inventors have discovered that certain deoxynojirimycin derivatives may be effective in inhibiting glucosidases. In particular, such deoxynojirimycin derivatives may be useful for treating a disease or condition where inhibiting glucosidase may be important.

The present invention relates to the technical field of extraction of mulberry leaves after frost, and in particular to an extraction method and application of an extract of mulberry leaves after frost. The extraction method sequentially includes pretreatment of mulberry leaves after frost, preparation of a composite enzyme aqueous solution, preparation of a hyaluronic acid aqueous solution, enzymatic hydrolysis treatment, and extraction of an extract of mulberry leaves after frost. Through specific selection of raw materials and cooperation with a specific extraction process, an extract of mulberry leaves after frost with a 1-deoxynojirimycin mass content as high as 5.2-7.0% is obtained; at the same time, it breaks through the limitation that the existing mulberry leaf extract needs to be stored under harsh conditions of 2-8°C. After the extract of mulberry leaves after frost extracted by a specific extraction process is stored in the dark at 18°C for 300 days, the retention rate of the mass content of 1-deoxynojirimycin it is rich in can be greater than or equal to 89%, further reducing the storage cost for its subsequent popularization as a hypoglycemic product.

The application discloses a method for detecting deoxy-najinmycin in milk and dairy products. The method comprises the following steps: extracting a sample solution to be derivatized by using an extracting agent, adding a buffer solution and a derivatization reagent to the sample solution to be derivatized to perform derivatization, performing liquid chromatography test on the sample to be tested after the derivatization, and qualitatively and quantitatively determining the sample according to the retention time and external standard method. The content of deoxy-najinmycin in the sample to be tested is obtained according to the test result. The extracting agent is an acetonitrile aqueous solution, the buffer solution is a potassium borate buffer solution, and the derivatization reagent comprises a fluorenylmethyloxycarbonyl chloride acetonitrile solution, a glycine solution and an acetic acid aqueous solution. The precision, repeatability, recovery rate and standard curve linearity of the detection method meet the industry requirements. The method has good extraction effect, high sensitivity, stable and accurate results, and is simple to operate and convenient to popularize.

The invention relates to the technical field of genetic engineering and microbiology, and discloses bacillus amyloliquefaciens for highly producing 1-deoxynojirimycin as well as a construction method and application of the bacillus amyloliquefaciens. The yield of 1-deoxynojirimycin (1-DNJ) in bacillus amyloliquefaciens is increased by knocking out the secondary thiamine phosphate synthase gene yjbQ. According to the bacillus amyloliquefaciens engineering bacterium LX-12 yjbQ, on the basis of plasmid T2 (2)-Ori, a knockout vector T2-delta yjbQ of a secondary thiamine phosphate synthase gene yjbQ is constructed, and the gene yjbQ is successfully knocked out from bacillus amyloliquefaciens LX-12, so that the bacillus amyloliquefaciens engineering bacterium LX-12 yjbQ is obtained. Compared with the bacillus amyloliquefaciens LX-12, the yield of the 1-deoxynojirimycin J of the engineering strain LX-12 yjbQ is at least increased by 26.17%, and the highest yield can reach 37.25%.