39 results about "Galactosidases" patented technology

This invention provides a method for screening β-galactosidase inhibitors, belonging to the field of pharmaceutical technology. The first step of this method uses fluorescence sensing technology to rapidly identify natural products with β-Gal inhibitory activity. The principle is based on the fluorescence quenching property of p-nitrophenol (PNP) on sulfur quantum dots (SQDs); β-Gal catalyzes the hydrolysis of p-nitrophenyl-β-D-galactopyranoside (PNPG) to generate the specific product PNP; in the presence of β-Gal, PNPG can be enzymatically catalyzed to generate PNP, quenching the fluorescence of SQDs; when screened for β-Gal inhibitory activity, the fluorescence of SQDs can be restored. The second step uses affinity chromatography to capture components in samples with β-Gal inhibitory activity. This method involves immobilizing β-Gal on a support material, eluting with a solvent to obtain components that can bind to β-Gal, and then using liquid chromatography-mass spectrometry (LC-MS) to identify the components. This method can rapidly and accurately screen β-Gal inhibitors from natural products.

The application discloses a lactobacillus crispatus based on multiple targets to realize anti-aging effect and application of transformation of glycyrrhiza for efficiency enhancement, and belongs to the technical field of microorganisms and the technical field of medicine. The lactobacillus crispatus CCFM1362 provided in the application can ferment glycyrrhiza extract, can improve the content of glycyrrhizic acid, hemiglycyrrhizin isoflavone B, naringenin chalcone and other substances, can enhance the efficacy of glycyrrhiza extract, and can realize the effect of multiple targets for anti-aging, and the specific embodiments are as follows: (1) the content of beta-galactosidase in liver tissue is inhibited; (2) the level of aging-related secretory phenotype in liver tissue is reduced; (3) the Nrf2 related pathway is activated, and the level of oxidative stress is reduced; (4) the NF-kappa B related pathway is activated, and the level of chronic inflammation is reduced. Therefore, the lactobacillus crispatus CCFM1362 and the glycyrrhiza extract have great application prospect in the production of anti-aging products.

Levels of expression of antibiotic resistance genes are increased up to six-fold by inserting a proteasome-targeting tag into transgenes expressed in eukaryotic cells. Various selectable marker proteins are combined with different destabilization domains, leading to up to 70% increase in transgene expression. The increase in expression varies highly depending on the engineered construct and the lines cells used. Increase in expression drives exosome loading of cargo proteins in some aspects. By increasing expression and by editing trafficking signals of cargo proteins, proteins that normally locate to the ER can be trafficked to exosomes. This disclosure discloses efficient exosome delivery of a wide variety of engineered proteins, including modified antigen proteins of SARS-CoV-2 and influenza, and other proteins such as a modified alpha galactosidase A, an extracellular domain of vascular endothelial growth factor fused to a constant region of a human immunoglobulin heavy chain, and modified trastuzumab heavy and light chains.

The application discloses a beta-galactoside enzyme mutant and application thereof. The application takes wild-type beta-galactoside enzyme Bgal1-3 as a parent to provide a mutant with improved thermal stability, enzyme activity and / or oligogalactose production capacity compared with the parent. Meanwhile, the application takes the mutant M9 of Bgal1-3 as a parent to provide a mutant with improved hydrolysis activity compared with the parent. The beta-galactoside enzyme mutant can be used for converting lactose to produce low / zero-lactose or oligogalactose-rich dairy products, and is beneficial to the development of low / zero-lactose or oligogalactose-rich dairy product related industry.

Disclosed are use of a circRNA molecule as a diagnostic marker in a cell senescence diagnosis kit, and use thereof as a treatment target for cell senescence. The present invention finds that circHERC1 is lowly expressed in both cell models of natural senescence and induced senescence. Overexpression of circHERC1 in endothelial cells can improve the proliferation capacity of the endothelial cells, effectively extend the telomere length of the cells, activate telomerase, reduce the β-galactosidase positive rate of the cells, and reduce the expression of all senescence-associated proteins. Introduction of exogenous circHERC1 into mice can effectively delay the senescence of various organs and the shortening of telomeres in aged mice and reduce the β-galactosidase positive rates of various organs of aged mice.

This invention discloses *Bifidobacterium longum* with anti-aging functions and its applications, belonging to the field of microbial technology. The *Bifidobacterium longum* BBS 02 provided by this invention can promote cell vitality and proliferation, inhibit the high activity of β-galactosidase in senescent cells, and inhibit the high expression of the cyclin-dependent kinase inhibitor gene Cdkn1a, thereby achieving an anti-aging effect. The *Bifidobacterium longum* BBS 02 can be used to prepare edible nutrients or skin nutrients.

The invention relates to a lyophilized preparation containing alpha-galactosidase A fusion protein and a preparation method of the lyophilized preparation.

The present application relates to the use of a substance that regulates BCL11A gene and / or its encoded product in the preparation of a drug for reversing skin photoaging, wherein the skin photoaging includes skin cell aging caused by sunlight, light, etc., and the skin cells include keratinocytes, dermal fibroblasts, melanocytes. The regulation is to up-regulate the expression of BCL11A gene and / or promote the production of its encoded product. Typical clinical manifestations of the skin photoaging include wrinkles, dry skin, loose skin, dilated capillaries, uneven skin pigmentation, and skin itching. The substance that regulates BCL11A gene and / or its encoded product includes a substance that regulates BCL11A gene DNA molecules and a substance that regulates the expression of BCL11A translation product protein molecules. The reversal of skin photoaging is manifested in reducing the proportion of cells positive for aging-related beta-galactosidase staining in skin cells and reducing the expression of aging-related protein p16 and DNA damage protein gammaH2AX.

This invention relates to a recombinant Bacillus subtilis strain producing high-activity β-galactosidase and its application. The recombinant Bacillus subtilis strain uses Bacillus subtilis WG600 as the host strain, derived from... Enterobacter Using sp. β-galactosidase as the target gene, through the promoter (P) phrE Synergistic optimization of the NCS sequence (glnA-30bp, MLD42) and 5'-UTR (UTR13, UTR4, UTR7) enabled the fermentation enzyme activity of Ensp-β-galactosidase in Bacillus subtilis to reach over 500 U / mL, with a maximum of 584.2 U / mL. Compared with existing technologies, the enzyme activity has been greatly improved, representing the highest level of enzyme activity for β-galactosidase production in Bacillus subtilis, which can meet the demand for high-activity enzymes in industrial production.

This invention discloses a method for preparing a water-soluble homogeneous heteropolysaccharide from highland barley and its novel application in promoting the expression of β-galactosidase in the small intestinal mucosa, belonging to the field of polysaccharide preparation and application technology. This invention provides a method for preparing the water-soluble homogeneous heteropolysaccharide HBP-1a from highland barley, involving extraction, alcohol precipitation, decolorization, and deproteinization of highland barley raw materials, followed by separation and purification using anion exchange column chromatography and gel column chromatography. The water-soluble homogeneous heteropolysaccharide HBP-1a was successfully isolated from the highland barley polysaccharide. This invention is the first to discover that the water-soluble homogeneous heteropolysaccharide HBP-1a can effectively promote the expression of β-galactosidase in the small intestinal mucosa and can be used to improve symptoms related to lactose intolerance diarrhea. This invention explores a new use for the water-soluble homogeneous heteropolysaccharide HBP-1a from highland barley, broadening its application fields and possessing broad prospects and market value.

Provided is a method for preparing lactulose using immobilized β-galactosidase. The method comprises the following steps: pretreating a macroporous weakly basic styrene-based anion exchange resin; mixing the pretreated resin with a β-galactosidase solution and adding same to a buffer, adsorbing same using a shaker, and leaving same to stand after adsorption; then adding a glutaraldehyde solution for cross-linking, and discarding the supernatant to obtain an immobilized enzyme column; adding a magnesium ion solution to a mixed solution of lactose and fructose, and then converting same by means of the immobilized enzyme column to obtain a lactulose conversion solution; and sequentially performing decolorization, ion exchange, vacuum concentration, and sequential simulated moving bed chromatography separation on the lactulose conversion solution to obtain a lactulose solution.

A polynucleotide is provided comprising a nucleotide sequence encoding a biomolecule (e.g., human α-galactosidase A), a nucleotide sequence encoding a promoter (e.g., a hepatocyte-specific promoter or a hepatocyte-myocyte dual-specific promoter), or a nucleotide sequence encoding an expression cassette (e.g., a human α-galactosidase A expression cassette). Furthermore, promoters, expression cassettes, vectors, host cells, gene delivery systems (e.g., recombinant adeno-associated virus (AAV) particles and recombinant viral particles such as nonviral gene delivery systems), related pharmaceutical compositions, and methods for using them are also provided. Such compositions and methods are particularly suitable for gene therapy, and especially for lysosomal storage disorders, including Fabry disease.

The invention discloses an application of a 2-aminothiazole derivative in preparation of an anti-aging drug. The 2-aminothiazole derivative can effectively relieve senescence of microglial cells in vitro, reduce accumulation of senescence-related beta-galactosidase in senescence cells, reduce secretion of senescence-related secretion phenotypes and improve senescence cell apoptosis and oxidative stress loss. In a mouse in-vivo experiment, the compounds A1-A10 can remarkably improve cognitive impairment and long-term and short-term learning and memory ability damage caused by aging, reduce the transcription expression level of proinflammatory factors in the brain and relieve neuroinflammation. In conclusion, the 2-aminothiazole derivative provided by the inventor has a good anti-aging curative effect and has huge application potential in the anti-aging aspect.

The present disclosure provides less immunogenic mutant α-galactosidase A protein (α-gal), methods of making, and methods of use. The less immunogenic mutant α-gal of the present invention provides a reduced immune response when administered to a subject. Methods of treating Fabry in a subject are also provided, the methods comprising administering the disclosed less immunogenic mutant α-gal.

Disclosed in the present invention is a use of a 2-aminothiazole derivative in the preparation of an anti-aging drug. The 2-aminothiazole derivative can effectively alleviate microglial cell aging in vitro, reduce accumulation of senescence-associated β-galactosidase in aged cells, decrease secretion of a senescence-associated secretory phenotype, and ameliorate apoptosis and oxidative stress damage in the aged cells. In a mouse in vivo experiment, compounds A1-A10 can significantly improve aging-induced cognitive dysfunction and impairment of long-term and short-term learning and memory abilities, reduce transcriptional expression levels of pro-inflammatory factors in the brain, and alleviate neuroinflammation. In summary, the present inventors propose that 2-aminothiazole derivatives have good anti-aging efficacy and possess great application potential in anti-aging.

The application discloses an engineered bacillus subtilis strain for producing tagatose, a construction method and application thereof, and belongs to the technical field of bioengineering. The engineered strain takes bacillus subtilis as a host, co-expresses L-arabinose isomerase from Erysipelothrix larvae and beta-galactosidase from escherichia coli, and constructs the bacillus subtilis for co-expressing double enzymes through ribosome binding sites or promoters in series. The double-enzyme bacillus subtilis whole cell catalyzes lactose to produce D-tagatose, and the yield and conversion efficiency of D-tagatose are significantly improved after optimization of the enzymatic reaction. The application adopts single-strain double-enzyme co-expression and one-step whole-cell catalysis, uses low-cost lactose as a substrate, and has the advantages of mild reaction, less by-product, simplified process, no endotoxin safety hazard, and the like. The application solves the problems of low conversion rate, high substrate cost and complicated strain compounding in the prior art, and is suitable for industrialized production of food-grade D-tagatose.

The application discloses a kind of umbilical cord stem cell quality analysis method and system using image processing, it is related to cell quality analysis technical field, the method includes: the first microscopic image of the umbilical cord stem cell of cultivation is collected, the stem cell morphology identification is carried out to first microscopic image, obtain multiple stem cell morphology information, quality identification is carried out, and obtain first quality information;Carry out beta-galactosidase dyeing image processing collection, obtain second microscopic image, and obtain multiple blue morphological information and second quality information by identification;The similarity of multiple blue morphological information and multiple stem cell morphology information is analyzed, and first confidence is obtained, and the precipitation crystallization analysis of multiple blue morphological information is carried out, and second confidence is obtained by processing;According to first confidence and second confidence, in combination with the first quality information and second quality information are calculated, and quality information is obtained.The application solves the technical problem that stem cell quality analysis is not accurate in the prior art.

FIELD: neurology; laboratory diagnostics.SUBSTANCE: used for the differential diagnosis of Parkinson's disease not associated with mutations in the GBA1 gene, and multiple system atrophy. The patient's peripheral venous blood is collected, from which mononuclear cells are isolated by gradient centrifugation, followed by their differentiation into a primary culture of macrophages in the presence of the macrophage colony-stimulating growth factor M-CSF to ensure proliferation and differentiation of monocytes into mature macrophages. The obtained blood macrophage cells are applied to 903 filter cards at a concentration of 2×106 cells / ml. Whatman 903 Sample Collection Cards can be used as 903 filter cards. The activity of lysosomal enzymes is determined: glucocerebrosidase GCase, alpha-galactosidase GLA, acid sphingomyelinase ASMase, galactosylceramidase GALC by high-performance liquid chromatography in combination with tandem mass spectrometry. The value of the canonical linear discriminant function CLDF is calculated using the stated formula. If the value of CLDF is ≥ 43.44, the patient is diagnosed with multiple system atrophy. If the value of CLDF is < 43.44, Parkinson's disease is diagnosed in patients who do not have a mutation in the GBA1 gene.EFFECT: method enables reliable and accurate differential diagnosis of Parkinson's disease and multiple system atrophy by assessing the activity of lysosomal enzymes.2 cl, 2 dwg, 2 ex

The application belongs to the field of biological detection, and particularly relates to a method for detecting nucleosome histone and its modifier based on single molecule immunological technology. Based on single molecule technology, the nucleosome sample to be detected which is tailed by polyadenylic acid (Poly-A) is specifically captured by antibody modified magnetic beads, then hybridized with biotin labeled polythymidylic acid (Poly-T) to form an immunocomplex, and then recognized by streptavidin labeled beta-galactosidase (SbetaG) in a single molecule array chip, so that the visualization, numerical quantification and detection of the sample circulating nucleosome and its modifier are realized. The method has high sensitivity, repeatability and wide dynamic detection range, and can realize the accurate quantification of histone H3 and its six key epigenetic modifiers in circulating nucleosome by using only 20 muL of serum sample, which is beneficial to reveal the variation of circulating nucleosome histone modifier in the early development stage of various diseases, and has good clinical application prospect.

This invention relates to the field of biotechnology, specifically to a high-yield α-galactosidase-producing mutant strain of *Weizmannii coagulans* and its application. This invention is the first to propose using *Weizmannii coagulans* as an α-galactosidase-producing bacterium. Through mutagenesis, a high-yield α-galactosidase-producing mutant strain was obtained. The fermentation broth of the mutant strain showed a 4.37-fold increase in α-galactosidase activity compared to the original strain, maintaining over 60% activity within a pH range of 6.0–10.0, and exhibiting 1.80 times better heat resistance than the original enzyme. Furthermore, it can hydrolyze the anti-nutritional factors mesenteryne, raffinose, and stachyose in soybean meal, demonstrating high industrial value in the fermented soybean meal and feed processing industries. This invention lays the foundation for the application of *Weizmannii coagulans* in the manufacturing and processing of α-galactosidase.

Disclosed in the present invention are a Pichia pastoris genetically engineered strain for the efficient surface display of an enzyme based on an adhesion protein, and a construction method therefor and the use thereof in immobilized continuous catalysis. An adhesion protein gene, a β-galactosidase gene LacA, and a cell wall anchoring protein gene Pir1p are expressed in the starting strain, Pichia pastoris. The Pichia pastoris genetically engineered strain is obtained by integrating and expressing a cell wall adhesion protein gene into the genome of Pichia pastoris GS115, integrating a β-galactosidase surface display gene in multiple copies, and displaying a target protein, β-galactosidase, on a yeast surface under the control of a constitutive promoter. The Pichia pastoris genetically engineered strain is capable of performing multi-batch continuous catalysis, and exhibits a catalytic effect superior to that of the enzyme surface display genetically engineered strain without an integrated biofilm gene.

The application discloses application of a composite biological enzyme preparation in preparation of a product for improving chemical liver injury, and relates to the technical field of biotechnology.The raw materials of the composite biological enzyme preparation include 3-14 kinds of papain, bromelain, acid protease, pepsin, trypsin, alpha-amylase, sucrase, lipase, cellulase, pectinase, glucoamylase, nuclease, serrapeptase and beta-galactosidase.The composite biological enzyme preparation can significantly improve chemical liver injury, effectively reduce serum ALT and AST activity, relieve liver cell degeneration, necrosis and inflammatory infiltration, and reduce liver tissue inflammatory factor level;for alcoholic liver injury, the composite biological enzyme preparation can improve serum and liver ADH and ALDH activity, promote alcohol metabolism, regulate four blood lipid indexes, reduce liver index and liver MDA content, and reduce liver oxidation damage.

The present application relates to the technical field of fluorescent probe, and discloses a fluorescent probe locked by viscosity and glycosidase, and a preparation method and application thereof.The present application provides a compound, the fluorescence emission intensity of which is enhanced with the enhancement of the solution viscosity; meanwhile, after incubation with beta-galactosidase, the fluorescence intensity of the compound at 640nm is enhanced.Therefore, in the early diagnosis imaging of ovarian cancer, the compound can be used as a fluorescent probe locked by viscosity and glycosidase, and is used for accurate imaging of ovarian cancer.The fluorescent probe of the present application greatly improves the selectivity of the probe by using a double activation mechanism, effectively avoids non-specific activation to normal tissues, and is particularly suitable for targeted imaging of specific pathological environments such as tumors.

The application relates to the field of biological medicine, and particularly relates to a WDR6 gene and application thereof Wdr6 in preparation of a liver aging delaying product. Wdr6 Low expression can aggravate the liver aging degree, and liver-specific Wdr6 Overexpression can significantly down-regulate the expression of a p21 protein, which is a marker of aging, in liver tissues of naturally aging mice, and the number of cells that are positively colored by aging-related beta-galactosidase staining is significantly reduced, indicating that Wdr6 Overexpression can effectively maintain liver young homeostasis and delay liver aging. Meanwhile, through an in-vitro culture of a liver cell line aging model induced by bleomycin, it is verified that the WDR6 protein can increase the expression of a long-life protein SIRT1 protein, down-regulate the expression of a p21 protein, and reduce beta-galactosidase, so as to realize liver aging delay.

This invention discloses an anti-aging compound, its preparation method, and its applications. Experimental results in in vitro anti-aging screening models using MEF cells and MRC-5 cells show that the compound significantly reduces the proportion of cells positive for aging-related β-galactosidase staining, downregulates the protein expression levels of aging markers p53, p21, and p16, and restores the proliferative capacity of senescent cells. In *C. elegans*, the compound prolongs the lifespan of the nematodes, improves their motility, and reduces lipofuscin accumulation due to aging. Therefore, this compound can be used in the preparation of anti-aging drugs and health products. This invention provides new candidate compounds and experimental evidence for the development of new anti-aging drugs and basic research, and has significant practical implications.

The present application relates to compounds of formula (I) and salts and solvates thereof, wherein A1, A2, A3 and R1, R2, R 3a , R 3b , R4 and R5 are as defined in the description, and their use for the treatment and / or prevention of disorders and conditions associated with an altered activity of β-galactosidase, in particular galactoside β-1 or GLB1, including GM1 gangliosidosis and Morquio syndrome B.

The application belongs to the technical field of enzyme activity and active oxygen content detection, and particularly relates to a fluorescent probe for tracing beta-galactosidase, lysosome pH and active oxygen in the aging process. The application discloses a fluorescent probe capable of efficiently and selectively recognizing beta-galactosidase, lysosome pH and active oxygen, and having high sensitivity. Based on the spectral and imaging research at the cell and biological levels, it is shown that the fluorescent probe can track the aging process, has good stability in terms of sensitivity, anti-interference ability and imaging, and has good market application value for early diagnosis of aging.

The application belongs to the technical field of biological analysis detection, and specifically discloses a kind of beta-galactosidase fluorescent probe and a preparation method and application thereof.The molecular structural formula of the fluorescent probe provided by the application includes one of TCFM-Gal1, TCFM-Gal2, TCFM-Gal3 or TCFM-Gal4.By introducing an eliminable fluorine-substituted methyl (difluoromethyl, monofluoromethyl) at the ortho position of the enzymatic substrate of the fluorescent probe, the generation of an electrophilic quinone methyl intermediate under enzymatic hydrolysis can be realized;and further covalently combined with the adjacent nucleophilic protein, the specific opening and in-situ retention of the fluorescent signal in the senescent cells are realized.The fluorescent probe also has excellent beta-galactosidase detection sensitivity and specificity, and the detection fluorescence has a good linear relationship with the enzyme concentration, and the detection range is wide.

The application belongs to the technical field of medicine, and particularly relates to application of matricaria o in preparation of anti-aging products. The application provides, for the first time, application of matricaria o in preparation of anti-aging products. The application innovatively discovers that matricaria o (KO), an active ingredient of sophora flavescens, has an anti-oxidative stress effect, can reduce the content of beta-galactosidase, reduce the content of ROS, and increase the mitochondrial membrane potential, and can be applied to preparation of anti-oxidative and anti-aging related drugs and / or cosmetics, and has certain application prospect and social significance.