28 results about "Glucosidases" patented technology

Disclosed herein is a method for the treatment of Pompe Disease comprising administering a recombinant AAV (rAAV) vector comprising a rAVV genome comprising a heterologous nucleic acid encoding a GAA signal peptide or portion thereof, a heterologous signal peptide, and an acid alpha-glucosidase (GAA) polypeptide, or N-terminal truncation thereof, where the heterologous nucleic acid is operatively linked to a liver-specific promoter, where the nucleic acid encoding GAA polypeptide can be wild type nucleic acid sequence, or modified nucleic acid sequence, or a codon optimized nucleic acid sequence, and can optionally be modified to reduce or completely eliminate CG and CpG dinucleotides and, optionally eliminated alternative reading frames (ARF) content.

The invention discloses a method for detecting F26G enzyme activity based on an HPLC (High Performance Liquid Chromatography) technology, and belongs to the technical field of biological enzyme activity detection. According to the method, on the basis of the characteristic that furostanol saponin 26-O-beta-D-glucosidase (F26G) hydrolyzes furostanol saponin C-26 glucosyl, a detection system which takes furostanol saponin as a substrate and quantifies substrate consumption through an HPLC technology to calculate enzyme activity is established, the method has high substrate specificity, and compared with a pNPG method, the method has the advantages that the detection efficiency is high, and the detection cost is low. The method has the advantages of high sensitivity and high accuracy, can accurately eliminate the interference of other glucosidase in the system, overcomes the defects of poor specificity, low sensitivity and difficulty in accurate determination of the enzyme activity of the traditional pNPG method, provides a new method for accurate detection of the activity of the F26G enzyme in the medicinal materials, and provides technical support for quality monitoring of the medicinal materials and decoction pieces.

Provided herein are proteins, which are capable of being transported across the blood- brain barrier (BBB) and comprise an acid alpha-glucosidase (GAA) enzyme-Fc fusion polypeptide. Certain embodiments also provide methods of using such proteins to treat Pompe disease.

The present invention provides engineered acid alpha-glucosidase (GAA) polypeptides and compositions thereof. In some embodiments, the engineered GAA polypeptides have been optimized to provide increased expression, stability at neutral pH, and activity in cell lysates. The invention also provides methods for utilization of the compositions comprising the engineered GAA polypeptides for therapeutic and other purposes.

The present invention specifically relates to an enzyme combination comprising a glucosidase enzyme, a secoisolariciresinol dehydrogenase enzyme and a glucose dehydrogenase enzyme, which are capable of carrying out reactions to convert secoisolariciresinol diglucoside (SDG) into matairesinol with a high yield. The invention also relates to the uses of the above combination, and to methods for producing matairesinol.

A method for treating Pompe disease including administration of recombinant human acid α-glucosidase having optimal glycosylation with mannose-6-phosphate residues in combination with an amount of miglustat effective to maximize tissue uptake of recombinant human acid α-glucosidase while minimizing inhibition of the enzymatic activity of the recombinant human acid α-glucosidase is provided.

The present invention provides Monosporozyma servazzii. The Monosporozyma servazzii is the Monosporozyma servazzii strain AMCC 31719, which is deposited at the China Center for Type Culture Collection (CCTCC) under the accession number CCTCC NO: M 20242063. The Monosporozyma servazzii strain of the present invention is capable of utilizing multiple carbon sources during growth and is characterized by high glycerol and β-glucosidase production, and aroma production. The strain can enhance the flavor of baked products and improve the texture thereof, and is mainly applicable in the fields of food and fermented food.

Recombinant AAV (rAAV) vectors comprising a rAVV genome comprising a heterologous nucleic acid encoding a lysosomal protein, e.g., acid alpha-glucosidase (GAA) polypeptide, and optionally a signal peptide and / or optionally a targeting sequence, e.g., IGF2 targeting peptide, operatively linked to a liver-specific promoter (LSP), enabling the GAA polypeptide to be secreted from the liver and targeted to the lysosomes. Particular embodiments relate to a recombinant AAV (rAAV) vector encoding an alpha-glucosidase (GAA) polypeptide, having a liver secretory signal peptide and a IGF2 targeting peptidethat binds human cation-independent mannose-6-phosphate receptor (CI-MPR) or to the IGF2 receptor, permitting proper subcellular localization of the GAA polypeptide to lysosomes. Also encompassed are cells, and methods to treat a lysosomal disease, for example, a glycogen storage disease type II (GSD II) disease and / or Pompe Disease with the rAAV vector.

Recombinant human alpha glucosidase (rhGAA) compositions derived from CHO cells are disclosed, comprising a more optimal glycan composition consisting of rhGAA with higher amounts of N-glycans bearing mannose-6-phosphate (M6P) or bis-M6P, along with low amounts of non-phosphorylated high mannose glycans, than conventional rhGAA, along with low amounts of terminal galactose on complex oligosaccharides. Compositions comprising the rhGAA are described, as are methods of use.

The present application provides methods of treating Pompe disease such as infantile-onset Pompe disease (IOPD) using a pharmaceutical composition comprising an oligosaccharide-acid α-glucosidase (GAA) conjugate, such as avalglucosidase alfa. Also provided are formulations of the oligosaccharide-GAA conjugates.

A method of preparing a diterpenoid composition is provided. The method comprises the following steps: extracting a lysimachia capillipes material to obtain an extracted composition; and performing a series of separation steps to obtain at least one target extract of isolated diterpenoids of lysimachia capillipes. There is also provided a diterpenoid composition obtained by the method, an oral composition comprising the diterpenoid composition, a composition comprising at least one diterpenoid extract of lysimachia capillipes, the use of the composition in the preparation of a medicament for the treatment of hyperglycemia and / or hyperlipidemia in a subject, and the use of the composition in the treatment of hyperglycemia and / or hyperlipidemia in a subject. And methods of treating hyperglycemia and / or hyperlipidemia in a subject.

The application discloses an extremely thermophilic biochar composite microbial agent and application thereof in pig manure aerobic composting, and belongs to the technical field of microorganisms. The application discloses a strain of thermophilic thermus N1209, and further discloses an extremely thermophilic biochar composite microbial agent, which comprises thermophilic glucosidase geobacillus BGSC 95A1, bacillus licheniformis ZR-1, thermus N1209 and thermophilic adipose geobacillus L5. The extremely thermophilic biochar composite microbial agent has the characteristics of high temperature resistance, higher amylase, protease and lipase activity compared with single microbial agent, shows synergistic advantages, can significantly promote the composting of pig manure, has advantages in the aspects of nitrogen element reservation and CH4 and N2O emission reduction in the pig manure composting process, and provides a new method for the treatment of poultry breeding waste.

A method is described for increasing the amount of fermentable sugars in a fermentation substrate by treating with a combination of an enzyme having transglucosidase activity and an enzyme having glucoamylase activity to hydrolyze oligosaccharides and / or polysaccharides that are typically unhydrolyzable by glucoamylases alone during fermentation. The method is most effective for fermentation substrates containing small amounts of maltose and maltotriose.

The disclosure provides a thermostable enzyme-linked biosensor expression cassette comprising a nucleic acid comprising a nucleotide sequence encoding a β-glucosidase reporter. The enzyme-linked biosensor expression cassette of the disclosure comprises a nucleic acid comprising a nucleotide sequence encoding (i) a transcription factor, (ii) a promoter, (iii) a terminator, and (iv) a cloning site for a gene of interest. The disclosure further provides novel variants of β-glucosidase that function as the reporter enzyme and exhibit superior properties (e.g., without limitation, pH stability and thermal stability) compared to existing β-glucosidase, providing improved biosensor expression cassettes.

This invention discloses a multi-step sequential enzymatic hydrolysis combined with multi-stage membrane purification method for preparing kudzu oligosaccharides, relating to the field of functional food processing technology. The preparation method includes the following steps: adding kudzu powder to water to prepare a starch slurry, then liquefying it using α-amylase, followed by enzymatic hydrolysis using debranching enzymes and glycoside hydrolases to obtain an enzymatic hydrolysate; performing membrane separation on the enzymatic hydrolysate to obtain a fraction with a molecular weight higher than 1 kDa and lower than 5 kDa, followed by drying to obtain the kudzu oligosaccharides; the debranching enzymes are pullulanase and isoamylase; the glycoside hydrolases are β-glucosidase or α-transglucosidase. This preparation method can effectively increase the proportion of the DP3-15 fraction of kudzu oligosaccharides, achieving a product purity of over 91% and a total yield exceeding 22%.

A method for hydrolyzing an alpha-1,5 glucosyl-fructose bond in a levansucrose, e.g., a sucrose, is disclosed. The method comprises contacting the sucrose with an alpha-glucosidase, e.g., a transglucosidase or a glucoamylase, under suitable conditions, during which the enzyme hydrolyzes at least one alpha-1,5 glucosyl-fructose bond of the sucrose. The method is used, e.g., to reduce the amount of levansucrose in a filtrate isolated from a glucan synthesis reaction.

The invention discloses low-temperature glucose-tolerant beta-glucosidase and application thereof, and relates to the field of enzyme engineering. The beta-glucosidase is separated from Streptomyces microflavus CLSD-1 and belongs to a GH1 family, and the amino acid sequence of the beta-glucosidase is shown as SEQ ID No: 1 or SEQ ID No: 2. The application of the beta-glucosidase is that the beta-glucosidase is applied to simultaneous saccharification and fermentation production of ethanol from cellulose. The beta-glucosidase (Bgl2350 and Bgl5676) provided by the invention has the characteristics of low temperature and high activity, the optimal reaction temperatures are respectively 40 DEG C and 30 DEG C, and both the beta-glucosidase and the Bgl5676 have excellent glucose and xylose tolerance and can tolerate 600mM glucose and 2M xylose environments. Wherein the enzyme activity of the Bgl2350 is relatively high under a low-temperature condition; the Bgl5676 shows the stability and the salt tolerance which are obviously superior to those of the traditional GH1 family beta-glucosidase. Compared with the commercially available Novozymes Cellic CTec3 commercial enzyme, the Bgl2350 and the Bgl5676 show more prominent application potential in the field of low-temperature cellulose saccharification, and can partially replace the beta-glucosidase component in the commercial enzyme in practical application.

The invention discloses geobacillus thermoglucosidase and application thereof, and belongs to the technical field of functional microorganisms and oil exploitation. The geobacillus thermoglucosidase is preserved in the China General Microbiological Culture Collection Center (CGMCC), and the preservation number of the geobacillus thermoglucosidase is CGMCC No. 32470. The strain can degrade saturated hydrocarbon and asphaltene in the wax-containing thickened oil, and also can generate a biological emulsifier, reduce the viscosity and wax content of wax-containing crude oil and improve the fluidity of the wax-containing thickened oil, so that the deposition of asphaltene and paraffin is delayed and reduced, and the exploitation of the wax-containing thickened oil is facilitated; therefore, the method has good application prospects and values in the technical fields of functional microorganisms and oil exploitation.

The invention provides a method for preparing ginsenoside CK through a multi-enzyme system, and relates to the field of biological enzyme method preparation of active substances. The method comprises the following steps: constructing genetically engineered bacteria capable of heterologously expressing two glucosidase, wherein a synthetic gene sequence of glucosidase A is selected from a gene sequence of glucosidase of Alaska sphingosine box bacteria; the synthetic gene sequence of the glucosidase B is selected from the gene sequence of glucosidase of paenibacillus mucilaginosus; the glucosidase A is used for converting Rb1, Rb2 and Rc in a substrate into Rd and converting Rd into F2, and the glucosidase B is used for converting F2 into CK; after two recombinant bio-enzymes are obtained through fermentation, a raw material containing diol ginsenoside is used as a substrate for fermentation and enzymolysis; and heating, filtering, removing enzyme, cooling and crystallizing. The method is low in raw material dependence, high in utilization rate, short in conversion time, high in conversion efficiency, high in product purity after conversion, free of separation and purification operation and suitable for large-scale industrial production.

A composition for administration to a subject is disclosed. The composition comprises at least one plant active component. The plant active component is present in the composition in an amount effective to inhibit alpha-glucosidase in the subject. The plant active component comprises at least one extract of Lysidice rhodostegia. The composition can be in the form of an oral composition, for example for ingestion. The composition can also be in the form of a topical composition, for example for topical administration. A method of inhibiting alpha-glucosidase in a subject is also provided. The method comprises administering to the subject an effective amount of the composition.

The present application provides for compositions comprising high concentrations of acid α-glucosidase in combination with an active site-specific chaperone for the acid α-glucosidase, and methods for treating Pompe disease in a subject in need thereof, that includes a method of administering to the subject such compositions. The present application also provides methods for increasing the in vitro and in vivo stability of an acid α-glucosidase enzyme formulation.

Recombinant human alpha glucosidase (rhGAA) compositions derived from CHO cells are disclosed, comprising a more optimal glycan composition consisting of rhGAA with higher amounts of N-glycans bearing mannose-6-phosphate (M6P) or bis-M6P, along with low amounts of non-phosphorylated high mannose glycans, than conventional rhGAA, along with low amounts of terminal galactose on complex oligosaccharides. Compositions comprising the rhGAA are described, as are methods of use.

The invention discloses beta-glucosidase GE000505 for saponin conversion and a preparation method and application thereof, and belongs to the technical field of biology, the amino acid sequence of the beta-glucosidase GE000505 is shown as SEQ ID NO.1, and the beta-glucosidase GE000505 is coded by a gene with the nucleotide sequence shown as SEQ ID NO.2. The invention further discloses a preparation method of the beta-glucosidase GE000505 for saponin conversion and application of the beta-glucosidase GE000505. The beta-glucosidase GE000505 disclosed by the invention can be used for converting high-content ginsenoside and notoginsenoside into rare saponins R2, Rg3, Rg2 and Rh1, completely converting notoginsenoside R1 into R2, completely converting ginsenoside Rd into Rg3, completely converting ginsenoside Re into Rg2, completely converting ginsenoside Rg1 into Rh1, completely converting ginsenoside F1 into aPPT, completely converting ginsenoside F2 into aPPD, and completely converting ginsenoside F2 into aPPD. The problem that existing beta-glucosidase is not resistant to alcohol in the process of converting ginsenoside is solved.

The present disclosure provides engineered acidic alpha-glucosidase polypeptides, recombinant polynucleotides encoding the engineered acidic alpha-glucosidase polypeptides, and methods of using the engineered acidic alpha-glucosidase polypeptides and the recombinant polynucleotides for therapeutic purposes.

A method of producing an alcohol or alcohol precursor from a cellulosic material and a genetically modified ethanologenic organism. The genetically modified organism comprises at least one endoglucanase (cen-like), an exoglucanase (cex-like) or beta-glucosidase 1 (bgl1).

The present application provides a Monosporozyma servazzii, which is a Monosporozyma servazzii AMCC 31719 strain, which is preserved in the China Center for Type Culture Collection (CCTCC) and has a preservation number of CCTCC NO: M 20242063. The Monosporozyma servazzii strain of the present application can utilize multiple carbon sources during growth, has the characteristics of high glycerol production, beta-glucosidase production and aroma production, can improve the flavor of baked products and improve the texture, and is mainly applied in the field of food or fermented food.

A transformant obtained by introducing (a) a lactate dehydrogenase gene and / or (b) a malate / lactate dehydrogenase gene into a Hydrogenophilus bacterium efficiently produces lactic acid through use of carbon dioxide as a sole carbon source. Parageobacillus thermoglucosidasius ldh gene, Geobacillus kaustophilus ldh gene and Thermus thermophilus ldh gene of lactate dehydrogenases, and Thermus thermophilus mldh gene and Meiothermus ruber mldh-1 and mldh-2 genes of malate / lactate dehydrogenases are preferable in that they have good lactic acid production efficiency.