9 results about "Mannosidase" patented technology

The invention relates to the technical field of traditional Chinese medicine effective component extraction, and provides an efficient extraction and purification process and application of icariin in plateau herba epimedii, the efficient extraction and purification process of icariin in plateau herba epimedii comprises the following steps: S1, epimedium leaves are crushed and sieved to obtain epimedium powder; s2, adding an ethanol solution into the herba epimedii powder, then adding a compound enzyme, performing ultrasonic treatment, inactivating the enzyme, performing suction filtration, and collecting filtrate to obtain an icariin crude extract; s3, diluting the crude icariin extract, adjusting the pH value to 5-6, carrying out chromatographic purification and elution, and collecting the eluate; s4, performing vacuum concentration on the eluent, and performing spray drying to obtain a pure icariin product; the compound enzyme comprises cellulase, pectinase and alpha-mannosidase in a mass ratio of 3: 1: (0.2-1.0). By means of the technical scheme, the problem that in the prior art, the extraction rate of icariin in plateau herba epimedii is low is solved.

The invention provides I-type alpha-mannosidase, an expression vector, and a transgenic cell line and host bacteria containing the expression vector. According to the invention, the function of the SlMNSI1 gene related to tomato protein N-glycosylation is cloned and verified for the first time, heterologous active expression of the SlMNSI1 gene is realized, and the recombinant protein can utilize a high mannose type N-sugar chain (H8N2F0X0) as a substrate to efficiently catalyze and shear mannose residues connected with alpha-1, 2-at the tail end of the recombinant protein so as to generate a Man5GlcNAc2 (H5N2F0X0) product. The invention also provides the SlMNSI1 gene and an encoding protein thereof, an expression vector containing the SlMNSI1 nucleotide sequence, and application of a transgenic cell line and host bacteria containing the expression vector in N-carbohydrate chain modification and glycosylation engineering. The invention further provides application of the expression vector and a transgenic cell line or host bacteria containing the vector in glycoprotein structure optimization, drug research and development and metabolic engineering.

The invention relates to a group of N-glycosylation mutants of type I alpha-mannosidase and application of the N-glycosylation mutants. According to the invention, N-glycosylation modification of tomato-sourced I-type alpha-mannosidase SlMNSI1 at amino acid residues N288 and N334 is analyzed for the first time, and the modification sites are located in a conserved structural domain of a GH47 glycoside hydrolase family. N288A, N334A and N288A / N334A mutants are further constructed, and the N288A, N334A and N288A / N334A mutants are transiently expressed in mesophyll cells of the nicotiana benthamiana. Positioning analysis shows that the mutant can still be partially positioned in the Golgi apparatus, but protein aggregates appear and are transferred to the periphery of the endoplasmic reticulum. Protein stability detection shows that the N-glycosylation modified mutant is obviously accelerated to degrade after being treated by the cycloheximide, which indicates that the glycosylation modification of the N288 and N334 sites has an important effect on the stability and subcellular localization of the SlMNSI1. The functional characteristics of key glycosylation sites of the type I alpha-mannosidase are disclosed for the first time, a group of N-glycosylation mutants with structure and function differences are provided, and the N-glycosylation mutants can be used as important tools for researching plant glycosylation regulation, protein engineering modification and gene function identification and have wide application prospects.

Methods of determining glycan pairing content of a protein composition are provided. In exemplary embodiments, the method comprises (a) treating a sample of the protein composition with (i) a mannosidase and (ii) a β1-4 endoglycosidase, to produce a mixture of components of the protein, wherein at least some of the components comprises one or more digested glycan structures; and (b) separating the components of the mixture based on molecular weight; and (c) quantifying the abundance of glycan pairs, optionally, quantifying (i) paired afucosylated glycans, (ii) unpaired afucosylated glycans, (iii) paired high mannose glycans, and / or (iv) unpaired high mannose glycans, of the mixture. In exemplary aspects, the method comprises quantifying the relative unpaired afucosylated (AF) glycan content and / or relative unpaired high mannose (HM) glycan content of the mixture.

The invention discloses an application of a mannosidase alpha2B enzyme gene MAN2B1 inhibitor in inhibiting physiological behaviors of neurons. The MAN2B1 inhibitor inhibits substance metabolism, adhesion, spreading and protrusion branch growth of the neurons by inhibiting expression of mannosidase alpha2B enzymes. The invention proposes and verifies that the mannosidase alpha2B enzyme gene MAN2B1 inhibitor can effectively inhibit the physiological behaviors of neurons for the first time, and provides a new direction for research and development of drugs for treating diseases caused by excessive growth of neuronal axons of a peripheral or central nervous system.

The present invention relates to an expression system for the extracellular production of recombinant tri-fucosylated N-glycoproteins in insect cells. The invention also relates to an expression cassette comprising a glyco-module comprising polynucleotide sequences of a nematode galactosyltransferase, fucosyltransferase, and mannosidase; and a polynucleotide sequence encoding one or more tri-fucosylated N-glycoproteins derived from a nematode; a recombinant baculovirus comprising said expression cassette, a method for producing said tri-fucosylated N-glycoproteins in transfected insect cells, and a transfected insect cell. The invention also relates to an immunological composition comprising said tri-fucosylated N-glycoproteins, which can be administered to ruminants to induce protective anti-nematode immunity.

The invention belongs to the technical field of molecular biology, and relates to application of alpha-1, 2-mannose structure change in preparation of a liver cancer diagnosis or prognosis product, research is carried out through the effect of alpha-1, 2-mannosidase MAN1C1 in liver cancer, and the result shows that the expression quantity of the MAN1C1 in liver cancer tissue is obviously lower than that in para-carcinoma tissue; low expression of MAN1C1 is related to poor prognosis of the liver cancer patient; lectin MVN detection proves that alpha-1, 2-mannose structures of N-sugar chains in liver cancer tissues are significantly more than paracancerous structures, and high abundance of alpha-1, 2-mannose is related to poor prognosis of liver cancer patients. Further, in-vitro cell experiments show that the survival ability of liver cancer cells is improved under the condition of nutrition deficiency after the MAN1C1 is knocked down, and in-vivo mouse tumorigenesis experiments show that the low expression of the MAN1C1 can promote the growth of tumors. Therefore, the alpha-1, 2-mannose structure can be used as a diagnostic marker for diagnosis and prognosis of liver cancer, and is suitable for liver cancer diagnosis methods such as lectin and the like and liver cancer diagnosis kits.

The invention relates to a herba epimedii extract for activating cell autophagy and a preparation method and application thereof.The herba epimedii extract is prepared from specific zymophyte and compound enzyme, the content of icariside I in the herba epimedii extract is increased, and the herba epimedii extract can activate cell autophagy by up-regulating expression of lysosome mannosidase (LManV). The composition has the advantages that the autophagy flow of cells is obviously enhanced, the fusion of autophagosome and lysosome is promoted, the p62 / Ref( 2) P accumulation caused by the decline of the autophagy function is efficiently reversed, the steady state of the environment in the cells is maintained, the aging speed of the cells is delayed, the support is provided for the aging resistance of the organism from the cellular level, the improvement of the aging-related physical state and the maintenance of the vitality of the organism are helped, and the composition is safe and non-toxic to the human body.

This invention discloses an enzymatic hydrolysate of taro flour with anticoagulant properties. The invention uses taro flour as raw material and prepares the taro flour enzymatic hydrolysate through water extraction, alcohol precipitation, resolution, starch removal with amylase, further alcohol precipitation, further resolution, and glycosidic hydrolysis. The enzymes used are galactosidase, fucosidase, and mannosidase. The taro flour enzymatic hydrolysate prepared by this invention can delay blood clotting time and has anticoagulant properties, and can be used as a functional ingredient to make beverages and foods with anticoagulant effects.