30 results about "Hyaluronan synthase" patented technology

Disclosed are DNA segments encoding <DEL-S DATE="20010821" ID="DEL-S-00001">hyaluronic acid synthase which are employed to construct recombinant cells useful in the production of hyaluronate synthase or hyaluronic acid (HA)<DEL-E ID="DEL-S-00001"> <INS-S DATE="20010821" ID="INS-S-00001">the recombinant DNA segment identified in FIG. 5. <INS-E ID="INS-S-00001">In preferred aspects, chromosomal DNA from Streptococcus equisimilis is partially digested with EcoRI and the resultant fragments are ligated to form recombinant vectors. These vectors are useful in the transformation of host cells such as E. coli and or Streptococcal hosts. <DEL-S DATE="20010821" ID="DEL-S-00002">Resultant transformants are screened by the novel screening assays to identify colonies which have incorporated HA synthase DNA in a form that is being actively transcribed into the corresponding HA synthase enzyme. These colonies may be selected and employed in the production of the enzyme itself or its product, HA.<DEL-E ID="DEL-S-00002"> <INS-S DATE="20010821" ID="INS-S-00002">The recombinant DNA segment identified in FIG. 5 is then inserted into a recombinant Streptococcal host for the production of hyaluronic acid (HA).<INS-E ID="INS-S-00002">

The present invention relates to a recombinant Bacillus host cell containing a recombinant vector including a nucleic acid segment having a coding region segment encoding enzymatically active hyaluronan synthase (HAS). The recombinant Bacillus host cell is utilized in a method for producing hyaluronic acid (HA).

The present invention relates to methods for producing a hyaluronic acid with a desired average molecular weight in the range of 20,000-800,000 Dalton, the methods comprising the steps of: (a) cultivating a recombinant Bacillus host cell at a first temperature conducive to its growth, wherein the Bacillus host cell comprises a nucleic acid construct comprising a hyaluronan synthase encoding sequence operably linked to a promoter sequence foreign to the hyaluronan synthase encoding sequence; (b) then cultivating the recombinant Bacillus host cell of step (a) at a second temperature higher than the first temperature of step (a) under conditions suitable for production of the hyaluronic acid, whereby the Bacillus host cell produces hyaluronic acid with a desired average molecular weight in the range of 20,000-800,000 Dalton; and (b) recovering the hyaluronic acid.

The present invention relates to methodology for polymer grafting by a polysaccharide synthase and, more particularly, polymer grafting using the glycosaminoglycan synthases from Pasteurella multocida. The methodology of the present invention includes providing an enzymatically active glycosaminoglycan synthase enzyme from Pasteurella multocida, providing a synthetic, artificial acceptor for the glycosaminoglycan synthase enzyme; combining the synthetic, artificial acceptor with the glycosaminoglycan synthase enzyme within a reaction medium, wherein the reaction medium contains at least one sugar precursor selected from the group consisting of UDP-GlcA, UDP-GlcNAc, UDP-GalNAc, and reacting the glycosaminoglycan synthase enzyme with the synthetic, artificial acceptor to produce an oligosaccharide or polysaccharide polymer. The glycosaminoglycan synthase enzyme may be hyaluronan synthase, chondroitin synthase, or heparosan synthase from P. multocida, and the oligosaccharide or polysaccharide polymer may be hyaluronic acid (hyaluronan), chondroitin, heparosan, or combinations thereof.

The present invention relates to a nucleic acid segment having a coding region segment encoding enzymatically active Pasteurella multocida hyaluronate synthase (PmHAS), and to the use of this nucleic acid segment in the preparation of recombinant cells which produce hyaluronate synthase and its hyaluronic acid product.

The invention discloses a method for producing hyaluronic acid with different molecular weights through reasonable regulation, belonging to the technical field of bioengineering. The method comprises the following steps: integrating hyaluronic synthase hasA from streptococcus zooepidemicus on a bacillus subtilis genome; performing modularized assembly and expression analysis on genes of UDP-GlcA and UDP-GlcNAc synthetic routes in bacillus subtilis, and producing hyaluronic acids with different molecular weights from the bacillus subtilis by controlling different UDP-GlcA and UDO-GlcNAc concentrations. The method disclosed by the invention can be used for laying a certain foundation for efficiently preparing the hyaluronic acids within a specified molecular weight range and is suitable for industrial production and application.

The present invention relates to methodology for polymer grafting by a polysaccharide synthase and, more particularly, polymer grafting using the hyaluronate synthase from Pasteurella multocida. The present invention also relates to coatings for biomaterials wherein the coatings provide protective properties to the biomaterial and / or act as a bioadhesive. Such coatings could be applied to electrical devices, sensors, catheters and any device which may be contemplated for use within a mammal. The present invention further relates to drug delivery matrices which are biocompatible and may comprise combinations of a biomaterial or a bioadhesive and a medicament or a medicament-containing liposome. The biomaterial and / or bioadhesive is a hyaluronic acid polymer produced by a hyaluronate synthase from Pasteurella multocida. The present invention also relates to the creation of chimeric molecules containing hyaluronic acid or hyaluronic acid-like chains attached to various compounds and especially carbohydrates or hydroxyl containing substances. The present invention also relates to a chondroitan synthase from Pasteurella multocida which is capable of producing polysaccharide polymers on an acceptor or primer molecule.

The invention discloses a recombinant Pichia pastoris for producing small-molecular hyaluronic acids and a construction method thereof, belonging to the technical field of bioengineering. Hyaluronic acid synthase hasA derived from Streptococcus zooepidemicus and UDP-glucose dehydrogenase tuaD derived from Bacillus subtilis are expressed in recombinant Pichia pastoris GS115 host to implement production of hyaluronic acids. Meanwhile, hyaluronidase derived from hirudo is integrated to the Pichia pastoris genome and subjected to secretion expression by constitutive promoters with different intensities; the secretion expression level of the hyaluronidase is controlled to prepare small-molecular hyaluronic acid products with different molecular weights; and the prepared products have different ranges of molecular weight, and have instruction and reference meanings for directly producing small-molecular hyaluronic acids within a specified range from microbes. The recombinant Pichia pastoris lays certain foundation for high-efficiency preparation of small-molecular hyaluronic acids, and is suitable for industrial production.

A plasmid, recombinant engineering bacteria and a preparation method of hyaluronic acid with uniform molecular weight belong to the technical field of gene engineering. A first expression plasmid containing a Pasteurella multocida hyaluronan synthase gene (SEQ NO.1 in the sequence table) and a second recombinant expression plasmid containing hyaluronic acid precursor synthesis related genes (SEQ NO.2, SEQ NO.3, SEQ NO.4 and SEQ NO.5 in the sequence table) cloned from Bacillus subtilis are introduced into gram positive safe microorganisms, and an expression regulation and control system consisting of the two recombinant expression plasmids respectively controls hyaluronan synthase and hyaluronic acid precursor synthesis related enzyme, so as to obtain hyaluronic acid with uniform molecular weight by a biosynthesis method. The method provided by the invention not only improves the capability of the host strains to synthesize hyaluronic acid, but also can obtain several varieties of sufficient pure hyaluronic acid with uniform molecular weight.

The present invention relates to a nucleic acid segment having a coding region segment encoding enzymatically active Streptococcus equisimilis hyaluronate synthase (seHAS), and to the use of this nucleic acid segment in the preparation of recombinant cells which produce hyaluronate synthase and its hyaluronic acid product. Hyaluronate is also known as hyaluronic acid or hyaluronan.

The invention provides a moisturizing mask and a preparation method thereof. The moisturizing mask comprises the following components, by weight: 0.01 to 1% of saccharide isomeride, 0.01%-1% of rhizobium gum, 0.01%-1% of tremella extract, 0.01%-0.2% of sodium hyaluronate, 0.01%-1% of codium tomentosum extract, 0.04%-1.5% of a skin conditioner, 0.1%-0.3% of a pH regulator, 0.06%-0.45% of a thickener and the balance being water. According to the moisturizing mask, various moisturizing active matters are mixed based on a scientific proportion; gene expression of filaggrin, loricrin, hyaluronan synthase-3 and acid sphingomyelinase in the skin can be up-regulated and the water locking capacity of the skin can be improved; meanwhile, synthesis of ceramide and cerebroside is promoted, absorptionof the skin to surrounding moisture is improved, so that the purpose of efficient moisturizing is achieved.

The invention discloses a hyaluronan synthase mutant and application thereof and belongs to the technical field of bioengineering. The mutant with positive effect in both yield and molecular weight is acquired by subjecting hyaluronan synthase of Streptococcus zooepidemicus origin to intracellular domain mutation in Bacillus subtilis; meanwhile, genes of UDP-GlcUA and UDP-GlcNAc synthetic routes in Bacillus subtilis are subjected to modular assembly expression analysis, and high yield of hyaluronan with wide molecular weight range from Bacillus subtilis is implemented by controlling different UDP-GlcA and UDP-GlcNAc concentrations and ratios. Certain basis is laid for the efficient preparation of hyaluronan from food-grade microorganisms, and this mutant is suitable for industrial production and application.

A functionally active hyaluronan synthase having at least one modified amino acid residue therein as compared to a corresponding functionally active native hyaluronan synthase such that the functionally active hyaluronan synthase has an altered enzymatic activity as compared to the corresponding functionally active native hyaluronan synthase is disclosed. The altered enzymatic activity may result in increased or decreased activity when compared to the corresponding native hyaluronan synthase, or the altered enzymatic activity may result in production of hyaluronan having an average molecular mass that is greater than or less than an average molecular mass of hyaluronan produced by the corresponding native hyaluronan synthase. Methods of producing hyaluronic acid utilizing a recombinant host cell having an expression construct encoding the functionally active hyaluronan synthase with altered enzymatic activity are also disclosed.

The present invention relates to a recombinant Bacillus host cell containing a recombinant vector including a nucleic acid segment having a coding region segment encoding enzymatically active hyaluronan synthase (HAS). The recombinant Bacillus host cell is utilized in a method for producing hyaluronic acid (HA).

The present invention relates to a nucleic acid segment having a coding region segment encoding enzymatically active Streptococcus equisimilis hyaluronate synthase (seHAS), and to the use of this nucleic acid segment in the preparation of recombinant cells which produce hyaluronate synthase and its hyaluronic acid product. Hyaluronate is also known as hyaluronic acid or hyaluronan.

The invention provides an engineering strain for producing high-molecular-weight hyaluronic acid, a construction method thereof and application of the engineering strain in producing the high-molecular-weight hyaluronic acid. The engineering strain is bacillus subtilis, and the Latin scientific name of the engineering strain is Bacillus subtilis; the engineering strain is named as BH-2, and the preservation number of the strain is CGMCC No.16841; the engineering strain selects bacillus subtilis WB600 as a basic strain, a hyaluronic synthase hasA gene, a UDP-glucose dehydrogenase has B gene anda UDP-glucose pyrophosphorylase hasC gene are introduced into the genome of the engineering strain, the characteristic of the low protease activity of the engineering strain is maintained, the growthand metabolism of the engineering strain are controlled through temperature, the high-molecular-weight hyaluronic acid with the molecular weight of 6.9*106 Dalton is obtained, and the engineering strain has industrial application value.

Disclosed are DNA sequences encoding hyaluronic acid synthase that are employed to construct recombinant cells useful in the production of hyaluronate synthase and hyaluronic acid (HA). In preferred aspects, chromosomal DNA encoding the HA synthase gene, hasA, was cloned from a Streptococcus pyogenes genomic library. These vectors were used to transform host cells such as E. coli and acapsular Streptococci to produce hyaluronic acid. Resultant transformants were screened to identify colonies which have incorporated HA synthase DNA in a form that is being actively transcribed into the corresponding HA synthase enzyme. These colonies were selected and employed in the production of hyaluronic acid.

A functionally active hyaluronan synthase having at least one modified amino acid residue therein as compared to a corresponding functionally active native hyaluronan synthase such that the functionally active hyaluronan synthase has an altered enzymatic activity as compared to the corresponding functionally active native hyaluronan synthase is disclosed. Methods of producing hyaluronic acid utilizing a recombinant host cell having an expression construct encoding the functionally active hyaluronan synthase with altered enzymatic activity are also disclosed.

A functionally active hyaluronan synthase having at least one modified amino acid residue therein as compared to a corresponding functionally active native hyaluronan synthase such that the functionally active hyaluronan synthase has an altered enzymatic activity as compared to the corresponding functionally active native hyaluronan synthase is disclosed. Methods of producing hyaluronic acid utilizing a recombinant host cell having an expression construct encoding the functionally active hyaluronan synthase with altered enzymatic activity are also disclosed.

The invention discloses a Bacillus subtilis self-regulating expression system and its construction method and application, belonging to the field of genetic engineering. The expression system of the present invention comprises a bacillus subtilis that expresses an endonuclease gene induced by xylose and a plasmid expression vector that constitutively expresses an endonuclease gene. In the culture medium containing the inducer xylose, only the Bacillus subtilis with the stable expression vector can grow normally. Fluorescent protein gene (gfp), hyaluronic acid synthase gene (hasA) and keratinase gene (ker) were respectively expressed by the expression system of the present invention, and by fermentation and cultivation, the fluorescence intensity, hyaluronic acid content and keratinase activity were compared with Plasmids maintained by antibiotics increased by 21.8%, 45% and 30.2%.

The invention relates to use of plant fermented extracts, especially relates to the use of imperata cylindrica fermented extract for enhancing the gene expression of keratin, filaggrin and hyaluronansynthase, promoting the proliferation of collagen and elastin, and enhancing antioxidant capacity of skin cells. The invention provides the imperata cylindrica fermented extract by performing two-stage fermentation on an imperata cylindrica water extract with saccharomycetes and lactic acid bacillus. The invention further provides the uses of the imperata cylindrica fermented extract for preparing a composition enhancing the expression quantity of keratin, filaggrin and hyaluronan synthase, promoting the proliferation of collagen and elastin, and enhancing antioxidant capacity of skin cells.The imperata cylindrica fermented extract can effectively improving the moisturizing capacity of skin, maintaining the arrangement of keratinocyte and the completeness of cuticle structure, thus improving the shield function of skin, making skin compact and elastic, and preventing aging of skin.

Provided for are three novel isoenzyme variants of the human Hyaluronan Synthase gene, HAS1Va, HAS1Vb, and HAS1Vc. Also provided is a method for assessing disease, or susceptibility to disease, in a patient; in which a cell or sample of a cell population is mixed with at least one compound which specifically binds to HAS1Va, HAS1Vb, HAS1Vc or HAS2 isoenzyme or isoenzyme variant genomic mRNA transcript or products arising therefrom; and methods of treatment.