34 results about "Phospholipase C" patented technology

Indole and indole-related compounds, compositions and methods are disclosed. The compounds of the invention are useful as phospholipase inhibitors. The compounds and compositions of the invention are useful for treatment of phospholipase-related conditions, such as insulin-related, weight-related and / or cholesterol-related conditions in an animal subject.

A method for degumming an oil composition comprises the steps of (a) providing an oil composition containing a quantity of phospholipids, (b) contacting said oil composition simultaneously with one or more phospholipase A enzymes and one or more phospholipase C enzymes, under conditions sufficient for the enzymes to react with the phospholipids to create phospholipid reaction products, and (c) separating the phospholipids reaction products from the oil composition, the remaining oil composition after the separation being a degummed oil composition, whereby during step (b) the reaction of said one or more phospholipase A enzymes proceeds at a faster rate than it would in the absence of said one or more phospholipase C enzymes.

The present invention relates to inositol phosphate derivatives, in which the inositol phosphate is substituted with one or two reactive groups G or one or two conjugated substances or molecules M, said reactive group(s) G or said substance(s) or molecule(s) M being linked to IP1 via a linkage group L, M being chosen from the following group: a tracer, an immunogen, a member of a binding partner pair, a solid support.Application: tools allowing the study of the inositol phosphate cycle and therefore, indirectly, the study of seven transmembrane domain receptors coupled to phospholipase C, receptors having a tyrosine kinase activity, and in general enzymes involved in the variations of the intracellular concentration of IP1.

Glycosylphosphatidylinositol-(GPI-) anchored HSPG glypican-1 is strongly expressed in human breast and pancreatic cancer—both by the cancer cells and in the case of pancreatic cancer the adjacent fibroblasts—whereas expression of glypican-1 is low in the normal pancreas and in chronic pancreatitis. Treatment of two pancreatic cancer cell lines, which express glypican-1, with the enzyme phosphoinositide-specific phospholipase-C (PI-PLC) abrogated their mitogenic responses to two heparin-binding growth factors: fibroblast growth factor-2 (FGF2) and heparin-binding EGF-like growth factor (HB-EGF). Treatment of MDA-MB-231 and MDA-MB-468 breast cancer cells with PI-PLC abrogates the mitogenic response to two heparin-binding growth factors, heparin-binding epidermal growth factor-like growth factor (HB-EGF) and fibroblast growth factor-2 (FGF-2). Syndecan-1 is also expressed at high levels in breast cancer tissues as well as breast cancer cells by comparison with breast normal tissues. Temporary or permanent transfection of a glypican-1 antisense construct attenuated glypican-1 protein levels and the mitogenic response to FGF2 and HB-EGF. Glypican can be used to detect the carcinoma in vitro and therapeutics that either bind to (e.g., antibodies or drugs), remove (e.g., enzymes) or prevent the expression (e.g., antisense constructs) of surface of the extracellular domain of glypican-1 are effective in retarding the growth of glypican-responsive carcinomas.

The invention relates to a bacterial strain capable of producing phosphatidase C, and a screening method thereof. The bacterial strain is Pseudomonas fluorescens P.f-9103, and preservation number is CCTCC No.M2013298. The bacterial strain is obtained by screening and separation from soil near oil workshops. Morphological characteristics are that: the shape of a bacterial colony is round, the surface of the colony is salient, the color is white, and the bacterial colony is nontransparent and yellows with the increasing of age; the colony is smooth, is relatively viscous and dense, and is easy to be stirred up, and the edge is smooth; the colony has a flagellum, is capable of moving, and produces no blastema; the bacterial strain is gram negative, the optimum growth temperature is 25 to 30 DEG C, and pH value is 7.0 to 7.6. The screening method comprises following steps: primary screening is performed on LB medium containing yolk; then phosphatide is taken as a substrate, and enzymatic reaction is performed; and reaction products are analyzed by HPLC so as to realize secondary screening. According to the method, natural phosphatide is taken as the substrate, so that problems caused by inactivity of phosphatidase C on phosphatide substrate analogue NPPC are avoided. Sources of primary screening raw material are abundant, operation is convenient, secondary screening is accurate and rapid, and the screening method is suitable for high throughput screening of the bacterial strain capable of producing phosphatidase C.

The invention provides an isolated Cladosporium strain expressing phospholipase C, which is preserved in the General Microorganism Center of China Microbiological Culture Collection Management Committee, and the preservation number is CGMCC No.7508. The phospholipase C prepared by using the bacterial strain and the preparation method provided by the invention can be used for oil refining, has good degumming effect, and can be widely used in the fields of oil refining, additives, medicine and the like.

Provided is a method for simply and accurately measuring sphingomyelin in a sample, and a kit therefore. The method is a method for measuring sphingomyelin in a sample, comprising reacting the sample with a phospholipase D which does not react with sphingomyelin and lysophosphatidylcholine but reacts with phosphatidylcholine, a lysophospholipase or a monoglyceride lipase, and a choline oxidase, eliminating the formed hydrogen peroxide, reacting the resultant with a phospholipase D which does not react with glycerol-3-phosphorylcholine and free fatty acid but reacts with sphingomyelin, and a choline oxidase, and measuring the formed hydrogen peroxide.

The invention relates to a new method for measuring phospholipase A1 or A2 activity in a sample, using a solid phase coated with a fluorochrome-labelled phospholipase A1 or A2 substrate, wherein the molecular coverage is in the range from 8 to 30 fluorochrome-labelled phospholipase A1 or A2 substrate molecules / nm2, and kit for carrying out said method.

The invention provides a preservation method of phospholipase C. By using the preservation method, the phospholipase C has good enzyme activity stability. When the preservation method of the phospholipase C provided by the invention is used in the practical application process of the phospholipase C, the production energy consumption can be reduced; and meanwhile, the activity loss of the phospholipase C is avoided.

The invention discloses a recombinant escherichia coli capable of producing phosphatidase C, a phosphatidase C preparation method, and applications of phosphatidase C. A construction method of the recombinant escherichia coli comprises following steps: A, preparation of phosphatidase C gene, wherein PCR primers are designed, and phosphatidase C gene segments are amplified; B, construction of recombinant plasmids, wherein phosphatidase C gene segments with 1158bp are obtained by digestion using EcoR I and Not I enzymes, and the phosphatidase C gene segments are combined with high-copy plasmid pET28a by clone so as to obtain recombinant plasmid Pet28a-P.f-PLC; and C construction of engineering bacteria, wherein plasmid Pet28a-P.f-PLC is transferred into E.coil BL21(DE3) via transformation. Active phosphatidase C can be produced by liquid fermentation, and intracellular and extracellular enzyme activities can reach 408.5U / ml. Phosphatidase C possesses an application prospect in the fields such as oil and fat refining, phosphatide modification and medical treatment.