39 results about "Phenylmethylsulfonyl Fluoride" patented technology

The invention provides a method for improving the expression quantity of a pyrethroid degeneration enzyme. The method comprises the following steps: preparing and producing strain escherichia coli (E.coli) BLR (DE3) / pET-28a-est825; culturing seeds; carrying out fermentation culture: cooling a mixed induction solution of lactose and isopropylthio-galactoside (IPTG) (at the molar concentration ratio of 7 to 3) to 30 DEG C to 35 DEG C and starting to induce, wherein a mixed material supplementing solution of glycerin and yeast powder is continually added in a flowing way in an induction process, the concentration of the glycerin in a fermentation solution is kept to be 1g / L to 2g / L, and induction time lasts for 6h to 7h; finally, preparing the strain into a fungal suspension solution with the mass concentration of 15 percent to 20 percent by utilizing a phosphate buffering solution containing 0.005mol / L of phenylmethylsulfonyl fluoride and having the pH (Potential of Hydrogen) of 6.5 to 7.0, and carrying out ultrasonic crushing, centrifuging and freeze-drying to obtain high-enzyme-activity degeneration enzyme powder.

The invention relates to a high-temperature Daqu leaching liquor macro-protein extraction and purification method. The method comprises the following steps: crushing high-temperature Daqu, spraying sterile water and evenly mixing, storing in a biological culture tank, performing activated culturing for later use by adopting a stage temperature rise method; adding an acetic acid-sodium acetate buffer solution and a phenylmethylsulfonyl fluoride solution, shaking and blending evenly, and then leaching overnight, filtering and centrifuging to obtain high-temperature Daqu leaching liquor; sequentially adding a TCA acetone solution, an ammonium acetate methanol solution, an acetone solution, a Tris satured phenol / lauryl sodium sulfate buffer solution, an ammonium acetate methanol solution, and a methanol and acetone solution to obtain sediment A, sediment B, phenol layer, sediment C, sediment D and high-temperature Daqu macro-protein samples; adding a sample lysate, performing ice bath ultrasonic for hydrotropy, and centrifuging the sediments to obtain a high-temperature Daqu macro-protein sample solution. The high-temperature Daqu macro-protein sample solution prepared by adopting the method can be applied to two-dimensional electrophoresis of the high-temperature Daqu macro-protein, to obtain high-resolution and high-repeatability two-dimensional electrophoretogram.

The invention discloses a method for keeping phytoferritin stable, and belongs to the technical field of stabilizing functional food. Phenylmethylsulfonyl fluoride is added to a phytoferritin solution, so that the concentration of the solution is between 0.1 and 9 mmol / L to inhibit the degradation of the phytoferritin. The addition of the phenylmethylsulfonyl fluoride can keep good stability of the phytoferritin, so the function of iron supplementation activity is exerted effectively; and the method also increases the yield of protein, prolongs the storage time for the protein, and is favorable for wide application of the phytoferritin.

The invention discloses a liquid chromatography-mass spectrometry measuring method of an acetyl coenzyme A in animal livers. The method specifically comprises the steps that firstly, the fresh animallivers are taken, and blood in the animal livers is washed off; then the liver sample is mixed with pretreatment liquid, homogenizing is carried out, and thus tissue homogenate is prepared, wherein the pretreatment liquid is a perchloric acid aqueous solution containing dithiothreitol and phenylmethylsulfonyl fluoride; an internal standard substance is added into the homogenate and mixed evenly, then centrifugation is carried out, and supernatant is taken as a sample to be detected; and finally, the sample is sent to a liquid chromatography-mass spectrometry detector to be detected, data are collected, and after standard curve regression calculation, the concentration of the acetyl coenzyme A in the animal livers is obtained. Compared with an existing enzyme-linked immunoassay labeling method, the liquid chromatography-mass spectrometry method adopted by the invention is rapid, reliable and simple in operation step and suitable for measuring the content of the acetyl coenzyme A in thelivers in batches.

The invention discloses an extraction method of non-denatured protein and extraction liquid specially used by the method. The extraction method comprises a step a2): extracting the non-denatured protein in a sample by adopting the extraction liquid, wherein the extraction liquid contains sucrose, NaCl, L-sodium ascorbate, digitonin, dithiothreitol, phenylmethylsulfonyl fluoride and a protease inhibitor. The extraction method provided by the invention is easy and convenient to operate, a reagent is mild and economic, high-quality non-denatured protein can be efficiently extracted, the method lays a foundation and provides technical support for subsequently performed adverse plant proteomics research, and the method has a certain guiding significance for extracting recalcitrant tissue protein of plants living in an extreme environment. The method has a significant application value.

The invention discloses an extraction method and application suitable for obtaining a large amount of high-purity Salmonella outer membrane protein. The method comprises the following steps: (1) cloning the gene of Salmonella outer membrane protein into an expression vector, then introducing the obtained recombinant expression vector into host Salmonella to obtain a recombinant strain; (2) expanding and culturing the recombinant strain, and then Collect the cells by centrifugation and cryopreserve to obtain the frozen cells; (3) add protein lysate and phenylmethylsulfonyl fluoride solution to the frozen cells, and ultrasonically break at constant temperature to collect protein precipitates; (4) add protein precipitates Add the outer membrane protein extract to the mixture, mix well and incubate at 4°C, then centrifuge to collect the supernatant; (5) separate and purify the supernatant to obtain the Salmonella outer membrane protein. The method of the invention has simple steps and convenient operation, is suitable for large-scale extraction of high-purity and high-activity outer membrane proteins, and can simultaneously meet the requirements of follow-up experiments such as protein activity, structure, and vaccine research and development.