| Panax notoginseng flower as a dual-use food,not only has a variety of Pharmacological activities and edible value,and has attracted wide attention of researchers.At present,the research on Panax notoginseng flower is mainly focused on the saponins and their biological activities,while the studies on the protein composition and functional properties of the flower of Panax notoginseng are few.Therefore,this paper used Panax notoginseng flower as raw material,used Osborne method to extract different kinds of protein;single factor experiment was used to optimize the extraction conditions of Panax notoginseng flower Albumin and Globulin.Ultrafiltration,ion exchange chromatography and gel filtration chromatography were used to separate and purify Panax notoginseng flower Albumin and Globulin.The structure was identified by SDS-PAGE and LC-MS/MS analysis.The physical and chemical properties and functional properties of Panax notoginseng flower Albumin and Globulin were determined by Fourier transform infrared spectroscopy(FTIR),and the anticoagulant activities of different kinds of Panax notoginseng flower proteins were preliminarily discussed,which providedtheoretical basis for the application of Panax notoginseng flower in food industry.The main contents and conclusions are as follows:1.Analysis of protein content in different parts of Panax notoginsengThe content of crude protein in the taproot,stem leaf and flower of Panax notoginseng respectively were 4.81%,11.18% and 18.47%,and the content of crude protein in the flower of Panax notoginseng was the highest.The contents of albumin,globulin,gluten and gliadin in pantoginseng respectively were 6.47%,3.04%,0.88%and 1.46%,and albumin and globulin were higher.Therefore,the content of serum and globulin of Panax notoginseng Flower was used as the index to study the extraction technology of Panax notoginseng Flower Protein.2.Study on the extraction technology of Panax notoginseng flower Albumin and GlobulinThe optimal extraction conditions were as follows: solid-liquid ratio of 1:40,extraction temperature of 30?,extraction time of 90 min,extraction rate of 39.02%,purity of 87.41%.The optimal extraction conditions of Panax notoginseng globulin were as follows: liquid to solid ratio of 1:30,extraction temperature of 40?,extraction time of 90 min,extraction rate of 4.61%,purity of 68.44%.3.Isolation,purification and identification of serum and globulin from Panax notoginsengThree elution peaks A1 ? A2 and A3 were isolated and purified by DEAE.The molecular weights of the components were measured by SDS-PAGE,which respectively were about 33 k Da?37k Da and 90 k Da.Sephadex G-50 was used to further purify the elution peak A1,and a single protein elution peak PNFA-1 with a molecular weight of 37 k Da was obtained.Two elution peaks B1 and B2 were isolated and purified by DEAE.The molecular weights of B1 and B2 were determined by SDS-PAGE,which respectively were about 65 k Da and 37 k D.Sephadex G-50 was used to further purify the elution peak B2,and a single protein elution peak PNFA-2 with a molecular weight of 65 k Da was obtained.4.Study on physicochemical properties of albumin and globulin of Panax notoginsengThe isoelectric points of Panax notoginseng albumin and globulin were 3.8 and4.7 respectively.The secondary structure of albumin and globulin of Panax notoginseng was mainly ?-fold,the proportion of which was 51.2% and 47.4%,respectively.The total phobicity contents of Panax notoginseng albumin and globulin were 9.83?mo L/g and 9.34?mo L/g,respectively.The free phobicity was 5.91 ?mo L/g and 7.62?mo L/g,respectively.The disulfide bond was 3.92?mo L/g and 1.72?mo L/g,respectively.The surface hydrophobicity of Panax notoginseng albumin and globulin was 699.83±20.6 and 1613.9±19.8,respectively.The total amino acids of Panax notoginseng albumin and globulin were 93.43mg/g and 155.423mg/g,respectively,and the contents of essential amino acids respectively were 20.89mg/g and51.82mg/g.Different drying methods have certain effects on the physicochemical properties of Panax notoginseng albumin and globulin.Freeze-drying and hot-air drying at 80?showed a slightly higher content of free sulfhydryl groups in Panax notoginseng albumin and globulin,indicating that disulfide bond fracture and rearrangement occurred in the treatment process.Under freeze-drying conditions,the surface hydrophobicity of Panax notoginseng albumin and globulin was the highest,while under hot air drying at 80?,the surface hydrophobicity of protein decreased.The secondary structure of Panax notoginseng protein was further analyzed.It was found that the content of Panax notoginseng albumin and globulin ?-folded structure increased significantly with the increase of temperature.These results indicate that the thermal denaturation of Panax notoginseng protein can break the intermolecular hydrogen bonds and cause significant changes in protein structure.5.Study on functional properties of albumin and globulin of Panax notoginsengThe water holding capacity of Panax notoginseng albumin and globulin were4.827g/g and 3.795g/g,respectively.The albumin had stronger water soluble ability and water absorption than the globulin.The foaming ability of Panax notoginseng albumin and globulin was 12.12% and 6.25%,respectively.The foaming ability of albumin was stronger than that of globulin.The emulsifying ability of albumin in Panax notoginseng flower is better than that of globulin,but the emulsifying stability of globulin is better.Different drying methods have certain effects on the functional properties of Panax notoginseng albumin and globulin.With the increase of drying temperature,the solubility,emulsification and foaming ability of Panax notoginseng albumin and globulin decreased,while emulsification stability and turbidity increased.6.Study on anticoagulant biological activity of various proteins of Panax notoginsengCrude protein of different parts of Panax notoginseng root,stem,leaf and flower all had anticoagulant effect,among which the anticoagulant effect of Panax notoginseng flower crude protein was the most significant.At the concentration of10mg/m L,Panax notoginseng albumin,globulin and gliadin could significantly prolong the time of PT?TT and APTT,and globulin had the best anticoagulant effect. |
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