CN105852135A - Preparation method of edible and medicinal fungus protein peptide-ferrous chelate - Google Patents
Preparation method of edible and medicinal fungus protein peptide-ferrous chelate Download PDFInfo
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- CN105852135A CN105852135A CN201610189412.3A CN201610189412A CN105852135A CN 105852135 A CN105852135 A CN 105852135A CN 201610189412 A CN201610189412 A CN 201610189412A CN 105852135 A CN105852135 A CN 105852135A
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- medical fungi
- ferrous
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- 241000233866 Fungi Species 0.000 title claims abstract description 82
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 59
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 59
- 239000013522 chelant Substances 0.000 title claims abstract description 44
- 238000002360 preparation method Methods 0.000 title claims abstract description 26
- 150000001875 compounds Chemical class 0.000 claims abstract description 35
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 24
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 18
- 239000003513 alkali Substances 0.000 claims abstract description 15
- 238000000605 extraction Methods 0.000 claims abstract description 13
- 229960002089 ferrous chloride Drugs 0.000 claims abstract description 13
- NMCUIPGRVMDVDB-UHFFFAOYSA-L iron dichloride Chemical compound Cl[Fe]Cl NMCUIPGRVMDVDB-UHFFFAOYSA-L 0.000 claims abstract description 13
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims abstract description 11
- 108091005804 Peptidases Proteins 0.000 claims abstract description 9
- 239000004365 Protease Substances 0.000 claims abstract description 8
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims abstract description 8
- 238000000034 method Methods 0.000 claims abstract description 8
- 238000003916 acid precipitation Methods 0.000 claims abstract description 7
- 239000011790 ferrous sulphate Substances 0.000 claims abstract description 7
- 235000003891 ferrous sulphate Nutrition 0.000 claims abstract description 7
- 102000004190 Enzymes Human genes 0.000 claims abstract description 6
- 108090000790 Enzymes Proteins 0.000 claims abstract description 6
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims abstract description 6
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims abstract description 6
- 235000011130 ammonium sulphate Nutrition 0.000 claims abstract description 6
- 238000004090 dissolution Methods 0.000 claims abstract description 6
- 235000013305 food Nutrition 0.000 claims abstract description 6
- 108091005658 Basic proteases Proteins 0.000 claims abstract description 5
- 108090000145 Bacillolysin Proteins 0.000 claims abstract description 3
- 102000035092 Neutral proteases Human genes 0.000 claims abstract description 3
- 108091005507 Neutral proteases Proteins 0.000 claims abstract description 3
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 claims description 37
- 239000000243 solution Substances 0.000 claims description 34
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 18
- 244000309464 bull Species 0.000 claims description 17
- 229920001184 polypeptide Polymers 0.000 claims description 17
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 15
- 239000007788 liquid Substances 0.000 claims description 15
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 14
- 240000006794 Volvariella volvacea Species 0.000 claims description 14
- 235000004501 Volvariella volvacea Nutrition 0.000 claims description 14
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 8
- 239000012153 distilled water Substances 0.000 claims description 8
- 238000001556 precipitation Methods 0.000 claims description 8
- 239000012460 protein solution Substances 0.000 claims description 8
- 235000001674 Agaricus brunnescens Nutrition 0.000 claims description 7
- 230000033228 biological regulation Effects 0.000 claims description 7
- 238000006243 chemical reaction Methods 0.000 claims description 7
- 238000002156 mixing Methods 0.000 claims description 6
- 239000007787 solid Substances 0.000 claims description 6
- 238000003453 ammonium sulfate precipitation method Methods 0.000 claims description 5
- 239000000706 filtrate Substances 0.000 claims description 5
- 238000004108 freeze drying Methods 0.000 claims description 5
- SURQXAFEQWPFPV-UHFFFAOYSA-L iron(2+) sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Fe+2].[O-]S([O-])(=O)=O SURQXAFEQWPFPV-UHFFFAOYSA-L 0.000 claims description 5
- 235000001715 Lentinula edodes Nutrition 0.000 claims description 4
- 240000000599 Lentinula edodes Species 0.000 claims description 4
- 235000007685 Pleurotus columbinus Nutrition 0.000 claims description 4
- 240000001462 Pleurotus ostreatus Species 0.000 claims description 4
- 235000001603 Pleurotus ostreatus Nutrition 0.000 claims description 4
- 241001506047 Tremella Species 0.000 claims description 4
- 238000005119 centrifugation Methods 0.000 claims description 4
- 239000012535 impurity Substances 0.000 claims description 4
- 239000008176 lyophilized powder Substances 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 4
- 230000007935 neutral effect Effects 0.000 claims description 4
- 230000006920 protein precipitation Effects 0.000 claims description 4
- 238000010298 pulverizing process Methods 0.000 claims description 4
- 238000003756 stirring Methods 0.000 claims description 4
- 239000006228 supernatant Substances 0.000 claims description 4
- 241000221377 Auricularia Species 0.000 claims description 3
- 241001537207 Flammulina Species 0.000 claims description 3
- 241000222336 Ganoderma Species 0.000 claims description 3
- 240000001080 Grifola frondosa Species 0.000 claims description 3
- 235000007710 Grifola frondosa Nutrition 0.000 claims description 3
- 240000000588 Hericium erinaceus Species 0.000 claims description 3
- 235000007328 Hericium erinaceus Nutrition 0.000 claims description 3
- 241000123113 Phellinus igniarius Species 0.000 claims description 3
- 241001619461 Poria <basidiomycete fungus> Species 0.000 claims description 3
- 238000001035 drying Methods 0.000 claims description 3
- 239000000796 flavoring agent Substances 0.000 claims description 3
- 235000019634 flavors Nutrition 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- 238000005204 segregation Methods 0.000 claims description 3
- 238000010612 desalination reaction Methods 0.000 claims description 2
- 238000000502 dialysis Methods 0.000 claims description 2
- 238000010438 heat treatment Methods 0.000 claims description 2
- 229920000642 polymer Polymers 0.000 claims description 2
- 239000002244 precipitate Substances 0.000 claims description 2
- 238000000751 protein extraction Methods 0.000 claims description 2
- 241000894006 Bacteria Species 0.000 claims 1
- 239000003814 drug Substances 0.000 claims 1
- 238000007710 freezing Methods 0.000 claims 1
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- CWYNVVGOOAEACU-UHFFFAOYSA-N Fe2+ Chemical compound [Fe+2] CWYNVVGOOAEACU-UHFFFAOYSA-N 0.000 abstract description 11
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 abstract description 6
- 150000001413 amino acids Chemical class 0.000 abstract description 5
- 239000013589 supplement Substances 0.000 abstract description 5
- 231100000252 nontoxic Toxicity 0.000 abstract description 3
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- 230000015556 catabolic process Effects 0.000 abstract 1
- 238000006731 degradation reaction Methods 0.000 abstract 1
- 230000002255 enzymatic effect Effects 0.000 abstract 1
- 239000000413 hydrolysate Substances 0.000 abstract 1
- 230000007723 transport mechanism Effects 0.000 abstract 1
- 235000018102 proteins Nutrition 0.000 description 30
- 244000252132 Pleurotus eryngii Species 0.000 description 13
- 235000001681 Pleurotus eryngii Nutrition 0.000 description 13
- 229910052742 iron Inorganic materials 0.000 description 10
- 238000002835 absorbance Methods 0.000 description 5
- 208000015710 Iron-Deficiency Anemia Diseases 0.000 description 4
- 235000001014 amino acid Nutrition 0.000 description 4
- 230000009920 chelation Effects 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 239000012086 standard solution Substances 0.000 description 4
- 239000012085 test solution Substances 0.000 description 4
- DGEZNRSVGBDHLK-UHFFFAOYSA-N [1,10]phenanthroline Chemical compound C1=CN=C2C3=NC=CC=C3C=CC2=C1 DGEZNRSVGBDHLK-UHFFFAOYSA-N 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 3
- -1 hydrolyzes 30-90 min Proteins 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- 230000002459 sustained effect Effects 0.000 description 3
- 241000040710 Chela Species 0.000 description 2
- WTDHULULXKLSOZ-UHFFFAOYSA-N Hydroxylamine hydrochloride Chemical compound Cl.ON WTDHULULXKLSOZ-UHFFFAOYSA-N 0.000 description 2
- YNPNZTXNASCQKK-UHFFFAOYSA-N Phenanthrene Natural products C1=CC=C2C3=CC=CC=C3C=CC2=C1 YNPNZTXNASCQKK-UHFFFAOYSA-N 0.000 description 2
- 206010037660 Pyrexia Diseases 0.000 description 2
- SMWDFEZZVXVKRB-UHFFFAOYSA-N Quinoline Chemical compound N1=CC=CC2=CC=CC=C21 SMWDFEZZVXVKRB-UHFFFAOYSA-N 0.000 description 2
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000004737 colorimetric analysis Methods 0.000 description 2
- 229910001448 ferrous ion Inorganic materials 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 229910052603 melanterite Inorganic materials 0.000 description 2
- 238000011017 operating method Methods 0.000 description 2
- 239000008188 pellet Substances 0.000 description 2
- 239000001632 sodium acetate Substances 0.000 description 2
- 235000017281 sodium acetate Nutrition 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- TUSDEZXZIZRFGC-UHFFFAOYSA-N 1-O-galloyl-3,6-(R)-HHDP-beta-D-glucose Natural products OC1C(O2)COC(=O)C3=CC(O)=C(O)C(O)=C3C3=C(O)C(O)=C(O)C=C3C(=O)OC1C(O)C2OC(=O)C1=CC(O)=C(O)C(O)=C1 TUSDEZXZIZRFGC-UHFFFAOYSA-N 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 1
- 108010082495 Dietary Plant Proteins Proteins 0.000 description 1
- 239000001263 FEMA 3042 Substances 0.000 description 1
- DKKCQDROTDCQOR-UHFFFAOYSA-L Ferrous lactate Chemical compound [Fe+2].CC(O)C([O-])=O.CC(O)C([O-])=O DKKCQDROTDCQOR-UHFFFAOYSA-L 0.000 description 1
- 206010022971 Iron Deficiencies Diseases 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- LRBQNJMCXXYXIU-PPKXGCFTSA-N Penta-digallate-beta-D-glucose Natural products OC1=C(O)C(O)=CC(C(=O)OC=2C(=C(O)C=C(C=2)C(=O)OC[C@@H]2[C@H]([C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)O2)OC(=O)C=2C=C(OC(=O)C=3C=C(O)C(O)=C(O)C=3)C(O)=C(O)C=2)O)=C1 LRBQNJMCXXYXIU-PPKXGCFTSA-N 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000001467 acupuncture Methods 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 150000004697 chelate complex Chemical class 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 208000036654 deficiency anemia Diseases 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000037149 energy metabolism Effects 0.000 description 1
- 239000004222 ferrous gluconate Substances 0.000 description 1
- 235000013924 ferrous gluconate Nutrition 0.000 description 1
- 229960001645 ferrous gluconate Drugs 0.000 description 1
- 235000013925 ferrous lactate Nutrition 0.000 description 1
- 239000004225 ferrous lactate Substances 0.000 description 1
- 229940037907 ferrous lactate Drugs 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 235000012041 food component Nutrition 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000002519 immonomodulatory effect Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 210000004347 intestinal mucosa Anatomy 0.000 description 1
- JEIPFZHSYJVQDO-UHFFFAOYSA-N iron(III) oxide Inorganic materials O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 description 1
- VRIVJOXICYMTAG-IYEMJOQQSA-L iron(ii) gluconate Chemical compound [Fe+2].OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C([O-])=O.OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C([O-])=O VRIVJOXICYMTAG-IYEMJOQQSA-L 0.000 description 1
- 230000007794 irritation Effects 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 231100000957 no side effect Toxicity 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 235000013613 poultry product Nutrition 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000017854 proteolysis Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 239000001117 sulphuric acid Substances 0.000 description 1
- 235000011149 sulphuric acid Nutrition 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 229920002258 tannic acid Polymers 0.000 description 1
- LRBQNJMCXXYXIU-NRMVVENXSA-N tannic acid Chemical compound OC1=C(O)C(O)=CC(C(=O)OC=2C(=C(O)C=C(C=2)C(=O)OC[C@@H]2[C@H]([C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)O2)OC(=O)C=2C=C(OC(=O)C=3C=C(O)C(O)=C(O)C=3)C(O)=C(O)C=2)O)=C1 LRBQNJMCXXYXIU-NRMVVENXSA-N 0.000 description 1
- 229940033123 tannic acid Drugs 0.000 description 1
- 235000015523 tannic acid Nutrition 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J3/00—Working-up of proteins for foodstuffs
- A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
- A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
- A23J3/34—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The present invention belongs to the field of food biotechnology and particularly relates to a preparation method of edible and medicinal fungus protein peptide-ferrous chelate. The preparation method comprises the following steps: an alkali dissolution and acid precipitation method or an ammonium sulfate sedimentation method is used to extract proteins in the edible and medicinal fungi; alkaline protease, neutral protease or compound protease are used to conduct limited enzymatic hydrolysis on the edible and medicinal fungus proteins, the enzymes are deactivated, and edible and medicinal fungus protein enzymatic hydrolysate is prepared; ferrous iron in inorganic ferrous chloride or ferrous sulfate is used to be chelated with the edible and medicinal fungus protein peptides to obtain the edible and medicinal fungus protein peptide-ferrous chelate. The alkali dissolution and acid precipitation method or the ammonium sulfate sedimentation method can significantly improve the extraction rates of the edible and medicinal fungus proteins. By controlling the enzymatic hydrolysis time, the degradation degrees of the proteins can be effectively controlled. The whole preparation technology of the peptide-ferrous chelate is simple. The prepared ferrous-peptide chelate has unique chelating system and transport mechanism, is liable to be absorbed, safe, non-toxic, and low in price, can at the same time supplement amino acids and ferrous iron, and becomes a first choice of a ferrous supplement. The preparation method provides a new idea for the application of the edible and medicinal fungi.
Description
Technical field
The invention belongs to technical field of food biotechnology, be specifically related to the system of a kind of edible and medical fungi protein peptide-ferrous chelate compound
Preparation Method.
Background technology
Ferrous iron is one of trace element of needed by human, and it is to constitute the requisite composition of haemachrome, rises in human body
The effect of conveying oxygen, also participate in propagation and the differentiation of cell, many important physiological process such as energy metabolism, removing toxic substances.According to
World Health Organization, the whole world with the presence of 30% population iron deficiency, iron deficiency anemia is the most universal nutrition in the whole world
Property disease.China's Nutritional Anemia has more than 95% for iron deficiency anemia；Less than 7 years old iron Deficiency Anemia prevalences
About 20%-70%.At present, the preventing and treating of iron deficiency anemia is mainly realized by absorption iron supplementary.Common iron supplementary has sulphuric acid sub-
Ferrum, ferrous chloride, Ferrous gluconate, ferrous lactate etc., although these iron supplementary iron contents are high, iron supplement effect is preferable, but
Owing to they utilization rates in vivo are relatively low, toxic and side effects is big, and has special metal rust taste.Polypeptide chelate ferrum is albumen
The biology iron that the polypeptide that matter hydrolysis generates chelates with ferrous ion and generates, the most not carbonate suspension, tannic acid and fibre
The impact of the interfering materials such as dimension element, is in absorbable divalent state the most all the time, can directly be absorbed by intestinal mucosa cells,
Having no side effect, do not produce any digestive tract irritation, bioavailability is high, is a kind of preferably iron supplementary.
Edible and medical fungi is the higher fungus that a big class has large-scale sporophore, is commonly called as mushroom or gill fungus, wants containing needed by human body
Multiple nutritional components, protein content is the highest, and as the protein content of Flammulina velutiper (Fr.) Sing reaches 20.9%, in Volvariella volvacea (Bull.Ex Franch.) Singer．, protein content reaches
25.9%, substantially exceed vegetable and grain and food that we often eat, also above the protein content of poultry product.Edible and medical fungi is the richest
Containing each amino acid, such as total amino acid content average out to 15.76%(dry weights such as Lentinus Edodes, Pleurotus ostreatus, Tremella), it is necessary to total amino acid content is put down
It is 6.43% (dry weight).Therefore, the polypeptide that the Proteolytic enzyme in edible and medical fungi produces has compared with common animal and vegetable protein
More rich biological activity and higher nutritive value.
Utilize biotechnology to edible and medical fungi (Lentinus Edodes, Auricularia, Pleurotus ostreatus, crab flavour mushroom, agrocyb eaegerita, Poria, Phellinus igniarius (L. ex Fr.) Quel., Volvariella volvacea (Bull.Ex Franch.) Singer．
, Flammulina velutiper (Fr.) Sing, Pleurotus eryngii, Tremella, Caulis Bambusae In Taeniam, Ganoderma, Grifola frondosa and Hericium erinaceus (Bull. Ex Fr.) Pers. etc.) carry out the deep processing of protein resource, give birth to
Screening and the extraction of thing bioactive peptide separate, and then utilize ferrous chloride, ferrous sulfate etc. to chelate, prepare polypeptide-ferrous iron chela
Compound.This edible and medical fungi biologically active polypeptide-ferrous chelate compound can significantly improve absorbance and the utilization rate of ferrum element, and
Also there is antioxidation, antibacterial, immunomodulating, blood fat reducing and blood sugar lowering isoreactivity, have the highest Development volue and application prospect, with
Time can improve the added value of edible and medical fungi, the utilization for edible and medical fungi resource opens new approach, before having wide market
Scape.
Summary of the invention
Present invention aims to the defect that prior art exists, it is provided that a kind of edible and medical fungi protein peptide-ferrous iron chela
The preparation method of compound, has the advantage that technological operation is easy, safety is high.Prepared ferrous iron-peptide chelate complex has uniqueness
Chelating system and transporting mechanism, easily absorbed, safety non-toxic, price are low, can supplement aminoacid and ferrous iron simultaneously.
For achieving the above object, the present invention adopts the following technical scheme that
The preparation method of the edible and medical fungi protein peptide-ferrous chelate compound of the present invention, comprises the steps:
(1) protein extraction: utilize alkali extraction-acid precipitation or ammonium sulfate precipitation method to extract edible and medical fungi albumen；
(2) enzymolysis: use protease that the edible and medical fungi albumen extracted is carried out limited enzymolysis, prepares edible and medical fungi protein peptide and is combined
Thing solution；
(3) prepared by polypeptide-ferrous chelate compound: utilize solid high-temperature mixing method or chelate liquid legal that edible and medical fungi protein peptide is multiple
Polymer solution makes edible and medical fungi protein peptide-ferrous chelate compound；
(4) edible and medical fungi protein peptide-ferrous chelate compound is carried out lyophilization.
Described edible and medical fungi includes: Lentinus Edodes, Auricularia, Pleurotus ostreatus, crab flavour mushroom, agrocyb eaegerita, Poria, Phellinus igniarius (L. ex Fr.) Quel., Volvariella volvacea (Bull.Ex Franch.) Singer．, acupuncture needle
One in mushroom, Pleurotus eryngii, Tremella, Caulis Bambusae In Taeniam, Ganoderma, Grifola frondosa and Hericium erinaceus (Bull. Ex Fr.) Pers. etc..
In step (1), specifically comprising the following steps that first by edible and medical fungi micronizing of described alkali extraction-acid precipitation, will pulverize
After edible and medical fungi mix with the ratio of 1 g:20-60 mL with distilled water, utilize the NaOH of 0.5wt%-2wt% or KOH solution to adjust
Joint pH to 9.0-11.0, first supersound extraction 10-50 min, then extract 60-120 min 50-100 DEG C of water-bath, filter after extraction,
Taking filtrate, filtrate is the edible and medical fungi protein solution that alkali carries, the edible and medical fungi protein solution pH to 2.5-carried with hydrochloric acid regulation alkali
4.0 carry out albumen precipitation, and 3000-5000 r/min is centrifuged 5-10 min and obtains protein precipitation, are washed to neutral recentrifuge, will be heavy
Shallow lake lyophilization i.e. obtains edible and medical fungi albumen.
In step (1), described ammonium sulfate precipitation method specifically comprise the following steps that first by edible and medical fungi micronizing, by powder
Edible and medical fungi after broken mixes with the ratio of 1 g:20-60 mL with distilled water, is gradually added solid ammonium sulfate and makes ammonium sulfate
Saturation reaches 40-60wt%, 3000-5000 r/min and is centrifuged 5-10 min and obtains edible and medical fungi albumen precipitation, utilizes dialysis desalination
Postlyophilization obtains edible and medical fungi albumen.
In step (2), edible and medical fungi protein dissolution is made protein in water by specifically comprising the following steps that of described limited enzymolysis
Amount concentration reaches 1.0-5.0%, regulates pH to 7.0-11.0, and temperature is maintained at 40-60 DEG C, adds the egg of mass concentration 1.0-5.0%
White enzyme, hydrolyzes 30-90 min, enzyme denaturing, obtains edible and medical fungi protein polypeptide complex solution.
In step (3), specifically comprising the following steps that molten for edible and medical fungi protein polypeptide complex of described solid high-temperature mixing method
Liquid is dried prepared lyophilized powder；Ferrous chloride or ferrous sulfate powder and lyophilized powder are mixed according to mass ratio 1:6-5:6,300-
500 DEG C of heating 1-3h prepare edible and medical fungi protein peptide-ferrous chelate compound.
In step (3), what described chelate liquid was legal specifically comprise the following steps that by edible and medical fungi protein polypeptide complex solution with
0.5-2 mol/L ferrous chloride or copperas solution mixing, stir, and regulation pH is 3.0-9.0,30-80 DEG C of condition
Lower sustained response 1.0-2.0 h, cools down after reaction, and the removal of impurity is gone in centrifugation, adds the 75-95% of centrifugal liquid 3-5 times volume
(v/v) ethanol, precipitation stands 12-24h, centrifugal segregation supernatant, it is thus achieved that edible and medical fungi protein peptide-ferrous chelate compound；Wherein,
The volume ratio of ferrous chloride or copperas solution and edible and medical fungi protein polypeptide complex solution is 1:6-5:6.
Protease of the present invention is alkaline protease, neutral protease or compound protease.
Edible and medical fungi protein peptide-ferrous chelate compound that a kind of preparation method as above prepares.
The present invention compared with prior art, has the advantage that
1) present invention utilizes alkali extraction-acid precipitation or ammonium sulfate precipitation method to extract the albumen in edible and medical fungi, and extraction ratio is high；
2) present invention is by controlling edible and medical fungi proteolysis time, obtains the biologically active peptide with high ferrous sequestering activity；
3) by chelating with the ferrous iron in inorganic matter ferrous chloride or ferrous sulfate, organic biologically active peptide-Asia is prepared
Iron chelate has chelating system and the transporting mechanism of uniqueness, is easily absorbed, safety non-toxic, price are low, can supplement amino simultaneously
Acid and ferrous ion；Application for edible and medical fungi provides new approaches, for the organic ferrous supplementary of exploitation and function
Property food theoretical foundation and technical support are provided.
Detailed description of the invention
For the preparation method of the openest present invention, it is illustrated below in conjunction with embodiment.But the invention is not restricted to
Lower embodiment.
Embodiment 1
The preparation method of a kind of Pleurotus eryngii protein peptide-ferrous chelate compound, concretely comprises the following steps:
The first step: by Pleurotus eryngii micronizing, the Pleurotus eryngii after pulverizing mixes with the ratio of 1 g:20mL with distilled water, utilizes
2wt%NaOH regulates pH to 9.0, supersound extraction 10 min, and 60 min, temperature 60 DEG C are extracted in water-bath, use filtered through gauze after extraction,
Filtrate is the Pleurotus eryngii protein solution that alkali carries, and the Pleurotus eryngii protein solution pH to 3.0 carried with hydrochloric acid regulation alkali carries out albumen precipitation,
4000 r/min are centrifuged 5 min and obtain protein precipitation, be washed to neutral recentrifuge, pellet frozen is drying to obtain Pleurotus eryngii egg
In vain, its extraction rate reached 74.16%；
Second step: make albumen quality concentration reach 3.0% in water Pleurotus eryngii protein dissolution, by NaOH solution by Pleurotus eryngii albumen
PH value of solution is adjusted to 10.0, and temperature is maintained at 50 DEG C, adds 3.0% alkaline protease, hydrolyzes 90 min, enzyme denaturing, obtains Fructus Pruni Bao
Mushroom protein polypeptide complex solution；
3rd step: by the volume ratio of solution of ferrous chloride with polypeptide complex solution be by Pleurotus eryngii protein polypeptide complex solution
1:6 ratio mixes with 0.5mol/L solution of ferrous chloride, stirs, and regulates pH6.0, sustained response 1.0 under the conditions of 60 DEG C
H, cools down after reaction, and the removal of impurity is gone in centrifugation, adds the 75%(v/v of 4 times of volumes of centrifugal liquid) ethanol, precipitate standing 12
H, centrifugal segregation supernatant, it is thus achieved that Pleurotus eryngii protein peptide-ferrous chelate compound；
4th step: lyophilization.
The mensuration of ferrum chelation percent: Phen colorimetry
(1) preparation (10 μ g/mL) of ferrum standard solution: accurately weigh 0.0498 g ferrous sulfate (FeSO4·7H2O) it is dissolved in
In 100 mL water, add the 5 dense H of mL2SO4, slight fever drips the KMnO of 2.0wt% after dissolving4Solution, does not takes off to last redness
Till, it is diluted with water to 1000mL, shakes up；
(2) drafting of standard curve: draw standard solution 0,2.0,4.0,6.0,8.0,10.0 mL of 10 μ g/mL ferrum, respectively
It is placed in 50 mL volumetric flasks, adds the oxammonium hydrochloride. l mL of HCl solution 1 mL, 10 wt% of l mol/L, 0.12 wt% adjacent
Phenanthroline 1 mL, is subsequently adding 10 wt% sodium acetate 5 mL, is diluted with water to scale, shakes up；Molten to be not added with the reagent blank of ferrum
Liquid makees reference liquid, measures absorbance, draw standard curve at 510 nm wavelength；
(3) sample determination: the sample accurately weighing 0.05 g is placed in l00 mL beaker, adds 2 mL concentrated hydrochloric acid, treats that sample is complete
After CL, it is settled in l00 mL volumetric flask with distilled water.Accurately absorption 5 mL sample liquid are in 50 mL volumetric flasks, according to
The operating procedure of standard curve measures absorbance.
C in formula: check in the corresponding iron content of sample test solution (μ g) from standard curve；
The quality (g) of M: sample；
V1: the volume (mL) of taken sample test solution during mensuration；
Vo: the constant volume (mL) after sample treatment.
The mensuration of the chelating rate of ferrum:
M in formula1: the amount (mg) of ferrum in chelate；
M0: add the total amount (mg) of ferrum in reaction system.
The chelation percent of the final Pleurotus eryngii protein peptide-ferrous chelate compound measuring preparation is 83.17%.
Embodiment 2
The preparation method of a kind of Volvariella volvacea (Bull.Ex Franch.) Singer． protein peptide-ferrous chelate compound, concretely comprises the following steps:
The first step: by Volvariella volvacea (Bull.Ex Franch.) Singer． micronizing, the Volvariella volvacea (Bull.Ex Franch.) Singer． after pulverizing mixes with the ratio of 1 g:60 mL with distilled water, utilizes
1wt% NaOH regulates pH to 11.0, supersound extraction 50 min, and water-bath is extracted 120 min, temperature 100 DEG C, used gauze after extraction
Filtering, filtrate is the Volvariella volvacea (Bull.Ex Franch.) Singer． protein solution that alkali carries, and the Volvariella volvacea (Bull.Ex Franch.) Singer． protein solution pH to 3.0 carried with hydrochloric acid regulation alkali carries out albumen and sinks
Form sediment, be centrifuged to obtain protein precipitation, be washed to neutral recentrifuge, pellet frozen be drying to obtain edible and medical fungi albumen, its extraction ratio
Reach 83.72%；
Second step: Volvariella volvacea (Bull.Ex Franch.) Singer． protein dissolution making in water albumen quality concentration reach 3.0%, regulates pH to 11.0, temperature is maintained at 40
DEG C, add 3.0 % alkaline proteases, hydrolyze 60 min, enzyme denaturing, obtain Volvariella volvacea (Bull.Ex Franch.) Singer． protein polypeptide complex solution；
3rd step: be 1 by the volume ratio of solution of ferrous chloride Yu polypeptide complex solution by Volvariella volvacea (Bull.Ex Franch.) Singer． protein polypeptide complex solution:
2 ratios mix with 0.5mol/L solution of ferrous chloride, stir, and regulate pH6.0, sustained response 1.0 h under the conditions of 60 DEG C,
Cooling down after reaction, the removal of impurity is gone in centrifugation, adds the 95%(v/v of 5 times of volumes of centrifugal liquid) ethanol, precipitation stands 12 h, centrifugal
Remove supernatant, it is thus achieved that Volvariella volvacea (Bull.Ex Franch.) Singer． protein peptide-ferrous chelate compound；
4th step: lyophilization.
The mensuration of ferrum chelation percent: Phen colorimetry
(1) preparation (10 μ g/mL) of ferrum standard solution: accurately weigh 0.0498 g ferrous sulfate (FeSO4·7H2O) it is dissolved in
In 100 mL water, add the 5 dense H of mL2SO4, slight fever drips the KMnO of 2.0wt% after dissolving4Solution, does not takes off to last redness
Till, it is diluted with water to 1000 mL, shakes up；
(2) drafting of standard curve: draw standard solution 0,2.0,4.0,6.0,8.0,10.0 mL of 10 μ g/mL ferrum, respectively
Being placed in 50 mL volumetric flasks, add the oxammonium hydrochloride. l mL of HCl solution 1 mL, 10wt% of l mol/L, 0.12wt% neighbour is luxuriant and rich with fragrance
Sieve quinoline 1 mL, is subsequently adding 10wt% sodium acetate 5 mL, is diluted with water to scale, shakes up；To be not added with the blank reagent solution of ferrum
Make reference liquid, at 510 nm wavelength, measure absorbance, draw standard curve；
(3) sample determination: the sample accurately weighing 0.05 g is placed in l00 mL beaker, adds 2 mL concentrated hydrochloric acid, treats that sample is complete
After CL, it is settled in l00 mL volumetric flask with distilled water；Accurately absorption 5 mL sample liquid are in 50 mL volumetric flasks, according to
The operating procedure of standard curve measures absorbance；
C in formula: check in the corresponding iron content of sample test solution (μ g) from standard curve；
The quality (g) of M: sample；
V1: the volume (mL) of taken sample test solution during mensuration；
Vo: the constant volume (mL) after sample treatment.
The mensuration of the chelating rate of ferrum:
M in formula1: the amount (mg) of ferrum in chelate；
M0: add the total amount (mg) of ferrum in reaction system.
The chelation percent of the final Volvariella volvacea (Bull.Ex Franch.) Singer． protein peptide-ferrous chelate compound measuring preparation is 87.8%.
The foregoing is only presently preferred embodiments of the present invention, all impartial changes done according to scope of the present invention patent with
Modify, all should belong to the covering scope of the present invention.
Claims (9)
1. the preparation method of edible and medical fungi protein peptide-ferrous chelate compound, it is characterised in that: comprise the steps:
(1) protein extraction: utilize alkali extraction-acid precipitation or ammonium sulfate precipitation method to extract edible and medical fungi albumen；
(2) enzymolysis: use protease that the edible and medical fungi albumen extracted is carried out limited enzymolysis, prepares edible and medical fungi protein peptide and is combined
Thing solution；
(3) prepared by polypeptide-ferrous chelate compound: utilize solid high-temperature mixing method or chelate liquid legal that edible and medical fungi protein peptide is multiple
Polymer solution makes edible and medical fungi protein peptide-ferrous chelate compound；
(4) edible and medical fungi protein peptide-ferrous chelate compound is carried out lyophilization.
The preparation method of a kind of edible and medical fungi protein peptide-ferrous chelate compound the most according to claim 1, it is characterised in that:
Described edible and medical fungi includes: Lentinus Edodes, Auricularia, Pleurotus ostreatus, crab flavour mushroom, agrocyb eaegerita, Poria, Phellinus igniarius (L. ex Fr.) Quel., Volvariella volvacea (Bull.Ex Franch.) Singer．, Flammulina velutiper (Fr.) Sing, Fructus Pruni Bao
One in the edible and medical fungis such as mushroom, Tremella, Caulis Bambusae In Taeniam, Ganoderma, Grifola frondosa and Hericium erinaceus (Bull. Ex Fr.) Pers..
The preparation method of a kind of edible and medical fungi protein peptide-ferrous chelate compound the most according to claim 1, it is characterised in that:
In step (1), specifically comprising the following steps that first by edible and medical fungi micronizing, the food medicine after pulverizing of described alkali extraction-acid precipitation
Mix with the ratio of 1 g:20-60 mL with distilled water with bacterium, utilize the NaOH or KOH solution regulation pH of 0.5wt%-2wt% extremely
9.0-11.0, first supersound extraction 10-50 min, then extract 60-120 min 50-100 DEG C of water-bath, filter after extraction, take filter
Liquid, filtrate is the edible and medical fungi protein solution that alkali carries, and the edible and medical fungi protein solution pH to 2.5-4.0 carried with hydrochloric acid regulation alkali enters
Row albumen precipitation, 3000-5000 r/min is centrifuged 5-10 min and obtains protein precipitation, is washed to neutral recentrifuge, will precipitate cold
Freeze and be drying to obtain edible and medical fungi albumen.
The preparation method of a kind of edible and medical fungi protein peptide-ferrous chelate compound the most according to claim 1, it is characterised in that:
In step (1), specifically comprising the following steps that first by edible and medical fungi micronizing, the food after pulverizing of described ammonium sulfate precipitation method
Medicinal fungus mixes with the ratio of 1 g:20-60 mL with distilled water, is gradually added solid ammonium sulfate and makes the saturation of ammonium sulfate reach
40-60wt%, 3000-5000 r/min is centrifuged 5-10 min and obtains edible and medical fungi albumen precipitation, freezing dry after utilizing dialysis desalination
Dry edible and medical fungi albumen.
The preparation method of a kind of edible and medical fungi protein peptide-ferrous chelate compound the most according to claim 1, it is characterised in that:
In step (2), edible and medical fungi protein dissolution is made albumen quality concentration reach in water by specifically comprising the following steps that of described limited enzymolysis
1.0-5.0%, regulates pH to 7.0-11.0, and temperature is maintained at 40-60 DEG C, adds the protease of 1.0-5.0 wt%, hydrolyzes 30-
90 min, enzyme denaturing, obtain edible and medical fungi protein polypeptide complex solution.
The preparation method of a kind of edible and medical fungi protein peptide-ferrous chelate compound the most according to claim 1, it is characterised in that:
In step (3), edible and medical fungi protein polypeptide complex solution is dried system by specifically comprising the following steps that of described solid high-temperature mixing method
Obtain lyophilized powder；Ferrous chloride or ferrous sulfate powder and lyophilized powder are mixed according to mass ratio 1:6-5:6,300-500 DEG C of heating
1-3h prepares edible and medical fungi protein peptide-ferrous chelate compound.
The preparation method of a kind of edible and medical fungi protein peptide-ferrous chelate compound the most according to claim 1, it is characterised in that:
In step (3), what described chelate liquid was legal specifically comprises the following steps that edible and medical fungi protein polypeptide complex solution and 0.5-2
Mol/L ferrous chloride or copperas solution mixing, stir, and regulation pH is 3.0-9.0, continues under the conditions of 30-80 DEG C
Reaction 1.0-2.0 h, cools down after reaction, and the removal of impurity is gone in centrifugation, adds the 75-95%(v/v of centrifugal liquid 3-5 times volume)
Ethanol, precipitation stands 12-24 h, centrifugal segregation supernatant, it is thus achieved that edible and medical fungi protein peptide-ferrous chelate compound；Wherein, protochloride
The volume ratio of ferrum or copperas solution and edible and medical fungi protein polypeptide complex solution is 1:6-5:6.
The preparation method of a kind of edible and medical fungi protein peptide-ferrous chelate compound, its feature exists
In: described protease is alkaline protease, neutral protease or compound protease.
9. edible and medical fungi protein peptide-ferrous chelate compound that the preparation method as described in any one of claim 1-7 prepares.
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