CN112710822A - In-vitro simulated digestion method for edible fungus polysaccharide and selenizing derivative thereof - Google Patents
In-vitro simulated digestion method for edible fungus polysaccharide and selenizing derivative thereof Download PDFInfo
- Publication number
- CN112710822A CN112710822A CN202011495304.1A CN202011495304A CN112710822A CN 112710822 A CN112710822 A CN 112710822A CN 202011495304 A CN202011495304 A CN 202011495304A CN 112710822 A CN112710822 A CN 112710822A
- Authority
- CN
- China
- Prior art keywords
- selenium
- polysaccharide
- solution
- nano
- sample
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 103
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 103
- 150000004676 glycans Chemical class 0.000 title claims abstract description 101
- 230000029087 digestion Effects 0.000 title claims abstract description 57
- 238000000034 method Methods 0.000 title claims abstract description 29
- 238000000338 in vitro Methods 0.000 title claims abstract description 26
- 241000233866 Fungi Species 0.000 title claims abstract description 16
- 229910052711 selenium Inorganic materials 0.000 claims abstract description 132
- 239000011669 selenium Substances 0.000 claims abstract description 131
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 claims abstract description 109
- 229940091258 selenium supplement Drugs 0.000 claims description 122
- 239000002245 particle Substances 0.000 claims description 39
- 239000000243 solution Substances 0.000 claims description 32
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 27
- 235000000346 sugar Nutrition 0.000 claims description 26
- 239000000523 sample Substances 0.000 claims description 23
- 229920002451 polyvinyl alcohol Polymers 0.000 claims description 21
- 241000222455 Boletus Species 0.000 claims description 20
- 101710185445 Cytochrome c peroxidase, mitochondrial Proteins 0.000 claims description 20
- 238000000502 dialysis Methods 0.000 claims description 20
- 238000002360 preparation method Methods 0.000 claims description 19
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 19
- 239000012498 ultrapure water Substances 0.000 claims description 19
- 150000002772 monosaccharides Chemical class 0.000 claims description 15
- 239000000203 mixture Substances 0.000 claims description 14
- 239000000126 substance Substances 0.000 claims description 13
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims description 12
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 claims description 12
- 239000000843 powder Substances 0.000 claims description 12
- 230000001079 digestive effect Effects 0.000 claims description 11
- BVTBRVFYZUCAKH-UHFFFAOYSA-L disodium selenite Chemical compound [Na+].[Na+].[O-][Se]([O-])=O BVTBRVFYZUCAKH-UHFFFAOYSA-L 0.000 claims description 11
- 229960001471 sodium selenite Drugs 0.000 claims description 11
- 239000011781 sodium selenite Substances 0.000 claims description 11
- 235000015921 sodium selenite Nutrition 0.000 claims description 11
- 239000006228 supernatant Substances 0.000 claims description 11
- 238000006243 chemical reaction Methods 0.000 claims description 9
- 239000012530 fluid Substances 0.000 claims description 9
- 235000011389 fruit/vegetable juice Nutrition 0.000 claims description 9
- 238000002156 mixing Methods 0.000 claims description 9
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 8
- 230000003078 antioxidant effect Effects 0.000 claims description 7
- 230000002496 gastric effect Effects 0.000 claims description 7
- 230000000968 intestinal effect Effects 0.000 claims description 7
- 102000004169 proteins and genes Human genes 0.000 claims description 7
- 108090000623 proteins and genes Proteins 0.000 claims description 7
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 6
- 240000001080 Grifola frondosa Species 0.000 claims description 6
- 235000007710 Grifola frondosa Nutrition 0.000 claims description 6
- 229960005070 ascorbic acid Drugs 0.000 claims description 6
- 239000011668 ascorbic acid Substances 0.000 claims description 6
- 235000010323 ascorbic acid Nutrition 0.000 claims description 6
- 239000001110 calcium chloride Substances 0.000 claims description 6
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 6
- 239000012153 distilled water Substances 0.000 claims description 6
- 238000001035 drying Methods 0.000 claims description 6
- 238000004108 freeze drying Methods 0.000 claims description 6
- 229910001629 magnesium chloride Inorganic materials 0.000 claims description 6
- 239000011259 mixed solution Substances 0.000 claims description 6
- 238000007873 sieving Methods 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 102000004190 Enzymes Human genes 0.000 claims description 5
- 108090000790 Enzymes Proteins 0.000 claims description 5
- 229940088598 enzyme Drugs 0.000 claims description 5
- 239000000463 material Substances 0.000 claims description 5
- 239000003833 bile salt Substances 0.000 claims description 4
- 239000012488 sample solution Substances 0.000 claims description 4
- 239000011780 sodium chloride Substances 0.000 claims description 4
- 239000004382 Amylase Substances 0.000 claims description 3
- 102000013142 Amylases Human genes 0.000 claims description 3
- 108010065511 Amylases Proteins 0.000 claims description 3
- 102100031416 Gastric triacylglycerol lipase Human genes 0.000 claims description 3
- 108010019160 Pancreatin Proteins 0.000 claims description 3
- 102000057297 Pepsin A Human genes 0.000 claims description 3
- 108090000284 Pepsin A Proteins 0.000 claims description 3
- 235000019418 amylase Nutrition 0.000 claims description 3
- 238000009835 boiling Methods 0.000 claims description 3
- 238000005119 centrifugation Methods 0.000 claims description 3
- 239000000385 dialysis solution Substances 0.000 claims description 3
- 108010091264 gastric triacylglycerol lipase Proteins 0.000 claims description 3
- 229940055695 pancreatin Drugs 0.000 claims description 3
- 229940111202 pepsin Drugs 0.000 claims description 3
- 230000001502 supplementing effect Effects 0.000 claims description 3
- 241000221787 Erysiphe Species 0.000 claims description 2
- 238000002791 soaking Methods 0.000 claims description 2
- 238000011160 research Methods 0.000 abstract description 16
- 238000002474 experimental method Methods 0.000 abstract description 7
- 241001465754 Metazoa Species 0.000 abstract description 6
- 238000010521 absorption reaction Methods 0.000 abstract description 6
- 231100000419 toxicity Toxicity 0.000 abstract description 5
- 230000001988 toxicity Effects 0.000 abstract description 5
- 230000009286 beneficial effect Effects 0.000 abstract description 4
- 230000004071 biological effect Effects 0.000 description 7
- 210000001035 gastrointestinal tract Anatomy 0.000 description 6
- 229960001031 glucose Drugs 0.000 description 6
- 239000002105 nanoparticle Substances 0.000 description 6
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 5
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical class OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 5
- 210000004027 cell Anatomy 0.000 description 5
- 238000012512 characterization method Methods 0.000 description 5
- 239000008103 glucose Substances 0.000 description 5
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 4
- 230000002776 aggregation Effects 0.000 description 4
- 238000004220 aggregation Methods 0.000 description 4
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 4
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 4
- 230000007423 decrease Effects 0.000 description 4
- 238000009826 distribution Methods 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 239000007791 liquid phase Substances 0.000 description 4
- 230000002503 metabolic effect Effects 0.000 description 4
- 235000015097 nutrients Nutrition 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 210000002784 stomach Anatomy 0.000 description 4
- 239000011573 trace mineral Substances 0.000 description 4
- 235000013619 trace mineral Nutrition 0.000 description 4
- 241001489124 Boletus edulis Species 0.000 description 3
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 3
- 239000013543 active substance Substances 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 230000006378 damage Effects 0.000 description 3
- 235000014113 dietary fatty acids Nutrition 0.000 description 3
- 239000000194 fatty acid Substances 0.000 description 3
- 229930195729 fatty acid Natural products 0.000 description 3
- 150000004665 fatty acids Chemical class 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 229930182830 galactose Natural products 0.000 description 3
- 238000004817 gas chromatography Methods 0.000 description 3
- 230000004060 metabolic process Effects 0.000 description 3
- 210000000214 mouth Anatomy 0.000 description 3
- 229920000642 polymer Polymers 0.000 description 3
- 238000006722 reduction reaction Methods 0.000 description 3
- 210000000813 small intestine Anatomy 0.000 description 3
- 238000001179 sorption measurement Methods 0.000 description 3
- 150000008163 sugars Chemical class 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 241000209219 Hordeum Species 0.000 description 2
- 238000009825 accumulation Methods 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 238000000540 analysis of variance Methods 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- PYMYPHUHKUWMLA-WDCZJNDASA-N arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 0.000 description 2
- 238000004166 bioassay Methods 0.000 description 2
- 230000015556 catabolic process Effects 0.000 description 2
- 238000007405 data analysis Methods 0.000 description 2
- 210000002249 digestive system Anatomy 0.000 description 2
- 239000008151 electrolyte solution Substances 0.000 description 2
- 235000020776 essential amino acid Nutrition 0.000 description 2
- 239000003797 essential amino acid Substances 0.000 description 2
- 238000000556 factor analysis Methods 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 238000001095 inductively coupled plasma mass spectrometry Methods 0.000 description 2
- 230000004792 oxidative damage Effects 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 210000003296 saliva Anatomy 0.000 description 2
- 238000004088 simulation Methods 0.000 description 2
- 238000013112 stability test Methods 0.000 description 2
- LUEWUZLMQUOBSB-FSKGGBMCSA-N (2s,3s,4s,5s,6r)-2-[(2r,3s,4r,5r,6s)-6-[(2r,3s,4r,5s,6s)-4,5-dihydroxy-2-(hydroxymethyl)-6-[(2r,4r,5s,6r)-4,5,6-trihydroxy-2-(hydroxymethyl)oxan-3-yl]oxyoxan-3-yl]oxy-4,5-dihydroxy-2-(hydroxymethyl)oxan-3-yl]oxy-6-(hydroxymethyl)oxane-3,4,5-triol Chemical compound O[C@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@@H](O[C@@H]2[C@H](O[C@@H](OC3[C@H](O[C@@H](O)[C@@H](O)[C@H]3O)CO)[C@@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O LUEWUZLMQUOBSB-FSKGGBMCSA-N 0.000 description 1
- LWFUFLREGJMOIZ-UHFFFAOYSA-N 3,5-dinitrosalicylic acid Chemical compound OC(=O)C1=CC([N+]([O-])=O)=CC([N+]([O-])=O)=C1O LWFUFLREGJMOIZ-UHFFFAOYSA-N 0.000 description 1
- 244000247812 Amorphophallus rivieri Species 0.000 description 1
- 235000001206 Amorphophallus rivieri Nutrition 0.000 description 1
- 241000221198 Basidiomycota Species 0.000 description 1
- 241000556812 Boletus junquilleus Species 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 229920001661 Chitosan Polymers 0.000 description 1
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 description 1
- 101710088194 Dehydrogenase Proteins 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 240000008397 Ganoderma lucidum Species 0.000 description 1
- 235000001637 Ganoderma lucidum Nutrition 0.000 description 1
- 229920002581 Glucomannan Polymers 0.000 description 1
- 102000006587 Glutathione peroxidase Human genes 0.000 description 1
- 108700016172 Glutathione peroxidases Proteins 0.000 description 1
- 241000219774 Griffonia Species 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 229920002752 Konjac Polymers 0.000 description 1
- SHZGCJCMOBCMKK-JFNONXLTSA-N L-rhamnopyranose Chemical compound C[C@@H]1OC(O)[C@H](O)[C@H](O)[C@H]1O SHZGCJCMOBCMKK-JFNONXLTSA-N 0.000 description 1
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-N 0.000 description 1
- 101100505672 Podospora anserina grisea gene Proteins 0.000 description 1
- RJFAYQIBOAGBLC-BYPYZUCNSA-N Selenium-L-methionine Chemical compound C[Se]CC[C@H](N)C(O)=O RJFAYQIBOAGBLC-BYPYZUCNSA-N 0.000 description 1
- RJFAYQIBOAGBLC-UHFFFAOYSA-N Selenomethionine Natural products C[Se]CCC(N)C(O)=O RJFAYQIBOAGBLC-UHFFFAOYSA-N 0.000 description 1
- 102000004531 Selenoprotein P Human genes 0.000 description 1
- 108010042443 Selenoprotein P Proteins 0.000 description 1
- 102000008114 Selenoproteins Human genes 0.000 description 1
- 108010074686 Selenoproteins Proteins 0.000 description 1
- 241000518374 Suillellus luridus Species 0.000 description 1
- 102000013090 Thioredoxin-Disulfide Reductase Human genes 0.000 description 1
- 108010079911 Thioredoxin-disulfide reductase Proteins 0.000 description 1
- 241000223104 Trypanosoma Species 0.000 description 1
- 235000010489 acacia gum Nutrition 0.000 description 1
- 239000001785 acacia senegal l. willd gum Substances 0.000 description 1
- 230000007059 acute toxicity Effects 0.000 description 1
- 231100000403 acute toxicity Toxicity 0.000 description 1
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 229940093761 bile salts Drugs 0.000 description 1
- 201000011529 cardiovascular cancer Diseases 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000002270 dispersing agent Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 210000001339 epidermal cell Anatomy 0.000 description 1
- 210000003238 esophagus Anatomy 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 229940046240 glucomannan Drugs 0.000 description 1
- -1 glucose monosaccharide Chemical class 0.000 description 1
- 230000034659 glycolysis Effects 0.000 description 1
- 241000411851 herbal medicine Species 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 231100000086 high toxicity Toxicity 0.000 description 1
- 150000002433 hydrophilic molecules Chemical class 0.000 description 1
- 230000002218 hypoglycaemic effect Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 239000000252 konjac Substances 0.000 description 1
- 235000010485 konjac Nutrition 0.000 description 1
- 210000002429 large intestine Anatomy 0.000 description 1
- 235000004213 low-fat Nutrition 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000007102 metabolic function Effects 0.000 description 1
- 230000003278 mimic effect Effects 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 239000002366 mineral element Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000007709 nanocrystallization Methods 0.000 description 1
- 231100000956 nontoxicity Toxicity 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 231100000915 pathological change Toxicity 0.000 description 1
- 230000036285 pathological change Effects 0.000 description 1
- 210000003800 pharynx Anatomy 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 150000004804 polysaccharides Polymers 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000000529 probiotic effect Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 150000004671 saturated fatty acids Chemical class 0.000 description 1
- 229940065287 selenium compound Drugs 0.000 description 1
- 229960002718 selenomethionine Drugs 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 230000000087 stabilizing effect Effects 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 230000001960 triggered effect Effects 0.000 description 1
- 150000004670 unsaturated fatty acids Chemical class 0.000 description 1
- 235000021122 unsaturated fatty acids Nutrition 0.000 description 1
- 238000003809 water extraction Methods 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Polymers & Plastics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Materials Engineering (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Food Science & Technology (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
- Biomedical Technology (AREA)
- Sustainable Development (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses an in vitro simulated digestion method of edible fungus polysaccharide and selenizing derivatives thereof, which is characterized in that the influence of in vitro simulated digestion on polysaccharide and selenizing derivatives thereof is firstly researched, and the result shows that the polysaccharide modified by nano selenium is more beneficial to being digested and absorbed by human bodies, so that the polysaccharide can be selectively absorbed in a targeted manner, and the bioactivity of the polysaccharide can be better exerted; compared with the current research on the toxicity of inorganic selenium, organic selenium and nano selenium, the invention compares the bioacessability of selenium in the inorganic selenium, the organic selenium and the nano selenium for the first time, promotes the absorption of the selenium in the national safety limit standard range, and compared with animal experiments, the experiment is more rapid and convenient, and the operability and the repeatability are high; compared with the current research, the method is more comprehensive, and the method is suitable for all polysaccharides and selenylation derivatives thereof.
Description
Technical Field
The invention relates to the technical field of biological metabolism, in particular to an in-vitro simulated digestion method of edible fungus polysaccharide and selenizing derivatives thereof.
Background
Selenium is an essential trace element in human and animals, and is involved in a very important metabolic function in the body. On the one hand, selenium is an important component of various active selenoproteins, such as: glutathione peroxidase (GSH-Px), thioredoxin reductase, selenoprotein P and the like with antioxidant activity, on the other hand, if the organism lacks selenium element, partial enzyme activity is reduced, so that the antioxidant damage capability of tissue cells is weakened, and a series of pathological changes are directly triggered, such as: oxidative damage to cells, cardiovascular diseases, cancer, and the like. Recent research also finds that the selenium element can remarkably improve the oxidation resistance of the organism and inhibit the oxidative damage of cells, thereby maintaining the structure and the function of the cells.
Nanotechnology refers to the emerging scientific technology of manipulating atoms, molecules, or atomic groups, molecular groups on the surface of a substance on the nanoscale (0.1-100nm) to rearrange and combine them to have new functions or produce specific products. Many studies have demonstrated that food and nutrients, after nanocrystallization, exhibit higher biological activity, even those not exhibited by normal substances. Studies have shown that the bioavailability and toxicity of elemental selenium depends on the chemical state of selenium. The acute toxicity of nano-selenium is 1/7 of sodium selenite, 1/3 of organic selenium (113.0 mg/kg, 15.7 mg/kg and 30-40mg/kg of LD50 of nano-selenium, inorganic selenium and organic selenium respectively), which is considered to be the highest safety in the known selenium preparation. Nano selenium is a red colloidal state elemental selenium (zero-valent selenium), has almost no toxicity, is easier to absorb than inorganic selenium, and shows better biological activity.
However, in the liquid phase, the red nano-selenium is very unstable and easily aggregated to form gray or black selenium in the form of large-particle powder if no dispersant or stabilizer exists, thereby losing bioactivity and bioavailability. In order to prevent further aggregation of the nano-selenium particles, it is necessary to modify the surface of the nano-selenium particles and disperse the nano-selenium particles. Recent research shows that the polysaccharide has a complex branch structure and active hydroxyl groups, so that the polysaccharide can well adsorb and wrap nano-selenium initially formed in a reduction reaction in the preparation process of the nano-selenium, and nano-selenium particles can exist in a liquid phase system more stably. Research has shown that polysaccharides such as konjac glucomannan, Arabic gum, ganoderma lucidum polysaccharide and chitosan can prevent the aggregation of nano selenium and improve the stability of the nano selenium in a liquid phase. In addition, a number of studies have shown that the biological activity of the polysaccharide itself also confers a richer biological profile to the polysaccharide-nanoselenium complex.
Boletus is a rare wild edible fungus, is one of four main fungi, and is recorded in Yunnan herbal medicine. The brown yellow bolete (Suillellus luridus) is one of bolete, and is mainly distributed in broad-leaved forest areas such as Yunnan, Hebei and Guangdong. The strain is large in size, thick in meat, tender in texture and unique in volatile flavor components. The brown boletus edulis fruiting body contains rich mineral substances and reasonable amino acid and fatty acid compositions, particularly has the characteristics of high protein, low fat, low energy and the like, and researches show that the hypoglycemic activity and the antioxidant activity of the brown boletus edulis polysaccharide are obviously superior to those of other boletus edulis.
Trypanosoma griseum (Cratellus cornucopiaides) belongs to Basidiomycota, is a precious wild edible fungus and is widely distributed in Europe, North America and Asia. Because of its crisp nature and strong fruit fragrance, it has a history of eating Huimiqin fungus for a long time in China, and it is mainly distributed in Yunnan, Sichuan, Anwei, Zhejiang, Fujian, Hunan, Guangxi, Shaanxi and Tibet. Early tests show that the grifola frondosa sporocarp is rich in protein and polysaccharide and low in fat content, the content of unsaturated fatty acid in the fatty acid proportion is obviously higher than that of saturated fatty acid, and in addition, the grifola frondosa sporocarp contains a plurality of amino acids and mineral elements, and the proportion of essential amino acids and non-essential amino acids is 0.82 and is higher than 0.6 recommended by FAO/WHO (fatty acid/protein dehydrogenase), so that the grifola frondosa can be used as a good source of some nutrients, is an edible fungus with delicious taste and high nutritional value, and has high economic value and wide development prospect. Studies have shown that polysaccharides are the major active species in the hymenophora grisea sporophore.
The digestion is a complex metabolic activity in vivo, which is a key step for realizing the metabolic absorption of nutrient substances of human bodies, the digestive system consists of mouth, pharynx, esophagus, stomach, small intestine and large intestine, and the physiological efficacy of active substances is related to the digestion and glycolysis processes of the active substances in human bodies to a certain extent. Today the way food is digested and how it functions in the body has become a research hotspot. If the human body or the animal is taken as an experimental object for digestion research, factors such as moral constraints, long experimental period, large consumption, individual differences and the like need to be taken into consideration. However, the in vitro digestion model simulates the human body digestion environment to a certain extent, and has been widely used for the research of the bioavailability and the bioactivity of nutrient active substances due to the characteristics of simple operation, good repeatability, short time consumption, low cost and the like. Polysaccharides, which are high molecular hydrophilic compounds, are difficult to directly act on cells other than intestinal epidermal cells and the like in the human body. However, there are studies that indicate that pH, bile salts and enzymes in the digestive medium of the gastrointestinal tract alter the molecular weight, chemical composition, structure and conformation of polysaccharides, which are related to the biological activity of polysaccharides.
The biological activity of edible fungi polysaccharide and selenizing derivatives thereof has been widely researched, and the research on the digestion of polysaccharide and selenizing derivatives thereof is a mechanism for solving the physiological effect thereof from a new direction. However, at present, there are few studies and reports on the in vitro digestion behavior of edible fungi polysaccharide and selenized derivatives thereof at home and abroad. Therefore, the method aims to disclose the changes of the digestion performance and the antioxidant activity of the edible fungus polysaccharide and the selenizing derivative thereof by using an in-vitro digestion model, and further better illustrate the metabolic behavior and the probiotic effect of the edible fungus polysaccharide and the selenizing derivative thereof from the perspective of digestion and metabolism.
Disclosure of Invention
In order to achieve the purpose, the invention provides the following technical scheme: an in-vitro simulated digestion method for edible fungus polysaccharide and selenizing derivatives thereof is characterized by comprising the following process steps:
material pretreatment:
drying Boletus fulvus at 60-70 deg.C to constant weight, and sieving with 60 mesh sieve to obtain dry Boletus fulvus powder;
drying Erysiphelus griseus at 60-70 deg.C to constant weight, and sieving with 60 mesh sieve to obtain dry powder of Erysiphelus griseus;
② preparing polysaccharide:
a sample of Boletus flavus powder was soaked overnight in 95% ethanol to decolorize and defat, and after centrifugation (2655 Xg, 10min), the supernatant was removed. Extracting the residue with distilled water (1:40, 85 deg.C, 4h) for 2 times, concentrating the supernatant under reduced pressure, removing protein by Sevag method, and dialyzing to remove low molecular weight substances (less than or equal to 500 Da). Subsequently, adding 4 times volume of ethanol (48h, 4 ℃) into the solution, centrifuging (2655 Xg, 10min), and freeze-drying and collecting to obtain boletus flavus polysaccharides (cSLPs);
soaking Erysiphe cinerea powder sample in 95% ethanol overnight for decolorizing and defatting, centrifuging (2655 Xg, 10min), removing supernatant, extracting residue with distilled water (1:40, 85 deg.C, 4h) for 2 times, concentrating supernatant under reduced pressure, removing protein by Sevag method, and dialyzing to remove low molecular weight substances (less than or equal to 500 Da). Adding 4 times volume of ethanol (48h, 4 ℃) into the solution, centrifuging (2655 Xg, 10min), and freeze-drying to obtain the grifola frondosa polysaccharides (CCPs);
preparing polysaccharide-nano selenium:
mixing 5mL of sodium selenite solution (2mM) with the cSLPs solution (1000mg/L), adding 6mL of ultrapure water, placing on a magnetic stirrer, fully mixing uniformly (2min), slowly adding ascorbic acid on the magnetic stirrer, reacting for 2h, placing the reaction solution in a dialysis bag for treatment until no selenium and sugar are detected outside the dialysis bag, wherein the obtained sample is the prepared nano selenium particles SeNL-cSLPs;
mixing 5mL of sodium selenite solution (2mM) with CCPs solution (1000mg/L), adding 6mL of ultrapure water, placing on a magnetic stirrer, fully mixing uniformly (2min), slowly adding ascorbic acid on the magnetic stirrer, reacting for 2h, placing the reaction solution in a dialysis bag for treatment until no selenium and sugar are detected outside the dialysis bag, and obtaining a sample which is the prepared nano-selenium particles SeNL-CCPs;
preparation of simulated digestive juice
According to 30.2mL KCl, 7.4mL KH2PO4、13.6mL NaHCO3、1.0mL MgCl2·6H2O、 0.12mL(NH4)2CO3、0.18mL HCl、0.05mL CaCl2·2H2Preparing simulated oral fluid (SSF) by the proportion of O;
according to 13.8mL KCl, 1.8mL KH2PO4、25mL NaHCO3、23.6mL NaCl、 0.8mL MgCl2·6H2O、1.0mL(NH4)2CO3、2.6mL HCl、0.01mL CaCl2·2H2Preparing Simulated Gastric Fluid (SGF) by using the proportion of O;
according to 13.6mL KCl, 1.6mL KH2PO4、85mL NaHCO3、19.2mL NaCl、 2.2mL MgCl2·6H2O、1.4mL HCl、0.08mL CaCl2·2H2And preparing simulated intestinal juice (SIF) according to the proportion of O.
Further, 40mL of SSF was mixed with 50mL of a sample solution of 8mg/mL, 5mL of salivary amylase was added, pH was adjusted to 7.0, and incubation was carried out for 5min (37 ℃,150rpm) after supplementing with ultrapure water to a volume of 100 mL;
then taking the mixed solution, adding 80mL of SGF solution, 5mL of pepsin (2000U/mL) and 5mL of gastric lipase (60U/mL), adjusting the pH value to 3.0 by HCl (6mol/L), supplementing ultrapure water to the volume of 200mL, and then incubating for 2h (37 ℃,150 rpm);
and adding 80mL of SIF solution, 25mL of bile salt (10mM) and 5g of pancreatin (100U/mL) into the mixed solution, adding HCl (6mol/L) to adjust the pH value to 7.0, then adding ultrapure water to 400mL of the mixed solution, incubating for 2h (37 ℃,150rpm), boiling for 10min after each step of digestion to inactivate the enzyme, centrifuging, dialyzing (the molecular interception amount is 300Da), collecting internal and external dialyzates respectively, concentrating under reduced pressure, lyophilizing the sample in the dialysis bag, and lyophilizing the sample outside the dialysis bag to determine the molecular weight and the antioxidant activity, and lyophilizing the sample outside the dialysis bag to determine reducing sugar and free monosaccharide.
Further, ultrapure water and simulated digestive juice were used as blank control.
Compared with the prior art, the invention has the beneficial effects that:
at present, the preparation of polysaccharide-nano selenium particles mainly focuses on researching the influence of different preparation conditions on nano particles, and the invention researches the influence of different polysaccharides on the particle size, the potential and the selenium content of the nano particles in the preparation process of polysaccharide-nano selenium for the first time, thereby further perfecting the preparation of a polysaccharide-nano selenium particle system;
the influence of in vitro simulated digestion on the polysaccharide in the polysaccharide and the selenizing derivatives thereof is explored for the first time, and the result shows that the polysaccharide modified by the nano selenium is more beneficial to being digested and absorbed by a human body, so that the polysaccharide can be selectively absorbed in a targeted manner, and the biological activity of the polysaccharide can be better exerted;
compared with the current research on the toxicity of inorganic selenium, organic selenium and nano selenium, the invention compares the bioacessability of selenium in the inorganic selenium, the organic selenium and the nano selenium for the first time, promotes the absorption of the selenium in the national safety limit standard range, and compared with animal experiments, the experiment is more rapid and convenient, and the operability and the repeatability are high;
the method mainly compares the digestive metabolic characteristics of the polysaccharides in the polysaccharides and the selenylation derivatives thereof from different sources and the biological accessibility of the selenium element, and is more comprehensive compared with the current research, and the method is suitable for all the polysaccharides and the selenylation derivatives thereof;
drawings
FIG. 1 illustrates a method for preparing a nano-selenium-polysaccharide complex;
FIG. 2 is a graph of bioassays of different types of selenium;
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1:
1. preparation of Experimental materials
Drying Boletus fulvus at 60-70 deg.C to constant weight, and sieving with 60 mesh sieve to obtain dry Boletus fulvus powder;
drying Erysiphelus griseus at 60-70 deg.C to constant weight, and sieving with 60 mesh sieve to obtain dry powder of Erysiphelus griseus;
1.2 preparation of polysaccharides
The powder samples of Boletus flavus and Hordeum griseum were soaked overnight in 95% ethanol to decolorize and defat. After centrifugation (2655 Xg, 10min), the supernatant was removed. The residue was extracted with distilled water (1:40, 85 ℃, 4h) 2 times, and the supernatant was concentrated under reduced pressure. Removing proteins by Sevag method, and dialyzing to remove low molecular weight substances (less than or equal to 500 Da). Subsequently, 4 volumes of ethanol (48h, 4 ℃) were added to the solution. Centrifuging (2655 Xg, 10min), lyophilizing, and collecting to obtain Boletus flavus polysaccharides (cSLPs) and Hordeum griseum polysaccharides (CCPs). 1.3 preparation and characterization of polysaccharide-Nano selenium
1.3.1 preparation of polysaccharide-Nano selenium
5mL of sodium selenite solution (2mM) was mixed with cSLPs and CCPs solution (1000mg/L), respectively, and then 6mL of ultrapure water was added thereto and placed on a magnetic stirrer to be sufficiently mixed (2 min). And after uniformly mixing, slowly adding ascorbic acid into a magnetic stirrer, reacting for 2 hours, placing the reaction solution into a dialysis bag for treatment until no selenium and sugar are detected outside the dialysis bag, wherein the obtained sample is the prepared nano selenium particles, and is named as SeNL-cSLPs and SeNL-CCPs respectively, and is named as SeNLs by using distilled water as a blank control.
1.3.2 characterization of polysaccharide-Nano-selenium
The particle sizes and the potentials of the prepared SeNLs, SeNL-cSLPs and SeNL-CCPs complexes are characterized by using a Malvern nanometer particle size analyzer, and the selenium content in the sample is measured by ICP-MS. And (4) performing a time stability test of the polysaccharide-nano selenium particles, standing in a refrigerator at 4 ℃, and observing an aggregation phenomenon.
1.4 simulated oral, gastric, and intestinal digestion of polysaccharides and their selenized derivatives
1.4.1 simulated digestive juices
Simulated oral fluid (SSF), Simulated Gastric Fluid (SGF) and Simulated Intestinal Fluid (SIF) were prepared in advance, and the preparation protocol is shown in table 1. And determining the polysaccharide content in the cSLPs, CCPs, SeNL-cSLPs and SeNL-CCPs, performing in-vitro simulated digestion under the condition of ensuring the consistent polysaccharide content, and researching the influence of nano-selenium modified on the digestion and absorption of the polysaccharide.
TABLE 1 amount of electrolyte solution used
Table 1Amount of electrolyte solution
1.4.2 in vitro simulated digestion
40mL of SSF was mixed with 50mL of the sample solution at 8mg/mL, 5mL of salivary amylase was added, the pH was adjusted to 7.0, and the mixture was incubated for 5min (37 ℃ C., 150rpm) after adding ultrapure water to a volume of 100 mL. Then, 80mL of SGF solution, 5mL of pepsin (2000U/mL) and 5mL of gastric lipase (60U/mL) were added to the mixture, pH was adjusted to 3.0 with HCl (6mol/L), and the mixture was incubated for 2 hours (37 ℃ C., 150rpm) after adding ultrapure water to a volume of 200 mL. The mixture was then added with 80mL of SIF solution, 25mL of bile salt (10mM) and 5g of pancreatin (100U/mL), HCl (6mol/L) was added to adjust the pH to 7.0, and then ultrapure water was added thereto to a volume of 400mL and incubated for 2 hours (37 ℃ C., 150 rpm). After each digestion step, boiling for 10min to inactivate the enzyme. Centrifuging, dialyzing (molecular interception amount of 300Da), collecting internal and external dialyzates, concentrating under reduced pressure, and lyophilizing. And freeze-drying the sample in the dialysis bag for measuring molecular weight and antioxidant activity, and freeze-drying the sample outside the dialysis bag for measuring reducing sugar and free monosaccharide. In addition, the experiment was performed with the blank of ultrapure water plus simulated digestive fluid, and the blank was subtracted from all the test results.
1.5 polysaccharide and derivatives thereof mimic post-digestion changes
1.5.1 determination of reducing sugar content
Taking 1.4.2 supernatant, determining the content of reducing sugar by a 3, 5-dinitrosalicylic acid (DNS) method, and constructing a standard curve by a series of anhydrous glucose standard products.
1.5.2 determination of free monosaccharides
A freeze-dried sample outside the 1.4.2 dialysis bag was taken, and the sample was acetylated and then free monosaccharide was measured by Gas Chromatography (GC).
1.5.3 determination of the molecular weight distribution
The lyophilized sample in the 1.4.2 dialysis bag was reconstituted and passed through a 0.22 μm aqueous membrane, and the molecular weight of ORPS was determined by High Performance Gas Permeation Chromatography (HPGPC) using an Agilent 1100 high performance liquid chromatography system, column temperature: the mixture was eluted at 30 ℃ with ultrapure water at a flow rate of 0.8 mL/min. A sample of polysaccharide sample solution was injected at 20. mu.L. A series of dextran standards were prepared to prepare a standard curve.
1.6 data analysis processing
All data are presented as mean ± standard deviation, single-factor analysis of variance (ANOVA) using SPSS 22.0 software, and Tukey's multiple comparisons, with differences statistically significant at p < 0.05.
2 results and analysis
2.1 preparation of polysaccharides
The boletus flavus polysaccharide cSLPs are obtained by a water extraction and alcohol precipitation method, and then the structure of the boletus flavus polysaccharide cSLPs is preliminarily characterized. From the GC results (table 2), it can be shown that the cSLPs consist of arabinose (10.67%), xylose (2.06%), mannose (19.66%), glucose (46.07%) and galactose (21.53%), with glucose and galactose predominating. In addition, the HPGPC chromatogram of the cSLPs had 1 peak, indicating that the molecular weight was predominantly 6383 Da.
The griffonia polysaccharide CCPs are mainly composed of rhamnose (0.55%), arabinose (0.33%), xylose (18.07%), mannose (42.25%), glucose (13.02%) and galactose (16.55%), wherein mannose is dominant. In addition, the HPGPC chromatogram of CCPs had two peaks, indicating that the molecular weights were mainly distributed between 518,019 and 14,579Da, in proportions of 44.86% and 55.14%, respectively, and the average molecular weight was 240.422 Da. The proportions of the monosaccharide compositions and the molecular weight distributions are different from those in the previous researches, but the proportions and the molecular weight distributions are reasonable in the research of the polysaccharide, because the polysaccharide structure is influenced by factors such as the source of the polysaccharide, the extraction method, the environment, the temperature and the like.
TABLE 2 molecular weights and monosaccharide compositions of cSLPs and CCPs
Table 2Molecular weight(Da)and monosaccharides composition(％)of cSLPs and CCPs.
2.2 characterization of polysaccharide-Nano-selenium
The reaction principle of sodium selenite and ascorbic acid in a polysaccharide aqueous solution is shown in figure 1, the polysaccharide has a complex branch structure and active hydroxyl groups, so that the polysaccharide is added in the preparation process of nano-selenium, nano-selenium initially formed in the reduction reaction can be well adsorbed and coated, nano-selenium particles are enabled to exist in a liquid phase system more stably, and the polysaccharide self-bioactivity endows the polysaccharide-nano-selenium complex with richer biological characteristics. During the reaction, the solution of the reaction system gradually changes from colorless to red, which shows that polysaccharide-nano selenium particles are generated. The particle size of the polysaccharide nano-selenium prepared from the boletus flavus polysaccharide, the grifola frondosa polysaccharide and the sodium selenite is shown in table 3, and the particle size of the two polysaccharides, namely nano-selenium particles, is obviously smaller than that of sugar-free nano-selenium, so that the polysaccharides have better physical adsorption effect, and are wrapped with the nano-selenium particles in the system to form a polysaccharide-nano-selenium compound. The particle size of the boletus flavus polysaccharide-nano selenium particles (SeNL-cSLPs) is smaller than that of the trumpetus griseus polysaccharide-nano selenium particles (SeNL-CCPs), which probably is that the particle size of the obtained nanoparticles is smaller under the same preparation conditions because the molecular weight of the boletus flavus polysaccharide is smaller, the molecular chain length of the boletus flavetus polysaccharide is shorter, the molecular chain flexibility is better, and the steric hindrance is smaller.
In addition, research results show that the embedding effect of polysaccharides with different structures on selenium particles is different, although the selenium content in two polysaccharides, namely nano selenium particles, is remarkably higher than that of sugar-free nano selenium, SeNL-cSLPs can better reserve the selenium content, and the selenium content is possibly smaller than that of boletus flavus polysaccharide, and the chain length is shorter, so that the selenium particle has more active-OH and higher specific surface area, and has a certain adsorption effect on elements such as selenium. Meanwhile, polysaccharide-nano selenium stability tests are carried out, although the selenium content in the SeNLs is the highest, the stability is the worst, aggregation occurs within 48 hours, and therefore, polysaccharide is very necessary for stabilizing nano selenium particles. The selenium atom in sodium selenite can associate with the polysaccharide hydroxyl groups through intermolecular hydrogen bonding. In the preparation process of the polysaccharide nano selenium, selenium in a +4 valence state in sodium selenite is reduced into selenium element in a 0 valence state, the element is aggregated to form particles, the particles grow along with the reaction, and finally, spherical nano selenium particles are formed through the combination of intermolecular hydrogen bonds and polysaccharide hydroxyl groups and the adsorption of the polysaccharide.
TABLE 3 characterization of the nanoparticles
Table 3Characterization of Nanoparticles
2.3 in vitro molecular weight changes following simulated digestion of polysaccharides and derivatives thereof
Polysaccharides from food should be available to the body through the digestive system to exert their biological activity. Thus, in this study, we have developed an in vitro simulation model to study the digestion characteristics of cSLPs, CCPs, SeNL-cSLPs and SeNL-CCPs. As shown in table 4, the Mw of the samples varied at different stages during the simulated digestion. During digestion, the molecular weight of the cSLPs gradually decreases and changes from single peak to double peak; whereas the Mw of CCP had little change in saliva digestion (from 14,579Da, 518,019Da to 12,598Da, 495,272 Da), indicating that CCP was not affected by saliva digestion. The Mw of CCPs was then observed to decrease continuously in gastrointestinal tract digestion. Compared with selenized polysaccharide, the molecular weight change of the polysaccharide-nano selenium particles is higher than that of the original sugar, so the selenized and modified polysaccharide is more easily influenced by gastrointestinal digestive juice. However, polysaccharides tend to form aggregates in solution, and water-soluble polysaccharides with similar chemical structures exhibit different chain conformations in solution. It is difficult to judge whether the decrease in Mw of the polymer is due to the destruction of aggregates or the breaking of chemical bonds in the polymer. Therefore, we further analyzed the reducing sugar content and the free monosaccharide content.
2.4 in vitro simulation of reducing sugar content after digestion of polysaccharides and derivatives thereof
If the polysaccharide is digestible, the glycosidic bond is broken, resulting in an increase in the reducing end of the sugar chain. As shown in table 4, during digestion, no significant loss of polysaccharide (p >0.05) was observed with the cSLPs and CCPs during salivary or gastric digestion, with a significant increase only after intestinal digestion, indicating that polysaccharide molecular linkages have changed and that certain glycosidic linkages have been broken. However, even after 4 hours of digestion in the simulated gastrointestinal tract, only small amounts of reducing sugars are produced, which means that only part of the glycosidic bonds are changed. The significant reduction in molecular weight may be due to the breakdown of aggregates in the polymer chain. However, the reducing sugar release amount of the polysaccharide modified by the nano-selenium is obviously higher than that of the original sugar at each stage, so the selenized polysaccharide is probably more beneficial to the digestion and absorption of the human body.
2.5 free monosaccharide composition after in vitro simulated digestion of polysaccharides and derivatives thereof
As can be seen from table 4, after simulated digestion, all samples released different degrees of free monosaccharides, and the polysaccharide-nanoselenium particles obtained the free monosaccharide distribution was closer to the composition of the raw sugar monosaccharides than the raw sugar, i.e. the polysaccharide-nanoselenium particles more favoured the release in the gastrointestinal tract. The decrease in Mw of polysaccharides and their selenized derivatives, combined with the reducing sugar content of the digestive juices, may be due to the breakdown of certain glycosidic bonds. However, the content of reducing sugars in the small intestine is still significantly lower than the total amount of polysaccharides used during digestion (4 mg/mL). Thus, despite the release of a certain amount of glucose from the digestive juices, polysaccharides and their selenized derivatives are still resistant to digestion by humans.
TABLE 4 variation of reducing sugars, monosaccharide composition, molecular weight of polysaccharides and their selenized derivatives after in vitro simulated digestion
3 conclusion
In the experiment, the in-vitro digestion model is utilized to explore the in-vitro simulated digestion metabolism characteristics of the cSLPs, the CCPs, the SeNL-cSLPs and the SeNL-CCPs, and the main conclusion is as follows:
(1) after simulated digestion in vitro of cSLPs and CCPs, the molecular weight is reduced, the reducing sugar content in the digestion solution is slightly increased, only a small amount of free glucose monosaccharide is released, which indicates that polysaccharide is possible to be depolymerized and a small amount of glycosidic bond is damaged, but the molecular structure is not completely changed
(2) After SeNL-cSLPs and SeNL-CCPs are subjected to in vitro simulated digestion, the molecular weight change, the release amount of reducing sugar and the release amount of free monosaccharide are all higher than those of original sugar, and the nano selenium modified polysaccharide is more favorable for being digested and absorbed by human bodies
Example 2:
1. experimental Material
Same as example 1, experimental material 1.
1.2 preparation of polysaccharides
The same procedure as in example 1 was repeated to prepare 1.2 polysaccharides.
1.3 preparation and characterization of polysaccharide-Nano selenium
The same procedure as in example 1 was repeated to prepare and characterize 1.3 polysaccharide-nano-selenium.
1.4 simulated oral, gastric, and intestinal digestion of polysaccharide-nano selenium particles
The specific experimental operation is the same as that of the 1.4 polysaccharide and the selenized derivatives thereof in example 1 for simulating the digestion of oral cavity, stomach and small intestine, the in vitro simulated digestion is carried out under the condition of ensuring that the selenium content of sodium selenite (inorganic selenium), selenomethionine (organic selenium), SeNL (nano-selenium), SeNL-cSLPs (nano-selenium) and SeNL-CCPs (nano-selenium) is consistent, and the bioavailabilities of different forms of selenium are researched. In addition, the experiment was performed with the blank of ultrapure water plus simulated digestive fluid, and the blank was subtracted from all the test results.
1.5 biological accessibility of elemental selenium
The selenium content of the samples was determined by ICP-MS and the selenium and bioassays calculated according to the following formula.
Biological accessibility (%) of selenium/selenium content 100% in the sample
1.6 data analysis processing
All data are presented as mean ± standard deviation, single-factor analysis of variance (ANOVA) using SPSS 22.0 software, and Tukey's multiple comparisons, with differences statistically significant at p < 0.05.
2 results and analysis
2.1 biological accessibility of elemental selenium
As shown in figure 2, the bioacessability of different types of selenium elements in the digestion process of oral cavity, stomach and intestinal tract is compared, and the results show that the bioacessability of inorganic selenium is obviously lower than that of organic selenium and nano selenium, the bioacessability of nano selenium is related to nano particles, and the bioacessability of polysaccharide-coated nano selenium particles is lower than that of sugar-free nano selenium, which is probably because the damage degree of polysaccharide in the digestion process is lower, and the release amount of reducing sugar in simulated digestion of polysaccharide in cSLPs and SeNL-cSLPs in vitro is higher than that of CCPs and SeNL-CCPs, so the bioacessability of selenium element is higher than that of CCPs and selenizing derivatives SeNL-CCPs. In addition, selenium element in organic selenium and nano selenium is mainly absorbed and accumulated from the stomach, although selenium is an indispensable trace element for human and animals, the accumulation of selenium needs to be avoided in the eating process, and SeNL has poor stability and is easy to aggregate in the digestive tract, so that the toxicity is higher.
3 conclusion
Selenium is an indispensable trace element for human and animals, but inorganic selenium has high toxicity and low absorption rate, so the biological accessibility of different types of selenium elements is compared through an in vitro digestion model, wherein the biological accessibility of nano selenium is higher than that of inorganic selenium and organic selenium, and in consideration of the accumulation of selenium, nano selenium particles with good stability are required to be selected to supplement trace elements within a safe toxicity range. Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (3)
1. An in-vitro simulated digestion method for edible fungus polysaccharide and selenizing derivatives thereof is characterized by comprising the following process steps:
material pretreatment:
drying Boletus fulvus at 60-70 deg.C to constant weight, and sieving with 60 mesh sieve to obtain dry Boletus fulvus powder;
drying Erysiphelus griseus at 60-70 deg.C to constant weight, and sieving with 60 mesh sieve to obtain dry powder of Erysiphelus griseus;
② preparing polysaccharide:
a sample of Boletus flavus powder was soaked overnight in 95% ethanol to decolorize and defat, and after centrifugation (2655 Xg, 10min), the supernatant was removed. Extracting the residue with distilled water (1:40, 85 deg.C, 4h) for 2 times, concentrating the supernatant under reduced pressure, removing protein by Sevag method, and dialyzing to remove low molecular weight substances (less than or equal to 500 Da). Subsequently, adding 4 times volume of ethanol (48h, 4 ℃) into the solution, centrifuging (2655 Xg, 10min), and freeze-drying and collecting to obtain boletus flavus polysaccharides (cSLPs);
soaking Erysiphe cinerea powder sample in 95% ethanol overnight for decolorizing and defatting, centrifuging (2655 Xg, 10min), removing supernatant, extracting residue with distilled water (1:40, 85 deg.C, 4h) for 2 times, concentrating supernatant under reduced pressure, removing protein by Sevag method, and dialyzing to remove low molecular weight substances (less than or equal to 500 Da). Adding 4 times volume of ethanol (48h, 4 ℃) into the solution, centrifuging (2655 Xg, 10min), and freeze-drying to obtain the grifola frondosa polysaccharides (CCPs);
preparing polysaccharide-nano selenium:
mixing 5mL of sodium selenite solution (2mM) with the cSLPs solution (1000mg/L), adding 6mL of ultrapure water, placing on a magnetic stirrer, fully mixing uniformly (2min), slowly adding ascorbic acid on the magnetic stirrer, reacting for 2h, placing the reaction solution in a dialysis bag for treatment until no selenium and sugar are detected outside the dialysis bag, wherein the obtained sample is the prepared nano selenium particles SeNL-cSLPs;
mixing 5mL of sodium selenite solution (2mM) with CCPs solution (1000mg/L), adding 6mL of ultrapure water, placing on a magnetic stirrer, fully mixing uniformly (2min), slowly adding ascorbic acid on the magnetic stirrer, reacting for 2h, placing the reaction solution in a dialysis bag for treatment until no selenium and sugar are detected outside the dialysis bag, and obtaining a sample which is the prepared nano-selenium particles SeNL-CCPs;
preparation of simulated digestive juice
According to 30.2mL KCl, 7.4mL KH2PO4、13.6mL NaHCO3、1.0mL MgCl2·6H2O、0.12mL (NH4)2CO3、0.18mL HCl、0.05mL CaCl2·2H2Preparing simulated oral fluid (SSF) by the proportion of O;
according to 13.8mL KCl, 1.8mL KH2PO4、25mL NaHCO3、23.6mL NaCl、0.8mL MgCl2·6H2O、1.0mL (NH4)2CO3、2.6mL HCl、0.01mL CaCl2·2H2Preparing Simulated Gastric Fluid (SGF) by using the proportion of O;
according to 13.6mL KCl, 1.6mL KH2PO4、85mL NaHCO3、19.2mL NaCl、2.2mL MgCl2·6H2O、1.4mL HCl、0.08mL CaCl2·2H2And preparing simulated intestinal juice (SIF) according to the proportion of O.
2. The method of claim 1, wherein 40mL SSF is mixed with 50mL, 8mg/mL sample solution, 5mL salivary amylase is added, pH is adjusted to 7.0, and the mixture is incubated for 5min (37 ℃,150rpm) after adding ultrapure water to a volume of 100 mL;
then taking the mixed solution, adding 80mL of SGF solution, 5mL of pepsin (2000U/mL) and 5mL of gastric lipase (60U/mL), adjusting the pH value to 3.0 by HCl (6mol/L), supplementing ultrapure water to the volume of 200mL, and then incubating for 2h (37 ℃,150 rpm);
and adding 80mL of SIF solution, 25mL of bile salt (10mM) and 5g of pancreatin (100U/mL) into the mixed solution, adding HCl (6mol/L) to adjust the pH value to 7.0, then adding ultrapure water to 400mL of the mixed solution, incubating for 2h (37 ℃,150rpm), boiling for 10min after each step of digestion to inactivate the enzyme, centrifuging, dialyzing (the molecular interception amount is 300Da), collecting internal and external dialyzates respectively, concentrating under reduced pressure, lyophilizing the sample in the dialysis bag, and lyophilizing the sample outside the dialysis bag to determine the molecular weight and the antioxidant activity, and lyophilizing the sample outside the dialysis bag to determine reducing sugar and free monosaccharide.
3. The in vitro simulated digestion method of edible fungus polysaccharide and selenizing derivatives thereof as claimed in claim 1, wherein ultrapure water and simulated digestion solution are used as blank control.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011495304.1A CN112710822A (en) | 2020-12-17 | 2020-12-17 | In-vitro simulated digestion method for edible fungus polysaccharide and selenizing derivative thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011495304.1A CN112710822A (en) | 2020-12-17 | 2020-12-17 | In-vitro simulated digestion method for edible fungus polysaccharide and selenizing derivative thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112710822A true CN112710822A (en) | 2021-04-27 |
Family
ID=75544008
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011495304.1A Pending CN112710822A (en) | 2020-12-17 | 2020-12-17 | In-vitro simulated digestion method for edible fungus polysaccharide and selenizing derivative thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112710822A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113197258A (en) * | 2021-05-14 | 2021-08-03 | 滁州市恩典硒科技咨询有限公司 | Selenium-rich vine tea and preparation method thereof |
| CN114052242A (en) * | 2021-10-27 | 2022-02-18 | 四川农业大学 | Construction of edible fungus 'ternary composite enzymolysis' and its in vitro simulated digestion method |
| CN114155770A (en) * | 2021-11-18 | 2022-03-08 | 浙江工商大学 | Method for simulating old people to digest semi-solid food based on semi-dynamic digestion system |
| CN118033100A (en) * | 2024-03-11 | 2024-05-14 | 无锡市食品安全检验检测中心 | Combined in vitro prediction method for glycemic index of liquid special medical formula food |
Citations (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040047954A1 (en) * | 2002-08-29 | 2004-03-11 | Johns Paul W. | Analytical method for the determination of infant formula protein digestibility in vitro |
| US20110136745A1 (en) * | 2008-01-03 | 2011-06-09 | Monsanto Technology Llc. | Method of selecting soybeans with enhanced bioactivity and compositions for reducing cancer cell viability |
| US20120202062A1 (en) * | 2007-12-01 | 2012-08-09 | Xueyun Gao | Selenium nanoparticles with improved biological effects |
| CN102643361A (en) * | 2012-04-25 | 2012-08-22 | 山西大学 | Nano selenium micromolecular microbial polysaccharide as well as preparation method and application thereof |
| CN103130909A (en) * | 2013-03-17 | 2013-06-05 | 吉林大学 | Preparation method of selenium-rich Morchella polysaccharide |
| US20140158021A1 (en) * | 2012-12-11 | 2014-06-12 | Wei Pan | Electrochemical Synthesis of Selenium Nanoparticles |
| CN104800240A (en) * | 2015-04-09 | 2015-07-29 | 四川农业大学 | Novel preparation method of nano selenium-catathelasma ventricosum polysaccharide complex |
| US20170191083A1 (en) * | 2016-01-04 | 2017-07-06 | Zhejiang University | Method of synthesizing biogenic elemental selenium nanostructure using enterobacter cloacae and application thereof |
| CN107760611A (en) * | 2017-10-19 | 2018-03-06 | 四川农业大学 | The liquid fermentation process optimization method and technique of a kind of selenium-rich Boletus luridus |
| CN109303922A (en) * | 2018-11-19 | 2019-02-05 | 华南理工大学 | A kind of Polysaccharides from Rosa roxburghii functionalized nano selenium compound and preparation method thereof and the application in hypoglycemic medicine |
| CN109650349A (en) * | 2018-12-29 | 2019-04-19 | 华南农业大学 | The nanometer selenium for preparing the method for nanometer selenium using camellia plant polyose and being prepared |
| CN110078840A (en) * | 2019-04-04 | 2019-08-02 | 珠海中美普莱健康科技有限公司 | A kind of Sargassum polysaccharide selenium and the preparation method and application thereof |
| CN110200287A (en) * | 2019-06-29 | 2019-09-06 | 华南理工大学 | A kind of oyster polypeptide-nano granules of selenium compound and the preparation method and application thereof with relieving alcoholism and protecting liver effect |
| CN110511297A (en) * | 2019-10-14 | 2019-11-29 | 武汉华联科生物技术有限公司 | A kind of extraction separation and purification method of selenium lentinan |
-
2020
- 2020-12-17 CN CN202011495304.1A patent/CN112710822A/en active Pending
Patent Citations (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040047954A1 (en) * | 2002-08-29 | 2004-03-11 | Johns Paul W. | Analytical method for the determination of infant formula protein digestibility in vitro |
| US20120202062A1 (en) * | 2007-12-01 | 2012-08-09 | Xueyun Gao | Selenium nanoparticles with improved biological effects |
| US20110136745A1 (en) * | 2008-01-03 | 2011-06-09 | Monsanto Technology Llc. | Method of selecting soybeans with enhanced bioactivity and compositions for reducing cancer cell viability |
| CN102643361A (en) * | 2012-04-25 | 2012-08-22 | 山西大学 | Nano selenium micromolecular microbial polysaccharide as well as preparation method and application thereof |
| US20140158021A1 (en) * | 2012-12-11 | 2014-06-12 | Wei Pan | Electrochemical Synthesis of Selenium Nanoparticles |
| CN103130909A (en) * | 2013-03-17 | 2013-06-05 | 吉林大学 | Preparation method of selenium-rich Morchella polysaccharide |
| CN104800240A (en) * | 2015-04-09 | 2015-07-29 | 四川农业大学 | Novel preparation method of nano selenium-catathelasma ventricosum polysaccharide complex |
| US20170191083A1 (en) * | 2016-01-04 | 2017-07-06 | Zhejiang University | Method of synthesizing biogenic elemental selenium nanostructure using enterobacter cloacae and application thereof |
| CN107760611A (en) * | 2017-10-19 | 2018-03-06 | 四川农业大学 | The liquid fermentation process optimization method and technique of a kind of selenium-rich Boletus luridus |
| CN109303922A (en) * | 2018-11-19 | 2019-02-05 | 华南理工大学 | A kind of Polysaccharides from Rosa roxburghii functionalized nano selenium compound and preparation method thereof and the application in hypoglycemic medicine |
| CN109650349A (en) * | 2018-12-29 | 2019-04-19 | 华南农业大学 | The nanometer selenium for preparing the method for nanometer selenium using camellia plant polyose and being prepared |
| CN110078840A (en) * | 2019-04-04 | 2019-08-02 | 珠海中美普莱健康科技有限公司 | A kind of Sargassum polysaccharide selenium and the preparation method and application thereof |
| CN110200287A (en) * | 2019-06-29 | 2019-09-06 | 华南理工大学 | A kind of oyster polypeptide-nano granules of selenium compound and the preparation method and application thereof with relieving alcoholism and protecting liver effect |
| CN110511297A (en) * | 2019-10-14 | 2019-11-29 | 武汉华联科生物技术有限公司 | A kind of extraction separation and purification method of selenium lentinan |
Non-Patent Citations (5)
| Title |
|---|
| JIXIAN ZHANG 等: "Digestion and absorption properties of Lycium barbarum polysaccharides stabilized selenium nanoparticles", 《FOOD CHEMISTRY》, vol. 373, pages 131637 * |
| YUNTAO LIU 等: "Synthesis and antidiabetic activity of selenium nanoparticles in the presence of polysaccharides from Catathelasma ventricosum", 《INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES》, vol. 114, pages 632 - 639, XP085398468, DOI: 10.1016/j.ijbiomac.2018.03.161 * |
| 曾思琪: "两种多糖-纳米硒的制备及其体内抗糖尿病活性的研究", 《中国优秀硕士学位论文全文数据库工程科技I辑》, no. 12, pages 020 - 149 * |
| 陈春: "桑葚多糖的结构鉴定、活性评价及其体外消化酵解", 《中国博士学位论文全文数据库工程科技I辑》, no. 12, pages 024 - 55 * |
| 高珍珍;张超;景丽荣;胡元亮;: "7种硒化多糖抗氧化活性的比较", 畜牧与兽医, no. 03, pages 118 - 125 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113197258A (en) * | 2021-05-14 | 2021-08-03 | 滁州市恩典硒科技咨询有限公司 | Selenium-rich vine tea and preparation method thereof |
| CN113197258B (en) * | 2021-05-14 | 2023-10-31 | 滁州市恩典硒科技咨询有限公司 | Selenium-rich vine tea and preparation method thereof |
| CN114052242A (en) * | 2021-10-27 | 2022-02-18 | 四川农业大学 | Construction of edible fungus 'ternary composite enzymolysis' and its in vitro simulated digestion method |
| CN114155770A (en) * | 2021-11-18 | 2022-03-08 | 浙江工商大学 | Method for simulating old people to digest semi-solid food based on semi-dynamic digestion system |
| CN118033100A (en) * | 2024-03-11 | 2024-05-14 | 无锡市食品安全检验检测中心 | Combined in vitro prediction method for glycemic index of liquid special medical formula food |
| CN118033100B (en) * | 2024-03-11 | 2024-07-16 | 无锡市食品安全检验检测中心 | Combined in vitro prediction method for glycemic index of liquid special medical formula food |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112710822A (en) | In-vitro simulated digestion method for edible fungus polysaccharide and selenizing derivative thereof | |
| Xie et al. | Structure, function and food applications of carboxymethylated polysaccharides: A comprehensive review | |
| Khaskheli et al. | Characterization of Auricularia auricula polysaccharides and its antioxidant properties in fresh and pickled product | |
| KR100674672B1 (en) | Chitosan-containing polysaccharide, process for producing the same and use thereof | |
| JP5094981B2 (en) | Composition of plant carbohydrates as a dietary supplement | |
| Huang et al. | Extraction, purification, structural characterization, and gut microbiota relationship of polysaccharides: A review | |
| Míguez et al. | Pectic oligosaccharides and other emerging prebiotics | |
| EP0692252A1 (en) | Composition containing inulin or oligofructose for use in cancer treatment | |
| CN102763817A (en) | Method for preparing black fungus puffed ultramicro powder | |
| Anane et al. | Improved curdlan production with discarded bottom parts of Asparagus spear | |
| Cao et al. | Positive effects of dietary fiber from sweet potato [Ipomoea batatas (L.) Lam.] peels by different extraction methods on human fecal microbiota in vitro fermentation | |
| CN113698502A (en) | Bletilla striata fibrous root oligosaccharide and preparation method and application thereof | |
| Park et al. | Other microbial polysaccharides: Pullulan, scleroglucan, elsinan, levan, alternant, dextran | |
| Akhtar et al. | Digestion by saliva, simulated gastric and small intestinal juices and in vitro fermentation by human gut microbiota of polysaccharides from cicer arietinum L. hulls: Chickpea hull polysaccharide effect on human gut | |
| CN115944706A (en) | Traditional Chinese medicine probiotic compound with blood sugar reducing function and preparation method thereof | |
| CA3026138C (en) | An edible composition for reducing the digestion or absorption of the harmful/toxic substance | |
| CN114053302A (en) | Preparation of compound edible fungus polysaccharide compound with anti-fatigue effect | |
| JPWO2003048212A1 (en) | Method for physicochemical production of glycogen and glycogen obtained by this method | |
| Varichanan et al. | Potential Prebiotic Properties of Crude Polysaccharide Extract from Durian (Durio zibethinus Murr.) Seed Flour | |
| Wang et al. | Extraction, purification, structural characteristic, health benefit, and product application of the polysaccharides from bamboo shoot: A review | |
| Furda | Chitin and chitosan–a special class of dietary fiber | |
| CN105255980A (en) | Method for producing oligosaccharide-polypeptide mixture with wheat bran | |
| Xiong et al. | Effect of Polygonatum cyrtonema Hua polysaccharides on gluten structure, in vitro digestion and shelf-life of fresh wet noodle | |
| KR100539662B1 (en) | Rice coated with a composition comprising alginic acid and porphyran | |
| Chen et al. | Evaluation of Physicochemical Properties and Prebiotics Function of a Bioactive Pleurotus eryngii Aqueous Extract Powder Obtained by Spray Drying |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20210427 |