CN103897998B - Lactobacillus salivarius and application and functional food composition and preparation method thereof - Google Patents
Lactobacillus salivarius and application and functional food composition and preparation method thereof Download PDFInfo
- Publication number
- CN103897998B CN103897998B CN201210572842.5A CN201210572842A CN103897998B CN 103897998 B CN103897998 B CN 103897998B CN 201210572842 A CN201210572842 A CN 201210572842A CN 103897998 B CN103897998 B CN 103897998B
- Authority
- CN
- China
- Prior art keywords
- lactobacillus salivarius
- food composition
- functional food
- nematicide
- present
- 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.)
- Active
Links
- 241000186869 Lactobacillus salivarius Species 0.000 title claims abstract description 121
- 239000000203 mixture Substances 0.000 title claims abstract description 47
- 235000013376 functional food Nutrition 0.000 title claims abstract description 40
- 238000002360 preparation method Methods 0.000 title abstract description 5
- 230000032683 aging Effects 0.000 claims abstract description 46
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 claims abstract description 45
- 241000894006 Bacteria Species 0.000 claims abstract description 42
- 239000000463 material Substances 0.000 claims abstract description 29
- 230000002779 inactivation Effects 0.000 claims abstract description 26
- 238000000034 method Methods 0.000 claims description 38
- 235000013305 food Nutrition 0.000 claims description 9
- 230000000694 effects Effects 0.000 abstract description 90
- 239000005645 nematicide Substances 0.000 description 62
- 210000004556 brain Anatomy 0.000 description 57
- 241000699670 Mus sp. Species 0.000 description 23
- 102000010909 Monoamine Oxidase Human genes 0.000 description 22
- 108010062431 Monoamine oxidase Proteins 0.000 description 22
- 239000012531 culture fluid Substances 0.000 description 22
- 241001167795 Escherichia coli OP50 Species 0.000 description 20
- 238000012360 testing method Methods 0.000 description 19
- 230000000052 comparative effect Effects 0.000 description 18
- 239000003651 drinking water Substances 0.000 description 17
- 235000020188 drinking water Nutrition 0.000 description 17
- 239000006228 supernatant Substances 0.000 description 17
- 239000001963 growth medium Substances 0.000 description 16
- 239000000872 buffer Substances 0.000 description 13
- 102000004190 Enzymes Human genes 0.000 description 12
- 108090000790 Enzymes Proteins 0.000 description 12
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 12
- 102000004169 proteins and genes Human genes 0.000 description 9
- 108090000623 proteins and genes Proteins 0.000 description 9
- 239000012530 fluid Substances 0.000 description 8
- 239000002609 medium Substances 0.000 description 8
- 238000001816 cooling Methods 0.000 description 7
- 239000006166 lysate Substances 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- WQZGKKKJIJFFOK-SVZMEOIVSA-N (+)-Galactose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-SVZMEOIVSA-N 0.000 description 6
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- 102000002322 Egg Proteins Human genes 0.000 description 6
- 108010000912 Egg Proteins Proteins 0.000 description 6
- 230000003712 anti-aging effect Effects 0.000 description 6
- 230000001580 bacterial effect Effects 0.000 description 6
- 230000034994 death Effects 0.000 description 6
- 230000003203 everyday effect Effects 0.000 description 6
- 239000002054 inoculum Substances 0.000 description 6
- 239000007788 liquid Substances 0.000 description 6
- 244000005700 microbiome Species 0.000 description 6
- 210000004681 ovum Anatomy 0.000 description 6
- 229910052697 platinum Inorganic materials 0.000 description 6
- 238000007789 sealing Methods 0.000 description 6
- 239000010902 straw Substances 0.000 description 6
- YVNQAIFQFWTPLQ-UHFFFAOYSA-O [4-[[4-(4-ethoxyanilino)phenyl]-[4-[ethyl-[(3-sulfophenyl)methyl]amino]-2-methylphenyl]methylidene]-3-methylcyclohexa-2,5-dien-1-ylidene]-ethyl-[(3-sulfophenyl)methyl]azanium Chemical compound C1=CC(OCC)=CC=C1NC1=CC=C(C(=C2C(=CC(C=C2)=[N+](CC)CC=2C=C(C=CC=2)S(O)(=O)=O)C)C=2C(=CC(=CC=2)N(CC)CC=2C=C(C=CC=2)S(O)(=O)=O)C)C=C1 YVNQAIFQFWTPLQ-UHFFFAOYSA-O 0.000 description 5
- 230000008859 change Effects 0.000 description 5
- 239000012153 distilled water Substances 0.000 description 5
- 238000002347 injection Methods 0.000 description 5
- 239000007924 injection Substances 0.000 description 5
- 238000002156 mixing Methods 0.000 description 5
- 238000011160 research Methods 0.000 description 5
- 230000001954 sterilising effect Effects 0.000 description 5
- 241000186660 Lactobacillus Species 0.000 description 4
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 4
- 230000009286 beneficial effect Effects 0.000 description 4
- 230000002490 cerebral effect Effects 0.000 description 4
- 238000011081 inoculation Methods 0.000 description 4
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 4
- 230000003287 optical effect Effects 0.000 description 4
- 229910052698 phosphorus Inorganic materials 0.000 description 4
- 239000011574 phosphorus Substances 0.000 description 4
- 239000006041 probiotic Substances 0.000 description 4
- 230000000529 probiotic effect Effects 0.000 description 4
- 235000018291 probiotics Nutrition 0.000 description 4
- 239000002028 Biomass Substances 0.000 description 3
- 208000027877 Disorders of Sex Development Diseases 0.000 description 3
- 229910019142 PO4 Inorganic materials 0.000 description 3
- 238000005119 centrifugation Methods 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 239000011248 coating agent Substances 0.000 description 3
- 238000000576 coating method Methods 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 230000012447 hatching Effects 0.000 description 3
- 201000005611 hermaphroditism Diseases 0.000 description 3
- 230000010196 hermaphroditism Effects 0.000 description 3
- 229940039696 lactobacillus Drugs 0.000 description 3
- -1 lipid peroxide Chemical class 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- 230000010355 oscillation Effects 0.000 description 3
- 230000017448 oviposition Effects 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 230000004083 survival effect Effects 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 208000013327 true hermaphroditism Diseases 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 2
- 241000588724 Escherichia coli Species 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 241000699666 Mus <mouse, genus> Species 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- GZCGUPFRVQAUEE-KCDKBNATSA-N aldehydo-D-galactose Chemical compound OC[C@@H](O)[C@H](O)[C@H](O)[C@@H](O)C=O GZCGUPFRVQAUEE-KCDKBNATSA-N 0.000 description 2
- 210000004958 brain cell Anatomy 0.000 description 2
- 244000309466 calf Species 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 235000014655 lactic acid Nutrition 0.000 description 2
- 239000004310 lactic acid Substances 0.000 description 2
- 230000031700 light absorption Effects 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 235000013336 milk Nutrition 0.000 description 2
- 239000008267 milk Substances 0.000 description 2
- 210000004080 milk Anatomy 0.000 description 2
- 230000001766 physiological effect Effects 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- AKYHKWQPZHDOBW-UHFFFAOYSA-N (5-ethenyl-1-azabicyclo[2.2.2]octan-7-yl)-(6-methoxyquinolin-4-yl)methanol Chemical compound OS(O)(=O)=O.C1C(C(C2)C=C)CCN2C1C(O)C1=CC=NC2=CC=C(OC)C=C21 AKYHKWQPZHDOBW-UHFFFAOYSA-N 0.000 description 1
- ZESRJSPZRDMNHY-YFWFAHHUSA-N 11-deoxycorticosterone Chemical compound O=C1CC[C@]2(C)[C@H]3CC[C@](C)([C@H](CC4)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1 ZESRJSPZRDMNHY-YFWFAHHUSA-N 0.000 description 1
- 108010023063 Bacto-peptone Proteins 0.000 description 1
- 241000244203 Caenorhabditis elegans Species 0.000 description 1
- XDTMQSROBMDMFD-UHFFFAOYSA-N Cyclohexane Chemical compound C1CCCCC1 XDTMQSROBMDMFD-UHFFFAOYSA-N 0.000 description 1
- 241000206602 Eukaryota Species 0.000 description 1
- 239000001576 FEMA 2977 Substances 0.000 description 1
- 241000287828 Gallus gallus Species 0.000 description 1
- JZNWSCPGTDBMEW-UHFFFAOYSA-N Glycerophosphorylethanolamin Natural products NCCOP(O)(=O)OCC(O)CO JZNWSCPGTDBMEW-UHFFFAOYSA-N 0.000 description 1
- WSMYVTOQOOLQHP-UHFFFAOYSA-N Malondialdehyde Chemical compound O=CCC=O WSMYVTOQOOLQHP-UHFFFAOYSA-N 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 208000012902 Nervous system disease Diseases 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 244000046052 Phaseolus vulgaris Species 0.000 description 1
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 102000005393 Sodium-Potassium-Exchanging ATPase Human genes 0.000 description 1
- 108010006431 Sodium-Potassium-Exchanging ATPase Proteins 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 102000019197 Superoxide Dismutase Human genes 0.000 description 1
- 108010012715 Superoxide dismutase Proteins 0.000 description 1
- 241000282898 Sus scrofa Species 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 230000001476 alcoholic effect Effects 0.000 description 1
- APUPEJJSWDHEBO-UHFFFAOYSA-P ammonium molybdate Chemical compound [NH4+].[NH4+].[O-][Mo]([O-])(=O)=O APUPEJJSWDHEBO-UHFFFAOYSA-P 0.000 description 1
- 239000011609 ammonium molybdate Substances 0.000 description 1
- 229940010552 ammonium molybdate Drugs 0.000 description 1
- 235000018660 ammonium molybdate Nutrition 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 238000005842 biochemical reaction Methods 0.000 description 1
- 230000036760 body temperature Effects 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 210000005252 bulbus oculi Anatomy 0.000 description 1
- FPPNZSSZRUTDAP-UWFZAAFLSA-N carbenicillin Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)C(C(O)=O)C1=CC=CC=C1 FPPNZSSZRUTDAP-UWFZAAFLSA-N 0.000 description 1
- 229960003669 carbenicillin Drugs 0.000 description 1
- 150000003943 catecholamines Chemical class 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- WORJEOGGNQDSOE-UHFFFAOYSA-N chloroform;methanol Chemical compound OC.ClC(Cl)Cl WORJEOGGNQDSOE-UHFFFAOYSA-N 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- LOUPRKONTZGTKE-UHFFFAOYSA-N cinchonine Natural products C1C(C(C2)C=C)CCN2C1C(O)C1=CC=NC2=CC=C(OC)C=C21 LOUPRKONTZGTKE-UHFFFAOYSA-N 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000003750 conditioning effect Effects 0.000 description 1
- 239000000356 contaminant Substances 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 229960003654 desoxycortone Drugs 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000002526 effect on cardiovascular system Effects 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 210000003608 fece Anatomy 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 235000021001 fermented dairy product Nutrition 0.000 description 1
- ODKNJVUHOIMIIZ-RRKCRQDMSA-N floxuridine Chemical compound C1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C(F)=C1 ODKNJVUHOIMIIZ-RRKCRQDMSA-N 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 235000020256 human milk Nutrition 0.000 description 1
- 210000004251 human milk Anatomy 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 235000021581 juice product Nutrition 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L magnesium chloride Substances [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 210000000214 mouth Anatomy 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 230000004768 organ dysfunction Effects 0.000 description 1
- LPMXVESGRSUGHW-HBYQJFLCSA-N ouabain Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1O[C@@H]1C[C@@]2(O)CC[C@H]3[C@@]4(O)CC[C@H](C=5COC(=O)C=5)[C@@]4(C)C[C@@H](O)[C@@H]3[C@@]2(CO)[C@H](O)C1 LPMXVESGRSUGHW-HBYQJFLCSA-N 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 150000008104 phosphatidylethanolamines Chemical class 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 235000020777 polyunsaturated fatty acids Nutrition 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 229960003110 quinine sulfate Drugs 0.000 description 1
- 150000003254 radicals Chemical class 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 239000012898 sample dilution Substances 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000007711 solidification Methods 0.000 description 1
- 230000008023 solidification Effects 0.000 description 1
- 235000013599 spices Nutrition 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000002562 thickening agent Substances 0.000 description 1
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 1
- 239000012137 tryptone Substances 0.000 description 1
- 210000001215 vagina Anatomy 0.000 description 1
- 238000003805 vibration mixing Methods 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
Landscapes
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention provides a kind of Lactobacillus salivarius (Lactobacillus salivarius), it is characterised in that the deposit number of described Lactobacillus salivarius is CGMCC No.6288.Present invention also offers a kind of functional food composition and preparation method thereof, including: the viable bacteria body of Lactobacillus salivarius as above is inactivated, obtains thalline material.Present invention also offers a kind of functional food composition, described functional food composition contains the viable bacteria body of Lactobacillus salivarius as above.Present invention also offers the Lactobacillus salivarius as above application in the functional food composition preparing slow down aging.Thalline material after the viable bacteria body of Lactobacillus salivarius provided by the invention and inactivation is respectively provided with effect of slow down aging.
Description
Technical field
The present invention relates to Lactobacillus salivarius (Lactobacillussalivarius) and application and functional food composition and preparation method thereof, specifically, relate to a kind of Lactobacillus salivarius, functional food composition containing this Lactobacillus salivarius and preparation method thereof, and the application that this Lactobacillus salivarius is in the functional food preparing slow down aging.
Background technology
Aging be body tissue, organ dysfunction with the age increase and occur degeneration change (Turnheim, 2003), be the general performance of the various biochemical reaction of body.Economic fast development, the progress of science and technology, make developed country and some developing countries step into aging society.The problem of an aging population has had become as global social problem.By the end of the year 2009, China 60 years old and above aging population reach 1.6714 hundred million, account for the 12.5% of total population, it is contemplated that be up to 2.43 hundred million to the year two thousand twenty.With advancing age and the aging of body, the disease relevant to aging, such as cardiovascular, nervous system disease, tumor, diabetes etc., threaten life-span and the quality of life of the mankind.Therefore, increasing research is absorbed in searching and is had the material of anti-aging effects and study its defying age mechanism.Current multiple natural product is it is verified that have anti-aging effects.But the material source with anti-aging effects being presently considered to be is comparatively single, separates purification cost high, and the medicine with anti-aging effects has again certain side effect, thus limiting the widespread adoption of these materials.Probiotic bacteria safety is higher, separates purification cost low, if it is possible to the anti-senescence function of clear and definite probiotic bacteria, and determine its mechanism of resisting senility, will very wide using probiotic bacteria as the prospect of anti-aging product.
Lactobacillus salivarius is the lactobacillus being widely present in human body and animal body.The oral cavity of human body, breast milk, woman vagina can detect Lactobacillus salivarius (the few equality of Lee, 1991;Olivaresetal., 2006).It is the ancestral home bacterium of human body, one of normal flora being present in human body；Except human body, be also separated to from the intestinals such as chicken, pig, rabbit Lactobacillus salivarius (Chen Guiyin, 2006;Tang Xiaoli, 2007;Luo Rui, 2011).And Lactobacillus salivarius is not belonging to common pathogenic bacterium, seldom having the report that Lactobacillus salivarius is diseases induced, therefore the lactobacillus salivarius strains that screening obtains all is carried out functional study by a lot of researchs, uses to the probiotic bacteria as human and animal.
Summary of the invention
It is an object of the invention to provide a kind of Lactobacillus salivarius with deferring senility.
To achieve these goals, on the one hand, the invention provides a kind of Lactobacillus salivarius (Lactobacillussalivarius), it is characterised in that the deposit number of described Lactobacillus salivarius is CGMCCNo.6288.
Second aspect, the invention provides a kind of method preparing functional food composition, and described method includes: is inactivated by the viable bacteria body of Lactobacillus salivarius as above, obtains thalline material.
The third aspect, the invention provides a kind of functional food composition prepared by method as above.
Fourth aspect, the invention provides a kind of functional food composition, it is characterised in that described functional food composition contains the viable bacteria body of Lactobacillus salivarius as above.
5th aspect, the invention provides the Lactobacillus salivarius as above application in the functional food composition preparing slow down aging.
Thalline material after the viable bacteria body of Lactobacillus salivarius provided by the invention and inactivation is respectively provided with effect of slow down aging.
Other features and advantages of the present invention will be described in detail in detailed description of the invention part subsequently.
Biological deposits
The Lactobacillus salivarius (Lactobacillussalivarius) of the present invention, it is deposited in (address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, China Committee for Culture Collection of Microorganisms's common micro-organisms center on June 25th, 2012, Institute of Microorganism, Academia Sinica, postcode: 100101) (depositary institution is abbreviated as CGMCC), deposit number is CGMCCNo.6288.
Detailed description of the invention
Hereinafter the specific embodiment of the present invention is described in detail.It should be appreciated that detailed description of the invention described herein is merely to illustrate and explains the present invention, it is not limited to the present invention.
On the one hand, the invention provides a kind of Lactobacillus salivarius (Lactobacillussalivarius), the deposit number of this Lactobacillus salivarius is CGMCCNo.6288.
The Lactobacillus salivarius of the present invention is isolatable from the feces of Bama County of Guangxi long lived elder.
Lactobacillus salivarius provided by the invention can produce the viable bacteria body of a large amount of Lactobacillus salivarius through cultivating, and the not special requirement of the method for described cultivation, as long as can make described Lactobacillus salivarius breed, for instance can by 107The inoculation of magnitude is in lactobacillus culture medium, and under anaerobism or aerobic condition, after cultivating 8-72 hour, obtains culture fluid at the temperature of 15-38 DEG C.Described lactobacillus culture medium can be the culture medium that known various applicable lactobacilluss are cultivated, can be such as milk and/or " lactic acid bacteria biological basis and application " (Yang Jiebin, light industry publishing house, 1996 publish) described in lactic acid bacteria (MRS) culture medium.
The present invention can separate the viable bacteria body of the Lactobacillus salivarius in above-mentioned culture fluid further, the method of described separation has no particular limits, as long as thalline can be enriched with from culture fluid, such as can pass through method that is centrifugal and/or that filter to realize, described centrifugal and described filtration condition can be known condition, and the present invention does not repeat them here.
Second aspect, the invention provides a kind of method preparing functional food composition, and described method includes: is inactivated by the viable bacteria body of Lactobacillus salivarius as above, obtains thalline material.
The present inventor has been surprisingly found that under study for action, the thalline material obtained after inactivateing 0.5-1.5h at 65-85 DEG C, has effect of more significantly slow down aging.Therefore, the condition of inactivation preferably includes: temperature is 65-85 DEG C, and the time is 0.5-1.5h.
The inventive method also includes adding in food thalline material to.
According to the present invention, although adding in food by thalline material, the purpose of the present invention can be realized, namely the effect of slow down aging is played, but under preferable case, with the gross weight of functional food composition for benchmark, the addition of thalline material is 10-70 weight %, more preferably 30-50 weight %.Under above-mentioned preferable case, the effect of the slow down aging of functional food composition is more notable.
In the present invention, food can be any type of food, for instance juice product, milk product, bean product etc..Food can also be different according to the difference of edible object.Described functional food composition can also contain conventional additive, for instance spice, mineral, vitamin, stabilizer, thickening agent, preservative etc..
The third aspect, the invention provides a kind of functional food composition prepared by method as above.
Fourth aspect, the invention provides a kind of functional food composition, and this functional food composition contains the viable bacteria body of Lactobacillus salivarius as above.
According to the present invention, although functional food composition contains the viable bacteria body of Lactobacillus salivarius as above, the purpose of the present invention can be realized, namely play the effect of slow down aging.But under preferable case, with the gross weight of functional food composition for benchmark, the content of the viable bacteria body of Lactobacillus salivarius is 105-1010CFU/g, more preferably 107-109CFU/g.Under above-mentioned preferable case, the effect of the slow down aging of functional food composition is more notable.
The functional food composition meeting above-mentioned requirements can include the viable bacteria body etc. of the Lactobacillus salivarius of the culture fluid of Lactobacillus salivarius (fermented dairy product such as prepared), separation through the fermentation of this Lactobacillus salivarius.
In the present invention, functional food composition is possibly together with food, and food is not as it was previously stated, repeat them here.
5th aspect, the invention provides the Lactobacillus salivarius as above application in the functional food composition preparing slow down aging.
The preferred embodiment of the present invention described in detail above; but, the present invention is not limited to the detail in above-mentioned embodiment, in the technology concept of the present invention; technical scheme can being carried out multiple simple variant, these simple variant belong to protection scope of the present invention.
It is further to note that, each concrete technical characteristic described in above-mentioned detailed description of the invention, in reconcilable situation, it is possible to be combined by any suitable mode, in order to avoid unnecessary repetition, various possible compound modes are no longer illustrated by the present invention separately.
Additionally, can also carry out combination in any between the various different embodiment of the present invention, as long as it is without prejudice to the thought of the present invention, it should be considered as content disclosed in this invention equally.
Embodiment
The present invention is further illustrated for below example, but and is not so limited the present invention.
(1) adopt nematicide as study subject
Nematicide is growth, apoptosis, old and feeble area research model organism the most clearly, its life cycle is short, and environment sensitive to external world, and owing to the homology of old and feeble signal is high, mammal and human senility are had directly with reference to (GreerandBrunet, 2008) by the research of unicellular lower eukaryote.Therefore, adopt nematicide as study subject in the following Examples and Comparative Examples.
In following embodiment and comparative example:
Nematicide is Caenorhabditis elegans worm strain, wild type CaenorhabditiselegansN2, purchased from American nematicide hereditism center (CaenorhabditisGeneticsCenter(CGC)).
Strain: nematicide standard chow escherichia coli (E.coli) OP50, purchased from American nematicide hereditism center；Lactobacillus salivarius A is that (this bacterial strain is deposited in (address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, China Committee for Culture Collection of Microorganisms's common micro-organisms center on June 25th, 2012 for the Lactobacillus salivarius of the present invention, Institute of Microorganism, Academia Sinica, postcode: 100101) (depositary institution is abbreviated as CGMCC), deposit number is CGMCCNo.6288).
Nematicide basic operation method carries out with reference to Wormbook standard eelworm culture method.
1M kaliumphosphate buffer (pH6.0): 108.3gKH2PO4, 46.8gK2HPO4·3H2O, distilled water is settled to 1L, and 121 DEG C of sterilizing 15min are standby.
NGM culture medium: 3.0gNaCl, 20g agar powder, 2.5g bacto peptone, add distilled water 970mL, 121 DEG C of sterilizing 15min, be placed in 60 DEG C of water-baths, under aseptic condition, add following component: 1mL1MMgSO4, 1mL1MCaCl2, 25mL1M kaliumphosphate buffer, the alcoholic solution of 3.2mL5mg/mL cholesterol；After mixing, pour plate.After flat board solidifies, being placed in room temperature 2 days, volatilization moisture and inspection flat board are placed in 4 DEG C of seal boxs without living contaminants and save backup.
MNGM culture medium: aseptically add before NGM culture medium pour plate through the FUdR of filtration sterilization, Carbenicillin to final concentration respectively 50 μMs, 0.5mM, mixing pour plate, solidification is placed on 2 days volatilization moisture of room temperature, rear plate can be coated with bacterium and use or be placed in 4 DEG C of seal boxs and preserve, used complete in 1 month.
M9 buffer: 3gKH2PO4, 15.12gNa2HPO4·12H2O, 5gNaCl, add distilled water and be settled to 1L, 121 DEG C of sterilizing 15min, aseptic addition 1mL1MMgSO after being cooled to room temperature4, mix standby.
Nematicide lysate: 1mL8MNaOH, 0.6mL9% liquor natrii hypochloritis, the aseptic ddH of 3.4mL2O, uses after mixing, matching while using.
LB culture medium: tryptone 10g, yeast leaching powder 5g, NaCl10g(solid medium adds 15g agar), add 950mL distilled water, regulate pH to 7.0,121 DEG C of sterilizing 15min standby.
MRS culture medium: be purchased from Beijing overpass company.
Embodiment 1.1
The present embodiment is for illustrating effect of the Lactobacillus salivarius slow down aging of the present invention.
(1) escherichia coli OP50 is seeded to LB fluid medium with the inoculum concentration of 1%, in 37 DEG C of 220rpm shaken cultivation 8h of constant-temperature table to late log phase, obtains escherichia coli OP50 culture fluid；Lactobacillus salivarius A is seeded to MRS fluid medium with the inoculum concentration of 1%, cultivates 12h to late log phase in 37 DEG C of nitrogen charging anaerobism bottle, obtain Lactobacillus salivarius A culture fluid.
Respectively by the culture fluid of above-mentioned escherichia coli OP50 and Lactobacillus salivarius A, collecting thalline with the centrifugal 15min of 4000 × g, wash twice by M9 buffer repeated centrifugation, 4 DEG C of centrifugal 15min of 16000 × g, remove whole supernatant, weigh thalline weight in wet base afterwards.Thalline is resuspended with a certain amount of M9, make the final concentration of 400mg/mL of thalline, be sub-packed in 1.5mL centrifuge tube, as concentration thalline.
Respectively above two is concentrated thalline 75 DEG C heat inactivation 1h, after cooling, escherichia coli OP50 and Lactobacillus salivarius A is mixed to get mixture by the weight ratio (namely Lactobacillus salivarius is 40 weight %) of 3:2, mixture is suspended in M9 buffer according to the ratio of 400mg/mL, coat the mNGM planar surface of diameter 60mm, making every flat board biomass is 10mg, totally 6 pieces of flat boards, 4 DEG C of sealings save backup.
(2) in diameter 90mmNGM media surface, even spread 200 μ L escherichia coli OP50 culture fluid, to cultivate 10h for 37 DEG C, go down to posterity as nematicide and use flat board, 4 DEG C of sealings save backup.
Aseptically, having, from growth, the fritter culture medium cutting about 1cm × 1cm the NGM flat board of nematicide with scalpel, facing down is buckled in nematicide and goes down to posterity with, on flat board, being placed in 25 DEG C of cultivation, and 60mm plate goes down to posterity once in general 7 days, keeps nematicide active.
Take adult after going down to posterity and be mostly in the flat board in egg-laying season for nematicide synchronization:
Adding 2mLM9 buffer in flat board, blow and beat with aseptic straw, sweep away the polypide on flat board as far as possible, polypide is transferred to 1.5mL centrifuge tube, 800 × g is centrifuged 1min, abandons supernatant and collects polypide, adds 1mLM9 recentrifuge washing polypide, exhausts supernatant as far as possible.
Adding the fresh nematicide lysate of 1mL, vortex oscillation 10s at once, 800 × g is centrifuged 1min, and pipettor siphons away supernatant.
Being added immediately 1mLM9 and wash polypide, 800 × g is centrifuged 1min, double, removes residual lysate.Finally in pipe, residue about 100 μ L contain the M9 suspension of worm's ovum.
Use aseptic straw to be dripped by worm's ovum in the NGM flat board of coating OP50, cultivate 48h, obtain the nematicide of synchronization 48h for 25 DEG C.
(3) by the nematicide of synchronization 48h, be placed in stereomicroscope observation, use platinum shovel picking tocostome be transparent semi-moon shaped L4 phase hermaphroditism worm on the mNGM flat board that step (1) obtains, every piece of flat board picking 10, flat board is placed in 25 DEG C of cultivations.Observe nematicide survival state every day, do not have movable reaction then to think that polypide is dead as platinum shovel is touched by polypide.Get rid of hatching missing, internal and unnatural death nematicide.Whole nematicide death are then tested and are terminated.Calculate nematicide average life in Table 1.
Embodiment 1.2
The present embodiment is for illustrating effect of the Lactobacillus salivarius slow down aging of the present invention.
(1) escherichia coli OP50 is seeded to LB fluid medium with the inoculum concentration of 1%, in 37 DEG C of 220rpm shaken cultivation 8h of constant-temperature table to late log phase, obtains escherichia coli OP50 culture fluid；Lactobacillus salivarius A is seeded to MRS fluid medium with the inoculum concentration of 1%, cultivates 12h to late log phase in 37 DEG C of nitrogen charging anaerobism bottle, obtain Lactobacillus salivarius A culture fluid.
Respectively by the culture fluid of above-mentioned escherichia coli OP50 and Lactobacillus salivarius A, collecting thalline with the centrifugal 15min of 4000 × g, wash twice by M9 buffer repeated centrifugation, 4 DEG C of centrifugal 15min of 16000 × g, remove whole supernatant, weigh thalline weight in wet base afterwards.Thalline is resuspended with a certain amount of M9, make the final concentration of 400mg/mL of thalline, be sub-packed in 1.5mL centrifuge tube, as concentration thalline.
Respectively above two is concentrated thalline 65 DEG C heat inactivation 1.5h, after cooling, escherichia coli OP50 and Lactobacillus salivarius A is mixed to get mixture by the weight ratio (namely Lactobacillus salivarius is 30 weight %) of 7:3, mixture is suspended in M9 buffer according to the ratio of 400mg/mL, coat the mNGM planar surface of diameter 60mm, making every flat board biomass is 10mg, totally 6 pieces of flat boards, 4 DEG C of sealings save backup.
(2) in diameter 90mmNGM media surface, even spread 200 μ L escherichia coli OP50 culture fluid, to cultivate 10h for 37 DEG C, go down to posterity as nematicide and use flat board, 4 DEG C of sealings save backup.
Aseptically, having, from growth, the fritter culture medium cutting about 1cm × 1cm the NGM flat board of nematicide with scalpel, facing down is buckled in nematicide and goes down to posterity with, on flat board, being placed in 25 DEG C of cultivation, and 60mm plate goes down to posterity once in general 7 days, keeps nematicide active.
Take adult after going down to posterity and be mostly in the flat board in egg-laying season for nematicide synchronization:
Adding 2mLM9 buffer in flat board, blow and beat with aseptic straw, sweep away the polypide on flat board as far as possible, polypide is transferred to 1.5mL centrifuge tube, 800 × g is centrifuged 1min, abandons supernatant and collects polypide, adds 1mLM9 recentrifuge washing polypide, exhausts supernatant as far as possible.
Adding the fresh nematicide lysate of 1mL, vortex oscillation 10s at once, 800 × g is centrifuged 1min, and pipettor siphons away supernatant.
Being added immediately 1mLM9 and wash polypide, 800 × g is centrifuged 1min, double, removes residual lysate.Finally in pipe, residue about 100 μ L contain the M9 suspension of worm's ovum.
Use aseptic straw to be dripped by worm's ovum in the NGM flat board of coating OP50, cultivate 48h, obtain the nematicide of synchronization 48h for 25 DEG C.
(3) by the nematicide of synchronization 48h, be placed in stereomicroscope observation, use platinum shovel picking tocostome be transparent semi-moon shaped L4 phase hermaphroditism worm on the mNGM flat board that step (1) obtains, every piece of flat board picking 10, flat board is placed in 25 DEG C of cultivations.Observe nematicide survival state every day, do not have movable reaction then to think that polypide is dead as platinum shovel is touched by polypide.Get rid of hatching missing, internal and unnatural death nematicide.Whole nematicide death are then tested and are terminated.Calculate nematicide average life in Table 1.
Embodiment 1.3
The present embodiment is for illustrating effect of the Lactobacillus salivarius slow down aging of the present invention.
(1) escherichia coli OP50 is seeded to LB fluid medium with the inoculum concentration of 1%, in 37 DEG C of 220rpm shaken cultivation 8h of constant-temperature table to late log phase, obtains escherichia coli OP50 culture fluid；Lactobacillus salivarius A is seeded to MRS fluid medium with the inoculum concentration of 1%, cultivates 12h to late log phase in 37 DEG C of nitrogen charging anaerobism bottle, obtain Lactobacillus salivarius A culture fluid.
Respectively by the culture fluid of above-mentioned escherichia coli OP50 and Lactobacillus salivarius A, collecting thalline with the centrifugal 15min of 4000 × g, wash twice by M9 buffer repeated centrifugation, 4 DEG C of centrifugal 15min of 16000 × g, remove whole supernatant, weigh thalline weight in wet base afterwards.Thalline is resuspended with a certain amount of M9, make the final concentration of 400mg/mL of thalline, be sub-packed in 1.5mL centrifuge tube, as concentration thalline.
Respectively above two is concentrated thalline 85 DEG C heat inactivation 0.5h, after cooling, escherichia coli OP50 and Lactobacillus salivarius A is mixed to get mixture by the weight ratio (namely Lactobacillus salivarius is 50 weight %) of 1:1, mixture is suspended in M9 buffer according to the ratio of 400mg/mL, coat the mNGM planar surface of diameter 60mm, making every flat board biomass is 10mg, totally 6 pieces of flat boards, 4 DEG C of sealings save backup.
(2) in diameter 90mmNGM media surface, even spread 200 μ L escherichia coli OP50 culture fluid, to cultivate 10h for 37 DEG C, go down to posterity as nematicide and use flat board, 4 DEG C of sealings save backup.
Aseptically, having, from growth, the fritter culture medium cutting about 1cm × 1cm the NGM flat board of nematicide with scalpel, facing down is buckled in nematicide and goes down to posterity with, on flat board, being placed in 25 DEG C of cultivation, and 60mm plate goes down to posterity once in general 7 days, keeps nematicide active.
Take adult after going down to posterity and be mostly in the flat board in egg-laying season for nematicide synchronization:
Adding 2mLM9 buffer in flat board, blow and beat with aseptic straw, sweep away the polypide on flat board as far as possible, polypide is transferred to 1.5mL centrifuge tube, 800 × g is centrifuged 1min, abandons supernatant and collects polypide, adds 1mLM9 recentrifuge washing polypide, exhausts supernatant as far as possible.
Adding the fresh nematicide lysate of 1mL, vortex oscillation 10s at once, 800 × g is centrifuged 1min, and pipettor siphons away supernatant.
Being added immediately 1mLM9 and wash polypide, 800 × g is centrifuged 1min, double, removes residual lysate.Finally in pipe, residue about 100 μ L contain the M9 suspension of worm's ovum.
Use aseptic straw to be dripped by worm's ovum in the NGM flat board of coating OP50, cultivate 48h, obtain the nematicide of synchronization 48h for 25 DEG C.
(3) by the nematicide of synchronization 48h, be placed in stereomicroscope observation, use platinum shovel picking tocostome be transparent semi-moon shaped L4 phase hermaphroditism worm on the mNGM flat board that step (1) obtains, every piece of flat board picking 10, flat board is placed in 25 DEG C of cultivations.Observe nematicide survival state every day, do not have movable reaction then to think that polypide is dead as platinum shovel is touched by polypide.Get rid of hatching missing, internal and unnatural death nematicide.Whole nematicide death are then tested and are terminated.Calculate nematicide average life in Table 1.
Embodiment 1.4
The present embodiment is for illustrating effect of the Lactobacillus salivarius slow down aging of the present invention.
Nematicide life test is carried out according to the method for embodiment 1.1, the difference is that, in step (1), after inactivation cooling, escherichia coli OP50 and Lactobacillus salivarius A is mixed by the weight ratio (namely Lactobacillus salivarius is 70 weight %) of 3:7.Calculate nematicide average life in Table 1.
Embodiment 1.5
The present embodiment is for illustrating effect of the Lactobacillus salivarius slow down aging of the present invention.
Nematicide life test is carried out according to the method for embodiment 1.1, the difference is that, in step (1), after inactivation cooling, escherichia coli OP50 and Lactobacillus salivarius A is mixed by the weight ratio (namely Lactobacillus salivarius is 10 weight %) of 9:1.Calculate nematicide average life in Table 1.
Embodiment 1.6
The present embodiment is for illustrating effect of the Lactobacillus salivarius slow down aging of the present invention.
Nematicide life test is carried out according to the method for embodiment 1.1, the difference is that, inactivation temperature is 90 DEG C, and inactivation time is 0.3h.Calculate nematicide average life in Table 1.
Embodiment 1.7
The present embodiment is for illustrating effect of the Lactobacillus salivarius slow down aging of the present invention.
Nematicide life test is carried out according to the method for embodiment 1.1, the difference is that, inactivation temperature is 60 DEG C, and inactivation time is 2.0h.Calculate nematicide average life in Table 1.
Comparative example 1.1
Nematicide life test is carried out according to the method for embodiment 1.1, the difference is that, substitute Lactobacillus salivarius A with bacterial strain disclosed in CN101538545A.Calculate nematicide average life in Table 1.
Comparative example 1.2
Nematicide life test is carried out according to the method for embodiment 1.1, the difference is that, after inactivation cooling, escherichia coli OP50 is suspended in M9 buffer according to the ratio of 400mg/mL, coats mNGM planar surface, that is, mNGM planar surface is only coated with the escherichia coli OP50 after inactivation cools down.Calculate nematicide average life in Table 1.
Table 1
| Nematicide average life (my god) | |
| Embodiment 1.1 | 24.1±0.4 |
| Embodiment 1.2 | 23.1±0.6 |
| Embodiment 1.3 | 24.3±0.5 |
| Embodiment 1.4 | 18.9±0.4 |
| Embodiment 1.5 | 18.7±0.9 |
| Embodiment 1.6 | 19.1±0.2 |
| Embodiment 1.7 | 20.0±0.3 |
| Comparative example 1.1 | 17.8±0.2 |
| Comparative example 1.2 | 17.2±1.0 |
Embodiment 1.1 is compared it can be seen that the Lactobacillus salivarius of the present invention has effect of slow down aging with comparative example 1.1 and comparative example 1.2 respectively, is used for after inactivation feeding nematicide, it is possible to the notable life-span extending nematicide.
Embodiment 1.1 is compared can be seen that with embodiment 1.4 and embodiment 1.5 respectively, after inactivation cooling, escherichia coli OP50 and Lactobacillus salivarius A is mixed, with the gross weight of mixture for benchmark, the addition of Lactobacillus salivarius A is 30-50 weight %, it is possible to the life-span of more significantly prolongation nematicide；Embodiment 1.1 is compared with embodiment 1.6 and embodiment 1.7 respectively it can be seen that inactivate 0.5-1.5h at 65-85 DEG C, be more beneficial for extending the life-span of nematicide.
(2) adopt mice as study subject
In following embodiment and comparative example:
Male Kunming strain mice: tie up company of tonneau China, body weight 20 ± 2g purchased from Beijing.
Lactobacillus salivarius A is that (this bacterial strain is deposited in (address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, China Committee for Culture Collection of Microorganisms's common micro-organisms center on June 25th, 2012 for the Lactobacillus salivarius of the present invention, Institute of Microorganism, Academia Sinica, postcode: 100101) (depositary institution is abbreviated as CGMCC), deposit number is CGMCCNo.6288).
MRS culture medium: be purchased from Beijing overpass company.
Coomassie brilliant blue G250: be purchased from Shanghai, Shanghai space biology company limited.
Iipofuscin content, brain MAO-B activity, brain SOD activity and brain Na+/K+The mensuration of-atpase activity is referring to " anti-aging effects of Water-soluable Polysaccharide from Rhizoma Dioscoreae Opposite on Mice " (Zhan Tong, Tao Jing, Wang Shuru, pharmacy is in progress, 23 volume the 6th phases in 1999):
Iipofuscin assay: take liver, 5% homogenate is made with chloroform-methanol mixed liquor, 40 DEG C of tepidarium 5 minutes, being centrifuged 10 minutes with 3000rpm, take supernatant and survey fluorescence, transmitting wavelength is 435nm, excitation wavelength is 365nm, put sensitivity × 1, slit 10nm, be 50 by the quinine sulfate conditioning instrumentation fluorescence intensity of 0.1 μ g/mL.
Brain MAO-B(B MAO-B B) determination of activity: take cerebral tissue, add the phosphate buffer (0.1mol/LK of ice-cold 0.2mol/LpH7.42HPO480.2ml with 0.1mol/LKH2PO419.8ml mixing), ultrasonic homogenate 2 times, it is centrifuged 10 minutes with 3000rpm in 4 DEG C, precipitation is suspended in PBS liquid (pH7.4), the vibration mixing of water-bath immediately, is centrifuged 10 minutes with 3000rpm, cyclohexane layer is surveyed in 242nm place light absorption value, crude enzyme liquid protein concentration Coomassie brilliant blue G250 measures, and makes standard protein with bSA, and enzyme activity unit is defined as the enzyme amount that 37 DEG C of optical density producing 0.01/3h change.
Brain SOD(superoxide dismutase) determination of activity: take cerebral tissue, tissue sample measures and adopts the SOD ultramicron of diease occurrence teaching and research group of Nanjing Railway College of Medicine quickly to measure reagent, enzymatic activity represents with U/mg enzyme amount, computing formula: [(control tube optical density-mensuration pipe optical density)/control tube optical density]/50/100 × sample dilution/sample protein concentration.Sample protein concentration Coomassie brilliant blue G250 measures, and calf serum makes standard protein.
Brain Na+/K+-atpase activity measures: takes cerebral tissue 0.1g, is rapidly added 1ml extracting solution and (includes sucrose 250mmol/L, histidine 30mmol/L, EDTA5mmol/L, desoxycortone 10 weight %.Ice bath homogenate, three layers filtered through gauze, obtain crude enzyme liquid, protein concentration Coomassie brilliant blue G250 measures.Enzymatic reaction is divided into two groups: A group, adds the medium A TP liquid (MgCl of 0.4mL26mmol/L, NaCl100mmol/L, KCl20mmol/L, Tris30mmol/L, ATP3mmol/L).50 μ L dielectric fluid (MgCl26mmol/L, NaCl100mmol/L, KCl20mmol/L, Tris30mmol/L) and crude enzyme liquid 50 μ L；B group, adds Quabain and suppresses Na+/K+-ATP enzyme, replaces 50 μ L dielectric fluids with the 5mmol/LQuabain of 50 μ L, and all the other are identical.37 DEG C of water-baths are reacted respectively 20 minutes, add 30% ice-cold trichloroacetic acid 0.3mL, move in ice bath, be centrifuged 5 minutes with 1500rpm, take supernatant 0.5ml, survey its content of inorganic phosphorus.Step is: takes 0.5mL and measures liquid, adds the 2mL developer (FeSO of 1 volume 10% ammonium molybdate+9 volume freshly prepared 9.15%4·7H2O) mixing with 2mL distilled water, 25 DEG C are reacted 8 minutes, survey light absorption value in 652nm place, make standard curve with standard phosphorus solution, try to achieve content of inorganic phosphorus, last Na+/K+The content of inorganic phosphorus that-ATP enzyme is decomposed is the amount of A group-B group.Enzymatic activity is expressed as a μm olpi/h rmg albumen, and protein concentration Coomassie brilliant blue G250 measures, and calf serum makes standard protein.
Embodiment 2.1
The present embodiment is for illustrating effect of the Lactobacillus salivarius slow down aging of the present invention.
(1) Lactobacillus salivarius A aseptic inoculation ring is inoculated in the MRS culture medium of 10L, and under anaerobic, after cultivating 12 hours at the temperature of 37 DEG C, obtains culture fluid.Supernatant discarded after being centrifuged 10 minutes under the speed of 4000 × g by the culture fluid obtained, obtains the viable bacteria body of Lactobacillus salivarius A.
(2) choosing 15 Male Kunming strain mice immediately, every day, retrobulbar injection D-galactose 0.12mg/g, and fed containing 1 × 109The drinking water of the viable bacteria body of the Lactobacillus salivarius A of CFU/g, continuous one month.Survey the Iipofuscin content of Male Kunming strain mice, brain MAO-B activity, brain SOD activity and brain Na+/K+-atpase activity, result is in Table 2.
Embodiment 2.2
The present embodiment is for illustrating effect of the Lactobacillus salivarius slow down aging of the present invention.
(1) Lactobacillus salivarius A aseptic inoculation ring is inoculated in the MRS culture medium of 10L, and under anaerobic, after cultivating 12 hours at the temperature of 37 DEG C, obtains culture fluid.Supernatant discarded after being centrifuged 10 minutes under the speed of 4000 × g by the culture fluid obtained, obtains the viable bacteria body of Lactobacillus salivarius A.
(2) choosing 15 Male Kunming strain mice immediately, every day, retrobulbar injection D-galactose 0.12mg/g, and fed containing 1 × 108The drinking water of the viable bacteria body of the Lactobacillus salivarius A of CFU/g, continuous one month.Survey the Iipofuscin content of Male Kunming strain mice, brain MAO-B activity, brain SOD activity and brain Na+/K+-atpase activity, result is in Table 2.
Embodiment 2.3
The present embodiment is for illustrating effect of the Lactobacillus salivarius slow down aging of the present invention.
(1) Lactobacillus salivarius A aseptic inoculation ring is inoculated in the MRS culture medium of 10L, and under anaerobic, after cultivating 12 hours at the temperature of 37 DEG C, obtains culture fluid.Supernatant discarded after being centrifuged 10 minutes under the speed of 4000 × g by the culture fluid obtained, obtains the viable bacteria body of Lactobacillus salivarius A.
(2) choosing 15 Male Kunming strain mice immediately, every day, retrobulbar injection D-galactose 0.12mg/g, and fed containing 1 × 107The drinking water of the viable bacteria body of the Lactobacillus salivarius A of CFU/g, continuous one month.Survey the Iipofuscin content of Male Kunming strain mice, brain MAO-B activity, brain SOD activity and brain Na+/K+-atpase activity, result is in Table 2.
Embodiment 2.4
The present embodiment is for illustrating effect of the Lactobacillus salivarius slow down aging of the present invention.
Test according to the method for embodiment 2.1, the difference is that, in drinking water, the content of the viable bacteria body of Lactobacillus salivarius A is 1 × 1010CFU/g.Survey the Iipofuscin content of Male Kunming strain mice, brain MAO-B activity, brain SOD activity and brain Na+/K+-atpase activity, result is in Table 2.
Embodiment 2.5
The present embodiment is for illustrating effect of the Lactobacillus salivarius slow down aging of the present invention.
Test according to the method for embodiment 2.1, the difference is that, in drinking water, the content of the viable bacteria body of Lactobacillus salivarius A is 1 × 105CFU/g.Survey the Iipofuscin content of Male Kunming strain mice, brain MAO-B activity, brain SOD activity and brain Na+/K+-atpase activity, result is in Table 2.
Embodiment 2.6
The present embodiment is for illustrating effect of the Lactobacillus salivarius slow down aging of the present invention.
Test according to the method for embodiment 2.1, the difference is that, step (1) is obtained the viable bacteria body of Lactobacillus salivarius A heat inactivation 1h at 75 DEG C, obtains thalline material, being joined by this thalline material and be added without viable bacteria body in drinking water, in drinking water, the content of thalline material is 40 weight %.Survey the Iipofuscin content of Male Kunming strain mice, brain MAO-B activity, brain SOD activity and brain Na+/K+-atpase activity, result is in Table 2.
Embodiment 2.7
The present embodiment is for illustrating effect of the Lactobacillus salivarius slow down aging of the present invention.
Test according to the method for embodiment 2.1, the difference is that, step (1) is obtained the viable bacteria body of Lactobacillus salivarius A heat inactivation 1.5h at 65 DEG C, obtains thalline material, being joined by this thalline material and be added without viable bacteria body in drinking water, in drinking water, the content of thalline material is 50 weight %.Survey the Iipofuscin content of Male Kunming strain mice, brain MAO-B activity, brain SOD activity and brain Na+/K+-atpase activity, result is in Table 2.
Embodiment 2.8
The present embodiment is for illustrating effect of the Lactobacillus salivarius slow down aging of the present invention.
Test according to the method for embodiment 2.1, the difference is that, step (1) is obtained the viable bacteria body of Lactobacillus salivarius A heat inactivation 0.5h at 85 DEG C, obtains thalline material, being joined by this thalline material and be added without viable bacteria body in drinking water, in drinking water, the content of thalline material is 30 weight %.Survey the Iipofuscin content of Male Kunming strain mice, brain MAO-B activity, brain SOD activity and brain Na+/K+-atpase activity, result is in Table 2.
Embodiment 2.9
The present embodiment is for illustrating effect of the Lactobacillus salivarius slow down aging of the present invention.
Test according to the method for embodiment 2.6, the difference is that, in drinking water, the content of thalline material is 65 weight %.Survey the Iipofuscin content of Male Kunming strain mice, brain MAO-B activity, brain SOD activity and brain Na+/K+-atpase activity, result is in Table 2.
Embodiment 2.10
The present embodiment is for illustrating effect of the Lactobacillus salivarius slow down aging of the present invention.
Test according to the method for embodiment 2.6, the difference is that, in drinking water, the content of thalline material is 15 weight %.Survey the Iipofuscin content of Male Kunming strain mice, brain MAO-B activity, brain SOD activity and brain Na+/K+-atpase activity, result is in Table 2.
Embodiment 2.11
The present embodiment is for illustrating effect of the Lactobacillus salivarius slow down aging of the present invention.
Test according to the method for embodiment 2.6, the difference is that, inactivation temperature is 90 DEG C, and inactivation time is 0.3h.Survey the Iipofuscin content of Male Kunming strain mice, brain MAO-B activity, brain SOD activity and brain Na+/K+-atpase activity, result is in Table 2.
Embodiment 2.12
The present embodiment is for illustrating effect of the Lactobacillus salivarius slow down aging of the present invention.
Test according to the method for embodiment 2.6, the difference is that, inactivation temperature is 60 DEG C, and inactivation time is 2.0h.Survey the Iipofuscin content of Male Kunming strain mice, brain MAO-B activity, brain SOD activity and brain Na+/K+-atpase activity, result is in Table 2.
Comparative example 2.1
Test according to the method for embodiment 2.1, the difference is that, substitute Lactobacillus salivarius A with bacterial strain disclosed in CN101538545A.Survey the Iipofuscin content of Male Kunming strain mice, brain MAO-B activity, brain SOD activity and brain Na+/K+-atpase activity, result is in Table 2.
Comparative example 2.2
Test according to the method for embodiment 2.6, the difference is that, substitute Lactobacillus salivarius A with bacterial strain disclosed in CN101538545A.Survey the Iipofuscin content of Male Kunming strain mice, brain MAO-B activity, brain SOD activity and brain Na+/K+-atpase activity, result is in Table 2.
Comparative example 2.3
Test according to the method for embodiment 2.1, the difference is that, drinking water is added without strain.Survey the Iipofuscin content of Male Kunming strain mice, brain MAO-B activity, brain SOD activity and brain Na+/K+-atpase activity, result is in Table 2.
Comparative example 2.4
Test according to the method for embodiment 2.1, the difference is that, not injection of d-galactose, drinking water is added without strain.Survey the Iipofuscin content of Male Kunming strain mice, brain MAO-B activity, brain SOD activity and brain Na+/K+-atpase activity, result is in Table 2.
Table 2
The free radical produced in metabolic process can act on the polyunsaturated fatty acid in biological film phospholipid, produce lipid peroxide, such as malonaldehyde (MDA), and then macromolecular complex-lipofuscin (age pigment) is become with PHOSPHATIDYL ETHANOLAMINE and protein cross, this is a kind of mark of organism aging process.
In human brain, MAO-B activity increased with the age after 45 years old and increases, Fowier finds that the MAO-B activity in human brain 19 region was proportionate with the age, in cerebral tissue, MAO-B can cause changes of Catecholamine Content in brain disorderly with the change at age, promotes physiological activity to lack of proper care, thus causing old and feeble generation.
SOD is the internal oxygen radical removing enzyme that a class is important, but increases with the age, and the SOD activity in body tissue lowers gradually.
Na+/K+-ATP enzyme, also known as sodiumpotassium pump, is a kind of particularly important film enzyme being widely present in organism, and this enzyme plays an important role in ionic equilibrium inside and outside maintenance physiological activity, body temperature and homergy and cell membrane.During biological decay, it may appear that multiple biochemical change, wherein the change of function of brain cell is more apparent, and this is likely to relevant with brain cell energy reduction.Have been reported that, its Na in some tissues of old rats+/K+The younger Mus of-atpase activity decreases.
Comparative example 2.3 and comparative example 2.4 are compared it can be seen that after eyeball of mouse injection of d-galactose, the aging of mice can be accelerated.Embodiment 2.1 to embodiment 2.12 and comparative example 2.3 are compared it can be seen that the Lactobacillus salivarius of the present invention has effect of slow down aging；Embodiment 2.1 and comparative example 2.1 are compared, embodiment 2.6 is compared with comparative example 2.2 it can be seen that the Lactobacillus salivarius of the present invention has effect of more significantly slow down aging than bacterial strain disclosed in CN101538545A.
Embodiment 2.1 is compared with embodiment 2.4 and embodiment 2.5 respectively it can be seen that the content of the viable bacteria body of Lactobacillus salivarius is 10 in drinking water7-109CFU/g, is more beneficial for slow down aging；Embodiment 2.6 is compared with embodiment 2.9 and embodiment 2.10 respectively it can be seen that the content of the thalline material obtained after the viable bacteria body inactivation of Lactobacillus salivarius in drinking water is 30-50 weight %, be more beneficial for slow down aging；Embodiment 2.6 is compared with embodiment 2.11 and embodiment 2.12 respectively it can be seen that inactivate 0.5-1.5h at 65-85 DEG C, be more beneficial for slow down aging.
Thalline material after the viable bacteria body of Lactobacillus salivarius provided by the invention and inactivation is respectively provided with effect of slow down aging.
Claims (12)
1. a Lactobacillus salivarius (Lactobacillussalivarius), it is characterised in that the deposit number of described Lactobacillus salivarius is CGMCCNo.6288.
2. the method preparing functional food composition, described method includes: is inactivated by the viable bacteria body of the Lactobacillus salivarius described in claim 1, obtains thalline material.
3. method according to claim 2, wherein, the condition of described inactivation includes: temperature is 65-85 DEG C, and the time is 0.5-1.5h.
4. according to the method in claim 2 or 3, wherein, described method also includes, and is added in food by described thalline material.
5. method according to claim 4, wherein, with the gross weight of described functional food composition for benchmark, the addition of described thalline material is 10-70 weight %.
6. method according to claim 4, wherein, with the gross weight of described functional food composition for benchmark, the addition of described thalline material is 30-50 weight %.
7. the functional food composition that prepared by the method described in any one in claim 2-6.
8. a functional food composition, it is characterised in that described functional food composition contains the viable bacteria body of the Lactobacillus salivarius described in claim 1.
9. functional food composition according to claim 8, wherein, with the gross weight of described functional food composition for benchmark, the content of the viable bacteria body of described Lactobacillus salivarius is 105-1010CFU/g。
10. functional food composition according to claim 8, wherein, with the gross weight of described functional food composition for benchmark, the content of the viable bacteria body of described Lactobacillus salivarius is 107-109CFU/g。
11. functional food composition described in any one in-10 according to Claim 8, wherein, described functional food composition is possibly together with food.
12. the application that the Lactobacillus salivarius described in claim 1 is in the functional food composition preparing slow down aging.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210572842.5A CN103897998B (en) | 2012-12-25 | 2012-12-25 | Lactobacillus salivarius and application and functional food composition and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210572842.5A CN103897998B (en) | 2012-12-25 | 2012-12-25 | Lactobacillus salivarius and application and functional food composition and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103897998A CN103897998A (en) | 2014-07-02 |
| CN103897998B true CN103897998B (en) | 2016-06-29 |
Family
ID=50989577
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210572842.5A Active CN103897998B (en) | 2012-12-25 | 2012-12-25 | Lactobacillus salivarius and application and functional food composition and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103897998B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105395575A (en) * | 2015-11-19 | 2016-03-16 | 徐州一统食品工业有限公司 | Chewable tablet containing inactivated lactobacillus salivarius and preparation method thereof |
| CN110923172B (en) * | 2019-12-25 | 2023-05-09 | 江西仁仁健康微生态科技有限公司 | Lactobacillus salivarius with bowel relaxing effect and cholesterol reducing effect and application thereof |
| CN111297914A (en) * | 2020-02-24 | 2020-06-19 | 北京农学院 | Application of lactobacillus fermentation clear liquid in antioxidation or preparation of product for antioxidation and antioxidation product |
| CN114558037B (en) * | 2022-02-24 | 2023-08-15 | 同济大学 | Application of AKK and LS in preparation of anti-aging products for improving cognition level |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101221239B1 (en) * | 2005-07-07 | 2013-01-11 | (주)바이오벤 | Lactic acid bacteria culture of Mung bean and the preparation method of the same, and the cosmetic composition comprising the same |
| CN101538545B (en) * | 2009-04-08 | 2011-07-20 | 任发政 | Lactobacillus salivarius and food composition thereof |
| EP2392340B1 (en) * | 2010-06-01 | 2014-01-08 | GenMont Biotech Inc. | Novel lactobacillus strain, composition and use thereof for treating diabetes |
| ES2386914B1 (en) * | 2011-02-09 | 2013-06-19 | José Miguel De Las Heras Del Río | USE OF A LACTOBACILLUS BULGARICUS CEPA FOR THE TREATMENT OF IMMUNOSENESCENCE. |
-
2012
- 2012-12-25 CN CN201210572842.5A patent/CN103897998B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN103897998A (en) | 2014-07-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105296381B (en) | One bacillus subtilis CYY-25 and its application | |
| CN103897998B (en) | Lactobacillus salivarius and application and functional food composition and preparation method thereof | |
| CN105602997B (en) | Enterobacter cloacae prepare method and the application of biological nano elemental selenium | |
| CN102229889A (en) | Chlorella, its culture method and applications | |
| CN110438028A (en) | A kind of people pig source Bei Laisi bacillus GX-1 of degraded cellulose | |
| CN110184209A (en) | One plant of Lactobacillus rhamnosus that can reduce blood uric acid | |
| CN103952362A (en) | Citrus endophytic actinomycetes with antibacterial activity on various plant pathogens | |
| CN104531546A (en) | Pseudomonas putida SRPG-396 and salt-dissolving and growth-promoting application thereof | |
| CN106359454A (en) | Preparation method of empedobacter brevis pesticide | |
| CN102816719B (en) | Growth-promoting rhizobacteria SXH-2 and application thereof | |
| CN104046573B (en) | Bifidobacteria longum and application thereof, and functional food composition and preparation method thereof | |
| CN104928206B (en) | One plant of lactobacillus acidophilus with high cholate hydrolytic enzyme activities and its application | |
| CN105925493B (en) | One plant of bamboo parasitic fungus and its application in fermenting and producing hypocrellin | |
| Dong et al. | Enhanced biomass production and wastewater treatment in attached co-culture of Chlorella pyrenoidosa with nitrogen-fixing bacteria Azotobacter beijerinckii | |
| CN102827792A (en) | Plant growth-promoting rhizobacterium SXH-3 and application thereof | |
| CN102492631B (en) | Amora-producing Geotrichum candidum strain of high-yield exopolysaccharides, exopolysaccharides and volatile flavor compounds | |
| CN103451116B (en) | Pseudomonas fluorescens Y13, and preparation method and applications thereof | |
| CN101805714B (en) | Hexachloro cyclohexane degrading bacteria and application thereof in degraded hexachloro cyclohexane | |
| CN104031856B (en) | Lactobacillus casei and application and functional food composition and preparation method thereof | |
| CN104130949B (en) | A kind of the liquid deep layer fermenting culture medium and cultural method of rich iron Cordyceps militaris | |
| CN109874329A (en) | A kind of production butyric acid is dwelt bacillus faecalis and its cultural method and application | |
| CN103540535B (en) | Intracellular lipase producing strains and application thereof and screening method and using method | |
| CN102031277B (en) | Aurovertin metabolins and application thereof | |
| CN105018370A (en) | Soybean endophytes and application thereof | |
| CN105713855A (en) | Strains, application of strains, vaccine and preparation method of vaccine |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C41 | Transfer of patent application or patent right or utility model | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20161031 Address after: 100083 Haidian District Qinghua East Road, No. 17, Beijing Patentee after: China Agricultural University Address before: 100085, room 5, building 39, No. 203 West Road, Beijing, Haidian District Patentee before: Beijing Zhongtian Shenzhou Space Foods Technology Research Institute |
|
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20170418 Address after: 610000 Tianfu Square, Tianfu Square, hi tech Zone, Sichuan, Chengdu, China T3-35 Patentee after: TIBET TREASURE OF PLATEAU YAK DAIRY INDUSTRY CO., LTD. Address before: 100083 Haidian District Qinghua East Road, No. 17, Beijing Patentee before: China Agricultural University |
|
| CP02 | Change in the address of a patent holder | ||
| CP02 | Change in the address of a patent holder |
Address after: 851400 A in Tibet District of Lhasa economic and Technological Development Zone Qionggang Lin Road East Road No. 6 Patentee after: TIBET TREASURE OF PLATEAU YAK DAIRY INDUSTRY CO., LTD. Address before: 610000 Tianfu Square, Tianfu Square, hi tech Zone, Sichuan, Chengdu, China T3-35 Patentee before: TIBET TREASURE OF PLATEAU YAK DAIRY INDUSTRY CO., LTD. |
|
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20211014 Address after: 851400 No. 6, East 1st Road, linqionggang Road, zone a, Lhasa Economic and Technological Development Zone, Lhasa City, Tibet Autonomous Region Patentee after: TIBET TREASURE OF PLATEAU YAK DAIRY INDUSTRY Co.,Ltd. Patentee after: QINGHAI TREASURE OF PLATEAU YAK MILK Co.,Ltd. Patentee after: RUOERBAO TREASURE OF PLATEAU YAK MILK Co.,Ltd. Address before: 851400 No.6, East 1st Road, linqionggang Road, zone a, Lhasa Economic and Technological Development Zone, Tibet Patentee before: TIBET TREASURE OF PLATEAU YAK DAIRY INDUSTRY Co.,Ltd. |
|
| CP01 | Change in the name or title of a patent holder | ||
| CP01 | Change in the name or title of a patent holder |
Address after: 851400 No. 6, East 1st Road, linqionggang Road, zone a, Lhasa Economic and Technological Development Zone, Lhasa City, Tibet Autonomous Region Patentee after: TIBET TREASURE OF PLATEAU YAK DAIRY INDUSTRY Co.,Ltd. Patentee after: QINGHAI TREASURE OF PLATEAU YAK MILK CO.,LTD. Patentee after: Ruoergai Plateau Zhibao yak milk nutritional food Co.,Ltd. Address before: 851400 No. 6, East 1st Road, linqionggang Road, zone a, Lhasa Economic and Technological Development Zone, Lhasa City, Tibet Autonomous Region Patentee before: TIBET TREASURE OF PLATEAU YAK DAIRY INDUSTRY Co.,Ltd. Patentee before: QINGHAI TREASURE OF PLATEAU YAK MILK CO.,LTD. Patentee before: RUOERBAO TREASURE OF PLATEAU YAK MILK CO.,LTD. |