CN117448178B - Monascus purpureus CEWL and application thereof in preparation of liver protection products - Google Patents
Monascus purpureus CEWL and application thereof in preparation of liver protection products Download PDFInfo
- Publication number
- CN117448178B CN117448178B CN202311431255.9A CN202311431255A CN117448178B CN 117448178 B CN117448178 B CN 117448178B CN 202311431255 A CN202311431255 A CN 202311431255A CN 117448178 B CN117448178 B CN 117448178B
- Authority
- CN
- China
- Prior art keywords
- liver
- cewl
- strain
- monascus purpureus
- purpureus
- 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
- 244000113306 Monascus purpureus Species 0.000 title claims abstract description 37
- 235000002322 Monascus purpureus Nutrition 0.000 title claims abstract description 37
- 229940057059 monascus purpureus Drugs 0.000 title claims abstract description 37
- 210000004185 liver Anatomy 0.000 title claims abstract description 30
- 238000002360 preparation method Methods 0.000 title claims description 9
- 230000000694 effects Effects 0.000 claims abstract description 40
- 239000003814 drug Substances 0.000 claims abstract description 17
- 238000004321 preservation Methods 0.000 claims abstract description 5
- 206010067125 Liver injury Diseases 0.000 claims description 30
- 231100000753 hepatic injury Toxicity 0.000 claims description 28
- 108010081577 aldehyde dehydrogenase (NAD(P)+) Proteins 0.000 claims description 15
- 230000001476 alcoholic effect Effects 0.000 claims description 6
- 239000000126 substance Substances 0.000 claims description 4
- 241000228212 Aspergillus Species 0.000 claims description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 abstract description 87
- 241000306554 Aspergillus purpureus Species 0.000 abstract description 10
- JMZOMFYRADAWOG-UHFFFAOYSA-N methyl 7-methoxy-4-(7-methoxy-5-methoxycarbonyl-1,3-benzodioxol-4-yl)-1,3-benzodioxole-5-carboxylate Chemical compound COC(=O)C1=CC(OC)=C2OCOC2=C1C1=C2OCOC2=C(OC)C=C1C(=O)OC JMZOMFYRADAWOG-UHFFFAOYSA-N 0.000 abstract description 7
- 230000000593 degrading effect Effects 0.000 abstract description 3
- 241000699670 Mus sp. Species 0.000 description 29
- 241000228347 Monascus <ascomycete fungus> Species 0.000 description 13
- VZGDMQKNWNREIO-UHFFFAOYSA-N tetrachloromethane Chemical compound ClC(Cl)(Cl)Cl VZGDMQKNWNREIO-UHFFFAOYSA-N 0.000 description 12
- 102100036475 Alanine aminotransferase 1 Human genes 0.000 description 10
- 108010082126 Alanine transaminase Proteins 0.000 description 10
- 239000000047 product Substances 0.000 description 10
- 229940079593 drug Drugs 0.000 description 9
- 239000001963 growth medium Substances 0.000 description 8
- 210000005228 liver tissue Anatomy 0.000 description 8
- 102000007698 Alcohol dehydrogenase Human genes 0.000 description 7
- 108010021809 Alcohol dehydrogenase Proteins 0.000 description 7
- 244000061456 Solanum tuberosum Species 0.000 description 7
- 235000002595 Solanum tuberosum Nutrition 0.000 description 7
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 7
- 239000000049 pigment Substances 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 6
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 description 6
- 238000001514 detection method Methods 0.000 description 6
- 210000002784 stomach Anatomy 0.000 description 6
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 5
- 230000002496 gastric effect Effects 0.000 description 5
- 210000005229 liver cell Anatomy 0.000 description 5
- 238000010186 staining Methods 0.000 description 5
- 239000000725 suspension Substances 0.000 description 5
- 108010003415 Aspartate Aminotransferases Proteins 0.000 description 4
- 102000004625 Aspartate Aminotransferases Human genes 0.000 description 4
- 241000233866 Fungi Species 0.000 description 4
- 241000699666 Mus <mouse, genus> Species 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 4
- 238000012258 culturing Methods 0.000 description 4
- 239000003651 drinking water Substances 0.000 description 4
- 235000020188 drinking water Nutrition 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 230000006872 improvement Effects 0.000 description 4
- 230000004783 oxidative metabolism Effects 0.000 description 4
- 210000002966 serum Anatomy 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- 108090000340 Transaminases Proteins 0.000 description 3
- 230000002075 anti-alcohol Effects 0.000 description 3
- 238000003556 assay Methods 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 230000015556 catabolic process Effects 0.000 description 3
- 230000006378 damage Effects 0.000 description 3
- 238000006731 degradation reaction Methods 0.000 description 3
- 239000004006 olive oil Substances 0.000 description 3
- 235000008390 olive oil Nutrition 0.000 description 3
- 239000012188 paraffin wax Substances 0.000 description 3
- 238000002791 soaking Methods 0.000 description 3
- 239000012192 staining solution Substances 0.000 description 3
- 102000014898 transaminase activity proteins Human genes 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- HSSYVKMJJLDTKZ-UHFFFAOYSA-N 3-phenylphthalic acid Chemical group OC(=O)C1=CC=CC(C=2C=CC=CC=2)=C1C(O)=O HSSYVKMJJLDTKZ-UHFFFAOYSA-N 0.000 description 2
- 208000022309 Alcoholic Liver disease Diseases 0.000 description 2
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 2
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical compound C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- IKHGUXGNUITLKF-XPULMUKRSA-N acetaldehyde Chemical compound [14CH]([14CH3])=O IKHGUXGNUITLKF-XPULMUKRSA-N 0.000 description 2
- 238000003149 assay kit Methods 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 210000004556 brain Anatomy 0.000 description 2
- 210000005252 bulbus oculi Anatomy 0.000 description 2
- 238000005119 centrifugation Methods 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
- 239000003085 diluting agent Substances 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 238000010579 first pass effect Methods 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 210000001156 gastric mucosa Anatomy 0.000 description 2
- 231100000234 hepatic damage Toxicity 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 239000007928 intraperitoneal injection Substances 0.000 description 2
- 238000009630 liquid culture Methods 0.000 description 2
- 230000008818 liver damage Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
- 239000006041 probiotic Substances 0.000 description 2
- 230000000529 probiotic effect Effects 0.000 description 2
- 235000018291 probiotics Nutrition 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 208000037816 tissue injury Diseases 0.000 description 2
- 231100000331 toxic Toxicity 0.000 description 2
- 230000002588 toxic effect Effects 0.000 description 2
- 239000008096 xylene Substances 0.000 description 2
- 208000007848 Alcoholism Diseases 0.000 description 1
- 108020002663 Aldehyde Dehydrogenase Proteins 0.000 description 1
- 102000005369 Aldehyde Dehydrogenase Human genes 0.000 description 1
- 238000009631 Broth culture Methods 0.000 description 1
- 208000008964 Chemical and Drug Induced Liver Injury Diseases 0.000 description 1
- 206010072268 Drug-induced liver injury Diseases 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 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 1
- 208000027761 Hepatic autoimmune disease Diseases 0.000 description 1
- 206010019799 Hepatitis viral Diseases 0.000 description 1
- PCZOHLXUXFIOCF-UHFFFAOYSA-N Monacolin X Natural products C12C(OC(=O)C(C)CC)CC(C)C=C2C=CC(C)C1CCC1CC(O)CC(=O)O1 PCZOHLXUXFIOCF-UHFFFAOYSA-N 0.000 description 1
- 210000000683 abdominal cavity Anatomy 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000003044 adaptive effect Effects 0.000 description 1
- 201000007930 alcohol dependence Diseases 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 210000003855 cell nucleus Anatomy 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000001784 detoxification Methods 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- PTKRHFQQMJPPJN-UHFFFAOYSA-N dipotassium;oxido-(oxido(dioxo)chromio)oxy-dioxochromium;sulfuric acid Chemical compound [K+].[K+].OS(O)(=O)=O.[O-][Cr](=O)(=O)O[Cr]([O-])(=O)=O PTKRHFQQMJPPJN-UHFFFAOYSA-N 0.000 description 1
- 230000035622 drinking Effects 0.000 description 1
- YQGOJNYOYNNSMM-UHFFFAOYSA-N eosin Chemical compound [Na+].OC(=O)C1=CC=CC=C1C1=C2C=C(Br)C(=O)C(Br)=C2OC2=C(Br)C(O)=C(Br)C=C21 YQGOJNYOYNNSMM-UHFFFAOYSA-N 0.000 description 1
- 208000010706 fatty liver disease Diseases 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 239000000834 fixative Substances 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 210000002064 heart cell Anatomy 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- PCZOHLXUXFIOCF-BXMDZJJMSA-N lovastatin Chemical compound C([C@H]1[C@@H](C)C=CC2=C[C@H](C)C[C@@H]([C@H]12)OC(=O)[C@@H](C)CC)C[C@@H]1C[C@@H](O)CC(=O)O1 PCZOHLXUXFIOCF-BXMDZJJMSA-N 0.000 description 1
- 229960004844 lovastatin Drugs 0.000 description 1
- QLJODMDSTUBWDW-UHFFFAOYSA-N lovastatin hydroxy acid Natural products C1=CC(C)C(CCC(O)CC(O)CC(O)=O)C2C(OC(=O)C(C)CC)CC(C)C=C21 QLJODMDSTUBWDW-UHFFFAOYSA-N 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000004570 mortar (masonry) Substances 0.000 description 1
- 238000002703 mutagenesis Methods 0.000 description 1
- 231100000350 mutagenesis Toxicity 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 239000008213 purified water Substances 0.000 description 1
- 239000001054 red pigment Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000007790 scraping Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 229930000044 secondary metabolite Natural products 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 239000008399 tap water Substances 0.000 description 1
- 235000020679 tap water Nutrition 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000000451 tissue damage Effects 0.000 description 1
- 231100000827 tissue damage Toxicity 0.000 description 1
- 238000009423 ventilation Methods 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 201000001862 viral hepatitis Diseases 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/145—Fungal isolates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L31/00—Edible extracts or preparations of fungi; Preparation or treatment thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention discloses aspergillus purpureus CEWL and application thereof in preparing liver-protecting products, wherein the strain is named Monascus purpureus CESWGF001, is preserved in China center for type culture collection, has the preservation number of CCTCC NO: M20231512, has obvious effects of degrading alcohol and protecting liver, has liver-protecting effects comparable to or even better than the effects of the existing liver-protecting medicament bifendate, and can be used for preparing liver-protecting products.
Description
Technical Field
The invention belongs to the field of microorganisms, and in particular relates to aspergillus purpureus CEWL and application thereof in preparing liver protection products.
Background
Monascus is an important pigment-producing microorganism and is widely used worldwide. The strain comprises a plurality of strains, wherein 8 kinds of important monascus which are formally collected and encoded by the microbiological institute of China academy of sciences and the food institute of light industry department comprise monascus purpureus. Monascus purpureus (Monascus purpureus) is a saprophytic fungus, and is widely focused because of the capability of producing a large amount of natural red pigment, and is generally present in environments such as trees, soil, sediments and the like, and is now classified as one of ten health-care food fungus strains.
Because the application value of monascus pigment is far higher than that of other pigments, most of the current researches concentrate on monascus strain improvement so as to improve the yield of monascus pigment, such as the screening of space monascus FuH-23-4 in patent CN 108641968A and the application of the monascus pigment in the production of monascus pigment, and space mutagenesis is utilized to screen out the space monascus strain with higher monascus pigment production capability. In recent years, lovastatin and derivatives thereof, which are secondary metabolites produced by fermentation of monascus, are found to have the effects of reducing cholesterol, reducing blood pressure and the like, and attention is paid, but more functional strains of monascus still need to be explored.
Disclosure of Invention
Aiming at the defects or improvement demands of the prior art, the invention provides monascus purpureus CEWL and application thereof in preparing liver protection products, and aims to find a novel strain of monascus purpureus (monascus purpureus CEWL) which is preserved in China Center for Type Culture Collection (CCTCC) NO: M20231512, and experiments show that the activity of acetaldehyde dehydrogenase of the monascus purpureus CEWL strain is higher than 4U/mL, can obviously improve chemical liver injury, has good anti-alcohol and liver protection effects, and can be used for preparing liver protection products, thereby solving the technical problems of single function and narrow application range of the conventional monascus purpureus.
In order to achieve the above object, according to one aspect of the present invention, there is provided an aspergillus rhodochrous strain CEWL, designated Monascus purpureus CESWGF001, which has been deposited in the China center for type culture collection (cctccc No. M20231512).
Preferably, the activity of the acetaldehyde dehydrogenase produced by the monascus purpureus strain CEWL is above 4U/mL.
According to another aspect of the invention there is also provided the use of a strain CEWL of A.rhodochrous as described herein in the preparation of a liver protection product.
Preferably, the monascus purpureus strain CEWL is applied to preparing liver protection products, and the monascus purpureus strain CEWL is applied to preparing medicines for preventing or improving liver injury.
Preferably, the monascus purpureus strain CEWL is applied to preparing liver protection products, and the application is applied to preparing medicines for preventing alcoholic liver injury.
Preferably, the monascus purpureus strain CEWL is applied to preparing liver protection products, and the application is applied to preparing medicines for improving chemical liver injury.
In addition, the invention also provides a medicine for preventing or improving liver injury, and the active ingredients of the medicine comprise the monascus purpureus strain CEWL.
Preferably, the number of viable bacteria of the monascus purpureus strain CEWL is more than 3×10 7 CFU/mL.
In general, compared with the prior art, the above technical solution contemplated by the present invention can obtain the following beneficial effects due to the discovery that the aspergillus purpureus strain CEWL has a remarkable liver protection effect:
The monascus purpureus strain CEWL provided by the invention can reduce the activity of ALT and AST by being subjected to the verification of a liver injury model, and has better effect of reducing the activity of aminotransferase compared with the existing liver protection medicine bifendate; HE staining shows that the damaged liver cells are obviously reduced, the improvement effect of the HE staining on the liver damage caused by carbon tetrachloride is equivalent to that of the existing liver-protecting medicine bifendate, and the HE staining can be used for preparing liver-protecting products.
Drawings
FIG. 1 is the results of ALT activity assays;
FIG. 2 is an AST activity assay result;
FIG. 3 shows the HE staining results.
Detailed Description
The present invention will be described in further detail with reference to the following examples in order to make the objects, technical solutions and advantages of the present invention more apparent. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the invention. In addition, the technical features of the embodiments of the present invention described below may be combined with each other as long as they do not collide with each other.
According to the invention, monascus is screened from distiller's yeast, and a strain CEWL with an acetaldehyde dehydrogenase activity as high as 4.18U/mL is unexpectedly discovered, which has obvious effects of degrading alcohol and protecting liver, and the strain is identified as monascus purpureus, is named Monascus purpureus CESWGF001 and is preserved in China center for type culture collection, and the preservation number is CCTCC NO: M20231512.
The invention provides a monascus purpureus strain CEWL which is preserved in China Center for Type Culture Collection (CCTCC) in Wuhan City in 2023, 8 and 21 days, wherein the preservation number is CCTCC NO: M20231512 and is named as Monascus purpureus CESWGF001.
The activity of acetaldehyde dehydrogenase produced by the monascus purpureus strain CEWL is more than 4U/mL.
Liver injury is the destruction of liver cells caused by a variety of causes, including viral hepatitis, fatty liver disease, alcoholic liver disease, drug-induced liver injury, and autoimmune liver disease, thereby affecting a range of functions of the liver such as digestion, detoxification, metabolism, synthesis, etc., wherein alcoholism is likely to cause alcoholic liver injury.
Alcohol is catalyzed in the stomach by Alcohol Dehydrogenase (ADH) on its gastric mucosa to oxidize ethanol to acetaldehyde, which is then converted to acetic acid by acetaldehyde dehydrogenase (ALDH). Therefore, the oxidative metabolism of the ethanol in the stomach is quickened, the biological utilization of the ethanol by an organism is reduced, the toxic action of the ethanol on the liver can be reduced, the activity of the acetaldehyde dehydrogenase of the monascus purpureus strain CEWL is higher and can reach 4.18U/mL, the oxidative metabolism of the ethanol in the stomach can be promoted, the damage of the ethanol on the liver can be reduced, and the monascus purpureus strain CEWL can be used for preventing alcoholic liver damage.
In addition, the invention also provides an application of the monascus purpureus strain CEWL to the preparation of liver protection products.
The application is applied to the preparation of medicines for preventing or improving liver injury.
Preferably applied to the preparation of medicines for preventing alcoholic liver injury.
Preferably used for preparing medicines for improving chemical liver injury.
A medicine for preventing or improving liver injury comprises Aspergillus purpureus strain CEWL as effective component.
The number of viable bacteria of the monascus purpureus strain CEWL is more than 3 multiplied by 10 7 CFU/mL;
In some embodiments the A.rhodochrous strain CEWL is present in an amount of 10 9 CFU/mL.
The following are examples:
EXAMPLE 1 isolation of Aspergillus purpureus CEWL Strain 18
Sample: the distiller's yeast is used for separating strains by adopting the distiller's yeast used for brewing wine in a certain winery, which shows that the safety and genetic stability of the strains in the distiller's yeast are practically verified, and the screened strains can be directly used in other fields.
Potato culture medium: 200g of potato, 20g of glucose, 1L of purified water and natural pH.
Specific implementation steps
A dilution of the sample: grinding distiller's yeast with a mortar, weighing 2g, mixing with 10ml phosphoric acid buffer solution, diluting to obtain diluent, coating the diluent on potato culture medium plate, and anaerobic culturing at 30deg.C for 5d.
B single colony purification: selecting single colony according to the shape, color and size of colony, dividing the single colony on a potato culture medium plate, selecting edge hypha of the single colony for purification for multiple times to obtain pure bacteria, scraping the colony for 16s identification and seed preservation, identifying that the strain belongs to monascus purpureus, and naming the strain as Monascus purpureus CESWGF < 001 >.
The separated new strain of monascus purpureus is sent to China center for type culture collection (China center for type culture Collection), and the collection number is CCTCC NO: M20231512 at 2023, 8 and 21.
Example 2 action of Monascus purpureus CEWL Strain on degrading alcohol
(1) Degradation of ethanol
Taking out Aspergillus purpureus CEWL strain, and placing in a liquid culture medium constant temperature incubator at 37 ℃ for culturing 24h according to 10% of inoculation amount to obtain 1-generation bacterial suspension. The 1-generation bacterial suspension is inoculated into MRS broth culture medium again according to the inoculation amount of 10 percent, and is cultivated at the constant temperature of 37 ℃ for 18 h to obtain the 2-generation bacterial suspension. The activated strain is inoculated into a culture solution containing 10 percent, 15 percent and 24 percent of ethanol according to 5 percent, evenly mixed and stood, and anaerobic culture is carried out at 37 ℃ for 24 h. The ethanol residue was measured by potassium dichromate-sulfuric acid method using MRS ethanol culture solution without bacterial solution as a control, and the ethanol degradation ability of the strain was analyzed, and the results were as follows:
The degradation rates of the Aspergillus purpureus CEWL strain on 10%, 15% and 24% ethanol were 32.44%, 14.63% and 5.9%, respectively. The strain has good anti-alcoholic effect and is beneficial to preventing alcoholic liver injury.
(2) ADH and ALDH Activity assays
Activating Aspergillus purpureus CEWL strain, taking out the strain from a refrigerator at-80 ℃, inoculating the strain into a potato solid culture medium constant temperature incubator at 30 ℃ for culturing for 4-7d, adding 5mL of physiological saline into a potato culture medium flat plate by using PBS, washing and collecting spores to prepare 10 6 CFUm/L spore fungus suspension, and inoculating the strain into a potato liquid culture medium at 30 ℃ at 200rpm for culturing 48 h according to 5%.
The fungi were isolated by centrifugation at 4100 rpm for 10 minutes, and assayed using the Alcohol Dehydrogenase (ADH) assay kit and the aldehyde dehydrogenase (ALDH) assay kit, resulting in an ALDH activity of up to 4.18U/mL in A.rhodochrous CEWL strain.
First pass metabolism (FIRSTPASS METABOLISM, FPM) of ethanol refers to the phenomenon in which a portion of ethanol is oxidatively metabolized in the stomach after drinking. The metabolic process of ethanol in vivo is specifically as follows: alcohol is catalyzed in the stomach by Alcohol Dehydrogenase (ADH) on its gastric mucosa to oxidize ethanol to acetaldehyde, which is then converted to acetic acid by acetaldehyde dehydrogenase (ALDH). Therefore, the oxidative metabolism of the ethanol in the stomach is quickened, the biological utilization of the ethanol by the organism is reduced, and the toxic action of the ethanol on organs such as liver, brain and the like can be reduced.
The aspergillus purpureus CEWL provided by the invention can produce ALDH and has high ALDH activity, good anti-alcoholic effect, promotion of oxidative metabolism of ethanol by an organism, and contribution to protecting organs such as liver, brain and the like, and can be used for preventing alcoholic liver injury.
EXAMPLE 3 liver protection effect of Aspergillus purpureus CEWL Strain
Experimental materials:
mice: C57B male mice, 8 weeks old, weight 20-25 g;
strains: monascus purpureus CEWL18 and 18.
The experimental procedure is as follows:
(1) Grouping treatment for establishing mouse liver injury model
Healthy C57B male mice are selected for 32, and the mice are subjected to adaptive feeding for one week before the experiment, so that the mice are familiar with the environment, and the original self-rhythms of the mice are restored. The mice were randomly divided into 4 groups according to initial body weight, respectively a normal control group, a liver injury group (CCl 4 group), a liver injury and administration of a biphenyldicarboxylate group (ccl+biphenyldicarboxylate), a liver injury gastric lavage probiotic group (ccl+ CEWL 18); each group of mice was specifically treated as follows:
① Normal control group, free drinking water, mice were infused with normal saline on days 1-12, 3 times a week, and the mice were given intraperitoneal injections of olive oil for 12 days.
② Liver injury model group mice were perfused with normal saline on days 1-12 with free drinking water, 3 times a week, and injected intraperitoneally with 10% carbon tetrachloride (CCl 4) for 12 days.
③ Positive control group (ccl4+biphenyldicarboxylate) free drinking water, mice were perfused with biphenyldicarboxylate (2.9 mg/kg) on days 1-12, 3 times per week, and 10% carbon tetrachloride (CCL 4) was intraperitoneally injected for 12 days.
④ The strain experimental group (CCL4+ CEWL 18) was free drinking water, and mice were perfused with a spore suspension of the strain (3X 10 7 CFU/kg) on days 1-12, 3 times per week, and 10% carbon tetrachloride (CCl 4) was intraperitoneally injected into the mice for 12 days, at a dose ratio of 10:1 between mice and adults, which in this example corresponds to CEWL18 administered 3X 10 6 CFU/kg to adults.
Carbon tetrachloride and olive oil are mixed according to the proportion of 1:9 to prepare a carbon tetrachloride solution with the concentration of 10%, and the mice are subjected to gastric lavage according to the dosage of 5mL/kg by adopting an intraperitoneal injection mode. Normal control mice were injected with olive oil at the same dose.
(2) Sampling detection
Fasted 24 hours after gastric lavage, mouse serum is collected, mice are dissected, and livers are taken for detection, specifically as follows:
(2-1) detection of glutamic-oxaloacetic transaminase (AST) and glutamic-pyruvic transaminase (ALT) Activity
Collecting a mouse serum sample: after the administration time is over, the mice in each group are subjected to eyeball blood taking respectively after 24 hours of fasted, and are respectively put into marked clean EP tubes, and are placed at 4 ℃ to be used for detecting serum aminotransferase (ALT and AST);
Sample detection: taking out the eyeball blood of the taken mice; centrifuging at 3000rpm at 4deg.C for 30min; after centrifugation, the supernatant was taken into a new 1.5ml of EP, and the samples were tested using an aspartate aminotransferase (glutamic oxaloacetic transaminase/AST/GOT) test box and an alanine aminotransferase (glutamic pyruvic transaminase/ALT/GPT) test box purchased by Nanjing institute of biological engineering, and the results are shown in FIG. 1 and FIG. 2, wherein FIG. 1 shows ALT activity test results and FIG. 2 shows AST activity test results.
Glutamic-pyruvic transaminase mainly exists in liver cells, and glutamic-pyruvic transaminase mainly exists in heart cells, wherein the content of glutamic-pyruvic transaminase in liver cells is 100 times that of glutamic-pyruvic transaminase in blood. Under normal conditions, the ALT and AST contents in serum are very low, and the normal values are all in the range of 0-40U/L. Generally, the two enzymes are raised simultaneously, wherein when 1% of liver cells are damaged, glutamic pyruvic transaminase can be raised twice, and currently, glutamic pyruvic transaminase is the most sensitive index for reflecting liver damage. Clinically, glutamic-pyruvic transaminase and glutamic-oxaloacetic transaminase are together generally referred to as transaminase, and are used for assessing liver injury and function.
As can be seen from fig. 1 and 2, the ALT activity and AST activity of the mice in the normal control group are lower than 20U/L, while the ALT activity and AST activity of the mice in the liver injury model group are higher than 40U/L, compared with the mice in the normal control group, the ALT activity and AST activity of the mice in the liver injury model group are obviously increased, and are higher than 40U/L, which indicates that the modeling of the liver injury model is successful; and the ALT activity and AST activity of the mice with the liver injury model are obviously reduced by the strain of the gastric lavage bifendate or CEWL, and the ALT activity and AST activity of the mice with the strain of the gastric lavage CEWL are reduced to a lower degree, which shows that compared with the existing liver protection medicament bifendate, the strain CEWL has better liver protection effect.
(2-2) HE staining detection of liver tissue damage
Collecting a liver tissue sample: after the administration time is over, the mice are sacrificed by cervical dislocation, the abdominal cavity is cut off rapidly and carefully, the liver tissue is removed and carefully cleaned by normal saline, and small liver tissue blocks with the size of about 2.0cm multiplied by 1cm are cut and put into formalin solution for soaking for HE staining. After the operation is finished, the mouse carcasses are uniformly recovered and treated.
Sample detection: the HE staining is adopted to detect the damage condition of liver tissues, and the experimental method is briefly as follows:
(1) Preparing paraffin sections from the obtained liver tissue specimen to be tested (the cut materials should be small and thin so as to facilitate the fixative to quickly penetrate into the interior, the thickness is generally not more than 2mm, and the size is not more than 5X 5mm 2.);
(2) Putting paraffin sections into dimethylbenzene for soaking dewaxing for 5-10min;
(3) Soaking paraffin section in new xylene, dewaxing for 5-10min;
(4) Sequentially placing the slices into absolute ethanol for 5min, ethanol with 90% of 2min, ethanol with 80% of 2min and ethanol with 70% of 2min, and finally washing with distilled water for 2min;
(5) Placing the sections into hematoxylin staining solution for staining for 5-10min (the staining time is determined according to the staining result and the requirements);
(6) Immersing the slice in tap water to remove redundant staining solution for about 10min;
(7) Putting the slices into new distilled water again, and cleaning again (for a few seconds); placing the slice into eosin staining solution for staining for 30 seconds to 2 minutes (the time length is determined according to the staining result and the requirement);
(8) The slices are put into 70% ethanol for 2 times of washing;
(9) Sequentially placing the slices into 70% ethanol for 10s,80% ethanol for 10s,90% ethanol for 10s, and absolute ethanol for 10s, and finally, transparent with xylene for 5min (dehydrated ethanol concentration is from low to high in order to avoid strong tissue shrinkage caused by severe diffusion);
(10) Putting the slices into fresh dimethylbenzene again, and then, carrying out transparency for 5min; .
(11) Placing the slice at a ventilation place and sealing the slice by adopting neutral resin;
(12) Observing under an inverted microscope, taking a picture, storing and analyzing data, wherein the cell nucleus is blue and the cell plasma is pink or red; the results are shown in FIG. 3.
As shown in fig. 3, compared with the normal control group, the liver tissue injury of the mice in the liver injury model group is obvious, and the liver tissue injury of the mice in the liver injury model group is obviously recovered by the existing liver protection drugs of bifendate and CEWL strain, which indicates that the CEWL strain has obvious liver protection effect, the effect of which can be compared favorably with the effect of the existing liver protection drugs of bifendate, even better, and can be applied to the preparation of liver protection drugs or probiotic preparations.
It will be readily appreciated by those skilled in the art that the foregoing description is merely a preferred embodiment of the invention and is not intended to limit the invention, but any modifications, equivalents, improvements or alternatives falling within the spirit and principles of the invention are intended to be included within the scope of the invention.
Claims (3)
1. A monascus purpureus strain CEWL is characterized in that the strain is named Monascus purpureus CESWGF001 and is preserved in China Center for Type Culture Collection (CCTCC) with the preservation number of M20231512.
2. The aspergillus rhodochrous strain CEWL as claimed in claim 1, which produces an acetaldehyde dehydrogenase activity of more than 4U/mL.
3. Use of the strain CEWL of monascus purpureus 18 according to claim 1 or 2 for the preparation of a liver protection product, wherein said liver protection product is a medicament for preventing alcoholic liver injury or improving chemical liver injury.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311431255.9A CN117448178B (en) | 2023-10-31 | 2023-10-31 | Monascus purpureus CEWL and application thereof in preparation of liver protection products |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311431255.9A CN117448178B (en) | 2023-10-31 | 2023-10-31 | Monascus purpureus CEWL and application thereof in preparation of liver protection products |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN117448178A CN117448178A (en) | 2024-01-26 |
| CN117448178B true CN117448178B (en) | 2024-04-26 |
Family
ID=89579551
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311431255.9A Active CN117448178B (en) | 2023-10-31 | 2023-10-31 | Monascus purpureus CEWL and application thereof in preparation of liver protection products |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117448178B (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1686519A (en) * | 2005-03-23 | 2005-10-26 | 张帆 | Medicinal composition for preventing alcoholic liver damage |
| CN101073602B (en) * | 2006-05-15 | 2011-05-11 | 成都地奥九泓制药厂 | Medicinal composition for treating or improving liver function, its production and use |
| TW201313897A (en) * | 2011-09-27 | 2013-04-01 | Tzu Hai Tung Food Co Ltd | Method of producing antioxidant active substance from agricultural byproduct |
| CN109554301A (en) * | 2018-11-22 | 2019-04-02 | 浙江三禾生物工程股份有限公司 | A kind of monascus purpureus, fermentation process and tunning reduce the application in interior fat product in preparation |
| CN110016440A (en) * | 2019-05-15 | 2019-07-16 | 四川剑南春（集团）有限责任公司 | Monascus purpureus and its screening technique and purposes |
| CN114668783A (en) * | 2022-05-30 | 2022-06-28 | 微康益生菌(苏州)股份有限公司 | Application of lactobacillus plantarum strain WKA86 in preparing liver protection/antioxidation/prevention and treatment of alcoholic liver injury medicines |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111423986B (en) * | 2020-01-19 | 2022-03-25 | 北京工商大学 | Monascus purpureus strain capable of producing esterase at high yield and application of monascus purpureus strain in preparation of ester-flavor monascus cheese |
-
2023
- 2023-10-31 CN CN202311431255.9A patent/CN117448178B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1686519A (en) * | 2005-03-23 | 2005-10-26 | 张帆 | Medicinal composition for preventing alcoholic liver damage |
| CN101073602B (en) * | 2006-05-15 | 2011-05-11 | 成都地奥九泓制药厂 | Medicinal composition for treating or improving liver function, its production and use |
| TW201313897A (en) * | 2011-09-27 | 2013-04-01 | Tzu Hai Tung Food Co Ltd | Method of producing antioxidant active substance from agricultural byproduct |
| CN109554301A (en) * | 2018-11-22 | 2019-04-02 | 浙江三禾生物工程股份有限公司 | A kind of monascus purpureus, fermentation process and tunning reduce the application in interior fat product in preparation |
| CN110016440A (en) * | 2019-05-15 | 2019-07-16 | 四川剑南春（集团）有限责任公司 | Monascus purpureus and its screening technique and purposes |
| CN114668783A (en) * | 2022-05-30 | 2022-06-28 | 微康益生菌(苏州)股份有限公司 | Application of lactobacillus plantarum strain WKA86 in preparing liver protection/antioxidation/prevention and treatment of alcoholic liver injury medicines |
Non-Patent Citations (3)
| Title |
|---|
| Protective Effect of Monascus-Fermented Red Mold Rice against Alcoholic Liver Disease by Attenuating Oxidative Stress and Inflammatory Response;Chin-fu Cheng等;《Journal of Agricutural and Food Chemistry》;20110823(第59期);第9950-9957页 * |
| 传统白酒发酵过程中微生物种类及功能研究进展;赵龙飞等;《食品研究与开发》;20171010(第19期);第205-208页 * |
| 红曲改善大鼠非酒精性脂肪肝的组织与生化学研究;罗文政等;《中华全科医学》;20111208;第9卷(第12期);第1842-1853页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN117448178A (en) | 2024-01-26 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN115322911A (en) | Abnormal yeast Weikehan and application thereof | |
| CN112852780A (en) | Monascus flavus and application thereof in preparing tannase and degrading tannin | |
| CN117448178B (en) | Monascus purpureus CEWL and application thereof in preparation of liver protection products | |
| CN116355816A (en) | Microorganism of fermented samara oil seed and blood lipid reducing composition thereof | |
| CN114456990B (en) | Preparation method of DDGS and fermentation strain and culture medium thereof | |
| CN116426431A (en) | Bacillus natto and application thereof | |
| CN114854623B (en) | Lactobacillus delbrueckii subspecies bulgaricus, microbial inoculum containing lactobacillus delbrueckii subspecies bulgaricus and application of lactobacillus delbrueckii subspecies bulgaricus | |
| CN114533768B (en) | Application of lactobacillus probiotics CGMCC No.1.13855 in preparing medicament for treating diabetes | |
| CN115992059A (en) | Lactobacillus johnsonii for producing feruloyl esterase and application thereof in relieving ulcerative colitis | |
| CN115895977A (en) | Lactobacillus paracasei and application thereof | |
| CN112442453B (en) | Ormokodak yeast for degrading biogenic amine and application of same in white spirit brewing | |
| CN111676160B (en) | Application of beautiful millettia root endophyte RH5 in promoting strong growth of beautiful millettia root | |
| CN110358702B (en) | Serratia and application thereof in degradation of vomitoxin | |
| CN117210337B (en) | Oospore fungus with chicken feather degradation activity and application thereof | |
| CN113667623B (en) | Bacillus belgii preparation for degrading acetaldehyde and preparation method and application thereof | |
| CN114149953B (en) | Staphylococcus gallinarum and application thereof in removing volatile amine in aquatic products | |
| CN117363524B (en) | Lactobacillus gasseri MY4 and application thereof in preparation of sleep-aiding and whitening medicines | |
| CN117683697B (en) | Bacillus bailii Y01 and application thereof in bacteriostasis and improvement of animal growth performance | |
| CN113046275B (en) | Bacillus used for prawn feed and fermented Chinese herbal medicine preparation used for prawn feed | |
| CN114854611B (en) | Selenium-enriched saccharomycetes, probiotics composite bacterial liquid and fermented soybean meal feed | |
| CN116849291B (en) | Fermented feed for enhancing animal immunity and preparation method thereof | |
| Ingledew et al. | THE FATE OF LIVE BREWERS'YEAST SLURRY IN BOVINE RUMEN FLUID | |
| CN114591863B (en) | Tibetan pig source fungicide combination for improving growth performance and immune function of weaned piglets | |
| CN101215525B (en) | Method for separating microorganism capable of producing ferulic acid esterase | |
| CN117363492A (en) | Composite strain for degrading tea residue insoluble dietary fibers and application thereof |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |