CN113881750A - A method for preparing ginsenoside preparation by biological engineering technology - Google Patents
A method for preparing ginsenoside preparation by biological engineering technology Download PDFInfo
- Publication number
- CN113881750A CN113881750A CN202111224254.8A CN202111224254A CN113881750A CN 113881750 A CN113881750 A CN 113881750A CN 202111224254 A CN202111224254 A CN 202111224254A CN 113881750 A CN113881750 A CN 113881750A
- Authority
- CN
- China
- Prior art keywords
- preparation
- fermentation
- lactobacillus
- ginsenoside
- culture medium
- 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.)
- Withdrawn
Links
- 238000002360 preparation method Methods 0.000 title claims abstract description 130
- 229930182494 ginsenoside Natural products 0.000 title claims abstract description 46
- 229940089161 ginsenoside Drugs 0.000 title claims abstract description 44
- 238000000034 method Methods 0.000 title claims abstract description 25
- 238000005516 engineering process Methods 0.000 title claims abstract description 16
- 102000004190 Enzymes Human genes 0.000 claims abstract description 53
- 108090000790 Enzymes Proteins 0.000 claims abstract description 53
- 229930182490 saponin Natural products 0.000 claims abstract description 53
- 150000007949 saponins Chemical class 0.000 claims abstract description 53
- 239000001397 quillaja saponaria molina bark Substances 0.000 claims abstract description 48
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims abstract description 28
- 235000020357 syrup Nutrition 0.000 claims abstract description 26
- 239000006188 syrup Substances 0.000 claims abstract description 26
- 238000010438 heat treatment Methods 0.000 claims abstract description 25
- 150000001875 compounds Chemical class 0.000 claims abstract description 16
- 235000014655 lactic acid Nutrition 0.000 claims abstract description 14
- 239000004310 lactic acid Substances 0.000 claims abstract description 14
- 150000002772 monosaccharides Chemical class 0.000 claims abstract description 9
- 230000004151 fermentation Effects 0.000 claims description 76
- 238000000855 fermentation Methods 0.000 claims description 73
- 238000004321 preservation Methods 0.000 claims description 42
- 239000001963 growth medium Substances 0.000 claims description 38
- 239000000047 product Substances 0.000 claims description 30
- 241000894006 Bacteria Species 0.000 claims description 26
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 23
- 239000000203 mixture Substances 0.000 claims description 21
- 241000186660 Lactobacillus Species 0.000 claims description 20
- 229940039696 lactobacillus Drugs 0.000 claims description 20
- 239000007787 solid Substances 0.000 claims description 16
- 239000007788 liquid Substances 0.000 claims description 15
- 238000003756 stirring Methods 0.000 claims description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 15
- 241000533950 Leucojum Species 0.000 claims description 11
- 238000002156 mixing Methods 0.000 claims description 11
- 239000000843 powder Substances 0.000 claims description 9
- 238000010563 solid-state fermentation Methods 0.000 claims description 9
- 238000001514 detection method Methods 0.000 claims description 8
- 241000186679 Lactobacillus buchneri Species 0.000 claims description 7
- 241000186605 Lactobacillus paracasei Species 0.000 claims description 7
- 240000006024 Lactobacillus plantarum Species 0.000 claims description 7
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims description 7
- 241000577554 Lactobacillus zeae Species 0.000 claims description 7
- 241000509461 [Candida] ethanolica Species 0.000 claims description 7
- 229940072205 lactobacillus plantarum Drugs 0.000 claims description 7
- 239000006228 supernatant Substances 0.000 claims description 7
- 244000061456 Solanum tuberosum Species 0.000 claims description 6
- 235000002595 Solanum tuberosum Nutrition 0.000 claims description 6
- 230000004913 activation Effects 0.000 claims description 6
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 6
- 230000001580 bacterial effect Effects 0.000 claims description 6
- 238000001816 cooling Methods 0.000 claims description 6
- 238000001704 evaporation Methods 0.000 claims description 6
- 235000019534 high fructose corn syrup Nutrition 0.000 claims description 6
- 239000000463 material Substances 0.000 claims description 6
- 229910052760 oxygen Inorganic materials 0.000 claims description 6
- 239000001301 oxygen Substances 0.000 claims description 6
- 239000008223 sterile water Substances 0.000 claims description 6
- 239000000126 substance Substances 0.000 claims description 6
- 240000007594 Oryza sativa Species 0.000 claims description 5
- 235000007164 Oryza sativa Nutrition 0.000 claims description 5
- 241000209140 Triticum Species 0.000 claims description 5
- 235000021307 Triticum Nutrition 0.000 claims description 5
- 230000008020 evaporation Effects 0.000 claims description 5
- 235000013575 mashed potatoes Nutrition 0.000 claims description 5
- 235000009566 rice Nutrition 0.000 claims description 5
- 238000010025 steaming Methods 0.000 claims description 4
- KEQXNNJHMWSZHK-UHFFFAOYSA-L 1,3,2,4$l^{2}-dioxathiaplumbetane 2,2-dioxide Chemical compound [Pb+2].[O-]S([O-])(=O)=O KEQXNNJHMWSZHK-UHFFFAOYSA-L 0.000 claims description 3
- FFRBMBIXVSCUFS-UHFFFAOYSA-N 2,4-dinitro-1-naphthol Chemical group C1=CC=C2C(O)=C([N+]([O-])=O)C=C([N+]([O-])=O)C2=C1 FFRBMBIXVSCUFS-UHFFFAOYSA-N 0.000 claims description 3
- 230000003213 activating effect Effects 0.000 claims description 3
- 238000010564 aerobic fermentation Methods 0.000 claims description 3
- 239000003205 fragrance Substances 0.000 claims description 3
- 238000011081 inoculation Methods 0.000 claims description 3
- 230000002503 metabolic effect Effects 0.000 claims description 3
- 229920006395 saturated elastomer Polymers 0.000 claims description 3
- 238000007789 sealing Methods 0.000 claims description 3
- 239000000758 substrate Substances 0.000 claims description 3
- 230000000007 visual effect Effects 0.000 claims description 3
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 claims description 2
- DBTMGCOVALSLOR-UHFFFAOYSA-N 32-alpha-galactosyl-3-alpha-galactosyl-galactose Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(OC2C(C(CO)OC(O)C2O)O)OC(CO)C1O DBTMGCOVALSLOR-UHFFFAOYSA-N 0.000 claims description 2
- RXVWSYJTUUKTEA-UHFFFAOYSA-N D-maltotriose Natural products OC1C(O)C(OC(C(O)CO)C(O)C(O)C=O)OC(CO)C1OC1C(O)C(O)C(O)C(CO)O1 RXVWSYJTUUKTEA-UHFFFAOYSA-N 0.000 claims description 2
- 229930091371 Fructose Natural products 0.000 claims description 2
- 239000005715 Fructose Substances 0.000 claims description 2
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 claims description 2
- 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 claims description 2
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 claims description 2
- 241000235342 Saccharomycetes Species 0.000 claims description 2
- 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 claims description 2
- 229930006000 Sucrose Natural products 0.000 claims description 2
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims description 2
- 235000013361 beverage Nutrition 0.000 claims description 2
- 239000008103 glucose Substances 0.000 claims description 2
- FYGDTMLNYKFZSV-UHFFFAOYSA-N mannotriose Natural products OC1C(O)C(O)C(CO)OC1OC1C(CO)OC(OC2C(OC(O)C(O)C2O)CO)C(O)C1O FYGDTMLNYKFZSV-UHFFFAOYSA-N 0.000 claims description 2
- 239000005720 sucrose Substances 0.000 claims description 2
- FYGDTMLNYKFZSV-BYLHFPJWSA-N β-1,4-galactotrioside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@H](CO)O[C@@H](O[C@@H]2[C@@H](O[C@@H](O)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O FYGDTMLNYKFZSV-BYLHFPJWSA-N 0.000 claims description 2
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 claims 2
- 229960002160 maltose Drugs 0.000 claims 2
- 229960002737 fructose Drugs 0.000 claims 1
- 229960001031 glucose Drugs 0.000 claims 1
- 230000008821 health effect Effects 0.000 claims 1
- 235000020374 simple syrup Nutrition 0.000 claims 1
- 229960004793 sucrose Drugs 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 5
- OYHQOLUKZRVURQ-HZJYTTRNSA-N Linoleic acid Chemical compound CCCCC\C=C/C\C=C/CCCCCCCC(O)=O OYHQOLUKZRVURQ-HZJYTTRNSA-N 0.000 abstract description 4
- 150000001408 amides Chemical class 0.000 abstract description 4
- 125000000539 amino acid group Chemical group 0.000 abstract description 4
- 239000005515 coenzyme Substances 0.000 abstract description 4
- 238000009833 condensation Methods 0.000 abstract description 4
- 230000005494 condensation Effects 0.000 abstract description 4
- OYHQOLUKZRVURQ-IXWMQOLASA-N linoleic acid Natural products CCCCC\C=C/C\C=C\CCCCCCCC(O)=O OYHQOLUKZRVURQ-IXWMQOLASA-N 0.000 abstract description 4
- 235000020778 linoleic acid Nutrition 0.000 abstract description 4
- 230000018044 dehydration Effects 0.000 abstract description 3
- 238000006297 dehydration reaction Methods 0.000 abstract description 3
- 238000006482 condensation reaction Methods 0.000 abstract description 2
- 150000002373 hemiacetals Chemical class 0.000 abstract description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 abstract description 2
- 125000002485 formyl group Chemical class [H]C(*)=O 0.000 abstract 1
- 229940088598 enzyme Drugs 0.000 description 46
- 235000017709 saponins Nutrition 0.000 description 43
- 238000006243 chemical reaction Methods 0.000 description 8
- 239000010410 layer Substances 0.000 description 6
- 230000008520 organization Effects 0.000 description 6
- 241000208340 Araliaceae Species 0.000 description 4
- 235000003140 Panax quinquefolius Nutrition 0.000 description 4
- 230000002779 inactivation Effects 0.000 description 4
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 description 3
- 150000002016 disaccharides Chemical class 0.000 description 3
- 235000008434 ginseng Nutrition 0.000 description 3
- 235000013402 health food Nutrition 0.000 description 3
- -1 steroid compound Chemical class 0.000 description 3
- PJVXUVWGSCCGHT-ZPYZYFCMSA-N (2r,3s,4r,5r)-2,3,4,5,6-pentahydroxyhexanal;(3s,4r,5r)-1,3,4,5,6-pentahydroxyhexan-2-one Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O.OC[C@@H](O)[C@@H](O)[C@H](O)C(=O)CO PJVXUVWGSCCGHT-ZPYZYFCMSA-N 0.000 description 2
- 230000032683 aging Effects 0.000 description 2
- 150000001299 aldehydes Chemical class 0.000 description 2
- 229940079919 digestives enzyme preparation Drugs 0.000 description 2
- YURJSTAIMNSZAE-HHNZYBFYSA-N ginsenoside Rg1 Chemical compound O([C@@](C)(CCC=C(C)C)[C@@H]1[C@@H]2[C@@]([C@@]3(C[C@@H]([C@H]4C(C)(C)[C@@H](O)CC[C@]4(C)[C@H]3C[C@H]2O)O[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)O)C)(C)CC1)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O YURJSTAIMNSZAE-HHNZYBFYSA-N 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 230000007062 hydrolysis Effects 0.000 description 2
- 238000006460 hydrolysis reaction Methods 0.000 description 2
- 239000011229 interlayer Substances 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 235000000346 sugar Nutrition 0.000 description 2
- FBFMBWCLBGQEBU-GYMUUCMZSA-N 20-gluco-ginsenoside-Rf Natural products O([C@](CC/C=C(\C)/C)(C)[C@@H]1[C@H]2[C@H](O)C[C@H]3[C@](C)([C@]2(C)CC1)C[C@H](O[C@@H]1[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O2)[C@@H](O)[C@H](O)[C@@H](CO)O1)[C@H]1C(C)(C)[C@@H](O)CC[C@]31C)[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 FBFMBWCLBGQEBU-GYMUUCMZSA-N 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 240000008620 Fagopyrum esculentum Species 0.000 description 1
- 235000009419 Fagopyrum esculentum Nutrition 0.000 description 1
- UZIOUZHBUYLDHW-MSJHMJQNSA-N Ginsenoside Rf Natural products O([C@H]1[C@@H](O)[C@H](O)[C@H](CO)O[C@@H]1O[C@@H]1[C@H]2C(C)(C)[C@@H](O)CC[C@]2(C)[C@@H]2[C@](C)([C@@]3(C)[C@H]([C@@H](O)C2)[C@@H]([C@@](O)(CC/C=C(\C)/C)C)CC3)C1)[C@H]1[C@@H](O)[C@@H](O)[C@H](O)[C@H](CO)O1 UZIOUZHBUYLDHW-MSJHMJQNSA-N 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 241000180649 Panax notoginseng Species 0.000 description 1
- 235000003143 Panax notoginseng Nutrition 0.000 description 1
- 240000005373 Panax quinquefolius Species 0.000 description 1
- 240000006394 Sorghum bicolor Species 0.000 description 1
- 235000011684 Sorghum saccharatum Nutrition 0.000 description 1
- 208000007536 Thrombosis Diseases 0.000 description 1
- YURJSTAIMNSZAE-UHFFFAOYSA-N UNPD89172 Natural products C1CC(C2(CC(C3C(C)(C)C(O)CCC3(C)C2CC2O)OC3C(C(O)C(O)C(CO)O3)O)C)(C)C2C1C(C)(CCC=C(C)C)OC1OC(CO)C(O)C(O)C1O YURJSTAIMNSZAE-UHFFFAOYSA-N 0.000 description 1
- DHKHKXVYLBGOIT-UHFFFAOYSA-N acetaldehyde Diethyl Acetal Natural products CCOC(C)OCC DHKHKXVYLBGOIT-UHFFFAOYSA-N 0.000 description 1
- 150000001241 acetals Chemical class 0.000 description 1
- OIPILFWXSMYKGL-UHFFFAOYSA-N acetylcholine Chemical compound CC(=O)OCC[N+](C)(C)C OIPILFWXSMYKGL-UHFFFAOYSA-N 0.000 description 1
- 229960004373 acetylcholine Drugs 0.000 description 1
- 125000002252 acyl group Chemical group 0.000 description 1
- 125000003172 aldehyde group Chemical group 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 210000003169 central nervous system Anatomy 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 230000008602 contraction Effects 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 235000021551 crystal sugar Nutrition 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 235000021433 fructose syrup Nutrition 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- UZIOUZHBUYLDHW-XUBRWZAZSA-N ginsenoside Rf Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@H]2C(C)(C)[C@@H](O)CC[C@]2(C)[C@H]2C[C@@H](O)[C@H]3[C@@]([C@@]2(C1)C)(C)CC[C@@H]3[C@@](C)(O)CCC=C(C)C)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O UZIOUZHBUYLDHW-XUBRWZAZSA-N 0.000 description 1
- CBEHEBUBNAGGKC-UHFFFAOYSA-N ginsenoside Rg1 Natural products CC(=CCCC(C)(OC1OC(CO)C(O)C(O)C1O)C2CCC3(C)C2C(O)CC4C5(C)CCC(O)C(C)(C)C5CC(OC6OC(CO)C(O)C(O)C6O)C34C)C CBEHEBUBNAGGKC-UHFFFAOYSA-N 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 235000021552 granulated sugar Nutrition 0.000 description 1
- 230000002949 hemolytic effect Effects 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 238000006709 oxidative esterification reaction Methods 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 239000011435 rock Substances 0.000 description 1
- 208000010110 spontaneous platelet aggregation Diseases 0.000 description 1
- 150000008130 triterpenoid saponins Chemical class 0.000 description 1
- 230000003313 weakening effect Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P39/00—Processes involving microorganisms of different genera in the same process, simultaneously
-
- 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
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/52—Adding ingredients
-
- 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
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
-
- 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
- C12N1/16—Yeasts; Culture media therefor
-
- 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
- C12N1/20—Bacteria; Culture media therefor
-
- 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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P33/00—Preparation of steroids
- C12P33/20—Preparation of steroids containing heterocyclic rings
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Biomedical Technology (AREA)
- Mycology (AREA)
- Medicinal Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- General Chemical & Material Sciences (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Botany (AREA)
- Molecular Biology (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a method for preparing a ginsenoside preparation by adopting a biological engineering technology, wherein the ginsenoside preparation comprises at least one of a saponin-rich preparation and a saponin-rich syrup; the invention prepares the ginsenoside preparations such as saponin-rich preparation and saponin-rich syrup by adding and utilizing the activity of hemiacetal and hydroxyl in monosaccharide molecules and performing complex aldehyde condensation reaction and heating dehydration condensation through the compound biological enzyme preparation rich in amino acid residues, amide, coenzyme, lactic acid, linoleic acid and other active groups, and provides a new idea for the development and utilization of ginsenoside.
Description
Technical Field
The invention relates to the technical field of microorganism application, in particular to a method for preparing a ginsenoside preparation by adopting a biological engineering technology.
Background
Ginsenoside (Ginsenoside) is a steroid compound, also known as triterpenoid saponin. Mainly exists in ginseng medicinal materials. Ginsenoside is considered as an active component in ginseng, and has the effects of resisting thrombus, fatigue, aging, controlling tumor, enhancing immunity and the like, wherein the ginsenoside Rg1 has the effects of quickly relieving fatigue, improving learning and memory, delaying aging, exciting central nervous system, inhibiting platelet aggregation and the like, and the ginsenoside Rf has the effects of weakening intestinal canal contraction caused by acetylcholine, having hemolytic activity and the like. However, many ginsenosides naturally exist in roots, buds and stems of araliaceae plants, such as ginseng, panax notoginseng and American ginseng, but the content of the ginsenosides in the plants is low, the extraction cost is high, and the increasing medical requirements are difficult to meet.
Disclosure of Invention
Therefore, based on the above background, the present invention provides a method for preparing ginsenoside preparations by using bioengineering technology, which prepares ginsenoside preparations such as saponin-rich preparations and saponin-rich syrups by using compound biological enzyme preparations rich in amino acid residues, amides, coenzymes, and active groups such as lactic acid and linoleic acid.
The technical scheme provided by the invention is as follows:
a method for preparing ginsenoside preparation by adopting bioengineering technology comprises the following steps:
(1) selecting a compound biological enzyme preparation, wherein the biological enzyme preparation is prepared by fermenting yeast and lactobacillus combined bacteria;
(2) directly heating the biological enzyme preparation in the step (1); or adding syrup, stirring, heating, and concentrating to obtain ginsenoside preparation.
Further, the ginsenoside preparation comprises at least one of a saponin-rich preparation and a saponin-rich syrup;
the preparation method comprises the following steps:
(1) selecting a compound biological enzyme preparation, wherein the biological enzyme preparation is prepared by fermenting yeast and lactobacillus combined bacteria;
(2) adding syrup into the biological enzyme preparation in the step (1) according to the volume ratio of 0-100% and uniformly stirring;
(3) heating and concentrating the mixture obtained in the step (2);
(4) when the evaporated amount in the mixture obtained in the step (2) reaches 1/3-2/5 of the volume of the original mixture, the saponin-rich preparation can be prepared;
(5) and (3) continuing to heat the saponin-rich preparation obtained in the step (4) until the volume of the evaporated saponin-rich preparation reaches the added amount of the syrup obtained in the step (2), and cooling the material to obtain the saponin-rich syrup.
Further, the preparation steps of the compound biological enzyme preparation are as follows:
(1) selecting yeast and lactobacillus combined bacteria as strains;
(2) activating strains: moistening potato mud snowflake powder with sterile water according to a weight ratio of 1:1, mixing the strain with the moistened potato mud snowflake powder according to a weight ratio of 1:2, and carrying out aerobic activation in a sterile environment;
(3) preparation and inoculation of the culture medium: the culture medium is prepared by using mashed potato snowflake powder, wheat germ and sticky rice as main materials, adding water into the culture medium, uniformly stirring, steaming, and then airing under an aseptic environment to a temperature below 45 ℃ to finish the preparation of the culture medium;
and (3) mixing the prepared culture medium with the activated strain obtained in the step (2) according to the weight ratio of 10: 1, uniformly mixing;
(4) adjusting the humidity of the culture medium inoculated in the step (3) to ensure that the holding balls are soaked by water and the whole culture medium keeps moist and breathable;
(5) carrying out aerobic fermentation on the culture medium in the step (4) for more than 48 hours; when the culture medium has uniform white and milky plaque visible to naked eyes, transferring the culture medium into a fermentation tank; the solid fermentation substance occupies one third of the fermentation tank, and all channels of the fermentation tank are closed, so that the culture medium fermentation substance entering the tank is continuously subjected to aerobic culture in the tank body until the oxygen in the tank body is exhausted, and the fermentation tank automatically enters an anaerobic fermentation state;
(6) when the pressure in the fermentation tank is unchanged, the solid fermentation product enters saturated fermentation, the tank is opened, the rich fermentation fragrance overflows, and the abundant mycelium is covered by the solid fermentation product under the visual observationDetecting the total viable count of yeast 6.4 × 10 on the surface of solid culture medium4-5.9×105cfu/ml; total number of lactic acid bacteria 4X 108-5.1×109cfu/ml, at which point the solid state fermentation process has reached saturation, may enter the liquid state fermentation stage;
(7) adding sterile water into the solid-state fermentation product obtained in the step (6) according to the mass ratio of 1:3, fully stirring to dilute the metabolic substrate in the solid-state fermentation process, ensuring that at least 1/5 space is reserved in the fermentation tank body, carrying out aerobic activation culture for more than 72 hours until white or milk-white bacterial plaque is densely distributed on the surface of the liquid-state fermentation product, sealing all channels of the fermentation tank, and gradually exhausting oxygen from the fermentation product to enter the anaerobic fermentation process;
(8) when the surface bacterial plaque of the liquid fermentation product in the step (7) is in a folded state, the supernatant liquid in the middle layer is golden yellow, clear and transparent, and the solid fermentation product in the bottom layer and the liquid in the middle layer are distinct and are not adhered, namely the solid fermentation-liquid fermentation process is completely finished; the obtained fermentation product can be the prepared biological enzyme preparation.
Further, the preparation steps of the biological enzyme preparation also comprise:
(9) and (3) gradually heating the solution obtained in the step (8) to 78-88 ℃, cooling the solution for 2 hours, and extracting supernatant of the fermentation product to obtain the inactivated biological enzyme preparation.
Further, the yeast and fermentation combined bacteria in the preparation step 1) of the biological preparation comprise yeasts and lactic acid bacteria, wherein the yeasts comprise Candida ethanolica B-JJ1, and the lactic acid bacteria comprise at least one of Lactobacillus buchneri B-JR1, Lactobacillus paracasei B-JR2, Lactobacillus zeae B-JR4, Lactobacillus plantarum B-JR5 and lactic acid bacteria B-JR 6; the preservation number of the Candida ethanolica B-JJ1 is CCTCC NO: M2020136; the preservation number of the lactobacillus buchneri B-JR1 is CCTCC NO: M2021132; the preservation number of the lactobacillus paracasei B-JR2 is CCTCC NO: 2021133; the preservation number of the lactobacillus zeae B-JR4 is CCTCC NO: 2021135; the preservation number of the lactobacillus plantarum B-JR5 is CCTCC NO: 2021501; the preservation number of the lactobacillus B-JR6 is CCTCC NO: M20211502.
Further, the syrup is prepared from monosaccharide.
Further, in the step 2) of the ginsenoside preparation, the syrup is high fructose corn syrup.
Or further, the syrup is a syrup prepared by at least one of glucose, sucrose, fructose, maltose disaccharide and maltotriose.
The invention also provides the application of the ginsenoside preparation in preparing the beverage with the health care function.
The preservation number of the Candida ethanolica B-JJ1 is CCTCC NO: M2020136; the preservation date is 2021, 01 month and 21 days; the preservation organization is China center for type culture Collection; the preservation place is Wuhan university in China.
The preservation number of the lactobacillus buchneri B-JR1 is CCTCC NO: M2021132; the preservation date is 2021, 01 month and 21 days; the preservation organization is China center for type culture Collection; the preservation place is Wuhan university in China.
The preservation number of the lactobacillus paracasei B-JR2 is CCTCC NO: 2021133; the preservation date is 2021, 01 month and 21 days; the preservation organization is China center for type culture Collection; the preservation place is Wuhan university in China.
The preservation number of the lactobacillus zeae B-JR4 is CCTCC NO: 2021135; the preservation date is 2021, 01 month and 21 days; the preservation organization is China center for type culture Collection; the preservation place is Wuhan university in China.
The preservation number of the lactobacillus plantarum B-JR5 is CCTCC NO: 2021501; the preservation date is 2021, 05 and 07 months; the preservation organization is China center for type culture Collection; the preservation place is Wuhan university in China.
The preservation number of the lactobacillus B-JR6 is CCTCC NO: M20211502; the preservation date is 2021, 05 and 07 months; the preservation organization is China center for type culture Collection; the preservation place is Wuhan university in China.
By adopting the technical scheme, the method has the following beneficial effects:
the invention prepares the ginsenoside preparation such as saponin-rich preparation and saponin-rich syrup by adding and utilizing the activity of hemiacetal and hydroxyl in monosaccharide molecules and performing complex aldehyde condensation reaction and heating dehydration condensation through the compound biological enzyme preparation rich in amino acid residues, amide, coenzyme, lactic acid, linoleic acid and other active groups, and provides a new direction and thought for obtaining ginsenoside and products thereof.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are some embodiments of the present invention, but not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The present invention will be further described with reference to the following examples.
Example 1: a method for preparing ginsenoside preparation by biological engineering technology comprises preparing ginsenoside preparation rich in saponin;
which comprises the following steps:
(1) selecting a composite biological enzyme preparation, wherein the biological enzyme preparation is prepared by fermenting yeast and lactic acid combined bacteria;
(2) and (2) heating the compound biological enzyme preparation obtained in the step (1) to 160 ℃ until the volume of the evaporated compound biological enzyme preparation is 1/3 of the volume of the raw material, and thus obtaining the saponin-rich preparation.
The total ginsenoside content of the saponin-rich preparation obtained in step (2) of this example was determined to be 3700 mg/L.
The detection is completed by the analysis and detection center of microorganisms in Guangdong province according to the second part of the health food efficacy component and health index detection standard (twenty-three, determination of total saponins in health food) P306-307 in the health food detection and evaluation technical standard (2003 edition), and the detection of the content of the total saponins is completed accordingly.
The preparation procedure of the bio-enzyme preparation of this example is as follows:
(1) selecting yeast and lactobacillus combined bacteria as strains;
the yeast and lactic acid bacteria combined bacteria used in the present example include yeast including Candida ethanolica B-JJ1, and lactic acid bacteria including Lactobacillus buchneri B-JR1, Lactobacillus paracasei B-JR2, Lactobacillus zeae B-JR4, Lactobacillus plantarum B-JR5, and lactic acid bacteria B-JR 6; the preservation number of the Candida ethanolica B-JJ1 is CCTCC NO: M2020136; the preservation number of the lactobacillus buchneri B-JR1 is CCTCC NO: M2021132; the preservation number of the lactobacillus paracasei B-JR2 is CCTCC NO: 2021133; the preservation number of the lactobacillus zeae B-JR4 is CCTCC NO: 2021135; the preservation number of the lactobacillus plantarum B-JR5 is CCTCC NO: 2021501; the preservation number of the lactobacillus B-JR6 is CCTCC NO: M20211502.
The specific process of culturing the yeast and lactobacillus combined bacteria used in the invention can refer to the invention patent with the patent number of 202111198135X.
(2) Activating strains: moistening potato mud snowflake powder with sterile water according to a weight ratio of 1:1, mixing the strain with the moistened potato mud snowflake powder according to a weight ratio of 1:2, and carrying out aerobic activation in a sterile environment;
(3) preparation and inoculation of the culture medium: the culture medium is prepared by using mashed potato snowflake powder, wheat germ and sticky rice as main materials, adding water into the culture medium, uniformly stirring, steaming, and then airing under an aseptic environment to a temperature below 45 ℃ to finish the preparation of the culture medium;
and (3) mixing the prepared culture medium with the activated strain obtained in the step (2) according to the weight ratio of 10: 1, uniformly mixing;
the preparation process of the culture medium in the step is as follows:
mixing sorghum rice, buckwheat and glutinous rice according to the weight ratio of 50: 10: after being cleaned in a ratio of 20 percent, the wheat germ and the mashed potato snowflakes are uniformly mixed with 10 percent of wheat germ and 10 percent of mashed potato snowflakes, wherein the ratio of 1:1, adding water, stirring uniformly, putting into a cabinet, steaming, and then airing to 45 ℃ in a sterile environment to finish the preparation of the culture medium.
(4) Adjusting the humidity of the culture medium inoculated in the step (3) to ensure that the holding balls are soaked by water and the whole culture medium keeps moist and breathable;
(5) carrying out aerobic fermentation on the culture medium in the step (4) for more than 48 hours; when the culture medium has uniform white and milky plaque visible to naked eyes, transferring the culture medium into a fermentation tank; the solid fermentation substance occupies one third of the fermentation tank, and all channels of the fermentation tank are closed, so that the culture medium fermentation substance entering the tank is continuously subjected to aerobic culture in the tank body until the oxygen in the tank body is exhausted, and the fermentation tank automatically enters an anaerobic fermentation state;
(6) when the pressure in the fermentation tank is unchanged, the solid fermentation product enters saturated fermentation, the strong fermentation fragrance overflows after the fermentation tank is opened, the surface of the solid culture medium is covered with the luxuriant mycelia by visual observation, the fermentation product is taken for detection, and the total number of the viable bacteria of the saccharomycetes is 6.4 multiplied by 104-5.9×105cfu/ml; total number of lactic acid bacteria 4X 108-5.1×109cfu/ml, at which point the solid state fermentation process has reached saturation, may enter the liquid state fermentation stage;
(7) adding sterile water into the solid-state fermentation product obtained in the step (6) according to the mass ratio of 1:3, fully stirring to dilute the metabolic substrate in the solid-state fermentation process, ensuring that at least 1/5 space is reserved in the fermentation tank body, carrying out aerobic activation culture for more than 72 hours until white or milk-white bacterial plaque is densely distributed on the surface of the liquid-state fermentation product, sealing all channels of the fermentation tank, and gradually exhausting oxygen from the fermentation product to enter the anaerobic fermentation process;
(8) when the surface bacterial plaque of the liquid fermentation product in the step (7) is in a folded state, the supernatant liquid in the middle layer is golden yellow, clear and transparent, and the solid fermentation product in the bottom layer and the liquid in the middle layer are distinct and are not adhered, namely the solid fermentation-liquid fermentation process is completely finished; the obtained fermentation product can be the prepared biological enzyme preparation.
(9) And (3) gradually heating the solution obtained in the step (8) to 78-88 ℃, cooling the solution for 2 hours, and extracting supernatant of the fermentation product to obtain the complex enzyme preparation.
The step is an inactivation process of the biological enzyme preparation, and the specific operation of the inactivation is as follows:
and (3) gradually heating the mixture obtained in the step (8) to 78-88 ℃ (keeping the temperature at 55 ℃ for one hour after heating, then slowly heating the mixture to 78-88 ℃, introducing cold water into an interlayer of the tank body after two hours in total, cooling the interlayer to the normal temperature, and extracting supernatant of the fermented product to obtain the complex enzyme preparation.
In addition, in the embodiment, the content of the total saponins in the biological enzyme preparation prepared in the step (9) is detected, and the detection result shows that the content of the total saponins in the biological enzyme preparation used by the invention is 1260 mg/L.
Example 2: a method for preparing ginsenoside preparation by biological engineering technology is provided, wherein the ginsenoside preparation is rich in saponin.
Which comprises the following steps:
(1) selecting a compound biological enzyme preparation, wherein the biological enzyme preparation is prepared by fermenting yeast and lactobacillus combined bacteria;
(2) adding high fructose corn syrup F55 into the biological enzyme preparation in the step (1) according to the volume ratio of 4%, and uniformly stirring;
(3) heating the mixture obtained in the step (2) to above 160 ℃;
(4) when the volume of the mixture obtained in the step (2) after evaporation reaches 1/3 of the original mixture, the saponin-rich preparation can be prepared.
The total ginsenoside content of the saponin-rich preparation obtained in step (4) of this example was determined to be 7600 mg/L.
The bio-enzyme preparation and the cultivation of the yeast and lactobacillus combination used for preparing the bio-enzyme preparation in this embodiment are described in example 1, and are not described herein again.
In this example, after the fructose-glucose syrup is added to the biological enzyme preparation prepared in the biological enzyme preparation step (8), and then the biological enzyme preparation is heated and inactivated according to the step (9), the total saponin content after the inactivation of the biological enzyme added with the fructose-glucose syrup is detected to be 1580mg/L, which is slightly higher than that of the biological enzyme preparation directly inactivated in the example 1, and it is assumed that the total saponin concentration may be slightly changed due to a small amount of evaporation of water during inactivation.
Comparing with example 1 and example 2, the total saponin content of the saponin-rich preparation prepared in example 2 is more than 2 times of the total saponin content of the saponin-rich preparation prepared in example 1, which fully indicates that new saponin is generated on the basis of the original total saponin by the reaction of the high fructose corn syrup and the biological enzyme preparation.
Example 3: a method for preparing ginsenoside preparation by biological engineering technology is provided, wherein the ginsenoside preparation is rich in saponin.
Which comprises the following steps:
(1) selecting a compound biological enzyme preparation, wherein the biological enzyme preparation is prepared by fermenting yeast and lactobacillus combined bacteria;
(2) adding the high fructose corn syrup F55 into the biological enzyme preparation in the step (1) according to the volume ratio of 15% and uniformly stirring;
(3) heating the mixture obtained in the step (2) to above 160 ℃;
(4) when the volume of the mixture obtained in the step (2) after evaporation reaches 2/5 of the original mixture, the saponin-rich preparation can be prepared.
The total ginsenoside content of the saponin-rich preparation obtained in step (4) of this example was 18000mg/L, and from this data, it can be seen that the increase in the detected amount of total saponin obtained in step (4) is generated by the reaction between syrup and the bio-enzyme preparation rather than enrichment.
Example 4: a method for preparing ginsenoside preparation by biological engineering technology is provided, wherein the ginsenoside preparation is saponin-rich syrup.
Which comprises the following steps:
(1) selecting a compound biological enzyme preparation, wherein the biological enzyme preparation is prepared by fermenting yeast and lactobacillus combined bacteria;
(2) adding high fructose syrup into the biological enzyme preparation in the step (1) according to the volume ratio of 100% and uniformly stirring;
(3) heating the mixture obtained in the step (2) to above 160 ℃;
(4) when the post-evaporation volume in the mix of step (2) reaches 1/2 of the volume of high fructose corn syrup added, a saponin-rich syrup can be produced.
The total ginsenoside content of the saponin-rich preparation obtained in step (4) of this example was determined to be 16000 mg/L.
The invention adopts a method for preparing ginsenoside by blending and heating biological enzyme preparation and syrup, and the reaction principle is presumed as follows:
the method mainly utilizes rich free active groups (amino acid residues, amide, coenzyme, lactic acid, linoleic acid and other active groups) in a biological enzyme preparation to carry out a series of enzymolysis, acidolysis, hydrolysis, heating catalytic dehydration condensation, acyl modification and the like, and then utilizes the characteristics of polyhydroxy aldehyde and polyhydroxy ketone of monosaccharide and the properties of polyhydroxy reaction, oxidative esterification, acetal reaction, aldehyde group, carbonyl reaction and the like to generate the saponin mainly comprising the saponin Rf and Rg1 through multiple reactions such as heating, catalysis and the like.
Example 5: a method for preparing ginsenoside preparation by biological engineering technology is provided, wherein the ginsenoside preparation is rich in saponin.
Which comprises the following steps:
(1) selecting a compound biological enzyme preparation, wherein the biological enzyme preparation is prepared by fermenting yeast and lactobacillus combined bacteria;
(2) adding the disaccharide-rock candy crystals into the biological enzyme preparation in the step (1) according to the weight ratio of 25%, and uniformly stirring;
(3) heating the mixture obtained in the step (2) to more than 100 ℃, and hydrolyzing the rock sugar to convert the disaccharide into monosaccharide;
(4) heating the mixture obtained in the step (3) to above 160 ℃ for continuous heating and condensation;
(5) when the volume after evaporation in the mixture obtained in the step (4) reaches 2/5 of the volume of the original biological enzyme preparation, the saponin-rich preparation can be prepared.
And (4) detecting the saponin-rich preparation obtained in the step (5), wherein the detection result shows that the total saponin content is 66000 mg/L. This example illustrates that polysaccharides, such as disaccharides such as crystal sugar, granulated sugar, and brown sugar, are reacted with biological enzyme preparations to produce saponins, presumably after heatingFirst by hydrolysis to form monosaccharides, e.g. disaccharide C12H22O11Hydrolyzing to monosaccharide C by heating6H12O6Then the monosaccharide takes part in the reaction to generate the saponin.
The present invention and the embodiments thereof have been described above, and the description is not limiting, and the embodiments shown in the above examples are only one of the embodiments of the present invention, and the practical embodiments are not limited thereto. In summary, those skilled in the art should appreciate that they can readily use the disclosed conception and specific embodiments as a basis for designing or modifying other structures for carrying out the same purposes of the present invention without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (9)
1. A method for preparing ginsenoside preparation by adopting bioengineering technology is characterized in that,
which comprises the following steps:
(1) selecting a compound biological enzyme preparation, wherein the biological enzyme preparation is prepared by fermenting yeast and lactobacillus combined bacteria;
(2) directly heating the biological enzyme preparation in the step (1); or adding syrup, stirring, heating, and concentrating to obtain ginsenoside preparation.
2. The method for preparing a ginsenoside preparation according to claim 1, wherein the ginsenoside preparation comprises at least one of a saponin-rich preparation and a saponin-rich syrup;
which comprises the following steps:
(1) selecting a compound biological enzyme preparation, wherein the biological enzyme preparation is prepared by fermenting yeast and lactobacillus combined bacteria;
(2) adding syrup into the biological enzyme preparation in the step (1) according to the volume ratio of 0-100% and uniformly stirring;
(3) heating and concentrating the mixture obtained in the step (2);
(4) when the volume of the mixture obtained in the step (2) after evaporation reaches 1/3-2/5 of the volume of the original mixture, the saponin-rich preparation can be prepared;
(5) and (3) continuing to heat the saponin-rich preparation obtained in the step (4) until the volume of the evaporated saponin-rich preparation reaches the added amount of the syrup obtained in the step (2), and cooling the material to obtain the saponin-rich syrup.
3. The method for preparing ginsenoside preparations by adopting bioengineering technology according to claim 1 or 2, wherein the preparation steps of the compound biological enzyme preparation are as follows:
(1) selecting yeast and fermentation combined bacteria as strains;
(2) activating strains: moistening potato mud snowflake powder with sterile water according to a weight ratio of 1:1, mixing the strain with the moistened potato mud snowflake powder according to a weight ratio of 1:2, and carrying out aerobic activation in a sterile environment;
(3) preparation and inoculation of the culture medium: the culture medium is prepared by using mashed potato snowflake powder, wheat germ and sticky rice as main materials, adding water into the culture medium, uniformly stirring, steaming, and then airing under an aseptic environment to a temperature below 45 ℃ to finish the preparation of the culture medium;
and (3) mixing the prepared culture medium with the activated strain obtained in the step (2) according to the weight ratio of 10: 1, uniformly mixing;
(4) adjusting the humidity of the culture medium inoculated in the step (3) to ensure that the holding balls are soaked by water and the whole culture medium keeps moist and breathable;
(5) carrying out aerobic fermentation on the culture medium in the step (4) for more than 48 hours; when the culture medium has uniform white and milky plaque visible to naked eyes, transferring the culture medium into a fermentation tank; the solid fermentation substance occupies one third of the fermentation tank, and all channels of the fermentation tank are closed, so that the culture medium fermentation substance entering the tank is continuously subjected to aerobic culture in the tank body until the oxygen in the tank body is exhausted, and the fermentation tank automatically enters an anaerobic fermentation state;
(6) when the pressure in the fermentation tank is unchanged, the solid fermentation product enters saturated fermentation, the strong fermentation fragrance overflows after the fermentation tank is opened, the surface of the solid culture medium is covered with the luxuriant mycelia by visual observation, the fermentation product is taken for detection, and the total number of the viable bacteria of the saccharomycetes is 6.4 multiplied by 104-5.9×105cfu/ml；Total number of lactic acid bacteria 4X 108-5.1×109cfu/ml, at which point the solid state fermentation process has reached saturation, may enter the liquid state fermentation stage;
(7) adding sterile water into the solid-state fermentation product obtained in the step (6) according to the mass ratio of 1:3, fully stirring to dilute the metabolic substrate in the solid-state fermentation process, ensuring that at least 1/5 space is reserved in the fermentation tank body, carrying out aerobic activation culture for more than 72 hours until white or milk-white bacterial plaque is densely distributed on the surface of the liquid-state fermentation product, sealing all channels of the fermentation tank, and gradually exhausting oxygen from the fermentation product to enter the anaerobic fermentation process;
(8) when the surface bacterial plaque of the liquid fermentation product in the step (7) is in a folded state, the supernatant liquid in the middle layer is golden yellow, clear and transparent, and the solid fermentation product in the bottom layer and the liquid in the middle layer are distinct and are not adhered, namely the solid fermentation-liquid fermentation process is completely finished; the obtained fermentation product can be the prepared biological enzyme preparation.
4. The method for preparing a ginsenoside preparation by adopting the bioengineering technology according to claim 3, wherein the preparation step of the compound biological enzyme preparation further comprises the following steps:
(9) and (4) gradually heating the mixture obtained in the step (8) to 78-88 ℃, cooling the mixture after 2 hours, and extracting supernatant of a fermentation product to obtain the biological enzyme preparation.
5. The method for preparing ginsenoside preparations by using bioengineering technology according to claim 4, wherein the yeast and lactobacillus combination in step (1) comprises yeast and lactobacillus, the yeast comprises Candida ethanolica B-JJ1, and the lactobacillus comprises at least one of Lactobacillus buchneri B-JR1, Lactobacillus paracasei B-JR2, Lactobacillus zeae B-JR4, Lactobacillus plantarum B-JR5, and lactobacillus B-JR 6; the preservation number of the Candida ethanolica B-JJ1 is CCTCC NO: M2020136; the preservation number of the lactobacillus buchneri B-JR1 is CCTCC NO: M2021132; the preservation number of the lactobacillus paracasei B-JR2 is CCTCC NO: 2021133; the preservation number of the lactobacillus zeae B-JR4 is CCTCC NO: 2021135; the preservation number of the lactobacillus plantarum B-JR5 is CCTCC NO: 2021501; the preservation number of the lactobacillus B-JR6 is CCTCC NO: M20211502.
6. The method for preparing a ginsenoside preparation according to claim 1 or 2, wherein the syrup is prepared from monosaccharide.
7. The method for preparing ginsenoside preparations by using bioengineering technology according to claim 6, wherein the syrup is high fructose corn syrup.
8. The method of claim 1 or 2, wherein the sugar syrup is prepared from at least one of glucose, sucrose, fructose, maltose, maltobiose, and maltotriose.
9. Use of a ginsenoside preparation of any one of claims 1-8 in the preparation of a beverage product with health promoting effect.
Priority Applications (7)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111224254.8A CN113881750A (en) | 2021-10-19 | 2021-10-19 | A method for preparing ginsenoside preparation by biological engineering technology |
| PCT/CN2022/072964 WO2023065558A1 (en) | 2021-10-19 | 2022-01-20 | Method for preparing ginsenoside preparation by using bioengineering technology |
| CN202211150208.2A CN117286209A (en) | 2021-10-19 | 2022-09-21 | Method for preparing ginsenoside preparation by bioengineering technology and application thereof |
| US18/268,927 US20240043901A1 (en) | 2021-10-19 | 2022-09-29 | A method for preparing ginsenoside preparation by biological engineering technology |
| PCT/CN2022/122383 WO2023066002A1 (en) | 2021-10-19 | 2022-09-29 | Method for preparing ginsenoside preparation by adopting bioengineering technology |
| EP22882613.7A EP4421182A1 (en) | 2021-10-19 | 2022-09-29 | Method for preparing ginsenoside preparation by adopting bioengineering technology |
| ZA2022/11343A ZA202211343B (en) | 2021-10-19 | 2022-10-17 | A method for preparing ginsenoside preparation by biological engineering technology |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111224254.8A CN113881750A (en) | 2021-10-19 | 2021-10-19 | A method for preparing ginsenoside preparation by biological engineering technology |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113881750A true CN113881750A (en) | 2022-01-04 |
Family
ID=79003872
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111224254.8A Withdrawn CN113881750A (en) | 2021-10-19 | 2021-10-19 | A method for preparing ginsenoside preparation by biological engineering technology |
| CN202211150208.2A Pending CN117286209A (en) | 2021-10-19 | 2022-09-21 | Method for preparing ginsenoside preparation by bioengineering technology and application thereof |
Family Applications After (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211150208.2A Pending CN117286209A (en) | 2021-10-19 | 2022-09-21 | Method for preparing ginsenoside preparation by bioengineering technology and application thereof |
Country Status (2)
| Country | Link |
|---|---|
| CN (2) | CN113881750A (en) |
| WO (1) | WO2023065558A1 (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023066002A1 (en) * | 2021-10-19 | 2023-04-27 | 耿胜利 | Method for preparing ginsenoside preparation by adopting bioengineering technology |
| WO2023065558A1 (en) * | 2021-10-19 | 2023-04-27 | 耿胜利 | Method for preparing ginsenoside preparation by using bioengineering technology |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101718465B1 (en) * | 2016-01-08 | 2017-03-21 | 강희철 | Novel Lactobacillus Casei GFC 1 and Extracts from Ginseng-fermented Products Using Lactobacillus Casei GFC 1 and Manufacturing Method Thereof |
| CN106498018B (en) * | 2016-11-07 | 2019-06-07 | 江南大学 | A kind of method that compound bacteria prepares the rare anticancer saponin(e Compound K of ginseng |
| KR101808749B1 (en) * | 2017-10-20 | 2017-12-14 | 주식회사 모든바이오 | Production method of ginsenoside 20(S)-Rg3 and 20(S)-Rh2 using ginsenoside glycosidase |
| CN110959853A (en) * | 2018-09-29 | 2020-04-07 | 安琪酵母股份有限公司 | Rare ginsenoside composition and preparation method and application thereof |
| CN113439838B (en) * | 2020-03-26 | 2022-09-06 | 安琪纽特股份有限公司 | Fermentation method, fermentation product and kit containing fermentation product |
| CN113881750A (en) * | 2021-10-19 | 2022-01-04 | 耿胜利 | A method for preparing ginsenoside preparation by biological engineering technology |
-
2021
- 2021-10-19 CN CN202111224254.8A patent/CN113881750A/en not_active Withdrawn
-
2022
- 2022-01-20 WO PCT/CN2022/072964 patent/WO2023065558A1/en active Application Filing
- 2022-09-21 CN CN202211150208.2A patent/CN117286209A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023066002A1 (en) * | 2021-10-19 | 2023-04-27 | 耿胜利 | Method for preparing ginsenoside preparation by adopting bioengineering technology |
| WO2023065558A1 (en) * | 2021-10-19 | 2023-04-27 | 耿胜利 | Method for preparing ginsenoside preparation by using bioengineering technology |
Also Published As
| Publication number | Publication date |
|---|---|
| CN117286209A (en) | 2023-12-26 |
| WO2023065558A1 (en) | 2023-04-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP3444353B1 (en) | Microbial fermentation of botanicals | |
| CN105054040B (en) | A kind of composition of probiotics fermention ginseng and its preparation method and application | |
| CN109717340B (en) | Fermentation preparation method of two-step cordyceps militaris enzyme combined with composite enzymolysis | |
| KR101409761B1 (en) | A method of preparation for fermented red ginseng using conversion by enzyme mixture and fermentation by lactic acid bacterium and the products containing fermented red ginseng manufactured thereof as effective factor | |
| WO2015143783A1 (en) | Natural conditioning and nourishing ferment and manufacturing method therefor | |
| CN113881750A (en) | A method for preparing ginsenoside preparation by biological engineering technology | |
| JP6722799B2 (en) | Method for producing enzyme solution of low-sugar vegetables and/or fruits | |
| CN104013657A (en) | Post-fermentation extracting method of extracting saponin from American ginseng medicinal material | |
| KR100789261B1 (en) | A fermented ginseng containing deglycosylated ginsenosides and the preparation method thereof | |
| KR101372400B1 (en) | A manufacturing method of red ginseng staf4h with enhanced trace ginsenoside | |
| WO2023066002A1 (en) | Method for preparing ginsenoside preparation by adopting bioengineering technology | |
| CN112868963A (en) | Preparation method and application of ginseng enzyme | |
| CN103614429B (en) | Method for transforming gingko pollen flavonoid glycoside employing microorganisms | |
| CN117281865A (en) | Raw and cooked rhizoma polygonati compound fermentation liquor product and preparation method thereof | |
| Malbaša et al. | Sucrose and inulin balance during tea fungus fermentation | |
| CN111227232A (en) | Preparation method of medlar extract enzyme liquid and product thereof | |
| CN114181842B (en) | Saccharomycopsis fibuligera cx-3 strain for converting anthocyanin to produce protocatechuic acid and application thereof | |
| KR102466825B1 (en) | Lactobacillus acidophilus BLA-9 with biological conversion activity of saponin, a method of preparation for fermented red ginseng extracts having increased content of ginsenoside compound K using the same, fermented red ginseng extracts using thereby and health-functional food composition containing the same | |
| CN115820783A (en) | Preparation method for obtaining multiple triterpenoids by fermenting rosa roxburghii tratt | |
| CN109566995A (en) | A kind of Semen Coicis canned sweet fermented glatinous rice and preparation method thereof | |
| CN111763705B (en) | Preparation method and application of ginsenoside composition | |
| CN113881531A (en) | Functional dendrobium stem fermented rice wine and preparation method thereof | |
| LU506192B1 (en) | A method for preparing ginsenoside preparation by biological engineering technology | |
| Ugale | Preparation of kombucha tea review on: Different kinds of microorganism’s used for development of the kombucha | |
| CN111961093B (en) | Method for extracting oligosaccharide from litchi wine distillation liquid and application of oligosaccharide |
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 | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20220104 |
|
| WW01 | Invention patent application withdrawn after publication |