CN111073923A - Enzymatic preparation method of rebaudioside-M - Google Patents
Enzymatic preparation method of rebaudioside-M Download PDFInfo
- Publication number
- CN111073923A CN111073923A CN201811228403.6A CN201811228403A CN111073923A CN 111073923 A CN111073923 A CN 111073923A CN 201811228403 A CN201811228403 A CN 201811228403A CN 111073923 A CN111073923 A CN 111073923A
- Authority
- CN
- China
- Prior art keywords
- rebaudioside
- glycosyltransferase
- stevioside
- cyclodextrin
- udp
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- GSGVXNMGMKBGQU-PHESRWQRSA-N rebaudioside M Chemical compound C[C@@]12CCC[C@](C)([C@H]1CC[C@@]13CC(=C)[C@@](C1)(CC[C@@H]23)O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O[C@@H]2O[C@H](CO)[C@@H](O)[C@H](O)[C@H]2O)[C@H]1O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O)C(=O)O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O[C@@H]2O[C@H](CO)[C@@H](O)[C@H](O)[C@H]2O)[C@H]1O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O GSGVXNMGMKBGQU-PHESRWQRSA-N 0.000 title claims abstract description 19
- 230000002255 enzymatic effect Effects 0.000 title claims description 11
- 238000002360 preparation method Methods 0.000 title claims description 10
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims abstract description 21
- 235000019202 steviosides Nutrition 0.000 claims abstract description 21
- UEDUENGHJMELGK-HYDKPPNVSA-N Stevioside Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@]12C(=C)C[C@@]3(C1)CC[C@@H]1[C@@](C)(CCC[C@]1([C@@H]3CC2)C)C(=O)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O UEDUENGHJMELGK-HYDKPPNVSA-N 0.000 claims abstract description 20
- 229940013618 stevioside Drugs 0.000 claims abstract description 20
- OHHNJQXIOPOJSC-UHFFFAOYSA-N stevioside Natural products CC1(CCCC2(C)C3(C)CCC4(CC3(CCC12C)CC4=C)OC5OC(CO)C(O)C(O)C5OC6OC(CO)C(O)C(O)C6O)C(=O)OC7OC(CO)C(O)C(O)C7O OHHNJQXIOPOJSC-UHFFFAOYSA-N 0.000 claims abstract description 20
- 102000004190 Enzymes Human genes 0.000 claims abstract description 19
- 108090000790 Enzymes Proteins 0.000 claims abstract description 19
- 108090000992 Transferases Proteins 0.000 claims abstract description 13
- 102000004357 Transferases Human genes 0.000 claims abstract description 13
- 238000000034 method Methods 0.000 claims abstract description 12
- 108010025880 Cyclomaltodextrin glucanotransferase Proteins 0.000 claims abstract description 11
- 125000001312 palmitoyl group Chemical group O=C([*])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 claims abstract description 11
- 229920000858 Cyclodextrin Polymers 0.000 claims abstract description 10
- 239000001116 FEMA 4028 Substances 0.000 claims abstract description 10
- WHGYBXFWUBPSRW-FOUAGVGXSA-N beta-cyclodextrin Chemical compound OC[C@H]([C@H]([C@@H]([C@H]1O)O)O[C@H]2O[C@@H]([C@@H](O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O3)[C@H](O)[C@H]2O)CO)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H]3O[C@@H]1CO WHGYBXFWUBPSRW-FOUAGVGXSA-N 0.000 claims abstract description 10
- 235000011175 beta-cyclodextrine Nutrition 0.000 claims abstract description 10
- 229960004853 betadex Drugs 0.000 claims abstract description 10
- 239000002994 raw material Substances 0.000 claims abstract description 8
- 125000003147 glycosyl group Chemical group 0.000 claims abstract description 6
- 238000006243 chemical reaction Methods 0.000 claims description 18
- 239000000758 substrate Substances 0.000 claims description 4
- -1 acetoacetyl ester Chemical class 0.000 claims description 3
- 239000007853 buffer solution Substances 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims description 2
- 230000035484 reaction time Effects 0.000 claims description 2
- 241000544066 Stevia Species 0.000 description 7
- HELXLJCILKEWJH-NCGAPWICSA-N rebaudioside A Chemical compound O([C@H]1[C@H](O)[C@@H](CO)O[C@H]([C@@H]1O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)O[C@]12C(=C)C[C@@]3(C1)CC[C@@H]1[C@@](C)(CCC[C@]1([C@@H]3CC2)C)C(=O)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O HELXLJCILKEWJH-NCGAPWICSA-N 0.000 description 6
- 230000000694 effects Effects 0.000 description 4
- 235000013305 food Nutrition 0.000 description 4
- 229930182470 glycoside Natural products 0.000 description 4
- 244000228451 Stevia rebaudiana Species 0.000 description 3
- 235000006092 Stevia rebaudiana Nutrition 0.000 description 3
- HSCJRCZFDFQWRP-JZMIEXBBSA-N UDP-alpha-D-glucose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1OP(O)(=O)OP(O)(=O)OC[C@@H]1[C@@H](O)[C@@H](O)[C@H](N2C(NC(=O)C=C2)=O)O1 HSCJRCZFDFQWRP-JZMIEXBBSA-N 0.000 description 3
- HSCJRCZFDFQWRP-UHFFFAOYSA-N Uridindiphosphoglukose Natural products OC1C(O)C(O)C(CO)OC1OP(O)(=O)OP(O)(=O)OCC1C(O)C(O)C(N2C(NC(=O)C=C2)=O)O1 HSCJRCZFDFQWRP-UHFFFAOYSA-N 0.000 description 3
- 235000003599 food sweetener Nutrition 0.000 description 3
- 150000002338 glycosides Chemical class 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 239000003765 sweetening agent Substances 0.000 description 3
- 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 2
- 229930006000 Sucrose Natural products 0.000 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 description 2
- 235000013361 beverage Nutrition 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 229960004793 sucrose Drugs 0.000 description 2
- 241000208838 Asteraceae Species 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 240000004670 Glycyrrhiza echinata Species 0.000 description 1
- 235000001453 Glycyrrhiza echinata Nutrition 0.000 description 1
- 235000006200 Glycyrrhiza glabra Nutrition 0.000 description 1
- 235000017382 Glycyrrhiza lepidota Nutrition 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 101710204244 Processive diacylglycerol beta-glucosyltransferase Proteins 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- 235000008452 baby food Nutrition 0.000 description 1
- 230000002210 biocatalytic effect Effects 0.000 description 1
- 235000019658 bitter taste Nutrition 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 238000011033 desalting Methods 0.000 description 1
- XYIBRDXRRQCHLP-UHFFFAOYSA-N ethyl acetoacetate Chemical compound CCOC(=O)CC(C)=O XYIBRDXRRQCHLP-UHFFFAOYSA-N 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 235000008216 herbs Nutrition 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 201000001421 hyperglycemia Diseases 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 229940010454 licorice Drugs 0.000 description 1
- 235000021096 natural sweeteners Nutrition 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 235000019640 taste Nutrition 0.000 description 1
- 239000002351 wastewater Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 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
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/44—Preparation of O-glycosides, e.g. glucosides
- C12P19/56—Preparation of O-glycosides, e.g. glucosides having an oxygen atom of the saccharide radical directly bound to a condensed ring system having three or more carbocyclic rings, e.g. daunomycin, adriamycin
Landscapes
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a method for preparing rebaudioside M by an enzyme method, which is mainly characterized in that stevioside is used as a raw material, ethyl acetate and β -cyclodextrin are used as glycosyl donors, palmitoyl transferase, cyclodextrin glycosyl transferase and UDP-glycosyl transferase are used as main tool enzymes to generate the rebaudioside M.
Description
Technical Field
The invention discloses a method for preparing rebaudioside-M from stevioside serving as a raw material, ethyl acetate and β -cyclodextrin serving as glycosyl donors, and enzyme components consisting of palmitoyl transferase, cyclodextrin glycosyltransferase and UDP-glycosyltransferase serving as main tool enzymes, and belongs to the technical field of biocatalytic conversion.
Background
Steviosides (stevia glycosides) are natural sweeteners extracted from the leaves of stevia Rebaudiana Bertoni of Compositae herbs. Is a mixture of various glycosides, and different glycosides have large differences in taste quality. Stevioside has high sweetness, low calorie, high stability, potential curative effects of resisting hyperglycemia, hypertension, inflammation, tumor, diarrhea and the like, and effects of regulating immunity and the like, and has attracted extensive attention as a sweetener for related research and economy in food and beverage. High purity stevia products have been officially approved as food additives in the United states and the European Union.
In recent years, over ten glycosides have been isolated from stevia rebaudiana. Rebaudioside M (Rebaudianide M, RebM) is the most potential stevioside, has high sweetness which is about 300 times that of cane sugar and 350 times that of other stevioside, has pure sweetness and better mouthfeel which is close to cane sugar, has no sweetness and bitterness and licorice peculiar smell, and has good stability, thereby being an ideal natural high-power sweetener product. In 2013, the FDA in the united states approved the use of a stevia product extracted from stevia leaves containing Reb M at a content of 50%, and recognized its general safety (GRAS). In 2014, the us FDA was approved for use in beverages and foods other than infant food by GRAS approval containing 95% content Reb M extracted from stevia leaves. In 2015, Reb M was approved by the australian new zealand food standards agency as a sweetener for food.
The content of Reb M in stevia leaves is very low (less than 1%) and is only detected in stevia Morita plants. The Reb M produced by the extraction method needs a large amount of stevia rebaudiana raw materials, in addition, the process for enriching the Reb M is complicated, column passing, desalting, decoloring and recrystallizing are needed for many times after extraction, a large amount of wastewater is generated in the production process, the production cost is high, and the method is not suitable for industrial mass production.
The prior method for synthesizing Reb M by a biological enzyme method needs to add expensive UDP-glucose as a substrate, and takes stevioside, Reb A, Reb D and the like as the substrates to generate the Reb M by catalysis under the action of UDP-glucosyltransferase. But due to the extremely high price of UDP-glucose, the feasibility of industrial preparation of Reb M is almost completely limited, the economy is poor, and the market competitiveness is lacked.
Disclosure of Invention
The problem to be solved by the invention is to provide a method for preparing rebaudioside M by an enzyme method, the method takes stevioside as a raw material, ethyl acetate and β -cyclodextrin as glycosyl donors, and enzyme components consisting of palmitoyl transferase, cyclodextrin glycosyltransferase and UDP-glycosyltransferase as main tool enzymes to generate rebaudioside M.
In order to achieve the above objects and the objects of the present invention, the present invention provides the following technical solutions:
the invention is characterized in that stevioside is used as a raw material, ethyl acetate and β -cyclodextrin are used as glycosyl donors, enzyme components consisting of palmitoyl transferase, cyclodextrin glycosyltransferase and UDP-glycosyltransferase are used as main tool enzymes, and rebaudioside M is generated.
The enzymatic preparation method of rebaudioside M is characterized in that the initial reaction concentration of the substrate raw material stevioside is 2-30 g/L, the mass ratio of the ethyl acetoacetate to the stevioside is 5-10: 1, the mass ratio of β -cyclodextrin to the stevioside is 5-10: 1, and the mass ratio of an enzyme component consisting of palmitoyl transferase, cyclodextrin glycosyltransferase and UDP-glycosyltransferase to the stevioside is 0.05: 1.
The enzyme component is characterized in that the enzyme activity ratio of palmitoyl transferase, cyclodextrin glycosyltransferase and UDP-glycosyltransferase is 2:1:4, wherein the enzyme activity of UDP-glycosyltransferase is 100-200U/mL.
The above-mentioned enzymatic preparation method of rebaudioside-M is characterized in that the conversion reaction employs an aqueous phase system, and the reaction system performs biological conversion in a buffer solution with pH 6-8.
The enzymatic preparation method of rebaudioside-M is characterized in that the reaction temperature of the conversion reaction is 20-50 ℃, and the reaction time is 1-24 h.
Through the technical scheme, the use of expensive glycosyl donors can be avoided, the cost of the used glucose glycosyl donors, namely ethyl acetate and β -cyclodextrin is far lower than that of UDP-glucose, and the actual test shows that enzyme components consisting of palmitoyl transferase, cyclodextrin glycosyl transferase and UDP-glycosyl transferase used in the invention can add glucose on β -cyclodextrin to corresponding sites of stevioside under the condition of the existence of ethyl acetate in the proportion, so that the stevioside is converted into Reb M, and the aim of the invention is fulfilled.
Detailed Description
Example 1 an aqueous reaction system was prepared, the initial reaction concentration of stevioside was 20 g/L, the initial reaction concentration of acetoacetyl ester was 100 g/L, the initial reaction concentration of β -cyclodextrin was 100 g/L, the initial reaction concentration of palmitoyl transferase was 57U/mL, the initial reaction concentration of cyclodextrin glycosyltransferase was 28U/mL, and the initial reaction concentration of UDP-glycosyltransferase was 114U/mL, and the reaction system was adjusted to pH 6-8 using a phosphate buffer.
Example 2 the aqueous reaction system of example 1 was placed in a constant temperature shaker at 30 ℃ for 24 h. Thus obtaining the water solution containing rebaudioside M, and determining the concentration of rebaudioside M to be 7.5g/L by using a high performance liquid chromatography method.
Claims (5)
1. An enzymatic preparation method of rebaudioside-M is characterized in that rebaudioside-M is generated by taking stevioside as a raw material, taking ethyl acetate and β -cyclodextrin as glycosyl donors and taking enzyme components consisting of palmitoyl transferase, cyclodextrin glycosyltransferase and UDP-glycosyltransferase as main tool enzymes.
2. The enzymatic preparation method of rebaudioside M according to claim 1, characterized in that the initial reaction concentration of the substrate raw material stevioside is 2-30 g/L, the mass ratio of the acetoacetyl ester to the stevioside is 5-10: 1, the mass ratio of β -cyclodextrin to the stevioside is 5-10: 1, and the mass ratio of the enzyme component composed of palmitoyl transferase, cyclodextrin glycosyltransferase, UDP-glycosyltransferase to the stevioside is 0.05: 1.
3. The enzyme composition according to claim 1, wherein the ratio of the enzymatic activities of palmitoyl transferase, cyclodextrin glycosyltransferase, and UDP-glycosyltransferase is 2:1:4, wherein the enzymatic activity of UDP-glycosyltransferase is 100-200U/mL.
4. The method for the enzymatic preparation of rebaudioside M according to claim 1, wherein said conversion reaction is carried out in an aqueous system and the reaction system is a biological conversion in a buffer solution having a pH of 6-8.
5. The enzymatic preparation method of rebaudioside M according to claim 1, wherein the reaction temperature of the conversion reaction is 20 to 50 ℃ and the reaction time is 1 to 24 hours.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811228403.6A CN111073923A (en) | 2018-10-22 | 2018-10-22 | Enzymatic preparation method of rebaudioside-M |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811228403.6A CN111073923A (en) | 2018-10-22 | 2018-10-22 | Enzymatic preparation method of rebaudioside-M |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111073923A true CN111073923A (en) | 2020-04-28 |
Family
ID=70309689
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811228403.6A Pending CN111073923A (en) | 2018-10-22 | 2018-10-22 | Enzymatic preparation method of rebaudioside-M |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111073923A (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103397064A (en) * | 2013-08-14 | 2013-11-20 | 苏州汉酶生物技术有限公司 | Method for preparing rebaudioside M through enzyme method |
| CN104163839A (en) * | 2014-07-04 | 2014-11-26 | 苏州景泓生物技术有限公司 | Process for preparing rebaudioside M |
| CN104341470A (en) * | 2014-10-17 | 2015-02-11 | 成都华高瑞甜科技有限公司 | Synthesis method of rebaudioside M, intermediate product of rebaudioside M and synthesis method of intermediate product of rebaudioside M |
| CN104726523A (en) * | 2015-03-28 | 2015-06-24 | 南京工业大学 | Method for preparing rebaudioside M by enzyme method |
| CN105348337A (en) * | 2015-11-19 | 2016-02-24 | 南京诺云生物科技有限公司 | Stevioside derivative prepared by stervioside biotransformation, preparation method and application thereof |
| WO2016054534A1 (en) * | 2014-10-03 | 2016-04-07 | Conagen Inc. | Non-caloric sweeteners and methods for synthesizing |
-
2018
- 2018-10-22 CN CN201811228403.6A patent/CN111073923A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103397064A (en) * | 2013-08-14 | 2013-11-20 | 苏州汉酶生物技术有限公司 | Method for preparing rebaudioside M through enzyme method |
| CN104163839A (en) * | 2014-07-04 | 2014-11-26 | 苏州景泓生物技术有限公司 | Process for preparing rebaudioside M |
| WO2016054534A1 (en) * | 2014-10-03 | 2016-04-07 | Conagen Inc. | Non-caloric sweeteners and methods for synthesizing |
| CN104341470A (en) * | 2014-10-17 | 2015-02-11 | 成都华高瑞甜科技有限公司 | Synthesis method of rebaudioside M, intermediate product of rebaudioside M and synthesis method of intermediate product of rebaudioside M |
| CN104726523A (en) * | 2015-03-28 | 2015-06-24 | 南京工业大学 | Method for preparing rebaudioside M by enzyme method |
| CN105348337A (en) * | 2015-11-19 | 2016-02-24 | 南京诺云生物科技有限公司 | Stevioside derivative prepared by stervioside biotransformation, preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| OLSSON, KIM等: "Microbial production of next-generation stevia sweeteners", 《MICROBIAL CELL FACTORIES》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| AU2018413277B2 (en) | High-purity steviol glycosides | |
| US10273519B2 (en) | Diterpene production in Yarrowia | |
| CN107666834B (en) | Steviol glycosides | |
| BR112016002092B1 (en) | STEVIOL GLYCOSIDE RECOVERY | |
| JP2022530706A (en) | Uridine diphosphate-dependent glycosyltransferase | |
| JP7634477B2 (en) | High Purity Steviol Glycosides | |
| JP2024133514A (en) | Highly pure steviol glycosides | |
| CN112513059A (en) | High purity steviol glycosides | |
| EP2957182A1 (en) | Improved natural sweetener compositions | |
| CN111093390A (en) | High purity steviol glycosides | |
| KR20210129679A (en) | High Purity Steviol Glycoside | |
| WO2019178471A1 (en) | High-purity steviol glycosides | |
| CN112512337A (en) | High purity steviol glycosides | |
| KR20230112673A (en) | Glycoside product biosynthesis and recovery | |
| BR112018000298B1 (en) | Method for reducing the kaurenoic acid content of a steviol glycoside composition | |
| CN110656149A (en) | Preparation method of rebaudioside D, and product and application thereof | |
| CN111073923A (en) | Enzymatic preparation method of rebaudioside-M | |
| CN116848239A (en) | Microorganism for diterpene production | |
| US20240360489A1 (en) | High-purity steviol glycosides | |
| WO2024228615A1 (en) | High-purity steviol glycosides | |
| WO2024010442A1 (en) | High-purity steviol glycosides | |
| WO2025048633A2 (en) | High-purity steviol glycosides | |
| TW202446949A (en) | Lactobacillus mali strain and uses thereof | |
| BR112017025614B1 (en) | METHOD FOR PRODUCING STEVIOL GLYCOSIDE AND COMPOSITION |
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 | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20200428 |
|
| WD01 | Invention patent application deemed withdrawn after publication |