CN105348337A - Stevioside derivative prepared by stervioside biotransformation, preparation method and application thereof - Google Patents

Stevioside derivative prepared by stervioside biotransformation, preparation method and application thereof Download PDF

Info

Publication number
CN105348337A
CN105348337A CN201510807932.1A CN201510807932A CN105348337A CN 105348337 A CN105348337 A CN 105348337A CN 201510807932 A CN201510807932 A CN 201510807932A CN 105348337 A CN105348337 A CN 105348337A
Authority
CN
China
Prior art keywords
stevioside
bio
preparation
udpg
transformation according
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.)
Granted
Application number
CN201510807932.1A
Other languages
Chinese (zh)
Other versions
CN105348337B (en
Inventor
朱惠霖
丁雪峰
张永正
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
NANJING NUOYUN BIOTECHNOLOGY Co Ltd
Original Assignee
NANJING NUOYUN BIOTECHNOLOGY Co Ltd
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by NANJING NUOYUN BIOTECHNOLOGY Co Ltd filed Critical NANJING NUOYUN BIOTECHNOLOGY Co Ltd
Priority to CN201510807932.1A priority Critical patent/CN105348337B/en
Publication of CN105348337A publication Critical patent/CN105348337A/en
Application granted granted Critical
Publication of CN105348337B publication Critical patent/CN105348337B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
    • C07H15/20—Carbocyclic rings
    • C07H15/24—Condensed ring systems having three or more rings
    • C07H15/256—Polyterpene radicals
    • 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
    • A23L2/60—Sweeteners
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H1/00—Processes for the preparation of sugar derivatives
    • 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
    • C12P19/18—Preparation of compounds containing saccharide radicals produced by the action of a glycosyl transferase, e.g. alpha-, beta- or gamma-cyclodextrins
    • 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
    • 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
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/582—Recycling of unreacted starting or intermediate materials

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Genetics & Genomics (AREA)
  • Biotechnology (AREA)
  • Seasonings (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Polymers & Plastics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Food Science & Technology (AREA)
  • Nutrition Science (AREA)

Abstract

The invention relates to a stevioside derivative prepared by stervioside biotransformation and a preparation method and an application thereof, and belongs to the field of food chemistry. According to the method, stervioside, which is used as a substrate, undergoes enzyme catalysis by the utilization of UDP-glucosyltransferase in the presence of a glucosyl donor under conditions of proper temperature, pH, salinity, substrate concentration and the like, so as to obtain a stevioside derivative NY101 with a brand-new structure. The method for preparing the stevioside derivative NY101 by stervioside biotransformation is simple to operate, is low-cost, and has high transformation efficiency. Transformation efficiency can be greater than 90%. Purification is easy, purity is high, and purity can be greater than 95%. The stevioside derivative NY101 can be used in the food and beverage industry and has huge potential application value.

Description

A kind of stevioside bio-transformation prepare stevioside derivative, preparation method and application thereof
Technical field
The present invention relates to stevioside derivative prepared by a kind of stevioside bio-transformation, preparation method and application thereof, belong to food chemistry technical field.
Background technology
Stevioside is a kind of new type natural sweeting agent extracted from the leaf, stem of catananche's sweet Stevia, be a kind of natural, green, health care functional food, it has fragrance, refrigerant taste, also there is the feature that sugariness is high, heat energy is low, its sugariness is 200-300 times of sucrose, and calorific value is only sucrose volume 1/300, it is the ideal sweeting agent of a kind of alternative sucrose.Along with the mankind are to healthy, green attention, stevioside is called it is future world sweeting agent most with prospects by Food science man.
Stevioside is thought the nutritious supplementary that the mankind are good and health care medicine by international medical community.Prove through a large amount of scientific experiment, stevioside is conducive to regulating blood sugar and blood pressure, often edible regulating blood sugar level, and preventing hypertension, improves hypostension; Stevioside also has the effect suppressing some bacterial growth and breeding and stop it to infect, can treat and catch a cold and the illness such as influenza and preventing dental caries; Stevioside can reduce people to sugar and fatty demand, regulates its diet, reduces people to the demand of tobacco and wine, prevents the generation of the diseases such as diabetes, obesity, heart trouble; Meanwhile, stevioside also has skin repair and improving effect, and it can treat dermatosis, smoothes away wrinkles, and removes scar etc.
Stevioside can be widely used in the industries such as food, beverage, food flavouring, medicine, daily-use chemical industry, wine brewing, makeup, comparatively uses sucrose escapable cost about 60%, and is of high nutritive value.The stability of stevioside, pathways metabolism and security thereof are furtherd investigate, and its Ministry of Health of China, Ministry of Light Industry's approval uses, and also by rank that the accreditation of U.S. food Drug Administration is " GRAS (being commonly considered as safe) ".Stevioside is as the natural substitute of sucrose, and it has a extensive future.
Though stevioside has numerous advantage, but its serious bitter aftertaste, hinder its widespread use.At present, the improvement of stevioside mouthfeel realizes by enzyme process bio-transformation.Have been reported, can utilize Maltose 4-glucosyltransferase modification stevioside, to improve its mouthfeel and taste matter, but the position of this enzyme transfer glycosyl is not single-minded, and product is impure, and its sugariness also seriously reduces.
The two glycosides of stevia rebaudianum is converted into the reaction etc. of rebaudioside A and D to the conversion of stevioside and RB and stevioside.These researchs are for numerous components of stevioside, still very little, and its modified mouthfeel and taste matter are also a huge research.Seek efficient metabolizing enzymes, conversion process and Novel sweet synanthrin derivative are still the research tendency in stevioside future.
Summary of the invention
The technical problem that the present invention solves is: proposing a kind of is raw material with stevioside, stevioside derivative is produced by enzymatic conversion method, thus improve its mouthfeel, the stevioside bio-transformation of cost that can be lower and shorter production cycle output high-quality prepare stevioside derivative, preparation method and application thereof.
In order to solve the problems of the technologies described above, the technical scheme that the present invention proposes is: stevioside derivative prepared by a kind of stevioside bio-transformation, and structural formula is as follows:
Another technical scheme that the present invention proposes is: the preparation method of stevioside derivative prepared by a kind of stevioside bio-transformation: take stevioside as substrate, make described substrate under glucosyl group donor exists, under the katalysis of UDPG based transferase (uridine diphosphoglucose based transferase), reaction generates stevioside derivative.
Preferably, the UDPG reprocessing cycle system that described glucosyl group donor is UDPG or is made up of sucrose, sucrose synthase and UDP.
Preferably, described UDPG based transferase M301, sucrose synthase M302, provided by fermentative production portion of Nanjing Nuoyun Biological Technology Co., Ltd..
Preferably, the consumption of UDPG is 0.0135 ~ 5.41g/L; The consumption of sucrose is 34.23 ~ 68.46g/L; The consumption of sucrose synthase counts 18.92 ~ 94.59g/L by wet thallus,
Preferably, express UDPG based transferase in microorganism host, described microorganism host is selected from intestinal bacteria, bacillus, yeast, aspergillus tubigensis or pichia spp.
Preferably, described reaction is at MgCl 2concentration is 0 ~ 0.286g/L, and temperature is 20 ~ 37 DEG C, carries out in the aqueous phase system of pH5.0 ~ 9.0, and the reaction times is 0.5 ~ 72h.
Preferably, described reaction is carried out in the Tris-HCl damping fluid of pH8.0.
Preferably, the addition of the glycosyltransferase of described reaction is 18.92 ~ 227.03g/L, and substrate initial concentration is 0.676 ~ 54.05g/L, and the addition of coenzyme is 0.0135 ~ 5.41g/L.
Another technical scheme that the present invention proposes is: the application of stevioside derivative prepared by a kind of stevioside bio-transformation: can be applicable to food, beverage, tobacco product, seasonings, daily chemical product, drug component, nutraceutical product, dental health product or makeup.
Beneficial effect of the present invention:
(1) the UDPG reprocessing cycle system that the glucosyl group donor described in can be UDPG or be made up of sucrose, sucrose synthase and UDP.Wherein, the UDPG reprocessing cycle system be preferably made up of sucrose, sucrose synthase and UDP, UDPG is expensive, adopts UDPG reprocessing cycle system significantly can reduce production cost.
(2) join in reaction solution by reaction whole raw materials used, after mixing, under being placed in the parameter of setting, shaking table concussion reaction, at the end of reaction, its product conversion efficiency can reach more than 90%.Stevioside derivative NY101 product is met the requirements of by obtaining the process of reaction solution separating-purifying.A concrete separating and purifying method is first through resin isolation process, then through ethanol periodic crystallisation purifying, according to this method of purification, can obtain the stevioside derivative NY101 product of high purity 95%.
(3) sucrose is as one of raw material, provides glycosyl, because its physicochemical property and stevia rebaudianum carbohydrates and their derivative differ greatly, therefore the mode such as resin isolation and crystallization can be adopted to remove unreacted sucrose and byproduct of reaction fructose after reaction.
(4) the present invention is with stevioside, sucrose for raw material, utilize bioconversion method to react under described parameter and obtain stevioside derivative NY101 product, production technique green safety, and production cost is low, the cycle is short, improves the competitive power of product greatly.Because the present invention is easy and simple to handle, transformation efficiency is high, and purify easily, products obtained therefrom purity is high, can be used for bag and bottle industry, has important using value.
Accompanying drawing explanation
Be described further of the present invention below in conjunction with accompanying drawing.
The HPLC spectrogram of Fig. 1 NY101 and MS spectrogram thereof
The nucleus magnetic hydrogen spectrum figure of Fig. 2 NY101
The nuclear-magnetism carbon spectrogram of Fig. 3 NY101
The two-dimentional COSY spectrogram of Fig. 4 NY101
The two-dimentional HMQC spectrogram of Fig. 5 NY101
The two-dimentional HMBC spectrogram of Fig. 6 NY101
Embodiment
The present invention mainly provides two routes preparing stevioside derivative NY101:
Route one:
Route two:
Embodiment 1:
The structure of recombination bacillus coli and abduction delivering
Molecular biology and genetic engineering technique etc. is utilized to obtain the expression of recombinant e. coli bacterial strain containing goal gene, then by recombination bacillus coli fermentation culture, the reconstitution cell of abduction delivering preparation containing target protein.Its concrete steps are as follows:
1) primer segments needed for synthesis, the UDPG based transferase M301 coding DNA fragment needed for being obtained by pcr amplification, and by homologous recombination technique, be incorporated in the expression cassette of pNYK expression vector.
2) by recombinant plasmid transformed in intestinal bacteria, obtain containing the engineering bacteria J301 of goal gene.
3) 1ml engineering bacteria J301 is placed in TB substratum, 250rpm, 37 DEG C of shaking culture are to OD600=1.0, and adding final concentration is that 0.1mMIPTG is in 25 DEG C of shaking culture 16h.With 10000g, 5min centrifugal collecting cell after induction terminates, be placed in-80 DEG C and store for future use.Obtain the thalline containing M301 albumen.
Same, prepare the thalline containing sucrose synthase M302 albumen.
Embodiment 2:
With stevioside (stevioside) for substrate enzyme process directly prepares NY101 (route one)
Thalline M301 in Example 1 precipitates, and wet thallus is heavily 35mg, uses sterilized water re-suspended cell, and in ice bath, have children outside the state plan ripple smudge cells, is the crude enzyme liquid that reaction is used.Precision takes sample, is mixed with 1.85mL system, and wherein the final concentration 2.70g/L of stevioside (stevioside), UDPG are 2.70g/L, and add the MgCl of 0.286g/L 2; Then add crude enzyme liquid, and to add Tris-HCl damping fluid (0.1M, pH8.0) to system be 1.85ml, initial action.37 DEG C of constant-temperature tables to vibrate 24h with 150rpm, and 100 DEG C are boiled termination reaction.The centrifugal 10min of 13000g, gets supernatant as sample.Use the content of NY101 in LC-MS method detection reaction system.Experimental result shows, the transformation efficiency of NY101 is about 73%.
Embodiment 3:
The determination of best metal ionic concn
Thalline M301 in Example 1 precipitates 35mg, and according to the method in embodiment 2, be transferred in 5ml centrifuge tube, add the stevioside that final concentration is 2.70g/L, UDPG is 2.70g/L and MgCl 2, and add 0.1MTris-HCl damping fluid (pH8.0).As stated above, get Duplicate Samples, add MgCl 2final concentration be respectively 0,0.286g/L, after reaction 24h, sample thief is centrifugal after boiling and stopping, and Supernatant samples is carried out HPLC analysis.Wherein MgCl 2during for 0.1g/L, the output of NY101 is the highest.
Embodiment 4:
The determination of optimal reaction temperature
M301 bacterial sediment 35mg in Example 1, according to the method in embodiment 2, is transferred in 5ml centrifuge tube, and add the Tris-HCl damping fluid (pH8.0) that final concentration is 0.1M, substrate stevioside is 2.70g/L, and UDPG is 2.70g/L.Get Duplicate Samples as stated above, respectively at 20,25,30,37 DEG C, 150rpm carries out catalyzed reaction, and after reaction 24h, sample thief is centrifugal after boiling and stopping, and Supernatant samples is carried out HPLC analysis.When wherein temperature is 30 DEG C, the output of NY101 is the highest.
Embodiment 5:
The determination of optimum response pH
M301 bacterial sediment 35mg in Example 1, according to the method in embodiment 2, is transferred in 5ml centrifuge tube, and add the Tris-HCl damping fluid that final concentration is 0.1M, substrate stevioside is 2.70g/L, and UDPG is 2.70g/L.Get Duplicate Samples as stated above, wherein pH is adjusted to 5.0,6.0,7.0,7.5,8.0,9.0 respectively.In 30 DEG C, 150rpm carries out catalyzed reaction, and after reaction 24h, sample thief is centrifugal, and Supernatant samples is carried out HPLC analysis.When wherein pH is 8.0, the output of NY101 is the highest.
Embodiment 6:
The determination of optimized buffer liquid
Thalline M301 in Example 1 precipitates 35mg, and according to the method in embodiment 2, be transferred in 5ml centrifuge tube, add the substrate stevioside that final concentration is 2.70g/L, UDPG is 2.70g/L.As stated above, get Duplicate Samples, and add the Tris-HCl damping fluid of pH8.0 respectively, phosphate buffered saline buffer, citrate buffer.After reaction 24h, sample thief is centrifugal after boiling and stopping, and Supernatant samples is carried out HPLC analysis.When wherein reacting in Tris-HCl damping fluid, the output of NY101 is the highest.
Embodiment 7:
With stevioside (stevioside) for substrate utilization reprocessing cycle system enzyme legal system is for NY101 (route two)
In the present embodiment, use the UDPG reprocessing cycle system of sucrose, sucrose synthase M302 and UDPG composition as glucosyl group donor.
The each 35mg of M301 and M302 bacterial sediment in Example 1, according to the method in embodiment 2, be transferred in 5ml centrifuge tube, add the Tris-HCl damping fluid (pH8.0) that final concentration is 0.1M, substrate stevioside is 2.70g/L, UDPG is 2.70g/L, and sucrose is 34.23g/L.After adding crude enzyme liquid, initial action.37 DEG C of constant-temperature tables to vibrate 24h with 150rpm, and 100 DEG C are boiled termination reaction.The centrifugal 10min of 13000g, gets supernatant as sample.The content of NY101 in HPLC method detection reaction system.Experimental result shows, the output of NY101 is the highest.
Embodiment 8:
The selection of M301 enzyme addition
Thalline M301 in Example 1 precipitates and M302 bacterial sediment 35mg, according to the method in embodiment 7, be transferred in 5ml centrifuge tube, add the Tris-HCl damping fluid (pH8.0) that final concentration is 0.1M, substrate stevioside is 2.70g/L, UDPG is 2.70g/L, and sucrose is 34.23g/L.As stated above, get Duplicate Samples, and add respectively final concentration be 18.92,37.84,113.51,170.27, the crude enzyme liquid of 227.03g/LM301.After reaction 24h, sample thief is centrifugal after boiling and stopping, and Supernatant samples is carried out HPLC analysis.When wherein adding the reaction of 227.03g/LM301 crude enzyme liquid, the output of NY101 is the highest.
Embodiment 9:
Best substrate addition
The each 35mg of M301 and M302 bacterial sediment in Example 1, according to the method in embodiment 7, is transferred in 5ml centrifuge tube, add the Tris-HCl damping fluid (pH8.0) that final concentration is 0.1M, sucrose is 34.23g/L, and UDPG is 0.676g/L, and substrate stevioside.As stated above, get Duplicate Samples, and add respectively final concentration be 0.676,0.901,1.35,2.70,5.41,13.51,27.03, the stevioside (stevioside) of 54.05g/L.After reaction 24h, sample thief is centrifugal after boiling and stopping, and Supernatant samples is carried out HPLC analysis.When wherein adding 2.70g/L substrate, the output of NY101 is the highest.
Embodiment 10:
Best coenzyme addition
The each 35mg of M301 and M302 bacterial sediment in Example 1, according to the method in embodiment 7, is transferred in 5ml centrifuge tube, add the Tris-HCl damping fluid (pH8.0) that final concentration is 0.1M, substrate stevioside is 2.70g/L, and sucrose is 34.23g/L, and UDPG.As stated above, get Duplicate Samples, and add respectively final concentration be 5.41,2.70,1.35,0.676,0.270,0.135,0.0541,0.0270, the coenzyme of 0.0135g/L.After reaction 24h, sample thief is centrifugal after boiling and stopping, and Supernatant samples is carried out HPLC analysis.When wherein adding 0.676g/L coenzyme, the output of NY101 is the highest.
Embodiment 11:
NY101 structural analysis
Get appropriate NY101, be dissolved in deuterated pyridine, parameters, carry out hydrogen, carbon and ID NMR speetna analysis, resolve NY101 structure according to mutual relationship hydrocarbon in nuclear magnetic spectrogram.
Above-described embodiment, only for technical conceive of the present invention and feature thereof are described, are intended to allow those skilled in the art easy understand content of the present invention also implement according to this, can not limit the scope of the invention with this.In every case the equivalence done according to spirit of the present invention changes or modifies, and all should be encompassed within protection scope of the present invention.

Claims (10)

1. the stevioside derivative prepared of stevioside bio-transformation, is characterized in that: structural formula is as follows:
2. the preparation method of stevioside derivative for preparing of stevioside bio-transformation according to claim 1, it is characterized in that: take stevioside as substrate, make described substrate under glucosyl group donor exists, under the katalysis of UDPG based transferase, reaction generates stevioside derivative.
3. the preparation method of stevioside derivative for preparing of stevioside bio-transformation according to claim 1, is characterized in that: the UDPG reprocessing cycle system that described glucosyl group donor is UDPG or is made up of sucrose, sucrose synthase and UDP.
4. the preparation method of stevioside derivative prepared by the stevioside bio-transformation according to Claims 2 or 3, is characterized in that: described UDPG based transferase, and sucrose synthase is all provided by fermentative production portion of Nanjing Nuoyun Biological Technology Co., Ltd..
5. the preparation method of stevioside derivative prepared by the stevioside bio-transformation according to Claims 2 or 3, is characterized in that: the consumption of described UDPG is 0.0135 ~ 5.41g/L; The consumption of sucrose is 34.23 ~ 68.46g/L; The consumption of sucrose synthase counts 18.92 ~ 94.59g/L by wet thallus.
6. the preparation method of stevioside derivative prepared by the stevioside bio-transformation according to Claims 2 or 3, it is characterized in that: in microorganism host, express UDPG based transferase, described microorganism host is selected from intestinal bacteria, bacillus, yeast, aspergillus tubigensis or pichia spp.
7. the preparation method of stevioside derivative prepared by the stevioside bio-transformation according to Claims 2 or 3, is characterized in that: described reaction is at MgCl 2concentration is 0 ~ 0.286g/L, and temperature is 20 ~ 37 DEG C, carries out in the aqueous phase system of pH5.0 ~ 9.0, and the reaction times is 0.5 ~ 72h.
8. the preparation method of stevioside derivative prepared by the stevioside bio-transformation according to Claims 2 or 3, is characterized in that: described reaction is carried out in the Tris-HCl damping fluid of pH8.0.
9. the preparation method of stevioside derivative prepared by the stevioside bio-transformation according to Claims 2 or 3, it is characterized in that: the addition of the glycosyltransferase of described reaction is 18.92 ~ 227.03g/L, substrate initial concentration is 0.676 ~ 54.05g/L, and the addition of coenzyme is 0.0135 ~ 5.41g/L.
10. the application of stevioside derivative prepared of stevioside bio-transformation according to claim 1, is characterized in that: can be applicable to food, beverage, tobacco product, seasonings, daily chemical product, drug component, nutraceutical product, dental health product or makeup.
CN201510807932.1A 2015-11-19 2015-11-19 Stevia rebaudianum sugar derivatives prepared by a kind of stevioside bioconversion, preparation method and applications Active CN105348337B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201510807932.1A CN105348337B (en) 2015-11-19 2015-11-19 Stevia rebaudianum sugar derivatives prepared by a kind of stevioside bioconversion, preparation method and applications

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201510807932.1A CN105348337B (en) 2015-11-19 2015-11-19 Stevia rebaudianum sugar derivatives prepared by a kind of stevioside bioconversion, preparation method and applications

Publications (2)

Publication Number Publication Date
CN105348337A true CN105348337A (en) 2016-02-24
CN105348337B CN105348337B (en) 2018-07-27

Family

ID=55324418

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201510807932.1A Active CN105348337B (en) 2015-11-19 2015-11-19 Stevia rebaudianum sugar derivatives prepared by a kind of stevioside bioconversion, preparation method and applications

Country Status (1)

Country Link
CN (1) CN105348337B (en)

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105732740A (en) * 2016-03-28 2016-07-06 南京诺云生物科技有限公司 Preparation method of novel sweetening agent
CN105777824A (en) * 2016-03-28 2016-07-20 南京诺云生物科技有限公司 Stevioside derivative and application of stevioside derivative
CN105803021A (en) * 2016-03-28 2016-07-27 南京诺云生物科技有限公司 Preparing method for sweetening agent
CN105838759A (en) * 2016-03-28 2016-08-10 南京诺云生物科技有限公司 Preparation method of stevia sugar derivative
CN108103127A (en) * 2017-12-13 2018-06-01 桂林莱茵生物科技股份有限公司 A kind of method for industrializing quick production and preparing glucosyl group steviol glycoside mixture
CN110669747A (en) * 2019-11-12 2020-01-10 广西师范大学 Enzyme for preparing enzyme-modified stevioside and application thereof
CN111073923A (en) * 2018-10-22 2020-04-28 山东三元生物科技股份有限公司 Enzymatic preparation method of rebaudioside-M
CN111285909A (en) * 2018-12-06 2020-06-16 中国医学科学院药物研究所 Optically active bicyclol glucoside, preparation method thereof and application of optically active bicyclol glucoside in preventing and treating liver diseases
CN116462777A (en) * 2023-04-14 2023-07-21 桂林莱茵合成生物技术有限公司 Novel glucosyl stevioside RMM and application and synthesis method thereof

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1852661A (en) * 2003-08-14 2006-10-25 嘉吉有限公司 Chewing gum compositions comprising monatin and methods for making same
WO2015065650A2 (en) * 2013-11-01 2015-05-07 Conagen Inc. Recombinant production of steviol glycosides

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1852661A (en) * 2003-08-14 2006-10-25 嘉吉有限公司 Chewing gum compositions comprising monatin and methods for making same
WO2015065650A2 (en) * 2013-11-01 2015-05-07 Conagen Inc. Recombinant production of steviol glycosides

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105732740A (en) * 2016-03-28 2016-07-06 南京诺云生物科技有限公司 Preparation method of novel sweetening agent
CN105777824A (en) * 2016-03-28 2016-07-20 南京诺云生物科技有限公司 Stevioside derivative and application of stevioside derivative
CN105803021A (en) * 2016-03-28 2016-07-27 南京诺云生物科技有限公司 Preparing method for sweetening agent
CN105838759A (en) * 2016-03-28 2016-08-10 南京诺云生物科技有限公司 Preparation method of stevia sugar derivative
CN108103127A (en) * 2017-12-13 2018-06-01 桂林莱茵生物科技股份有限公司 A kind of method for industrializing quick production and preparing glucosyl group steviol glycoside mixture
CN111073923A (en) * 2018-10-22 2020-04-28 山东三元生物科技股份有限公司 Enzymatic preparation method of rebaudioside-M
CN111285909A (en) * 2018-12-06 2020-06-16 中国医学科学院药物研究所 Optically active bicyclol glucoside, preparation method thereof and application of optically active bicyclol glucoside in preventing and treating liver diseases
CN110669747A (en) * 2019-11-12 2020-01-10 广西师范大学 Enzyme for preparing enzyme-modified stevioside and application thereof
CN116462777A (en) * 2023-04-14 2023-07-21 桂林莱茵合成生物技术有限公司 Novel glucosyl stevioside RMM and application and synthesis method thereof
CN116462777B (en) * 2023-04-14 2023-11-17 桂林莱茵合成生物技术有限公司 Novel glucosyl stevioside RMM and application and synthesis method thereof

Also Published As

Publication number Publication date
CN105348337B (en) 2018-07-27

Similar Documents

Publication Publication Date Title
CN105348337A (en) Stevioside derivative prepared by stervioside biotransformation, preparation method and application thereof
CN103397064B (en) Method for preparing rebaudioside M through enzyme method
CN104726523B (en) A kind of method that enzyme process prepares Rebaudiodside A M
CN103757074B (en) A kind of enzyme process prepares the method for rebaudioside M
CN102492757B (en) Method for improving taste quality of stevioside by using beta-cyclodextrin glucosyltransferase
CN109750071A (en) A kind of method of biocatalysis synthesis Rebaudiodside A M
CN106471128A (en) A kind of method that enzyme process prepares rebaudioside M
WO2017000366A1 (en) Method for preparing rebaudioside m by using saccharomyces cerevisiae enzymatic process
CN107058200B (en) Method for preparing L-ascorbic acid-2-glucoside
WO2018072213A1 (en) Method for preparing rebaudioside n using enzymatic method
CN105838759A (en) Preparation method of stevia sugar derivative
CN105255805B (en) A kind of Bacillus subtilis genes engineered strain and its construction method and the application in production lactulose
CN108013306A (en) The preparation method of juice drinks rich in psicose
CN105380224A (en) Novel stevioside derivative, preparation method and application thereof
CN109402154B (en) Method for improving isoorientin yield of recombinant bacteria by utilizing cooperative regulation and control strategy
CN103205391A (en) Gene engineering strain and application thereof
CN105219663A (en) The special strain therefore of trehalose synthesis and the method for the synthesis of trehalose thereof
CN105732740A (en) Preparation method of novel sweetening agent
CN105777824A (en) Stevioside derivative and application of stevioside derivative
CN104480056B (en) A kind of genetic engineering bacterium of high-yield extracellular polysaccharide and its preparation method and application
CN105820199A (en) Preparation method for novel stevioside derivative
CN105777823A (en) Sweetener and application of sweetener
CN105803021A (en) Preparing method for sweetening agent
CN101603064A (en) A kind of method for preparing D-tagatose and L-tagatose by galactitol
CN105154459A (en) Novel bacillus arabinose isomerase gene cloning and expression method and use

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant