CN104593450A - Method for splitting pectin with high esterification degree to prepare monosaccharide and oligogalacturonic acid employing pectinase - Google Patents
Method for splitting pectin with high esterification degree to prepare monosaccharide and oligogalacturonic acid employing pectinase Download PDFInfo
- Publication number
- CN104593450A CN104593450A CN201510037006.0A CN201510037006A CN104593450A CN 104593450 A CN104593450 A CN 104593450A CN 201510037006 A CN201510037006 A CN 201510037006A CN 104593450 A CN104593450 A CN 104593450A
- Authority
- CN
- China
- Prior art keywords
- pectin
- monose
- polygalacturonase
- galacturonic acid
- acid oligosaccharides
- 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
Links
Classifications
-
- 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
- Y02P20/52—Improvements relating to the production of bulk chemicals using catalysts, e.g. selective catalysts
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a method for splitting pectin with high esterification degree to prepare monosaccharide and oligogalacturonic acid employing pectinase. The method comprises the following steps: (1) carrying out fermenting cultivation on a GS115-pel168s-9k strain to prepare a pectin enzyme liquid; (2) dissolving 10g of pectin into a buffer solution with the pH of 9.4, adjusting the pH to be 8.8-9.2, preheating to 50 DEG C in a water bath, adding 110U of pectin enzyme liquid and reacting for two hours, so as to obtain enzymatic hydrolysate; and (3) centrifuging the enzymatic hydrolysate prepared from the step (2), evaporating and drying supernate to obtain the monosaccharide and the oligogalacturonic acid, and calculating the hydrolysis rate which can reach 93%. The method has the advantages of simple process, low cost and high enzymatic hydrolysis efficiency.
Description
Technical field
The invention belongs to function technical field of organic matter preparation, the enzymatic lysis height gamma value pectin particularly utilizing the pichia spp fermentation of producing polygalacturonase to produce produces the method for monose and galacturonic acid oligosaccharides.
Background technology
Pectin (pectin) is extensively distributed among the root of fruit, stem, leaf with the form of protopectin-, pectic acid, there is the features such as natural, green, nutrition, health, be through the food ingredients that is nontoxic, that limit without ADI (acceptable daily intake) that Food and Agricultural Organization of the United Nations and the World Health Organization (FAO/WHO) the joint specialist council admit jointly.At present, pectin is widely used in foodstuffs industry as thickening material, stablizer, gelifying agent and synergistic agent etc.In addition, pectin also can be used for protective foods and medicine.Pectin has reduction blood sugar prevent diabetes, reduces cholesterol, prevents colorectal carcinoma and prostate cancer, prevents obesity and suppress the effect of intestinal pathogenic propagation, therefore may be used for making the protective foods preventing the diseases such as diabetes, obesity, hyperlipidemia.In pharmaceutical industries, pectin also has the effect such as antibacterial, hemostasis, reducing blood-fat and radioprotective, separately or can share preparation ointment with other preparation, the pharmaceutical preparation such as suppository and micro-capsule, be called as " balancing agent of HUMAN HEALTH ".
But pectin limits many-sided application such as its biological function because its molecular weight is too large.In addition, different sources, raw material type, growing environment and the difference in season also to make in its structure different, also determine its difference on functional performance, also limit its application commercially, therefore carry out modification to pectin and become a study hotspot.Can obtain a large amount of galacturonic acid oligosaccharides by depolymerized pectin, degradation pathway theoretical at present has physical degradation methods, chemical degradation method and enzyme liberating method.Although with Physical (as Zhang Lifen, pectin polysaccharide ultrasonic directional degradation pathway and study mechanism thereof, Zhejiang University doctor academic dissertation, 2013), chemical method (acid system depolymerized pectin) also can make its cracking, but there is the problem such as unstable products, contaminate environment in physics and chemistry method.Then there are not the problems referred to above with enzyme liberating, polygalacturonase is can general name (Xue Changhu, the Zhang Yongqin of multiple enzyme of decompose pectin material, Li Zhaojie, Li Zhijun, the progress of pectin and polygalacturonase, food and biotechnology journal, 06 phase in 2005), carry out crudefiber crop with enzyme liberating pectin to come unstuck, the existing a lot of research of juice clarification, as Wei Juan etc., polygalacturonase is to the experimental study of precocious strains clarification of juice, and Anhui agronomy is circulated a notice of; And for example Zhao Xin, Lin Pan, compositional biological enzyme is to the research of flax rove treatment process.But the object of these researchs is not to prepare galacturonic acid oligosaccharides.Qian Lili is in its master thesis (fermentation of pectin lyase and the application at the unsaturated galacturonic acid oligosaccharides of preparation thereof, Zhejiang Polytechnical University's master thesis, in June, 2007) in report and utilize pectin lyase to prepare galacturonic acid oligosaccharides, but the utilising efficiency of its enzyme is not high, after the enzyme of every 100U can only process 2g pectin and enzymolysis for 24 hours, the low sugar of the polymerization degree such as monose and disaccharides could not be obtained.
Galacturonic acid is a monosaccharide units of pectin composition, does not still utilize enzyme process to obtain the method for this monomer at present.Pectin is as a kind of polysaccharide, general polygalacturonase hydrolysis of pectin obtains galacturonic acid oligosaccharides (2-10 monose), as Huilin Wang, Characterization and high-level expression of a metagenome-derived alkalinepectate lyase in recombinant Escherichia coli, Process Biochemistry, 2014, utilize polygalacturonase hydrolysate to be that disaccharides is to tetrose.If obtain monose, still needing will utilize disaccharide-hydrolysing enzymes to be hydrolyzed further could to obtain monose.The concept of oligose is put forward by B.Helgerich etc. nineteen thirty, is the low polymerization sugar of straight or branched be made up of 2 ~ 10 monose.It is the organic compound that occurring in nature one class has multiple special biological function, and distribution is extensive especially, and of a great variety, in glycobiology function, occupy sizable proportion.Oligose comprises two large classes, i.e. natural oligose and functional oligose, galacturonic acid oligosaccharides belongs to functional oligose, because lack the enzyme system of decomposition function oligose in human gastrointestinal tract inside, simultaneously it can not by human body acid, gastric enzyme degrade, directly can enter in large intestine and preferentially be utilized by bifidus bacillus, be the factor of the propagation bifidus bacillus of excellent property.Functional oligose also has some characteristics of carbohydrate, and alternative sugar material is as the seasoning matter of sweet food.Galacturonic acid oligosaccharides has promotion intestinal bifidobacteria and breeds in a large number, coordinate plant growth is grown, anticancer, reduce cholesterol, suppress the effects such as fatty liver (Qian Lili, the fermentation of pectin lyase and the application at the unsaturated galacturonic acid oligosaccharides of preparation thereof, Zhejiang Polytechnical University's master thesis, in June, 2007), therefore, its application potential is huge.
The present invention utilizes bacterial strain GS115/pel168s-9k involved in patent (patent No. ZL201110379664.X) " a kind of alkaline pectase pel168s Nucleotide majorizing sequence and high expression thereof ", carry out enzymatic production, and enzymatic hydrolysis condition is carried out to food grade pectin grope, explore the condition preparing monose and galacturonic acid oligosaccharides.
Summary of the invention
The polygalacturonase that the object of the invention is to propose to utilize pichia spp to ferment and produce has high enzyme and lives, and can the feature of depolymerized pectin rapidly and efficiently, provides a kind of method preparing monose and galacturonic acid oligosaccharides.
In order to realize object of the present invention, contriver is studied and persistent exploration by lot of experiments, finally obtains following technical scheme:
Utilize polygalacturonase cracking height gamma value pectin to prepare a method for monose and galacturonic acid oligosaccharides, the method comprises the steps:
(1) fermentation culture GS115/pel168s-9k bacterial classification prepares pectase liquid;
(2) pectin 2g being greater than 70% gamma value is dissolved in the polygalacturonase damping fluid of 100ml pH9.4, regulate pH to 8.8-9.2, after water-bath is preheated to 50 DEG C, add the pectase liquid of step (1) preparation, enzyme 100-120U alive, under 50 DEG C of condition, react 2h, constantly stir in reaction process, and in front 10min after the reaction in every 5min supplement 4g pectin, by regulating the pH=8.8-9.2 made in whole reaction, obtain enzymolysis solution;
(3) the enzymolysis solution centrifugal 10min under 3000rpm condition step (2) prepared, get supernatant liquor, in the loft drier of 65 DEG C, evaporation drying obtains galacturonic acid oligosaccharides.
Preferably, utilize polygalacturonase cracking pectin to prepare the method for monose and galacturonic acid oligosaccharides as above, wherein the polygalacturonase damping fluid described in step (2) is prepared as follows: take CaCl
20.021g, glycine 1.08775g, NaOH0.35g, with after deionized water dissolving and constant volume to 500ml.
Further preferably, polygalacturonase cracking pectin is utilized to prepare the method for monose and galacturonic acid oligosaccharides as above, wherein step (1) by activation GS115/pel168s-9k strain inoculation in liquid B MGY substratum, at 28 DEG C, shake-flask culture 2d under the condition of 220rpm, the bacterium liquid fermented is proceeded in sterile centrifugation tube, the centrifugal 10min of 3000rpm, outwell supernatant, the sterilized water accounting for centrifuge tube volume 1/3rd is added in centrifuge tube, the thalline of piping and druming at the bottom of centrifuge tube makes it resuspended, the centrifugal 10min of 3000rpm, outwell supernatant, the liquid B MMY substratum accounting for centrifuge tube volume 1/3rd is added in centrifuge tube, the thalline of piping and druming at the bottom of centrifuge tube makes it resuspended, then bacteria suspension is proceeded in liquid B MMY substratum, adding methyl alcohol to final concentration is again 1%, at 28 DEG C, shake-flask culture 4d under the condition of 220rpm, between incubation period, it is 1% that timing every day adds methyl alcohol to final concentration, cultivate proceeded in centrifuge tube by bacterium liquid time the 5th day, the centrifugal 10min of 3000rpm, get supernatant, be pectase liquid.
Processing step of the present invention is the processes such as the fermentation of polygalacturonase, the enzymolysis of substrate, centrifugal, product analysis.Concrete technology step is as follows:
1, by GS115/pel168s-9k bacterial classification at solid YPD (glucose 2.0g, peptone 2.0g, yeast extract 1.0g, agar 1.5g, distilled water 100ml) flat lining out, make it activate, from above-mentioned YPD flat board, picking individual colonies is inoculated in 250mL Erlenmeyer flask, bottle includes 50ml liquid B MGY (yeast extract 1.0g peptone 2.0g, yeast nitrogen (YNB) 0.34g, ammonium sulfate 1.0g, pH6.0 phosphoric acid buffer 10ml, glycerine 10ml, water 100ml) substratum, put into shaking table, at 28 DEG C, 2d is cultivated under the condition of 220rpm, the bacterium liquid fermented proceed in sterilized sterile centrifugation tube by aseptic operating platform, post sealing tape, trim, centrifugal 10min under 3000rpm condition in centrifuges, supernatant after centrifugal is outwelled by aseptic operating platform, the sterilized water accounting for centrifuge tube volume 1/3rd is added in centrifuge tube, to blow off the thalline at the bottom of core barrel with rifle, make it resuspended, post sealing tape, trim, centrifugal 10min under 3000rpm condition in centrifuges, supernatant after centrifugal is outwelled by aseptic operating platform, liquid B MMY (the yeast extract 1.0g accounting for centrifuge tube volume 1/3rd is added in centrifuge tube, peptone 2.0g, yeast nitrogen (YNB) 0.34g, ammonium sulfate 1.0g, pH6.0 phosphoric acid buffer 10ml, water 100ml) substratum, to blow off the thalline at the bottom of core barrel with rifle, make it resuspended, then bacteria suspension is settled to 50ml's with liquid B MMY substratum in 250ml Erlenmeyer flask, add 500 μ l methyl alcohol again, put into shaking table, at 28 DEG C, 4d is cultivated under the condition of 220rpm, between incubation period, every day, timing added 500 μ l methyl alcohol.Cultivate proceeded in centrifuge tube by bacterium liquid time the 5th day, and trim, centrifugal 10min under 3000rpm condition, gets supernatant in centrifuges, and this is enzyme liquid, puts into 4 DEG C of Refrigerator stores for subsequent use.
2, polygalacturonase enzyme activity determination.Reaction system comprises thick enzyme diluent and gets 20 μ l, adds 0.2% polygalacturonic acid solution 2ml and starts enzymatic reaction: reaction conditions is 50 DEG C of reaction 15min, by the phosphoric acid termination reaction of 3ml 0.03mol/L, surveys its absorbance by microplate reader at 235nm place.Blank is the enzyme liquid reaction (that is: first add 3ml phosphoric acid and enzyme liquid to be measured mixes, then add substrate reactions 15min) of non-activity.Get enzyme liquid, survey its enzyme by A235 method and live.
In formula: 4600 (Lmol
-1cm
-1the molar absorptivity of)-unsaturated galacturonic acid at 235nm place
T (min)-time of enzymatic reacting (in the linearity range of enzymatic reaction)
B (cm)-cuvette thickness
Simplify: enzyme (U/ml)=3.6232* extension rate alive * OD
235
Result: enzyme (U/ml)=3.6232* extension rate alive * OD
235=3.6232*100* (0.538-0.277)=110.87U/ml
3, enzyme digestion reaction.Reaction system is pectin 10g, enzyme liquid 1ml, pH9.0, reaction times 2h.Concrete operations are the damping fluid 2 grams of pectin being dissolved in 100ml pH9.4 (is CaCl
20.021g, glycine 1.08775g, NaOH0.35g, with 500ml deionized water constant volume to 500ml) in, about pH to 9 is regulated with NaOH (5M), preheating 10min in 50 DEG C of water-baths, add 1ml enzyme liquid (that is: every 100ml reaction solution 110U polygalacturonase), react under 50 DEG C of conditions, reaction times is 2h, constantly stir with glass stick in reaction process, in 10min after this, add 4.0g substrate every 5min and regulate about pH to 9 with NaOH (5M), in reaction process after this, every 5min also will regulate a pH, pH in whole reaction process is made to maintain about 9.
4, by reaction solution centrifugal 10min under 3000rpm condition, get supernatant liquor and carry out liquid chromatography mass spectrometric detection, mass spectrograph used is Agilent1260-6224LC-ESI-TOF-MS.The results are shown in Figure 1, pectin obtains monose and the disaccharides oligosaccharides to seven sugar after enzymolysis, wherein based on monose to pentasaccharides.
5, get centrifugal after precipitation, measure the percent hydrolysis of pectin.The precipitation obtained after enzymolysis solution is centrifugal is placed on and is dried to its weight in 65 DEG C of loft drier and no longer reduces together with centrifuge tube, and after deducting the weight of centrifuge tube, the percent hydrolysis calculating pectin reaches 93%.
Compared with prior art, the method that the present invention utilizes polygalacturonase cracking pectin to prepare monose and galacturonic acid oligosaccharides has the advantage that technique is simple, cost is low, the utilising efficiency of enzyme is high, the percent hydrolysis of pectin can up to more than 90% simultaneously, and hydrolysate contains a large amount of monose.
Accompanying drawing explanation
Fig. 1 be embodiment five centrifugal after enzymolysis solution do liquid chromatography mass spectrometric detect result.
Specific embodiment
The bacterial strain that following examples adopt is GS115/pel168s-9k, the pel gene transformation of codon optimization obtains by this bacterial strain in pichia spp genome, specifically see patent ZL201110379664.X " a kind of alkaline pectase pel168s Nucleotide majorizing sequence and high expression thereof ".This bacterial strain can great expression polygalacturonase be discharged to outside born of the same parents under suitable fermentation condition, makes nutrient solution have very high enzyme concn.The substrate that the present invention adopts is the high fat rapid hardening pectin 0121, viscosity 400-500mpa.s, gamma value >70% purchased from Anhui Xin Chuan chemical industry.
Embodiment one: the fermentation of polygalacturonase
Prepare 50mlBMGY liquid nutrient medium (yeast extract 0.5g peptone 1.0g respectively, yeast nitrogen (YNB) 0.27g, ammonium sulfate 0.5g, pH6.0 phosphoric acid buffer 5ml, glycerine 5ml, water 50ml) and 50mlBMMY liquid nutrient medium (yeast extract 0.5g, peptone 1.0g, yeast nitrogen (YNB) 0.27g, ammonium sulfate 0.5g, pH6.0 phosphoric acid buffer 5ml, water 50ml), 121 DEG C of sterilizings, preparation 100ml solid YPD substratum (glucose 2.0g, peptone 2.0g, yeast extract 1.0g, agar 1.5g, distilled water 100ml), plate is down flat after 108 DEG C of sterilizings.By the GS115/pel168s-9k bacterial classification of preservation at the flat lining out of solid YPD, it is made to activate, from above-mentioned YPD flat board, picking individual colonies is inoculated in 250mL Erlenmeyer flask, bottle includes 50ml liquid B MGY substratum, put into shaking table, at 28 DEG C, 2d is cultivated under the condition of 220rpm, the bacterium liquid fermented proceed in sterilized sterile centrifugation tube by aseptic operating platform, centrifugal 10min under 3000rpm condition, supernatant after centrifugal is outwelled by aseptic operating platform, resuspended with the sterilized water accounting for centrifuge tube volume 1/3rd, post sealing tape, trim, centrifugal 10min under 3000rpm condition, supernatant after centrifugal is outwelled by aseptic operating platform, with the resuspended thalline of liquid B MMY substratum accounting for centrifuge tube volume 1/3rd, then bacteria suspension is proceeded in 250ml Erlenmeyer flask, 50ml is settled to liquid B MMY substratum, add 500 μ l methyl alcohol again, put into shaking table, at 28 DEG C, 4d is cultivated under the condition of 220rpm, between incubation period, every day, timing added 500 μ l methyl alcohol.Cultivate proceeded in centrifuge tube by bacterium liquid time the 5th day, and trim, centrifugal 10min under 3000rpm condition, gets supernatant in centrifuges, and this is enzyme liquid, and recording the work of its enzyme by A235 method is 110.87U/ml, puts into 4 DEG C of Refrigerator stores for subsequent use.
The enzymolysis of the 330U polygalacturonase of embodiment two: 100ml substrate
2.0g substrate is dissolved in the damping fluid of 100mlpH9.4, about pH to 9 is regulated with dense NaOH, preheating in 50 DEG C of water-baths, add enzyme liquid (that is: every 10g pectin 330U polygalacturonase) prepared by 3ml embodiment one, react under 50 DEG C of conditions, reaction times is 2h, constantly stir with glass stick in reaction process, in 10min after this, add 4.0g substrate every 5min and regulate about pH to 9 with NaOH (5M), in reaction process after this, every 5min also will regulate a pH, makes pH in whole reaction process maintain about 9.After result enzymolysis 2h, enzymolysis solution good fluidity, recording hydrolyzed pectin rate is 93%.
The enzymolysis of the 11U polygalacturonase of embodiment three: 100ml substrate
2.0g substrate is dissolved in the polygalacturonase damping fluid of 100mlpH9.4, about pH to 9 is regulated with NaOH (5M), preheating in 50 DEG C of water-baths, add the enzyme liquid (that is: every 10g pectin 11U polygalacturonase) of 100 μ l embodiment one preparations, react under 50 DEG C of conditions, reaction times is 2h, constantly stir with glass stick in reaction process, in 10min after this, often cross 5min add 4.0g substrate and regulate about pH to 9 with NaOH (5M), in reaction process after this, every 5min also will regulate a pH, makes pH in whole reaction process maintain about 9.The pectin added after result 2h does not dissolve completely, enzymolysis solution thickness, the incomplete enzymolysis of substrate.
The enzymolysis of the 55U polygalacturonase of embodiment four: 100ml substrate
2.0g substrate is dissolved in the polygalacturonase damping fluid of 100mlpH9.4, about pH to 9 is regulated with NaOH (5M), preheating in 50 DEG C of water-baths, add the enzyme liquid (that is: every 10g pectin 55U polygalacturonase) of 500 μ l embodiment one preparations, react under 50 DEG C of conditions, reaction times is 2h, constantly stir with glass stick in reaction process, in 10min after this, often cross 5min add 4.0g substrate and regulate about pH to 9 with NaOH (5M), in reaction process after this, every 5min also will regulate a pH, makes pH in whole reaction process maintain about 9.Enzymolysis solution still thickness after result 2h, substrate enzymolysis is not thorough.
The enzymolysis of the 110U polygalacturonase of embodiment five: 100ml substrate
2.0g substrate is dissolved in the polygalacturonase damping fluid of 100mlpH9.4, about pH to 9 is regulated with NaOH (5M), preheating in 50 DEG C of water-baths, add enzyme liquid (that is: every 10g pectin 110U polygalacturonase) prepared by 1ml embodiment one, react under 50 DEG C of conditions, reaction times is 2h, constantly stir with glass stick in reaction process, in 10min after this, often cross 5min add 4.0g substrate and regulate about pH to 9 with NaOH (5M), in reaction process after this, every 5min also will regulate a pH, makes pH in whole reaction process maintain about 9.Enzymolysis solution good fluidity after result 2h, the percent hydrolysis recording pectin is 93%.Compare with embodiment two, hydrolyzed state and percent hydrolysis be obviously difference not, illustrates that the enzyme of 110U can be hydrolyzed 10g substrate.
The method of calculation of hydrolyzed pectin rate are as follows:
1, the precipitation obtained after enzymolysis solution is centrifugal is placed in 65 DEG C of loft drier and is dried to its weight and no longer reduces its weight m of postscript together with centrifuge tube
1.
2, with clear water by the precipitation wash clean in centrifuge tube, then clean centrifuge tube is placed in 65 DEG C of loft drier and is dried to its weight and no longer reduces its weight m of postscript
2.
Percent hydrolysis measurement result: (such as m
1=25.93, m
2=25.19)
Percent hydrolysis=[10-(m
1-m
2)]/10*100%=[10-(25.93-25.19)]/10*100%.
Embodiment six: the detection of hydrolysate
Repeat embodiment 4 hydrolyzable moiety, by reaction solution centrifugal 10min under 3000rpm condition, get supernatant liquor and carry out liquid chromatography mass spectrometric detection, mass spectrograph used is Agilent1260-6224LC-ESI-TOF-MS.The results are shown in Figure 1, pectin obtain after enzymolysis monose to and disaccharides to the oligosaccharides of seven sugar, wherein based on monose to pentasaccharides, monose peak value reaches 10
5, disaccharides reaches 10 to pentasaccharides
4.
Claims (3)
1. utilize polygalacturonase cracking height gamma value pectin to prepare a method for monose and galacturonic acid oligosaccharides, it is characterized in that the method comprises the steps:
(1) fermentation culture GS115/pel168s-9k bacterial classification prepares pectase liquid;
(2) 2g pectin is dissolved in the polygalacturonase damping fluid of 100ml pH9.4, regulate pH to 8.8-9.2, after water-bath is preheated to 50 DEG C, add the pectase liquid of enzyme 100-120U alive prepared by step (1), under 50 DEG C of condition, react 2h, constantly stir in reaction process, and in front 10min after the reaction in every 5min supplement 4g pectin, by regulating the pH=8.8-9.2 made in whole reaction, obtain enzymolysis solution;
(3) the enzymolysis solution centrifugal 10min under 3000rpm condition step (2) prepared, gets supernatant liquor, and evaporation oven dry obtains monose and galacturonic acid oligosaccharides solid 0.74 gram, and the percent hydrolysis of pectin reaches 93%.
2. the method utilizing polygalacturonase cracking height gamma value pectin to prepare monose and galacturonic acid oligosaccharides according to claim 1, enzymolysis solution is centrifugal 10min under 3000rpm condition, get supernatant liquor and carry out liquid chromatography mass spectrometric detection, display hydrolysate is monose and the disaccharides galacturonic acid oligosaccharides to seven sugar, wherein based on monose to pentasaccharides, monose peak value reaches 10
5, disaccharides reaches 10 to pentasaccharides
4.
3.GS115/pel168s-9k bacterial classification can be hydrolyzed high gamma value pectin and the application produced in the polygalacturonase of monose and galacturonic acid oligosaccharides in preparation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510037006.0A CN104593450B (en) | 2015-01-26 | 2015-01-26 | A kind of method for preparing monose and galacturonic acid oligosaccharides using pectin enzymatic lysis high ester degree pectin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510037006.0A CN104593450B (en) | 2015-01-26 | 2015-01-26 | A kind of method for preparing monose and galacturonic acid oligosaccharides using pectin enzymatic lysis high ester degree pectin |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104593450A true CN104593450A (en) | 2015-05-06 |
| CN104593450B CN104593450B (en) | 2018-04-27 |
Family
ID=53119503
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510037006.0A Active CN104593450B (en) | 2015-01-26 | 2015-01-26 | A kind of method for preparing monose and galacturonic acid oligosaccharides using pectin enzymatic lysis high ester degree pectin |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104593450B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105647993A (en) * | 2016-02-24 | 2016-06-08 | 安徽宇宁生物科技有限公司 | Production technology of liquid oligogalacturonic acid pectin |
| CN108929889A (en) * | 2018-08-01 | 2018-12-04 | 南京林业大学 | A method of preparing pectin disaccharides and pectin trisaccharide |
| WO2023036219A1 (en) * | 2021-09-08 | 2023-03-16 | 上海见杉医学技术有限公司 | Homogalacturonan, complex, and preparation method therefor and use thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102604977A (en) * | 2011-11-22 | 2012-07-25 | 湖北大学 | Optimized nucleotide sequence of alkaline pectinase pell68s and high-level expression method thereof |
| CN103445160A (en) * | 2013-08-11 | 2013-12-18 | 吉林农业大学 | Method for preparing ginseng pulp through enzymolysis of pectinase and ginseng pulp |
-
2015
- 2015-01-26 CN CN201510037006.0A patent/CN104593450B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102604977A (en) * | 2011-11-22 | 2012-07-25 | 湖北大学 | Optimized nucleotide sequence of alkaline pectinase pell68s and high-level expression method thereof |
| CN103445160A (en) * | 2013-08-11 | 2013-12-18 | 吉林农业大学 | Method for preparing ginseng pulp through enzymolysis of pectinase and ginseng pulp |
Non-Patent Citations (1)
| Title |
|---|
| 钱丽丽: "果胶裂解酶的发酵及其在制备不饱和低聚糖方面的应用", 《中国优秀硕士学位论文全文数据库（工程科技I辑）》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105647993A (en) * | 2016-02-24 | 2016-06-08 | 安徽宇宁生物科技有限公司 | Production technology of liquid oligogalacturonic acid pectin |
| CN108929889A (en) * | 2018-08-01 | 2018-12-04 | 南京林业大学 | A method of preparing pectin disaccharides and pectin trisaccharide |
| CN108929889B (en) * | 2018-08-01 | 2021-09-28 | 南京林业大学 | Method for preparing pectin disaccharide and pectin trisaccharide |
| WO2023036219A1 (en) * | 2021-09-08 | 2023-03-16 | 上海见杉医学技术有限公司 | Homogalacturonan, complex, and preparation method therefor and use thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104593450B (en) | 2018-04-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Chen et al. | New insight into bamboo shoot (Chimonobambusa quadrangularis) polysaccharides: Impact of extraction processes on its prebiotic activity | |
| CN105087456B (en) | A method of producing the recombined bacillus subtilis building of specified molecular weight hyaluronic acid | |
| CN108118003B (en) | Black yeast, culture medium and method for producing beta-glucan, and black yeast culture and composition containing beta-glucan | |
| Sitanggang et al. | Aspects of glucosamine production using microorganisms | |
| CN101360829A (en) | Fermentation and culture method, fermented plant extract, composition containing fermented plant extract, method for producing lipopolysaccharide and lipopolysaccharide | |
| CN105123929B (en) | A kind of brown alga oligose dairy products reducing cholesterol | |
| Liu et al. | Isolation and characterization of curdlan produced by Agrobacterium HX1126 using α-lactose as substrate | |
| CN110172486A (en) | A method of synthesis 2'-Fucosyl lactose | |
| CN105602968A (en) | Esterase and application thereof | |
| CN104593450A (en) | Method for splitting pectin with high esterification degree to prepare monosaccharide and oligogalacturonic acid employing pectinase | |
| CN109609422A (en) | One kind can produce the construction method of 2 '-rock algae base lactose bacterial strains | |
| CN102690795B (en) | Trehalose synthase of streptomyces griseochromogenes and coding gene and application of trehalose synthase | |
| CN113265337B (en) | Marine aspergillus versicolor and isolated culture method and application thereof | |
| US20080064073A1 (en) | Process for Preparing | |
| TWI712689B (en) | CULTURING MEDIUM AND METHOD FOR PRODUCING β-GLUCAN, A CULTURE OF AUREOBASIDIUM PULLULANS AND A COMPOSITION COMPRISING THE SAME | |
| CN102120999A (en) | Method for synthesizing human milk fucosylation oligosaccharide by using genetic engineering strain through coupling and fermenting | |
| Xu et al. | A new strategy to improve Ganoderma polysaccharides production by symbiotic fungi elicitors through activating the biosynthetic pathway | |
| CN103045561B (en) | Solid state fermentation production method for beta-D-fructofuranosidase | |
| CN112553272B (en) | Method for improving hyaluronic acid yield | |
| CN102154360B (en) | Recombinant expression vectors pQHK and pHK producing hyaluronic acid and construction method thereof | |
| EP1266968B1 (en) | Microorganism producing fructosyl transferase and method for producing fructooligosaccharides and neofructooligosaccharides using the same | |
| CN106755197A (en) | A kind of method that utilization linear maltooligosacchaeides generation enzyme prepares straight chain MALTOHAXAOASE | |
| CN106282142A (en) | The preparation method of the α tilactase that a kind of β mannosidase content is low | |
| CN107058266B (en) | A method of lysozyme is prepared by zymotechnique | |
| CN105602969A (en) | Esterase and application thereof |
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 |