CN1211622A - Method for continuous preparation of isomalt oligose by use of hollow fiber enzyme membrane reactor - Google Patents
Method for continuous preparation of isomalt oligose by use of hollow fiber enzyme membrane reactor Download PDFInfo
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- CN1211622A CN1211622A CN 98111000 CN98111000A CN1211622A CN 1211622 A CN1211622 A CN 1211622A CN 98111000 CN98111000 CN 98111000 CN 98111000 A CN98111000 A CN 98111000A CN 1211622 A CN1211622 A CN 1211622A
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Abstract
A process for continuously preparing isomaltoligose (in the form of syrup or powder) with hollow cellulase membrane reactor features that starch, especially the starch of cassava or corn, is used as raw material, and continuous process with hollow cellulase membrane reactor is used, which includes saccharifying with alpha-amylase and isoamylase (or other) to prepare maltoligose, and transforming glycoside with alpha-glucosiduronic acid and fungus amylase to obtain isomaltoligose. Its advantages are shorter reaction period, less consumption of enzyme, stable quality of product and high content of branched saccharide (more than 60%).
Description
The present invention relates to a kind of manufacture method of oligose, exactly, it is the coming of new technology of dextrinosan.
Dextrinosan claims isomalto Oligosaccharide (Isomaltooligosaccharide) again or is ramiform oligose (Heterooligosaccharide), be meant that intramolecularly contains α-1,6 glycosidic links, mainly containing glucose polymerization degree is the ramiform oligose of 2-5, contains glucose and a small amount of straight chain malto-oligosaccharide in addition.It is a kind of new type functional oligose, is the bifidus bacillus multiplicaiton factor, has anti-dental caries, prevents constipation, premium properties such as heat-resisting, acidproof, moisture retention, water-activity are low.The former biochemical scientific research institution's exploitation alpha-glucosidase of 82 years Japanese woodss, the clear and industry company of 85 years Japan is with IMO-500, and IMO-900P introduces to the market.By retrieval as can be known: Jin Qirong, Xu Qin are published in " starch and Dian Fentang " 1996 (3) 5-9 pages or leaves, and name is called in " dextrinosan (oligosaccharides) character and production and application " literary composition to be introduced; It is to be raw material with starch, and starch slurry is through α-Dian Fenmeiyehua (DE6-10); Beta-amylase (or also adding Pullulanase), alpha-glucosaccharase enzyme glycolysis change glycosides: filtration, decolouring, desalination, vacuum concentration get IMO-500.It is a kind of intermittence production technique, i.e. saccharification, commentaries on classics glycosides carry out in a retort simultaneously, owing to have the product inhibition problem, thereby long reaction time (36-48h), enzyme dosage big (every jar all will add necessary enzyme).
The present invention seeks to adopt the two enzyme membrane serialization systems of tubular fibre enzyme mebrane reactor, with starch is raw material, produce the intermediate product malto-oligosaccharide with α-Dian Fenmei and Pullulanase (or isoamylase) saccharification, the back is changeed glycosides with alpha-glucosidase and fungal amylase and is made the dextrinosan the finished product.
The present invention realizes by following technical proposals, and a kind of tubular fibre enzyme mebrane reactor system is provided, and makes malto-oligosaccharide with continuous conversion; After change glycosides more continuously and obtain dextrinosan.Make saccharification, change glycosides optimal control respectively, enzyme two enzyme membrane continuous conversions of recycle repeatedly changes glycosides novel process (claiming malto-oligosaccharide to change the glycosides method).(a) the coming of new technology of dextrinosan (syrup, powder), it adopts the two enzyme membrane serialization systems of tubular fibre enzyme mebrane reactor, adopt the polysulfone hollow fibre ultra-fine filter, molecular weight cut-off 10000, cytochrome C rejection 100%, (b) its operational path: with starch is raw material (particularly tapioca (flour) or W-Gum), prepare malto-oligosaccharide with α-Dian Fenmei and Pullulanase (or isoamylase) saccharification, the back is changeed glycosides with alpha-glucosidase and fungal amylase and is made dextrinosan.
The tubular fibre enzyme mebrane reactor continuous process manufacture method of dextrinosan, the selection of polysulfone hollow fibre ultra-fine filter molecular weight cut-off, construct singularity and practice summary from carbohydrate molecule: the selection of molecular weight cut-off is that the carbohydrate molecule amount is taken advantage of 10 times, carbohydrate molecule is filtered smoothly, as MALTOHAXAOASE or ramiform MALTOHAXAOASE molecular weight is 972, and its molecular weight cut-off should select 9720 to approximate 10000.
Its method of manufacturing technology is:
Saccharification: two jars of liquefiers liquefy respectively earlier.One jar to insert storage tank standby, inserts 1 for one jar
#Retort is regulated pH5.3-5.8 with 5N HCl, add α-Dian Fenmei (2.0-2.5 μ/g starch), isoamylase (10.0-13.0 μ/g starch) or Pullulanase (2-4L/g starch) stir saccharification 1.5h in 45-50 ℃, after begin circulation, stablize peristaltic pump pressure 0.1Mpa/cm
2, circulate after half an hour, from storage tank and the moisturizing of replenisher tank continuous feeding, mend enzyme behind the saccharification 3h, α-Dian Fenmei is added 15-20%, and isoamylase adds 5%, and mending the enzyme running time is 1.0-1.5h (but Continuous Flow adds or adds in batches).Maintenance saccharification liquid and out-feed liquid pol all about 25, draw product may command DE value and deposit between the 29-31, and the product sampling is HPLC and is detected the malto-oligosaccharide component concentration.
Change glycosides: the said products (about 25% malto-oligosaccharide solution) injects 2
#In the retort, regulate pH5.0-5.5, add alpha-glucosidase (300-1500 μ/g malto-oligosaccharide), fungal amylase (60-100 μ/g malto-oligosaccharide), in 55-58 ℃ of stirring reaction 6h, (analyse maltose in the detecting reactant with ply of paper, trisaccharide maltose sugar spot disappears substantially), be the enzyme membrane reaction cycle, from storage tank feed supplement and moisturizing, feed supplement and discharging are in a basic balance, mend enzyme (but stream adds or adding in batches), alpha-glucosidase adds 20%, and fungal amylase is added 5-10%, keeps reaction solution and out-feed liquid pol all about 25, the control reactant residence time is 3-4h, and the product sampling is IIPLC and is detected the dextrinosan component concentration.
Saccharification of the present invention, commentaries on classics glycosides are able to the continuous production technology of optimal control respectively.
The present invention is to be raw material, particularly tapioca (flour) or W-Gum with starch.
This pair of enzyme membrane continuous processing of the present invention, make saccharification, commentaries on classics glycosides be able to optimal control respectively, experimental result shows the reaction time that shortens saccharification greatly, change glycosides (being the 1/6-1/4 8-9h only of original 36-48h), enzyme dosage is economized (with continuous three batches of calculating, enzyme can save 53%), constant product quality, DP2-5 branched sugar content is greater than 60%, surpass the quality index (52%) of Japanese like product [IMO-500 or Barrow Lay husband (バ ノ ラ Star プ)], the present invention also is a kind of novel method of enzyme immobilization.
Accompanying drawing is an of the present invention pair of enzyme membrane reaction process schema
Mashing process process: liquefying starch (liquefier) is put into the 1# reactive tank, pass through then peristaltic pump Enter the 1# ultrafilter, through returning the 1# reactive tank after the 1# ultrafiltration, the product of separation flows into the 2# reactive tank. Many dotted lines show that its raw material can be intermittent production in the former stockline that flows into the 2# reactive tank, Also can be that the serialization of enzyme membrane reaction process is produced.
Turn to the glycosides technical process: the reactant liquor in the 2# reactive tank is entered the 2# ultrafilter by peristaltic pump, through 2# Return behind the ultrafilter in the 2# reactive tank, products of separated flows in the product-collecting groove through desalination, makes different Fructus Hordei Germinatus Compound sugar.
Below the present invention is described in further detail by specific embodiment.
Embodiment 1: tapioca (flour) is that raw material prepares malto-oligosaccharide continuously.
100 portions of tapioca (flour)s and 200 parts of water are made into starch slurry, regulate pH6.0-6.5, add 5%CaCl with 5% sodium carbonate solution
21-2ml places 85 ℃ of thermostat water bath heating, stirs, before slurry temperature rises to starch pasting, during proper temperature, add the liquefaction of α-Dian Fenmei (enzyme activity 0.5-0.8 μ/g starch) gelatinization limit, limit immediately, continue to be warming up to 70+2 ℃, constant temperature 10-15min, liquefaction finishes.Sampling 10ml surveys its DE value and is 5-6, and same quadrat method liquefies second part, the 3rd part ... as saccharification feed supplement liquid.
One jar of liquefier is inserted 1# retort (another jar put into storage tank as feed supplement liquid), Dropwise 5 NHCl, regulate pH5.3～5.8, add α-Dian Fenmei (enzyme activity 2.0～2.5 μ/g starch), isoamylase (10.0～13.0 μ/g starch) begins circulation behind 45～50 ℃ of stirrings, saccharification 1.5h, stablize peristaltic pump pressure 0.1Mpa/cm
2The circulation half an hour after continuous feeding and moisturizing, behind the saccharification 3h, (α-Dian Fenmei adds 15～20% to mend enzyme, isoamylase adds 5%), mending the enzyme running time is 1.0～1.5h, and (but Continuous Flow adds or adds in batches), the pol that keeps saccharification liquid and product is all about 25, draw product may command DE value between 29～31, the product sampling is HPLC and is detected the malto-oligosaccharide component concentration.
Embodiment 2: be raw material with the W-Gum, prepare malto-oligosaccharide continuously.
100 parts of W-Gums and 200 parts of water are made into starch slurry, regulate pH6.0-6.5, add 5%CaCl with 5% sodium carbonate solution
21-2ml places 90 ℃ of thermostat water bath heating, stirs, before slurry temperature rises to starch pasting, during proper temperature, add the liquefaction of α-Dian Fenmei (enzyme activity 0.5-0.8 μ/g starch) gelatinization limit, limit immediately, continue to be warming up to 80 ± 2 ℃, constant temperature 10-15min, liquefaction finishes.Sampling 10ml surveys its DE value and is 5-6, and same quadrat method liquefies second part, the 3rd part ... as saccharification feed supplement liquid.
One jar of liquefier is inserted 1# retort (another jar put into storage tank as feed supplement liquid), Dropwise 5 NHCl, regulate pH5.3～5.8, add α-Dian Fenmei (enzyme activity 2.0～2.5 μ/g starch), isoamylase (10.0～13.0 μ/g starch) begins circulation behind 45～50 ℃ of stirrings, saccharification 1.5h, stablize peristaltic pump pressure 0.1Mpa/cm
2The circulation half an hour after continuous feeding and moisturizing, behind the saccharification 3h, (α-Dian Fenmei adds 15～20% to mend enzyme, isoamylase adds 5%), mending the enzyme running time is 1.0～1.5h, and (but Continuous Flow adds or adds in batches), the pol that keeps saccharification liquid and product is all about 25, draw product may command DE value between 29～31, the product sampling is HPLC and is detected the malto-oligosaccharide component concentration.
Embodiment 3: with the cassava malto-oligosaccharide is that raw material prepares dextrinosan continuously.
Above-mentioned about 25% cassava malto-oligosaccharide solution adds 2
#In the retort, regulate pH5.0～5.5, add alpha-glucosidase (enzyme activity 300～1500 μ/g malto-oligosaccharide), fungal amylase (60～100 μ/g malto-oligosaccharide), in 55～58 ℃ of stirring reaction 6h, (analysing maltose in the detecting reactant, trisaccharide maltose sugar spot with ply of paper disappears substantially), beginning enzyme membrane reaction cycle, the control of feed supplement discharging speed is in a basic balance, keep saccharification liquid, out-feed liquid pol about 25, making reactant residence time is 3～4h, the product sampling is made HPLC and is detected the dextrinosan component concentration.
Embodiment 4: with the malted maize oligose is raw material, prepares dextrinosan continuously.
With 25% malted maize oligosaccharide solution, add 2
#In the retort, regulate p II 5.0～5.5, add alpha-glucosidase (enzyme activity 300～1500 μ/g malto-oligosaccharide), fungal amylase (80～120 μ/g malto-oligosaccharide), in 55～58 ℃ of stirring reaction 6h, (analysing maltose in the detecting reactant, trisaccharide maltose sugar spot with ply of paper disappears substantially), beginning enzyme membrane reaction cycle, the control of feed supplement discharging speed is in a basic balance, keep saccharification liquid and out-feed liquid pol equal about 25, making reactant residence time is 3～4h, and the product sampling is made HPLC and detected the dextrinosan component concentration.
Claims (6)
1. a tubular fibre enzyme mebrane reactor prepares the method for dextrinosan continuously, it is characterized in that;
(a) it adopts the two enzyme membrane serialization systems of tubular fibre enzyme mebrane reactor, adopts the polysulfone hollow fibre ultra-fine filter, molecular weight cut-off 10000, cytochrome C rejection 100%;
(b) its operational path: with starch is raw material, prepares malto-oligosaccharide with α-Dian Fenmei and Pullulanase (or isoamylase) saccharification, and the back is changeed glycosides with alpha-glucosidase and fungal amylase and made dextrinosan.
2. the method for preparing dextrinosan according to the tubular fibre enzyme mebrane reactor of claim 1 continuously, the tubular fibre enzyme mebrane reactor continuous process manufacture method that it is characterized in that dextrinosan, the selection of polysulfone hollow fibre ultra-fine filter molecular weight cut-off is from carbohydrate molecule structure singularity and practice summary; The selection of molecular weight cut-off is that the carbohydrate molecule amount is taken advantage of 10 times, and carbohydrate molecule is filtered smoothly.
3. the method for preparing dextrinosan according to the tubular fibre enzyme mebrane reactor of claim 1 continuously is characterized in that saccharification is: the two jars of liquefiers that liquefy respectively earlier, one jar to insert storage tank standby, inserts 1 for one jar
#Retort is regulated pH5.3-5.8 with 5N HCl, add α-Dian Fenmei (2.0-2.5 μ/g starch), isoamylase (10.0-13.0 μ/g starch) or Pullulanase (2-4L/g starch) begin circulation behind 45-50 ℃ of stirring saccharification 1.5h, stablize peristaltic pump pressure 0.1Mpa/cm
2The circulation half an hour after, from storage tank and the moisturizing of replenisher tank continuous feeding, mend enzyme behind the saccharification 3h, α-Dian Fenmei is added 15-20%, and isoamylase adds 5%, mending the enzyme running time is 1.0-1.5h (but Continuous Flow adds or adds in batches), maintenance saccharification liquid and out-feed liquid pol all about 25, draw product may command DE value between 29-31, and the product sampling is HPLC and is detected the malto-oligosaccharide component concentration.
4. the method for preparing dextrinosan according to the tubular fibre enzyme mebrane reactor of claim 1 continuously, it is characterized in that changeing glycosides technology is that 25% malto-oligosaccharide solution injects 2
#In the retort, regulate pH5.0-5.5, add alpha-glucosidase (300-1500 μ/g malto-oligosaccharide), fungal amylase (60-100 μ/g malto-oligosaccharide), in 55-58 ℃ of stirring reaction 6h, (analyse maltose in the detecting reactant with ply of paper, trisaccharide maltose sugar spot disappears substantially), be the enzyme membrane reaction cycle, from storage tank feed supplement and moisturizing, feed supplement and discharging are in a basic balance, mend enzyme (but stream adds or adding in batches), alpha-glucosidase adds 20%, and fungal amylase is added 5-10%, keeps reaction solution and out-feed liquid pol all about 25, the control reactant residence time is 3-4h, and the product sampling is HPLC and is detected dextrinosan content.
5. the method for preparing dextrinosan according to claim 1,3 or 4 described tubular fibre enzyme mebrane reactors continuously is characterized in that saccharification, changes the continuous production technology that glycosides is able to optimal control respectively.
6. the method for preparing dextrinosan according to the described tubular fibre enzyme mebrane reactor of claim 1 continuously is characterized in that the starch material that is adopted is is raw material with tapioca (flour) or W-Gum.
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| CN98111000A CN1078615C (en) | 1998-08-06 | 1998-08-06 | Method for continuous preparation of isomalt oligose by use of hollow fiber enzyme membrane reactor |
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| CN98111000A CN1078615C (en) | 1998-08-06 | 1998-08-06 | Method for continuous preparation of isomalt oligose by use of hollow fiber enzyme membrane reactor |
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| CN1211622A true CN1211622A (en) | 1999-03-24 |
| CN1078615C CN1078615C (en) | 2002-01-30 |
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| CN100381472C (en) * | 2006-05-26 | 2008-04-16 | 陕西省微生物研究所 | Method for directly preparing malt sugar group-beta-cyclodextrin using starch |
| CN101433283B (en) * | 2008-12-31 | 2011-11-23 | 保龄宝生物股份有限公司 | Syrup for roasting and preparation method thereof |
| CN102559812A (en) * | 2012-01-20 | 2012-07-11 | 吉林农业大学 | Method for preparing maltose syrup by continuous saccharification of enzyme membrane reactor |
| CN102086234B (en) * | 2009-12-07 | 2012-10-03 | 北大国际医院集团重庆大新药业股份有限公司 | Preparation method for improving quality of medium molecular weight hydroxyethyl starch |
| CN104204216A (en) * | 2012-03-28 | 2014-12-10 | 丹尼斯科美国公司 | Method for making high maltose syrup |
| CN109690297A (en) * | 2016-07-01 | 2019-04-26 | 沃特世科技公司 | The method of the glycosylamine of label is quickly prepared from complex matrices using deglycosylation on molecular weight retention filtering and filter |
| US11035832B2 (en) | 2016-06-21 | 2021-06-15 | Waters Technologies Corporation | Methods of electrospray ionization of glycans modified with amphipathic, strongly basic moieties |
| US11371996B2 (en) | 2014-10-30 | 2022-06-28 | Waters Technologies Corporation | Methods for the rapid preparation of labeled glycosylamines and for the analysis of glycosylated biomolecules producing the same |
| US11448652B2 (en) | 2011-09-28 | 2022-09-20 | Waters Technologies Corporation | Rapid fluorescence tagging of glycans and other biomolecules with enhanced MS signals |
| US11747310B2 (en) | 2014-11-13 | 2023-09-05 | Waters Technologies Corporation | Methods for liquid chromatography calibration for rapid labeled N-glycans |
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| JP2607876B2 (en) * | 1987-03-06 | 1997-05-07 | 日本食品化工株式会社 | Method for producing high-purity isomaltose |
| JPH04112797A (en) * | 1990-08-31 | 1992-04-14 | Fuooku Ueizu Japan:Kk | Production of isomaltooligosaccharide |
| CN1116657A (en) * | 1995-04-24 | 1996-02-14 | 国内贸易部西安油脂科学研究设计院 | Process for producing soya oligose by membrane separation process |
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1998
- 1998-08-06 CN CN98111000A patent/CN1078615C/en not_active Expired - Fee Related
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| CN100381472C (en) * | 2006-05-26 | 2008-04-16 | 陕西省微生物研究所 | Method for directly preparing malt sugar group-beta-cyclodextrin using starch |
| CN101433283B (en) * | 2008-12-31 | 2011-11-23 | 保龄宝生物股份有限公司 | Syrup for roasting and preparation method thereof |
| CN102086234B (en) * | 2009-12-07 | 2012-10-03 | 北大国际医院集团重庆大新药业股份有限公司 | Preparation method for improving quality of medium molecular weight hydroxyethyl starch |
| US11448652B2 (en) | 2011-09-28 | 2022-09-20 | Waters Technologies Corporation | Rapid fluorescence tagging of glycans and other biomolecules with enhanced MS signals |
| CN102559812A (en) * | 2012-01-20 | 2012-07-11 | 吉林农业大学 | Method for preparing maltose syrup by continuous saccharification of enzyme membrane reactor |
| CN104204216A (en) * | 2012-03-28 | 2014-12-10 | 丹尼斯科美国公司 | Method for making high maltose syrup |
| US11371996B2 (en) | 2014-10-30 | 2022-06-28 | Waters Technologies Corporation | Methods for the rapid preparation of labeled glycosylamines and for the analysis of glycosylated biomolecules producing the same |
| US11747310B2 (en) | 2014-11-13 | 2023-09-05 | Waters Technologies Corporation | Methods for liquid chromatography calibration for rapid labeled N-glycans |
| US11035832B2 (en) | 2016-06-21 | 2021-06-15 | Waters Technologies Corporation | Methods of electrospray ionization of glycans modified with amphipathic, strongly basic moieties |
| CN109690297A (en) * | 2016-07-01 | 2019-04-26 | 沃特世科技公司 | The method of the glycosylamine of label is quickly prepared from complex matrices using deglycosylation on molecular weight retention filtering and filter |
| US11150248B2 (en) | 2016-07-01 | 2021-10-19 | Waters Technologies Corporation | Methods for the rapid preparation of labeled glycosylamines from complex matrices using molecular weight cut off filtration and on-filter deglycosylation |
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