CN105861595A - Method for quickly preparing galactomannan using guar gum - Google Patents
Method for quickly preparing galactomannan using guar gum Download PDFInfo
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- CN105861595A CN105861595A CN201610492584.8A CN201610492584A CN105861595A CN 105861595 A CN105861595 A CN 105861595A CN 201610492584 A CN201610492584 A CN 201610492584A CN 105861595 A CN105861595 A CN 105861595A
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- 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
- C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
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- 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
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
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Abstract
The invention discloses a method for quickly preparing galactomannan using guar gum, comprising the following steps: (1), washing guar gum sheets; (2), crushing; (3), enzymatically hydrolyzing; (4), ultrafiltering; (5), filtering with a decoloring membrane; 6), concentrating filtrate; (7), drying; (8), superfine grinding. The preparing method of the invention takes short time, saves production time and cost, and improves efficiency; matrix assisted laser desorption ionization-time of flight (MALDI-TOF) and high-performance liquid chromatography (HPLC) are used to determine the composition of differently-polymerized galactomannan in an enzymatic hydrolysate, enzymatic products meet the requirement for the polymerization degree of galactomannan, and monosaccharide component analysis determines that the ratio of mannose to galactose is 2:1.
Description
Technical field
The present invention relates to a kind of method utilizing guar gum quickly to prepare galactomannan, belong to biological technical field.
Background technology
Galactomannan (Galactomannan, write a Chinese character in simplified form GM) or claim galactomannan, it it is a kind of polysaccharide containing the key base other with galactose of mannose, more accurately for 1, galactomannan is the key polysaccharide being connected to α-D type galactose (alpha-D-galactose) in they 6-junction points of β-D type mannose ((1-4)-linkedbeta-D-mannopyranose) of linearity (1-4)-link, i.e. the α-D type galactopyranose (1-6-linkedalpha-D-galactopyranose) that 1-6-links.Activities of Some Plants and fungus all contain the composition of galactomannan.
According to the ratio of mannose Yu galactose, galactomannan kind is arranged from small to large by ratio:
Combination of fenugreek gum, mannose is than galactose about to；Melon glue (guargum), mannose is more two-to-one than galactose；Tara gum (taragum), mannose is than galactose about three to；Locust bean gum (locustbeangum, LBG) or carob (carobgum), mannose is than galactose about four to.Galactomannan is often employed in food product, is used for increasing the viscosity of its aqueous phase, it is also possible to the tranquilizer as food uses.It addition, galactomannan is also used among medicine and beauty treatment.Similar white powder, odorless, tasteless, acidproof, salt tolerant, Heat stability is good.Water soluble, aqueous solution is transparent, in neutral and have the lowest viscosity.More than LD505g/kg.This product has the lowest heat, has different physiological roles, can promote that little enteral bacillus bifidus is bred, Constipation, colon cancer, cardiovascular diseases and blood sugar lowering.Appropriate use can be needed by producing in varieties of food items.
Summary of the invention
The technical problem to be solved is: provide a kind of method utilizing guar gum quickly to prepare galactomannan.
For solving above-mentioned technical problem, the technical solution used in the present invention is:
What the present invention provided utilizes the method that guar gum quickly prepares galactomannan, comprises the following steps:
(1) guar splits is cleaned: guar splits cleaned 2-3 time with clear water, obtains clean sheet, then dries to moisture 15%；
(2) broken: the guar splits of drying to be carried out break process, it is thus achieved that the powdery of 10 mesh；
(3) enzymolysis: the guar gum of step (2) 100 parts by weight is dissolved in the warm water of 30-50 DEG C, add stearic acid monoglycerides and the citric acid of 15-20 parts by weight of 10-15 parts by weight, stirring warm water is until guar gum is completely dissolved, and guar gum mass concentration is at 20%-30%；Then it is warmed up to 60-75 DEG C, adds the organic acid of 5-10 weight portion, maintain 1.5-2h；It is rapidly cooled to 30 DEG C, adjusts pH to neutral with 5mol/LNaOH, add compound beta-mannase enzymolysis, after adding enzyme, temperature is warmed up to 45-55 DEG C, keep 3-4h, high temperature enzyme denaturing；
Add mixed enzyme and continue enzymolysis, temperature 40-50 DEG C, keep 2-3h, high temperature enzyme denaturing；
(4) ultrafiltration: use ultrafiltration apparatus, application specification 50kD ultrafilter membrane, be 30-40 DEG C in temperature, operation pressure is to carry out ultrafiltration under the conditions of 0.10-0.40MPa；
(5) decolouring membrane filtration: the solution after step (4) ultrafiltration is crossed decolouring film, and decolouring film is polyacrylamide film, and retaining molecular weight is 600-800Da, and operation temperature is 20-30 DEG C, and operation pressure is 0.20-0.30MPa；
(6) filtrate concentrates: taking the solution after decolouring membrane filtration and concentrate, thickening temperature 70-80 DEG C, after making concentration, the mass concentration of the solids in concentrated solution controls at 60%-70%；
(7) it is dried: using cryogenic vacuum continuous drying, absolute pressure is 0.01-0.1MPa, and temperature is 0-5 DEG C, is dried 7-9 hour；
(8) micronizing, is ground into the granule of 200 mesh, then sieves packaging.
Preferably, the organic acid mixture of one or more in citric acid, malic acid, tartaric acid, oxalic acid in step (3).Further, organic acid is selected from malic acid and tartaric mixture, and both part by weight are 3-5:1.
Preferably, step (3) compound beta-mannase includes α-galactomannan enzyme and xylanase, adds unit and is: the guar gum of every 100 parts by weight adds α-galactomannan enzyme 500-1000U, xylanase 400～600U.
Preferably, step (3) mixed enzyme includes pectase, cathepsin and papain, adds unit and is: the guar gum of every 100 parts by weight adds pectase 300～500U, cathepsin 500-1000U, papain 400～600U.
The preparation method of galactomannan mainly uses enzymatic isolation method at present, and the present invention combines acidolysis and enzymolysis, it is achieved quickly prepare the purpose of galactomannan, and step (1) carries out acidolysis by adding organic acid, the carrying out of the stearic acid monoglycerides of addition beneficially acidolysis.Step (2) carries out enzymolysis, by improving combination and the method for enzymolysis, it is thus achieved that optimal preparation effect.
The invention has the beneficial effects as follows:
1, enzymolysis while the present invention realizes α-galactomannan enzyme, pectase and xylanase, the time is short, eliminates the purification step after every kind of enzyme is degraded successively, has saved production time and cost, improve efficiency.
2, measuring the composition of different polymerization degree galactomannan in enzymolysis solution by Matrix-assisted laser desorption ionization (MALDI-TOF) and high performance liquid chromatography (HPLC), enzymatic hydrolysate meets galactomannan degree of polymerization requirement.Through monosaccharide composition analysis, the ratio recording wherein mannose and galactose is 2:1.
Detailed description of the invention
Embodiment 1
What the present embodiment provided utilizes the method that guar gum quickly prepares galactomannan, comprises the following steps:
(1) guar splits is cleaned: guar splits cleaned 3 times with clear water, obtains clean sheet, then dries to moisture 15%；
(2) broken: the guar splits of drying to be carried out break process, it is thus achieved that the powdery of 10 mesh；
(3) enzymolysis: the guar gum of step (2) 100 parts by weight is dissolved in the warm water of 50 DEG C, adds stearic acid monoglycerides and the citric acid of 20 parts by weight of 15 parts by weight, and stirring warm water is until guar gum is completely dissolved, and guar gum mass concentration is 20%；Then it is warmed up to 75 DEG C, adds the organic acid of 10 weight portions, maintain 2h；It is rapidly cooled to 30 DEG C, adjusts pH to neutral with 5mol/LNaOH, add compound beta-mannase enzymolysis, after adding enzyme, temperature is warmed up to 55 DEG C, keep 4h, high temperature enzyme denaturing；Add mixed enzyme and continue enzymolysis, temperature 40-50 DEG C, keep 2-3h, high temperature enzyme denaturing；
(4) ultrafiltration: using ultrafiltration apparatus, application specification 50kD ultrafilter membrane, is 40 DEG C in temperature, operation pressure is to carry out ultrafiltration under the conditions of 0.40MPa；
(5) decolouring membrane filtration: the solution after step (4) ultrafiltration is crossed decolouring film, and decolouring film is polyacrylamide film, and retaining molecular weight is 800Da, and operation temperature is 30 DEG C, and operation pressure is 0.30MPa；
(6) filtrate concentrates: taking the solution after decolouring membrane filtration and concentrate, thickening temperature 70 DEG C, after making concentration, the mass concentration of the solids in concentrated solution controls 60%；
(7) it is dried: using cryogenic vacuum continuous drying, absolute pressure is 0.1MPa, and temperature is 5 DEG C, is dried 9 hours；
(8) micronizing, is ground into the granule of 200 mesh, then sieves packaging.
In step (3), organic acid is selected from citric acid, the mixture of oxalic acid, and both part by weight are 5:1.
Step (3) compound beta-mannase includes α-galactomannan enzyme and xylanase, adds unit and is: the guar gum of every 100 parts by weight adds α-galactomannan enzyme 1000U, xylanase 600U.
Step (3) mixed enzyme includes pectase, cathepsin and papain, adds unit and is: the guar gum of every 100 parts by weight adds pectase 500U, cathepsin 1000U, papain 600U.
Embodiment
2
What the present embodiment provided utilizes the method that guar gum quickly prepares galactomannan, comprises the following steps:
(1) guar splits is cleaned: guar splits cleaned 2 times with clear water, obtains clean sheet, then dries to moisture 15%；
(2) broken: the guar splits of drying to be carried out break process, it is thus achieved that the powdery of 10 mesh；
(3) enzymolysis: the guar gum of step (2) 100 parts by weight is dissolved in the warm water of 30 DEG C, adds stearic acid monoglycerides and the citric acid of 15 parts by weight of 10 parts by weight, and stirring warm water is until guar gum is completely dissolved, and guar gum mass concentration is 30%；Then it is warmed up to 60 DEG C, adds the organic acid of 5 weight portions, maintain 1.5h；It is rapidly cooled to 30 DEG C, adjusts pH to neutral with 5mol/LNaOH, add compound beta-mannase enzymolysis, after adding enzyme, temperature is warmed up to 45 DEG C, keep 3h, high temperature enzyme denaturing；Add mixed enzyme and continue enzymolysis, temperature 40 DEG C, keep 2h, high temperature enzyme denaturing；
(4) ultrafiltration: using ultrafiltration apparatus, application specification 50kD ultrafilter membrane, is 30 DEG C in temperature, operation pressure is to carry out ultrafiltration under the conditions of 0.10MPa；
(5) decolouring membrane filtration: the solution after step (4) ultrafiltration is crossed decolouring film, and decolouring film is polyacrylamide film, and retaining molecular weight is 600Da, and operation temperature is 20 DEG C, and operation pressure is 0.20MPa；
(6) filtrate concentrates: taking the solution after decolouring membrane filtration and concentrate, thickening temperature 80 DEG C, after making concentration, the mass concentration of the solids in concentrated solution controls 70%；
(7) it is dried: using cryogenic vacuum continuous drying, absolute pressure is 0.01MPa, and temperature is 0 DEG C, is dried 7 hours；
(8) micronizing, is ground into the granule of 200 mesh, then sieves packaging.
In step (3), organic acid is selected from citric acid, malic acid, tartaric mixture.The part by weight of three is 3:5:1.
Step (3) compound beta-mannase includes α-galactomannan enzyme and xylanase, adds unit and is: the guar gum of every 100 parts by weight adds α-galactomannan enzyme 500U, xylanase 400U.
Step (3) mixed enzyme includes pectase, cathepsin and papain, adds unit and is: the guar gum of every 100 parts by weight adds pectase 300U, cathepsin 500U, papain 400U.
Embodiment
3
What the present embodiment provided utilizes the method that guar gum quickly prepares galactomannan, comprises the following steps:
(1) guar splits is cleaned: guar splits cleaned 3 times with clear water, obtains clean sheet, then dries to moisture 15%；
(2) broken: the guar splits of drying to be carried out break process, it is thus achieved that the powdery of 10 mesh；
(3) enzymolysis: the guar gum of step (2) 100 parts by weight is dissolved in the warm water of 40 DEG C, adds stearic acid monoglycerides and the citric acid of 18 parts by weight of 13 parts by weight, and stirring warm water is until guar gum is completely dissolved, and guar gum mass concentration is 25%；Then it is warmed up to 68 DEG C, adds the organic acid of 8 weight portions, maintain 2h；It is rapidly cooled to 30 DEG C, adjusts pH to neutral with 5mol/LNaOH, add compound beta-mannase enzymolysis, after adding enzyme, temperature is warmed up to 51 DEG C, keep 3.5h, high temperature enzyme denaturing；Add mixed enzyme and continue enzymolysis, temperature 45 C, keep 2.5h, high temperature enzyme denaturing；
(4) ultrafiltration: using ultrafiltration apparatus, application specification 50kD ultrafilter membrane, is 36 DEG C in temperature, operation pressure is to carry out ultrafiltration under the conditions of 0.30MPa；
(5) decolouring membrane filtration: the solution after step (4) ultrafiltration is crossed decolouring film, and decolouring film is polyacrylamide film, and retaining molecular weight is 700Da, and operation temperature is 26 DEG C, and operation pressure is 0.25MPa；
(6) filtrate concentrates: taking the solution after decolouring membrane filtration and concentrate, thickening temperature 75 DEG C, after making concentration, the mass concentration of the solids in concentrated solution controls 65%；
(7) it is dried: using cryogenic vacuum continuous drying, absolute pressure is 0.06MPa, and temperature is 3 DEG C, is dried 8 hours；
(8) micronizing, is ground into the granule of 200 mesh, then sieves packaging.
In step (3), organic acid is selected from malic acid and tartaric mixture, and both part by weight are 3-5:1.
Step (3) compound beta-mannase includes α-galactomannan enzyme and xylanase, adds unit and is: the guar gum of every 100 parts by weight adds α-galactomannan enzyme 8000U, xylanase 500U.
Step (3) mixed enzyme includes pectase, cathepsin and papain, adds unit and is: the guar gum of every 100 parts by weight adds pectase 400U, cathepsin 6000U, papain 500U.
The galactomannan preparing above-described embodiment is analyzed:
(1) organoleptic requirements
(2) thin-layer chromatographic analysis
Solution is prepared
Solution to be measured: be contained in Thick Centrifugal pipe by galactomannan prepared by the method for the 10mg present invention, adds the trifluoroacetic acid solution wiring solution-forming of 2mL 230 g/L, acutely rocks and make established colloid dissolve, and is closed the lid by centrifuge tube and heats 1h in 120 DEG C.Centrifugal, supernatant is transferred in the flask of 50 mL, adds 10 mL water, vacuum drying.Being dissolved in 10mL water by dried residue, be again vacuum dried, obtain the film of the clear without acetic acid odor, add 0.1 mL water and 1 mL methanol, centrifugal segregation precipitates, with methanol dilution supernatant to 1mL.
Standard solution: 10 mg galactose and 10 mg mannose being dissolved in 2 mL water, again with methanol is diluted to 10 mL.
Chromatographic condition and analytical procedure
Chromatoplate: tlc silica gel plate
Flowing phase: water: acetonitrile (volume ratio is 15:85)
Sample-adding amount: 5 μ L, puts into the band of 20 mm × 3 mm.
Launch: run sample more than 15 cm.
Spray with aminohippuric Acid, and heat 5 min at 120 DEG C.
Result
Standard solution chromatogram has 2 brown stripes being clearly separated, respectively galactose and mannose；The chromatogram of solution to be measured also showing that, 2 band, corresponding position are respectively galactose and mannose.
Claims (5)
1. one kind utilizes the method that galactomannan quickly prepared by guar gum, it is characterised in that comprise the following steps:
(1) guar splits is cleaned: guar splits cleaned 2-3 time with clear water, obtains clean sheet, then dries to moisture 15%；
(2) broken: the guar splits of drying to be carried out break process, it is thus achieved that the powdery of 10 mesh；
(3) enzymolysis: the guar gum of step (2) 100 parts by weight is dissolved in the warm water of 30-50 DEG C, add stearic acid monoglycerides and the citric acid of 15-20 parts by weight of 10-15 parts by weight, stirring warm water is until guar gum is completely dissolved, and guar gum mass concentration is at 20%-30%；Then it is warmed up to 60-75 DEG C, adds the organic acid of 5-10 weight portion, maintain 1.5-2h；It is rapidly cooled to 30 DEG C, adjusts pH to neutral with 5mol/LNaOH, add compound beta-mannase enzymolysis, after adding enzyme, temperature is warmed up to 45-55 DEG C, keep 3-4h, high temperature enzyme denaturing；
Add mixed enzyme and continue enzymolysis, temperature 40-50 DEG C, keep 2-3h, high temperature enzyme denaturing；
(4) ultrafiltration: use ultrafiltration apparatus, application specification 50kD ultrafilter membrane, be 30-40 DEG C in temperature, operation pressure is to carry out ultrafiltration under the conditions of 0.10-0.40MPa；
(5) decolouring membrane filtration: the solution after step (4) ultrafiltration is crossed decolouring film, and decolouring film is polyacrylamide film, and retaining molecular weight is 600-800Da, and operation temperature is 20-30 DEG C, and operation pressure is 0.20-0.30MPa；
(6) filtrate concentrates: taking the solution after decolouring membrane filtration and concentrate, thickening temperature 70-80 DEG C, after making concentration, the mass concentration of the solids in concentrated solution controls at 60%-70%；
(7) it is dried: using cryogenic vacuum continuous drying, absolute pressure is 0.01-0.1MPa, and temperature is 0-5 DEG C, and moisture Control is at 10-12%；
(8) micronizing, is ground into the powdery of 200 mesh, and moisture Control is less than 5%, then sieves packaging.
The method utilizing guar gum quickly to prepare galactomannan the most according to claim 1, it is characterised in that the organic acid mixture of one or more in citric acid, malic acid, tartaric acid, oxalic acid in step (3).
The method utilizing guar gum quickly to prepare galactomannan the most according to claim 2, it is characterised in that organic acid is selected from malic acid and tartaric mixture, and both part by weight are 3-5:1.
The method utilizing guar gum quickly to prepare galactomannan the most according to claim 1, it is characterized in that, step (3) compound beta-mannase includes α-galactomannan enzyme and xylanase, adding unit is: the guar gum of every 100 parts by weight adds α-galactomannan enzyme 500-1000U, xylanase 400～600U.
The method utilizing guar gum quickly to prepare galactomannan the most according to claim 1, it is characterized in that, step (3) mixed enzyme includes pectase, cathepsin and papain, adding unit is: the guar gum of every 100 parts by weight adds pectase 300～500U, cathepsin 500-1000U, papain 400～600U.
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Cited By (9)
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| CN106418554A (en) * | 2016-09-07 | 2017-02-22 | 中国农业大学 | Complex enzyme hydrolysis guar gum preparation soluble diet fiber and manna oligose method |
| CN106755194A (en) * | 2016-12-12 | 2017-05-31 | 青州荣美尔生物科技股份有限公司 | A kind of preparation technology of ultra-low viscosity guar gum |
| CN108917292A (en) * | 2018-05-16 | 2018-11-30 | 湖南长沙天地人生物科技有限公司 | A kind of drug sensitive test card drying means |
| CN110438190A (en) * | 2019-09-12 | 2019-11-12 | 北京瓜尔润科技股份有限公司 | A kind of method and Guar active peptide using a variety of enzyme complex enzyme hydrolysis preparation Guar active peptide |
| CN111685333A (en) * | 2020-06-15 | 2020-09-22 | 北京瓜尔润科技股份有限公司 | Extraction method of water-soluble dietary fiber |
| CN111690701A (en) * | 2020-06-15 | 2020-09-22 | 北京瓜尔润科技股份有限公司 | Separation and purification method of galactomannan |
| CN113575955A (en) * | 2021-07-02 | 2021-11-02 | 青岛凯特生物科技有限公司 | Prebiotic composition and preparation method thereof |
| CN114403228A (en) * | 2022-01-19 | 2022-04-29 | 杭州威欧生物科技有限公司 | Stabilizer for acidic milk-containing beverage and application thereof |
| CN114908197A (en) * | 2022-06-28 | 2022-08-16 | 安徽禾庚生物技术有限公司 | Method for producing galactose and mannose by taking guar gum as raw material |
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| CN106418554A (en) * | 2016-09-07 | 2017-02-22 | 中国农业大学 | Complex enzyme hydrolysis guar gum preparation soluble diet fiber and manna oligose method |
| CN106418554B (en) * | 2016-09-07 | 2019-12-20 | 中国农业大学 | Method for preparing soluble dietary fiber and mannan oligosaccharide by hydrolyzing guar gum with complex enzyme |
| CN106755194A (en) * | 2016-12-12 | 2017-05-31 | 青州荣美尔生物科技股份有限公司 | A kind of preparation technology of ultra-low viscosity guar gum |
| CN108917292A (en) * | 2018-05-16 | 2018-11-30 | 湖南长沙天地人生物科技有限公司 | A kind of drug sensitive test card drying means |
| CN110438190A (en) * | 2019-09-12 | 2019-11-12 | 北京瓜尔润科技股份有限公司 | A kind of method and Guar active peptide using a variety of enzyme complex enzyme hydrolysis preparation Guar active peptide |
| CN110438190B (en) * | 2019-09-12 | 2021-07-30 | 北京瓜尔润科技股份有限公司 | Method for preparing guar active peptide by using multiple enzyme composite enzymolysis and guar active peptide |
| CN111685333A (en) * | 2020-06-15 | 2020-09-22 | 北京瓜尔润科技股份有限公司 | Extraction method of water-soluble dietary fiber |
| CN111690701A (en) * | 2020-06-15 | 2020-09-22 | 北京瓜尔润科技股份有限公司 | Separation and purification method of galactomannan |
| CN113575955A (en) * | 2021-07-02 | 2021-11-02 | 青岛凯特生物科技有限公司 | Prebiotic composition and preparation method thereof |
| CN114403228A (en) * | 2022-01-19 | 2022-04-29 | 杭州威欧生物科技有限公司 | Stabilizer for acidic milk-containing beverage and application thereof |
| CN114908197A (en) * | 2022-06-28 | 2022-08-16 | 安徽禾庚生物技术有限公司 | Method for producing galactose and mannose by taking guar gum as raw material |
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