CN109609572A - A kind of preparation method of the mannosan of specified molecular weight - Google Patents
A kind of preparation method of the mannosan of specified molecular weight Download PDFInfo
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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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Abstract
The invention discloses a kind of methods that hemicellulose enzymolysis prepares the mannosan of specified molecular weight, main to use a kind of special enzymatic hydrolysis system progress Controlled-enzymatic Hydrolysis, which includes enzymatic hydrolysis container, twin-screw propulsion device, digital display temperature-control system, molecular weight on-line checking digital display device, gear pump.With the enzymatic hydrolysis system can accurately controlled enzymatic hydrolysis degree, obtain the mannosan of required molecular weight or molecular weight segment；The problem of it is high that the enzymatic hydrolysis system solves hemicellulose enzymolysis process medium viscosity, and while feeding intake easily agglomerates, improves enzymatic hydrolysis rate, and under equal conditions, compared with unmodified enzymolysis device, enzymolysis process shortens one third to the time of half；Meanwhile enzymatic hydrolysis and enzyme deactivation can carry out in same container, are not necessarily to external enzyme-removal equipment, simplify production procedure, Synergistic and energy-saving.
Description
Technical field
The invention belongs to biological chemical fields, and in particular to a kind of preparation facilities of the mannosan of specified molecular weight and side
Method.
Background technique
Dew glycan is with the linear polysaccharide of β-Isosorbide-5-Nitrae-D- mannopyranoside bonds connection, if the certain residues of main chain are by Portugal
Grape sugar replaces or galactolipin passes through 1,6- α-glycosidic bond and mannose residue is connected to form branch, then referred to as different mannosan,
Mainly there are galactomannans, glucomannans and galactoglucomannan.Due to constituting monosaccharide type, the polymerization of these glycan
Degree, sugar key connection and arrangement mode etc. are different, and resulting in different glycan, there is apparent in nature in solubility, viscosity etc.
Difference greatly affected the hydrolysis of 'beta '-mannase.Therefore the application of mannosan is limited to a certain extent.
In the research of mannosan enzymatic degradation, most of research concentrates on that mannosan is degraded into molecular weight very
Low oligosaccharide, Oligomeric manna sugar are the lower a kind of sugar of the degree of polymerization, and the degree of polymerization of glycan molecule is in 2-10 sugar, extensively
Applied to food, chemical industry and medicine and other fields.The research of mannosan about other molecular weight sections is less, is mainly related to sweet
Dew big sugared (patent 201711340163.4), sweet dew time glycan (201711340162.X), the sub- glycan of sweet dew time
(201711341567.5) preparation patent, the mannosan of these molecular weight segments have different functional characteristics, respectively
It applies in different fields, but the molecular weight ranges of mannosan (ten thousand Da of 400Da-120) are relatively wider, there are also a lot of other spies
Determine the excavation to be prepared with functional characteristic such as mannosan of molecular weight segment.
Researcher prepares the mannosan of different molecular weight at present, and production is controlled generally by enzymolysis time and enzyme concentration
The molecular weight of object, randomness is big, and reproducibility is not high, and the molecular weight of the product of preparation is not also comprehensive.Some are by controlling molecule
Amount size carrys out controlled enzymatic hydrolysis, such as auxiliary using gel permeation chromatography-laser light scattering joint technology, nuclear magnetic resonance spectroscopy, matrix
The technologies such as laser desorption/ionization time of flight mass mass spectrum are helped, the molecular weight of enzymolysis liquid can be measured, measurement result accuracy is good,
But it cannot achieve real-time online detection since detection program is complicated, time-consuming.And viscosimetry is despite a kind of molecular weight
Indirect Detecting Method, but instrument and equipment is simple, operates conveniently, and the molecular weight scope of application is big, and has similar detection accuracy, can
Go out viscosity with on-line real-time measuremen, molecular size range is obtained by the relational expression of viscosity and molecular weight.
The invention patent will solve the problems, such as it is that molecular weight on-line measuring device is arranged on enzymatic hydrolysis container, by examining online
The size of molecular weight is surveyed, accurately controlled enzymatic hydrolysis degree, obtains the mannosan of any required molecular weight or molecular weight segment.
When enzymatic hydrolysis prepares the mannosan of different molecular weight segment, since feed particles are relatively fine, very when feeding intake
It is easy agglomeration, is especially fed intake when carrying out half, enzymolysis liquid is very sticky, and when continuing spreading, some tiny material particles can quilts
It is wrapped in the inside, and cannot sufficiently be digested.It is avoided when feeding intake in the prior art for konjaku, guar gum, locust bean gum etc.
The solution of agglomeration problems, being mainly concerned with reduces concentration of substrate, stirring, addition ion or is located in advance with certain reagents
Reason.It by reducing concentration of substrate, avoidable agglomeration, but is used in large-scale production, efficiency is very low；Stirring is applied to scale
Low efficiency in production, and caking phenomenon can still occur because stirring is uneven；Add ion or at certain reagents
Reason, is easy to remain in catabolite, increases the difficulty of purifying, limit its application in food and field of medicaments.
The present invention carries out Controlled-enzymatic Hydrolysis using a kind of special enzymatic hydrolysis system, which includes enzymatic hydrolysis container, double spiral shells
Bar propulsion device, digital display temperature-control system, molecular weight on-line checking digital display device, gear pump can solve high viscosity and the knot that feeds intake
Block problem improves enzymatic hydrolysis rate；Meanwhile enzymatic hydrolysis and enzyme deactivation can carry out in same enzymatic hydrolysis container, be not necessarily to external enzyme-removal equipment,
Simplify production procedure, Synergistic and energy-saving.
Summary of the invention
It is main to use a kind of special enzyme the invention discloses a kind of production method of the mannosan of specified molecular weight
Solution system carries out Controlled-enzymatic Hydrolysis, which includes enzymatic hydrolysis container, twin-screw propulsion device, digital display temperature-control system, molecular weight
On-line checking digital display device, gear pump.With the enzymatic hydrolysis system can accurately controlled enzymatic hydrolysis degree, obtain required molecular weight or
The mannosan of molecular weight segment.
Under the promotion of twin-screw propulsion device and gear pump, enzymatic hydrolysis circulation carries out feed liquid, can solve hemicellulase
Solution preocess medium viscosity is high, and while feeding intake easy agglomeration problems, improves enzymatic hydrolysis rate, under equal conditions, compared with original, digests
Journey shortens one third to the time of half；In the lower outlet port of enzymatic hydrolysis container, it is provided with molecular weight on-line checking number
Aobvious device, when detecting target molecular weight size, digital display temperature-control system can start twin-screw automatically according to the set procedure and push away
Heating function into device heats enzyme deactivation；The temperature range of the digital display temperature-control system is at 0-150 DEG C.In substrate feed liquid enzymatic hydrolysis,
Temperature auto-control is 30-35 DEG C, when enzyme deactivation, and temperature auto-control is 100-105 DEG C, and enzymatic hydrolysis and enzyme deactivation can be in same containers
It carries out, is not necessarily to external enzyme-removal equipment, simplify production procedure, Synergistic and energy-saving.
To obtain above-mentioned technical effect, the technical scheme adopted by the invention is that:
Hemicellulose using konjac glucomannan, guar gum, locust bean gum, sesbania gum etc. containing mannosan uses a kind of spy as raw material
Different enzymatic hydrolysis system carries out Controlled-enzymatic Hydrolysis, and passes through the series of processes such as centrifugation, UF membrane, ion exchange, needed for finally obtaining
The mannosan of molecular weight or molecular weight segment.
The preparation process of the mannosan of specified molecular weight is as follows:
Hemicellulose → Controlled-enzymatic Hydrolysis → centrifuge separation → UF membrane → ion exchange → spray drying → acquisition specified molecular weight
Mannosan
During preparing the mannosan of specified molecular weight, this procedure of Controlled-enzymatic Hydrolysis is the most key, and the present invention is to existing
Enzymatic vessel and enzyme deactivation device carried out creative integrated design, establish a kind of special enzymatic hydrolysis system, including enzymatic hydrolysis
Container 1, twin-screw propulsion device 2, digital display temperature-control system 3, molecular weight on-line checking digital display device 4, gear pump 5；
The temperature range of digital display temperature-control system is at 0-150 DEG C in the enzymatic hydrolysis system, and in substrate feed liquid enzymatic hydrolysis, temperature is adjusted automatically
Control is 30-35 DEG C, when enzyme deactivation, and temperature auto-control is 100-105 DEG C；
The enzymatic hydrolysis system middle-molecular-weihydroxyethyl on-line checking digital display device 4 is made of sensor probe and transmitter, sensor vibration
Dynamic frequency probe is 50-80Hz, amplitude 1-2mm；Principle design is shaken according to torque, uniform amplitude is stablized by electric current.Work
When making, after probe sensor is completely immersed in enzymolysis liquid, since the enzymolysis liquid of different Degree of Enzymatic Hydrolysis has different viscosity, make
It obtains between enzymolysis liquid and transducer probe face, generates the phase change of various amplitude, and sensor probe is to protect always at this time
Identical micro- amplitude resonance shearing wave is held, the variation of fluid viscosity is believed by electric current of the maintenance sensor probe under fixed amplitude
Number reflection, the current signal that this change by transmitter is converted into the viscosity number of enzymolysis liquid, then according to viscosity number in program with divide
The relational expression of son amount, is converted to molecular weight values.
Specifically in the on-line checking of the enzymolysis process middle-molecular-weihydroxyethyl of mannosan, with the progress that mannosan digests, enzyme
The reduction of fluid viscosity is solved, maintains electric current of the sensor probe under fixed amplitude that can also reduce therewith, and then convert by program
For the molecular size range of mannosan, and shown on digital display device.
Setting is used for the feeding port 6 of adding raw materials and enzyme on the enzymatic hydrolysis container 1, for releasing the discharge port 7 of enzymolysis liquid；
In further technical solution, the mannosan of the specified molecular weight of acquisition can be unimodal molecular weight component or a certain
Molecular weight segment component.
Compared with prior art, the integrated design of enzymatic hydrolysis of the invention and enzyme deactivation process, has the advantages that
(1) required enzymatic hydrolysis node is determined by molecular weight on-line checking digital display device, when detecting target molecular weight size
When, the heating function that digital display temperature-control system can start automatically twin-screw propulsion device according to setting program quickly heats, accomplish and
When enzyme deactivation, the enzymolysis product of required molecular weight ranges accurately can be easily obtained in industrialized production.
(2) enzymatic hydrolysis system is equipped with twin-screw propulsion device and gear pump, and feed liquid is under their common promotion, enzymatic hydrolysis circulation
It carries out, easy agglomeration problems when it is high to can solve hemicellulose enzymolysis process medium viscosity, and feeding intake improve enzymatic hydrolysis rate, enzymatic hydrolysis
Process shortens one third to the time of half.
(3) enzymatic hydrolysis and enzyme deactivation carry out in same tankage, are not necessarily to external enzyme-removal equipment, simplify production procedure, increase
Effect energy conservation.
Detailed description of the invention
Fig. 1 is special enzymatic hydrolysis system and device of the invention.
Specific embodiment
Embodiment 1
The industrial water that 450kg is injected in container will be digested, temperature is 30 DEG C, pH 6.0-7.0 in adjustment enzymatic hydrolysis container, to
It digests and puts into neutral β-Mannannase (input amount of enzyme is 1:15 calculating according to enzyme material mass ratio) in container, then weigh
50kg konjaku powder is launched into enzymatic hydrolysis container, and under the promotion of twin-screw propulsion device and gear pump, enzymatic hydrolysis circulation is carried out,
The vibration probe frequency for adjusting molecular weight on-line checking digital display device is 80Hz, amplitude 1mm, with the progress of enzymatic hydrolysis, enzymatic hydrolysis
Liquid viscosity is gradually reduced, when molecular weight on-line checking digital display device detects that the molecular weight of enzymolysis liquid is 100000-200000Da
When, digital display temperature-control system starts the heating function of twin-screw propulsion device automatically according to the set procedure, makes to digest temperature in container
Degree is rapidly achieved 100 DEG C of progress enzyme deactivations, and the enzyme deactivation time is 20-30min, cooling；Then pass through centrifugation, UF membrane, ion exchange
Etc. series of processes, finally obtain molecular weight in the mannosan of 100000-200000Da.
Embodiment 2
The industrial water that 450kg is injected in container will be digested, temperature is 30 DEG C, pH 6.0-7.0 in adjustment enzymatic hydrolysis container, to
It digests and puts into neutral β-Mannannase (input amount of enzyme is 1:15 calculating according to enzyme material mass ratio) in container, then weigh
50kg guar gum is launched into enzymatic hydrolysis container, under the promotion of twin-screw propulsion device and gear pump, carries out circulation enzymatic hydrolysis,
The vibration probe frequency for adjusting molecular weight on-line checking digital display device is 80Hz, amplitude 1mm, with the progress of enzymatic hydrolysis, enzymatic hydrolysis
Liquid viscosity is gradually reduced, when molecular weight on-line checking digital display device detects that the molecular weight of enzymolysis liquid is 10000-12000Da,
Digital display temperature-control system starts the heating function of twin-screw propulsion device automatically according to the set procedure, and it is fast to make to digest temperature in container
Speed reaches 100 DEG C of progress enzyme deactivations, and the enzyme deactivation time is 20-30min, cooling；Then pass through centrifugation, UF membrane, ion exchange etc. one
Serial process finally obtains molecular weight in the mannosan of 10000-12000Da.
Embodiment 3
The industrial water that 450kg is injected in container will be digested, temperature is 30 DEG C, pH 6.0-7.0 in adjustment enzymatic hydrolysis container, to
It digests and puts into neutral β-Mannannase (input amount of enzyme is 1:15 calculating according to enzyme material mass ratio) in container, then weigh
50kg locust bean gum is launched into enzymatic hydrolysis container, under the promotion of twin-screw propulsion device and gear pump, carries out circulation enzymatic hydrolysis, adjusts
The vibration probe frequency of whole molecule amount on-line checking digital display device is 50Hz, amplitude 2mm, with the progress of enzymatic hydrolysis, enzymolysis liquid
Viscosity is gradually reduced, when molecular weight on-line checking digital display device detects that the molecular weight of enzymolysis liquid is 3000-5000Da, digital display
Temperature control system starts the heating function of twin-screw propulsion device automatically according to the set procedure, makes to digest temperature in container and quickly reaches
To 105 DEG C of progress enzyme deactivations, the enzyme deactivation time is 20-30min, cooling；It is a series of to then pass through centrifugation, UF membrane, ion exchange etc.
Process finally obtains molecular weight in the mannosan of 3000-5000Da.
Above description is not limitation of the present invention, and the present invention is also not limited to the example above.The art it is common
Within the essential scope of the present invention, the variations, modifications, additions or substitutions made also should belong to protection of the invention to technical staff
Range.
Claims (3)
1. a kind of mannosan preparation method of specified molecular weight, which is characterized in that with konjac glucomannan, guar gum, locust bean gum, field
The hemicellulose containing mannosan such as cyanines glue is raw material, carries out Controlled-enzymatic Hydrolysis, zymolyte using a kind of special enzymatic hydrolysis system
By isolating and purifying to obtain the mannosan of required molecular weight or molecular weight segment.
2. as described in claim 1, a kind of special enzymatic hydrolysis system includes enzymatic hydrolysis container 1, twin-screw propulsion device 2, digital display temperature
Control system 3, molecular weight on-line checking digital display device 4, gear pump 5；
The temperature range of digital display temperature-control system 3 is at 0-150 DEG C in the enzymatic hydrolysis system, and in substrate feed liquid enzymatic hydrolysis, temperature is automatic
It is regulated to 30-35 DEG C, when enzyme deactivation, temperature auto-control is 100-105 DEG C；
The sensor vibration frequency probe of the enzymatic hydrolysis system middle-molecular-weihydroxyethyl on-line checking digital display device 4 is 50-80Hz, and amplitude is
1-2mm；
Setting is used for the feeding port 6 of adding raw materials and enzyme on the enzymatic hydrolysis container 1, for releasing the discharge port 7 of enzymolysis liquid.
3. as described in claim 1, the mannosan of specified molecular weight can be unimodal molecular weight component or a certain molecular weight
Segment component.
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| CN106755201A (en) * | 2017-03-23 | 2017-05-31 | 中国海洋大学 | A kind of preparation method of galactomannan oligosaccharide |
| CN107760740A (en) * | 2017-12-14 | 2018-03-06 | 宁波拜尔玛生物科技有限公司 | A kind of big sugar of high-purity kanjak mannan and preparation method thereof, application |
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2019
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Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN203613198U (en) * | 2013-11-18 | 2014-05-28 | 中国农业科学院农产品加工研究所 | Reaction system for preparing high molecular weight vegetable protein peptide |
| CN105567761A (en) * | 2014-10-16 | 2016-05-11 | 中粮营养健康研究院有限公司 | Enzymolysis method for cellulose-contained material |
| CN106755201A (en) * | 2017-03-23 | 2017-05-31 | 中国海洋大学 | A kind of preparation method of galactomannan oligosaccharide |
| CN207091430U (en) * | 2017-08-18 | 2018-03-13 | 成都长力元生物科技有限公司 | A kind of equipment of efficiently enzymolysis glucomannan |
| CN107760740A (en) * | 2017-12-14 | 2018-03-06 | 宁波拜尔玛生物科技有限公司 | A kind of big sugar of high-purity kanjak mannan and preparation method thereof, application |
| CN107982275A (en) * | 2017-12-14 | 2018-05-04 | 宁波拜尔玛生物科技有限公司 | Application of the big sugar of sweet dew in gynaecologic antibiotic product |
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| Title |
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| YU CHENG等: "Enzymatic Degradation of Guar and Substituted Guar Galactomannans", 《BIOMACROMOLECULES》 * |
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