CN104004800B - A kind of preparation method of amylose inclusion complexation carrier - Google Patents
A kind of preparation method of amylose inclusion complexation carrier Download PDFInfo
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Abstract
The invention discloses a kind of preparation method of amylose inclusion complexation carrier.The method starch material disodium hydrogen phosphate sodium dihydrogen phosphate buffer is made into starch milk, and adjusts pH to 4.5~6.5, is placed in closed container and is gelatinized；After the completion of gelatinization, dextrin is cooled down, add debranching enzyme, stirring reaction 1~3 hour at 45~65 DEG C；Regulation pH to 5.5~7.0, adds alpha amylase, and after 1~10 minute, go out stirring reaction enzyme at 45~110 DEG C；The iodine KI mixed solution that addition water is configured, it is 5~20mg/mL to control concentration of the KI in mixed solution；60 DEG C~80 DEG C insulated and stirreds 10~20 minutes；PH is neutralized to 6.0~6.5, be dry, pulverize.The materials such as amylose carrier prepared by the present invention, energy inclusion complexation essence and flavoring agent, aliphatic acid, inorganic molecules, have been widely used in industries such as food, medicine, cosmetics, chemical industry.
Description
Technical field
The present invention relates to the production method of converted starch, specifically refer to using biotechnology and the method system for adding derivant
For the method for the starch inclusion complexation carrier to function factor with loading functional.
Background technology
Starch is a kind of reproducible carbohydrate, is made up of two kinds of polysaccharide molecules, i.e. amylose and amylopectin.
Amylose is the straight-chain molecule being formed by connecting by α -1,4 glycosidic bonds；Amylopectin be then to be polymerized by D- glucopyranoses and
Into branch-like molecule, its linear fraction be α-Isosorbide-5-Nitrae glycosidic bond, and branch crunode then be α -1,6 glycosidic bonds.
Amylose in aqueous, typically in random Coiling-type, in the presence of having appropriate guest molecule in solution, directly
Chain starch molecule can form single-screw chain, the whole cylindrical cavity body structure of spiral.Structure to cyclodextrin is similar, amylose
The inner chamber of single-screw chain is hydrophobic and outside is hydrophilic, therefore amylose can be used as a kind of host molecule, by hydrophobic mutual
Act on and form Inclusion Complexes from different hydrophobic guest molecules.According to the of different sizes of guest molecule, each spiral can be by
6th, 7 or 8 glucose unit compositions, pitch can also change；Guest molecule can be located inside spiraled cavity, may be alternatively located at two
Between spiral.Amylose is to some inorganic matters (such as iodine, KBr, potassium hydroxide) and organic matter (such as dimethyl sulfoxide, alcohol, fat
Fat acid, surfactant, aromatic compound, dye molecule) inclusion complexation effect existing many reports in the literature.
Comprehensive research conditions both domestic and external, for study the amylose used by amylose inclusion complexation function be generally from
It is isolated in ative starch, it is relatively costly, it is unfavorable for industrialized production and the application of amylose inclusion complexation body.Using de- branch
Enzyme carries out de- branch to starch, can obtain substantial amounts of amylose molecule.Formed sediment by the controllable straight chain of AMS limited enzymatic hydrolysis
The molecular weight and the degree of polymerization of powder obtain being adapted for the amylose of inclusion complexation effect.Adding spiraled cavity derivant can promote
The formation of amylose single coil configuration, improves the inclusion complexation ability of amylose.It is domestic at present on using debranching enzyme and
AMS synergy prepares amylose and adds IKI derivant to prepare the side of amylose inclusion complexation carrier
Method yet there are no.
The content of the invention
It is an object of the invention to be directed to amylose preparation cost shortcoming high in the prior art, there is provided one kind is produced into
The preparation method of this low, efficiency high amylose inclusion complexation carrier.
A kind of preparation method of amylose inclusion complexation carrier of the present invention, comprises the following steps：
(1) starch material disodium hydrogen phosphate-sodium dihydrogen phosphate buffer be made into mass percent concentration for 5%~
12% starch milk, and pH to 4.5~6.5 is adjusted, it is placed in closed container and 30~60min of gelatinization is stirred at 95~99 DEG C；
(2) the dextrin cooling for obtaining step (1), adds debranching enzyme, and consumption is that every gram of dried starch adds debranching enzyme 10
~30u, stirring reaction 1~3 hour at 45~65 DEG C；
(3) pH to 5.5~7.0 is adjusted, AMS is added, consumption is that every gram of dried starch adds 1~10u of AMS,
Stirring reaction 1~10 minute at 45~110 DEG C；Regulation pH to 2.5~3.0 simultaneously keeps 10~15 minutes enzymes that go out；Addition water
The IKI mixed solution of configuration, controls concentration of the iodine in mixed solution for 0.5~2mg/mL, and control KI is mixed
It is 5~20mg/mL to close the concentration in solution；Consumption is that every gram of dried starch adds IKI mixed solution 0.5~2mL, 60~
80 DEG C of insulated and stirreds 10~20 minutes, obtain mixture；
(4) step (3) gained mixture is neutralized to pH6.0~6.5, amylose inclusion complexation is obtained after dry, pulverize
Carrier.
In order to preferably realize the present invention, step (1) described starch material be waxy corn starch, Waxy wheat starch,
One kind in tapioca, cornstarch, farina or wheaten starch；
Step (2) described debranching enzyme is Pullulanase or isoamylase；
Step (3) described AMS is middle temperature or Thermostable α-Amylase；
Drying described in step (4) is using spray drying or freeze-drying.
The present invention compared with prior art, has the following advantages that and effect：
(1) it is that amylose inclusion complexation carrier is prepared using the method for biological enzymolysis technology and addition IKI, drops
The low production cost of amylose, improves the inclusion complexation ability of amylose, is that the industrialization of amylose carrier should
With there is provided feasibility, and because preparation process does not need organic solvent, environmentally safe, energy-conserving and environment-protective.
(2) present invention has that production efficiency is high, high quality.The amylose inclusion complexation carrier for obtaining its
Up to 1.12, the amylose that will be obtained is used for inclusion complexation stearic acid to blue value, and stearic content is reachable in inclusion compound
11.2%, stearic acid embedding rate is up to 75%.With important Social benefit and economic benefit.
Brief description of the drawings
Fig. 1 is the UV, visible light of wheat ative starch/Surgidine and the gained wheat amylose of embodiment 1/Surgidine
Absorption spectrum.A is the ultraviolet-visible absorption spectroscopy of potato ative starch/Surgidine in figure；B is potato amylose/iodine
The ultraviolet-visible absorption spectroscopy of compound.
Specific embodiment
To be best understood from the present invention, the present invention is described further with reference to embodiment.The present invention have it is many into
The embodiment of work(, is set forth below six specific embodiments, but the scope of protection of present invention is not limited to embodiment
The scope of expression.
Embodiment 1
It is 5% that first step wheaten starch disodium hydrogen phosphate-sodium dihydrogen phosphate buffer is made into mass percent concentration
Starch milk, and adjust pH to 4.5, be placed in closed container in 95 DEG C of stirring gelatinization 30min；
The dextrin cooling that second step obtains step (1), (OPTIMAX L-1000, Jie Neng sections give birth to add Pullulanase
Thing Engineering Co., Ltd), consumption is that every gram of dried starch adds Pullulanase 10u, stirring reaction 3 hours at 60 DEG C；
3rd step section pH to 6.0, adds mesophilicα-diastase (CLARASE L, Jie Nengke bioengineering Co., Ltd),
Consumption is that every gram of dried starch adds mesophilicα-diastase 1u, and after 10 minutes, regulation pH to 2.5 simultaneously keeps stirring reaction at 50 DEG C
10 minutes enzymes that go out；The IKI mixed solution that addition is configured with water, it is 0.5mg/ to control concentration of the iodine in mixed solution
ML, it is 5mg/mL to control concentration of the KI in mixed solution；Consumption is that every gram of dried starch adds IKI mixed solution
2mL, 60 DEG C of insulated and stirreds 10 minutes, obtains mixture；
4th step neutralizes to pH6.0 step (3) gained mixture with 0.5mol/L hydrochloric acid, is spray-dried, after crushing
White powder amylose inclusion complexation carrier.After measured, the blue value of products obtained therefrom is 1.08.Use the amylose carrier
Stearic acid is embedded, the inclusion compound for obtaining shows after measured, stearic content is 10.1%, stearic acid bag in inclusion compound
It is 67.6% to bury rate.
Blue value is the important indicator for characterizing amylose inclusion complexation ability.Fig. 1 is for wheat ative starch/Surgidine and originally
The ultraviolet-visible absorption spectroscopy of embodiment gained wheat amylose/Surgidine.Ative starch is through debranching enzyme and AMS enzyme
Xie Hou, forms substantial amounts of amylose, as shown in figure 1, compound and former wheaten starch that wheat amylose is formed with iodine
Compared with the compound that iodine is formed, its absorption peak strength in the range of 550~700nm is remarkably reinforced, and shows wheat ative starch
Through digesting and after adding IKI derivant, its inclusion complexation ability is remarkably reinforced.
The specific assay method of blue value is：It is the sample 1mL of 0.5mg/mL to draw the good concentration of constant volume, and 1mol/L is added dropwise
NaOH solution 0.5mL, boiling water bath 3min, after cooling, add 1mol/L HCl solutions 0.5mL to neutralize, it is subsequently added 0.07~
0.1g potassium hydrogen tartrates, add water to nearly 45mL, are subsequently adding 0.5mL iodine solutions (2mg/mL I2,20mg/mL KI), are settled to
50mL volumetric flasks, mix, and after placing 20min at room temperature, absorbance are surveyed with 1cm cuvettes under 680nm, calculate blue with following formula
Value：
Blue value=(absorbance × 4)/sample concentration (mg/dL)
The specific method of stearic acid content measure is in inclusion compound：Precision weighs 1g composites of starch, is placed in 50mL test tubes
It is interior, add 5mL anhydrous hydrochloric acids after mixing again.Test tube is put into 70~80 DEG C of water-baths, one was stirred with glass bar every 5~10 minutes
It is secondary, untill treatments of the sample is complete, take out test tube within about 20~25 minutes, add the ethanol that 5mL concentration is 95%, mixing.It is cold
But after, by mixture immigration 50mL tool plug graduated cylinders, with 10mL absolute ethers, each 5mL cleans test tube at twice, pours into the lump
In graduated cylinder, after absolute ether all pours into graduated cylinder, plug is carefully opened in shaking 1 minute of jumping a queue, and releases gas, then is stoppered, and stands 12 points
Clock.It is careful to open plug, and the fat adhered to the equivalent mixed liquor washout plug and nozzle of 5mL dry oils ether and absolute ether.It is quiet
Put 10~20 minutes, treat that upper liquid is clear, suction out supernatant in the conical flask of constant weight, shake, after standing, will upper strata
Ether is suctioned out, and is put into former conical flask, conical flask is placed in water-bath and is evaporated, and is put in 105 DEG C of baking ovens and is dried 2 hours, and taking-up is put
Enter after being cooled down 0.5 hour in drier, weigh.The calculating of stearic acid content is as follows：
X=(m1-m0)/m2×100
In formula：X is stearic content, % in sample；m1It is conical flask and stearic quality, g；m0It is conical flask
Quality, g；m2It is the quality of sample, g.
The calculating of stearic embedding rate is as follows：
Stearic acid quality × 100 of stearic quality/input in embedding rate (%)=compound.
Embodiment 2
It is 12% that first step cornstarch disodium hydrogen phosphate-sodium dihydrogen phosphate buffer is made into mass percent concentration
Starch milk, and adjust pH to 6.5, be placed in closed container in 96 DEG C of stirring gelatinization 60min；
The dextrin cooling that second step obtains step (1), adds isoamylase (Pseudomonas sp, Sigma-
Aldrich), consumption is that every gram of dried starch adds isoamylase 30u, stirring reaction 1 hour at 50 DEG C；
3rd step section pH to 5.5, adds Thermostable α-Amylase (Liquozyme Supra, Novozymes companies),
Consumption is that every gram of dried starch adds Thermostable α-Amylase 10u, and after 1 minute, regulation pH to 2.7 is simultaneously protected stirring reaction at 95 DEG C
Hold 12 minutes enzymes that go out；The IKI mixed solution that addition is configured with water, it is 1mg/ to control concentration of the iodine in mixed solution
ML, it is 10mg/mL to control concentration of the KI in mixed solution；Consumption is that every gram of dried starch adds IKI mixing molten
Liquid 1mL, 70 DEG C of insulated and stirreds 20 minutes, obtains mixture；
Step (3) gained mixture is neutralized and arrives pH6.5 by the 4th step with 0.5mol/L hydrochloric acid, freeze-drying, after crushing
White powder amylose inclusion complexation carrier.After measured (detection method is with embodiment 1), the blue value of products obtained therefrom is
1.06.Stearic acid is embedded using the amylose carrier, the inclusion compound for obtaining shows that (detection method is with implementation after measured
Example 1), stearic content is 9.9% in inclusion compound, and stearic acid embedding rate is 66.3%.
Embodiment 3
First step waxy corn starch disodium hydrogen phosphate-sodium dihydrogen phosphate buffer is made into mass percent concentration and is
8% starch milk, and pH to 5.0 is adjusted, it is placed in closed container in 99 DEG C of stirring gelatinization 40min；
The dextrin cooling that second step obtains step (1), (OPTIMAX L-1000, Jie Neng sections give birth to add Pullulanase
Thing Engineering Co., Ltd), consumption is that every gram of dried starch adds Pullulanase 20u, stirring reaction 2 hours at 65 DEG C；
3rd step section pH to 7.0, adds Thermostable α-Amylase (Liquozyme Supra, Novozymes companies),
Consumption is that every gram of dried starch adds Thermostable α-Amylase 5u, and after 2 minutes, regulation pH to 3.0 is simultaneously protected stirring reaction at 110 DEG C
Hold 15 minutes enzymes that go out；The IKI mixed solution that addition is configured with water, it is 2mg/ to control concentration of the iodine in mixed solution
ML, it is 20mg/mL to control concentration of the KI in mixed solution；Consumption is that every gram of dried starch adds IKI mixing molten
Liquid 0.5mL, 80 DEG C of insulated and stirreds 15 minutes, obtains mixture；
4th step neutralizes to pH6.2 step (3) gained mixture with 0.5mol/L hydrochloric acid, is spray-dried, after crushing
White powder amylose inclusion complexation carrier.After measured (detection method is with embodiment 1), the blue value of products obtained therefrom is
0.96.Stearic acid is embedded using the amylose carrier, the inclusion compound for obtaining shows that (detection method is with implementation after measured
Example 1), stearic content is 9.2% in inclusion compound, and stearic acid embedding rate is 61.6%.
Embodiment 4
It is 10% that first step tapioca disodium hydrogen phosphate-sodium dihydrogen phosphate buffer is made into mass percent concentration
Starch milk, and adjust pH to 6.0, be placed in closed container in 95 DEG C of stirring gelatinization 50min；
The dextrin cooling that second step obtains step (1), adds isoamylase (Pseudomonas sp, Sigma-
Aldrich), consumption is that every gram of dried starch adds isoamylase 10u, stirring reaction 3 hours at 45 DEG C；
3rd step section pH to 6.0, adds mesophilicα-diastase (CLARASE L, Jie Nengke bioengineering Co., Ltd),
Consumption is that every gram of dried starch adds mesophilicα-diastase 2u, and after 5 minutes, regulation pH to 2.7 simultaneously keeps stirring reaction at 60 DEG C
12 minutes enzymes that go out；The IKI mixed solution that addition is configured with water, it is 0.5mg/ to control concentration of the iodine in mixed solution
ML, it is 5mg/mL to control concentration of the KI in mixed solution；Consumption is that every gram of dried starch adds IKI mixed solution
2mL, 60 DEG C of insulated and stirreds 10 minutes, obtains mixture；
Step (3) gained mixture is neutralized and arrives pH6.0 by the 4th step with 0.5mol/L hydrochloric acid, freeze-drying, after crushing
White powder amylose inclusion complexation carrier.After measured (detection method is with embodiment 1), the blue value of products obtained therefrom is
1.12.Stearic acid is embedded using the amylose carrier, the inclusion compound for obtaining shows that (detection method is with implementation after measured
Example 1), stearic content is 10.6% in inclusion compound, and stearic acid embedding rate is 69.1%.
Embodiment 5
First step Waxy wheat starch disodium hydrogen phosphate-sodium dihydrogen phosphate buffer is made into mass percent concentration and is
12% starch milk, and pH to 5.5 is adjusted, it is placed in closed container in 98 DEG C of stirring gelatinization 60min；
The dextrin cooling that second step obtains step (1), (OPTIMAX L-1000, Jie Neng sections give birth to add Pullulanase
Thing Engineering Co., Ltd), consumption is that every gram of dried starch adds Pullulanase 20u, stirring reaction 2 hours at 55 DEG C；
3rd step section pH to 6.5, adds Thermostable α-Amylase (Liquozyme Supra, Novozymes companies),
Consumption is that every gram of dried starch adds high temperature resistant alphalise starch 10u, and after 1 minute, regulation pH to 2.5 is simultaneously protected stirring reaction at 100 DEG C
Hold 10 minutes enzymes that go out；The IKI mixed solution that addition is configured with water, it is 1mg/ to control concentration of the iodine in mixed solution
ML, it is 10mg/mL to control concentration of the KI in mixed solution；Consumption is that every gram of dried starch adds IKI mixing molten
Liquid 1mL, 80 DEG C of insulated and stirreds 20 minutes, obtains mixture；
4th step neutralizes to pH6.5 step (3) gained mixture with 0.5mol/L hydrochloric acid, is spray-dried, after crushing
White powder amylose inclusion complexation carrier.After measured (detection method is with embodiment 1), the blue value of products obtained therefrom is
1.02.Stearic acid is embedded using the amylose carrier, the inclusion compound for obtaining shows that (detection method is with implementation after measured
Example 1), stearic content is 9.7% in inclusion compound, and stearic acid embedding rate is 65%.
Embodiment 6
First step farina disodium hydrogen phosphate-sodium dihydrogen phosphate buffer is made into mass percent concentration and is
10% starch milk, and pH to 4.5 is adjusted, it is placed in closed container in 97 DEG C of stirring gelatinization 50min；
The dextrin cooling that second step obtains step (1), (OPTIMAX L-1000, Jie Neng sections give birth to add Pullulanase
Thing Engineering Co., Ltd), consumption is that every gram of dried starch adds Pullulanase 10u, stirring reaction 3 hours at 60 DEG C；
3rd step section pH to 6.0, adds mesophilicα-diastase (CLARASE L, Jie Nengke bioengineering Co., Ltd),
Consumption is that every gram of dried starch adds mesophilicα-diastase 2u, and after 5 minutes, regulation pH to 2.8 simultaneously keeps stirring reaction at 45 DEG C
15 minutes enzymes that go out；The IKI mixed solution that addition is configured with water, controls concentration of the iodine in mixed solution for 2mg/mL,
It is 20mg/mL to control concentration of the KI in mixed solution；Consumption is that every gram of dried starch adds IKI mixed solution
0.5mL, 70 DEG C of insulated and stirreds 15 minutes, obtains mixture；
Step (3) gained mixture is neutralized and arrives pH6.3 by the 4th step with 0.5mol/L hydrochloric acid, freeze-drying, after crushing
White powder amylose inclusion complexation carrier.After measured (detection method is with embodiment 1), the blue value of products obtained therefrom is
1.18.Stearic acid is embedded using the amylose carrier, the inclusion compound for obtaining shows that (detection method is with implementation after measured
Example 1), stearic content is 11.2% in inclusion compound, and stearic acid embedding rate is 75%.
As described above, you can preferably realize the present invention.
Claims (5)
1. a kind of preparation method of amylose inclusion complexation carrier, it is characterised in that comprise the following steps：
(1) it is 5%~12% that starch material disodium hydrogen phosphate-sodium dihydrogen phosphate buffer is made into mass percent concentration
Starch milk, and pH to 4.5~6.5 is adjusted, it is placed in closed container and 30~60min of gelatinization is stirred at 95~99 DEG C；
(2) the dextrin cooling for obtaining step (1), adds debranching enzyme, consumption be every gram of dried starch add debranching enzyme 10~
30u, stirring reaction 1~3 hour at 45~65 DEG C；
(3) pH to 5.5~7.0 is adjusted, AMS is added, consumption is that every gram of dried starch adds 1~10u of AMS, 45
Stirring reaction 1~10 minute at~110 DEG C；Regulation pH to 2.5~3.0 simultaneously keeps 10~15 minutes enzymes that go out；Addition is configured with water
IKI mixed solution, control concentration of the iodine in mixed solution for 0.5~2mg/mL, control KI is mixing molten
Concentration in liquid is 5~20mg/mL；Consumption is that every gram of dried starch adds IKI mixed solution 0.5~2mL, 60~80 DEG C
Insulated and stirred 10~20 minutes, obtains mixture；
(4) step (3) gained mixture is neutralized to pH 6.0~6.5, amylose inclusion complexation is obtained after dry, pulverize and is carried
Body.
2. the preparation method of amylose inclusion complexation carrier according to claim 1, it is characterised in that：Step (1) institute
It is the one kind in tapioca, cornstarch, farina or wheaten starch to state starch material.
3. the preparation method of amylose inclusion complexation carrier according to claim 1, it is characterised in that：Step (2) is described
Debranching enzyme is Pullulanase or isoamylase.
4. the preparation method of amylose inclusion complexation carrier according to claim 1, it is characterised in that：Institute in step (3)
It is middle temperature or high-temperatureα-amylase to state AMS.
5. the preparation method of amylose inclusion complexation carrier according to claim 1, it is characterised in that：Step (4) is described
Drying be using spray drying or freeze-drying.
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