CN104263781A - Ultrasonic-assisted double-enzyme hydrolysis technology of starch - Google Patents
Ultrasonic-assisted double-enzyme hydrolysis technology of starch Download PDFInfo
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- CN104263781A CN104263781A CN201410511137.3A CN201410511137A CN104263781A CN 104263781 A CN104263781 A CN 104263781A CN 201410511137 A CN201410511137 A CN 201410511137A CN 104263781 A CN104263781 A CN 104263781A
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Abstract
The invention discloses an ultrasonic-assisted double-enzyme hydrolysis technology of starch and belongs to the technical field of enzymolysis of starch. The ultrasonic-assisted double-enzyme hydrolysis of starch is especially applied in the invention. According to the method, ultrasonic wave is added in starch liquefaction and glucoamylase hydrolysis processes, under the conditions that the ultrasonic power is 100W, the ultrasonic time is 10 minutes and the concentration of starch milk is 20%, the liquification value of starch milk and DE (Dextrose Equivalent) value are respectively improved from 19.89mg/mL and 82.06%, which are obtained from starch milk untreated by ultrasonic wave, to 30.67mg/mL and 94.30%. Since starch particles are subjected to ultrasonic treatment, dents and cracks on the surface of the starch particles are significantly increased, the crystalline structure of starch is destroyed, the infrared crystallinity index is decreased, the starch branched structure is destroyed, the content of amylase is increased and thus the solubility is improved by 246.8%. Furthermore, due to the activation effect of ultrasonic treatment on the enzyme, the activity of alpha-amylase is improved by 15.29%.
Description
Technical field
The present invention relates to a kind of amylorrhexis technical field, particularly relate to a kind of ultrasound-assisted enzymolysis technique.
Background technology
Starch (starch) is a kind of superpolymer of natural hypocrystalline grain pattern, there is stuctures and properties defect, crystallizing field molecular arrangement is tight, water, enzyme and most chemical reagent is caused not easily to touch molecule in crystallizing field, thus show be insoluble to cold water, the easy aging dehydration of starch paste, lack emulsifying power, and starch paste is unstable under acid, heat and shear action, the phenomenons such as chemical reaction efficiency is lower.
Along with the development of starch industry, the status of starch saccharification processes in the industries such as sugaring, alcohol, organic acid fermentation becomes more and more important.
The starch utilization ratio of current existence is low, and is adopt the problems such as single enzyme hydrolysis mostly.These problems have become the bottleneck of starch industry development, find one of effective way focus becoming research improving raw starch transformation efficiency.
Ultrasonic wave is the sound wave of frequency higher than 20 KHz, is a kind of elastic mechanical ripple, is widely used in recent years in the fields such as physics, biology, chemistry.Ultrasonication can make the high molecular morphological structure of biomass and ultrastructure generation considerable change, is conducive to the accessibility and the chemical reactivity that improve enzyme.Yet there are no and ultrasonic assistant α-amylase and saccharifying enzyme double-enzyme method hydrolyzed starch are reported with the research improving its transformation efficiency.
Summary of the invention
The invention provides a kind of enzyme solution of starch, namely by while adding α-amylase (Thermostable α-Amylase 10 U/g raw material) and saccharifying enzyme (saccharifying enzyme 300 U/g raw material) in addition ultrasonic wave starch is hydrolyzed.Overcome the problem that starch utilization ratio is low.
For achieving the above object, this experiment have employed following technical scheme:
(1) starch milk pre-treatment: the starch and the distilled water that weigh certain mass, sizes mixing into the starch milk of mass concentration 20%.Get 200 mL starch milks in 300 mL beakers, 20 DEG C of water bath with thermostatic control agitators put into by beaker, ultrasonic variable amplitude bar inserted below starch milk liquid level 2 cm, magnetic agitation rotating speed 50%, ultrasonic (working hour/intermittent time=2 s/4 s) process 20 min.
(2) liquefaction process supersound process: the starch and the distilled water that weigh certain mass, sizes mixing into the starch milk of mass concentration 20%.Get 200 mL starch milks in 300 mL beakers, pH is adjusted to be 2.2,95 DEG C of water bath with thermostatic control agitators put into by beaker, ultrasonic variable amplitude bar is inserted below starch milk liquid level 2 cm, magnetic agitation rotating speed 50%, ultrasonic (working hour/intermittent time=2 s/4 s), ultrasonic time 10 min, ultrasonic power 100 W, total liquefying time 50 min.After having liquefied, cooling, go out enzyme 30 min.
(3) saccharifying supersound process: the starch and the distilled water that weigh certain mass, sizes mixing into the starch milk of mass concentration 20%.Get 200 mL starch milks in 300 mL beakers, after having liquefied, ultrasonic variable amplitude bar is inserted below starch milk liquid level 2 cm, magnetic agitation rotating speed 50%, ultrasonic (working hour/intermittent time=2 s/4 s), ultrasonic power 100 W, auxiliary saccharification 18 min.
Preparation method of the present invention has following features:
(1) starch milk of ultrasonication is compared with the starch milk not adding ultrasonication, and its liquefaction value is raised to 38.86 mg/mL from 22.67 mg/mL, and DE value is raised to 94.30% from 82.70%.Its reason may be the effect of ultrasonic wave to starch milk
(2) ul-trasonic irradiation destroys the water bundle layer of starch particle surface, makes moisture infiltrate starch granules, is conducive to the hydrolysis of amylase to starch
(3) shearing force that ultrasonic wave " hole effect " produces cuts off the long-chain of starch, is conducive to the enzymolysis of starch
(4) free radical that ultrasonic wave produces can attack starch molecule, and cause the fracture of Isosorbide-5-Nitrae-glycosidic link, starch long-chain exposes a large amount of non reducing end, for enzymolysis provides more substrate
(5) hyperacoustic mechanical agitation make whole reaction system evenly, the contact frequency of substrate and zymophore increases
(6) between the water and air that causes of ultrasonic air cavitation, interfacial area increases, and has upset enzyme surrounding molecules environment, as hydrogen bond and hydrophobic interaction, has caused the change of enzyme texture image
(7) ultrasonic wave generation shearing force and shockwave cause the change of enzyme molecular structure, show the change that enzyme is lived.
(8) after starch milk ultrasonication, enzymolysis has more efficient advantage than simple enzyme solution.
(9) ultrasonic assistant liquefaction, is conducive to the degraded of starch polymer, can improve, for saccharifying provides sufficient substrate the ratio of enzymatic starch.
(10) along with the increase of ultrasonic power, the starch granules that surface weathers increases, and cavity becomes large, and deepen, crackle increases, and the granule interior appearance that weathers is caved in even defect.The outer crystallizing field of starch granules is destroyed, chemical reagent can fast with the effect of starch granules internal structure, be favourable to promotion starch chemical reactivity.
(11) starch crystals district is damaged in ultrasonication process, the high pressure that cavitation bubble produces and local torrent have enough shearing forces to break polymeric chain, cause the fracture of the even whole starch granules in starch crystals district, this is also the major cause that amylase activity increases.
(12) the outer field firm crystallizing field of starch receives destruction under ultrasound environments, and water molecules and internal starch molecule contacts are more prone to, and associating between starch and water molecules increases, and starch dissolution degree increases.
Embodiment 1
(1) weigh starch and the distilled water of certain mass, size mixing into the starch milk of mass concentration 20%.Get 200 mL starch milks in 300 mL beakers, adjust pH to be 5.6 ± 0.2,95 DEG C of water bath with thermostatic control agitators put into by beaker.
(2) ultrasonic variable amplitude bar is inserted below starch milk liquid level 2 cm, magnetic agitation rotating speed 50%, ultrasonic (working hour/intermittent time=2 s/4 s) auxiliary liquefaction 10min, ultrasonic power 100 W, total liquefying time 50 min.After having liquefied, cooling, go out enzyme 30min.
(3) mensuration of liquefaction value adopts DNS method to carry out.And calculate liquefaction value according to formula (1)
（1）
In formula
a-light absorption value,
n-extension rate.
Embodiment 2
(1) weigh starch and the distilled water of certain mass, size mixing into the starch milk of quality dense 20%.Get 200 mL starch milks in 300 mL beakers.
(2) then adjust pH4.2 ~ 4.4 with 1 mol/L NaOH after having liquefied, add saccharifying enzyme (saccharifying enzyme 300 U/g raw material), put into 60 DEG C of water bath with thermostatic control agitators and carry out saccharification react.
(3) ultrasonic variable amplitude bar is inserted below starch milk liquid level 2 cm, magnetic agitation rotating speed 50%, ultrasonic (working hour/intermittent time=2 s/4 s) ultrasonic power 100 W, auxiliary saccharification 18min, after saccharification completes, cooling, surveying its DE value is 87.31%, describes the saccharification that degraded that ultrasonic wave is conducive to starch polymer can improve starch milk to a certain extent.
(4) DE pH-value determination pH is with reference to GB/T 22428.1-2008 starch hydrolysis production reducing power and glucose equivalent measuring method.
Claims (6)
1. a ultrasonic assistant starch double-enzyme hydrolysis technology, is characterized in that equal supersound process in starch milk pre-treatment, liquefaction process and saccharifying.
2. according to claim 1, starch milk pre-treatment is characterized in that the starch and the distilled water that weigh certain mass, sizes mixing into the starch milk of mass concentration 10% ~ 40%;
Get 200 mL starch milks in 300 mL beakers, 20 DEG C of water bath with thermostatic control agitators put into by beaker, ultrasonic variable amplitude bar inserted below starch milk liquid level 2 cm, magnetic agitation rotating speed 50%, ultrasonic (working hour/intermittent time=2 s/4 s) process certain hour.
3. according to claim 1, liquefaction process supersound process is characterized in that the starch and the distilled water that weigh certain mass, sizes mixing into the starch milk of certain mass concentration;
Get 200 mL starch milks in 300 mL beakers, pH is adjusted to be 2.2,95 DEG C of water bath with thermostatic control agitators put into by beaker, ultrasonic variable amplitude bar is inserted below starch milk liquid level 2 cm, magnetic agitation rotating speed 50%, ultrasonic (working hour/intermittent time=2 s/4 s) auxiliary liquefaction certain hour, after having liquefied, cooling, go out enzyme 30 min.
4. according to claim 3, wherein, liquefaction condition is ultrasonic time 10 min, ultrasonic power 100 W, starch concentration 20%, total liquefying time 50 min.
5. according to claim 1, saccharifying supersound process is characterized in that the starch and the distilled water that weigh certain mass, sizes mixing into the starch milk of certain mass concentration;
Get 200 mL starch milks in 300 mL beakers, after having liquefied, ultrasonic variable amplitude bar is inserted below starch milk liquid level 2 cm, magnetic agitation rotating speed 50%, ultrasonic (working hour/intermittent time=2 s/4 s) auxiliary saccharification certain hour.
6. according to claim 5, wherein, saccharification condition is ultrasonic time 10 min, ultrasonic power 100 W, starch concentration 20%.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104862375A (en) * | 2015-06-12 | 2015-08-26 | 江南大学 | Application of dextrin molecular weight characteristics in detecting liquefication effect of starch |
| CN106635580A (en) * | 2016-11-03 | 2017-05-10 | 张静 | Production method of syrup special for sorghum-brewed beer |
| CN107337740A (en) * | 2017-07-24 | 2017-11-10 | 江苏大学 | A kind of method of arrowhead starch double-frequency ultrasound wavefront modifier |
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| US20050136520A1 (en) * | 2003-10-03 | 2005-06-23 | Kinley Michael T. | Biomass conversion to alcohol using ultrasonic energy |
| CN102030630A (en) * | 2010-11-16 | 2011-04-27 | 湖北丹江口丹澳医药化工有限公司 | Method for producing industrial grade sodium gluconate or calcium gluconate from yellow ginger starch |
| CN103981240A (en) * | 2014-05-13 | 2014-08-13 | 江南大学 | Method of increasing the saccharification reaction efficiency of high-concentration starch |
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2014
- 2014-09-29 CN CN201410511137.3A patent/CN104263781A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| US20050136520A1 (en) * | 2003-10-03 | 2005-06-23 | Kinley Michael T. | Biomass conversion to alcohol using ultrasonic energy |
| CN102030630A (en) * | 2010-11-16 | 2011-04-27 | 湖北丹江口丹澳医药化工有限公司 | Method for producing industrial grade sodium gluconate or calcium gluconate from yellow ginger starch |
| CN103981240A (en) * | 2014-05-13 | 2014-08-13 | 江南大学 | Method of increasing the saccharification reaction efficiency of high-concentration starch |
Non-Patent Citations (3)
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| STEPHEN BARTON ET AL.: "The effects of ultrasound on the activities of some glycosidase enzymes of industrial importance", 《ENZYME AND MICROBIAL TECHNOLOGY》 * |
| 孙俊良等: "超声波辅助酶解淀粉制备糊精的工艺研究", 《食品科学》 * |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104862375A (en) * | 2015-06-12 | 2015-08-26 | 江南大学 | Application of dextrin molecular weight characteristics in detecting liquefication effect of starch |
| CN104862375B (en) * | 2015-06-12 | 2018-06-19 | 江南大学 | Application of the dextrin molecular weight characteristic in detection starch liquefacation effect |
| CN106635580A (en) * | 2016-11-03 | 2017-05-10 | 张静 | Production method of syrup special for sorghum-brewed beer |
| CN107337740A (en) * | 2017-07-24 | 2017-11-10 | 江苏大学 | A kind of method of arrowhead starch double-frequency ultrasound wavefront modifier |
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