CN106699908A - Method for extracting amylose from high amylose corn starch - Google Patents
Method for extracting amylose from high amylose corn starch Download PDFInfo
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- CN106699908A CN106699908A CN201610301739.5A CN201610301739A CN106699908A CN 106699908 A CN106699908 A CN 106699908A CN 201610301739 A CN201610301739 A CN 201610301739A CN 106699908 A CN106699908 A CN 106699908A
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B30/00—Preparation of starch, degraded or non-chemically modified starch, amylose, or amylopectin
- C08B30/20—Amylose or amylopectin
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Abstract
The invention provides a method for extracting amylose from high amylose corn starch. The method comprises the steps that dimethyl sulfoxide (DMSO) is adopted for dispersing amylose and amylopectin into a solution and therefore amylose is extracted; on the basis, the method is improved to adapt to extraction of high amylose corn starch, and finally the high-purity amylose is obtained. By means of the method, a large amount of high-purity amylose can be obtained rapidly from the high amylose corn starch, and the obtained amylose can be adopted as a corn amylose standard product to be applied to corn starch research; compared with the common prior art that commercial potato amylose is adopted as the standard product, the corn starch related research accuracy is greatly improved, and the good application prospect is achieved in corn starch, particularly high amylose corn starch.
Description
Technical field
It is a kind of method extracted from high straight-chain maize and purify amylose the present invention relates to biochemical field.
Background technology
Cornstarch is the important component of corn kernel, accounts for 70% or so of seed dry weight.Ative starch is general by straight chain
Starch, amylopectin and intergrade are grouped into.The starch of separate sources, amylose content is different, in general cereal kind of starch
Amylose content is about 25%；Potato class are about 20%；Beans is about 30-35%.The amylose content of common corn starch exists
27%, and the amylose content of amylomaize is at least more than 50%.Industrially purposes is wider for amylose, can be used to eat
Product process and packaging material manufacture, can be used for water-soluble and biological degradable membrane, it may also be used for medicine and building industry etc. are all
It is multi-field.
But extract that amylose is less efficient, the loss of extraction process starch is serious from common corn starch, and city
Chang Shang maize amyloses supplier is few, and how the extraction amylose of greater efficiency is that limitation corn is straight from cornstarch
One major issue of chain starch application；Often also purity is relatively low for the amylose that laboratory oneself is extracted, and causes to be formed sediment in corn
Potato standard amylose is often needed to use because can not find maize amylose standard items instead, subtracting in powder research field
Few harmful effect to experimental result.
The content of the invention
It is an object of the invention to provide a kind of method that high-purity linear-chain starch is extracted from high straight-chain maize, there is provided one
Plant the method that amylose is separated from corn high amylose starches, obtained on the basis of amylose extracted amount is improve
The method for planting the amylose of higher degree.
Based on above-mentioned purpose, the present invention with reference to a large amount of methods that amylose is extracted from other plant, choose wherein
Part Methods are attempted and improved in high straight-chain maize, and amylose to obtaining carries out purity detecting and recovery rate system
Meter, has obtained a kind of method that high-purity linear-chain starch can be extracted from amylomaize.On this basis, to the party
Method is improved again, and other impurity are removed to greatest extent, improves amylose purity.
Present invention discover that the method that can be used for amylomaize extraction amylose has the retrogradation method after improving（Low turn
Rotating speed fast, high）, the way of distillation and DMSO methods（Dispersion, salting out method）, wherein the amylose that the DMSO dispersion methods after improving are obtained is pure
Spend highest and recovery rate is higher, other 4 methods are substantially not so good as this method.
In order to realize the object of the invention, this utilization DMSO dispersion amyloses and amylopectin from high straight-chain maize
The step of extracting and purify amylose is as follows：
1) extraction of high straight-chain maize seed starch
High straight-chain maize seed 5-15 is taken, after 0.25% sulfurous acid soak 60h, plumule is removed and is planted skin, added water and grind to form
After paste, through 100 μm of nylon net filters to 50 ml centrifuge tubes, 2000 rpm/min are centrifuged 15 min, abandon supernatant, plus 0.5%
NaOH solution 20-40 ml stand 4 h, and 4000 rpm/min are centrifuged 10 min, and NaOH solution repeated washing is colourless to supernatant, sink
Form sediment plus the rpm/min of 15-20 ml acetone 4000 is centrifuged 10 min, it is standby after going to be spontaneously dried in supernatant, fume hood.
2) in amylomaize amylose separation and purifying
Weigh 1-5g high amylose starches and add appropriate DMSO（30-150ml）After be placed in full temperature shaking flask cabinet and slowly stir 24 h,
2000 rpm/min are centrifuged 15 min.Take supernatant and inject 2 times of volume n-butanols, static 24 h, 4000rpm/min centrifugation 10
min.N-butanol washing precipitation 6 times, is transferred in enough water, is heated to being completely dissolved, and appropriate hundred are added when naturally cooling to 60 °C
In phenol（1.6-8g）, room temperature placement 3d, 4000 rpm/min are centrifuged 15 min.Precipitation is transferred in enough water, is heated to completely molten
Solution, adds n-butanol a small amount of after cooling（2-10ml）, indoor standing 24 h, 2000 rpm/min centrifugation 10min, absolute ethyl alcohol washes
Wash 3 times, dry 24 h obtain amylose in being deposited in vacuum freeze drier.
Based on identification amylose purity method it is more, the present invention using the most frequently used blue value method and absorption spectrometry come
Identification amylose purity, using the height of the method statistic recovery rate of amylose quality/ative starch quality.
Blue value is one of important indicator of sign amylose purity, and it embodies the binding ability of starch and iodine.Generally,
Linear polymerization degree is higher, and blue value is higher.Blue value determines the method for using Gilbert, by the acidity for being sufficiently mixed sample
The aqueous solution and Wagner's reagent, after after starch-iodine complex stabilization, using ultraviolet-uisible spectrophotometer to sample 500
Scanned in the range of~700nm, the spectral wavelength at top is designated as maximum absorption wavelength A, determination sample is in maximum absorption wave
The absorbance A of strong point, substitutes into the blue value that formula calculates sample：The solubility of blue value=A*4/ samples（mg/100mL）.Straight chain in theory
Starch blue value is general between 0.8~1.2, and amylose blue value is higher than 0.8, you can think that concentration is higher.This DMSO dispersion sides
The amylose blue value that method is obtained is up to 1.11（Fig. 1）, far above the amylose blue value that other method is obtained, illustrate this method
The amylose of high-purity can be obtained.
The compound formed with iodine using amylose and amylopectin is formed most under ultraviolet specrophotometer scanning
Big absorbing wavelength difference can also carry out amylose purity judgement.Research shows that amylose-Surgidine absworption peak generally exists
Between 600~680nm, and a length of 630nm of the maximum absorption wave of amylose.Fig. 2 illustrates this DMSO methods and extracts the jade for obtaining
The full wavelength scanner spectrogram of rice amylose, maximum absorption wavelength peak value is 628nm, and side demonstrates the straight of this method extraction
Chain purity of starch is high.
It can be seen from the statistics to recovery rate, the recovery rate of the amylose that this method is obtained is compared with other method and fitted
In, this be because good impurity removing effect causes in extraction process, and with using common corn starch extract amylose for,
It is conventional corn with the amylose amount that high amylose starches is obtained as raw material when common starch is identical with high amylose starches usage amount
For 3 times of raw material, (5g conventional corns can extract amylose 0.49g, and 5g high amylose starches can extract 1.36g straight chains shallow lake
Powder), it was demonstrated that with high amylose starches it is the superiority that raw material extracts amylose.
Comprehensive all of analysis of experimental data, the amylose that the present invention is extracted has purity very high, and the present invention is suitable
For extracting amylose in amylomaize, amylose recovery rate is far above common corn starch.Additionally, using horse
Bell potato standard amylose determines cornstarch as standard items may have certain error, therefore, carrying out corn shallow lake
When powder is studied, the amylose standard product of corn source should be used.In sum, the method for being provided using the present invention, can make
The amylose of standby high-purity, can as maize amylose standard items, for the correlative study of corn kernel starch, while
The speed that amylose puts goods on the market can be accelerated.
Brief description of the drawings
Fig. 1 is the inventive method DMSO dispersion methods and the blue value analysis of other extraction amylose methods.
Fig. 2 is the absorption spectrum curve of the inventive method.
Claims (6)
1. a kind of amylomaize extracts the purification process of amylose, it is characterised in that comprise the following steps:
1) extraction of high straight-chain maize seed starch
By high straight-chain maize seed（5-15）60h is soaked with the sulfurous acid solution of volume fraction 0.25%, is then cut with a knife and is removed
Plumule and kind of a skin is peelled off, plus after a small amount of water grinds to form paste, through 100 μm of nylon net filters to 50 mL centrifuge tubes, 2000 rpm/
Min is centrifuged 15 min, abandons supernatant, after the mL of NaOH solution 40 for plus 0.2% is well mixed, stands 4 h, then in 4000 rpm/min
10 min are centrifuged, 0.2% NaOH solution repeated washing is colourless to supernatant, precipitation plus the rpm/min of 15 mL acetone 4000 are centrifuged
10 min, it is standby after going to be spontaneously dried in supernatant, fume hood
2) in amylomaize amylose separation and purifying
The high amylose starches for weighing 1-5 g extractions adds the beaker of 30-150mLDMSO for configure, placement after stirring
After slowly stirring 24 h with the speed of 120r/min in full temperature shaking flask cabinet, mixed liquor is individually placed in 50ml centrifuge tubes with 2000
Rpm/min is centrifuged 15-20min, takes the supernatant after centrifugation and injects 2 times of volume n-butanols, static 24 h, then in 4000rpm/
Min is centrifuged 10 min, after n-butanol washing precipitates 6 times, is transferred to enough water（30-40ml）In, it is heated to being completely dissolved, it is naturally cold
Add 1.6-8 g thymols when but to 60 °C, room temperature places 3 d, and 4000 rpm/min are centrifuged 15 min, and precipitation is transferred to enough water
In (20-30mL), it is heated to being completely dissolved, n-butanol 10mL, indoor standing 24 h, 2000 rpm/min centrifugation is added after cooling
10min, 3 removing n-butanols of absolute ethanol washing, dry 24 h obtain amylose in being deposited in vacuum freeze drier.
2. the maize amylose that according to claim 1 prepared by method is high-purity linear-chain starch, is that one kind prepares corn
The method of amylose standard product.
3. it is according to claim 1 from amylomaize extract amylose method, it is characterised in that：It is described
Method from amylomaize suitable for extracting amylose.
4. method according to claim 1, it is characterised in that：Methods described needs 5g starch in the DMSO solution of q.s
In（30-150ml）Stir, and DMSO/ starch mixed liquor needs that 15-20min is centrifuged with 2000rpm/min.
5. method according to claim 1, it is characterised in that：, it is necessary to use during with n-butanol separation straight chain/amylopectin
2 times of n-butanols of amount supernatant, and 10min is centrifuged under conditions of 4000rpm/min, and precipitate 6 times.
6. method according to claim 1, it is characterised in that：The amylose that methods described is obtained need to be dry in vacuum refrigeration
Could be dried under the conditions of dry.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610301739.5A CN106699908A (en) | 2016-05-09 | 2016-05-09 | Method for extracting amylose from high amylose corn starch |
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| CN201610301739.5A CN106699908A (en) | 2016-05-09 | 2016-05-09 | Method for extracting amylose from high amylose corn starch |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111004332A (en) * | 2019-12-13 | 2020-04-14 | 山东省农作物种质资源中心 | Method for separating and purifying amylose and amylopectin in wheat |
| CN112345323A (en) * | 2020-10-31 | 2021-02-09 | 中国水稻研究所 | Method for developing standard substance of amylose content of rice |
| CN113234177A (en) * | 2021-06-18 | 2021-08-10 | 山东福洋生物科技股份有限公司 | Production method of high-amylose corn starch |
| CN114989316A (en) * | 2022-06-01 | 2022-09-02 | 山东省农业科学院 | Method for obtaining high amylose starch from cashew kernels |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1363610A (en) * | 2001-11-16 | 2002-08-14 | 吉林省轻工业设计研究院 | Improved immersion process in preparing straight-chain corn starch |
| CN104926949A (en) * | 2015-07-10 | 2015-09-23 | 华东理工大学 | Preparation method for high purity amylose |
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2016
- 2016-05-09 CN CN201610301739.5A patent/CN106699908A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1363610A (en) * | 2001-11-16 | 2002-08-14 | 吉林省轻工业设计研究院 | Improved immersion process in preparing straight-chain corn starch |
| CN104926949A (en) * | 2015-07-10 | 2015-09-23 | 华东理工大学 | Preparation method for high purity amylose |
Non-Patent Citations (2)
| Title |
|---|
| 李彬等: ""玉米直链淀粉、支链淀粉的分离、表征及浮选应用"", 《矿产保护与利用》 * |
| 李志伟等: ""高直链玉米淀粉的理化特性研究"", 《西北农林科技大学学报（自然科学版）》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111004332A (en) * | 2019-12-13 | 2020-04-14 | 山东省农作物种质资源中心 | Method for separating and purifying amylose and amylopectin in wheat |
| CN112345323A (en) * | 2020-10-31 | 2021-02-09 | 中国水稻研究所 | Method for developing standard substance of amylose content of rice |
| CN113234177A (en) * | 2021-06-18 | 2021-08-10 | 山东福洋生物科技股份有限公司 | Production method of high-amylose corn starch |
| CN114989316A (en) * | 2022-06-01 | 2022-09-02 | 山东省农业科学院 | Method for obtaining high amylose starch from cashew kernels |
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Application publication date: 20170524 |