CN102268417A - Method for extracting malt limit dextrinase - Google Patents
Method for extracting malt limit dextrinase Download PDFInfo
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- CN102268417A CN102268417A CN 201110224011 CN201110224011A CN102268417A CN 102268417 A CN102268417 A CN 102268417A CN 201110224011 CN201110224011 CN 201110224011 CN 201110224011 A CN201110224011 A CN 201110224011A CN 102268417 A CN102268417 A CN 102268417A
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- limit dextrinase
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- malt
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- fructus hordei
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Abstract
The invention discloses a method for extracting malt limit dextrinase, which comprises the following steps: 1) pulverizing dried barley malt with a pulverizer; 2) passing the pulverized malt flour through a 40-80 mesh screen to obtain pretreated malt flour; and 3) adding 0.1 mol/L acetic acid-sodium acetate buffer solution containing 15-25 mmol/L L-cysteine hydrochloride, the pH value of which is 5.0-6.0, regulating the material/liquid ratio to 1:4-1:6 g/mL, putting into a water bath constant-temperature oscillator of which the rotation speed is 80-120 r/min, carrying out separation and extraction at the temperature of 35-45 DEG C for 12-20 hours, and centrifugalizing in a centrifugal machine of which the rotation speed is 3000-5000 r/min, thereby obtaining the malt limit dextrinase. The method can be used for initially separating limit dextrinase, and has the advantages of simple technique, favorable extraction effect and lower cost; and the target enzyme can be maximally dissolved out.
Description
Technical field
The present invention relates to technical field of food biotechnology, be specifically related to a kind of method of extracting the Fructus Hordei Germinatus limit dextrinase.
Background technology
Limit dextrinase (limit dextrinase, EC 3.2.1.4), promptly amylopectin 6-glucose hydrolysis enzyme is a kind of endo-type amylase.α-1,6 glycoside bond in the tapping points such as it can single-minded ground catalysis Propiram, amylopectin, α-limit dextrin, β-limit dextrin.
Because limit dextrinase is to the high degree of specificity of α-1,6 glycoside bond, it has obtained using widely in industry such as food, weaving, washing, particularly in sugaring, starch processing and brewage etc. in the foodstuffs industry and play an important role.In sugar industry, limit dextrinase and the compound use of other amylase can improve conversion coefficient, have important commercial and are worth; In starch industry, the starch after the limit dextrinase effect, good film-forming property has good utility value aspect food product pack; In brewery industry,, can reduce and produce power consumption, production low sugar beer by adding the fermentability that limit dextrinase improves wort.In addition, limit dextrinase also has important value as a kind of lytic enzyme of degraded starch in weaving, medicine, washing industry.At present, Chinese scholars is just in the important application of active development limit dextrinase in related industries.
Limit dextrinase mainly contains two classes: a class derives from microorganism, is called Pullulanase; Another kind ofly derive from plant, be called limit dextrinase or R-enzyme.At present, more external scholars have carried out the research report to the limit dextrinase from plant such as spinach, Chinese sorghum and some bacteriums, extract from the leaf of spinach as people such as Ilka Schindle and to have obtained Pullulanase, but the research of its structure is still waiting further deeply; People such as Cheorl-Ho Kim have isolated the thoroughly I type Pullulanase of hydrolysis Propiram from Thermus caldophilus GK-24; People's separating starch Pullulanases from basophilia bacterium Bacillus sp. KSM-1378 such as Katsutoshi Ara, both the α of hydrolyzable Propiram-1,6 glycoside bond also can act on α in amylopectin, the amylose starch-1,4 glycoside bond.
But food-grade enzyme preparation is strict to originating, and has limited the use of some bacterial origin enzymes.Therefore, the limit dextrinase that extracts from higher plant will have more wide application value.Particularly, limit dextrinase content is abundant relatively in the Fructus Hordei Germinatus, is the good raw material that extracts this enzyme.
Summary of the invention
The object of the present invention is to provide a kind of from Fructus Hordei Germinatus the method for separation extreme dextrinase.This method is extracted environment by regulating, and adds methods such as reductive agent L-halfcystine hydrochloric acid, can extract limit dextrinase to greatest extent, and technology is simple, good separating effect.
Purpose of the present invention mainly realizes by following technological method.
A kind of method of extracting the Fructus Hordei Germinatus limit dextrinase comprises the steps:
(1) dried Fructus Hordei Germinatus is pulverized, crossed 40 ~ 80 purposes sieve, obtain malt meal;
(2) in acetic acid-sodium acetate buffer solution of 0.1mol/L, add reductive agent L-cysteine hydrochloride and obtain extracting buffered soln I;
(3) in malt meal, add extraction buffered soln I in the step (2), place the constant temperature water bath vibrator to carry out lixiviate after, the centrifuging and taking supernatant liquor obtains limit dextrinase in whizzer.
Preferred version is as follows:
The quality of described malt meal is 1:4 ~ 1:6g/mL with the volume ratio of extracting buffered soln I, and the temperature of extraction is 35 ~ 45 ℃, and the time of extraction is 12 ~ 20h.
The pH value of described extraction damping fluid I is 5.0 ~ 6.0.
The concentration of described reductive agent L-cysteine hydrochloride is 15 ~ 25mmol/L.
The rotating speed of described constant temperature water bath vibrator is 80 ~ 120r/min.
The rotating speed of described whizzer is 3000 ~ 5000r/min, and centrifugation time is 10 ~ 15min.
The device of described pulverizing is a pulverizer.
The technology of the present invention analysis of key points:
(1) choosing of raw material: also contain limit dextrinase in the plants such as Chinese sorghum, rice, spinach, wheat, but limit dextrinase content is the abundantest in the Fructus Hordei Germinatus, is best enzyme source.
(2) selection of extraction conditions: very few extracting solution can influence the leaching and the diffusion of enzyme in the Fructus Hordei Germinatus tissue, and too much extracting solution then is unfavorable for concentrating of enzyme liquid, so the malt meal quality is that 1:4 ~ 1:6g/mL is comparatively suitable with extracting buffered soln I volume ratio; The rising of temperature can make the Fructus Hordei Germinatus tissue damaged, is beneficial to the stripping of enzyme, and lower extraction temperature, the stripping enzyme is difficult for losing activity relatively, so it is comparatively reasonable in the time of 35 ~ 45 ℃ to extract temperature; According to the influence to extraction effect of acidity, time and reductive agent consumption, it is preferable to extract pH of buffer value effect in 5.0 ~ 6.0 scopes, and extraction time is better at 12 ~ 20h, and the reductive agent consumption is comparatively suitable at 15 ~ 25 mmol/L.
The present invention has following advantage:
(1) the present invention adopts Fructus Hordei Germinatus as limit dextrin enzyme extraction raw material, and limit dextrinase content is the abundantest in the Fructus Hordei Germinatus, is best enzyme source;
(2) employing the present invention extracts the method for the Fructus Hordei Germinatus limit dextrinase that obtains, the enzyme activity height of gained Fructus Hordei Germinatus limit dextrinase.
Embodiment
In order to understand the present invention better, the present invention is further illustrated below in conjunction with embodiment, but the scope of protection of present invention is not limited to the scope that embodiment represents.
Embodiment 1:
The first step is removed excessive moisture with Fructus Hordei Germinatus dry 24h in 40 ℃ of baking ovens;
Second step adopted the DLFU dry pan to pulverize above-mentioned through the Fructus Hordei Germinatus after the drying treatment, crossed aperture 80 purposes sieve with standby;
The 3rd step was added reductive agent L-cysteine hydrochloride in acetic acid-sodium acetate buffer solution of 0.1mol/L, making its concentration is 20mmol/L, and regulating pH value of buffer solution is 5.0;
The 4th step added the 3rd and goes on foot the extraction damping fluid of being prepared in malt meal, control malt meal concentration is 1:5g/mL, placing rotating speed is the constant temperature water bath vibrator of 100r/min, attemperation is 40 ℃, extract 20h, centrifugal 15min in rotating speed is the whizzer of 3000r/min gets supernatant liquor then, obtain the limit dextrin zyme extract, enzyme activity is 413mU/g.
Embodiment 2:
The first step is removed excessive moisture with Fructus Hordei Germinatus dry 36h in 35 ℃ of baking ovens;
Second step adopted the ZN-08 micromill to pulverize above-mentioned through the Fructus Hordei Germinatus after the drying treatment, crossed aperture 40 purposes sieve with standby;
The 3rd step was added reductive agent L-cysteine hydrochloride in acetic acid-sodium acetate buffer solution of 0.1mol/L, making its solubility is 25mmol/L, and regulating pH value of buffer solution is 5.5;
The 4th step added the 3rd and goes on foot the extraction damping fluid of being prepared in malt meal, control malt meal concentration is 1:6g/mL, placing rotating speed is the constant temperature water bath vibrator of 120r/min, attemperation is 45 ℃, extract 16h, centrifugal 10min in rotating speed is the 5000r/min whizzer gets supernatant liquor then, obtain the limit dextrin zyme extract, enzyme activity is 433mU/g.
Embodiment 3:
The first step is removed excessive moisture with Fructus Hordei Germinatus dry 30h in 38 ℃ of baking ovens;
Second step adopted the grinding cup to pulverize above-mentioned through the Fructus Hordei Germinatus after the drying treatment, crossed aperture 60 purposes sieve with standby;
The 3rd step was added reductive agent L-cysteine hydrochloride in acetic acid-sodium acetate buffer solution of 0.1mol/L, making its solubility is 15mmol/L, and regulating pH value of buffer solution is 6.0;
The 4th step added the 3rd and goes on foot the extraction damping fluid of being prepared in malt meal, control malt meal concentration is 1:4g/mL, placing rotating speed is the constant temperature water bath vibrator of 80r/min, attemperation is 35 ℃, extract 12h, centrifugal 12min in rotating speed is the 4000r/min whizzer gets supernatant liquor then, obtain the limit dextrin zyme extract, enzyme activity is 287mU/g.
Embodiment 4:
The first step is removed excessive moisture with Fructus Hordei Germinatus dry 36h in 35 ℃ of baking ovens;
Second step adopted the DLFU dry pan to pulverize above-mentioned through the Fructus Hordei Germinatus after the drying treatment, crossed aperture 60 purposes sieve with standby;
The 3rd step was added reductive agent L-cysteine hydrochloride in acetic acid-sodium acetate buffer solution of 0.1mol/L, making its solubility is 22mmol/L, and regulating pH value of buffer solution is 5.4;
The 4th step added the 3rd and goes on foot the extraction damping fluid of being prepared in malt meal, control malt meal concentration is 1:5.4 g/mL, placing rotating speed is the constant temperature water bath vibrator of 100 r/min, attemperation is 40 ℃, extract 18h, centrifugal 15min in rotating speed is the whizzer of 3000r/min gets supernatant liquor then, obtain the limit dextrin zyme extract, enzyme activity is 455mU/g.
Claims (6)
1. a method of extracting the Fructus Hordei Germinatus limit dextrinase is characterized in that comprising the steps:
(1) dried Fructus Hordei Germinatus is pulverized, crossed 40 ~ 80 purposes sieve, obtain malt meal;
(2) in acetic acid-sodium acetate buffer solution of 0.1mol/L, add reductive agent L-cysteine hydrochloride and obtain extracting buffered soln I;
(3) in malt meal, add extraction buffered soln I in the step (2), place the constant temperature water bath vibrator to carry out lixiviate after, the centrifuging and taking supernatant liquor obtains limit dextrinase in whizzer.
2. method according to claim 1 is characterized in that the quality of described malt meal and the volume ratio of extracting buffered soln I are 1:4 ~ 1:6g/mL, and the temperature of extraction is 35 ~ 45 ℃, and the time of extraction is 12 ~ 20h.
3. method according to claim 2, the pH value that it is characterized in that described extraction damping fluid I is 5.0 ~ 6.0.
4. method according to claim 1, the concentration that it is characterized in that described reductive agent L-cysteine hydrochloride is 15 ~ 25mmol/L.
5. method according to claim 1, the rotating speed that it is characterized in that described constant temperature water bath vibrator is 80 ~ 120r/min.
6. method according to claim 1, the rotating speed that it is characterized in that described whizzer is 3000 ~ 5000r/min, centrifugation time is 10 ~ 15min.
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| CN 201110224011 CN102268417A (en) | 2011-08-05 | 2011-08-05 | Method for extracting malt limit dextrinase |
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| CN 201110224011 CN102268417A (en) | 2011-08-05 | 2011-08-05 | Method for extracting malt limit dextrinase |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105203367A (en) * | 2015-11-17 | 2015-12-30 | 临沂大学 | Separation method of soluble boron in plants |
| CN113308502A (en) * | 2021-06-10 | 2021-08-27 | 齐鲁工业大学 | Method for preparing corn modified starch by using malt endogenous enzyme |
-
2011
- 2011-08-05 CN CN 201110224011 patent/CN102268417A/en active Pending
Non-Patent Citations (2)
| Title |
|---|
| 《中国海洋大学研究生学位论文》 20091215 李珊 啤酒大麦制麦过程中淀粉酶系及其酶活力的研究 55-57 1-6 , * |
| 《河南农业大学学报》 20070430 艾志录等 小麦芽制备过程中品种间极限糊精酶活力的变化 212-216 1-6 第41卷, 第2期 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105203367A (en) * | 2015-11-17 | 2015-12-30 | 临沂大学 | Separation method of soluble boron in plants |
| CN105203367B (en) * | 2015-11-17 | 2018-02-16 | 临沂大学 | The separation method of soluble boron in a kind of plant |
| CN113308502A (en) * | 2021-06-10 | 2021-08-27 | 齐鲁工业大学 | Method for preparing corn modified starch by using malt endogenous enzyme |
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Application publication date: 20111207 |