CN101892220B - Method for improving activity of malt amylase - Google Patents
Method for improving activity of malt amylase Download PDFInfo
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- CN101892220B CN101892220B CN2010102299861A CN201010229986A CN101892220B CN 101892220 B CN101892220 B CN 101892220B CN 2010102299861 A CN2010102299861 A CN 2010102299861A CN 201010229986 A CN201010229986 A CN 201010229986A CN 101892220 B CN101892220 B CN 101892220B
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- amylase
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- malt amylase
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- GXCLVBGFBYZDAG-UHFFFAOYSA-N N-[2-(1H-indol-3-yl)ethyl]-N-methylprop-2-en-1-amine Chemical compound CN(CCC1=CNC2=C1C=CC=C2)CC=C GXCLVBGFBYZDAG-UHFFFAOYSA-N 0.000 title claims abstract description 37
- 230000000694 effects Effects 0.000 title claims abstract description 33
- 239000004382 Amylase Substances 0.000 title claims abstract description 31
- 102000013142 Amylases Human genes 0.000 title claims abstract description 31
- 108010065511 Amylases Proteins 0.000 title claims abstract description 31
- 235000019418 amylase Nutrition 0.000 title claims abstract description 31
- 238000000034 method Methods 0.000 title claims abstract description 28
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims abstract description 16
- 239000000287 crude extract Substances 0.000 claims abstract description 13
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims abstract description 8
- 235000012054 meals Nutrition 0.000 claims description 11
- 230000005855 radiation Effects 0.000 claims description 8
- 239000000872 buffer Substances 0.000 claims description 6
- 239000006228 supernatant Substances 0.000 claims description 6
- 238000006243 chemical reaction Methods 0.000 claims description 3
- 239000008055 phosphate buffer solution Substances 0.000 claims description 3
- 238000010298 pulverizing process Methods 0.000 claims description 2
- 229920002472 Starch Polymers 0.000 abstract description 10
- 235000019698 starch Nutrition 0.000 abstract description 10
- 239000008107 starch Substances 0.000 abstract description 10
- 235000013405 beer Nutrition 0.000 abstract description 8
- 235000007340 Hordeum vulgare Nutrition 0.000 abstract description 3
- 230000007062 hydrolysis Effects 0.000 abstract description 3
- 238000006460 hydrolysis reaction Methods 0.000 abstract description 3
- 238000004519 manufacturing process Methods 0.000 abstract description 3
- 239000000243 solution Substances 0.000 abstract description 3
- 230000008901 benefit Effects 0.000 abstract description 2
- 241000209219 Hordeum Species 0.000 abstract 2
- 239000007853 buffer solution Substances 0.000 abstract 1
- 230000001678 irradiating effect Effects 0.000 abstract 1
- 239000000843 powder Substances 0.000 abstract 1
- 102000004190 Enzymes Human genes 0.000 description 18
- 108090000790 Enzymes Proteins 0.000 description 18
- 230000008569 process Effects 0.000 description 7
- 238000002604 ultrasonography Methods 0.000 description 6
- 239000002253 acid Substances 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 238000001035 drying Methods 0.000 description 4
- 238000012216 screening Methods 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 238000003805 vibration mixing Methods 0.000 description 4
- 239000003814 drug Substances 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 238000000855 fermentation Methods 0.000 description 3
- 230000004151 fermentation Effects 0.000 description 3
- 241000209140 Triticum Species 0.000 description 2
- 235000021307 Triticum Nutrition 0.000 description 2
- 230000004071 biological effect Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000002478 diastatic effect Effects 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000005360 mashing Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 238000003672 processing method Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 1
- 229920002527 Glycogen Polymers 0.000 description 1
- 240000005979 Hordeum vulgare Species 0.000 description 1
- 206010020843 Hyperthermia Diseases 0.000 description 1
- 244000017020 Ipomoea batatas Species 0.000 description 1
- 235000002678 Ipomoea batatas Nutrition 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 244000061456 Solanum tuberosum Species 0.000 description 1
- 235000002595 Solanum tuberosum Nutrition 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000005844 autocatalytic reaction Methods 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 230000002925 chemical effect Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000006854 communication Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 230000005669 field effect Effects 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 229940096919 glycogen Drugs 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 230000036031 hyperthermia Effects 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 230000000644 propagated effect Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000010008 shearing Methods 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000009923 sugaring Methods 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 239000004753 textile Substances 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000009941 weaving Methods 0.000 description 1
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Enzymes And Modification Thereof (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The invention discloses a method for improving the activity of malt amylase. The method comprises the following steps: crushing the dried barley malt by using a crusher, adding a phosphoric acid buffer solution with the pH value of 5.8 into the barley malt powder, standing at room temperature for 40-60 min, and centrifuging to obtain a crude extract of malt amylase; irradiating the coarse malt amylase extracting solution by using ultrasonic waves under the ultrasonic conditions: the fixed ultrasonic frequency is 20kHz, the ultrasonic power is 120-160W, the total ultrasonic irradiation time is 10-20 min, the ultrasonic irradiation time is 3s, the time is 1 s-9 s, the ultrasonic irradiation time is 10-20 min, and the temperature of an ultrasonic system is less than 4 ℃. The method can obviously improve the activity of the malt amylase by 20-30 percent, thereby improving the hydrolysis rate of the starch, effectively reducing the production cost of beer brewing, saving energy and resources and having remarkable social and economic benefits.
Description
Technical field
The present invention relates to technical field of food biotechnology, be specifically related to a kind of ultrasonic processing method that improves activity of malt amylase.
Background technology
Glycase is the enzyme general name of hydrolyzed starch and glycogen, and they extensively are present in plant (like barley, potato, sweet potato etc.), animal and some mikrobes, its brewage, industry such as grain processing, textiles and medicine has important effect.Particularly the application in beer prodn is the focus of studying both at home and abroad to glycase always as saccharifying agent.
Fructus Hordei Germinatus is the main raw material of brewing industry, is the guarantee of brewing industry development.The main reserve substance of Fructus Hordei Germinatus is a starch, and it generates the small molecules fermentable sugar through a series of diastatic actions can be by the yeast utilization.Therefore, in the beer production, the height of activity of malt amylase directly determines the content of fermentable sugar in the wheat juice, influences the beer fermentation degree, thereby influences the yield and quality of beer.At present, improving the beer fermentation degree is the technological difficulties of brewing industry.Both at home and abroad some beer enterprises mainly improve brewing quality through plus enzyme, and this method instant effect and effect are remarkable, but the cost issues limit its further application.In addition, the content through fermentable sugar in adjustment Mashing process or the outer sugaring method raising wheat juice also is limited.Therefore, seeking a kind of cheapness, new and high technology efficient, environmental protection, fundamentally to improve amylase activity be present problem demanding prompt solution.
UW is a kind of elastic mechanical ripple in the material medium, and its range of frequency is 1.6 * 10
4Hz～10
9Hz.When UW is propagated, produce a series of special effects in liquid, like cavatition, calorifics effect, chemical effect and biological effect etc.Simultaneously, UW has efficiently, characteristics such as cheap, pollution-free, and these characteristics make UW have a wide range of applications in food-processing, medicine, chemical industry and biotechnology field.
For biomacromolecule protein; When putting on solution, UW produces cavitation effect (growth of bubble and collapse suddenly); Temperature-rise effect (absorption ultrasonic energy), radical effect, the powerful stirring and shearing; Miniflow effects etc. can change proteinic secondary, three grades and high-grade conformation more effectively.The speed of response of enzyme depends primarily on two factors: the conformation of mass-transfer efficiency and enzyme molecule.UW acts on enzyme and divides the period of the day from 11 p.m. to 1 a.m, and the energy of release can cause the conformation of enzyme molecule and structure to change, thereby has influence on the variation of catalytic activity.Therefore, the application of ultrasonic technique on enzyme engineering will bring vast potential for future development for foodstuffs industry.
Summary of the invention
The object of the present invention is to provide a kind of method that improves activity of malt amylase.This method can obviously improve activity of malt amylase, improves the hydrolysis rate of starch, reduces enterprise cost, and save energy and resource are had significance.
The object of the invention is mainly realized through ultrasonic processing method.Main technique methods is:
A kind of method that improves the Fructus Hordei Germinatus amylase activity comprises the steps:
(1) dried Fructus Hordei Germinatus is pulverized, crossed 40～80 purposes sieve, obtain malt meal;
(2) in malt meal, add phosphate buffer solution, centrifugal behind placement 40～60min, get supernatant, obtain the malt amylase crude extract;
(3) the malt amylase crude extract is carried out the intermittent type ultrasonic irradiation, ultrasonic irradiation 3s stops 1s～9s, UW mutual radiation 10～20min, and said ultrasonic frequency is 20kHz, ultrasonic power 120～160W.
Said centrifugal rotation speed is 1500～3000r/min, and the time is 10～15min.
The slow molten mass volume ratio towards liquid of said malt meal and phosphoric acid is 1: 15～1: 10g/ml.
The pH=5.8 of said phosphoric acid buffer.
The device of said pulverizing is a kibbler.
Temperature of reaction system is lower than 4 ℃ when described ultrasonic power 120～160W, said ultrasound wave irradiation, and to guarantee the righttest performance of effect of ultrasound to diastatic action, that avoids material simultaneously crosses heat-flash effect and localized hyperthermia.
Principle of the present invention: UW is a kind of physical wave, and in its communication process, the energy that UW has discharges gradually and acts on the physics particle in its propagation medium, thereby makes these particles that corresponding physical change and chemical transformation take place.Enzyme is a biomacromolecule, has comparatively stable conformation, and the resonable degree of enzyme texture image is depended in the performance of enzyme molecular biological activity fully.Divide the period of the day from 11 p.m. to 1 a.m when ultrasonic field acts on enzyme, the energy that UW discharges can cause the variation of enzyme texture image.Reasonably conformational change can make the ultrastructure of enzyme molecule have more flexibility, makes that its space structure is preferably to be combined with substrate, thereby brings into play higher catalysis.The sound field effect enzyme that intensity is lower divides the period of the day from 11 p.m. to 1 a.m, and the energy that is discharged can make enzyme texture image be tending towards rationally, thereby it is active to improve autocatalysis.But stronger supersound process can cause enzyme texture image to destroy, and influences the active site of enzyme, and then enzyme activity is descended even inactivation.
The present invention compared with prior art has following advantage:
(1) the present invention improves activity of malt amylase with UW as main means, has solved the major technique defective that activity of malt amylase is not enough, the starch hydrolysis efficiency is not high, the beer fermentation degree is low;
(2) simple, cheap, the environmental protection of the present invention's technology can reduce enterprise's raw materials cost, shorten Mashing process, and save energy and resource have remarkable economic efficiency;
(3) the present invention also can be used for other industries such as starch syrup production, weaving and medicine, is with a wide range of applications.
Embodiment
In order to understand the present invention better, below in conjunction with embodiment the present invention is further described, but the scope that the present invention requires to protect is not limited to the scope that embodiment representes.
Embodiment 1:
The first step is Fructus Hordei Germinatus dry 24h in 40 ℃ of baking ovens;
Second step adopted DL F USB flash disk formula kibbler to pulverize above-mentioned through the Fructus Hordei Germinatus after the drying treatment, crossed aperture 80 purposes sieve with subsequent use;
Interpolation pH is 5.8 phosphoric acid buffer in the malt meal of the 3rd step after the screening of second step is handled, and malt meal concentration is controlled to be 1: 13g/ml, vibration mixing; Room temperature held 40min; Centrifugal 10min under 3000r/min gets supernatant then, obtains the malt amylase crude extract;
The 4th step utilized the JY92-2D ultrasonic cell disruptor that the malt amylase crude extract is carried out supersound process, and ultrasound condition is: ultrasonic frequency 20kHz, ultrasonic power 120W; 1: 1 (ultrasonic time 3s of pulse mode; Stop 3s), UW radiation total time 15min, 3 ℃ of ultrasonic system temperatures.Reach 1238.932U/g through DNS (3, the 5-dinitrosalicylic acid) its amylase activity of method test analysis, improved 27.17% than Fructus Hordei Germinatus ative starch enzymic activity.
Embodiment 2:
The first step is removed excessive moisture with Fructus Hordei Germinatus dry 36h in 35 ℃ of baking ovens;
Second step adopted ZN-08 laboratory micromill to pulverize above-mentioned through the Fructus Hordei Germinatus after the drying treatment, crossed aperture 40 purposes sieve with subsequent use;
Interpolation pH is 5.8 phosphoric acid buffer in the malt meal of the 3rd step after the screening of second step is handled, and concentration is controlled to be 1: 10g/ml, and the vibration mixing, room temperature held 60min, the centrifugal 15min of 1500r/min gets supernatant then, obtains the malt amylase crude extract;
The 4th step utilized the JY92-2D ultrasonic cell disruptor that the malt amylase crude extract is carried out supersound process, and ultrasound condition is: ultrasonic frequency 20kHz, ultrasonic power 160W; 3: 1 (UW radiated time 3s of pulse mode; Stop 1s), UW radiation total time 10min, 2 ℃ of ultrasonic system temperatures.Reach 1202.596U/g through DNS (3, the 5-dinitrosalicylic acid) its amylase activity of method test analysis, improved 23.44% than Fructus Hordei Germinatus ative starch enzymic activity.
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 subsequent use;
Interpolation pH is 5.8 phosphoric acid buffer in the malt meal of the 3rd step after the screening of second step is handled, and concentration is controlled to be 1: 15g/ml, and the vibration mixing, room temperature held 50min, the centrifugal 15min of 1500r/min gets supernatant then, obtains the malt amylase crude extract;
The 4th step utilized the BILON-1200Y ultrasonic cell disruptor that the malt amylase crude extract is carried out supersound process; Ultrasound condition is: fixedly ultrasonic frequency is 20kHz; Ultrasonic power 120W, UW radiation total time 20min, 1: 3 (ultrasonic radiation time 3s of pulse mode; Stop 9s), 3 ℃ of ultrasonic system temperatures.Reach 1209.982U/g through DNS (3, the 5-dinitrosalicylic acid) its amylase activity of method test analysis, improved 24.20% than Fructus Hordei Germinatus ative starch enzymic activity.
Embodiment 4:
The first step is removed excessive moisture with Fructus Hordei Germinatus dry 36h in 35 ℃ of baking ovens.
Second step adopted ZN-08 laboratory micromill to pulverize above-mentioned through the Fructus Hordei Germinatus after the drying treatment, crossed aperture 80 purposes sieve with subsequent use;
Interpolation pH is 5.8 phosphoric acid buffer in the malt meal of the 3rd step after the screening of second step is handled, and concentration is controlled to be 1: 12g/ml, and the vibration mixing, room temperature held 60min, the centrifugal 10min of 3000r/min gets supernatant then, obtains the malt amylase crude extract;
The 3rd step utilized the BILON-1200Y ultrasonic cell disruptor that the malt amylase crude extract is carried out supersound process; Ultrasound condition is: fixedly ultrasonic frequency is 20kHz; Ultrasonic power 140W, UW radiation total time 20min, 3: 2 (ultrasonic radiation time 3s of pulse mode; Stop 2s), 2 ℃ of ultrasonic system temperatures.Reach 1218.389U/g through DNS (3, the 5-dinitrosalicylic acid) its amylase activity of method test analysis, improved 25.07% than Fructus Hordei Germinatus ative starch enzymic activity.
Claims (3)
1. a method that improves the Fructus Hordei Germinatus amylase activity is characterized in that, comprises the steps:
(1) dried Fructus Hordei Germinatus is pulverized, crossed 40～80 purposes sieve, obtain malt meal;
(2) in malt meal, add phosphate buffer solution, centrifugal behind placement 40～60min, get supernatant, obtain the malt amylase crude extract;
(3) the malt amylase crude extract is carried out the intermittent type ultrasonic irradiation, ultrasonic irradiation 3s stops 1s～9s, UW mutual radiation 10～20min, and said ultrasonic frequency is 20kHz, ultrasonic power 120～160W;
The pH=5.8 of said phosphoric acid buffer;
Temperature of reaction system is lower than 4 ℃ during said ultrasonic irradiation;
The mass volume ratio of said malt meal and phosphate buffer solution is 1: 15～1: 10g/ml.
2. method according to claim 1 is characterized in that, the device of said pulverizing is a kibbler.
3. method according to claim 1 is characterized in that, said centrifugal rotation speed is 1500～3000r/min, and the time is 10～15min.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010102299861A CN101892220B (en) | 2010-07-16 | 2010-07-16 | Method for improving activity of malt amylase |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010102299861A CN101892220B (en) | 2010-07-16 | 2010-07-16 | Method for improving activity of malt amylase |
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| CN101892220A CN101892220A (en) | 2010-11-24 |
| CN101892220B true CN101892220B (en) | 2012-05-09 |
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| CN2010102299861A Expired - Fee Related CN101892220B (en) | 2010-07-16 | 2010-07-16 | Method for improving activity of malt amylase |
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Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103266032B (en) * | 2013-05-13 | 2014-05-07 | 湖南鸿鹰生物科技有限公司 | Total-effect beer compound enzyme |
| CN103255011B (en) * | 2013-05-13 | 2014-05-07 | 湖南鸿鹰生物科技有限公司 | Beer complex enzyme capable of improving beer malt fragrance |
| CN105039451A (en) * | 2015-07-25 | 2015-11-11 | 陈从龙 | Enzyme capable of directly and chemically synthesizing glucose in starch inside and outside human bodies |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1970781A (en) * | 2006-12-07 | 2007-05-30 | 天津大学 | Method for saccharification of lignocellulose by ultrasonic synergistic catalysis of modified cellulose |
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2010
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1970781A (en) * | 2006-12-07 | 2007-05-30 | 天津大学 | Method for saccharification of lignocellulose by ultrasonic synergistic catalysis of modified cellulose |
Non-Patent Citations (2)
| Title |
|---|
| Yaldagard M等.Investigation of the Effects of Ultrasound on Extraction of alpha-Amylase from the Flour of Malted Barley.《JOURNAL OF THE AMERICAN SOCIETY OF BREWING CHEMISTS》.2009,第67卷(第3期),141-145. * |
| 陈小丽等.超声波对淀粉酶催化活性的影响.《华南农业大学学报(自然科学版)》.2005,第26卷(第01期),76-79. * |
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