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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Abstract
The invention discloses a method for improving activity of malt amylase, which comprises the following steps of: crushing dried barley malt by using a crusher, adding phosphate buffer with the pH value of 5.8 into the malt powder, standing at room temperature for 40 to 60 minutes, and centrifuging to obtain a malt amylase crude extract; and radiating the malt amylase crude extract by using ultrasonic waves, wherein the ultrasonic conditions are that: the fixed ultrasonic frequency is 20kHz, the ultrasonic power is 120 to 160W, the total time of ultrasonic irradiation is 10 to 20 minutes, the ultrasonic irradiation is carried out for 3 seconds and then stopped for 1 to 9 seconds, and the temperature of the ultrasonic system is less than 4 DEG C. The method can obviously improve the activity of the malt amylase by 20 to 30 percent so as to improve the hydrolysis rate of starch, effectively reduce the production cost of brewing beer, save energy and resources, and achieve obvious 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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| CN2010102299861A CN101892220B (en) | 2010-07-16 | 2010-07-16 | Method for improving activity of malt amylase |
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| 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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| 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 |
| 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 |
| 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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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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