CN105985941B - A kind of liquid alpha-amylase composite thermal stabilizer - Google Patents
A kind of liquid alpha-amylase composite thermal stabilizer Download PDFInfo
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- CN105985941B CN105985941B CN201510954515.XA CN201510954515A CN105985941B CN 105985941 B CN105985941 B CN 105985941B CN 201510954515 A CN201510954515 A CN 201510954515A CN 105985941 B CN105985941 B CN 105985941B
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- amylase
- liquid alpha
- alpha
- stabilizer
- composite thermal
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/96—Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2408—Glucanases acting on alpha -1,4-glucosidic bonds
- C12N9/2411—Amylases
- C12N9/2414—Alpha-amylase (3.2.1.1.)
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01001—Alpha-amylase (3.2.1.1)
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- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
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- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
The present invention relates to a kind of liquid alpha-amylase composite thermal stabilizer, which is made of calcium chloride and pulullan polysaccharide mixture.Concentration of each raw material in liquid alpha-amylase are as follows: 4-8g/L CaCl2, 15-30g/L pulullan polysaccharide.The heat stabilizer convenient sources, it is cheap, and there is good biocompatibility, nontoxic, it not can cause environmental pollution；It maintains the enzyme activity height of liquid alpha-amylase and thermal stability is strong, be a kind of high-quality and efficient liquid alpha-amylase stabilizer.
Description
Technical field
The present invention relates to a kind of liquid alpha-amylase composite thermal stabilizers, belong to technical field of bioengineering.
Background technique
Alpha-amylase is a kind of endo-amylase, its random hydrolysis α-Isosorbide-5-Nitrae-glucoside bond, generates dextrin and reduction
Sugar.Currently, alpha-amylase is widely used to the every field such as weaving, light industry industry, food, papermaking, medicine, oil exploitation.
Alpha-amylase is as a kind of biocatalyst, because the disadvantages of hot conditions half-life is short, easy in inactivation seriously affects
Its using effect.As made wine in the processing of the starch of textile industry, starch is just to start to dissolve under the high temperature conditions, so big
Alpha-amylase heat-resisting enough is just needed in most cases, normal temperature type enzyme is inactivated due to being very easy under the high temperature conditions, so
Thermal stability just becomes one of the factor for limiting its application.
Currently, the method for improving enzyme heat stability has: addition stabilizer, chemical modification, immobilization, protein engineering
Deng wherein it is a kind of most economical effective method that heat stabilizer is added in liquid enzyme formulation.
Summary of the invention
Object of the present invention is to overcome the deficiencies of the prior art and provide a kind of high-quality, efficient liquid alpha-amylase stabilizer.
The purpose of the present invention is what is realized using following scheme:
A kind of liquid alpha-amylase composite thermal stabilizer, it is characterised in that: each constituent component is in liquid alpha-amylase
Concentration is 4-8g/L CaCl2, 15-30g/L pulullan polysaccharide.
The invention has the following advantages that
(1) the heat stabilizer convenient sources, it is cheap.
(2) heat stabilizer has good biocompatibility, nontoxic, not can cause environmental pollution.
(3) constituent of the present invention is simple, and preparation method is easy, low in cost, maintain liquid alpha-amylase enzyme activity high and
Stability is strong, is a kind of high-quality and efficient liquid alpha-amylase stabilizer, to high activity, high stability alpha-amylase liquid preparation
Industrialized development be of great significance.
Specific embodiment
Below by specific embodiment, the present invention will be further elaborated, it should be appreciated that, following the description is only
In order to explain the present invention, and it is not limited thereof.
Embodiment 1
Each constituent component is added in liquid alpha-amylase, sufficiently dissolves, makes their ultimate density are as follows: 4g/L
CaCl2, 30g/L pulullan polysaccharide.
The quality condition of alpha-amylase composite thermal stabilizer of the invention is evaluated, the method is as follows:
It takes in 500mL distilled water, is placed in water bath with thermostatic control to be measured after being heated to 90 DEG C, 20mL is added in the aqueous solution
1g/L hay bacillus mesophilicα-diastase liquid, is then added CaCl2And pulullan polysaccharide, making its ultimate density is respectively 4g/L
And 30g/L.Start timing, 1ml sample is taken to be added in 1mL 2g/L starch solution at regular intervals, measures alphalise starch at this time
The enzyme activity of enzyme.Under the conditions of 1 enzyme-activity unit is defined as 40 DEG C in 1min 1mg enzyme hydrolysis amount of starch (mg).
Under the above conditions, under the conditions of test hay bacillus alpha-amylase adds calcium chloride solution in not heat stabilizers and only
Inactivation half-life period t1/2(time that relative surplus enzyme activity is 50%), and compared with the stablizing effect of composite thermal stabilizer work one,
It the results are shown in Table 1.
Table 1
| Group | Stabilizer is not added | Only plus calcium chloride | Add composite thermal stabilizer |
| Half-life period t1/2 (min) | 0.4 | 1.6 | 5.8 |
From table 1 it follows that only add calcium chloride to play stablizing effect improves to a certain extent, and compound thermostabilization is added
After agent, effect is more preferable, and inactivation half-life period obviously increases.
Embodiment 2
Each constituent component is added in liquid alpha-amylase, sufficiently dissolves, makes their ultimate density 8g/L
CaCl2, 15g/L pulullan polysaccharide.
Claims (1)
1. a kind of liquid alpha-amylase composite thermal stabilizer, it is characterised in that: each constituent component is dense in liquid alpha-amylase
Degree is 4-8g/L CaCl2, 15-30g/L pulullan polysaccharide.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510954515.XA CN105985941B (en) | 2015-12-20 | 2015-12-20 | A kind of liquid alpha-amylase composite thermal stabilizer |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510954515.XA CN105985941B (en) | 2015-12-20 | 2015-12-20 | A kind of liquid alpha-amylase composite thermal stabilizer |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105985941A CN105985941A (en) | 2016-10-05 |
| CN105985941B true CN105985941B (en) | 2019-04-05 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201510954515.XA Active CN105985941B (en) | 2015-12-20 | 2015-12-20 | A kind of liquid alpha-amylase composite thermal stabilizer |
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| Country | Link |
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| CN (1) | CN105985941B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107760664B (en) * | 2017-11-02 | 2020-06-09 | 江南大学 | Method for improving heat stability of linear chain malto-oligosaccharide generating enzyme |
| CN111019929A (en) * | 2019-12-24 | 2020-04-17 | 福州三合元生物科技有限公司 | Long-acting biological enzyme heat stabilizer and micro-channel continuous synthesis method thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1043527A (en) * | 1990-01-11 | 1990-07-04 | 李春华 | A kind of preparation method of starch syrup |
| CN1793384A (en) * | 2005-12-30 | 2006-06-28 | 韩景银 | Tech. for producing fluid syrup |
| CN101157912A (en) * | 2007-09-17 | 2008-04-09 | 东华大学 | Novel alpha-amylase heat stabilizer |
| CN101167721A (en) * | 2006-10-26 | 2008-04-30 | 凌沛学 | Compound preparation containing orlistat |
| CN101679987A (en) * | 2007-03-09 | 2010-03-24 | 丹尼斯科美国公司 | Alkaliphilic bacillus species alpha-amylase variants, compositions comprising alpha-amylase variants, and methods of use |
-
2015
- 2015-12-20 CN CN201510954515.XA patent/CN105985941B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1043527A (en) * | 1990-01-11 | 1990-07-04 | 李春华 | A kind of preparation method of starch syrup |
| CN1793384A (en) * | 2005-12-30 | 2006-06-28 | 韩景银 | Tech. for producing fluid syrup |
| CN101167721A (en) * | 2006-10-26 | 2008-04-30 | 凌沛学 | Compound preparation containing orlistat |
| CN101679987A (en) * | 2007-03-09 | 2010-03-24 | 丹尼斯科美国公司 | Alkaliphilic bacillus species alpha-amylase variants, compositions comprising alpha-amylase variants, and methods of use |
| CN101157912A (en) * | 2007-09-17 | 2008-04-09 | 东华大学 | Novel alpha-amylase heat stabilizer |
Non-Patent Citations (2)
| Title |
|---|
| 硼酸盐对细菌α-淀粉酶的稳定作用;李庆宇等;《微生物学通报》;19810630;摘要 |
| 黄原胶对α-淀粉酶耐热性及其在热变性过程中构象变化的影响;薛正莲等;《生物学杂志》;20000630;第17卷(第3期);摘要，结果与讨论部分 |
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| CN105985941A (en) | 2016-10-05 |
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Effective date of registration: 20191226 Address after: 350012 unit 2, 4-2-1, building 2, sogao Plaza, 245 Xiushan Road, Xindian Town, Jin'an District, Fuzhou City, Fujian Province Patentee after: Fuzhou Sanheyuan Biotechnology Co., Ltd Address before: Fengming Road District of Ji'nan City, Shandong province 250101 port development Shandong Jianzhu University Patentee before: shandong jianzhu university |
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Denomination of invention: A compound heat stabilizer for liquid a - amylase Effective date of registration: 20201217 Granted publication date: 20190405 Pledgee: Bank of Communications Ltd. Fujian branch Pledgor: Fuzhou Sanheyuan Biotechnology Co.,Ltd. Registration number: Y2020350000144 |