CN109913434A - A kind of complex enzyme and its preparation method and application - Google Patents
A kind of complex enzyme and its preparation method and application Download PDFInfo
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- CN109913434A CN109913434A CN201910276960.3A CN201910276960A CN109913434A CN 109913434 A CN109913434 A CN 109913434A CN 201910276960 A CN201910276960 A CN 201910276960A CN 109913434 A CN109913434 A CN 109913434A
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- complex enzyme
- climbing groundsel
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Abstract
The embodiment of the invention discloses a kind of complex enzyme and its preparation method and application, which includes the component of following parts by weight: 12-18 parts of cellobiase, 4-8 parts of zytase, 8-12 parts of cellulase, 8-12 parts of pectase, 8-12 parts of glycerol, 3-7 parts of sodium chloride and 38-50 parts of water；The above-mentioned complex enzyme of the present invention can cellulose, hemicellulose, cell wall to climbing groundsel crack, promote the release of climbing groundsel effective component, and it is precipitated and shortens settling time by the hydrolysis to stickums such as xylan, beta glucan, pectin, reduction material viscosity, promotion effective component, effectively improve the yield of climbing groundsel effective component.
Description
Technical field
The present embodiments relate to enzyme preparation technical fields, and in particular to a kind of complex enzyme and its preparation method and application.
Background technique
Climbing groundsel also known as nine inner bright, Herba Senecionis Scandentis, HERBA SENECIONIS SCANDENTIS, bright stroke of eye, sweet vernal grass Broomjutre Sida Herb, Kowloon light, nine ridge light.For chrysanthemum
Section herbaceous plant, be born under hillside, sparse woods, Lin Bian, roadside, in limes marginis thick grass.Originate in Henan, Shaanxi, Jiangsu, Zhejiang, Guangxi,
Sichuan.Its is cold in nature, bitter, has clearing heat and detoxicating, improving eyesight, antipruritic and other effects.It is chiefly used in anemopyretic cold, red eye, swell pain, diarrhea dysentery
Disease, skin eczema boil.
Climbing groundsel is a herb material, and the extraction process of effective component in climbing groundsel is generally water decoction or ethyl alcohol leaching
It mentions, currently, it is lower to decoct the recovery rate extracted to climbing groundsel effective component by water；And pass through the method pair of alcohol steep
What climbing groundsel effective component was extracted receives to increase, but the larger higher cost of the chemical levels such as ethyl alcohol, and pacifies to the person
Complete and environment has some impact on.
Summary of the invention
For this purpose, the embodiment of the present invention provides a kind of complex enzyme, to solve to soak using water boiling method or ethyl alcohol in the prior art
Formulation carries out effective component in climbing groundsel, and to extract recovery rate lower, at high cost and the problem of impact to human body and environment.
To achieve the goals above, the embodiment of the present invention provides the following technical solutions:
A kind of complex enzyme is provided according to a first aspect of the embodiments of the present invention, and the complex enzyme includes the group of following parts by weight
Point:
12-18 parts of cellobiase, 4-8 parts of zytase, 8-12 parts of cellulase, 8-12 parts of pectase, glycerol 8-12
Part, 3-7 parts of sodium chloride and 38-50 parts of water.
The present invention, being capable of cellulose, hemicellulose, cell to climbing groundsel by the selection of above-mentioned specific components and dosage
Wall is cracked, and can promote the release of climbing groundsel effective component, and by goos such as xylan, beta glucan, pectin
The hydrolysis of matter reduces material viscosity, promotes effective component precipitation and shortens settling time, raising climbing groundsel effective component
Rate.
Further, the complex enzyme includes the component of following parts by weight: 14-16 parts of cellobiase, zytase 5-7
Part, 9-11 parts of cellulase, 9-11 parts of pectase, 9-11 parts of glycerol, 4-6 parts of sodium chloride and 40-46 parts of water；Preferably, described
Complex enzyme includes the component of following parts by weight: 15 parts of cellobiase, 6 parts of zytase, 10 parts of cellulase, pectase 10
Part, 10 parts of glycerol, 5 parts of sodium chloride and 44 parts of water.The present invention can be mentioned preferably by further limiting to dosage of each component
The yield of effective component in high climbing groundsel.
A kind of preparation method of complex enzyme is provided according to a second aspect of the embodiments of the present invention, and the preparation method includes such as
Lower step:
Each component is weighed according to the proportion, at room temperature, each component is mixed, is stirred up to the complex enzyme.
Further, the mixing speed is 200-300r/min；Mixing time is 25-35min.
A kind of application of complex enzyme in the extraction of climbing groundsel effective component is provided according to a third aspect of the embodiments of the present invention,
The complex enzyme is above-mentioned complex enzyme or the complex enzyme that above-mentioned preparation method is prepared.
A kind of extracting method of climbing groundsel effective component, the extracting method are provided according to a fourth aspect of the embodiments of the present invention
Include the following steps:
(a) climbing groundsel crushed, be placed in reaction kettle；
(b) complex enzyme that addition water and above-mentioned complex enzyme or above-mentioned preparation method are prepared in a kettle, and adjust
PH to 4.8-5.2, enzymatic hydrolysis；
(c) enzymolysis liquid is stood, supernatant is collected by filtration, by supernatant purification, be drying to obtain climbing groundsel effective component.
Further, the climbing groundsel effective component is general flavone.
Further, the smashed climbing groundsel mesh granularity is 10-20 mesh；The complex enzyme additive amount is a thousand li light quality
The 6-10 ‰ of amount；The water additive amount is 10-20 times of a thousand li light quality.
Further, the hydrolysis temperature is 45-55 DEG C；Time is 48-54h.
Further, the time of repose is 1-2h；The purification is filtered to filter supernatant by plate compression
Liquid is freeze-dried to obtain freeze-dried powder；The drying temperature is 35-45 DEG C.
The present invention is first crushed climbing groundsel by above-mentioned specific extracting method, can promote the analysis of effective component
Out, by the restriction of enzymatic hydrolysis condition, the activity of complex enzyme can be preferably improved, conducive to obtaining for climbing groundsel effective component is improved
Rate.
The embodiment of the present invention has the advantages that
(1) the above-mentioned complex enzyme of the present invention can cellulose, hemicellulose, cell wall to climbing groundsel crack, promote
The release of climbing groundsel effective component, and it is viscous by the hydrolysis to stickums such as xylan, beta glucan, pectin, reduction material
Degree promotes effective component to be precipitated and shorten settling time, effectively improves the yield of climbing groundsel effective component.
(2) said extracted method of the present invention can preferably improve the activity of complex enzyme, and promote the precipitation of effective component,
Improve the yield of climbing groundsel effective component.
Specific embodiment
Embodiments of the present invention are illustrated by particular specific embodiment below, those skilled in the art can be by this explanation
Content disclosed by book is understood other advantages and efficacy of the present invention easily, it is clear that described embodiment is the present invention one
Section Example, instead of all the embodiments.Based on the embodiments of the present invention, those of ordinary skill in the art are not doing
Every other embodiment obtained under the premise of creative work out, shall fall within the protection scope of the present invention.
Embodiment 1
One, complex enzyme
Above-mentioned complex enzyme includes the component of following parts by weight:
14 parts of cellobiase, 7 parts of zytase, 9 parts of cellulase, 11 parts of pectase, 11 parts of glycerol, 6 parts of sodium chloride
With 40 parts of water.
Two, preparation method
Above-mentioned complex enzyme is prepared via a method which to obtain:
Each component is weighed, at room temperature, each component is mixed, stirs 25min with 300r/min up to described compound
Enzyme.
Embodiment 2
One, complex enzyme
Above-mentioned complex enzyme includes the component of following parts by weight:
16 parts of cellobiase, 5 parts of zytase, cellulase 11 part, 9 parts of pectase, 9 parts of glycerol, 4 parts of sodium chloride and
46 parts of water.
Two, preparation method
Above-mentioned complex enzyme is prepared via a method which to obtain:
Each component is weighed, at room temperature, each component is mixed, stirs 35min with 200r/min up to described compound
Enzyme.
Embodiment 3
One, complex enzyme
Above-mentioned complex enzyme includes the component of following parts by weight:
15 parts of cellobiase, 6 parts of zytase, 10 parts of cellulase, 10 parts of pectase, 10 parts of glycerol, 5 parts of sodium chloride
With 44 parts of water.
Two, preparation method
Above-mentioned complex enzyme is prepared via a method which to obtain:
Each component is weighed, at room temperature, each component is mixed, stirs 30min with 250r/min up to described compound
Enzyme.
Embodiment 4
The present embodiment is a kind of extracting method of climbing groundsel effective component, which includes the following steps:
(a) climbing groundsel is crushed to 10-20 mesh, be then placed in reaction kettle；
(b) complex enzyme of the water of a thousand li light quality ... and the embodiment 3 of a thousand li light quality 8 ‰ is added in a kettle, and
Using salt acid for adjusting pH to 5.0,50h is digested at 50 DEG C；
(c) enzymolysis liquid is stood into 1.5min, supernatant is collected by filtration, supernatant is filtered by plate compression, is filtered
Liquid is freeze-dried to obtain freeze-dried powder, and climbing groundsel effective component is drying to obtain at 40 DEG C.
Qualitative detection is carried out to obtained effective component using circular paper chromatography method, testing result is that the effective component is total
Flavones.
Embodiment 5
The present embodiment is a kind of extracting method of climbing groundsel effective component, which includes the following steps:
(a) climbing groundsel is crushed to 10-20 mesh, be then placed in reaction kettle；
(b) complex enzyme of the water of a thousand li light quality ... and the embodiment 3 of a thousand li light quality 9 ‰ is added in a kettle, and
Using salt acid for adjusting pH to 5.1,53h is digested at 49 DEG C；
(c) enzymolysis liquid is stood into 2min, supernatant is collected by filtration, supernatant is filtered by plate compression, obtains filtrate
It is freeze-dried to obtain freeze-dried powder, climbing groundsel effective component is drying to obtain at 45 DEG C.
Qualitative detection is carried out to obtained effective component using the method for embodiment 4, testing result is that the effective component is
General flavone.
Reference examples 1
This reference examples is a kind of complex enzyme, and the complex enzyme and the complex enzyme of embodiment 3 are essentially identical, and difference is only that will be wooden
Dextranase replaces with the cellulase of same number；The preparation method of above-mentioned complex enzyme is the same as the preparation method in embodiment 3.
Reference examples 2
This reference examples is a kind of complex enzyme, and the complex enzyme and the complex enzyme of embodiment 3 are essentially identical, and difference is only that fruit
Glue enzyme replaces with two pool enzyme of fiber of same number；The preparation method of above-mentioned complex enzyme is the same as the preparation method in embodiment 3.
Reference examples 3
This reference examples is a kind of extracting method of climbing groundsel effective component, the extracting method of the extracting method and embodiment 4
Essentially identical, difference, which is only that in the step (b), adjusts pH value to 4.5.
Reference examples 4
This reference examples is a kind of extracting method of climbing groundsel effective component, the extracting method of the extracting method and embodiment 4
Essentially identical, difference is only that the complex enzyme that the complex enzyme of embodiment 3 is replaced with to reference examples 1 in the step (b).
Reference examples 5
This reference examples is a kind of extracting method of climbing groundsel effective component, the extracting method of the extracting method and embodiment 4
Essentially identical, difference is only that the complex enzyme that the complex enzyme of embodiment 3 is replaced with to reference examples 2 in the step (b).
Experimental example
Climbing groundsel is chosen, is divided into 5 groups at random, wherein first group of extracting method according to embodiment 4 extracts；The
Two groups of extracting methods according to reference examples 3 extract；Third group is extracted according to the extracting method of reference examples 4；4th group
It is extracted according to the extracting method of reference examples 5；The climbing groundsel effective component that each group is extracted is collected respectively；
The yield of each substance in each group effective component is detected respectively using spectrophotometry, general flavone extracts yield
(%)=C × a × V/w × 100%, C is the general flavone (mg/ml) in extracting solution in formula, and a is extension rate, and V is extracting liq
Product (ml), W are sampling amount (mg)；Testing result is as shown in table 1:
Table 1
| Group | Yield (%) |
| First group | 8.72 |
| Second group | 6.81 |
| Third group | 5.46 |
| 4th group | 5.79 |
As shown in Table 1: the extracting method of the embodiment of the present invention 4 can significantly improve the general flavone in climbing groundsel, improve
The extraction yield of general flavone.
Although above having used general explanation and specific embodiment, the present invention is described in detail, at this
On the basis of invention, it can be made some modifications or improvements, this will be apparent to those skilled in the art.Therefore,
These modifications or improvements without departing from theon the basis of the spirit of the present invention are fallen within the scope of the claimed invention.
Claims (10)
1. a kind of complex enzyme, which is characterized in that the complex enzyme includes the component of following parts by weight:
12-18 parts of cellobiase, 4-8 parts of zytase, 8-12 parts of cellulase, 8-12 parts of pectase, 8-12 parts of glycerol, chlorine
Change sodium 3-7 parts and water 38-50 parts.
2. complex enzyme according to claim 1, which is characterized in that the complex enzyme includes the component of following parts by weight:
14-16 parts of cellobiase, 5-7 parts of zytase, 9-11 parts of cellulase, 9-11 parts of pectase, 9-11 parts of glycerol, chlorine
Change sodium 4-6 parts and water 40-46 parts.
3. complex enzyme according to claim 1, which is characterized in that the complex enzyme includes the component of following parts by weight:
15 parts of cellobiase, 6 parts of zytase, 10 parts of cellulase, 10 parts of pectase, 10 parts of glycerol, 5 parts of sodium chloride and water
44 parts.
4. the preparation method of any complex enzyme of claim 1-3, which comprises the steps of:
Each component is weighed according to the proportion, at room temperature, each component is mixed, is stirred up to the complex enzyme.
5. the preparation method according to claim 4, which is characterized in that the mixing speed is 200-300r/min；Stirring
Time is 25-35min.
6. a kind of application of complex enzyme in the extraction of climbing groundsel effective component, which is characterized in that the complex enzyme is claim
The complex enzyme that any complex enzyme of 1-3 or preparation method described in claim 4 or 5 are prepared.
7. a kind of extracting method of climbing groundsel effective component, which comprises the steps of:
(a) climbing groundsel crushed, be placed in reaction kettle；
(b) water and any complex enzyme of claim 1-3 or preparation described in claim 4 or 5 are added in a kettle
The complex enzyme that method is prepared, and adjust pH to 4.8-5.2, enzymatic hydrolysis；
(c) enzymolysis liquid is stood, supernatant is collected by filtration, by supernatant purification, be drying to obtain climbing groundsel effective component.
8. extracting method according to claim 7, which is characterized in that the climbing groundsel effective component is general flavone.
9. extracting method according to claim 7 or 8, which is characterized in that the smashed climbing groundsel mesh granularity is 10-
20 mesh；The complex enzyme additive amount is the 6-10 ‰ of a thousand li light quality；The additive amount of the water is the 10-20 of a thousand li light quality
Times.
10. extracting method according to claim 7 or 8, which is characterized in that the hydrolysis temperature is 45-55 DEG C；Time is
48-54h。
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101857855A (en) * | 2010-07-05 | 2010-10-13 | 沅江浣溪沙酶技术有限公司 | Complex enzyme preparation and application in producing diosgenin |
| CN103695392A (en) * | 2013-12-16 | 2014-04-02 | 宁夏夏盛实业集团有限公司 | Pomace enzyme as well as preparation method and application thereof |
| CN107881161A (en) * | 2017-11-16 | 2018-04-06 | 宁夏夏盛实业集团有限公司 | The technique that a kind of multienzyme complex containing β glucuroides and its enzymolysis tealeaves improve tea perfume quality |
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2019
- 2019-04-08 CN CN201910276960.3A patent/CN109913434A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101857855A (en) * | 2010-07-05 | 2010-10-13 | 沅江浣溪沙酶技术有限公司 | Complex enzyme preparation and application in producing diosgenin |
| CN103695392A (en) * | 2013-12-16 | 2014-04-02 | 宁夏夏盛实业集团有限公司 | Pomace enzyme as well as preparation method and application thereof |
| CN107881161A (en) * | 2017-11-16 | 2018-04-06 | 宁夏夏盛实业集团有限公司 | The technique that a kind of multienzyme complex containing β glucuroides and its enzymolysis tealeaves improve tea perfume quality |
Non-Patent Citations (4)
| Title |
|---|
| 于敬等: ""生物酶解技术在中药提取中的应用"", 《现代药物与临床》 * |
| 曹长海等: ""提高木质纤维素酶解糖化效率的研究进展"", 《中国生物工程杂志》 * |
| 李筱玲等: ""千里光总黄酮提取方法研究"", 《陕西农业科学》 * |
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Application publication date: 20190621 |