CN116262772A - Method for extracting cAMP in red jujube residues by water-alcohol double phase - Google Patents
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
- C07H1/08—Separation; Purification from natural products
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
- C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
- C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
- C07H19/16—Purine radicals
- C07H19/20—Purine radicals with the saccharide radical esterified by phosphoric or polyphosphoric acids
- C07H19/213—Purine radicals with the saccharide radical esterified by phosphoric or polyphosphoric acids containing cyclic phosphate
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
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Abstract
The invention belongs to the technical field of food deep processing, and discloses a method for extracting cAMP in red jujube residues by water-alcohol double phases, which comprises the steps of adding red jujube residue powder into an enzymolysis tank, mixing and stirring with softened water, and adding compound enzyme for enzymolysis; extracting the mixture after enzymolysis by NaOH, and taking supernatant; adding the supernatant into a water-alcohol biphasic system for shake extraction until the phase formation in the water-alcohol biphasic system is complete; nanofiltration is carried out on the water-alcohol biphasic system with complete phase formation by using a nanofiltration membrane, and a trapped product containing cAMP is obtained; concentrating to obtain trapped product cAMP, crystallizing the concentrated solution by ethanol, and purifying to obtain cAMP finished product. The method for extracting cAMP in the red jujube residues by using water-alcohol two phases has the advantages of simple operation, reduced extraction period, suitability for industrial production, no need of elution by using formic acid eluent, raw material consumption saving, cost reduction and reduced damage to cAMP molecular structure.
Description
Technical Field
The invention belongs to the technical field of food deep processing, and particularly relates to a method for extracting cAMP in red date residues by water-alcohol double phases.
Background
At present, the cultivation area of the jujube tree in China exceeds 1000 ten thousand mu, and the jujube tree accounts for 99% of the total world cultivation area. Fructus Jujubae belongs to fruit used as both medicine and food, and contains abundant Cyclic Adenosine Monophosphate (CAMP). Adenosine cyclophosphate (cAMP) is an important substance with physiological activity widely existing in organisms, has regulation and control effects on protein biosynthesis, can be widely involved in regulation of cell functions, can relax smooth muscle, dilate blood vessels, improve liver functions, promote nerve regeneration, inhibit division of cells on the outer layer of skin and convert functions of abnormal cells, promote activity of respiratory chain oxidase, improve myocardial hypoxia and the like, has been widely studied abroad on physiological activity and action mechanism, and relates to aspects of disease treatment, signal expression, gene replication and the like.
In the prior art, research on extracting cyclic adenosine monophosphate (cAMP) from jujube has been reported. The existing extraction method needs to carry out column chromatography for multiple times, has complicated process and longer extraction period, and is not beneficial to industrial production. The existing cAMP extraction mode utilizes formic acid eluent to perform elution, so that the problem of incomplete elution caused by insufficient formic acid eluent is easily caused.
Through the above analysis, the problems and defects existing in the prior art are as follows:
(1) The existing method for extracting cAMP from jujubes has complex process and longer extraction period, and is not beneficial to industrial production.
(2) The existing cAMP extraction mode utilizes formic acid eluent to perform elution, so that the problem of incomplete elution caused by insufficient formic acid eluent is easily caused.
Disclosure of Invention
Aiming at the problems existing in the prior art, the invention provides a method for extracting cAMP from red date residues by water-alcohol double phase.
The invention is realized in such a way that the method for extracting cAMP from the red date slag by water-alcohol double phase comprises the following steps:
step one, drying, crushing and sieving red date residues to obtain red date residue powder;
step two, adding the red jujube residue powder into an enzymolysis tank, mixing and stirring with softened water, and adding complex enzyme for enzymolysis;
step three, extracting the mixture subjected to enzymolysis in the step two on a shaking table according to a feed liquid ratio of 1:10-20 by using NaOH, and taking supernatant;
step four, adding the supernatant into a water-alcohol biphasic system for shake extraction until the phase formation in the water-alcohol biphasic system is complete;
step five, nanofiltration is carried out on the water-alcohol biphasic system with complete phase formation by using a nanofiltration membrane, and a trapped product containing cyclic adenosine monophosphate (cAMP) is obtained;
and step six, concentrating to obtain trapped product cyclic adenosine monophosphate (cAMP), crystallizing the concentrated solution by ethanol, and purifying to obtain a finished product of cyclic adenosine monophosphate (cAMP).
In the first step, the red date residues are dried at the temperature of 30-38 ℃ and are sieved by a sieve with 80-100 meshes.
In the second step, when the complex enzyme is added for enzymolysis, the added complex enzyme accounts for 0.08 to 0.15 percent of the weight of the red date slag powder, and the enzymolysis is carried out for 60 to 90 minutes under the condition of 40 to 55 ℃.
In the fourth step, when the supernatant is added into the water-alcohol biphasic system for vibration extraction, the addition amount of the supernatant accounts for 3-6% of the weight of the water-alcohol biphasic system, and the vibration extraction time is 30-90min.
In the fourth step, the weight fraction of ethanol in the water-ethanol biphasic system is 15-21%, and the temperature is 25 ℃.
In the sixth step, the concentration mode is reduced pressure concentration, the vacuum degree of reduced pressure concentration is 0.1-0.3 MPa, and the temperature is 30-45 ℃.
In the second step, the complex enzyme is a mixture of pectase, cellulase and flavor enzyme, and the preparation method of the complex enzyme comprises the following steps:
(1) Respectively weighing pectase, cellulase and flavourzyme with the weight ratio of 1:13-21:3-7;
(2) And adding the weighed pectase, cellulase and flavor enzyme into water under a stirring state, wherein the weight of the water is 14-18 times of the total weight of the pectase, the cellulase and the flavor enzyme, and obtaining the compound enzyme solution.
In the step (2), when the weighed pectase, cellulase and flavourzyme are added into water under stirring, the stirring speed is 180-300 r/min, and the water temperature is 30-40 ℃.
Further, the preparation method of the flavor enzyme comprises the following steps:
1) Fresh bean pulp and wheat bran are selected as raw materials, and dust removal and impurity removal are carried out;
2) Crushing soybean meal and wheat bran, sterilizing, adding lactobacillus and saccharomycetes, and fermenting for 2-4 days at normal temperature;
3) Extracting, separating and ultrafiltering to obtain solution after fermentation;
4) And freeze-drying and crushing the solution to obtain the flavor enzyme.
In the step 2), the sterilization mode is that high-temperature steam at 115-125 ℃ is utilized for sterilization, the sterilization time is 20-30 min, and the temperature is cooled to the room temperature.
By combining all the technical schemes, the invention has the advantages and positive effects that:
the method for extracting cAMP in the red jujube residues by using water-alcohol two phases has the advantages of simple operation, reduced extraction period, suitability for industrial production, no need of elution by using formic acid eluent, raw material consumption saving, cost reduction and reduced damage to cAMP molecular structure. The invention adopts the enzyme method to reduce the material viscosity and combines the molecular nanofiltration membrane nanofiltration technology to extract the effective components in the red jujube residues, has the advantages of rich raw material sources, simple process, large treatment capacity, high extraction rate, high product purity and the like, belongs to natural and nontoxic products, and can be used as medical raw materials or food additives; the extracted byproduct red date juice can realize deep processing of red dates, promote an industrial chain of comprehensive development, realize pollution-free production, solve the problems of complicated process, long period, low product purity and the like in the prior art, reduce cost, provide a better way for deep processing of red date raw materials, and are more suitable for industrialized production and application.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present application, the following description will briefly explain the drawings needed in the embodiments of the present application, and it is obvious that the drawings described below are only some embodiments of the present application, and that other drawings can be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a flow chart of a method for extracting cAMP from red date slag by water-alcohol two-phase extraction, which is provided by the embodiment of the invention.
FIG. 2 is a flow chart of a preparation method of the complex enzyme provided by the embodiment of the invention.
FIG. 3 is a flowchart of a method for preparing a flavoring enzyme according to an embodiment of the present invention.
Detailed Description
The present invention will be described in further detail with reference to the following examples in order to make the objects, technical solutions and advantages of the present invention more apparent. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the invention.
Aiming at the problems existing in the prior art, the invention provides a method for extracting cAMP from red date residues by water-alcohol double phase, and the invention is described in detail below with reference to the accompanying drawings.
As shown in FIG. 1, the method for extracting cAMP from red date residues by water-alcohol biphasic extraction provided by the embodiment of the invention comprises the following steps:
s101, drying, crushing and sieving red date residues to obtain red date residue powder;
s102, adding red jujube residue powder into an enzymolysis tank, mixing and stirring with softened water, and adding complex enzyme for enzymolysis;
s103, extracting the mixture subjected to enzymolysis in the second step on a shaking table according to a feed liquid ratio of 1:10-20 by using NaOH, and taking supernatant;
s104, adding the supernatant into a water-alcohol biphasic system for shake extraction until the phase formation in the water-alcohol biphasic system is complete;
s105, nanofiltration is carried out on the water-alcohol biphasic system with complete phase formation by using a nanofiltration membrane, and a trapped product containing cAMP is obtained;
s106, concentrating to obtain trapped product cAMP, crystallizing and purifying the concentrated solution by ethanol to obtain a finished product of cAMP.
In the step S101 of the embodiment of the invention, the red date residues are dried at the temperature of 30-38 ℃ and are sieved by a sieve of 80-100 meshes.
In the step S102 of the embodiment of the invention, when the complex enzyme is added for enzymolysis, the added complex enzyme accounts for 0.08 to 0.15 percent of the weight of the red date slag powder, and the enzymolysis is carried out for 60 to 90 minutes under the condition of 40 to 55 ℃.
In step S104 of the embodiment of the invention, when the supernatant is added into the water-alcohol biphasic system for vibration extraction, the addition amount of the supernatant accounts for 3-6% of the weight of the water-alcohol biphasic system, and the vibration extraction time is 30-90min.
In step S104 of the embodiment of the invention, the weight fraction of ethanol in the water-ethanol biphasic system is 15-21%, and the temperature is 25 ℃.
In step S106 of the embodiment of the invention, the concentration mode is reduced pressure concentration, the vacuum degree of reduced pressure concentration is 0.1-0.3 MPa, and the temperature is 30-45 ℃.
The complex enzyme in the embodiment of the invention is a mixture of pectase, cellulase and flavourzyme.
As shown in FIG. 2, the preparation method of the complex enzyme in the embodiment of the invention comprises the following steps:
s201, respectively weighing pectase, cellulase and flavourzyme with the weight ratio of 1:13-21:3-7;
s202, adding the weighed pectase, cellulase and flavor enzyme into water under a stirring state, wherein the weight of the water is 14-18 times of the total weight of the pectase, the cellulase and the flavor enzyme, and obtaining the complex enzyme solution.
In step S202 of the embodiment of the invention, when the weighed pectase, cellulase and flavourzyme are added into water under stirring, the stirring speed is 180-300 r/min, and the water temperature is 30-40 ℃.
As shown in fig. 3, the preparation method of the flavourzyme in the embodiment of the present invention includes:
s301, selecting fresh bean pulp and wheat bran as raw materials, and removing dust and impurities;
s302, crushing soybean meal and wheat bran, sterilizing, adding lactobacillus and saccharomycetes, and fermenting for 2-4 days at normal temperature;
s303, extracting, separating and ultrafiltering after fermentation to obtain a solution;
s304, freeze-drying and crushing the solution to obtain the flavor enzyme.
In step S302 of the embodiment of the invention, the sterilization mode is that high-temperature steam at 115-125 ℃ is utilized for sterilization, the sterilization time is 20-30 min, and the sterilization is cooled to room temperature.
While the invention has been described with respect to what is presently considered to be the most practical and preferred embodiments, it is to be understood that the invention is not limited to the disclosed embodiments, but on the contrary, is intended to cover various modifications, equivalents, and alternatives falling within the spirit and scope of the invention.
Claims (10)
1. A method for extracting cAMP in red date slag by water-alcohol double phase, which is characterized in that the method for extracting cAMP in red date slag by water-alcohol double phase comprises the following steps:
step one, drying, crushing and sieving red date residues to obtain red date residue powder;
step two, adding the red jujube residue powder into an enzymolysis tank, mixing and stirring with softened water, and adding complex enzyme for enzymolysis;
step three, extracting the mixture subjected to enzymolysis in the step two on a shaking table according to a feed liquid ratio of 1:10-20 by using NaOH, and taking supernatant;
step four, adding the supernatant into a water-alcohol biphasic system for shake extraction until the phase formation in the water-alcohol biphasic system is complete;
step five, nanofiltration is carried out on the water-alcohol biphasic system with complete phase formation by using a nanofiltration membrane, and a trapped product containing cAMP is obtained;
step six, concentrating to obtain trapped product cAMP, crystallizing and purifying the concentrated solution by ethanol to obtain a finished product of cAMP.
2. The method for extracting cAMP from red date slag by water-alcohol double phase according to claim 1, wherein in the first step, the red date slag is dried at the temperature of 30-38 ℃ and is sieved by a sieve of 80-100 meshes.
3. The method for extracting cAMP from red date slag by water-alcohol double phase extraction as claimed in claim 1, wherein in the second step, when complex enzyme is added for enzymolysis, the added complex enzyme is 0.08-0.15% of the weight of red date slag powder, and the enzymolysis is carried out for 60-90 minutes under the condition of 40-55 ℃.
4. The method for extracting cAMP from red jujube residues by water-alcohol biphasic extraction as claimed in claim 1, wherein in the fourth step, when supernatant is added into a water-alcohol biphasic system for shake extraction, the addition amount of the supernatant accounts for 3-6% of the weight of the water-alcohol biphasic system, and the shake extraction time is 30-90min.
5. The method for extracting cAMP from red jujube residue by water-alcohol biphasic extraction according to claim 1, wherein in the fourth step, the weight fraction of ethanol in the water-ethanol biphasic system is 15-21%, and the temperature is 25 ℃.
6. The method for extracting cAMP from red jujube residues by water-alcohol biphasic extraction according to claim 1, wherein in the sixth step, a concentration mode is adopted, wherein the concentration mode is reduced pressure concentration, the vacuum degree of reduced pressure concentration is 0.1-0.3 MPa, and the temperature is 30-45 ℃.
7. The method for extracting cAMP from red jujube residues by water-alcohol biphasic extraction according to claim 1, wherein in the second step, the complex enzyme is a mixture of pectase, cellulase and flavourzyme, and the preparation method of the complex enzyme comprises the following steps:
(1) Respectively weighing pectase, cellulase and flavourzyme with the weight ratio of 1:13-21:3-7;
(2) And adding the weighed pectase, cellulase and flavor enzyme into water under a stirring state, wherein the weight of the water is 14-18 times of the total weight of the pectase, the cellulase and the flavor enzyme, and obtaining the compound enzyme solution.
8. The method for extracting cAMP from red jujube residues by water-alcohol biphasic extraction according to claim 7, wherein in the step (2), when the weighed pectase, cellulase and flavourzyme are added into water under stirring, the stirring speed is 180-300 r/min, and the water temperature is 30-40 ℃.
9. The method for extracting cAMP from red date slag by water-alcohol double phase according to claim 7, wherein the method for preparing the flavor enzyme comprises the following steps:
1) Fresh bean pulp and wheat bran are selected as raw materials, and dust removal and impurity removal are carried out;
2) Crushing soybean meal and wheat bran, sterilizing, adding lactobacillus and saccharomycetes, and fermenting for 2-4 days at normal temperature;
3) Extracting, separating and ultrafiltering to obtain solution after fermentation;
4) And freeze-drying and crushing the solution to obtain the flavor enzyme.
10. The method for extracting cAMP from red jujube residues by water-alcohol biphasic extraction according to claim 9, wherein in the step 2), a sterilization mode is adopted, wherein the sterilization mode is that high-temperature steam at 115-125 ℃ is utilized for sterilization, the sterilization time is 20-30 min, and the cooling is carried out to room temperature.
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| EP2221058A1 (en) * | 2007-11-30 | 2010-08-25 | Chi, Yu-Fen | Antimelancholic medicine prepared with jujube camp materials |
| CN102382161A (en) * | 2011-08-26 | 2012-03-21 | 食品行业生产力促进中心 | Method for extracting red date cyclic adenosine monophosphate through ultrasonic composite membrane technology |
| CN109400663A (en) * | 2018-11-20 | 2019-03-01 | 天津核生科技生物工程有限公司 | The method for extracting cyclic adenosine monophosphate in golden jujube and the application on pre- anti-cancer |
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2021
- 2021-12-14 CN CN202111519800.0A patent/CN116262772A/en active Pending
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|---|---|---|---|---|
| CN1948329A (en) * | 2006-11-09 | 2007-04-18 | 常月梅 | Technology of extracting adenosin phosphoric acid from date |
| EP2221058A1 (en) * | 2007-11-30 | 2010-08-25 | Chi, Yu-Fen | Antimelancholic medicine prepared with jujube camp materials |
| CN101525363A (en) * | 2009-04-24 | 2009-09-09 | 天津科技大学 | Method for extracting cyclic adenosine monophosphate (cAMP) from red jujubes |
| CN102382161A (en) * | 2011-08-26 | 2012-03-21 | 食品行业生产力促进中心 | Method for extracting red date cyclic adenosine monophosphate through ultrasonic composite membrane technology |
| CN109400663A (en) * | 2018-11-20 | 2019-03-01 | 天津核生科技生物工程有限公司 | The method for extracting cyclic adenosine monophosphate in golden jujube and the application on pre- anti-cancer |
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