CN114107265A - Process for extracting bromelain - Google Patents
Process for extracting bromelain Download PDFInfo
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- CN114107265A CN114107265A CN202111308128.0A CN202111308128A CN114107265A CN 114107265 A CN114107265 A CN 114107265A CN 202111308128 A CN202111308128 A CN 202111308128A CN 114107265 A CN114107265 A CN 114107265A
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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)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
- C12N9/63—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from plants
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
- C12Y304/22—Cysteine endopeptidases (3.4.22)
- C12Y304/22033—Fruit bromelain (3.4.22.33), i.e. juice bromelain
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Abstract
The invention provides a process for extracting bromelain, and relates to the technical field of extraction of plant enzymes. A process for extracting bromelain, comprising the steps of: cleaning pineapple peel, crushing and squeezing to obtain pineapple peel juice; freezing pineapple peel juice, adding a coagulant, and precipitating and centrifuging to obtain clear juice; performing ultrafiltration treatment on the clear juice to obtain a bromelain concentrated solution; adding acid into the bromelain concentrated solution to adjust pH to acidity, cooling and standing to generate precipitate, centrifuging at high speed to obtain enzyme paste, and freeze drying the enzyme paste to obtain dried bromelain finished product. The method comprises the steps of crushing, squeezing, freezing, precipitating, centrifuging, ultrafiltering, high-speed centrifuging and the like, so that the bromelain is extracted, the steps are simple, and the extracted bromelain has the characteristics of high enzyme activity and less impurities.
Description
Technical Field
The invention relates to the technical field of extraction of plant enzymes, and particularly relates to a process for extracting bromelain.
Background
Bromelain (broomelain, EC3.4.22.33) is a proteolytic enzyme present in pineapple plants (including fruits, stems, stalks and leaves), and has multiple isoenzymes, and its active center contains a sulfhydryl group (-SH), so it belongs to a sulfhydryl protease. The bromelain is widely used in the industries of medicine, food, cosmetics, textile, feed, leather and the like. It can decompose protein such as fibrin and hemoglobin, dissolve fibrin and blood clot under the condition of small side effect, further improve body fluid circulation, increase tissue permeability, and cause inflammation and edema to subside; the composition can be used together with antibiotic and chemotherapy medicine to promote the penetration and diffusion of medicine to focus, thereby enhancing therapeutic effect.
The protein extraction and purification method mainly comprises salting out, isoelectric precipitation, organic solvent precipitation, tannin (tannic acid) precipitation, ion exchange chromatography, affinity chromatography, ultrafiltration concentration, etc. The production process of bromelain mainly comprises an ammonium sulfate precipitation method, a tannin precipitation method, an ultrafiltration method and the like, and the methods have the following defects: the ammonium sulfate precipitation method needs a large amount of ammonium sulfate, and the environmental pollution is serious; the enzyme preparation obtained by the tannin precipitation method is insoluble in water, and the enzyme activity is low; the ultrafiltration method requires large equipment investment, the viscosity is gradually increased in the ultrafiltration concentration process of the pineapple juice, and the ultrafiltration membrane is easy to block, so the production period is long and the extraction yield is low.
Disclosure of Invention
The invention aims to provide a process for extracting bromelain, which can effectively extract bromelain from pineapple peels.
The technical problem to be solved by the invention is realized by adopting the following technical scheme.
The embodiment of the application provides a process for extracting bromelain, which comprises the following steps:
cleaning pineapple peel, crushing and squeezing to obtain pineapple peel juice;
freezing pineapple peel juice, adding a coagulant, and precipitating to obtain clear juice;
performing ultrafiltration treatment on the clear juice to obtain a bromelain concentrated solution;
adding acid into the bromelain concentrated solution to adjust pH to acidity, cooling and standing to generate precipitate, centrifuging at high speed to obtain enzyme paste, and freeze drying the enzyme paste to obtain dried bromelain finished product.
Compared with the prior art, the embodiment of the invention has at least the following advantages or beneficial effects:
the method comprises the steps of crushing, squeezing, freezing, precipitating, primary ultrafiltration, secondary ultrafiltration, high-speed centrifugation and the like, so that the bromelain is obtained, the steps are simple, no preservative is added in the whole production process, so that the bromelain activity is not lost, the extracted bromelain has the characteristics of high enzyme activity, high purity and less impurities and is far higher than the industrial standard, and the pineapple juice permeate obtained after secondary ultrafiltration can be prepared into a high-concentration pineapple product after twice concentration, so that the pineapple peel can be comprehensively utilized.
The invention freezes the pressed pineapple peel juice, then adds the gelling agent to precipitate the pineapple peel juice at low temperature, can accelerate the aggregation of flocculates in the pineapple peel juice, can accelerate the precipitation speed by adopting a precipitation method of low-temperature standing, and improves the production efficiency of the pineapple enzymes.
The coagulant releasing agent is polyacrylamide and chitosan acetic acid solution with the molecular weight of 1800 ten thousand which are added in sequence, and the flocculation of floccules and impurities in the pineapple enzyme juice can be accelerated by matching the polyacrylamide and the chitosan acetic acid solution and adding the polyacrylamide and the chitosan acetic acid solution in a fixed sequence, so that the precipitation efficiency is improved.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.
FIG. 1 is a process flow diagram of an embodiment of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
It should be noted that the embodiments and features of the embodiments in the present application may be combined with each other without conflict. The present invention will be described in detail below with reference to specific examples.
A process for extracting bromelain, comprising the steps of:
cleaning pineapple peel, crushing and squeezing to obtain pineapple peel juice;
freezing pineapple peel juice, adding a coagulant, and precipitating and centrifuging to obtain clear juice;
performing ultrafiltration treatment on the clear juice to obtain a bromelain concentrated solution;
adding acid into the bromelain concentrated solution to adjust pH to acidity, cooling and standing to generate precipitate, centrifuging at high speed to obtain enzyme paste, and freeze drying the enzyme paste to obtain dried bromelain finished product.
In some embodiments of the present invention, if the requirement on the product quality is higher, the enzyme paste may be dissolved in 50 ° ethanol, the precipitation is performed at a high speed at 10 ℃, and the enzyme paste after the precipitation is separated is then vacuum freeze-dried to obtain a bromelain product with higher purity.
In some embodiments of the present invention, the pineapple peel is broken to 0.1-0.5 cm. The crushing of the pineapple peel can reduce the time for subsequent pressing, and multiple experiments show that the crushing of the pineapple peel to the granularity can produce more liquid compared with the crushing of the uncrushed pineapple peel under the condition of the same time and pressing force.
In some embodiments of the present invention, the squeezed pineapple peel juice is filtered through a 40-70 mesh sieve. The squeezed pineapple juice is filtered and then precipitated, so that larger impurities in the pineapple juice can be removed, and the subsequent estimated use amount is reduced.
In some embodiments of the present invention, the freezing temperature is 2 to 8 ℃ and the freezing time is 3 to 12 hours. The pineapple enzyme juice is frozen and then filtered, so that the precipitation effect of the pineapple enzyme can be improved, and the fact that the freezing low temperature has a promoting effect on the pineapple enzyme is shown.
In some embodiments of the present invention, the above-mentioned gelling agent is composed of a component a and a component B, wherein the component a is polyacrylamide with a molecular weight of 1800 ten thousand, and the ratio of the added amount to the weight of the pineapple peel juice is (1-1.4): 10000; the component B is chitosan liquid, and the mass ratio of chitosan to pineapple peel juice is (1-1.4): 1000. the step of adding the coagulant release agent is to add the component A, add the component B after stirring uniformly, and then stir uniformly, and the precipitation can be completed 3 hours after the coagulant release agent is added by adopting the method, and the supernatant is extracted.
In some embodiments of the present invention, the chitosan solution is prepared by dissolving chitosan with molecular weight of 2000 and high viscosity in diluted acetic acid.
In some embodiments of the present invention, the ultrafiltration treatment includes a primary ultrafiltration treatment and a secondary ultrafiltration treatment, the pore size of the ultrafiltration membrane of the primary ultrafiltration treatment is 180000 to 200000 molecular weight, and the pore size of the ultrafiltration membrane of the secondary ultrafiltration treatment is 5000 to 6000 molecular weight. The bromelain can pass through the filter membrane with the molecular weight of 180000-200000 but can not pass through the filter membrane with the molecular weight of 5000-6000; by adopting different ultrafiltration membranes, various components in the pineapple peel juice can be separated.
In some embodiments of the present invention, the acid is hydrochloric acid or acetic acid, and the adjusted pH is 3 to 5. The acidic environment is more favorable for precipitation.
In some embodiments of the present invention, the cooling temperature is 2 to 4 ℃ and the standing time is 1 to 2 hours.
In some embodiments of the present invention, the rotation speed of the high-speed centrifugation is 13000-18000 rpm.
The features and properties of the present invention are described in further detail below with reference to examples.
Example 1
A process for extracting bromelain, comprising the steps of:
adding water into fresh pineapple peels, cleaning the fresh pineapple peels in a peel washer, treating the cleaned sewage through sewage equipment, performing agricultural irrigation, conveying the cleaned pineapple peels to a crushing device through a peel loosening machine, crushing the pineapple peels into 0.1-0.5 cm-sized crushed residues, conveying the crushed pineapple peel residues to a squeezer, squeezing the crushed pineapple peel residues through a 50-mesh filter screen to obtain pineapple peel juice and pineapple peel residues, and performing silage fermentation on the pineapple peel residues to obtain silage;
freezing pineapple peel juice in freezing device at 6 deg.C for 8 hr, taking out, adding polyacrylamide with molecular weight of 1800 ten thousand, and adding amount of pineapple peel juice
Stirring, adding high viscosity chitosan acetic acid solution with molecular weight of 2000 with addition amount of 1.2 ‰, mixing, standing for precipitation, and sucking out supernatant via siphon device;
subjecting the clear juice to primary ultrafiltration treatment with an ultrafiltration membrane aperture of 200000 molecular weight, then subjecting the macromolecule concentrated solution to secondary ultrafiltration treatment with an ultrafiltration membrane aperture of 6000 molecular weight to obtain upper-layer bromelain concentrated solution and micromolecule permeate, subjecting the micromolecule permeate to reverse osmosis concentration and single-effect concentration to obtain pineapple concentrated solution, and sequentially obtaining pineapple concentrated solution with higher refractive index;
adding hydrochloric acid into the bromelain concentrated solution to adjust pH to 3.5, cooling at 3 deg.C, standing for 1.5 hr to produce precipitate, centrifuging at high speed of 15000 r/min to obtain enzyme paste, and freeze drying to obtain dried bromelain product.
Example 2
A process for extracting bromelain, comprising the steps of:
adding water into fresh pineapple peels, cleaning the fresh pineapple peels in a peel washer, treating the cleaned sewage through sewage equipment, performing agricultural irrigation, conveying the cleaned pineapple peels to a crushing device through a peel loosening machine, crushing the pineapple peels into crushed slag with the size of 0.1-0.5 cm, conveying the crushed slag of the pineapple peels to a squeezer, squeezing the crushed slag of the pineapple peels, filtering the crushed slag of the pineapple peels through a 60-mesh filter screen to obtain pineapple peel juice and pineapple peel slag, and performing silage fermentation on the pineapple peel slag to obtain silage;
freezing pineapple peel juice in a freezing device at 8 deg.C for 12 hr, taking out, adding polyacrylamide with molecular weight of 1800 ten thousand, and adding into pineapple peel juice
Stirring, adding high viscosity chitosan solution with molecular weight of 2000, adding 1.4 ‰ of pineapple peel juice, mixing, standing for precipitation, sucking out supernatant via siphon device to obtain green juice, discharging residue, and processing residue to obtain feed;
and (2) subjecting the clear juice to primary ultrafiltration treatment with an ultrafiltration membrane pore diameter of 200000 molecular weight, then subjecting the macromolecular concentrated solution to secondary ultrafiltration treatment with an ultrafiltration membrane pore diameter of 6000 molecular weight to obtain upper-layer bromelain concentrated solution and micromolecular permeate, and subjecting the micromolecular permeate to reverse osmosis concentration and single-effect concentration to obtain pineapple concentrated solution, and sequentially obtaining the pineapple concentrated solution with higher refractive index.
Adding hydrochloric acid into the bromelain concentrated solution to adjust pH to 4, cooling at 3 deg.C, standing for 2 hr to produce precipitate, centrifuging at high speed of 16000 rpm to obtain enzyme paste, and freeze drying to obtain dried bromelain product.
Example 3
A process for extracting bromelain, comprising the steps of:
adding water into fresh pineapple peels, cleaning the fresh pineapple peels in a peel washer, treating the cleaned sewage through sewage equipment, performing agricultural irrigation, conveying the cleaned pineapple peels to a crushing device through a peel loosening machine, crushing the pineapple peels into 0.1-0.5 cm-sized crushed residues, conveying the crushed pineapple peel residues to a squeezer, squeezing the crushed pineapple peel residues through a 40-mesh filter screen to obtain pineapple peel juice and pineapple peel residues, and performing silage fermentation on the pineapple peel residues to obtain silage;
freezing pineapple peel juice in a freezing device at 2 deg.C for 3 hr, taking out, adding polyacrylamide with molecular weight of 1800 ten thousand, and adding into pineapple peel juice
Stirring, adding high viscosity chitosan solution with molecular weight of 2000, adding 1.2 ‰ of pineapple peel juice, mixing, standing for precipitation, sucking out supernatant via siphon device to obtain green juice, discharging residue, and processing residue to obtain feed;
performing primary ultrafiltration treatment on the clear juice with the aperture of 180000 molecular weight by using an ultrafiltration membrane, performing secondary ultrafiltration treatment on the macromolecular concentrated solution with the aperture of 5000 molecular weight by using the ultrafiltration membrane to obtain upper-layer bromelain concentrated solution and micromolecular permeate, performing reverse osmosis concentration and single-effect concentration on the micromolecular permeate to obtain pineapple concentrated solution, and sequentially obtaining pineapple concentrated solution with higher refractive index;
adding hydrochloric acid into the bromelain concentrated solution to adjust pH to 5, cooling at 4 deg.C, standing for 1 hr to produce precipitate, centrifuging at 18000 rpm to obtain enzyme paste, and freeze drying to obtain dried bromelain product.
Example 4
A process for extracting bromelain, comprising the steps of:
adding water into fresh pineapple peels, cleaning the fresh pineapple peels in a peel washer, treating the cleaned sewage through sewage equipment, performing agricultural irrigation, conveying the cleaned pineapple peels to a crushing device through a peel loosening machine, crushing the pineapple peels into 0.1-0.5 cm-sized crushed residues, conveying the crushed pineapple peel residues to a squeezer, squeezing the crushed pineapple peel residues through a 70-mesh filter screen to obtain pineapple peel juice and pineapple peel residues, and performing silage fermentation on the pineapple peel residues to obtain silage;
freezing pineapple peel juice in a freezing device at 6 deg.C for 10 hr, taking out, adding polyacrylamide with molecular weight of 1800 ten thousand, wherein the addition amount is pineapple peel juice
Stirring, adding high viscosity chitosan solution with molecular weight of 2000 (1.2 ‰) of pineapple peel juice, mixing, standing for precipitation, siphoning out supernatant to obtain green juice, discharging residue, and processing residue to obtain feed;
performing primary ultrafiltration treatment on the clear juice with the aperture of 180000 molecular weight by using an ultrafiltration membrane, performing secondary ultrafiltration treatment on the macromolecular concentrated solution with the aperture of 5500 molecular weight by using the ultrafiltration membrane to obtain upper-layer bromelain concentrated solution and micromolecular permeate, performing reverse osmosis concentration and single-effect concentration on the micromolecular permeate to obtain pineapple concentrated solution, and sequentially obtaining the pineapple concentrated solution with higher refractive index;
adding hydrochloric acid into the bromelain concentrated solution to adjust pH to 4.5, cooling at 2 deg.C, standing for 1.5 hr to produce precipitate, centrifuging at 18000 rpm to obtain enzyme paste, and freeze drying to obtain dried bromelain product.
Example 5
A process for extracting bromelain, comprising the steps of:
adding water into fresh pineapple peels, cleaning the fresh pineapple peels in a peel washer, treating the cleaned sewage through sewage equipment, performing agricultural irrigation, conveying the cleaned pineapple peels to a crushing device through a peel loosening machine, crushing the pineapple peels into crushed slag with the size of 0.1-0.5 cm, conveying the crushed slag of the pineapple peels to a squeezer, squeezing the crushed slag of the pineapple peels, filtering the crushed slag of the pineapple peels through a 60-mesh filter screen to obtain pineapple peel juice and pineapple peel slag, and performing silage fermentation on the pineapple peel slag to obtain silage;
freezing pineapple peel juice in a freezing device at 7 deg.C for 4 hr, taking out, adding polyacrylamide with molecular weight of 1800 ten thousand, wherein the addition amount is pineapple peel juice
Stirring, adding high viscosity chitosan solution with molecular weight of 2000 (1.2 ‰) of pineapple peel juice, mixing, standing for precipitation, siphoning out supernatant to obtain green juice, discharging residue, and processing residue to obtain feed;
performing primary ultrafiltration treatment on the clear juice with the aperture of 200000 molecular weight through an ultrafiltration membrane, performing secondary ultrafiltration treatment on the macromolecular concentrated solution with the aperture of 5000 molecular weight through the ultrafiltration membrane to obtain upper-layer bromelain concentrated solution and micromolecule permeate, performing reverse osmosis concentration and single-effect concentration on the micromolecule permeate to obtain pineapple concentrated solution, and sequentially obtaining pineapple concentrated solution with higher refractive index;
adding hydrochloric acid into the bromelain concentrated solution to adjust pH to 4.7, cooling at 2 deg.C, standing for 2 hr to produce precipitate, centrifuging at 18000 rpm to obtain enzyme paste, and freeze drying to obtain dried bromelain product.
Example 6
A process for extracting bromelain, comprising the steps of:
adding water into fresh pineapple peels, cleaning the fresh pineapple peels in a peel washer, treating the cleaned sewage through sewage equipment, performing agricultural irrigation, conveying the cleaned pineapple peels to a crushing device through a peel loosening machine, crushing the pineapple peels into 0.1-0.5 cm-sized crushed residues, conveying the crushed pineapple peel residues to a squeezer, squeezing the crushed pineapple peel residues through a 40-mesh filter screen to obtain pineapple peel juice and pineapple peel residues, and performing silage fermentation on the pineapple peel residues to obtain silage;
freezing pineapple peel juice in a freezing device at 6 deg.C for 10 hr, taking out, adding polyacrylamide with molecular weight of 1800 ten thousand, wherein the addition amount is pineapple peel juice
Stirring, adding high viscosity chitosan solution with molecular weight of 2000 (1.2 ‰) of pineapple peel juice, mixing, standing for precipitation, siphoning out supernatant to obtain green juice, discharging residue, and processing residue to obtain feed;
performing primary ultrafiltration treatment on the clear juice with the aperture of 160000 molecular weight by using an ultrafiltration membrane, performing secondary ultrafiltration treatment on the macromolecular concentrated solution with the aperture of 6000 molecular weight by using the ultrafiltration membrane to obtain upper-layer bromelain concentrated solution and micromolecular permeate, performing reverse osmosis concentration and single-effect concentration on the micromolecular permeate to obtain pineapple concentrated solution, and sequentially obtaining the pineapple concentrated solution with higher refractive index;
adding hydrochloric acid into the bromelain concentrated solution to adjust pH to 3.5, cooling at 2 deg.C, standing for 1.5 hr to produce precipitate, centrifuging at 18000 rpm to obtain enzyme paste, and freeze drying to obtain dried bromelain product.
The activity of the bromelain prepared by the process method of the embodiment 1-6 is 140-180 ten thousand, which is far higher than 80 ten thousand of activity of the national standard.
Experimental example 1
This example explores the effect of coagulant release on the precipitation separation of pineapple peel juice.
Experimental group 1 the pour point depressant was added in the manner of inventive example 1, namely polyacrylamide with a molecular weight of 1800 ten thousand was added first, and the addition amount was based on the weight of pineapple peel juice
Stirring the mixtureThen adding high viscosity chitosan solution with molecular weight of 2000 with addition amount of 1.2 ‰, mixing, standing for precipitation for 3 hr, and separating supernatant.
Experiment group 2 only adds polyacrylamide with the molecular weight of 1800 ten thousand as a pour point releasing agent, and the addition amount is the quality of pineapple peel juice
Standing and precipitating after uniformly stirring, and completely precipitating after 6 hours.
Experiment group 3 only adds high viscosity chitosan liquid with molecular weight of 2000 as a releasing agent, the adding amount is 1.2 per mill of the weight of the pineapple peel juice, the mixture is evenly mixed and then stands for precipitation, and the precipitation can be completed after 7 hours.
In the experimental group 4, firstly, a high-viscosity chitosan liquid with molecular weight of 2000 is added as a pour point depressant, the addition amount is 1.2 per thousand of the weight of the pineapple peel juice, after the mixture is uniformly mixed, polyacrylamide with molecular weight of 1800 ten thousand is added as the pour point depressant, and the addition amount is the weight of the pineapple peel juice
Finally, 5 hours are needed for complete precipitation.
The pineapple peel juice in experimental group 4 was not frozen before precipitation, and the addition of the coagulant release agent was the same as in example 1, and precipitation was completed only after 8 h.
In the experimental group 5, no coagulant release agent is added, and the precipitation can be completed after 12 hours.
As can be seen from the experimental examples, the precipitation time is greatly shortened when the combined set-release agent of the invention is added compared with the case that the set-release agent is not added; in addition, the coagulant release agents added in different sequences have a great influence on the sedimentation of the pineapple peel juice.
Experimental example 2
Dividing the concentrated solution after the second-stage ultrafiltration into 5 experimental groups, and respectively precipitating by adopting different precipitation modes, wherein the experimental group 1 adopts an ammonium sulfate salting-out method, specifically adopts 13% of ammonium sulfate by mass and the temperature is 12 ℃; the experimental group 2 adopts a tannin precipitation method, specifically adopts tannin with the mass of 1 per mill, and the temperature is 12 ℃; experiment group 3 used an acid-adjusting isoelectric precipitation method as in example 1; experiment group 4 adopts a CMC precipitation method, specifically adopts 1 per mill of CMC, the temperature is 12 ℃, and the overnight precipitation is carried out; the experimental group 5 employed an alcohol precipitation method, specifically, alcohol with an alcohol content of 50% at a temperature of 12 ℃. The precipitation results are shown in Table 1.
TABLE 1
As can be seen from Table 1, the technical contents of the experimental group 3 (acid-adjusting isoelectric point precipitation method) and the experimental group 4(CMC precipitation method) are higher than those of the experimental group 2 (traditional tannin precipitation method) and the experimental group 1 (ammonium sulfate salting-out method), the enzyme activity is higher, the iron salt content is low, the quality is high, pharmaceutical-grade bromelain can be supported, the clear juice of the bromelain after fruit enzyme extraction can be further comprehensively utilized, the clear juice can be concentrated into a pineapple concentrated juice product, the economic benefit is greatly improved, and the waste liquid pollution is reduced to the maximum extent.
The method comprises the steps of crushing, squeezing, freezing, precipitating, primary ultrafiltration, secondary ultrafiltration, high-speed centrifugation and the like, so that the bromelain is obtained, the steps are simple, no preservative is added in the whole production process, so that the bromelain activity is not lost, the extracted bromelain has the characteristics of high enzyme activity, high purity and less impurities and is far higher than the industrial standard, and the pineapple juice permeate obtained after secondary ultrafiltration can be prepared into a high-concentration pineapple product after twice concentration, so that the pineapple peel can be comprehensively utilized.
The invention freezes the pressed pineapple peel juice, then adds the gelling agent to precipitate the pineapple peel juice at low temperature, can accelerate the aggregation of flocculates in the pineapple peel juice, can accelerate the precipitation speed by adopting a precipitation method of low-temperature standing, and improves the production efficiency of the pineapple enzymes.
The coagulant releasing agent is polyacrylamide and chitosan acetic acid solution with the molecular weight of 1800 ten thousand which are added in sequence, and the flocculation of floccules and impurities in the pineapple enzyme juice can be accelerated by matching the polyacrylamide and the chitosan acetic acid solution and adding the polyacrylamide and the chitosan acetic acid solution in a fixed sequence, so that the precipitation efficiency is improved.
The embodiments described above are some, but not all embodiments of the invention. The detailed description of the embodiments of the present invention is not intended to limit the scope of the invention as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Claims (10)
1. A process for extracting bromelain, which is characterized by comprising the following steps:
cleaning pineapple peel, crushing and squeezing to obtain pineapple peel juice;
freezing pineapple peel juice, adding a coagulant, and precipitating to obtain clear juice;
performing ultrafiltration treatment on the clear juice to obtain a bromelain concentrated solution;
adding acid into the bromelain concentrated solution to adjust pH to acidity, cooling and standing to generate precipitate, centrifuging at high speed to obtain enzyme paste, and freeze drying the enzyme paste to obtain dried bromelain finished product.
2. The process for extracting bromelain according to claim 1, wherein the pineapple peel is crushed to 0.1-0.5 cm.
3. The process for extracting bromelain according to claim 1, wherein the squeezed pineapple peel juice is filtered through a 40-70 mesh sieve.
4. The process for extracting bromelain according to claim 1, wherein the freezing temperature is 2-8 ℃ and the freezing time is 3-12 h.
5. The process for extracting bromelain according to claim 1, wherein the gelling agent comprises a component A and a component B, wherein the component A is polyacrylamide with the molecular weight of 1800 ten thousand, and the ratio of the addition amount of the polyacrylamide to the weight of the pineapple peel juice is (1-1.4): 10000; the component B is chitosan liquid, and the mass ratio of chitosan to pineapple peel juice is (1-1.4): 1000.
6. the process for extracting bromelain according to claim 5, wherein the chitosan solution is prepared by dissolving chitosan with molecular weight of 2000 and high viscosity in dilute acetic acid.
7. The process for extracting bromelain according to claim 1, wherein the ultrafiltration treatment comprises a primary ultrafiltration treatment and a secondary ultrafiltration treatment, the pore size of the ultrafiltration membrane of the primary ultrafiltration treatment is 180000-200000 molecular weight, and the pore size of the ultrafiltration membrane of the secondary ultrafiltration treatment is 5000-6000 molecular weight.
8. The process for extracting bromelain according to claim 1, wherein the acid is hydrochloric acid or acetic acid, and the adjusted pH is 3-5.
9. The process for extracting bromelain according to claim 1, wherein the cooling temperature is 2-4 ℃ and the standing time is 1-2 h.
10. The process for extracting bromelain according to claim 1, wherein the high-speed centrifugation is carried out at 13000-18000 rpm.
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