CN110438110B - Bromelain, production method thereof and treatment system thereof - Google Patents
Bromelain, production method thereof and treatment system thereof Download PDFInfo
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Abstract
The invention relates to the field of protease extraction, and particularly relates to bromelain, a production method and a treatment system thereof. A method for producing bromelain comprises the following steps: carrying out at least one concentration-filtration operation on the crude pineapple extract until the mass content of solids in the concentrated solution is 5-8%; and then, concentrating the concentrated solution until the mass content of solids in the concentrated solution is 18-20%. The crude pineapple extract can be concentrated and purified by concentration-filtration, and the mass content of solids in the concentrated solution after concentration-filtration is controlled, so that the purity of the bromelain can be ensured, and the enzyme activity of the bromelain can be ensured. After concentration-filtration, the solid content reaches a certain range, filtration is not carried out any more, and only concentration is carried out, so that the loss of bromelain is reduced, and the enzyme activity is ensured. The enzyme activity can be ensured by controlling the solid content of the final concentrated solution.
Description
Technical Field
The invention relates to the field of protease extraction, and particularly relates to bromelain, a production method and a treatment system thereof.
Background
Bromelain is sulfhydryl protease extracted from pineapple juice, peel and the like, and the conventional general extraction method of bromelain is to squeeze fresh pineapples into juice, add benzoic acid, kaolin and the like for adsorption, adjust the pH value to 6.7-7.0, add sodium chloride, filter again, adjust the pH value of filtrate to 4.8-5.1, add ammonium sulfate and stand to form sediment, namely bromelain, but the extraction method is too complex to operate, and has low extraction efficiency, low purity and relatively low enzyme activity. There are many ways to improve bromelain in the prior art. However, some methods only can improve the yield but not the enzyme activity, or only can improve the enzyme activity but not the purity or the extraction rate, for example, the method of treating with the ultrafiltration in CN109022402A may improve a certain yield, but the treatment with boiling water may cause a great loss of the enzyme activity.
Disclosure of Invention
The invention provides a production method of bromelain, the process is simple to operate, the influence of the extraction process on the enzyme activity is low, and the extraction efficiency and the purity are ensured.
The invention also provides bromelain, which has high enzyme activity, high purity and excellent biochemical performance.
The invention also provides a treatment system of the bromelain, which can be applied to the extraction of the bromelain in a large scale and has a simple structure.
The invention is realized by the following steps:
the embodiment of the invention provides a production method of bromelain, which comprises the following steps: carrying out at least one concentration-filtration operation on the crude pineapple extract until the mass content of solids in the concentrated solution is 5-8%;
and then, concentrating the concentrated solution until the mass content of solids in the concentrated solution is 18-20%.
The embodiment of the invention provides bromelain, which is prepared by the production method of bromelain.
The embodiment of the invention provides a treatment system of bromelain, which is used for implementing the production method of bromelain and comprises a concentration-filtration device for performing concentration-filtration operation.
The invention has the beneficial effects that: according to the method, the crude pineapple extract can be concentrated and purified by concentration-filtration, and the mass content of solids in the concentrated solution after concentration-filtration is controlled, so that the purity of bromelain can be ensured, and the enzyme activity of bromelain can be ensured. After concentration-filtration, the solid content reaches a certain range, filtration is not carried out any more, and only concentration is carried out, so that the loss of bromelain is reduced, and the enzyme activity is ensured. The enzyme activity can be ensured by controlling the solid content of the final concentrated solution.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are required 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 schematic structural diagram of a bromelain treatment system provided in example 1 of the present invention.
The labels in the figure are respectively: 100-bromelain treatment system; 110-a washing water tank; 120-precooling a water tank; 130-shredding means; 140-bag juice extractor; 150-a first centrifuge; 160-heat exchanger; 170-standing tank; 180-a second centrifuge; 190-concentration-filtration unit; 191-an ultrafiltration device; 192-centrifugal device.
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.
The following embodiments of the present invention provide a bromelain, a method for producing the same, and a processing system thereof.
The embodiment of the invention provides a production method of bromelain, which comprises the following steps:
firstly, sequentially cleaning, crushing, juicing and carrying out solid-liquid separation on pineapples to form pineapple extract crude liquid and precipitates;
fresh pineapple roots and stems are selected, so that the pineapple has high bromelain content and high enzyme activity.
The cleaning comprises the steps of stirring and cleaning the pineapple in a solution at the temperature of 20-30 ℃ for 0.2-0.5 hour, and then soaking in a solution at the temperature of 8-15 ℃ for 20-60 min; by adopting the operation, the raw materials can be fully cooled, the enzyme activity loss is prevented, the skin of the raw materials is not lignified, the difficulty of later-stage crushing is reduced, and the abrasion of equipment is reduced. The solution is typically tap water, although other cleaning solutions may be used.
The stirring cleaning is to turn over the raw material in a solution at 20-30 ℃ to remove the silt on the surface of the raw material and prevent the protease obtained by later extraction from being polluted.
Further, the feeding in the cleaning process is spiral feeding, and the feeding amount is 3-10 tons/hour; by adopting the feeding mode, the production continuity can be ensured, the cleaning effect of each batch of materials can be ensured, and the purity, the extraction yield and the enzyme activity of the bromelain obtained by subsequent preparation are improved.
The disruption comprises mechanical disruption such that the loss of enzyme activity is less than 0.8%.
More preferably, the mechanical disruption is cutting extrusion disruption. Compared with the existing violent impact methods which easily cause the temperature rise of materials such as hammering crushing, blade high-speed crushing and the like, the temperature rise of the materials can be prevented through specific cutting, extruding and crushing, the crushed materials are guaranteed to be proper in size and basically in irregular sheets with the size of 1-8cm, the enzyme activity loss is reduced, the high enzyme activity of the subsequently prepared bromelain is guaranteed, and the extraction efficiency is guaranteed.
The cutting, extruding and crushing are realized by utilizing a pair of circular blades with saw-tooth-shaped blade points which are oppositely arranged and interacting raw materials to extrude and cut. The arrangement of the circular blades can be not only one pair, but also multiple pairs, and the specific structure can be seen in the patent of crushing and juicing device and crushing and juicing system, and the patent number is as follows: ZL 201910293774.0.
The juicing comprises mechanical juicing to make the juice yield more than 60%, the mechanical juicing comprises screw juicing or bag juicing, and the pressure of the mechanical juicing is 0.1-0.3 MPa. By adopting the juicing mode, the juicing rate can be ensured to be increased, and the extraction yield of the subsequent bromelain is increased.
Preferably, the solid-liquid separation comprises centrifugation at 1500-. Compared with the existing plate-frame filtration or standing sedimentation filtration, the centrifugal separation can improve the solid-liquid separation effect, improve the extraction effect of bromelain, reduce pollution and ensure the purity of bromelain.
And then pretreating the pineapple extracted crude liquid, removing impurities formed in the juicing process, and ensuring the purity of the prepared bromelain.
The pretreatment comprises the steps of adjusting the liquid temperature of the crude pineapple extraction liquid and primarily removing impurities;
more preferably, the step of adjusting the liquid temperature of the crude pineapple extract liquid comprises cooling the liquid temperature of the crude pineapple extract liquid to make the liquid temperature be 4-10 ℃, and the temperature of the crude pineapple extract liquid obtained after juicing is probably 20-30 ℃, and then cooling the crude pineapple extract liquid, so that the enzyme activity loss can be further reduced.
The preliminary impurity removal comprises the steps of standing and centrifugally separating the crude pineapple extraction liquid after the temperature is adjusted;
the standing time is 12-36h, and the rotation speed of the centrifugation is 5000-.
By adopting the preliminary impurity removal method, cellulose, macromolecular saccharides and the like generated in the juicing process can be removed, and the purity of the bromelain prepared subsequently is ensured.
And then carrying out at least one concentration-filtration operation on the crude pineapple extract until the mass content of solids in the concentrated solution is 5-8%, preferably, until the mass content of solids in the concentrated solution is 7-7.5%. Concentrating to remove solvent from the crude pineapple extractive solution, increasing the content of protein, sugar, fiber, etc., and separating out solid substances. The inventor finds that the enzyme activity of the precipitate in the concentrated solution obtained by concentration is extremely low before the mass content of the solid in the concentrated solution reaches 5-8%, and the most of the impurities such as fiber or starch and the like are basically analyzed instead of bromelain, so that the impurities are removed by filtration, and the enzyme activity and the purity of the bromelain obtained by subsequent preparation are ensured.
Further, the concentration-filtration operation comprises sequentially subjecting the crude pineapple extract solution to ultrafiltration and mechanical filtration;
and the ultrafiltration conditions are as follows: the pressure is 0.25-0.35MPa, and the pore diameter of the membrane adopted by ultrafiltration is 10-30DKA; by adopting the ultrafiltration conditions, a good concentration effect can be achieved, the loss of the bromelain can not be caused, and the extraction efficiency of the bromelain is further ensured.
The mechanical filtration is centrifugal filtration;
more preferably, the conditions of the centrifugal filtration are: the centrifugal speed was 4000 and 7500 rpm.
By adopting the method for filtering, the impurity removal effect can be ensured, and the purity and the enzyme activity of the bromelain can be ensured.
The concentration-filtration is to perform ultrafiltration concentration on the crude pineapple extract liquid before the mass content of solids in the crude pineapple extract liquid reaches 5-8%, and then perform centrifugal filtration.
Further, concentrating the concentrated solution until the mass content of solids in the concentrated solution is 18-20%, preferably 19-19.5%.
When the mass content of solids in the concentrated solution after ultrafiltration reaches 5-8%, the enzyme activity of the precipitate separated out is high, the separated precipitate can be determined as bromelain, if centrifugation is continuously adopted, a large amount of protease loss can be caused, and then the extraction efficiency of the bromelain is reduced, so that only concentration treatment is subsequently performed, the extraction effect of the bromelain is improved, and the enzyme activity is ensured. The conditions for ultrafiltration concentration in this step are the same as those for the above-mentioned ultrafiltration.
And the enzyme activity of the protease can be ensured by controlling the mass content of the solid in the final concentrated solution, and the yield and the purity of the protease can be ensured.
Further, drying the final concentrated solution after the mass content of the solid in the final concentrated solution reaches 18-20%.
Preferably, the drying is freeze-drying;
more preferably, the conditions of freeze drying are that the quick freezing temperature is-40 to-35 ℃, the duration is 60 to 90min, the vacuum degree is 30 to 40Pa, and the temperature of the circulating heating pipe is controlled to be 15 to 25 ℃.
The freeze drying is adopted for drying, so that the enzyme activity loss of the bromelain can be reduced, the enzyme activity is ensured, and other impurities are reduced as much as possible.
Further, in the existing extraction process of bromelain, the precipitate obtained after juicing and separating is generally discarded as waste, but researches show that the waste contains a large amount of starch, particularly amylose, and the amylose in the starch accounts for 33.34%. The amylose in the corn starch is only 27 percent, the amylose in the wheat starch is only 25 percent, and the amylose in the potato starch is 21.20 percent of the content of the amylose, and the proportion of the amylose is lower than that of the pineapple stem starch. Amylose is widely applied to puffed food, health-care food, food additives and the like. Therefore, the method treats the extraction waste of the bromelain, can reduce the difficulty of wastewater treatment in the bromelain extraction process, can save resources and generate additional economic benefits.
Further, post-treating the precipitate;
preferably, the post-treatment comprises drying the precipitate after repeating the water washing and centrifuging operations;
more preferably, the number of repetitions is 1 to 3 and the drying temperature is 55 to 70 ℃. By adopting the operation, the precipitate can be fully treated, and the purity and the yield of the prepared starch are ensured.
The embodiment of the invention also provides bromelain, which is prepared by the production method of the bromelain;
preferably, the enzymatic activity of the bromelain is more than or equal to 163 ten thousand U/g, more preferably 163- & lt 200 & gtU/g;
preferably, the purity of the bromelain is more than or equal to 90%.
It should be noted that: u/g refers to the enzyme activity per gram of enzyme protein.
The following will specifically describe a bromelain, its production method and its processing system according to the present invention with reference to specific examples.
Example 1
This example provides a method for producing bromelain, comprising the following steps:
1000kg of fresh pineapple roots and stems (the unit enzyme activity of the detected raw materials is 7660U/g) are taken and put into water with the water temperature of 25 ℃, the pineapple roots and the stems are rolled and cleaned for 0.5 hour, then the pineapple roots and the stems are sent into cold water with the water temperature of 10 ℃ to be soaked for 30min, the temperature of the materials is reduced to 13 ℃, and the feeding amount is 5 tons/hour. Then the pineapple rootstock is shredded into small pieces which are irregular sheets of 1-8cm, 1000kg of process water with the temperature of 4 ℃ is added, stirring is carried out for 30min (3804U/g of enzyme activity per unit of sampling and detecting), then juicing is carried out, the juicing pressure is 0.10MPa, and the juice is about 1250kg (5964U/g of enzyme activity per unit of detecting juice). Centrifuging the juice at 3100 rpm to separate precipitate 306kg and crude pineapple extractive solution 943 kg.
And adding 5 times of water into the precipitate, washing for 2 times, performing centrifugal treatment after each washing, and finally drying at the temperature of 55 ℃ to obtain 151kg of pineapple starch with the yield of 61%.
Cooling the crude pineapple extract, cooling to 4 ℃, standing at 4 ℃ for 12h, then centrifuging to remove 76kg of precipitate, and performing membrane concentration on the residual 867kg of clear liquid at the centrifugal rotating speed of 6000 r/min. The working pressure of the membrane concentration is 0.32MPa, the aperture of the ultrafiltration membrane is 20KDA, the concentrated solution is centrifuged at 5000r/min and then returned for membrane concentration-centrifugation, and the centrifugation treatment is stopped when the solid content in the concentrated solution is detected to reach 5 percent. The concentrated solution is only subjected to membrane concentration until the concentrated solution is concentrated to 19 percent of solid content, and the membrane concentration is stopped to obtain 24.2kg of concentrated solution with enzyme activity of 310176U/g. And (3) freeze-drying the concentrated solution, wherein the freeze-drying conditions comprise a quick-freezing temperature of-40 ℃, a duration of 60min, a vacuum degree of 30Pa and a temperature of a circulating heating pipe controlled at 15 ℃, and finally obtaining 4.3kg of a finished product of the bromelain, the enzyme activity of 163 ten thousand U/g, the enzyme activity yield of 91.5 percent and the purity of 90 percent.
The embodiment provides bromelain, which is prepared by the production method of bromelain, and the enzyme activity of the bromelain is 163 ten thousand U/g.
Referring to fig. 1, the present embodiment further provides a bromelain processing system 100 for implementing the above-mentioned method for producing bromelain, which includes a cleaning water tank 110 and a pre-cooling water tank 120 for cleaning and cooling pineapple roots, a shredding device 130 for shredding pineapple roots, a bag juicer 140 for squeezing juice, a first centrifuge 150 for centrifuging juice, a heat exchanger 160 for cooling crude pineapple extract, a standing tank 170 for standing crude pineapple extract, a second centrifuge 180 for removing crude pineapple extract and squeezing juice, and a concentration-filtration device 190 for performing concentration-filtration operation; the wash tank 110, the pre-cooling tank 120, the shredding device 130, the bag juicer 140, the first centrifuge 150, the heat exchanger 160, the resting tank 170, the second centrifuge 180, and the concentration-filtration device 190 are connected in sequence.
The concentration-filtration device 190 comprises an ultrafiltration device 191 and a centrifugation device 192, the ultrafiltration device 191 being connected to the centrifugation device 192. The ultrafiltration device 191 is connected to a second centrifuge 180.
Example 2
This example provides a method for producing bromelain, comprising the following steps:
1000kg of fresh pineapple roots and stems (7500U/g of unit enzyme activity of the raw materials) are taken and put into water with the water temperature of 30 ℃, rolled and cleaned for 0.2 hour, and then the fresh pineapple roots and the stems are put into cold water with the water temperature of 15 ℃ for soaking for 60 minutes, so that the temperature of the materials is reduced to 17 ℃, and the feeding amount is 3 tons/hour. Then the pineapple rootstock is shredded into small pieces which are irregular sheets of 1-8cm, 1000kg of process water of 4-10 ℃ is added, stirring is carried out for 60min (unit enzyme activity is detected by sampling and detecting 3752U/g), then juice is squeezed again, the pressure of the juice squeezing is 0.3MPa, and the juice is about 1397kg (unit enzyme activity is detected by detecting juice squeezing unit 3633U/g). Centrifuging the juice at 1500 rpm to obtain 273kg precipitate and 967kg crude pineapple extractive solution.
Adding 10 times of water into the precipitate, washing for 2 times, centrifuging after each washing, and finally drying at 70 ℃ to obtain 147kg of pineapple starch with the yield of 60%.
Cooling the crude pineapple extract, cooling to 10 ℃, standing at 10 ℃ for 20h, centrifuging to remove 73kg of precipitate, and performing membrane concentration on the residual 889kg of clear liquid at the centrifugal rotation speed of 8000 rpm. The working pressure of the membrane concentration is 0.35MPa, the aperture of the ultrafiltration membrane is 20KDA, the concentrated solution is centrifuged at 4000r/min and then returned for membrane concentration-centrifugation, and the centrifugation treatment is stopped when the solid content in the concentrated solution is detected to reach 8 percent. The concentrated solution is only subjected to membrane concentration until the concentrated solution is concentrated to 18 percent of solid content, and the membrane concentration is stopped to obtain 24.9kg of concentrated solution with enzyme activity of 319990U/g. Freeze drying the concentrated solution at-35 deg.C for 90min under vacuum degree of 40Pa, and circulating heating tube at 25 deg.C and 93% purity. Finally obtaining 4.0kg of bromelain finished product, wherein the enzyme activity is 166 ten thousand U/g, the enzyme activity yield is 90 percent, and the purity is 91 percent.
The embodiment provides bromelain, which is prepared by the production method of bromelain, and the enzyme activity of the bromelain is 166 ten thousand U/g.
Example 3
This example provides a method for producing bromelain, comprising the following steps:
1000kg of fresh pineapple roots and stems (the unit enzyme activity of the detected raw materials is 7667U/g) are taken and put into water with the water temperature of 20 ℃, the pineapple roots and the stems are rolled and cleaned for 0.4 hour, then the pineapple roots and the stems are sent into cold water with the water temperature of 8 ℃ to be soaked for 20min, the temperature of the materials is reduced to 12 ℃, and the feeding amount is 10 tons/hour. Then, the pineapple rootstocks are shredded into small pieces which are irregular sheets of 1-8cm, 1000kg of process water with the temperature of 4-10 ℃ is added, stirring is carried out for 60min (the unit enzyme activity of sampling detection is 3921U/g), then, juice squeezing is carried out, the juice squeezing pressure is 0.3MPa, and the juice yield is about 1316kg (the unit enzyme activity of juice squeezing is 6023U/g). Centrifuging the juice at 2000 rpm to obtain precipitate 300kg and crude pineapple extractive solution 962 kg.
Adding 3 times of water into the precipitate, washing for 3 times, centrifuging after each washing, and finally drying at 60 ℃ to obtain 154kg of pineapple starch with the yield of 62%.
Cooling the crude pineapple extract, cooling to 4 ℃, standing at 4 ℃ for 36h, then centrifuging to remove 85kg of precipitate, and performing membrane concentration on the residual 850kg of clear liquid at the centrifugal rotation speed of 5000 r/min. The working pressure of the membrane concentration is 0.25MPa, the aperture of the ultrafiltration membrane is 30KDA, the concentrated solution is centrifuged at 7500r/min and then returned for membrane concentration-centrifugation, and the centrifugation treatment is stopped when the solid content in the concentrated solution is detected to reach 7 percent. The concentrated solution is only subjected to membrane concentration until the concentrated solution is concentrated to 20 percent of solid content, and the membrane concentration is stopped to obtain 24.0kg of concentrated solution with enzyme activity of 3311345U/g. Freeze drying the concentrated solution at-35 deg.C for 60min under vacuum degree of 30Pa, and controlling the temperature of the circulating heating pipe at 25 deg.C. Finally obtaining 4.2kg of bromelain finished product, wherein the enzyme activity is 181 ten thousand U/g, the enzyme activity yield is 93 percent, and the purity is 95 percent.
The embodiment provides bromelain, which is prepared by the production method of bromelain, and the enzyme activity of the bromelain is 181 ten thousand U/g.
Example 4
This example provides a method for producing bromelain, comprising the following steps:
1000kg of fresh pineapple rhizomes (8000U/g of enzyme activity of a unit of detection raw materials) are taken and put into water with the water temperature of 25 ℃, the pineapple rhizomes are rolled and cleaned for 0 to 5 hours, then the pineapple rhizomes are sent into cold water with the water temperature of 12 ℃ to be soaked for 60 minutes, the temperature of the materials is reduced to 14 ℃, and the feeding amount is 5 tons/hour. Then, the pineapple roots and stems are shredded into small pieces which are irregular sheets of 1-8cm, 1000kg of process water of 4-10 ℃ is added, stirring is carried out for 4min (4076U/g of unit enzyme activity in sampling and detecting), then, juice is squeezed again, the pressure of the juice squeezing is 0.2MPa, and the juice yield is about 1291kg (6121U/g of unit enzyme activity in juice squeezing). Centrifuging the juice at 3000 rpm to separate precipitate 312kg and crude pineapple extractive solution 946 kg.
And adding 5 times of water into the precipitate, washing for 3 times, performing centrifugal treatment after each washing, and finally drying at the temperature of 65 ℃ to obtain 155kg of pineapple starch with the yield of 63%.
Cooling the crude pineapple extract, cooling to 6 ℃, standing for 24h at 6 ℃, then centrifuging to remove 78g of precipitate, and performing membrane concentration on the residual 875kg of clear liquid at the centrifugal rotation speed of 5000 r/min. The working pressure of the membrane concentration is 0.30MPa, the aperture of the ultrafiltration membrane is 10KDA, the concentrated solution is centrifuged at 6000r/min and then returned to carry out the membrane concentration-centrifugation, and the centrifugation treatment is stopped when the solid content in the concentrated solution is detected to reach 7.5 percent. The concentrated solution is only subjected to membrane concentration until the concentrated solution is concentrated to 19.5 percent of solid content, and the membrane concentration is stopped to obtain 24kg of concentrated solution with the enzyme activity of 33780U/g. And (3) freeze-drying the concentrated solution, wherein the freeze-drying conditions comprise a quick-freezing temperature of-37 ℃, a duration of 70min, a vacuum degree of 35Pa, and a temperature of a circulating heating pipe controlled at 20 ℃, so that 4.3kg of a finished product of the bromelain is finally obtained, the enzyme activity is 190 ten thousand U/g, the enzyme activity yield is 94%, and the purity is 95%.
The embodiment provides bromelain, which is prepared by the production method of bromelain, and the enzyme activity of the bromelain is 190 ten thousand U/g.
Comparative example 1: according to the method for producing bromelain provided by the example 1, the pineapple roots and stems (1000kg, 7550U/g of enzyme activity of raw material unit is detected) are treated, and 5783U/g of enzyme activity of juice unit is detected, the difference in operation lies in that in the comparative example 1, membrane concentration is directly carried out after the crude pineapple extract is pretreated, and only membrane concentration is carried out, and concentration-filtration operation is not carried out, so that the solid content in the concentrated solution is 19%, the enzyme activity of the obtained bromelain is 127 ten thousand U/g, the purity is 69%, and the yield is 80%.
Comparative example 2: according to the method for producing bromelain provided in example 1, the pineapple roots and stems (1000kg, 7500U/g of enzyme activity per unit of detected raw material) are treated, and 3752U/g of enzyme activity per unit of juice is treated, wherein the difference in operation lies in that in comparative example 2, concentration and filtration are carried out until the mass content of solids in the concentrated solution is 8%, the enzyme activity of bromelain is 166 ten thousand U/g, the enzyme activity yield is 90%, and the purity is 91%. .
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (8)
1. A method for producing bromelain, which is characterized by comprising the following steps: carrying out at least one concentration-filtration operation on the crude pineapple extract until the mass content of solids in the concentrated solution is 5-8%;
then, concentrating the concentrated solution until the mass content of solids in the concentrated solution is 18-20%;
the concentration-filtration operation comprises sequentially subjecting the crude pineapple extract to ultrafiltration and mechanical filtration;
wherein, the ultrafiltration conditions are as follows: the pressure is 0.25-0.35MPa, the molecular weight of the membrane used for ultrafiltration is 10-30DKA, the mechanical filtration is centrifugal filtration, and the centrifugal rotation speed is 4000 and 7500 r/min;
pre-treating the crude pineapple extract liquid before concentration-filtration operation, wherein the pre-treatment comprises adjusting the liquid temperature of the crude pineapple extract liquid and primarily removing impurities;
adjusting the liquid temperature of the crude pineapple extract liquid comprises cooling the liquid temperature of the crude pineapple extract liquid to a liquid temperature of 4-10 ℃;
the preliminary impurity removal comprises the steps of standing and centrifugally separating the crude pineapple extraction liquid after the temperature is adjusted; the standing time is 12-36h, and the centrifugal rotating speed is 8000 rpm and 5000-;
the preparation method of the crude pineapple extract liquid comprises the following steps: sequentially cleaning, crushing, juicing and carrying out solid-liquid separation on pineapples to form pineapple extraction crude liquid and precipitates;
the cleaning operation comprises stirring and cleaning the pineapple in a solution at 20-30 deg.C for 0.2-0.5 hr, and soaking in a solution at 8-15 deg.C for 20-60 min; the feeding in the cleaning process is spiral feeding, and the feeding amount is 3-10 tons/hour;
the crushing comprises mechanical crushing, so that the enzyme activity loss is less than 0.8%; the mechanical crushing is cutting extrusion crushing;
the juicing comprises mechanical juicing to make the juice yield more than 60%, wherein the mechanical juicing comprises screw juicing or bag juicing, and the pressure of the mechanical juicing is 0.1-0.3 MPa;
the solid-liquid separation comprises centrifugation, and the rotation speed of the centrifugation is 1500-3100 rpm.
2. The method for producing bromelain according to claim 1, wherein the crude pineapple extract is subjected to at least one concentration-filtration operation until the mass content of solids in the concentrated solution is 7-7.5%;
and then, concentrating the concentrated solution until the mass content of solids in the concentrated solution is 19-19.5%.
3. The method for producing bromelain according to claim 1, wherein ultrafiltration is carried out using a membrane having a molecular weight cut-off of 20 kDa.
4. The method of claim 1, wherein the final concentrate is dried after the mass content of solids in the final concentrate reaches 18-20%.
5. The method for producing bromelain according to claim 4, wherein the drying is freeze-drying.
6. The method for producing bromelain according to claim 5, wherein the conditions of freeze drying are quick freezing temperature of-35 to-40 ℃, duration of 60 to 90min, vacuum degree of 30 to 40Pa, and temperature of a circulating heating pipe controlled at 15 to 25 ℃.
7. The method for producing bromelain according to claim 1, further comprising post-treating the precipitate.
8. The method for producing bromelain according to claim 7, wherein the post-treatment comprises drying the precipitate after repeating the water washing and the centrifugation; the repetition times are 1-3 times, and the drying temperature is 55-70 ℃.
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