CN110438110A - Bromelain, its production method and its processing system - Google Patents
Bromelain, its production method and its processing system Download PDFInfo
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- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
- C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
- C12N9/6402—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from non-mammals
- C12N9/6405—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from non-mammals not being snakes
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- C12Y304/22004—Bromelain (3.4.22.4)
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Abstract
The present invention relates to protease to extract field, in particular to a kind of bromelain, its production method and its processing system.A kind of production method of bromelain the following steps are included: to pineapple extract crude liquid be concentrated at least once-mass content of filter operation solid into concentrate is 5-8%；Then, the mass content for carrying out concentration solid into concentrate to concentrate is 18-20%.Crude liquid can be extracted to pineapple by using concentration-filtering be concentrated and be cleaned, while control the mass content of the solid after concentration-filtering in concentrate, can guarantee the purity of bromelain, and guarantee its enzyme activity.Solid content reaches a certain range after concentration-filtering, is no longer filtered, and is only concentrated, and reduces the loss of bromelain, and guarantee enzyme activity.The solid content for controlling final concentrate can guarantee enzyme activity.
Description
Technical field
The present invention relates to protease to extract field, in particular to a kind of bromelain, its production method and its place
Reason system.
Background technique
Bromelain is by the thiol protease of the extractions such as pineapple juice, skin, the extraction side of existing general bromelain
Method is to add benzoic acid, kaolin etc. after fresh pineapple is squeezed the juice to adsorb, and then adjusts pH to 6.7-7.0 and chlorine is then added
Change sodium, then filters pressing, filtrate adjusts pH to 4.8-5.1 again, stands after addition ammonium sulfate and form sediment i.e. bromelain, but on
It is excessively complicated to state extracting method operation, and bromelain extraction efficiency is low, purity is not high, and enzyme activity also reduces relatively.Existing skill
To the improvement of bromelain, there are many methods in art.But some are only capable of improving yield and enzymatic activity is not high, or be only capable of mentioning
Rising enzyme activity, still purity or recovery rate are not promoted, and are handled for example, by using in CN109022402A using ultrafiltration, may
Certain yield can be promoted, but is handled using boiling water, it is more to will lead to the loss of its enzyme activity.
Summary of the invention
The present invention provides a kind of production methods of bromelain, and the technological operation is simple, and extraction process is to enzyme activity shadow
Sound is low, and guarantees extraction efficiency and purity.
The present invention also provides a kind of bromelains, and the bromelain enzyme activity is high, and purity is high has excellent biochemical
Energy.
The present invention also provides a kind of processing system of bromelain, the system can large-scale application in industrial pineapple egg
The extraction of white enzyme, the processing system structure are simple.
The present invention is implemented as follows:
The embodiment of the present invention provides a kind of production method of bromelain, comprising the following steps: extracts crude liquid to pineapple
Be concentrated at least once-mass content of filter operation solid into concentrate is 5-8%；
Then, the mass content for carrying out concentration solid into concentrate to concentrate is 18-20%.
The embodiment of the present invention provides a kind of bromelain, is prepared by the production method of above-mentioned bromelain
It arrives.
The embodiment of the present invention provides a kind of processing system of bromelain, is used to implement the life of above-mentioned bromelain
Production method comprising be concentrated-concentration-filter device of filter operation.
It is concentrated the beneficial effects of the present invention are: the present invention can extract crude liquid to pineapple by using concentration-filtering
And removal of impurities, while the mass content of the solid after concentration-filtering in concentrate is controlled, it can guarantee the purity of bromelain,
And guarantee its enzyme activity.Solid content reaches a certain range after concentration-filtering, is no longer filtered, and is only concentrated, and pineapple egg is reduced
The loss of white enzyme, and guarantee enzyme activity.The solid content for controlling final concentrate can guarantee enzyme activity.
Detailed description of the invention
It, below will be to use required in embodiment in order to illustrate more clearly of the technical solution of embodiment of the present invention
Attached drawing be briefly described, it should be understood that the following drawings illustrates only certain embodiments of the present invention, therefore is not to be seen as
It is the restriction to range, it for those of ordinary skill in the art, without creative efforts, can be with root
Other relevant attached drawings are obtained according to these attached drawings.
Fig. 1 is the structural schematic diagram of the processing system for the bromelain that the embodiment of the present invention 1 provides.
Figure acceptance of the bid note is respectively as follows: the processing system of 100- bromelain；110- cleaning sink；The pre- cold rinse bank of 120-；
130- tears up equipment；140- pocket type juice extractor；The first centrifuge of 150-；160- heat exchanger；170- stands tank；180- second from
Scheming；190- concentration-filter device；191- ultrafiltration apparatus；192- centrifugal device.
Specific embodiment
It in order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below will be in the embodiment of the present invention
Technical solution be clearly and completely described.The person that is not specified actual conditions in embodiment, according to normal conditions or manufacturer builds
The condition of view carries out.Reagents or instruments used without specified manufacturer is the conventional production that can be obtained by commercially available purchase
Product.
A kind of bromelain, its production method and its processing system is provided to the embodiment of the present invention below to illustrate.
The embodiment of the present invention provides a kind of production method of bromelain, comprising the following steps:
Firstly, pineapple successively being cleaned, is crushed, is squeezed the juice and separation of solid and liquid forming the pineapple and extracts crude liquid and precipitating
Object；
What is selected is fresh pineapple rhizome, guarantees that its and enzyme activity high with bromelain content is high.
The operation that cleaning is includes that agitation is cleaned 0.2-0.5 hours in the solution that temperature is 20-30 DEG C by the pineapple,
Then 20-60min is impregnated in the solution that temperature is 8-15 DEG C again；Not only can be into cleaning using aforesaid operations, it can be with
Adequately cooled down to raw material, prevent enzyme activity from losing, and raw material epidermis is made lignifying, the difficulty that the reduction later period is crushed do not occur
Degree, reduces the abrasion of equipment.Solution is generally tap water, other cleaning solutions can also be used.
Agitation cleaning is in 20-30 DEG C of solution raw material to be overturn, and removes the silt on raw material surface, prevents the later period from mentioning
The protease obtained is contaminated.
Further, feeding is screw feeding in cleaning process, and feeding amount is 3-10 ton/hours；Using aforesaid way
Feeding can guarantee the continuity of production, and can guarantee the cleaning effect of every batch materials, promote the subsequent bromelain being prepared
Purity, extraction efficiency and the enzyme activity of enzyme.
Broken includes carrying out Mechanical Crushing, so that enzyme activity loss is less than 0.8%.
It is highly preferred that Mechanical Crushing is cutting attrition crushing.Relatively it is existing beat broken, blade high speed and smash etc. be easy
The more fierce shock method for causing temperature of charge to rise, can prevent temperature of charge liter by specifically cutting attrition crushing
Height, and guarantee broken material suitable size, substantially it is the irregular sheet that size is 1-8 centimetres, reduces enzyme activity loss,
Guarantee that the subsequent bromelain being prepared has high enzyme activity, and guarantees extraction efficiency.
Cutting attrition crushing be using a pair of opposing the knife edge be zigzag circular shear blade, interaction raw material into
Row is squeezed and is cut, and is realized broken.The setting of the circular shear blade can be not only a pair, be also possible to multipair, specific knot
Structure may refer to patent " broken juice device and broken juice system ", the patent No.: ZL201910293774.0.
Juicing includes carrying out mechanical juicing so that crushing juice rate is greater than 60%, and machinery juicing includes that screw rod juicing or pocket type squeeze
Juice, the pressure of machinery juicing are 0.1-0.3MPa.It can guarantee to promote crushing juice rate using above-mentioned juicing mode, then be promoted subsequent
The extraction efficiency of bromelain.
Preferably, being separated by solid-liquid separation includes being centrifuged, and the revolving speed of centrifugation is 1500-3100 revs/min.Centrifuge separation is opposite
Existing plate and frame filtering or standing sedimentation filtering, are able to ascend solid-liquid separation effect, promote the extraction effect of bromelain
Fruit, meanwhile, pollution is reduced, guarantees its purity.
Crude liquid then is extracted to the pineapple to pre-process, and is removed the impurity formed in juice-extracting process, is guaranteed to be prepared into
The purity of the bromelain arrived.
Pretreatment includes the fluid temperature for adjusting the pineapple extraction crude liquid and preliminary removal of impurities；
It is highly preferred that the fluid temperature for adjusting the pineapple extraction crude liquid includes the liquid that the cooling pineapple extracts crude liquid
Temperature so that fluid temperature is 4-10 DEG C, after juicing obtained pineapple extract crude liquid temperature may 20-30 DEG C it
Between, then it is cooled down, enzyme activity loss can be further decreased.
Preliminary removal of impurities includes that the pineapple after adjusting temperature is extracted crude liquid to be stood and be centrifugated；
The time of standing is 12-36h, and the revolving speed of centrifugation is 5000-8000 revs/min.
The cellulose generated in juice-extracting process, macromolecular carbohydrate etc. can be removed using above-mentioned preliminary impurity-removing method, guaranteed
The purity of the subsequent bromelain being prepared.
Then to pineapple extract crude liquid be concentrated at least once-filter operation mass content of solid into concentrate is
5-8%, it is preferable that the mass content of operation solid into concentrate is 7-7.5%.Concentration removal pineapple is extracted molten in crude liquid
Agent promotes the content of the substances such as protein, sugar, fiber in concentrate, then has solid matter precipitation.Inventors have found that
Before the mass content of solid does not reach 5-8% in concentrate, sediment in the concentrate that is concentrated to get, enzyme activity is extremely low,
Substantially it is believed that analyze the overwhelming majority is the impurity such as fiber or starch rather than bromelain, therefore, filtering removal is somebody's turn to do
Partial impurities then guarantee the enzyme activity and purity of the subsequent bromelain being prepared.
Further, concentration-filter operation includes that pineapple extraction crude liquid is successively carried out ultrafiltration and mechanical filter；
And the condition of ultrafiltration are as follows: pressure 0.25-0.35MPa, the membrane aperture that ultrafiltration uses is 10-30DKa；Using above-mentioned
Ultra-filtration conditions can play good concentrated effect, and not will lead to the loss of bromelain, then guarantee bromelain
Extraction efficiency.
Mechanical filter is centrifugal filtration；
It is highly preferred that the condition of centrifugal filtration are as follows: centrifugal rotational speed is 4000-7500 revs/min.
It is filtered using aforesaid way, can guarantee the removal effect of impurity, guarantee the purity and enzyme of bromelain
It is living.
It should be noted that concentration-filtering is will to extract before solid masses content reaches 5-8% in crude liquid in pineapple,
Crude liquid is extracted to pineapple to be concentrated by ultrafiltration, and then carries out centrifugal filtration again.
Further, then, the mass content for carrying out concentration solid into concentrate to concentrate is 18-20%,
Preferably 19%-19.5%.
After the mass content of the solid in concentrate after ultrafiltration reaches 5-8%, then be precipitated sediment enzyme activity it is higher,
Can regard as the sediment being precipitated is bromelain, if continuing to will lead to a large amount of losses of protease using centrifugation, then led
Causing the extraction efficiency of bromelain reduces, therefore, subsequent only to carry out concentration, promotes the extraction effect of bromelain,
And guarantee enzyme activity.The condition being concentrated by ultrafiltration in this step is identical as the condition of above-mentioned ultrafiltration.
And the mass content of solid can guarantee the enzyme activity of protease in the final concentrate of control, advantageously ensure that its yield
And purity.
Further, the mass content of solid is dried final concentrate after reaching 18-20% in final concentrate.
Preferably, dry for freeze-drying；
It is highly preferred that the condition of freeze-drying is quick freezing temperature -40~-35 DEG C, duration 60-90min, vacuum degree
30-40Pa, circulating-heating tube temperature degree are controlled at 15-25 DEG C.
It is dried using freeze-drying, can reduce the enzyme activity loss of bromelain, guarantee enzyme activity, meanwhile, to the greatest extent may be used
It can ground reduction introducing other impurities.
Further, in the extraction process of existing bromelain, squeeze the juice sediment obtained after separation generally as
Waste is lost, studies have found that, much starch, especially amylose are contained in waste, amylose accounts in starch
33.34%.And the amylose in cornstarch only 27%, amylose only accounts for 25% in wheaten starch, in potato starch
Amylose account for the 21.20% of its content, accounting is below pineapple stem starch.Amylose dilated food, health food,
Food additives etc. are widely applied.Therefore, the present invention handles the extraction waste of bromelain, can reduce pineapple
The difficulty of wastewater treatment in protease extraction process, and can economize on resources, and generate additional economic interests.
Further, sediment is post-processed；
Preferably, post-processing includes dry after repeating washing and centrifugally operated to sediment；
It is highly preferred that duplicate number is 1-3 times, drying temperature is 55-70 DEG C.It can be to sediment using aforesaid operations
It is fully handled, guarantees the purity and yield of the starch being prepared.
The embodiment of the present invention also provides a kind of bromelain, is prepared by the production method of above-mentioned bromelain
It arrives；
Preferably, the enzyme activity of the bromelain >=1,630,000 U/g, more preferably ten thousand U/g of 163-200；
Preferably, purity >=90% of the bromelain.
It should be understood that U/g refers to enzyme activity possessed by every gram of zymoprotein.
Below in conjunction with specific embodiment to a kind of bromelain provided by the invention, its production method and its processing system
It is specifically described.
Embodiment 1
The present embodiment provides a kind of production methods of bromelain, comprising the following steps:
1000kg fresh pineapple rhizome (detection feed material enzyme activity 7660U/g) is taken, is put into the water that water temperature is 25 DEG C, turns over
Rolling cleaning 0.5 hour is then sent into the cold water that water temperature is 10 DEG C and impregnates 30min, so that temperature of charge is down to 13 DEG C, feeding amount is
5 ton/hours.Then pineapple rhizome is shredded into fritter, in the irregular sheet of 1-8cm, then plus 4 DEG C of process water 1000kg, stir
30min (sample detection unit enzyme activity 3804U/g), then, then squeezes the juice, and squeeze the juice pressure 0.10MPa, juice about 1250kg
(detection squeezeding juice unit enzyme activity 5964U/g).Squeezeding juice is subjected to centrifugal treating, centrifugal rotational speed is 3100 turns/min, separates shape
Crude liquid 943kg is extracted at sediment 306kg and pineapple.
5 times of water are added in sediment, and wash 2 times, and carry out centrifugal treating after washing every time, finally dries, dries
Temperature be 55 DEG C, obtain pineapple starch 151kg, yield 61%.
Pineapple extracts crude liquid and is cooled down, and after being cooled to 4 DEG C, stands 12h at a temperature of 4 DEG C, and then centrifugal treating removes
76kg is precipitated, remaining 867kg clear liquid carries out film concentration, and centrifugal rotational speed is 6000 turns/min.The operating pressure of film concentration
0.32MPa, ultrafiltration membrane aperture are 20KDA, and concentrate returns after the centrifugation of 5000r/min and carries out film concentration-centrifugation again, are detected
Contain admittedly in concentrate and stops centrifugal treating when reaching 5%.Concentrate only carries out film concentration, contains admittedly until being concentrated into concentrate
Stopper film is concentrated when 19%, obtains concentrate 24.2kg, enzyme activity 310176U/g.Concentrate is freeze-dried, freeze-drying
Condition is -40 DEG C of quick freezing temperature, duration 60min, vacuum degree 30Pa, and circulating-heating tube temperature degree is controlled at 15 DEG C, is finally obtained
Obtain 4.3kg bromelain finished product, 1,630,000 U/g of enzyme activity, enzyme activity yield 91.5%, purity 90%.
The present embodiment provides a kind of bromelains, are prepared by the production method of above-mentioned bromelain,
Enzyme activity is 1,630,000 U/g.
Referring to Fig. 1, the present embodiment also provides a kind of processing system 100 of bromelain, is used to implement above-mentioned pineapple
The production method of protease comprising the cleaning sink 110 and pre- cold rinse bank 120 that pineapple rhizome is cleaned and cooled down, it is right
What pineapple rhizome was torn up tears up equipment 130, the pocket type juice extractor 140 squeezed the juice, first be centrifuged to squeezeding juice
Centrifuge 150, extracts crude liquid to pineapple and carries out cooling heat exchanger 160, extracts the standing tank that crude liquid is stood to pineapple
170, the second centrifuge 180 and carrying out that removal pineapple extracts crude liquid juicing is concentrated-concentration-filter device of filter operation
190；Cleaning sink 110, pre- cold rinse bank 120 tear up equipment 130, pocket type juice extractor 140, the first centrifuge 150, heat exchanger
160, tank 170, the second centrifuge 180 and concentration-filter device 190 is stood to be sequentially connected.
Concentration-filter device 190 includes ultrafiltration apparatus 191 and centrifugal device 192, the ultrafiltration apparatus 191 with it is described from
Center device 192 connects.Ultrafiltration apparatus 191 is connect with the second centrifuge 180.
Embodiment 2
The present embodiment provides a kind of production methods of bromelain, comprising the following steps:
1000kg fresh pineapple rhizome (detection feed material enzyme activity 7500U/g) is taken, is put into the water that water temperature is 30 DEG C, turns over
Rolling cleaning 0.2 hour is then sent into the cold water that water temperature is 15 DEG C and impregnates 60min, so that temperature of charge is down to 17 DEG C, feeding amount is
3 ton/hours.Then pineapple rhizome is shredded into fritter, in the irregular sheet of 1-8cm, then plus 4-10 DEG C of process water 1000kg, turn over
60min (sample detection unit enzyme activity 3752U/g) is stirred, then, then is squeezed the juice, squeeze the juice pressure 0.3MPa, juice about 1397kg
(detection squeezeding juice unit enzyme activity 3633U/g).Squeezeding juice is subjected to centrifugal treating, centrifugal rotational speed is 1500 turns/min, separates shape
Crude liquid 967kg is extracted at sediment 273kg and pineapple.
10 times of water are added in sediment, and wash 2 times, and carry out centrifugal treating after washing every time, finally dries, dries
Temperature be 70 DEG C, obtain pineapple starch 147kg, yield 60%.
Pineapple extracts crude liquid and is cooled down, and after being cooled to 10 DEG C, stands 20h at a temperature of 10 DEG C, then centrifugal treating is gone
Except precipitating 73kg, remaining 889kg clear liquid carries out film concentration, and centrifugal rotational speed is 8000 turns/min.The operating pressure of film concentration
0.35MPa, ultrafiltration membrane aperture are 20KDA, and concentrate returns after the centrifugation of 4000r/min and carries out film concentration-centrifugation again, are detected
Contain admittedly in concentrate and stops centrifugal treating when reaching 8%.Concentrate only carries out film concentration, contains admittedly until being concentrated into concentrate
Stopper film is concentrated when 18%, obtains concentrate 24.9kg, enzyme activity 319990U/g.Concentrate is freeze-dried, freeze-drying
Condition is -35 DEG C of quick freezing temperature, duration 90min, vacuum degree 40Pa, and the control of circulating-heating tube temperature degree is at 25 DEG C, purity
93%.It is final to obtain 4.0kg bromelain finished product, 1,660,000 U/g of enzyme activity, enzyme activity yield 90%, purity 91%.
The present embodiment provides a kind of bromelains, are prepared by the production method of above-mentioned bromelain,
Enzyme activity is 1,660,000 U/g.
Embodiment 3
The present embodiment provides a kind of production methods of bromelain, comprising the following steps:
1000kg fresh pineapple rhizome (detection feed material enzyme activity 7667U/g) is taken, is put into the water that water temperature is 20 DEG C, turns over
Rolling cleaning 0.4 hour is then sent into the cold water that water temperature is 8 DEG C and impregnates 20min, so that temperature of charge is down to 12 DEG C, feeding amount is
10 ton/hours.Then pineapple rhizome is shredded into fritter, in the irregular sheet of 1-8cm, then plus 4-10 DEG C of process water 1000kg,
Stir 60min (sample detection unit enzyme activity 3921U/g), then, then squeezes the juice, and squeeze the juice pressure 0.3MPa, and juice is about
1316kg (detection squeezeding juice unit enzyme activity 6023U/g).Squeezeding juice is subjected to centrifugal treating, 2000 turns/min of centrifugal rotational speed divides
Crude liquid 962kg is extracted from formation sediment 300kg and pineapple.
3 times of water are added in sediment, and wash 3 times, and carry out centrifugal treating after washing every time, finally dries, dries
Temperature be 60 DEG C, obtain pineapple starch 154kg, yield 62%.
Pineapple extracts crude liquid and is cooled down, and after being cooled to 4 DEG C, stands 36h at a temperature of 4 DEG C, and then centrifugal treating removes
85kg is precipitated, remaining 850kg clear liquid carries out film concentration, and centrifugal rotational speed is 5000 turns/min.The operating pressure of film concentration
0.25MPa, ultrafiltration membrane aperture are 30KDA, and concentrate returns after the centrifugation of 7500r/min and carries out film concentration-centrifugation again, are detected
Contain admittedly in concentrate and stops centrifugal treating when reaching 7%.Concentrate only carries out film concentration, contains admittedly until being concentrated into concentrate
Stopper film is concentrated when 20%, obtains concentrate 24.0kg, enzyme activity 3311345U/g.Concentrate is freeze-dried, freeze-drying
Condition be -35 DEG C of quick freezing temperature, duration 60min, vacuum degree 30Pa, circulating-heating tube temperature degree control at 25 DEG C.Finally
Obtain 4.2kg bromelain finished product, 1,810,000 U/g of enzyme activity, enzyme activity yield 93%, purity 95%.
The present embodiment provides a kind of bromelains, are prepared by the production method of above-mentioned bromelain,
Enzyme activity is 1,810,000 U/g.
Embodiment 4
The present embodiment provides a kind of production methods of bromelain, comprising the following steps:
1000kg fresh pineapple rhizome (detection feed material enzyme activity 8000U/g) is taken, is put into the water that water temperature is 25 DEG C, turns over
Rolling cleaning 0,5 hours is then sent into the cold water that water temperature is 12 DEG C and impregnates 60min, so that temperature of charge is down to 14 DEG C, feeding amount is
5 ton/hours.Then pineapple rhizome is shredded into fritter, in the irregular sheet of 1-8cm, then plus 4-10 DEG C of process water 1000kg, turn over
4min (sample detection unit enzyme activity 4076U/g) is stirred, then, then is squeezed the juice, squeeze the juice pressure 0.2MPa, juice about 1291kg
(detection squeezeding juice unit enzyme activity 6121U/g).Squeezeding juice is subjected to centrifugal treating, 3000 turns/min of centrifugal rotational speed, separation is formed
Sediment 312kg and pineapple extract crude liquid 946kg.
5 times of water are added in sediment, and wash 3 times, and carry out centrifugal treating after washing every time, finally dries, dries
Temperature be 65 DEG C, obtain pineapple starch 155kg, yield 63%.
Pineapple extracts crude liquid and is cooled down, and after being cooled to 6 DEG C, stands for 24 hours at a temperature of 6 DEG C, and then centrifugal treating removes
78g is precipitated, remaining 875kg clear liquid carries out film concentration, and centrifugal rotational speed is 5000 turns/min.The operating pressure 0.30MPa of film concentration,
Ultrafiltration membrane aperture is 10KDA, and concentrate returns after the centrifugation of 6000r/min and carries out film concentration-centrifugation again, is detected in concentrate
Gu containing centrifugal treating is stopped when reaching 7.5%.Concentrate only carries out film concentration, until when being concentrated into concentrate admittedly containing 19.5%
Stopper film concentration, obtains concentrate 24kg, enzyme activity 33780U/g.Concentrate is freeze-dried, and the condition of freeze-drying is speed
Freeze -37 DEG C of temperature, duration 70min, vacuum degree 35Pa, circulating-heating tube temperature degree is controlled at 20 DEG C, final to obtain 4.3kg spinach
Trailing plants protease finished product, 1,900,000 U/g of enzyme activity, enzyme activity yield 94%, purity 95%.
The present embodiment provides a kind of bromelains, are prepared by the production method of above-mentioned bromelain,
Enzyme activity is 1,900,000 U/g.
Comparative example 1: the bromelain production method provided according to embodiment 1, processing pineapple rhizome (1000kg, detection
Feed material enzyme activity 7550U/g), squeezeding juice unit enzyme activity 5783U/g, operational difference is that comparative example 1 is mentioned to pineapple
Film concentration is directly carried out after taking crude liquid to be pre-processed, and only carries out film concentration, without concentration-filter operation, until concentrate
Middle solid content is 19%, 1,270,000 U/g of enzyme activity of obtained bromelain, purity 69%, yield 80%.
Comparative example 2: the bromelain production method provided according to embodiment 1, processing pineapple rhizome (1000kg, detection
Feed material enzyme activity 7500U/g), squeezeding juice unit enzyme activity 3752U/g, operational difference is, concentration-mistake in comparative example 2
The mass content of filter operation solid into concentrate is 8%, 1,660,000 U/g of enzyme activity of bromelain, and enzyme activity yield 90% is pure
Degree 91%.
The foregoing is merely the preferred embodiment of the present invention, are not intended to restrict the invention, for this field
For technical staff, the invention may be variously modified and varied.All within the spirits and principles of the present invention, made any
Modification, equivalent replacement, improvement etc., should all be included in the protection scope of the present invention.
Claims (10)
1. a kind of production method of bromelain, which comprises the following steps: extract crude liquid to pineapple and carry out at least
The mass content of primary concentration-filter operation solid into concentrate is 5-8%；
Then, the mass content for carrying out concentration solid into concentrate to concentrate is 18-20%.
2. the production method of bromelain according to claim 1, which is characterized in that the pineapple extract crude liquid into
The mass content of capable concentration-filter operation solid into concentrate at least once is 7-7.5%；
Then, the mass content for carrying out concentration solid into concentrate to concentrate is 19-19.5%.
3. the production method of bromelain according to claim 1, which is characterized in that concentration-filter operation includes will
The pineapple extracts crude liquid and successively carries out ultrafiltration and mechanical filter；
Preferably, the condition of ultrafiltration are as follows: pressure 0.25-0.35MPa, the retaining molecular weight that ultrafiltration uses is 10-30DKa；
Preferably 20KDA,
Preferably, mechanical filter is centrifugal filtration；
It is highly preferred that the condition of centrifugal filtration are as follows: centrifugal rotational speed is 4000-7500 revs/min.
4. the production method of bromelain according to claim 1, which is characterized in that the matter of solid in final concentrate
Amount content is dried final concentrate after reaching 18-20%,
Preferably, dry for freeze-drying；
It is highly preferred that the condition of freeze-drying is quick freezing temperature -35~-40 DEG C, duration 60-90min, vacuum degree 30-
40Pa, circulating-heating tube temperature degree are controlled at 15-25 DEG C.
5. the production method of bromelain according to claim 1, which is characterized in that carrying out that-filter operation is concentrated
It is preceding that pineapple extraction crude liquid is pre-processed,
Preferably, pretreatment includes the fluid temperature for adjusting the pineapple extraction crude liquid and preliminary removal of impurities；
It is highly preferred that the fluid temperature for adjusting the pineapple extraction crude liquid includes the liquid temperature that the cooling pineapple extracts crude liquid
Degree, so that fluid temperature is 4-10 DEG C；
Preliminary removal of impurities includes that the pineapple after adjusting temperature is extracted crude liquid to be stood and be centrifugated；
The time of standing is 12-36h, and the revolving speed of centrifugation is 5000-8000 revs/min.
6. the production method of bromelain according to claim 1, which is characterized in that the pineapple extracts the system of crude liquid
Standby includes: successively to clean pineapple, be crushed, squeezing the juice and separation of solid and liquid forms the pineapple and extracts crude liquid and sediment；
Preferably, the operation of cleaning includes that agitation cleaning 0.2-0.5 is small in the solution that temperature is 20-30 DEG C by the pineapple
When, then 20-60min is impregnated in the solution that temperature is 8-15 DEG C again；Feeding is screw feeding, and feeding amount in cleaning process
It is 3-10 ton/hours；
Preferably, being crushed includes carrying out Mechanical Crushing, so that enzyme activity loss is less than 0.8%；
It is highly preferred that Mechanical Crushing is cutting attrition crushing；
Preferably, juicing includes carrying out mechanical juicing so that crushing juice rate is greater than 60%, and machinery juicing includes screw rod juicing or pocket type
Juicing, the pressure of machinery juicing are 0.1-0.3MPa；
Preferably, being separated by solid-liquid separation includes being centrifuged, and the revolving speed of centrifugation is 1500-3100 revs/min.
7. the production method of bromelain according to claim 1 further includes post-processing to sediment,
Preferably, post-processing includes dry after repeating washing and centrifugally operated to sediment；
It is highly preferred that duplicate number is 1-3 times, drying temperature is 55-70 DEG C.
8. a kind of bromelain, which is characterized in that its production for passing through the described in any item bromelains of claim 1-7
Method is prepared；
Preferably, the enzyme activity of the bromelain >=1,630,000 U/g, more preferably ten thousand U/g of 163-200；
Preferably, purity >=90% of the bromelain.
9. a kind of processing system of bromelain, which is characterized in that it is used to implement the described in any item spinach of claim 1-7
The production method of trailing plants protease comprising be concentrated-concentration-filter device of filter operation.
10. the processing system of bromelain according to claim 9, which is characterized in that the concentration-filter device packet
Ultrafiltration apparatus and centrifugal device are included, the ultrafiltration apparatus is connect with the centrifugal device.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101319208A (en) * | 2008-04-08 | 2008-12-10 | 徐闻县美仑生物制品有限公司 | Method for preparation of bromelin |
| CN101709294A (en) * | 2009-12-10 | 2010-05-19 | 吴茂玉 | Method for purifying bromelin by nano-titanium dioxide porous ceramic column chromatography |
| CN103555695A (en) * | 2013-11-22 | 2014-02-05 | 青岛农业大学 | Method for extracting protease from plant juice |
| CN104611316A (en) * | 2015-02-13 | 2015-05-13 | 广西壮族自治区分析测试研究中心 | Method for extracting bromelin |
| CN108179141A (en) * | 2018-03-28 | 2018-06-19 | 西安瑞仁生物技术有限公司 | A kind of plant rennet extracting method and plant rennet obtained |
| CN109875055A (en) * | 2019-04-12 | 2019-06-14 | 重庆骄王天然产物股份有限公司 | Broken juice device and broken juice system |
-
2019
- 2019-08-14 CN CN201910749988.4A patent/CN110438110B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101319208A (en) * | 2008-04-08 | 2008-12-10 | 徐闻县美仑生物制品有限公司 | Method for preparation of bromelin |
| CN101709294A (en) * | 2009-12-10 | 2010-05-19 | 吴茂玉 | Method for purifying bromelin by nano-titanium dioxide porous ceramic column chromatography |
| CN103555695A (en) * | 2013-11-22 | 2014-02-05 | 青岛农业大学 | Method for extracting protease from plant juice |
| CN104611316A (en) * | 2015-02-13 | 2015-05-13 | 广西壮族自治区分析测试研究中心 | Method for extracting bromelin |
| CN108179141A (en) * | 2018-03-28 | 2018-06-19 | 西安瑞仁生物技术有限公司 | A kind of plant rennet extracting method and plant rennet obtained |
| CN109875055A (en) * | 2019-04-12 | 2019-06-14 | 重庆骄王天然产物股份有限公司 | Broken juice device and broken juice system |
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