CN109022402A - A kind of extracting method of bromelain - Google Patents
A kind of extracting method of bromelain Download PDFInfo
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- CN109022402A CN109022402A CN201810953068.XA CN201810953068A CN109022402A CN 109022402 A CN109022402 A CN 109022402A CN 201810953068 A CN201810953068 A CN 201810953068A CN 109022402 A CN109022402 A CN 109022402A
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
- A23L27/10—Natural spices, flavouring agents or condiments; Extracts thereof
- A23L27/11—Natural spices, flavouring agents or condiments; Extracts thereof obtained by solvent extraction
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
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- C12Y—ENZYMES
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- 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 present invention provides a kind of extracting method of bromelain, operating procedures are as follows: 1) pre-processes;2) it digests;3) isoelectric precipitation;4) ultrafiltration;5) kept dry.It has the beneficial effect that and pineapple or pineapple stem is first subjected to precooling treatment, can be reduced the loss of bromelain enzyme activity;The stabilizer of use can enhance the stability of enzyme texture image, improve the storage-stable of enzyme;With preferable inoxidizability, free radical is removed, protects sulfydryl;There is complexing to heavy metal, prevent heavy metal from inhibiting the activity of bromelain;The indexs such as enzyme activity, bacterial content, content of beary metal, the appearance of the bromelain finally obtained meet or exceed medical grade standard;Operating procedure is simple, low in cost, is suitble to large-scale production.
Description
Technical field
The present invention relates to protease extractive technique field, the extracting methods of specifically a kind of bromelain.
Background technique
Bromelain (bromelain, EC3.4.22.33) is to be present in pineapple plant (including fruit, stem, handle and blade)
In proteolytic enzyme, have multiple isodynamic enzymes, sulfydryl (- SH) is contained in activated centre, so belong to thiol protease.Pineapple egg
White enzyme is widely used in the industries such as medicine, food, cosmetics, weaving, feed, leather.It can decompose fibrin, hemoglobin etc.
Protein can make fibrin and dissolution of blood clot in the lesser situation of side effect, and then improve body fluid circulatory, increase tissue
Permeability leads to inflammation, edema extinction;The infiltration and diffusion that can promote drug to lesion are shared with antibiotic, chemotherapeutics, from
And it heightens the effect of a treatment.
The method for extraction and purification of protein is mainly saltoutd, isoelectric precipitation, organic solvent deposit, tannin (tannic acid) are heavy
The methods of shallow lake, ion-exchange chromatography, affinity chromatography, ultrafiltration concentration.The production technology of bromelain mainly has ammonium sulfate precipitation
Method, the tannin precipitation method, ultrafiltration etc., these methods have its disadvantage: ammonium sulfate precipitation method is needed using a large amount of ammonium sulfate, ring
Border is seriously polluted;The resulting enzyme preparation of the tannin precipitation method is insoluble in water, and enzyme activity is low;Equipment investment needed for ultrafiltration is big, pineapple
Viscosity is gradually increased during juice is concentrated by ultrafiltration, and ultrafiltration membrane easily blocks, therefore the production cycle is long, and extract yield is low.
The prior art such as Authorization Notice No. is the Chinese invention patent of CN103305492B, discloses a kind of bromelain
Manufacturing method, especially a kind of extracting method of high-purity pharmaceutical grade bromelain.It is the following steps are included: pineapple is located in advance
Reason, coarse crushing, enzymatic hydrolysis, juicing, filtering, addition auxiliary material, essence filtering, addition stabilizer, adsorbent absorption, elution, ultrafiltration, freezing
It is dry to obtain finished product.The advantages of this method is: replacing kaolin in traditional bromelain extraction using cellulose absorption method
Absorption method, the tannin precipitation method, organic solvent extractionprocess, yield height, simple process, low energy consumption, bromelain enzyme produced
The indexs such as vigor, bacterial content, content of beary metal, appearance meet or exceed medical grade standard.In addition, adsorptive cellulose is through eluting
It may be reused after regeneration, production cost can greatly be reduced.But it is adsorbed using cellulose, the recovery rate of bromelain
Low, loss late is high.
As people pursue health, advocate the continuous enhancing of the natural consumption consciousness, integrate safe and healthy, naturally
The development of natural flavouring have received widespread attention and pay attention to.Natural flavouring mainly includes that extract series seasoning (mentions
Be derived from livestock products, aquatic products, greengrocery and yeast), digest serial seasoning and fish sauce flavourings etc..Wherein, natural extract
Class seasoning is mainly using natural animal-plant as raw material, with modern extraction process, in conjunction with microencapsulation, spraying, vacuum drying
Etc. new technologies be process.Natural flavouring is divided into breakdown type natural flavouring and extracting type seasoning again.Breakdown type is naturally adjusted
Taste substance includes sour breakdown type and enzyme breakdown type.Sour breakdown type is with the vegetable proteins such as defatted soybean, corn gluten raw material and fish
The protein hydrolysate (HVP, HAP) that the animal protein such as powder, animal glue raw material is process through sour water solution, containing various amino acid and
Peptides, although the flavor of original meat and vegetables is difficult to retain, are increasing taste with the flavor that flavor is complicated and haves a wide reach
But there is certain effect in terms of the dense degree in road, adjustment taste, less salt can be played, slow down bitter taste, remove and detest taste and other effects.Enzyme point
Solution type is mainly used for the preparation of yeast extract and meat original powder.Yeast extract is that yeast cells internal protein is degraded into amino acid
And polypeptide, nucleolysis are nucleotide, and them and other effective components such as B family vitamin, glutathione, microelement etc.
Soluble nutrition and flavor that the obtained human body of discharging can directly be absorbed and utilized are extracted from yeast cells with equipment and instrument
The concentrate of matter.It has the unexistent feature of many other seasonings, has and alleviates tart flavour, except bitter taste and other effects, different to peculiar smell
It is smelly that there is shielding action.Extracting type seasoning: the type seasoning is the heated extracting of the natural materials such as livestock products, aquatic products, squeezing
Juice, enzyme the processes purification processing such as connect and are concentrated.
Chinese patent 201410209287.9 discloses a kind of natural nutrition seafood condiment for not adding preservative, by weight
Measuring part includes following components: Optimization of Low Value Fish enzymolysis liquid 100;Sweetener 2-20;Fumet 0.01-10;Acid 0.01-0.05.It should
Production method: the A, preparation of Optimization of Low Value Fish enzymolysis liquid;B, the preparation of natural nutrition seafood condiment: into Optimization of Low Value Fish enzymolysis liquid by than
Example is separately added into sweetener, fumet, acid, thickener, colorant, and heating is concentrated into thickener and pastes completely after mixing evenly
After change it is filling while hot to get.The purpose of the invention is place in order to overcome the shortcomings in the prior art, provides a kind of letter of component
List, produces simple and convenient, natural nutrition, natural nutrition seafood condiment that is tasteful, not adding preservative at quality safety.
Chinese patent 201110335768.0 discloses one kind and removes flavor and enhancing flavor compound seasoner, it is in traditional spice base
It is improved on plinth, the removal taste weight excessively spice of stimulation and the classifications such as biggish Chinese prickly ash of irritation, selected addition
The natural Chinese medicinal herbs such as rhizoma nardostachyos, Randia cochinchinensis, Radix Angelicae Sinensis have determined that these Chinese herbal medicines and traditional perfume are pungent after having found after test many times
The optimal components ratio of material combines, and has not only been sufficiently reserved the seasoning function of traditional spice, but also ideally played the characteristic of Chinese herbal medicine,
Provide that a kind of taste is unique, flavor and enhancing flavor compound seasoner of dispelling of natural health for people.The invention dispel raw meat, dispel have a strong smell, flavouring
The effect of, it is vdiverse in function, easy to use, it may replace the common five-spice powder of person cooks, halogen powder, fricassee powder, barbecue material etc..
Chinese patent 201410346150.8 discloses a kind of production technology of new type natural extract seasoning material, including
Following steps: pretreatment, high temperature and pressure boiling sterilization, miniaturization processing, enzymatic hydrolysis, concentration, Maillard reaction.High temperature high pressure process
Most protein and fat are dissolved out, then is converted into the nutrient and flavours such as small molecule physiologically active peptide and amino acid through enzyme digestion reaction
Ingredient, finally by the further flavouring flavour enhancing of Maillard reaction, obtained new type natural extract seasoning item matter height, mouth
Feel, delicate flavour is splendid, full of nutrition, health index is high.
In the prior art, natural seasoning focuses on the mouthfeel of seasoning more, fresh perfume taste or its flavouring, dispel have a strong smell,
The effect of flavouring, however when preparing natural flavouring as raw material with meat, marine product, since the seasoning of preparation contains amino more
The physiological activators such as acid, polypeptide, taurine, active peptide, nucleotide, using prepared by inappropriate processing and stored manner
Seasoning be easy it is putrid and deteriorated, it is therefore desirable to design a kind of seasoning composite extract, guaranteeing good flavouring effect
While can also inhibit the growth of microorganism.
Summary of the invention
The purpose of the present invention is to provide a kind of recovery rate height, organic solvent is not used, environmentally protective, the pineapple egg of extraction
White enzyme purity is high, enzyme activity is strong, the extracting method of bacterium and the seldom bromelain of content of beary metal.
The present invention in background technique aiming at the problem that mentioning, the technical solution taken are as follows:
A kind of extracting method of bromelain, specific steps are as follows:
1) it pre-processes: fresh pineapple or pineapple stem being cleaned, precooling treatment, the temperature of precooling treatment is 0~2 DEG C, pre-cooling
Time is 21~25h.It crushes, juicing.Pineapple or pineapple stem are first subjected to precooling treatment, can be reduced bromelain enzyme activity damage
It loses;
2) it digests: distilled water, lignoenzyme and stabilizer, the pineapple with pulp being added in the pineapple juice with pulp
Juice: distilled water mass ratio is 1:6~10.Lignoenzyme additional amount is 0.2~0.4%, the additional amount of stabilizer is 0.004~
0.007%, pH are 3.5~4.2, and temperature is 38~43 DEG C, and enzymolysis time is 3~3.4h, and boiling water bath enzyme deactivation after enzymatic hydrolysis is adjusted
PH is saved, 0.3~0.5% cellulase and 0.1~0.2% pectase is added, hydrolysis temperature is 53~56 DEG C, pH is 2.7~
3.2, enzymolysis time is 2.0~2.3h.Boiling water bath enzyme deactivation after enzymatic hydrolysis, filtering.Lignoenzyme hydrolyzes lignin, and pineapple is thin
Born of the same parents lose support, spread out, and increase the contact area with enzyme solution;Cellulase destroys the cell wall of pineapple cell, makes pineapple
Cytoplasm outflow;Pectase continues lysed cells matter, makes bromelain dispersion in aqueous solution, increases mentioning for bromelain
Take rate;
3) isoelectric precipitation: adjusting filtrate pH value is 9.5~9.6, is stirred, and is stood, filtering.Pineapple when pH is 9.5~9.6
Protease is in electroneutral, and solubility in water sharply declines, and makees initial gross separation purifying to bromelain;
4) ultrafiltration: being added distilled water in precipitating, ultrafiltration, first with the ultrafiltration membrane ultrafiltration of 6~8kDa, takes filtered solution, then with 0.9
The ultrafiltration membrane ultrafiltration of~1.2kDa, takes stagnant liquid.Successively removal molecular weight is greater than 6~8kDa, molecular weight less than 0.9~1.2kDa
Impurity;It desalts concentration to bromelain enzyme solutions, shortens drying time, save the energy;
5) kept dry: stagnant liquid concentrated by rotary evaporation is freeze-dried in -40~-50 DEG C, and stabilizer is added in grinding, is stablized
Agent: bromelain powder volume ratio is 1:300~400, and stirring obtains bromelain.The bromelain finally obtained
The indexs such as enzyme activity, bacterial content, content of beary metal, appearance meet or exceed medical grade standard.
Preferably, stabilizer composition and its parts by weight are as follows: 110~20 parts of B-1 stabilizers, 0.004~0.005 portion of eggplant are red
Element, 2~5 parts of EDTA, 0.002~0.005 part of inclined sulfurous acid and 0.001~0.003 part of folic acid.Sulfydryl, amino, trp residue
It is bromelain enzymatic activity essential group with histidine residues.The variation of the microenvironment of the conformation and catalytic site of enzyme molecule
Influence the catalysis activity of enzyme.Above stabilizer has embedding effect, isolation from oxygen and water to bromelain, at the same with protein shape
At hydrogen bond.Since oxygen has been isolated, sulfydryl can be prevented to be oxidized, the hydrogen bond of formation enhances the stability of enzyme texture image;Albumen
Matter interaction area domain separation water, reduces free energy, the storage-stable of enzyme is made to have obtained significant raising;With preferable
Inoxidizability, remove free radical, protect sulfydryl;Folic acid and Lycopene are played by inclined sulfurous acid, EDTA to be promoted to heavy metal
Complexing, prevent heavy metal inhibit bromelain activity.
Preferably, with one of sodium hydroxide, sodium carbonate, sodium bicarbonate, potassium hydroxide, potassium carbonate, saleratus or
A variety of adjusting pH value of solution.Heavy metal or other, which will not be introduced, can inhibit the active substance of bromelain, and it is good to adjust pH effect.
The present invention can summarize are as follows:
The object of the present invention is to provide a kind of preparation methods of composite flavored extract, specifically includes the following steps:
Fresh bonito stick is chosen, break process is carried out, it is spare;Tissue mashing machine, mashing, by enzyme is added in bonito stick after break process
Solution, obtains enzymolysis liquid;Ginger powder, garlic powder, fennel seeds, mulberry leaf, Ground Cloves, salt and water are added in reaction kettle and reacted, is filtered
After obtain complex liquid;After obtained complex liquid is mixed with obtained enzymolysis liquid, converted starch is then added, it is spraying by homogeneous
It dries to get the composite flavored extract is arrived.The composite flavored extract of invention preparation gives off a strong fragrance, and fragrance is held
Long, it can effectively inhibit the growth of microorganism, remove raw meat, smell of having a strong smell, can still keep good fragrance at high temperature.This hair
It is bright to belong to technical field of food additives, and in particular to a kind of preparation method of composite flavored extract.
In view of the deficiencies of the prior art, the object of the present invention is to provide a kind of preparation method of composite flavored extract,
Not only flavouring effect is good for the composite flavored extract prepared, and fragrance is lasting, and can effectively inhibit the growth of microorganism,
Prevent food spoilage.
The present invention provides a kind of preparation methods of composite flavored extract, specifically includes the following steps:
(1), fresh bonito stick is chosen, carries out break process after cleaning, it is spare;Choose ginger powder, garlic powder, fennel seeds, mulberry leaf,
Ground Cloves, salt and water are spare;
(2), tissue mashing machine is added in the bonito stick after break process in step (1), water is added, is smashed to pieces, at mashing
Reason, obtains bonito stick slurries;
(3), the bonito stick slurries for obtaining step (2) adjust pH to 5-7, bromelain are added under the conditions of 40-60 DEG C
Enzyme, enzymatic hydrolysis obtained enzymolysis product after 1-2 hours;
(4), the enzymolysis product that step (3) obtains is heated to 80-100 DEG C of progress destroy the enzyme treatment 10-30min, be centrifuged
Enzymolysis liquid is obtained after filter;
(5), reaction ax is added in ginger powder, garlic powder, fennel seeds, mulberry leaf, Ground Cloves, salt and the water chosen step (1)
In, it is warming up to 80-100 DEG C after mixing evenly, is cooled to room temperature after being stirred to react 1-2h, complex liquid is obtained after filtering;
(6), after mixing the complex liquid that step (5) obtains with the enzymolysis liquid that step (4) obtains, converted starch is added, stirs
Enter homogenizer after mixing uniformly, obtains homogenizing fluid;
(7), homogenizing fluid obtained in step (6) is arrived into the composite flavored extract by spray drying.
The mass ratio of the bonito stick and water after break process in the step (2) is 1:3-6.
The quality for the bromelain being added in the step (3) is the 0.5-1.5% of bonito stick grind slurries quality.
Ginger powder, garlic powder, fennel seeds, mulberry leaf, Ground Cloves, salt and water in the step (5), by weight
Are as follows: 20-40 parts of ginger powder, 10-30 parts of garlic powder, 10-30 parts of fennel seeds, 5-15 parts of mulberry leaf, 10-30 parts of Ground Cloves, salt 10-20
Part and water 100-150 parts.
The mass ratio of enzymolysis liquid, complex liquid and converted starch in the step (6) is 1:4-6:1.
Spray drying condition in the step (7) are as follows: inlet temperature is 180-200 DEG C, and outlet temperature is 80-100 DEG C.
Preferably, the quality of the bonito stick after the break process in the step (2) and water is 1:4-5.
Preferably, the quality for the bromelain being added in the step (3) is the 0.8-1.2% of bonito stick grind slurries quality.
Preferably, ginger powder, garlic powder, fennel seeds, mulberry leaf, Ground Cloves, salt and the water in the step (5), by weight
Part meter are as follows: 25-35 parts of ginger powder, 15-25 parts of garlic powder, 15-25 parts of fennel seeds, 8-12 parts of mulberry leaf, 15-25 parts of Ground Cloves, food
13-17 parts and water 110-130 parts of salt.
The mass ratio of enzymolysis liquid, complex liquid and converted starch in the step (6) is 1:4.5-5.5:1.
The activity of the bromelain is 500,000 U/g.
The invention has the benefit that
(1) the composite flavored extract of preparation method provided by the invention, preparation gives off a strong fragrance, and fragrance is lasting.
(2) composite flavored extract prepared by the present invention can effectively inhibit the growth of microorganism, prevent food spoilage.
(3) the composite flavored extract high-temperature resistant result that the present invention prepares is good, can still keep at high temperature good
Good fragrance value.
(4) composite flavored extract prepared by the present invention can also effectively dispel fishy smell and smell of mutton.
Specific embodiment
A kind of preparation method of the composite flavored extract of embodiment 1
(1), fresh bonito stick is chosen, carries out break process after cleaning, it is spare;Choose ginger powder, garlic powder, fennel seeds, mulberry leaf,
Ground Cloves, salt and water are spare;
(2), tissue mashing machine is added in the bonito stick after break process in step (1), water is added, is smashed to pieces, at mashing
Reason, obtains bonito stick slurries, wherein the mass ratio of bonito stick and water after break process is 1:3;
(3), the bonito stick slurries for obtaining step (2) adjust pH to 5, bromelain are added under the conditions of 40 DEG C, digest
After 1 hour, enzymolysis product is obtained, the quality of the bromelain of addition is the 0.5% of bonito stick grind slurries quality;
(4), the enzymolysis product that step (3) obtains is heated to 80 DEG C of progress destroy the enzyme treatment 10min, is obtained after centrifugal filtration
Enzymolysis liquid;
(5), by 10 parts of 20 parts of ginger powder, the garlic powder by weight of step (1) selection, 10 parts of fennel seeds, mulberry leaf 5
In part, 10 parts of Ground Cloves, 10 parts of salt and 100 parts of addition reaction kettles of water, it is warming up to 80 DEG C, after being stirred to react 1h after mixing evenly
It is cooled to room temperature, complex liquid is obtained after filtering;
(6), after mixing the complex liquid that step (5) obtains with the enzymolysis liquid that step (4) obtains, denaturation is then added and forms sediment
Powder enters homogenizer after mixing evenly, obtains homogenizing fluid, wherein the mass ratio of enzymolysis liquid, complex liquid and converted starch is 1:4:
1;
(7), by the homogenizing fluid of obtaining in step (6) by spray drying, spray drying condition are as follows: inlet temperature is
180 DEG C, outlet temperature be 80 DEG C to get arrive the composite flavored extract.
A kind of preparation method of the composite flavored extract of embodiment 2
(1), fresh bonito stick is chosen, carries out break process after cleaning, it is spare;Choose ginger powder, garlic powder, fennel seeds, mulberry leaf,
Ground Cloves, salt and water are spare;
(2), tissue mashing machine is added in the bonito stick after break process in step (1), water is added, is smashed to pieces, at mashing
Reason, obtains bonito stick slurries, wherein the mass ratio of bonito stick and water after break process is 1:6;
(3), the bonito stick slurries for obtaining step (2) adjust pH to 7, bromelain are added under the conditions of 60 DEG C, digest
After 2 hours, enzymolysis product is obtained, the quality of the bromelain of addition is the 1.5% of bonito stick grind slurries quality;
(4), the enzymolysis product that step (3) obtains is heated to 100 DEG C of progress destroy the enzyme treatment 30min, after centrifugal filtration
To enzymolysis liquid;
(5), by 30 parts of 40 parts of ginger powder, the garlic powder by weight of step (1) selection, 30 parts of fennel seeds, mulberry leaf 15
Part, 30 parts of Ground Cloves, 20 parts of salt and 150 parts of water are added in reaction kettle, are warming up to 100 DEG C after mixing evenly, are stirred to react 2h
After be cooled to room temperature, complex liquid is obtained after filtering;
(6), after mixing the complex liquid that step (5) obtains with the enzymolysis liquid that step (4) obtains, denaturation is then added and forms sediment
Powder enters homogenizer after mixing evenly, obtains homogenizing fluid, wherein the mass ratio of enzymolysis liquid, complex liquid and converted starch is 1:6:
1;
(7), by the homogenizing fluid of obtaining in step (6) by spray drying, spray drying condition are as follows: inlet temperature is
200 DEG C, outlet temperature be 100 DEG C to get arrive the composite flavored extract.
A kind of preparation method of the composite flavored extract of embodiment 3
(1), fresh bonito stick is chosen, carries out break process after cleaning, it is spare;Choose ginger powder, garlic powder, fennel seeds, mulberry leaf,
Ground Cloves, salt and water are spare;
(2), tissue mashing machine is added in the bonito stick after break process in step (1), water is added, is smashed to pieces, at mashing
Reason, obtains bonito stick slurries, wherein the quality of bonito stick and water after break process is 1:4;
(3), the bonito stick slurries for obtaining step (2) adjust pH to 6, bromelain are added under the conditions of 50 DEG C, digest
After 1.5 hours, enzymolysis product is obtained, the quality of the bromelain of addition is the 0.8% of bonito stick grind slurries quality;
(4), the enzymolysis product that step (3) obtains is heated to 90 DEG C of progress destroy the enzyme treatment 20min, is obtained after centrifugal filtration
Enzymolysis liquid;
(5), by 15 parts of 25 parts of ginger powder, the garlic powder by weight of step (1) selection, 15 parts of fennel seeds, mulberry leaf 8
In part, 15 parts of Ground Cloves, 13 parts of salt and 110 parts of addition reaction kettles of water, it is warming up to 90 DEG C after mixing evenly, is stirred to react 1.5h
After be cooled to room temperature, complex liquid is obtained after filtering;
(6), after mixing the complex liquid that step (5) obtains with the enzymolysis liquid that step (4) obtains, denaturation is then added and forms sediment
Powder enters homogenizer after mixing evenly, obtains homogenizing fluid, wherein the mass ratio of enzymolysis liquid, complex liquid and converted starch is 1:
4.5:1;
(7), by the homogenizing fluid of obtaining in step (6) by spray drying, spray drying condition are as follows: inlet temperature is
200 DEG C, outlet temperature be 100 DEG C to get arrive the composite flavored extract.
A kind of preparation method of the composite flavored extract of embodiment 4
(1), fresh bonito stick is chosen, carries out break process after cleaning, it is spare;Choose ginger powder, garlic powder, fennel seeds, mulberry leaf,
Ground Cloves, salt and water are spare;
(2), tissue mashing machine is added in the bonito stick after break process in step (1), water is added, is smashed to pieces, at mashing
Reason, obtains bonito stick slurries, wherein the quality of bonito stick and water after break process is 1:5;
(3), the bonito stick slurries for obtaining step (2) adjust pH to 6, bromelain are added under the conditions of 50 DEG C, digest
After 1.5 hours, enzymolysis product is obtained, the quality of the bromelain of addition is the 1.2% of bonito stick grind slurries quality;
(4), the enzymolysis product that step (3) obtains is heated to 90 DEG C of progress destroy the enzyme treatment 20min, is obtained after centrifugal filtration
Enzymolysis liquid;
(5), by 25 parts of 35 parts of ginger powder, the garlic powder by weight of step (1) selection, 25 parts of fennel seeds, mulberry leaf 12
In part, 25 parts of Ground Cloves, 17 parts of salt and 130 parts of addition reaction kettles of water, it is warming up to 90 DEG C after mixing evenly, is stirred to react 1.5h
After be cooled to room temperature, complex liquid is obtained after filtering;
(6), after mixing the complex liquid that step (5) obtains with the enzymolysis liquid that step (4) obtains, denaturation is then added and forms sediment
Powder enters homogenizer after mixing evenly, obtains homogenizing fluid, wherein the mass ratio of enzymolysis liquid, complex liquid and converted starch is 1:
5.5:1;
(7), by the homogenizing fluid of obtaining in step (6) by spray drying, spray drying condition are as follows: inlet temperature is
180 DEG C, outlet temperature be 80 DEG C to get arrive the composite flavored extract.
A kind of preparation method of the composite flavored extract of embodiment 5
(1) fresh bonito stick is chosen, carries out break process after cleaning, it is spare;Choose ginger powder, garlic powder, fennel seeds, mulberry leaf,
Ground Cloves, salt and water are spare;
(2) tissue mashing machine is added in the bonito stick after break process in step (1), water is added, is smashed to pieces, at mashing
Reason, obtains bonito stick slurries, wherein the quality of bonito stick and water after break process is 1:4.5;
(3) the bonito stick slurries for obtaining step (2) adjust pH to 6, and bromelain, enzymatic hydrolysis 2 are added under the conditions of 50 DEG C
After hour, enzymolysis product is obtained, the quality of the bromelain of addition is the 1% of bonito stick grind slurries quality;
(4) enzymolysis product that step (3) obtains is heated to 90 DEG C of progress destroy the enzyme treatment 20min, is obtained after centrifugal filtration
Enzymolysis liquid;
(5) by 20 parts of 30 parts of ginger powder, the garlic powder by weight of step (1) selection, 20 parts of fennel seeds, mulberry leaf 10
In part, 20 parts of Ground Cloves, 15 parts of salt and 120 parts of addition reaction kettles of water, it is warming up to 100 DEG C after mixing evenly, is stirred to react 2h
After be cooled to room temperature, complex liquid is obtained after filtering;
(6) after mixing the complex liquid that step (5) obtains with the enzymolysis liquid that step (4) obtains, converted starch is then added,
Enter homogenizer after mixing evenly, obtain homogenizing fluid, wherein the mass ratio of enzymolysis liquid, complex liquid and converted starch is 1:5:1;
(7) by the obtained homogenizing fluid in step (6) by spray drying, spray drying condition are as follows: inlet temperature 190
DEG C, outlet temperature be 90 DEG C to get arrive the composite flavored extract.
A kind of preparation method of the composite flavored extract of comparative example 1
Preparation method substantially with embodiment 5, with embodiment 5 the difference is that:
Step is added without mulberry leaf in (5).
A kind of preparation method of the composite flavored extract of comparative example 2
Preparation method substantially with embodiment 5, with embodiment 5 the difference is that:
Enzymolysis time in step (3) is 0.5h.
A kind of preparation method of the composite flavored extract of comparative example 3
Preparation method substantially with embodiment 5, with embodiment 5 the difference is that:
Enzymolysis time in step (3) is 2.5h.
A kind of preparation method of the composite flavored extract of comparative example 4
Preparation method substantially with embodiment 5, with embodiment 5 the difference is that:
The mass ratio of enzymolysis liquid, complex liquid and converted starch in step (6) is 1:3:1.
A kind of preparation method of the composite flavored extract of comparative example 5
Preparation method substantially with embodiment 5, with embodiment 5 the difference is that:
The mass ratio of enzymolysis liquid, complex liquid and converted starch in step (6) is 1:7:1.
The seasoning that 6 Chinese patent of comparative example, 201410209287.9 embodiment 1 provides.
1 bacteriostasis research of experimental example
The seasoning point that composite flavored extract and comparative example 6 prepared by 1-5 of the embodiment of the present invention, comparative example 1 provides
It is not mixed with dehydrated alcohol according to the ratio of 1:1 (g:mL), different samples to be tested is prepared.Take 25g naturally rotten at room temperature
The octopus lost mixes with 200mL sterile saline after chopping, fullys shake, and 1mL bacterium solution is taken to be inoculated into beef extract-peptone liquid
In body culture medium, it is enriched with nature spoilage organisms for 24 hours in 37 DEG C of constant temperature incubations.The liquid training for taking 100 μ L to contain spoilage organisms with liquid-transfering gun
Then plus 15mL bactericidal nurishing agar medium it supports based in sterilizes culture dish, is uniformly mixed, culture medium is put into before not solidifying to go out
Bacterium Oxford cup, takes out well after culture medium is cooling, and every hole is added 60 μ L of sample liquid, then cultivates at 37 DEG C for 24 hours, each
Sample is averaged after measuring antibacterial circle diameter 10 times, as a result as shown in table 1 below:
Table 1
Antibacterial circle diameter (mm) | |
Embodiment 1 | 35.2 |
Embodiment 2 | 34.6 |
Embodiment 3 | 35.4 |
Embodiment 4 | 35.6 |
Embodiment 5 | 36.5 |
Comparative example 1 | 28.2 |
Comparative example 6 | 26.2 |
By above-mentioned experimental result it is found that the composite flavored extract provided in 1-5 of the embodiment of the present invention has antibacterial work
With can inhibit the growth of the spoilage organisms in octopus, be conducive to the preservation of octopus, mulberry leaf, fungistatic effect are free of in comparative example 1
It is substantially reduced, illustrates that mulberry leaf are inhibited to the spoilage organisms in octopus.
2 Evaluation of Thermal Stability of experimental example
By embodiment 1-5 and comparative example 2-3 the composite flavored extract being prepared use respectively 0.2% saline at
Concentration is the sample solution of 0.5g/mL, takes different sample solution 100mL in 250mL reagent bottle respectively, and sealing bottleneck prevents
Moisture drain evaporation is put into 2h in 100 DEG C of water-bath, takes out beaker, is cooled to room temperature, using quantitative description method to compound increasing
Odor type extract carries out sensory evaluation, forms evaluation group by 20 sensory evaluation persons after sensory evaluation and screening, right
The organoleptic indicator of each solution carries out evaluation marking, collects evaluation result, is averaged, for statistical analysis.Composite flavored mentions
Taking the sensory evaluation of object to score, scale is as shown in table 2, and aroma change is indicated using aroma change rate:
R=(A0-A1)/A0× 100%
Wherein: after R- sample solution water-bath 2h compared with when not heating aroma strength change rate;
A1Aroma strength after sample solution water-bath 2h;
A0Aroma strength when sample solution does not heat.
Table 2
Scoring | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 |
Aroma strength | It is extremely weak | It is very weak | It is weaker | It is slightly weak | It is medium | It is slightly strong | It is relatively strong | It is very strong | It is extremely strong |
Table 3
Change rate (%) | |
Embodiment 1 | 12.5 |
Embodiment 2 | 13.6 |
Embodiment 3 | 11.9 |
Embodiment 4 | 12.4 |
Embodiment 5 | 11.5 |
Comparative example 2 | 16.9 |
Comparative example 3 | 20.2 |
By the experimental result of table 3 it is found that the high temperature resistance for the composite flavored extract that 1-5 of the embodiment of the present invention is provided
Good, wherein the aroma change rate in embodiment 5 is 11.5%, is lower than comparative example 2-3.
The removal evaluation of 3 smell of mutton of experimental example
The composite flavored that embodiment 1-5 and comparative example the 4-5 preparation of equivalent are added during cooking mutton extracts
Object, control group are to be added without composite flavored extract, are then had a strong smell again by each composite flavored extract removal of sensory evaluation
The effect of taste, sensory evaluation scores, and scale is as shown in table 4, and statistic analysis result is as shown in table 5:
Table 4
Scoring | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 |
Smell of mutton intensity | It is extremely weak | It is very weak | It is weaker | It is slightly weak | It is medium | It is slightly strong | It is relatively strong | It is very strong | It is extremely strong |
Table 5
Smell of mutton | |
Control group | 8.2 |
Embodiment 1 | 2.2 |
Embodiment 2 | 1.9 |
Embodiment 3 | 1.6 |
Embodiment 4 | 1.3 |
Embodiment 5 | 2.1 |
Comparative example 4 | 4.1 |
Comparative example 5 | 3.3 |
By the experimental result of table 3 it is found that the composite flavored extract that 1-5 of the embodiment of the present invention is provided can effectively be gone
Except the smell of mutton in mutton, it is better than comparative example 4-5.
Only as described above, is only a preferred embodiment of the present invention, when the model that cannot limit implementation of the invention with this
It encloses, i.e., generally according to simple equivalent changes and modifications made by content documented by the claims in the present invention and description of the invention,
All it is still covered by the claims of the invention.It is searched in addition, abstract part and title are intended merely to auxiliary patent document
It seeks and being used, the interest field being not intended to limit the invention.
Claims (10)
1. a kind of extracting method of bromelain, it is characterised in that: the following steps are included:
1) it pre-processes: fresh pineapple or pineapple stem being cleaned, precooling treatment, crushed, juicing;
2) it digests: distilled water, lignoenzyme and stabilizer being added in the pineapple juice with pulp, enzyme deactivation after enzymatic hydrolysis adds
Enter cellulase and pectase, digests, enzyme deactivation after enzymatic hydrolysis, filtering;
3) isoelectric precipitation: adjusting filtrate pH value, stirs, and stands, filtering;
4) distilled water, ultrafiltration ultrafiltration: are added in precipitating;
5) kept dry: stagnant liquid concentrated by rotary evaporation being freeze-dried, grinding, and stabilizer is added, and stirring obtains bromelain.
2. the extracting method of bromelain as described in claim 1, it is characterised in that: precooling treatment in the step 1)
Temperature be 0~2 DEG C, pre-coo time be 21~25h.
3. the extracting method of bromelain as described in claim 1, it is characterised in that: have pulp in the step 2)
Pineapple juice and distilled water mass ratio be 1:6~10.
4. a kind of extracting method of bromelain as described in claim 1, it is characterised in that: wooden in the step 2)
Plain enzyme additional amount is that 0.2~0.4%, pH is 3.5~4.2, and temperature is 38~43 DEG C, and enzymolysis time is 3~3.4h;Cellulase
Additional amount with pectase is respectively 0.3~0.5% and 0.1~0.2%, and hydrolysis temperature is 53~56 DEG C, and pH is 2.7~3.2,
Enzymolysis time is 2.0~2.3h.
5. a kind of preparation method of composite flavored extract, it is characterised in that: specifically includes the following steps:
(1), fresh bonito stick is chosen, carries out break process after cleaning, it is spare;Choose ginger powder, garlic powder, fennel seeds, mulberry leaf, cloves
Powder, salt and water are spare;
(2), tissue mashing machine is added in the bonito stick after break process in step (1), water is added, smashed to pieces, mashing processing, obtained
To bonito stick slurries;
(3), the bonito stick slurries for obtaining step (2) adjust pH to 5-7, bromelain, enzyme are added under the conditions of 40-60 DEG C
Solution obtained enzymolysis product after 1-2 hours;
(4), the enzymolysis product that step (3) obtains is heated to 80-100 DEG C of progress destroy the enzyme treatment 10-30min, after centrifugal filtration
Obtain enzymolysis liquid;
(5), ginger powder, garlic powder, fennel seeds, mulberry leaf, Ground Cloves, salt and the water chosen step (1) are added in reaction kettle, stir
It is warming up to 80-100 DEG C after mixing uniformly, is cooled to room temperature after being stirred to react 1-2h, complex liquid is obtained after filtering;
(6), after mixing the complex liquid that step (5) obtains with the enzymolysis liquid that step (4) obtains, converted starch is then added, stirs
Enter homogenizer after mixing uniformly, obtains homogenizing fluid;
(7), the obtained homogenizing fluid in step (6) is arrived into the composite flavored extract by spray drying.
6. preparation method as claimed in claim 5, it is characterised in that: bonito stick after break process in the step (2) and
The mass ratio of water is 1:3-6;Preferably, the quality of the bonito stick after the break process in the step (2) and water is 1:4-5.
7. preparation method as claimed in claim 5, it is characterised in that: the matter for the bromelain being added in the step (3)
Amount is the 0.5-1.5% of bonito stick grind slurries quality;Preferably, the quality for the bromelain being added in the step (3) is bonito stick
The 0.8-1.2% of grind slurries quality.
8. preparation method as claimed in claim 5, it is characterised in that: ginger powder, garlic powder, fennel seeds in the step (5),
Mulberry leaf, Ground Cloves, salt and water, by weight are as follows: 20-40 parts of ginger powder, 10-30 parts of garlic powder, 10-30 parts of fennel seeds,
5-15 parts of mulberry leaf, 10-30 parts of Ground Cloves, 10-20 parts of salt and 100-150 parts of water;Preferably, the ginger powder in the step (5),
Garlic powder, fennel seeds, mulberry leaf, Ground Cloves, salt and water, by weight are as follows: 25-35 parts of ginger powder, 15-25 parts of garlic powder,
15-25 parts of fennel seeds, 8-12 parts of mulberry leaf, 15-25 parts of Ground Cloves, 13-17 parts of salt and 110-130 parts of water.
9. preparation method as claimed in claim 5, it is characterised in that: enzymolysis liquid, complex liquid and denaturation in the step (6)
The mass ratio of starch is 1:4-6:1;Preferably, the mass ratio of the enzymolysis liquid in the step (6), complex liquid and converted starch is
1:4.5-5.5:1。
10. preparation method as claimed in claim 5, it is characterised in that: the spray drying condition in the step (7) are as follows: into
Mouth temperature is 180-200 DEG C, and outlet temperature is 80-100 DEG C.
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