CN108641804A - A kind of method of ultrasonic wave added aqueous enzymatic extraction setose abelmoschus seed oil and protein - Google Patents
A kind of method of ultrasonic wave added aqueous enzymatic extraction setose abelmoschus seed oil and protein Download PDFInfo
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- CN108641804A CN108641804A CN201810331359.5A CN201810331359A CN108641804A CN 108641804 A CN108641804 A CN 108641804A CN 201810331359 A CN201810331359 A CN 201810331359A CN 108641804 A CN108641804 A CN 108641804A
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- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
- C11B1/02—Pretreatment
- C11B1/025—Pretreatment by enzymes or microorganisms, living or dead
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D21/00—Separation of suspended solid particles from liquids by sedimentation
- B01D21/26—Separation of sediment aided by centrifugal force or centripetal force
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D61/00—Processes of separation using semi-permeable membranes, e.g. dialysis, osmosis or ultrafiltration; Apparatus, accessories or auxiliary operations specially adapted therefor
- B01D61/14—Ultrafiltration; Microfiltration
- B01D61/145—Ultrafiltration
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- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
- C11B1/10—Production of fats or fatty oils from raw materials by extracting
- C11B1/106—Production of fats or fatty oils from raw materials by extracting using ultra-sounds
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Abstract
The invention discloses a kind of methods of ultrasonic wave added aqueous enzymatic extraction setose abelmoschus seed oil and protein, setose abelmoschus seed powder are mixed with complex enzyme, the ultrasonic wave added enzyme digestion reaction at 30~60 DEG C, 100~200W of ultrasonic power；Enzymolysis liquid low-speed centrifugal obtains free oil I, emulsion I, protein liquid and waste residue；Protein liquid ultrafiltration obtains setose abelmoschus seed protein；I high speed centrifugation of emulsion obtains free oil II and emulsion II；Emulsion II is mixed with ethyl alcohol, ultrasonic wave added demulsification, is then centrifuged for obtaining demulsification oil；Free oil I, free oil II and demulsification oil are merged and obtain setose abelmoschus seed oil.The present invention solves the problems such as traditional aqueous enzymatic method free oil recovery rate is low, and residual oil content height and protein comprehensively utilize deficiency in emulsion.
Description
Technical field
The invention belongs to agricultural and sideline product retrofit and its byproduct comprehensive utilization technical fields, and in particular to a kind of ultrasound
The method for assisting aqueous enzymatic extraction setose abelmoschus seed oil and protein.
Background technology
Okra (Abelmoschus esculentus (Linn.) Moench) alias kidney tonifying grass, gumbo, swordweed etc.,
For Malvaceae (Malvaceae) Abelmoschus (Abelmoschus Medic) annual herb plant.Grease accounts for setose abelmoschus seed quality
15%~19%, contain about 40% linoleic acid in setose abelmoschus seed grease, and saturated fatty acid, monounsaturated fatty acids and more
The ratio of unsaturated fatty acid is more balanced, is a kind of higher vegetable oil of nutritive value.The study found that setose abelmoschus seed oil can change
Kind lipid-metabolism, enhancing development, adjusts immune system, anticancer, anti-aging, promotes brain and regard prevention of cardiovascular disease
The development etc. of power.In addition, also containing protein 20% in setose abelmoschus seed, content reaches 55% after degreasing, is rich in 17 kinds of amino acid,
Including 7 kinds of amino acid needed by human, are a kind of good protein resources, therefore using setose abelmoschus seed oil compound protein, answer
For fields such as functional food and health medicines.
Currently, the preparation to setose abelmoschus seed oil takes traditional extraction method and squeezing method more.Though solvent extraction method has
The advantage that residual oil content is low, protein denaturation is small, dissolvable agents residual, the disadvantage of safety difference are also especially prominent, are not suitable for Huang Qiu
Seed oil of sunflower；And rule not only low production efficiency but also Oxidation of Fat and Oils is serious is squeezed, it is not suitable for setose abelmoschus seed oil yet.
Aqueous enzymatic method is a kind of in recent years emerging extract oil technique, and with machinery and enzymolysis being main means keeps plant cell wall broken
It is broken, what the complexs such as lipoprotein and lipopolysaccharides in oil plant cell of being degraded by enzyme were formed to destroy oil plant during defibrination etc.
It is wrapped in the lipoprotein membrane of oil droplets, to discharge grease, improves free oil extracted amount.Aqueous enzymatic extraction oil technique quilt earliest
For coconut oil, be then gradually available for again setose abelmoschus seed oil, soybean oil, peanut oil, rapeseed oil, cottonseed oil, sunflower oil, can
It can oil and palm oil etc..2016, Chen Xuan etc. prepared setose abelmoschus seed oil technique and its oxidation-stabilized Journal of Sex Research using aqueous enzymatic method, tied
Fruit shows that oil yield is 14.25%, and setose abelmoschus seed oil shelf life prepared by aqueous enzymatic method is 230 days, shorter than extraction legal system
Standby 281 days are longer than 164 days of squeezing method preparation.Aqueous enzymatic method can propose simultaneously in a mild condition grease in oilseeds and
Protein largely avoids the destruction to grease and protein.Therefore, compared with conventional method, aqueous enzymatic extraction
Oil quality is more preferable, protein denaturation degree smaller, the more transparent clarification of color and luster.Aqueous enzymatic method is carried out due to digesting in water phase, phosphorus
Fat enters water phase, because of the step of without degumming process, can not only reducing chemical refining, moreover it is possible to improve the quality of oil.But mesh
The process that preceding aqueous enzymatic method prepares setose abelmoschus seed oil can form one layer of emulsion, and the emulsion formed cannot be broken well
It removes so that the yield of free oil reduces, and is applied to industrialized production to greatly limit aqueous enzymatic method.Therefore, exploitation is a kind of suitable
For industrialized production, and there can be important meaning simultaneously in the method that higher extracted rate extracts setose abelmoschus seed oil and protein
Justice.
Invention content
The present invention provides a kind of methods of ultrasonic wave added aqueous enzymatic extraction setose abelmoschus seed oil and protein, solve tradition
Aqueous enzymatic method free oil recovery rate is low, and residual oil content is high in emulsion and protein comprehensively utilizes the problems such as insufficient.
The technical solution adopted by the present invention to solve the technical problems is：
A kind of method of ultrasonic wave added aqueous enzymatic extraction setose abelmoschus seed oil and protein, including：
1) setose abelmoschus seed powder and water are pressed 1:4~8 solid-liquid ratio (w/v, g/mL) mixing, adjust pH value be 7.0~
9.0, the complex enzyme for being equivalent to setose abelmoschus seed powder quality 1~5% is added, the complex enzyme includes cellulase, hemicellulose
At least one of plain enzyme, pectase, zytase further include protease, and in complex enzyme protease content be 20~
45% (mass percent)；Then under conditions of 30~60 DEG C of temperature, 100~200W of ultrasonic power, ultrasonic wave added enzymolysis is anti-
60~240min is answered, enzymolysis liquid is obtained；
2) the enzymolysis liquid low-speed centrifugal for obtaining step 1), the free oil I being distributed from top to bottom, emulsion I, albumen
Liquid and waste residue；
3) the protein liquid ultrafiltration that step 2) obtains is taken, trapped fluid concentration, drying, as setose abelmoschus seed protein are taken；
4) I high speed centrifugation of emulsion that step 2) obtains is taken, the breast positioned at the free oil II on upper layer and positioned at lower layer is obtained
Shape liquid II；
5) ethyl alcohol of the emulsion II for obtaining step 4) and volumetric concentration 20~60% by volume 1:1~3 ratio
Mixing, under conditions of 100~150W of ultrasonic power, ultrasonic wave added 0.5~1.5h of demulsification, then medium-speed centrifuge, is demulsified
Oil；The free oil II and the demulsification oil that free oil I that step 2) obtains, step 4) are obtained merge, as setose abelmoschus seed oil.
In one embodiment：In the step 1), by setose abelmoschus seed screening, clean, 50~75 DEG C of 22~26h of forced air drying,
High speed smashes it through 40~60 mesh sieve, obtains the setose abelmoschus seed powder.
In one embodiment：In the step 1), complex enzyme includes that mass ratio is 1~3:0.5~1.5:0.5~1.5 fibre
The plain enzyme of dimension, pectase and protease.
In one embodiment：In the step 2), low-speed centrifugal is to centrifuge 10~20min in 2000~3000r/min.
In one embodiment：In the step 3), uses molecular cut off to carry out ultrafiltration for the ultrafiltration membrane of 10~30kDa, take
Trapped fluid is concentrated under reduced pressure, and is freeze-dried 45~50h, obtains the setose abelmoschus seed protein.
In one embodiment：In the step 4), high speed centrifugation is to centrifuge 10~15min in 10000~15000r/min.
In one embodiment：In the step 5), medium-speed centrifuge is to centrifuge 10~15min in 5000~8000r/min.
Compared with the background art, it has the following advantages that the technical program：
The present invention accelerates grease using the effects that the cavitation effect of ultrasonic wave, fuel factor, mechanical effect and chemical effect
Reaction rate, promote the precipitation of autumn seed oil of sunflower, relatively traditional aqueous enzymatic method technique makes the recovery rate of free oil significantly improve,
And oil quality is high；Ultrasonic wave can form microvesicle in liquid medium, and microvesicle can discharge more energy in rupture process, can
It is demulsified to obtain reaction rate with improving ethyl alcohol；The purity higher of setose abelmoschus seed protein through hyperfiltration treatment, promotes the comprehensive of protein
It closes and utilizes；Digesting needed for preparing during setose abelmoschus seed oil and protein are implemented, ultrasound, centrifugation, ultrafiltration, freeze-drying etc.
Equipment and technology maturity is high, feasibility is strong, is suitable for large-scale industrial production.
Description of the drawings
The invention will be further described with reference to the accompanying drawings and examples.
Fig. 1 is the process route signal of the ultrasonic wave added aqueous enzymatic extraction setose abelmoschus seed oil of the present invention and the method for protein
Figure.
Specific implementation mode
Present disclosure is illustrated below by embodiment：
Embodiment 1
1) setose abelmoschus seed screened successively, cleaned, 50 DEG C of forced air dryings for 24 hours, using medicinal herb grinder high speed crush,
40 mesh sieve is crossed, the setose abelmoschus seed powder is obtained；Setose abelmoschus seed powder and water are pressed 1:The solid-liquid ratio of 4 (w/v) mixes, and adjusts pH
Value is 7.5, adds the complex enzyme for being equivalent to setose abelmoschus seed powder quality 1%, and the complex enzyme includes that mass ratio is 2:1:1
Cellulase, pectase and protease；Then under conditions of temperature 50 C, ultrasonic power 200W, ultrasonic wave added enzyme digestion reaction
60min obtains enzymolysis liquid；
2) enzymolysis liquid for obtaining step 1) 2500r/min centrifuge 15min, the free oil I being distributed from top to bottom,
Emulsion I, protein liquid and waste residue；
3) after taking the protein liquid suitably dilution that step 2) obtains, molecular cut off is used to be surpassed for the ultrafiltration membrane of 10kDa
Filter, takes trapped fluid to be concentrated under reduced pressure, and is freeze-dried 48h, as setose abelmoschus seed protein；
4) it takes the emulsion I that step 2) obtains to centrifuge 10min in 10000r/min, obtains II He of free oil positioned at upper layer
Positioned at the emulsion II of lower layer；
5) ethyl alcohol of the emulsion II for obtaining step 4) and volumetric concentration 20% by volume 1:1 ratio mixing,
Under conditions of ultrasonic power 150W, then ultrasonic wave added demulsification 0.5h centrifuges 10min in 5000r/min, obtains positioned at upper layer
Demulsification oil；The free oil II and the demulsification oil that free oil I that step 2) obtains, step 4) are obtained merge, as setose abelmoschus seed
Oil.
Among the present embodiment, free oil recovery rate is 83%, percent of total oil 91%；Protein recovery is 87%,
Setose abelmoschus seed lipidated protein is 85%.The rate of abolishing of ultrasonic wave added ethyl alcohol demulsification method emulsion may be up to 91~92%.
Embodiment 2
1) setose abelmoschus seed screened successively, cleaned, 60 DEG C of forced air dryings for 24 hours, using medicinal herb grinder high speed crush,
40 mesh sieve is crossed, the setose abelmoschus seed powder is obtained；Setose abelmoschus seed powder and water are pressed 1:The solid-liquid ratio of 5 (w/v) mixes, and adjusts pH
Value is 8.0, adds the complex enzyme for being equivalent to setose abelmoschus seed powder quality 1%, and the complex enzyme includes that mass ratio is 2:1:1
Cellulase, pectase and protease；Then under conditions of temperature 60 C, ultrasonic power 200W, ultrasonic wave added enzyme digestion reaction
120min obtains enzymolysis liquid；
2) enzymolysis liquid for obtaining step 1) 2500r/min centrifuge 15min, the free oil I being distributed from top to bottom,
Emulsion I, protein liquid and waste residue；
3) after taking the protein liquid suitably dilution that step 2) obtains, molecular cut off is used to be surpassed for the ultrafiltration membrane of 10kDa
Filter, takes trapped fluid to be concentrated under reduced pressure, and is freeze-dried 48h, as setose abelmoschus seed protein；
4) it takes the emulsion I that step 2) obtains to centrifuge 10min in 10000r/min, obtains II He of free oil positioned at upper layer
Positioned at the emulsion II of lower layer；
5) ethyl alcohol of the emulsion II for obtaining step 4) and volumetric concentration 20% by volume 1:1 ratio mixing,
Under conditions of ultrasonic power 150W, then ultrasonic wave added demulsification 0.5h centrifuges 10min in 5000r/min, obtains positioned at upper layer
Demulsification oil；The free oil II and the demulsification oil that free oil I that step 2) obtains, step 4) are obtained merge, as setose abelmoschus seed
Oil.
Among the present embodiment, free oil recovery rate is 82%, percent of total oil 90.8%；Protein recovery is
85%, setose abelmoschus seed lipidated protein is 87%.The rate of abolishing of ultrasonic wave added ethyl alcohol demulsification method emulsion may be up to 91~92%.
Embodiment 3
1) setose abelmoschus seed screened successively, cleaned, 50 DEG C of forced air dryings for 24 hours, using medicinal herb grinder high speed crush,
40 mesh sieve is crossed, the setose abelmoschus seed powder is obtained；Setose abelmoschus seed powder and water are pressed 1:The solid-liquid ratio of 4 (w/v) mixes, and adjusts pH
Value is 7.5, adds the complex enzyme for being equivalent to setose abelmoschus seed powder quality 1%, and the complex enzyme includes that mass ratio is 2:1:1
Cellulase, pectase and protease；Then under conditions of 40 DEG C of temperature, ultrasonic power 100W, ultrasonic wave added enzyme digestion reaction
240min obtains enzymolysis liquid；
2) enzymolysis liquid for obtaining step 1) 2500r/min centrifuge 15min, the free oil I being distributed from top to bottom,
Emulsion I, protein liquid and waste residue；
3) after taking the protein liquid suitably dilution that step 2) obtains, molecular cut off is used to be surpassed for the ultrafiltration membrane of 10kDa
Filter, takes trapped fluid to be concentrated under reduced pressure, and is freeze-dried 48h, as setose abelmoschus seed protein；
4) it takes the emulsion I that step 2) obtains to centrifuge 10min in 10000r/min, obtains II He of free oil positioned at upper layer
Positioned at the emulsion II of lower layer；
5) ethyl alcohol of the emulsion II for obtaining step 4) and volumetric concentration 20% by volume 1:1 ratio mixing,
Under conditions of ultrasonic power 150W, then ultrasonic wave added demulsification 0.5h centrifuges 10min in 5000r/min, obtains positioned at upper layer
Demulsification oil；The free oil II and the demulsification oil that free oil I that step 2) obtains, step 4) are obtained merge, as setose abelmoschus seed
Oil.
Among the present embodiment, free oil recovery rate is 85%, percent of total oil 92%；Protein recovery is 85%,
Setose abelmoschus seed lipidated protein is 87%.The rate of abolishing of ultrasonic wave added ethyl alcohol demulsification method emulsion may be up to 91~92%.
Embodiment 4
1) setose abelmoschus seed screened successively, cleaned, 50 DEG C of forced air dryings for 24 hours, using medicinal herb grinder high speed crush,
40 mesh sieve is crossed, the setose abelmoschus seed powder is obtained；Setose abelmoschus seed powder and water are pressed 1:The solid-liquid ratio of 4 (w/v) mixes, and adjusts pH
Value is 7.5, adds the complex enzyme for being equivalent to setose abelmoschus seed powder quality 1%, and the complex enzyme includes that mass ratio is 2:1:1
Cellulase, pectase and protease；Then under conditions of 30 DEG C of temperature, ultrasonic power 150W, ultrasonic wave added enzyme digestion reaction
240min obtains enzymolysis liquid；
2) enzymolysis liquid for obtaining step 1) 2500r/min centrifuge 15min, the free oil I being distributed from top to bottom,
Emulsion I, protein liquid and waste residue；
3) after taking the protein liquid suitably dilution that step 2) obtains, molecular cut off is used to be surpassed for the ultrafiltration membrane of 10kDa
Filter, takes trapped fluid to be concentrated under reduced pressure, and is freeze-dried 48h, as setose abelmoschus seed protein；
4) it takes the emulsion I that step 2) obtains to centrifuge 10min in 10000r/min, obtains II He of free oil positioned at upper layer
Positioned at the emulsion II of lower layer；
5) ethyl alcohol of the emulsion II for obtaining step 4) and volumetric concentration 20% by volume 1:1 ratio mixing,
Under conditions of ultrasonic power 150W, then ultrasonic wave added demulsification 0.5h centrifuges 10min in 5000r/min, obtains positioned at upper layer
Demulsification oil；The free oil II and the demulsification oil that free oil I that step 2) obtains, step 4) are obtained merge, as setose abelmoschus seed
Oil.
Among the present embodiment, free oil recovery rate is 84%, percent of total oil 93%；Protein recovery is 86%,
Setose abelmoschus seed lipidated protein is 87%.The rate of abolishing of ultrasonic wave added ethyl alcohol demulsification method emulsion may be up to 91~92%.
Embodiment 5
1) setose abelmoschus seed screened successively, cleaned, 50 DEG C of forced air dryings for 24 hours, using medicinal herb grinder high speed crush,
40 mesh sieve is crossed, the setose abelmoschus seed powder is obtained；Setose abelmoschus seed powder and water are pressed 1:The solid-liquid ratio of 4 (w/v) mixes, and adjusts pH
Value is 7.5, adds the complex enzyme for being equivalent to setose abelmoschus seed powder quality 1%, and the complex enzyme includes that mass ratio is 2:1:1
Cellulase, pectase and protease；Then under conditions of temperature 50 C, ultrasonic power 200W, ultrasonic wave added enzyme digestion reaction
180min obtains enzymolysis liquid；
2) enzymolysis liquid for obtaining step 1) 2500r/min centrifuge 15min, the free oil I being distributed from top to bottom,
Emulsion I, protein liquid and waste residue；
3) after taking the protein liquid suitably dilution that step 2) obtains, molecular cut off is used to be surpassed for the ultrafiltration membrane of 10kDa
Filter, takes trapped fluid to be concentrated under reduced pressure, and is freeze-dried 48h, as setose abelmoschus seed protein；
4) it takes the emulsion I that step 2) obtains to centrifuge 10min in 10000r/min, obtains II He of free oil positioned at upper layer
Positioned at the emulsion II of lower layer；
5) ethyl alcohol of the emulsion II for obtaining step 4) and volumetric concentration 20% by volume 1:1 ratio mixing,
Under conditions of ultrasonic power 150W, then ultrasonic wave added demulsification 0.5h centrifuges 10min in 5000r/min, obtains positioned at upper layer
Demulsification oil；The free oil II and the demulsification oil that free oil I that step 2) obtains, step 4) are obtained merge, as setose abelmoschus seed
Oil.
Among the present embodiment, free oil recovery rate is 87%, percent of total oil 92%；Protein recovery is 84%,
Setose abelmoschus seed lipidated protein is 89%.The rate of abolishing of ultrasonic wave added ethyl alcohol demulsification method emulsion may be up to 91~92%.
Embodiment 6
1) setose abelmoschus seed screened successively, cleaned, 75 DEG C of forced air dryings for 24 hours, using medicinal herb grinder high speed crush,
50 mesh sieve is crossed, the setose abelmoschus seed powder is obtained；Setose abelmoschus seed powder and water are pressed 1:The solid-liquid ratio of 6 (w/v) mixes, and adjusts pH
Value is 8.5, adds the complex enzyme for being equivalent to setose abelmoschus seed powder quality 3%, and the complex enzyme includes that mass ratio is 1:1:1:
1:1 cellulase, hemicellulase, pectase, zytase and protease；Then in 35 DEG C of temperature, ultrasonic power 100W
Under conditions of, ultrasonic wave added enzyme digestion reaction 100min obtains enzymolysis liquid；
2) enzymolysis liquid for obtaining step 1) 2000r/min centrifuge 20min, the free oil I being distributed from top to bottom,
Emulsion I, protein liquid and waste residue；
3) after taking the protein liquid suitably dilution that step 2) obtains, molecular cut off is used to be surpassed for the ultrafiltration membrane of 30kDa
Filter, takes trapped fluid to be concentrated under reduced pressure, and is freeze-dried 48h, as setose abelmoschus seed protein；
4) it takes the emulsion I that step 2) obtains to centrifuge 15min in 12000r/min, obtains II He of free oil positioned at upper layer
Positioned at the emulsion II of lower layer；
5) ethyl alcohol of the emulsion II for obtaining step 4) and volumetric concentration 40% by volume 1:2 ratio mixing,
Under conditions of ultrasonic power 100W, then ultrasonic wave added demulsification 1.5h centrifuges 15min in 6500r/min, obtains positioned at upper layer
Demulsification oil；The free oil II and the demulsification oil that free oil I that step 2) obtains, step 4) are obtained merge, as setose abelmoschus seed
Oil.
Among the present embodiment, free oil recovery rate is 83%, percent of total oil 95%；Protein recovery is 85%,
Setose abelmoschus seed lipidated protein is 90%.The rate of abolishing of ultrasonic wave added ethyl alcohol demulsification method emulsion may be up to 91~92%.
Embodiment 7
1) setose abelmoschus seed screened successively, cleaned, 65 DEG C of forced air dryings for 24 hours, using medicinal herb grinder high speed crush,
60 mesh sieve is crossed, the setose abelmoschus seed powder is obtained；Setose abelmoschus seed powder and water are pressed 1:The solid-liquid ratio of 8 (w/v) mixes, and adjusts pH
Value is 9.0, adds the complex enzyme for being equivalent to setose abelmoschus seed powder quality 5%, and the complex enzyme includes that mass ratio is 1:1:1:2
Cellulase, hemicellulase, pectase and protease；Then under conditions of temperature 45 C, ultrasonic power 150W, ultrasound
Assistance enzymolysis reacts 200min, obtains enzymolysis liquid；
2) enzymolysis liquid for obtaining step 1) 3000r/min centrifuge 10min, the free oil I being distributed from top to bottom,
Emulsion I, protein liquid and waste residue；
3) after taking the protein liquid suitably dilution that step 2) obtains, molecular cut off is used to be surpassed for the ultrafiltration membrane of 30kDa
Filter, takes trapped fluid to be concentrated under reduced pressure, and is freeze-dried 48h, as setose abelmoschus seed protein；
4) it takes the emulsion I that step 2) obtains to centrifuge 12min in 15000r/min, obtains II He of free oil positioned at upper layer
Positioned at the emulsion II of lower layer；
5) ethyl alcohol of the emulsion II for obtaining step 4) and volumetric concentration 60% by volume 1:3 ratio mixing,
Under conditions of ultrasonic power 120W, then ultrasonic wave added demulsification 1h centrifuges 12min in 8000r/min, obtains positioned at the broken of upper layer
Missible oil；The free oil II and the demulsification oil that free oil I that step 2) obtains, step 4) are obtained merge, as setose abelmoschus seed oil.
Among the present embodiment, free oil recovery rate is 85%, percent of total oil 90%；Protein recovery is 87%,
Setose abelmoschus seed lipidated protein is 88%.The rate of abolishing of ultrasonic wave added ethyl alcohol demulsification method emulsion may be up to 91~92%.
Embodiment 8
1) setose abelmoschus seed screened successively, cleaned, 65 DEG C of forced air dryings for 24 hours, using medicinal herb grinder high speed crush,
60 mesh sieve is crossed, the setose abelmoschus seed powder is obtained；Setose abelmoschus seed powder and water are pressed 1:The solid-liquid ratio of 8 (w/v) mixes, and adjusts pH
Value is 9.0, adds the complex enzyme for being equivalent to setose abelmoschus seed powder quality 5%, and the complex enzyme includes that mass ratio is 1:1:1:2
Cellulase, hemicellulase, pectase and protease；Then under conditions of temperature 45 C, ultrasonic power 150W, ultrasound
Assistance enzymolysis reacts 200min, obtains enzymolysis liquid；
2) enzymolysis liquid for obtaining step 1) 3000r/min centrifuge 10min, the free oil I being distributed from top to bottom,
Emulsion I, protein liquid and waste residue；
3) after taking the protein liquid suitably dilution that step 2) obtains, molecular cut off is used to be surpassed for the ultrafiltration membrane of 30kDa
Filter, takes trapped fluid to be concentrated under reduced pressure, and is freeze-dried 48h, as setose abelmoschus seed protein；
4) it takes the emulsion I that step 2) obtains to centrifuge 12min in 15000r/min, obtains II He of free oil positioned at upper layer
Positioned at the emulsion II of lower layer；
5) ethyl alcohol of the emulsion II for obtaining step 4) and volumetric concentration 60% by volume 1:3 ratio mixing,
It is stirred 5min under 100r/min, 12min is centrifuged in 8000r/min, obtains the demulsification oil positioned at upper layer；Step 2) is obtained
Free oil I, the obtained free oil II of step 4) and the demulsification oil merge, as setose abelmoschus seed oil.
Among the present embodiment, free oil recovery rate is 84%, percent of total oil 87%；Protein recovery is 80%,
Setose abelmoschus seed lipidated protein is 88%.The rate of abolishing of ethyl alcohol demulsification method emulsion is 65~68%.
Embodiment 9
1) setose abelmoschus seed screened successively, cleaned, 65 DEG C of forced air dryings for 24 hours, using medicinal herb grinder high speed crush,
60 mesh sieve is crossed, the setose abelmoschus seed powder is obtained；Setose abelmoschus seed powder and water are pressed 1:The solid-liquid ratio of 8 (w/v) mixes, and adjusts pH
Value is 9.0, adds the complex enzyme for being equivalent to setose abelmoschus seed powder quality 5%, and the complex enzyme includes that mass ratio is 1:1:1:2
Cellulase, hemicellulase, pectase and protease；Then under conditions of 38 DEG C of temperature, enzyme digestion reaction 720min is obtained
To enzymolysis liquid；
2) enzymolysis liquid for obtaining step 1) 3000r/min centrifuge 10min, the free oil I being distributed from top to bottom,
Emulsion I, protein liquid and waste residue；
3) after taking the protein liquid suitably dilution that step 2) obtains, molecular cut off is used to be surpassed for the ultrafiltration membrane of 30kDa
Filter, takes trapped fluid to be concentrated under reduced pressure, and is freeze-dried 48h, as setose abelmoschus seed protein；
4) it takes the emulsion I that step 2) obtains to centrifuge 12min in 15000r/min, obtains II He of free oil positioned at upper layer
Positioned at the emulsion II of lower layer；
5) ethyl alcohol of the emulsion II for obtaining step 4) and volumetric concentration 60% by volume 1:3 ratio mixing,
Under conditions of ultrasonic power 120W, then ultrasonic wave added demulsification 1h centrifuges 12min in 8000r/min, obtains positioned at the broken of upper layer
Missible oil；The free oil II and the demulsification oil that free oil I that step 2) obtains, step 4) are obtained merge, as setose abelmoschus seed oil.
Among the present embodiment, free oil recovery rate is 68%, percent of total oil 76%；Protein recovery is 71%,
Setose abelmoschus seed lipidated protein is 85%.The rate of abolishing of ultrasonic wave added ethyl alcohol demulsification method emulsion may be up to 85~87%.
The above, only present pre-ferred embodiments, therefore cannot limit the scope of implementation of the present invention according to this, i.e., according to
Equivalent changes and modifications made by the scope of the claims of the present invention and description all should still belong in the range of the present invention covers.
Claims (7)
1. a kind of method of ultrasonic wave added aqueous enzymatic extraction setose abelmoschus seed oil and protein, it is characterised in that：Including：
1) setose abelmoschus seed powder and water are pressed 1:4~8 solid-liquid ratio mixing, it is 7.0~9.0 to adjust pH value, adds and is equivalent to
The complex enzyme of setose abelmoschus seed powder quality 1~5%, the complex enzyme include that cellulase, hemicellulase, pectase, wood are poly-
At least one of carbohydrase further includes protease, and the content of protease is 20~45% in complex enzyme；Then temperature 30~
60 DEG C, under conditions of 100~200W of ultrasonic power, ultrasonic wave added 60~240min of enzyme digestion reaction obtains enzymolysis liquid；
2) the enzymolysis liquid low-speed centrifugal for obtaining step 1), the free oil I being distributed from top to bottom, emulsion I, protein liquid and
Waste residue；
3) the protein liquid ultrafiltration that step 2) obtains is taken, trapped fluid concentration, drying, as setose abelmoschus seed protein are taken；
4) I high speed centrifugation of emulsion that step 2) obtains is taken, the emulsion positioned at the free oil II on upper layer and positioned at lower layer is obtained
Ⅱ；
5) ethyl alcohol of the emulsion II for obtaining step 4) and volumetric concentration 20~60% by volume 1:1~3 ratio mixing,
Under conditions of 100~150W of ultrasonic power, ultrasonic wave added 0.5~1.5h of demulsification, then medium-speed centrifuge, obtains demulsification oil；It will
The free oil II and the demulsification oil that free oil I that step 2) obtains, step 4) obtain merge, as setose abelmoschus seed oil.
2. according to the method described in claim 1, it is characterized in that：In the step 1), by setose abelmoschus seed screening, clean, 50
~75 DEG C of 22~26h of forced air drying smash it through at a high speed 40~60 mesh sieve, obtain the setose abelmoschus seed powder.
3. according to the method described in claim 1, it is characterized in that：In the step 1), complex enzyme includes that mass ratio is 1~3:
0.5~1.5:0.5~1.5 cellulase, pectase and protease.
4. according to the method described in claim 1, it is characterized in that：In the step 2), low-speed centrifugal be 2000~
3000r/min centrifuges 10~20min.
5. according to the method described in claim 1, it is characterized in that：In the step 3), use molecular cut off for 10~
The ultrafiltration membrane of 30kDa carries out ultrafiltration, and trapped fluid is taken to be concentrated under reduced pressure, and is freeze-dried 45~50h, obtains the setose abelmoschus seed albumen
Matter.
6. according to the method described in claim 1, it is characterized in that：In the step 4), high speed centrifugation be 10000~
15000r/min centrifuges 10~15min.
7. according to the method described in claim 1, it is characterized in that：In the step 5), medium-speed centrifuge be 5000~
8000r/min centrifuges 10~15min.
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