CN107373011A - Method of protein in one kind extraction rapeseed dregs - Google Patents
Method of protein in one kind extraction rapeseed dregs Download PDFInfo
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- CN107373011A CN107373011A CN201710577738.8A CN201710577738A CN107373011A CN 107373011 A CN107373011 A CN 107373011A CN 201710577738 A CN201710577738 A CN 201710577738A CN 107373011 A CN107373011 A CN 107373011A
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- 235000004977 Brassica sinapistrum Nutrition 0.000 title claims abstract description 84
- 238000000605 extraction Methods 0.000 title claims abstract description 56
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 56
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 56
- 238000000034 method Methods 0.000 title claims abstract description 26
- 244000188595 Brassica sinapistrum Species 0.000 title 1
- 240000002791 Brassica napus Species 0.000 claims abstract description 83
- 241000894006 Bacteria Species 0.000 claims abstract description 32
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 32
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims abstract description 32
- 239000007788 liquid Substances 0.000 claims abstract description 25
- 238000000855 fermentation Methods 0.000 claims abstract description 24
- 230000004151 fermentation Effects 0.000 claims abstract description 24
- 239000000203 mixture Substances 0.000 claims abstract description 22
- 102000004190 Enzymes Human genes 0.000 claims abstract description 21
- 108090000790 Enzymes Proteins 0.000 claims abstract description 21
- 239000000843 powder Substances 0.000 claims abstract description 21
- 240000006439 Aspergillus oryzae Species 0.000 claims abstract description 18
- 235000002247 Aspergillus oryzae Nutrition 0.000 claims abstract description 18
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 18
- 241000235342 Saccharomycetes Species 0.000 claims abstract description 16
- 239000004310 lactic acid Substances 0.000 claims abstract description 16
- 235000014655 lactic acid Nutrition 0.000 claims abstract description 16
- 241000193830 Bacillus <bacterium> Species 0.000 claims abstract description 15
- 230000010355 oscillation Effects 0.000 claims abstract description 13
- 238000005406 washing Methods 0.000 claims abstract description 11
- 241000228212 Aspergillus Species 0.000 claims abstract description 9
- 235000019779 Rapeseed Meal Nutrition 0.000 claims abstract description 9
- 238000001914 filtration Methods 0.000 claims abstract description 9
- 239000004456 rapeseed meal Substances 0.000 claims abstract description 9
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims abstract description 8
- 239000012141 concentrate Substances 0.000 claims abstract description 7
- 229940088598 enzyme Drugs 0.000 claims description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 14
- 108091005804 Peptidases Proteins 0.000 claims description 13
- 239000004365 Protease Substances 0.000 claims description 13
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims description 13
- 108010059892 Cellulase Proteins 0.000 claims description 9
- 229940106157 cellulase Drugs 0.000 claims description 9
- 108010062085 ligninase Proteins 0.000 claims description 7
- 238000002156 mixing Methods 0.000 claims description 7
- 238000004140 cleaning Methods 0.000 claims description 6
- 238000001556 precipitation Methods 0.000 claims description 6
- 230000001954 sterilising effect Effects 0.000 claims description 3
- 238000010438 heat treatment Methods 0.000 claims description 2
- 244000025254 Cannabis sativa Species 0.000 claims 1
- 238000000354 decomposition reaction Methods 0.000 abstract description 4
- 239000008236 heating water Substances 0.000 abstract description 2
- 230000001376 precipitating effect Effects 0.000 abstract 1
- 235000018102 proteins Nutrition 0.000 description 45
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 17
- 239000002904 solvent Substances 0.000 description 8
- 239000003513 alkali Substances 0.000 description 5
- 235000016709 nutrition Nutrition 0.000 description 5
- 238000004090 dissolution Methods 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 239000003921 oil Substances 0.000 description 4
- 238000000751 protein extraction Methods 0.000 description 4
- 238000011084 recovery Methods 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 3
- 150000001413 amino acids Chemical class 0.000 description 3
- 238000004925 denaturation Methods 0.000 description 3
- 230000036425 denaturation Effects 0.000 description 3
- 239000004615 ingredient Substances 0.000 description 3
- 235000004252 protein component Nutrition 0.000 description 3
- 230000002797 proteolythic effect Effects 0.000 description 3
- 235000013311 vegetables Nutrition 0.000 description 3
- 244000063299 Bacillus subtilis Species 0.000 description 2
- 235000014469 Bacillus subtilis Nutrition 0.000 description 2
- 240000007594 Oryza sativa Species 0.000 description 2
- 235000007164 Oryza sativa Nutrition 0.000 description 2
- 210000000481 breast Anatomy 0.000 description 2
- 210000002421 cell wall Anatomy 0.000 description 2
- 229920002678 cellulose Polymers 0.000 description 2
- 239000001913 cellulose Substances 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 230000004927 fusion Effects 0.000 description 2
- 230000003834 intracellular effect Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000002068 microbial inoculum Substances 0.000 description 2
- 239000012046 mixed solvent Substances 0.000 description 2
- 230000035764 nutrition Effects 0.000 description 2
- 230000017854 proteolysis Effects 0.000 description 2
- 235000009566 rice Nutrition 0.000 description 2
- 150000003384 small molecules Chemical class 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 108010082495 Dietary Plant Proteins Proteins 0.000 description 1
- 238000003916 acid precipitation Methods 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000007797 corrosion Effects 0.000 description 1
- 238000005260 corrosion Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 239000008157 edible vegetable oil Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002481 ethanol extraction Methods 0.000 description 1
- 239000004519 grease Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 235000008935 nutritious Nutrition 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
- 210000004885 white matter Anatomy 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/14—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from leguminous or other vegetable seeds; from press-cake or oil-bearing seeds
- A23J1/148—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from leguminous or other vegetable seeds; from press-cake or oil-bearing seeds by treatment involving enzymes or microorganisms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Abstract
The invention discloses one kind to extract method of protein in rapeseed dregs, it is characterised in that including following aspect：（1）Crush, the rapeseed dregs after extracting will be squeezed through crushing；（2）Enzymolysis, mixed enzyme solution is added into rapeseed dregs powder, and constant temperature digests at a temperature of 42 50 DEG C；（3）Ferment, be separately added into lactic acid bacteria and saccharomycete mix bacterium agent, aspergillus oryzae and hay bacillus mix bacterium agent in rapeseed dregs enzymolysis liquid, carry out fermentation decomposition；（4）Extraction, the ethanol of its quality 40% is added into Rapeseed Meal by Aspergillus Fermentation liquid, through supersonic oscillations and heating water bath, centrifuge, extraction concentrate is made after low temperature concentration；（5）Washing, rapeseed dregs add ethanol repeated washing 2 times in precipitating；（6）Albumen powder is produced, the hydrochloric acid solution that mass concentration is 12% is added, adjusts pH value of solution 34, protein powder is made after centrifuging, filtering and dry.
Description
Technical field
The invention belongs to rapeseed Processing technical field, and in particular to method of protein in one kind extraction rapeseed dregs.
Background technology
Rapeseed is the main oil crops in China, and the cultivated area of rape accounts for more than the 40% of the oil crops gross area,
Yield is more than the 30% of oil plant total output, occupies first place in the world.Containing abundant nutritional ingredient in rapeseed, wherein containing containing grease
Measure 35%-45%, protein content 25%-32%；And contain abundant protein, vegetable seed in the rapeseed dregs after squeezing is extracted
Albumen is a kind of full price albumen, has preferable amino acid compositional model, several amino acids composition can be formed through enzymolysis, its
The abundant source of protein can be turned into, the protein extracted can be used as the nutrition adding ingredients such as food, feed, such as by rapeseed dregs
Being abandoned as discarded object can cause ample resources to waste.And rapeseed dregs is extracted using sodium hydroxide, extraction pH is higher, alkali
Property corrosion it is strong, protein denaturation in rapeseed dregs can be caused, cause Protein Extraction amount in rapeseed dregs to decline, protein also loses original
Nutritious activity；And the rapeseed dregs after sodium hydroxide solution soaks, it contains substantial amounts of alkali composition after extraction separation, if not
Continue working process and remove sodium hydroxide to cause rapeseed dregs not continue with into branch, cause the wasting of resources, and working process
Use cost can be improved.Water enzyme extraction method, using cellulase, pectase and protease to rapeseed dregs dissolution, by the time,
The condition elements such as temperature are had a great influence, and extraction efficiency can be caused low；And water is used to be dissolved water-soluble as proteolytic agent
Property protein, rather than water soluble protein can not carry out effective extraction and application, and extraction rate of protein is low in rapeseed dregs.
The content of the invention
The present invention is directed to the problem of existing：Rapeseed dregs composition, vegetable seed are China main edible oil material crop, and yield accounts for oil
Expect the 1/3 of total output, to produce substantial amounts of rapeseed dregs material after annual squeezing extraction, utilization rate is low to cause the wasting of resources；And in vegetable seed
Protein content 25%-32%, can be that food and feed manufacturing etc. provide nutrition adding ingredient containing abundant protein.Alkali carries
Method, dissolved and extracted using sodium hydroxide, alkalescence is high, corrosivity is big, can cause protein denaturation in rapeseed dregs, and recovery rate declines；
And contain substantial amounts of sodium hydroxide in rapeseed dregs after extracting, cause rapeseed dregs not to be continuing with, and be processed meeting
Use cost is caused to improve.Water enzyme extraction method, using cellulase, pectase and protease to rapeseed dregs dissolution, by when
Between, the condition element such as temperature have a great influence, extraction efficiency can be caused low, and provided protease species are few, decomposition has
Limit；And water is used to dissolve water soluble protein, rather than water soluble protein can not be carried out effectively as proteolytic agent
Extraction and application, extraction rate of protein is low in rapeseed dregs.To solve the above problems, the invention provides one kind to extract egg in rapeseed dregs
The method of white matter.
The present invention is achieved by the following technical solutions：
Method of protein in one kind extraction rapeseed dregs, comprises the following steps:
（1）Crush：The rapeseed dregs after extracting will be squeezed through crushing, obtain rapeseed dregs powder；
（2）Enzymolysis：The mixed enzyme solution that mass concentration is 5%-6%, cellulase, pectase and ligninase are added into rapeseed dregs powder
Bosom can be carried out brokenly to plant cell wall, promotes intracellular nutritional composition dissolution, and protein degradation can be small molecule by protease
Structure, promote it to be dissolved in solvent, first vibrated in ultrasonic wave, constant temperature digests 3-4h at a temperature of 42-50 DEG C, and mixing speed is
23-29 turns/min, and supersonic oscillations, heated at constant temperature and stirring can increase each component molecules vigor in solution, improve enzyme to dish
Seed dregs of rice enzymolysis, rapeseed dregs enzymolysis liquid is made；
（3）Fermentation：Its quality 2%-3% lactic acid bacteria and saccharomycete mix bacterium agent is added into rapeseed dregs enzymolysis liquid, can be to vegetable seed
The compositions such as dregs of rice sugar, cellulose are degraded, and improve Protein Extraction purity, 8-10h is cultivated under 26-28 DEG C of constant temperature, sterilized
Add its quality 3%-4% aspergillus oryzae and hay bacillus mix bacterium agent after processing into rapeseed dregs enzymolysis liquid again, aspergillus oryzae and withered
Caused enzyme class is more in careless bacillus fermentation, content is high, can improve the decomposition to protein in rapeseed dregs, high molecular weight protein
Matter is degraded to water-soluble and Organic-soluble protein component, improves recovery rate, 10-14h is cultivated under 26-28 DEG C of constant temperature, is made
Rapeseed Meal by Aspergillus Fermentation liquid；
（4）Extraction：The ethanol of its quality 40% is added into Rapeseed Meal by Aspergillus Fermentation liquid, forms water and ethanol mixing Extraction solvent, can be same
When extraction is water-soluble and Organic-soluble protein, the alternate oscillation 15-20min under frequency 25kHz and 33kHz ultrasonic wave, juxtaposition
1-2h is heated under 33-36 DEG C of water bath with thermostatic control, improves solvent and solute molecule liveness, promotes mutually fusion, improves protein
Dissolving and extraction, extract solution and rapeseed dregs precipitation are made after centrifugal filtration；Extract solution is concentrated into former extracting liq in low temperature
Long-pending 1/6, obtain extraction concentrate；
（5）Washing：The ethanol that 1-2 times of its quality is added in being precipitated to rapeseed dregs is washed, repeated washing 2 times, to rapeseed dregs
Residual protein is extracted in precipitation, and cleaning solution is made after centrifugal filtration；
（6）Produce albumen powder：The hydrochloric acid solution that mass concentration is 12%, adjustment are added after cleaning solution and extraction concentrate are mixed
PH value of solution 3-4, protein powder is made after centrifuging, filtering and dry.
Step（2）Described mixed enzyme solution, it includes pectase, ligninase, cellulase and protease, itself and rapeseed dregs
The quality proportioning of powder is 4-6:1.
Step（2）The supersonic oscillations, its frequency are 25kHz, duration of oscillation 8-12min.
Step（3）Described lactic acid bacteria and saccharomycete mix bacterium agent, wherein lactic acid bacteria：Saccharomycete quality proportioning is 1:1；Institute
The aspergillus oryzae and hay bacillus mix bacterium agent stated, wherein aspergillus oryzae：Hay bacillus quality proportioning is 2-3:3-4.
Step（3）Described sterilization treatment, its temperature are 62-65 DEG C, time 20-30min.
The present invention has advantages below compared with prior art：Digest, lignoenzyme, pectase and cellulase in mixed enzyme
Cell can be decomposed, promote into and analyze, protease promotes part breaks down proteins, forms water-soluble and Organic-soluble albumen
Matter, amino acid；And supersonic oscillations and heated at constant temperature, each component molecules liveness in solution can be improved, promotes enzyme to rapeseed dregs
The enzymolysis of powder.Fermentation process, lactic acid bacteria and saccharomycetes to make fermentation are first added, the compositions such as rapeseed dregs sugar, cellulose can be carried out
Degraded, Protein Extraction purity is improved, and protease, the ligninase in metabolite carry out preliminary exposition to protein；And
Using aspergillus oryzae and hay bacillus, in its fermentation process, caused enzyme class is more, content is high, can improve to protease in rapeseed dregs
Solution acts on, and improves extraction rate of protein.Extracting method, protein after enzymolysis and fermentation is entered using the mixed solvent of second alcohol and water
Row extraction, water-soluble and Organic-soluble protein in rapeseed dregs can be extracted simultaneously, increases extraction rate of protein；And two kinds of frequencies
Ultrasonic wave alternate oscillation and heating water bath mode, solvent and solute molecule vigor are improved, promote mutually oozing between solvent and solute
Effect thoroughly, improve Protein Extraction efficiency.Washing methods, rapeseed dregs precipitation is reused ethanol and washed after being filtered to extraction
Extraction, residual protein during alcohol soluble rapeseed dregs precipitates, increase extraction rate of protein.The inventive method by enzyme solution and
Fermentation process is combined, and solves the defects of protease species is few, enzymolysis is limited in enzyme solution, and avoids use
Alkali lye denaturation to caused by protein, improves extraction rate of protein and activity；Mixed solvent is used as using water and ethanol
Protein is extracted jointly, water-soluble and Organic-soluble protein component is can extract, increases and protein in rapeseed dregs is carried
Take rate.
Embodiment
Embodiment 1：
Method of protein in one kind extraction rapeseed dregs, comprises the following steps:
（1）Crush：The rapeseed dregs after extracting will be squeezed through crushing, obtain rapeseed dregs powder；
（2）Enzymolysis：The mixed enzyme solution that mass concentration is 5.2%, cellulase, pectase and ligninase are added into rapeseed dregs powder
Bosom can be carried out brokenly to plant cell wall, promotes intracellular nutritional composition dissolution, and protein degradation can be small molecule by protease
Structure, promote it to be dissolved in solvent, first vibrated in ultrasonic wave, constant temperature digests 3.5h, mixing speed 25 at a temperature of 44 DEG C
Turn/min, supersonic oscillations, heated at constant temperature and stirring can increase each component molecules vigor in solution, improve enzyme to rapeseed dregs enzyme
Solution acts on, and rapeseed dregs enzymolysis liquid is made；
（3）Fermentation：Lactic acid bacteria and the saccharomycete mix bacterium agent of its quality 2.4% are added into rapeseed dregs enzymolysis liquid, in 27 DEG C of constant temperature
Lower culture 8.5h, add aspergillus oryzae and the hay bacillus mixing of its quality 3.5% after sterilized processing into rapeseed dregs enzymolysis liquid again
Caused enzyme class is more in microbial inoculum, aspergillus oryzae and fermenting bacillus subtilis, content is high, can improve the decomposition to protein in rapeseed dregs
Effect, macro-molecular protein are degraded to water-soluble and Organic-soluble protein component, improve recovery rate, are cultivated under 27 DEG C of constant temperature
11h, Rapeseed Meal by Aspergillus Fermentation liquid is made；
（4）Extraction：The ethanol of its quality 40% is added into Rapeseed Meal by Aspergillus Fermentation liquid, forms water and ethanol mixing Extraction solvent, can be same
When extraction is water-soluble and Organic-soluble protein, the alternate oscillation 16min under frequency 25kHz and 33kHz ultrasonic wave, be placed in 34
1.5h is heated under DEG C water bath with thermostatic control, improves solvent and solute molecule liveness, promotes mutually fusion, proteolytic is improved and carries
Take, extract solution and rapeseed dregs precipitation are made after centrifugal filtration；Extract solution is concentrated into the 1/6 of former extracting liquid volume in low temperature,
Concentrate must be extracted；
（5）Washing：The ethanol that 1-2 times of its quality is added in being precipitated to rapeseed dregs is washed, repeated washing 2 times, to rapeseed dregs
Residual protein is extracted in precipitation, and cleaning solution is made after centrifugal filtration；
（6）Produce albumen powder：The hydrochloric acid solution that mass concentration is 12%, adjustment are added after cleaning solution and extraction concentrate are mixed
PH value of solution 3-4, protein powder is made after centrifuging, filtering and dry.
Step（2）Described mixed enzyme solution, it includes pectase, ligninase, cellulase and protease, itself and rapeseed dregs
The quality proportioning of powder is 4:1.
Step（2）The supersonic oscillations, its frequency are 25kHz, duration of oscillation 9min.
Step（3）Described lactic acid bacteria and saccharomycete mix bacterium agent, wherein lactic acid bacteria：Saccharomycete quality proportioning is 1:1；Institute
The aspergillus oryzae and hay bacillus mix bacterium agent stated, wherein aspergillus oryzae：Hay bacillus quality proportioning is 2:3.
Step（3）Described sterilization treatment, its temperature are 63 DEG C, time 24min.
Embodiment 2：
Compared with embodiment 1, step changes in the following areas the present embodiment 2：
Step（2）Enzymolysis, its method are：
The mixed enzyme solution that mass concentration is 5.8% is added into rapeseed dregs powder, first 11min is vibrated in 25kHz ultrasonic waves, 47
Constant temperature digests 4h at a temperature of DEG C, and mixing speed is 27 turns/min, and rapeseed dregs enzymolysis liquid is made.
Step（2）Described mixed enzyme solution, it includes pectase, ligninase, cellulase and protease, itself and rapeseed dregs
The quality proportioning of powder is 6:1.
Step（3）Fermentation, its method are：
Lactic acid bacteria and the saccharomycete mix bacterium agent of its quality 2.7% are added into rapeseed dregs enzymolysis liquid, is cultivated under 28 DEG C of constant temperature
9h, sterilize 28min at a temperature of 64 DEG C, then adds the aspergillus oryzae of its quality 3.6% into rapeseed dregs enzymolysis liquid and hay bacillus mixes
Microbial inoculum is closed, 13h is cultivated under 27 DEG C of constant temperature, Rapeseed Meal by Aspergillus Fermentation liquid is made.
Step（3）Described lactic acid bacteria and saccharomycete mix bacterium agent, wherein lactic acid bacteria：Saccharomycete quality proportioning is 1:1；Institute
The aspergillus oryzae and hay bacillus mix bacterium agent stated, wherein aspergillus oryzae：Hay bacillus quality proportioning is 3:4.
Step（4）Extraction, its method are：
The ethanol of its quality 40% is added into Rapeseed Meal by Aspergillus Fermentation liquid, the alternate oscillation under frequency 25kHz and 33kHz ultrasonic wave
19min, it is placed in heating 2h under 35 DEG C of waters bath with thermostatic control.
Contrast 1：
This contrast 1 does not carry out step compared with embodiment 1（2）Enzymolysis, other steps are same as Example 1.
Contrast 2：
This contrast 2 does not carry out step compared with embodiment 1（3）Lactic acid bacteria and saccharomycete add in fermentation, and other steps are with implementing
Example 1 is identical.
Contrast 3：
This contrast 3 does not carry out step compared with embodiment 1（3）Aspergillus oryzae and hay bacillus add in fermentation, other steps and reality
It is identical to apply example 1.
Contrast 4：
This contrast 4 does not carry out step compared with embodiment 2（4）Use ethanol in extraction, other steps are same as Example 2.
Contrast 5：
This contrast 5 does not carry out step compared with embodiment 2（5）Washing, other steps are same as Example 2.
Control group：
Fermentation and ethanol extraction is not used using alkali carries acid-precipitation method extraction protein in control group.
Experimental data：
| Project | Extraction rate of protein % | DNA purity % | Rapeseed dregs residue pH |
| Embodiment 1 | 72.35% | 94.53% | 6.2 |
| Embodiment 2 | 72.13% | 94.21% | 6.3 |
| Contrast 1 | 68.61% | 94.19% | 6.8 |
| Contrast 2 | 69.84% | 91.21% | 6.5 |
| Contrast 3 | 65.21% | 92.38% | 6.9 |
| Contrast 4 | 69.30% | 89.64% | 7.1 |
| Contrast 5 | 71.38% | 93.47% | 7.3 |
| Control group | 54.76% | 82.75% | 9.7 |
Synthesis result：The present invention is to protein extracting method in rapeseed dregs, and it is 17.59% that can improve extraction rate of protein, and extraction is pure
Degree improves 11.78%, reduces the use of acid-base reagent, and rapeseed dregs can be used directly after extraction, reduces extraction processing cost
16.4%.Using lactic acid bacteria and saccharomycetes to make fermentation, extraction rate of protein 2.51% can be improved, DNA purity improves 3.32%；And use
Aspergillus oryzae and fermenting bacillus subtilis method, can improve extraction rate of protein 7.14%, and DNA purity improves 2.15%；Added in extraction
It is 2.83% that ethanol, which can improve recovery rate, and DNA purity improves 4.57%.
Claims (7)
1. method of protein in one kind extraction rapeseed dregs, it is characterised in that comprise the following steps:
（1）Crush：The rapeseed dregs after extracting will be squeezed through crushing, obtain rapeseed dregs powder；
（2）Enzymolysis：The mixed enzyme solution that mass concentration is 5%-6% is added into rapeseed dregs powder, is first vibrated in ultrasonic wave, in 42-
Constant temperature digests 3-4h at a temperature of 50 DEG C, and mixing speed is that 23-29 turns/min, and rapeseed dregs enzymolysis liquid is made；
（3）Fermentation：Its quality 2%-3% lactic acid bacteria and saccharomycete mix bacterium agent is added into rapeseed dregs enzymolysis liquid, at 26-28 DEG C
8-10h is cultivated under constant temperature, adds its quality 3%-4% aspergillus oryzae and withered grass bar after sterilized processing into rapeseed dregs enzymolysis liquid again
Bacterium mix bacterium agent, 10-14h is cultivated under 26-28 DEG C of constant temperature, Rapeseed Meal by Aspergillus Fermentation liquid is made；
（4）Extraction：The ethanol of its quality 40% is added into Rapeseed Meal by Aspergillus Fermentation liquid, is handed under frequency 25kHz and 33kHz ultrasonic wave
For vibration 15-20min, it is placed in heating 1-2h under 33-36 DEG C of water bath with thermostatic control, extract solution and rapeseed dregs is made after centrifugal filtration
Precipitation；Extract solution low temperature is concentrated into the 1/6 of former extracting liquid volume, obtains extraction concentrate；
（5）Washing：The ethanol that 1-2 times of its quality is added in being precipitated to rapeseed dregs is washed, repeated washing 2 times, through centrifuging
Cleaning solution is made after filter；
（6）Produce albumen powder：The hydrochloric acid solution that mass concentration is 12%, adjustment are added after cleaning solution and extraction concentrate are mixed
PH value of solution 3-4, protein powder is made after centrifuging, filtering and dry.
2. method of protein in extraction rapeseed dregs as claimed in claim 1, it is characterised in that step（2）Described mixed enzyme
Liquid, it includes pectase, ligninase, cellulase and protease, and the quality proportioning of itself and rapeseed dregs powder is 4-6:1.
3. method of protein in extraction rapeseed dregs as claimed in claim 1, it is characterised in that step（2）The ultrasonic wave shakes
Swing, its frequency is 25kHz, duration of oscillation 8-12min.
4. method of protein in extraction rapeseed dregs as claimed in claim 1, it is characterised in that step（3）Described lactic acid bacteria
With saccharomycete mix bacterium agent, wherein lactic acid bacteria：Saccharomycete quality proportioning is 1:1；Described aspergillus oryzae and hay bacillus Mixed Microbes
Agent, wherein aspergillus oryzae：Hay bacillus quality proportioning is 2-3:3-4.
5. method of protein in extraction rapeseed dregs as claimed in claim 1, it is characterised in that step（3）At described sterilizing
Reason, its temperature are 62-65 DEG C, time 20-30min.
6. method of protein in extraction rapeseed dregs as claimed in claim 1, it is characterised in that step（4）The low temperature concentration,
Its temperature is 37-42 DEG C, time 40-50min.
7. method of protein in extraction rapeseed dregs as claimed in claim 1, it is characterised in that step（5）The washing, it is washed
Wash 33-36 DEG C of temperature, wash time 0.5-1h.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201710577738.8A CN107373011A (en) | 2017-07-15 | 2017-07-15 | Method of protein in one kind extraction rapeseed dregs |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710577738.8A CN107373011A (en) | 2017-07-15 | 2017-07-15 | Method of protein in one kind extraction rapeseed dregs |
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| Publication Number | Publication Date |
|---|---|
| CN107373011A true CN107373011A (en) | 2017-11-24 |
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| CN201710577738.8A Pending CN107373011A (en) | 2017-07-15 | 2017-07-15 | Method of protein in one kind extraction rapeseed dregs |
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| CN107522767A (en) * | 2017-10-13 | 2017-12-29 | 陈增光 | A kind of preparation method of turmeric saponin |
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