CN105918606A - Extracting method of proteins in green tea dregs - Google Patents
Extracting method of proteins in green tea dregs Download PDFInfo
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- CN105918606A CN105918606A CN201610313547.6A CN201610313547A CN105918606A CN 105918606 A CN105918606 A CN 105918606A CN 201610313547 A CN201610313547 A CN 201610313547A CN 105918606 A CN105918606 A CN 105918606A
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Classifications
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- 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
- A23J1/001—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from waste materials, e.g. kitchen waste
- A23J1/005—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from waste materials, e.g. kitchen waste from vegetable waste materials
-
- 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
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Biochemistry (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Peptides Or Proteins (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The present invention discloses an extracting method of proteins in green tea dregs. The extracting method comprises the following steps: S1. enzymolysis: the green tea dregs are subjected to enzymolysis and wall breaking treatment by using the enzymolysis technology; S2. alkaline extraction: the enzymolyzed green tea dregs are continuously subjected to the alkaline extraction to extract green tea proteins to obtain an green tea protein extract; S3. acid precipitation: the obtained green tea protein extract in the S2 is subjected to the acid precipitation to obtain a precipitate; S4. desalting: the obtained precipitate in the S3 is subjected to desalting; and S5. decolorization: 75% acetoneis used to conduct the decolorization of the desalted green tea proteins in the S4 to obtain green tea proteins. The enzyme and alkali compound extraction method combines the enzymatic method and the alkali method. Firstly, the enzymes are used to conduct wall breaking treatment of the green tea dreg cells, which is beneficial for the protein outflow, and then the alkali method is used to conduct extraction of the tea proteins. Under the technology, more green tea proteins can be extracted only under low concentration of alkali, which is conductive for improving the extraction rate of the green tea proteins and preventing the loss of nutrients in the green tea proteins.
Description
Technical field
The invention belongs to tea protein extracting process field.More particularly, to the extracting method of albumen in a kind of green tea tea grounds.
Background technology
China is Tea Production state maximum in the world, along with the continuous expansion of deep processing field of tea, and the volume of production rapid development of the product such as tea beverage, instant tea and tea polyphenols.Tea beverage name as state's drink is classified as one of big beverage in the world three.After the nineties in 20th century, along with membrane technology, Freeze Drying Technique etc. are applied to production of instant tea, its production also reaches its maturity, and market gradually increases.Prominent physiological function based on tea polyphenols, its yield the most progressively increases, existing certain market scale.Either the extraction of the functional component such as tea beverage, instant tea or tea polyphenols all can produce substantial amounts of tea grounds.
Containing more available composition in Folium Camelliae sinensis, but the effective ingredient in the tea products such as tea beverage the most only accounts for a part for Folium Camelliae sinensis.The material that the deep processing such as tea beverage, instant tea is extracted comprehensively the most only accounts for about the 30% of dry weight of tea leaves, and the composition utilized is mainly tea polyphenols, caffeine, saccharide, amino acid and vitamin etc..In Folium Camelliae sinensis, protein content is about 20 ~ 30%, mainly includes glutelin, alcohol soluble protein, albumin and globulin, and the most only 1 ~ 2% can be dissolved in water, and the overwhelming majority can not be dissolved in water.Still more nutrient protein composition is remained in discarded tea grounds.
China's recycling to tea grounds at present is still in the preliminary study stage, and existing research is concentrated mainly on the component analysis of tea grounds, preliminary the extracting of tea albumen, tea grounds are used as the aspect such as research of feedstuff, fertilizer.And wherein, the research for the extraction process of tea albumen is the most less.In Folium Camelliae sinensis, water-soluble protein content is little, and the research to it is the most little.And about the research of Water insoluble tea protein, it is limited to extraction and the modification in test chamber stage at present, at present, modal Water insoluble tea protein extracting method is alkaline process.Although traditional alkaline process has the advantage that extraction ratio is high, but during extracting, concentration of lye is unsuitable too high, otherwise can cause the loss of green tea albumen nutrient substance；But on the other hand, it is desirable to utilize alkalinity extraction tea albumen, must carry out again obtaining higher albumen yield in the environment of alkali concn is higher；Therefore there is contradiction, process conditions are difficult to control, and extraction efficiency is difficult to stability contorting.And, still not for the specific aim Study on extraction of different types of Folium Camelliae sinensis, extraction and application inefficiency in prior art.
Summary of the invention
The technical problem to be solved in the present invention is defect and the deficiency overcoming existing tea grounds protein extraction to utilize technique, it is provided that the extracting method of albumen in the extracting method of a kind of tea albumen, especially green tea tea grounds, the natural tea leaf protein extracted from discarded green tea tea grounds.The method is enzyme alkali compound collecting method, with discarded green tea tea grounds as raw material, carry through enzymolysis, alkali, the step such as acid is sunk, desalination, decolouring, green tea egg white matter is extracted, this compound collecting method can be greatly improved the extraction ratio of green tea albumen, and desirably prevents the loss of green tea albumen nutrient substance.
Above-mentioned purpose of the present invention is achieved through the following technical solutions:
In a kind of green tea tea grounds, the extracting method of albumen, comprises the steps:
S1. enzymolysis: use zymolysis technique green tea tea grounds to be carried out enzymolysis broken wall treatment, to improve the extraction ratio of green tea albumen；
S2. alkali carries: the green tea tea grounds after enzymolysis continues to use alkalinity extraction green tea albumen, obtains green tea protein extract；
S3. acid is heavy: S2 gained green tea protein extract carries out acid heavy, obtains precipitate；
S4. desalination: S3 gained precipitate is carried out desalination；
S5. decolouring: take the green tea albumen after S4 desalination, utilize 75% acetone to decolour, obtain green tea albumen.
Wherein it is preferred to, the enzyme that enzymolysis described in step S1 is used is composite plant hydrolytic enzyme.
Preferably, composite plant hydrolytic enzyme described in step S1 is that Novi believes composite plant hydrolytic enzyme Viscozyme L.
Preferably, step S2 alkali carries used alkali is sodium hydroxide solution.
It is highly preferred that the parameter of enzymolysis described in step S1 is: enzyme concentration is 2.0～3.0%, solid-to-liquid ratio (w/v) is 1:15～1:30, and enzymolysis pH is 3.0～4.0, and hydrolysis temperature is 40～50 DEG C, and enzymolysis time is 1～3h.
In the parameter of enzymolysis described in step S1, it is highly preferred that enzyme concentration is 3%.It is highly preferred that described solid-to-liquid ratio (w/v) is 1:20.It is highly preferred that described enzymolysis pH is 3.4.It is highly preferred that described hydrolysis temperature is 45 DEG C.It is highly preferred that described enzymolysis time is 3h.
Furthermore it is preferred that the parameter that alkali described in step S2 carries is: concentration of lye is 0.08～0.12mol/L, solid-to-liquid ratio (w/v) is 1:40～1:60, and Extracting temperature is 70～90 DEG C, and extraction time is 60～120min；It is centrifuged and obtains green tea protein extract.
In the parameter that alkali described in step S2 carries, it is highly preferred that described concentration of lye is 0.1mol/L.It is highly preferred that described solid-to-liquid ratio (w/v) is 1:50.It is highly preferred that described Extracting temperature is 90 DEG C.It is highly preferred that described extraction time is 100min.It is highly preferred that described being centrifuged is 3000～4000r/min to be centrifuged 10～20min.More preferably 3500r/min is centrifuged 15min.
Furthermore it is preferred that described in step S3, acid is heavy is to take green tea protein extract, it is 2～3 with dilute hydrochloric acid regulation pH, centrifugal after standing 20～40min, it is precipitated thing.
It is highly preferred that the pH with dilute hydrochloric acid regulation green tea protein extract is 3.It is highly preferred that described time of repose is 30min.It is highly preferred that described centrifugal be 3000～4000r/min to be centrifuged 10～20min.More preferably 3500r/min is centrifuged 15min.
Additionally, preferably, desalination described in step S4 is to add suitable quantity of water mixing in precipitate, inject in bag filter, it is placed in pure water dialysis more than 24h, period changes pure water 3～5 times, until the outer water colorless of bag filter and electrical conductivity are close to pure water, then 3000～4000r/min are centrifuged 10～20min and obtain the green tea albumen after desalination.
It is highly preferred that described suitable quantity of water refers to that precipitate is 1:1～1:2 with the volume ratio of water.
Dialysis 24～48h it is placed in pure water described in it is highly preferred that.
It is highly preferred that described being centrifuged is that 3500r/min is centrifuged 15min.
It addition, it is highly preferred that green tea tea grounds described in step S1 is the discarded green tea tea grounds after extracting.
It is further preferred that described discarded green tea tea grounds crosses 60 mesh sieves after also needing to pulverize.
Preferably, the amount ratio of green tea albumen after desalination described in step S5 and 75% acetone is: solid-to-liquid ratio 1:4～1:8.It is highly preferred that the amount ratio of green tea albumen after desalination described in step S5 and 75% acetone is: solid-to-liquid ratio 1:6.
Preferably, described in step S5, the method for decolouring is: take the green tea albumen after S4 desalination, add 75% acetone, concussion 10～20min(preferred 15min), centrifugal abandon supernatant, be repeated several times (preferably twice), with pure water and be centrifuged, be repeated several times (preferably twice) with clean acetone, obtain green tea albumen.
It addition, the application of said method also should be within protection scope of the present invention.
The method have the advantages that
The enzyme alkali compound collecting method of the present invention is coupling enzyme process and alkaline process, the most first extracts tea albumen with enzyme liquid, more again extracts tea albumen with alkali liquor.Composite plant hydrolytic enzyme is used first green tea tea grounds cell to be carried out broken wall treatment, be conducive to the outflow of protein, then use traditional alkaline process that tea albumen is extracted, under this technique, only need to can extract more green tea albumen under the conditions of the alkali of low concentration, be conducive to improving the extraction ratio of green tea albumen, and desirably prevent to reduce the loss of green tea albumen nutritional labeling.
As a example by emerald green tea tea grounds, using traditional alkaline process to extract, the extraction ratio of green tea albumen is 52.35%, uses enzyme alkali to be combined new technology and extracts, and the extraction ratio of green tea albumen can reach 74.27%, and extraction ratio adds 21.92%.
Accompanying drawing explanation
Figure 1For must be with nonessential aminoacid proportion in tea grounds albumenFigure。
Figure 2For three kinds of primary amino acid proportions in tea grounds albumenFigure。
Figure 3The technological process of green tea albumen is extracted for enzyme alkali composite algorithm described in embodiment 1Figure。
Figure 4For the enzyme concentration impact on extraction ratio.
Figure 5For the enzymolysis time impact on extraction ratio.
Figure 6For the concentration of lye impact on extraction ratio.
Figure 7The impact on extraction ratio of the middle solid-to-liquid ratio is proposed for alkali.
Figure 8For the impact on extraction ratio of the alkali temperature raising degree.
Figure 9The time impact on extraction ratio is proposed for alkali.
Detailed description of the invention
Below in conjunction with descriptionAccompanying drawingFurther illustrate the present invention with specific embodiment, but the present invention is not limited in any form by embodiment.Unless stated otherwise, the present invention uses reagent, method and apparatus are the art conventional reagent, method and apparatus.
Unless stated otherwise, following example agents useful for same and material are commercial.
Embodiment
1
The protein composition analysis of tea grounds albumen
1, by analysis,Such as table 1Shown in, tea leaf protein amino acid composition is analyzed, and in tea leaf protein, amino acid classes has 17 kinds, and aminoacid composition is complete, rich content, seven kinds of essential amino acids containing needed by human body, byFigure 1Shown in, it is necessary to aminoacid percentage is 42%, and content is of a relatively high.Its Glutamic Acid, leucine, aspartate content are the highest, are 63.6mg/g, 53.3mg/g, 52.5 mg/g. respectivelyAs Figure 2Shown in, it is 12.15%, 10.18%, 10.03% that these three aminoacid accounts for the ratio of total amino acids respectively, and it is 67.64% that other total amino acid contents account for the ratio of total amino acids.
Histidine is the essential amino acids during children's upgrowth and development, and arginine is premature infant institute essential amino acids, in tea grounds albumen, both accounts for 14.4 mg/g, 10.3 mg/g respectively, illustrates that the tea grounds albumen that this experiment is extracted has certain advantage for producing infant food；Lysine content in cereal foods is the lowest, and be easily destroyed in the course of processing and cause human body to lack lysine, but content is the abundantest in tea grounds albumen, up to 40.6mg/g, illustrate tea grounds albumen for the lysine in strengthening and supplementary cereal foods significant and value.
Essential amino acids is 0.73 with the ratio of non-essential amino acid content, higher than the 0.6 of FAO/WHO standard regulation；Essential amino acids content accounts for the 42% of total amino acid content, higher than the 40% of FAO/WHO standard regulation.Illustrate that tea grounds albumen is a kind of quality plant albumen source.
Table 1 tea leaf protein amino acid composition is analyzedTable 1
| Essential amino acids (EAA) | Content (mg/g) | Non essential amino acid (NEAA) | Content (mg/g) |
| Threonine (Thr) | 19.8 | Aspartic acid (Asp) | 52.5 |
| Valine (Val) | 37.5 | Glutamic acid (Glu) | 63.6 |
| Methionine (Met) | 11.8 | Serine (Ser) | 25.1 |
| Phenylalanine (Phe) | 29.1 | Glycine (Gly) | 35.7 |
| Isoleucine (Ile) | 28.6 | Arginine (Arg) | 10.3 |
| Leucine (Leu) | 53.3 | Alanine (Ala) | 40.7 |
| Lysine (Lys) | 40.6 | Tyrosine (Tyr) | 25.5 |
| Proline (Pro) | 32.2 | ||
| Hydroxyproline (Hpro) | 2.9 | ||
| Histidine (His) | 14.4 | ||
| TEAA | 220.7 | ||
| Total amino acids (TEAA) | 523.6 | ||
| EAA/NEAA | 0.73 | ||
| EAA/TAA | 0.42 |
FromTableIt can be seen that the first limiting amino acid that methionine is tea grounds albumen in 2, content is relatively fewer.Tea leaf protein CV is 0.28, SRC is 72.22, size according to SRC value, with the Chicken Albumin SRC of regulation under FAO/WHO mode standard be 84.09, soybean protein SRC be 61.22 to contrast, the scoring of this some protein is ranked up: Chicken Albumin ＞ tea leaf protein ＞ soybean protein, illustrates that extracted tea leaf protein after purification is a kind of protein comparing high-quality, have better nutritivity to be worth, its nutritive value is slightly below Chicken Albumin, higher than soybean protein.
Table 2The amino acid ratio coefficient of tea leaf protein
The extracting method of albumen in embodiment 2 green tea tea grounds
As Figure 3Shown in, the extracting method of albumen in green tea tea grounds, step is as follows:
(1) selecting discarded green tea tea grounds is raw material, crosses 60 mesh sieves, obtain tea grounds powder after pulverizing, being that 1:20 addition composite plant hydrolytic enzyme carries out enzymolysis breaking cellular wall by solid-to-liquid ratio (w/v), control enzyme concentration is 2.5%, and enzymolysis pH value is 3.4, hydrolysis temperature is at 45 DEG C, and enzymolysis time is 3h；
(2) using alkaline extraction to extract green tea albumen, concentration of lye is 0.1mol/L, and solid-to-liquid ratio (w/v) is 1:50, and Extracting temperature is 90 DEG C, and extraction time is that 100min, 3500r/min are centrifuged 15min and obtain green tea protein extract；
(3) regulating green tea protein extract pH with dilute hydrochloric acid is 2.5, and after standing 30min, 3500r/min is centrifuged 15min and is precipitated thing (being crude protein)；
(4) in precipitate, suitable quantity of water mixing is added, inject in bag filter, it is placed in pure water dialysis more than 24h, period changes pure water 3～5 times, until the outer water colorless of bag filter and electrical conductivity are close to pure water, then 3500r/min is centrifuged the green tea albumen after 15min obtains desalination (i.e. desalination albumen)；
(5) take the acetone that the green tea albumen after desalination adds 75%, shake about 15min, centrifugal abandon supernatant, be repeated twice, with pure water and be centrifuged, be repeated twice clean acetone, it is thus achieved that green tea albumen.
Embodiment
3
The extracting method of albumen in green tea tea grounds
The extracting method of albumen in green tea tea grounds, step is as follows:
(1) choosing through repeating extraction and again drying gold tawny daylily green tea tea grounds is raw material, crossing 60 mesh sieves after pulverizing by solid-to-liquid ratio (w/v) is that 1:30 addition composite plant hydrolytic enzyme carries out enzymolysis breaking cellular wall, and control enzyme concentration is 3%, and enzymolysis pH value is 4.0, hydrolysis temperature is at 50 DEG C, and enzymolysis time is 1h；After enzymolysis 1h, 3500r/min is centrifuged that 15min is centrifugal removes supernatant；
(2) by solid-to-liquid ratio (w/v) 1:60, adding concentration in the product of step (1) is the alkali liquor of 0.12mol/L, and at temperature is 70 DEG C, extraction 60min, 3500r/min are centrifuged 15min and obtain gold tawny daylily green tea protein extract；
(3) regulating gold tawny daylily green tea protein extract pH with dilute hydrochloric acid is 2.0, and after standing 30min, 3500r/min is centrifuged 15min and is precipitated thing；
(4) in precipitate, suitable quantity of water mixing is added, injecting in bag filter, be placed in pure water dialysis more than 24h, period changes pure water 3 times, until the outer water colorless of bag filter and electrical conductivity are close to pure water, then 3500r/min is centrifuged the golden tawny daylily green tea albumen after 15min obtains desalination；
(5) take the acetone that the golden tawny daylily green tea albumen after desalination adds 75%, shake about 15min, centrifugal abandon supernatant, be repeated twice, with pure water and be centrifuged, be repeated twice clean acetone, it is thus achieved that gold tawny daylily green tea albumen.
Embodiment
4
Enzyme alkali compound collecting method and the contrast of alkaline process
1, enzyme alkali compound collecting method
(1) choosing through repeating extraction and again drying emerald green tea tea grounds is raw material, crosses 60 mesh sieves after pulverizing, is that 1:20 addition composite plant hydrolytic enzyme carries out enzymolysis breaking cellular wall by solid-to-liquid ratio (w/v), control enzyme concentration 2.5%, enzymolysis pH value is 3.4, and hydrolysis temperature is at 45 DEG C, and enzymolysis time is 3h；After enzymolysis 3h, 3500r/min is centrifuged that 15min is centrifugal removes supernatant；
(2) in step (1) products therefrom, by solid-to-liquid ratio (w/v) 1:50, adding concentration is the alkali liquor of 0.1mol/L, and at temperature is 90 DEG C, extraction 100min, 3500r/min are centrifuged 15min and obtain emerald green tea protein extract；
(3) regulating emerald green tea protein extract pH with dilute hydrochloric acid is 2.5, and after standing 30min, 3500r/min is centrifuged 15min and is precipitated thing；
(4) in precipitate, suitable quantity of water mixing is added, injecting in bag filter, be placed in pure water dialysis more than 24h, period changes pure water 5 times, until the outer water colorless of bag filter and electrical conductivity are close to pure water, then 3500r/min is centrifuged the emerald green tea albumen after 15min obtains desalination；
(5) take the acetone that the emerald green tea albumen after desalination adds 75%, shake about 15min, centrifugal abandon supernatant, be repeated twice, with pure water and be centrifuged, be repeated twice clean acetone, it is thus achieved that emerald green tea albumen.
2, same with emerald green tea tea grounds as raw material, use independent alkalinity extraction tea albumen.
Concrete grammar, with above-mentioned enzyme alkali compound collecting method, only difference is that the enzymolysis step eliminated in step (1), and i.e. choosing through repeating extraction and again drying emerald green tea tea grounds is raw material, after crossing 60 mesh sieves, directly utilizes alkaline process and extracts after pulverizing.
3, result shows: with emerald green tea tea grounds as raw material, uses independent alkaline process to extract, and the extraction ratio of green tea albumen is 52.35%, uses enzyme alkali to be combined new technology and extracts, and the extraction ratio of green tea albumen can reach 74.27%, and extraction ratio adds 21.92%.
Embodiment
5
The cooperation that in green tea protein extracting method, enzymolysis and alkali carry
1, explored in the enzyme alkali compound collecting method of the present invention by contrast test, the mating reaction that enzymolysis and alkali carry, test is grouped as follows:
(1) group 1: operational approach is with embodiment 1；
(2) group 2: operational approach with embodiment 1, only difference is that order exchange step (1) and the enzymolysis of (2) and alkali proposed；I.e. selecting discarded green tea tea grounds is raw material, crosses 60 mesh sieves after pulverizing, after obtaining tea grounds powder, first with alkalinity extraction, is further continued for utilizing enzymatic isolation method to continue to extract.
2, result shows, the extraction ratio of the green tea albumen of group 1 is significantly higher than group 2, shows that enzymolysis carries in operation afterwards infeasible at alkali.
Embodiment
6
The optimization of enzymolysis process condition in enzyme alkali compound collecting method
1, in Enzymatic Extraction tea leaf protein technique, principal element influential on extraction effect typically has enzyme concentration, pH value, Extracting temperature, extraction time, and for composite hydrolytic enzyme Viscozyme L, this enzyme is 3.3～3.4 at pH value, and under the conditions of temperature is 40 DEG C～50 DEG C, activity is the highest.Through verification experimental verification, optimal pH is 3.4, and optimum temperature is 45 DEG C.
2, enzyme is carried pH value and is set to 3.4 by this experiment, and enzyme temperature raising degree is 45 DEG C, is optimized different enzyme concentrations with extraction time and probes into, determines the optimum extraction process of composite hydrolytic enzyme Viscozyme L extraction tea leaf protein.
(1) optimization of enzyme concentration in enzymolysis step
At temperature 45 C, pH=3.4, in the case of enzyme carries time 1h, selecting enzyme concentration respectively is 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5% to test as extraction conditions, enzyme carry after again alkali carry, during alkali carries, concentration of lye is unified is 0.1mol/L, and temperature is 90 DEG C, and solid-to-liquid ratio is 1:50.
ResultSuch as figure 4Shown in, along with the rising of enzyme concentration, extraction ratio is risen to 72.98% by 53.18%.When adding enzyme concentration less than 2 %, tea protein extracting ratio changes and inconspicuous along with the increase adding enzyme concentration, and when enzyme concentration increases to 2.5% from 2%, the change of tea protein extracting ratio is the most obvious, it is seen that in the case of Gai, the shell-broken effect of plant hydrolyzed-enzyme be the most preferable.And after enzyme concentration reaches 2.5%, curve tends towards stability.Enzyme, as a kind of catalysts, improves reaction efficiency, therefore by being combined performance catalytic action with substrate specificity, when all substrates are all combined with enzyme, catalytic efficiency reaches the highest, if now continuing to improve enzyme concentration, catalytic efficiency there will not be and is obviously improved.Be may determine that optimum extraction enzyme concentration is 3% by curve.
(2) optimization of enzymolysis time in enzymolysis step
At temperature 45 C, pH=3.4, in the case of enzyme concentration 3.0%, select enzyme to carry the time and be respectively 1h, 1.5h, 2h, 2.5h, 3h, 3.5h, test as extraction conditions, enzyme carry after again alkali carry, during alkali carries, concentration of lye is unified is 0.1mol/L, and temperature is 90 DEG C, and solid-to-liquid ratio is 1:50.
ResultSuch as figure 5Shown in, along with enzyme proposes the prolongation of time, extraction rate of protein is totally in slow ascendant trend, and enzyme is held essentially constant after carrying time 3h.This is because in an experiment, enzyme concentration reaches 2.0%, and under this concentration, the shell-broken effect of enzyme is obvious not enough, and when extraction time is 1h, plant cell wall is the most well cracked, and therefore extraction ratio is the lowest.But along with enzyme proposes the prolongation of time, cell wall can be promoted to crack further, the beneficially alkali liquor extraction to protein, therefore extraction ratio raises.When enzyme carries after the time reaches 3h, and extraction ratio is held essentially constant and is because the prolongation along with extraction time, and enzyme activity will decline, and moreover, along with the raising of hydrolysate concentration, enzyme digestion reaction speed slows down, and even there will be reaction and inversely carries out.Therefore, considering various factors, primarily determining that optimal enzyme carries the time is 3h.
Embodiment
7
In enzyme alkali compound collecting method, alkali proposes the optimization of process conditions
1, the optimization of concentration of lye during alkali puies forward step
Being 60min in the fixed extraction time, solid-to-liquid ratio (w/v) is 1:40, under conditions of Extracting temperature is 70 DEG C, to select concentration respectively be 0.04,0.06,0.08,0.10, the NaOH solution of 0.12mol/L tests as extractant.
ResultSuch as figure 6Shown in, the extraction ratio of tea egg white matter raises in the trend gradually risen along with NaOH concentration.When NaOH concentration is 0.02mol/L, tea protein extracting ratio is 13.0%, and when NaOH concentration is 0.12mol/L, tea protein extracting ratio reaches 36.0%, and along with the continuous increase of NaOH concentration, the extraction ratio of tea leaf protein the most constantly increases.After NaOH concentration reaches 0.08mol/L, the rate of rise of tea leaf protein extraction ratio gradually slows down, and has arrived NaOH concentration when being 0.12mol/L, and extraction ratio is held essentially constant.The harmful effect caused in view of tea leaf protein quality and the environmental protection of excessive concentrations NaOH solution pair, finally chooses NaOH concentration 0.10mol/L.
2, the optimization of solid-to-liquid ratio during alkali puies forward step
Being 60min in the fixed extraction time, concentration of lye is 0.1mol/L, under conditions of Extracting temperature is 70 DEG C, tests respectively under the extraction conditions of different solid ratio.
ResultSuch as figure 7Shown in, tea leaf protein extraction ratio is affected relatively big by solid-to-liquid ratio, along with the raising tea protein extracting ratio extracting solid-to-liquid ratio is gradually increased, is risen to 40% by 13%.During alkali carries, when solid-to-liquid ratio is less than normal, the viscosity of solution increases, stirring difficulty, affects the carrying out of mass transport process, and solid-to-liquid ratio can cause the waste of alkali liquor time bigger, and the burden of subsequent treatment, and solid-to-liquid ratio at 1:50 to 1:60 time, extraction ratio raise the slowest, when solid-to-liquid ratio is 1:50, extraction ratio can reach 38.5%.To sum up reason, finally determines that 1:50 solid-to-liquid ratio is tea protein extraction optimal proportion.
3, the optimization of Extracting temperature during alkali puies forward step
Being 60min in the fixed extraction time, solid-to-liquid ratio (w/v) is 1:50, under conditions of concentration of lye is 0.1mol/L, selects temperature to be 50,60,70,80,90 DEG C respectively and tests as extraction conditions.
ResultSuch as figure 8Shown in, in 50 DEG C to 80 DEG C intervals, along with the rising of temperature, tea protein extracting ratio rises more slow, and during to 90 DEG C, extraction ratio is risen to 41% by 34%, is held essentially constant when 100 DEG C afterwards.Tea leaf protein extraction ratio increases along with the rising of temperature.It is likely to result in the protein-denatured problem of tea in view of temperature is the highest, on the premise of not affecting tea protein active, obtains higher extracted rate, determine that Extracting temperature is 90 DEG C.
4, the optimization of extraction time during alkali puies forward step
Being 1:50 in fixing solid-to-liquid ratio (w/v), Extracting temperature is 90 DEG C, under conditions of concentration of lye is 0.1mol/L, to select the time respectively be 20,40,60,80,100min tests as extraction conditions.
ResultSuch as figure 9Shown in, tea leaf protein extraction ratio slowly raises along with the prolongation of extraction time, and after alkali carries 100min, extraction ratio is up to 38%, after alkali carries 120min, extraction ratio is held essentially constant, it is contemplated that time and the situation of energy conservation, the final optimum extraction time selecting 100min to carry as alkali.
Embodiment
8
The optimization of the heavy process conditions of acid in enzyme alkali compound collecting method
1, the optimization of pH in the heavy step of acid
It is that 2.0,2.5,3.0,3.5,4.0,4.5,5.0,5.5 pH value sinking in step to acid are optimized that this experiment is respectively adopted pH, resultTable 3Shown in:
The different pH impact on protein precipitation of table 3Table 3
| pH | 2.0 | 2.5 | 3.0 | 3.5 |
| Rate of deposition (%) | 37.51±0.23 | 37.83±0.07 | 37.57±0.65 | 36.13±0.09 |
| Purity (%) | 54.75±0.81 | 55.35±0.40 | 55.71±0.28 | 54.85±0.18 |
| pH | 4.0 | 4.5 | 5.0 | 5.5 |
| Rate of deposition (%) | 33.73±0.21 | 14.39±0.12 | 0.66±0.16 | 0.40±0.04 |
| Purity (%) | 53.98±1.10 | 51.30±0.27 | 50.97±0.64 | 49.59±0.51 |
FromUpper tableResult show, from the point of view of rate of deposition, pH=2.5 rate of deposition is the highest, and sedimentation effect is optimal；From purity, the tea purity of protein that different pH value are precipitated out is basically unchanged.
It addition, in experimentation, from color and luster, along with the rising of pH value, the color and luster of extracting solution of protein is the most dimmed.This tea polyphenols material Oxidative demage at low ph values being possibly due to residual forms dark matter and precipitates.The most deeply later stage decolouring link may be caused the factors such as trouble in view of the precipitation environment of the peracid protein color and luster that the activity of tea albumen may be impacted and precipitate, use pH=3.0 as the heavy pH value of optimal acid of tea albumen.
2 it addition, sink the optimum results of time of repose of the kind using acid in step and show to acid, and optimal acid is dilute hydrochloric acid, and optimal time of repose is about 30min.
Claims (10)
1. the extracting method of albumen in a green tea tea grounds, it is characterised in that comprise the steps:
S1. enzymolysis: use zymolysis technique that green tea tea grounds is carried out enzymolysis broken wall treatment；
S2. alkali carries: the green tea tea grounds after enzymolysis continues to use alkalinity extraction green tea albumen, obtains green tea protein extract；
S3. acid is heavy: S2 gained green tea protein extract carries out acid heavy, obtains precipitate；
S4. desalination: S3 gained precipitate is carried out desalination；
S5. decolouring: take the green tea albumen after S4 desalination, utilize 75% acetone to decolour, obtain green tea albumen.
The extracting method of albumen in green tea tea grounds the most according to claim 1, it is characterised in that the enzyme that enzymolysis described in step S1 is used is composite plant hydrolytic enzyme.
The extracting method of albumen in green tea tea grounds the most according to claim 2, it is characterised in that described composite plant hydrolytic enzyme is that Novi believes composite plant hydrolytic enzyme Viscozyme L.
The extracting method of albumen in green tea tea grounds the most according to claim 1, it is characterised in that the parameter of enzymolysis described in step S1 is: enzyme concentration is 2.0～3.0%, solid-to-liquid ratio is 1:15～1:30, enzymolysis pH is 3.0～4.0, and hydrolysis temperature is 40～50 DEG C, and enzymolysis time is 1～3h.
The extracting method of albumen in green tea tea grounds the most according to claim 1, it is characterized in that, the parameter that alkali described in step S2 carries is: concentration of lye is 0.08～0.12mol/L, and solid-to-liquid ratio is 1:40～1:60, Extracting temperature is 70～90 DEG C, and extraction time is 60～120min；It is centrifuged and obtains green tea protein extract.
The most according to claim 1, the extracting method of albumen in green tea tea grounds, it is characterised in that described in step S3, acid is heavy is to take green tea protein extract, be 2～3 with dilute hydrochloric acid regulation pH, centrifugal after standing 20～40min, is precipitated thing.
The extracting method of albumen in green tea tea grounds the most according to claim 1, it is characterized in that, desalination described in step S4 is to add water and mix in precipitate, inject in bag filter, it is placed in pure water dialysis 24～48h, period changes pure water 3～5 times, until the outer water colorless of bag filter and electrical conductivity are close to pure water, then 3000～4000r/min are centrifuged 10～20min and obtain the green tea albumen after desalination.
The extracting method of albumen in green tea tea grounds the most according to claim 1, it is characterised in that green tea tea grounds described in step S1 is the discarded green tea tea grounds after extraction.
The extracting method of albumen in green tea tea grounds the most according to claim 1, it is characterised in that green tea albumen and the amount ratio of 75% acetone after desalination described in step S5 be: solid-to-liquid ratio 1:4～1:8.
The extracting method of albumen in green tea tea grounds the most according to claim 1, it is characterized in that, described in step S5, the method for decolouring is: take the green tea albumen after S4 desalination, add 75% acetone, shake 10～20min, be centrifuged and abandon supernatant, it is repeated several times, with pure water and be centrifuged, it is repeated several times with clean acetone, obtains green tea albumen.
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