CN109619233A - A kind of preparation method of anti-oxidation function instant tea powder - Google Patents
A kind of preparation method of anti-oxidation function instant tea powder Download PDFInfo
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23F—COFFEE; TEA; THEIR SUBSTITUTES; MANUFACTURE, PREPARATION, OR INFUSION THEREOF
- A23F3/00—Tea; Tea substitutes; Preparations thereof
- A23F3/16—Tea extraction; Tea extracts; Treating tea extract; Making instant tea
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23F—COFFEE; TEA; THEIR SUBSTITUTES; MANUFACTURE, PREPARATION, OR INFUSION THEREOF
- A23F3/00—Tea; Tea substitutes; Preparations thereof
- A23F3/16—Tea extraction; Tea extracts; Treating tea extract; Making instant tea
- A23F3/30—Further treatment of dried tea extract; Preparations produced thereby, e.g. instant tea
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Abstract
The invention belongs to functional food production technical fields, and in particular to a kind of preparation method of anti-oxidation function instant tea powder, product contain the several functions compositions such as tea polyphenols, tea polypeptide, tea polysaccharide.Preparation method is to assist lywallzyme and protease difference enzymolysis and extraction, filtering, centrifugation, homogeneous, spray drying that water-soluble preferable anti-oxidant dry tea powder is made through low power ultrasound after crushing tea raw material.Raw material of the present invention enzymolysis and extraction at low temperature, extracting solution high-temperature instantaneous is dry, preferably save antioxidation biology activity, the anti-oxidation function instant tea powder antioxidant activity of preparation is high, physical behavior is good, it can be used as functional food additives use, be that raw material can filling capsule or tabletting production functional food.
Description
Technical field
The invention belongs to functional food production technical fields, and in particular to a kind of preparation side of anti-oxidation function instant tea powder
Method.
Background technique
Tealeaves is the important beverage in people's daily life, have drink, nutrition, health care, pharmacology, the polynary valence such as humanity
Value.2016,2,870,000 hectares of Chinese tea place area, more than 240 ten thousand tons of yield, consumption figure was more than 2,000,000 tons, it has also become the whole world is most
Big Chan Cha state and Tea Consumption market.A large amount of tea stalk, tea dust, tea grounds etc. can be generated in tealeaves and its product process
Leftover bits and pieces and inferior matter tealeaves, turn waste into wealth, and develop and use to these leftover bits and pieces and inferior matter tealeaves, not only can increase tea processing factory's warp
Ji benefit, and there is the preferable ecological value.
Contain several physiological active substances, including tea leaf protein, tea polyphenols, active polysaccharide, theafolisaponin, coffee in tealeaves
The plurality of active ingredients such as alkali, vitamin and Fe, Se, γ-aminobutyric acid, theanine.
Protein is the important nutriment of people.Tea egg white matter (tea protein) account for about dry weight of tea leaves 15%~
30%, mainly include glutelin (82.05%), alcohol soluble protein (13.61%), albumin (3.47%), globulin (0.87%),
Containing 16~18 kinds of amino acid, wherein aspartic acid, glutamic acid, serine, alanine, threonine and Glycine Levels are higher.
Changhai hospital, The 2nd Army Medical College Nutriology Dept. professor Cai Donglian has done a large amount of experiment to animal and human body, experiments have shown that tealeaves
Healthcare function possessed by protein is no less than tea polyphenols, finishes the classical opinion that tea egg white matter only belongs to trophic function from this
It is disconnected.Li Yan, Xiao of Changhai hospital make equality and find in the research for carrying out the prevention radiation of tea egg white matter with mouse, tea leaf protein
Matter has the function of removing superoxide anion, can resist mutagenic effect caused by ionising radiation.Yao Huiying etc. reports tea egg
White matter also plays the role of removing free radical, and is also eager to excel than the ability of tea polyphenols.The Hu Chang of Wuxi Light Industry Univ.'s Foodstuffs Academy
Cloud, which is also studied, show that tea egg white matter has oxidation resistant ability.
Tea polyphenols (tea polyphenols abbreviation TP) are the general names of a variety of phenols and its derivative in tealeaves, in tealeaves
In account for 14~24%.Tea polyphenols have extremely strong antioxidation and various health care functions, to superoxide anion radical
With effect and extremely strong antioxidant activity is eliminated very much by force, people's Antioxidation can be inhibited.China was in 1991
Food national standard is included in using tea polyphenols as a kind of natural.Tea polyphenols are to staphylococcus, Escherichia coli, tangerine grass
Nearly hundred kinds of bacteriums have excellent antibacterial activity in the natures such as bacterium, bacillus, golden yellow streptococcus, and also having reduces body
The effect of rouge, liver rouge quantity reduces blood pressure and hinders the effect for delaying coagulating platelets, and atherosclerosis prevents cardiovascular
A variety of pharmacological functions such as disease.Largely studies have shown that tea polyphenols in vitro have antimutagenic effect, rodent can be inhibited
The generation of kinds of tumors improves the immunity of body, and the metabolic pathway for blocking carcinogen to be formed inhibits tumour cell deoxidation core
The biosynthesis of ribosomal ribonucleic acid.
Tea polysaccharide (tea polysaccharide abbreviation TPS) is a kind of and combined acidity of protein in tealeaves
Polysaccharide or a kind of acidoglycoprotein, content is 2%~7%, and general green tea is higher than black tea, by arabinose, xylose, rock algae
Sugar, glucose, galactolipin etc. are constituted.Pharmacological research shows that tea polysaccharide has hypoglycemic, prevention and treatment diabetes, reducing blood lipid, anticoagulant
Blood, antithrombotic, blood pressure lowering, reducing heart rate, resist oxygen lack and the medicines such as the effect of increase coronary artery flowing and anti-inflammatory, radiation protection
Reason effect, moreover it is possible to increase serum agglutinin antibody, to enhance body's immunity, tea polysaccharide also has NaNO2 very strong
Scavenging effect.
Caffeine (caffeine) is the main substance of tealeaves alkaloid, and basic structure is by a pyrimidine ring and one
1,3,7- trimethyl xanthine made of imidazole ring is condensed.Caffeine content is 2%~5% in tealeaves, the tea making such as tea dust, tea stalk
Content is 2.5% or so in waste material.Caffeine is not only to constitute the important component of millet paste flavour, and has heart tonifying, diuresis, solution
Poison, the pharmacological functions such as relieving asthma, can be used for the treatment that bronchus is wheezed, and clinically also treat heart failure using caffeine.Coffee
Alkali also has the function of promoting blood circulation, and vascular smooth muscle can be made loose, increases the effective diameter of blood vessel, enhances painstaking effort tube wall
Elasticity has positive blockage effect to heart, thus has the function of refresh oneself benefit think of, hypoglycemic, blood lipid, blood pressure.
Theafolisaponin (tea saponin) is a kind of more complex glucoside derivative, belongs to the olive in triterpenoid saponin
Alkane type, basic structure are made of aglucon and sugared body two parts.Theafolisaponin has the function of reduction cholesterol, prevents gallbladder solid
Absorption of the alcohol in enteron aisle, to reduce the cholesterol in blood plasma lipide.Theafolisaponin can be in conjunction with macromolecular alcohol such as sterols
It forms double salt and shows haemocylolysis, there is antibacterium and antimycotic activity, also have and kill mollusk, anti-inflammatory, calm
Isoreactivity.
In addition, also a variety of effectively containing vitamin and F, Fe, Se, γ-aminobutyric acid, theanine, Tea Pigment etc. in tealeaves
Ingredient, they have certain health-care efficacy to human body.China's most area belongs to low fluorine zone, supplements suitable fluorine to human body
Sclerous tissues' (skeleton, tooth) constitute and are advantageous, especially to Common Pediatric Diseases saprodontia and old common disease skeleton osteoporosis
Prevention and treatment is advantageous；γ-aminobutyric acid is for increasing the activity of glucose phosphate Esterified Enzyme, reducing blood pressure and the content of ammonia in blood, extensive
Multiple function of brain cell, treatment hepatic encephalopathy have obvious action.
Tealeaves functional components can be divided into two classes according to content, and a kind of content is obviously more, 20% or so, cry largely at
Point, remaining content is less, generally below 10%.A large amount of ingredients have tea protein and tea polyphenols, are the main of tealeaves extraction
The bulk composition of target and tealeaves functionality extract, therefore the present invention is also detection other side with both ingredients, to evaluate
Extract.By water solubility, the functional components in tealeaves are divided into ease of solubility ingredient and refractory components two major classes.Tea polyphenols, tea
The functional components such as polysaccharide, caffeine, theanine are soluble easily in water, and separation and Extraction is easier to.Tea egg white matter is not readily dissolved in water, belongs to
Refractory components.Water-soluble portion accounts for about the 1%~2% of total amount in tea egg white matter, and the overwhelming majority is water-insoluble.Therefore,
Tea egg white matter is as the highest functional components of content, and few due to being not readily dissolved in water content in tea juice, the overwhelming majority deposits
It is in tea grounds, is abandoned with tea grounds, causes the serious wasting of resources.In order to separate and extract tea egg white matter from tea grounds,
Chen Shixue etc. has studied microwave radiation exaraction tea egg white matter technique, and Shen Lian is clear etc. has studied the alkalinity extraction of protein in tea grounds
Technique improves the recovery rate of tea egg white matter by Optimized Extraction Process, but the tea egg white matter dissolubility extracted is not
It changes.Tea egg white matter is still not easy to utilize, and the extremely low status of utilization rate is still without change.
The dissolubility of protein is considered as most important aspect in functional character, affects other functionality of protein
Matter, good dissolubility can expand the potential application of protein.Since most tea egg white matters are not soluble in water, it is difficult to extract benefit
With, therefore to be suitably modified.Enzymatic hydrolysis is a kind of protein-modified important method, and insoluble tea egg white matter is through protease enzyme
Significant change has occurred in Xie Hou, property.Modified protein molecule is degraded, and peptide chain length shortens, the carboxylic exposed
Base, amino, hydroxyl are more, and polarity enhancing so that modified tea egg white matter dissolubility be made to be remarkably reinforced, is conducive to mention
It takes.Moreover, tea egg white matter can generate some functional peptides during enzymatic hydrolysis, antioxidant activity is shown, can be used for function
Energy property food production is used as functional food additives.
With the improvement of people ' s living standards with the enhancing of health care consciousness, the demand of functional food can increase year by year.Mesh
Before, the healthcare function of tealeaves is generally approved by masses, and exploitation tealeaves functional food can comply with the market demand, easily by consumer
Receive, there are good market prospects.The present invention provides a kind of new method, tealeaves or tea processing leftover bits and pieces by surpassing twice
Sound assistance enzymolysis extracts to prepare functional components.Enzymatic hydrolysis effect is different twice, and enzymatic hydrolysis establishes base for second of enzymatic hydrolysis for the first time
Plinth.Ultrasonic wave added lywallzyme enzymatic hydrolysis can effectively destroy tea cell wall, make tea polyphenols in tea cell, tea polysaccharide, caffeine etc.
Soluble functional ingredient Fast Stripping, high efficiency extraction come out.The substances such as tea polyphenols, caffeine generate easily in conjunction with protein
Precipitating, proteolytic is advantageous when extracting after extraction to second；Meanwhile enzymatic hydrolysis destroys cell complete structure, makes slightly solubility
The exposure of tea egg white matter, protease hydrolyzed when conveniently extracting for the second time.Ultrasonic wave added protease hydrolyzed can high effective and modified tea egg
White matter makes part Investigation of Hydrolytic Cleavage of Peptides, is converted to functional peptide soluble and with antioxidant activity.And due to molecular weight
Become smaller, is not easy to generate precipitating in conjunction with the substances such as tea polyphenols, therefore solution of extract stability enhances.Digest purpose not twice
Together, it complements one another, sequence is immutable, can not be merged into primary enzymolysis.By the anti-oxidant soluble matter for extracting preparation twice
Matter is based on tea polyphenols and small peptide, while the tealeaves soluble functional ingredient containing there are many, is a kind of good functional food
Or functional food additives.Tea resources waste can be reduced using the present invention, reduces tea processing factory's production cost, increases tealeaves and produces
Value alleviates tea processing waste to the pressure of environment.
Summary of the invention
The present invention is insufficient in the prior art in order to solve, and provides a kind of preparation side of anti-oxidation function instant tea powder
Method, the present invention destroys cell wall using the compound lywallzyme of ultrasonic wave added and extracts ease of solubility antioxidant content, then auxiliary with ultrasound
Help compound protease enzymatic hydrolysis tea egg white matter that tealeaves anti-oxidation peptide extracting solution is made, this can be obtained in spray drying after merging filtrate
Anti-oxidation function instant tea powder in invention.The present invention is using antioxidant activity as index optimization extraction process, the prominent function of extract
It can be antioxidant activity, main component is tea polyphenols, tea polypeptide, but also containing other some soluble functional ingredients, therefore
Function is not limited solely to anti-oxidant.Extract others functional activity, can refer to the function of tealeaves, due to not examining specifically
It surveys, therefore range is not discussed in the present invention.
The technical solution adopted by the present invention to solve the technical problems is:
A kind of preparation method of anti-oxidation function instant tea powder, described method includes following steps:
S1, it takes tealeaves or/and tea comprehensive processing leftover bits and pieces to smash it through 40~60 meshes, obtains tea powder, tea powder is placed in can
In temperature controlled ultrasonic extractor, add distilled water immersion 10~30 minutes, the mass ratio of tea powder and distilled water is 1:(10~60)；
S2, it is added compound lywallzyme in the soak of step S1, ultrasonic wave added enzymolysis and extraction filters after enzymatic hydrolysis,
Filtrate and filter residue difference are spare；
S3, distilled water and compound protease are added into the filter residue of step S2, ultrasonic wave added enzymolysis and extraction, after enzymatic hydrolysis
Heat up enzyme deactivation, filtering, and filtrate is merged to obtain extracting solution with the resulting filtrate of step S2；
S4, by extracting solution centrifugation, filter, after high-pressure homogeneous, spray drying up to anti-oxidation function instant tea powder.
Tea raw material includes tea stalk, tea dust, the tea head etc. of generation during tea processing, further includes unsalable coarse tea,
These source commodities are poor, and generally as offal treatment, tea raw material is dry, crushed with beater grinder, grinding and sieving,
The part of 40~60 mesh of particle diameter is taken to use.
The mass ratio of the tea raw material powder and water is 1:10~60, and tea raw material powder and water total volume are that extractor is overall
Long-pending 80%~95%.
Preferably, the compound lywallzyme is in pectase, cellulase, hemicellulase, driselase and glusulase
At least three kinds it is compound；The compound protease is papain, alkali protease, neutral proteinase, bromelain, wind
At least two in taste protease and mushroom enzyme are compound.
Preferably, the compound lywallzyme is that pectase, cellulase and glusulase are compound, the pectase, fiber
The compound ratio of plain enzyme and glusulase is 100~200U:150~300U:60~100U；The compound protease is bromelain
Compound with alkali protease, the compound ratio of the bromelain and alkali protease is 100~200U:180~350U.
Preferably, the enzyme concentration of compound lywallzyme is the 0.5%~3.5% of tea powder weight in step S2；In step S3
The enzyme concentration of compound protease is the 1%~4% of tea powder weight.
S2 and S3 are extracted as same container twice, are controllable temperature ultrasonic extraction tank, and ultrasound used is low powder pulsed ultrasound,
Ultrasonic power is 30~120W in step S2, and the ultrasound works time is 3~5s, and the intermittent time is 1~3s；Ultrasonic function in step S3
Rate is 50~150W, and the ultrasound works time is 2~5s, and the intermittent time is 1~4s.
Preferably, pH is 5.5~8.0 in step S2, temperature is 20 DEG C~60 DEG C, and enzymolysis time is 10~120min；
PH is 6.5~9.0 in step S3, and temperature is 30 DEG C~70 DEG C, and enzymolysis time is 20~120min.
Preferably, pH is 6.0~7.5 in step S2, temperature is 30 DEG C~50 DEG C, and enzymolysis time is 20~60min；Step
PH is 6.5~8.0 in rapid S3, and temperature is 40 DEG C~60 DEG C, and enzymolysis time is 30~80min.
Preferably, the temperature of the heating enzyme deactivation in step S3 is 90 DEG C~95 DEG C, the enzyme deactivation time is 1~5min.
Preferably, in step S4 extracting solution through revolving speed be 6000~8000r/min centrifuge be centrifuged 10~20min,
Remove precipitating, 200 mesh filter cloth suction filterings of clear liquid, high-pressure homogeneous pressure be 15MP~30MP, homogenisation cycle 5~15 times.
Preferably, the inlet air temperature being spray-dried in step S4 is 160 DEG C~200 DEG C, leaving air temp is 80 DEG C~100
℃。
The screening process of main technologic parameters of the invention described further below:
In order to guarantee the accuracy of experiment, keep experimental raw with uniformity, the present invention carries out various process parameters optimization
The raw materials used single tea powder provided for complete ecological tea processing factory, Chaozhou man is tested, this tea powder is to lose quotient during tea processing factory produces tealeaves
The broken tea of moral character, particle size is different, is crushed with beater grinder, crosses 40 meshes.Tea grounds powder used is made for single tea powder,
Preparation method: tea powder adds 20 times of quality distilled water, and 100 DEG C of extraction 30min repeat extraction 2 times, filtering, and filter residue is dried for standby.
Choice of parameters is to remove DPPH ability as index using extracting solution.DPPH (1,1- diphenyl -2- picryl phenylhydrazine) is
A kind of stable free radical centered on nitrogen, clearance rate height reflect the power of extracting solution antioxidant activity, extracting solution
It is as follows to DPPH clearance rate measuring method:
The DPPH solution for being 0.04mg/mL with dehydrated alcohol compound concentration is added 2mL into 2mL DPPH solution and extracts
Liquid mixes, and stands 30min, is centrifuged (3000r/min, 10min), supernatant is taken to survey light absorption value at 517nm.
The ability that extracting solution removes DPPH indicates that clearance rate is bigger with clearance rate, and the ability for removing DPPH is stronger.It is public
Formula are as follows:
In formula, SE is clearance rate；Ai is 2mL DPPH solution+2mL extracting solution absorbance value；Aj be 2mL dehydrated alcohol+
2mL extracting solution absorbance value；A0 is 2mL DPPH solution+2mL water absorbance value.
1, combined-enzyme method extract bioactive substance have been developed in recent years a kind of new technology, it for tea polyphenols,
The cell area of tea polysaccharide isoreactivity substance distribution is different, using different compound enzyme systems, under the synergistic effect of various enzymes,
It is sufficiently destroyed eucaryotic cell structure, improves the yield of bioactive substance to the maximum extent, while keeping its inherent structure and activity.It should
Method is applied in tea polyphenols, polysaccharide, protein extraction, and obtains preferable effect.Combined-enzyme method extracts soluble component
Professional equipment is not needed, industrial amplification is easy to, it is environmental-friendly, and as biologic product technology develops, enzyme price further drops
Low, extraction cost can further decrease, with good application prospect.
The present invention extracts tealeaves active material using combined-enzyme method, and it is as follows to optimize compound enzyme test: accurately weighing
5 parts of 0.5000g tea powder, it is separately added into 10mL distilled water, impregnates 30min, 40 DEG C of waters bath with thermostatic control add respectively into 5 parts of tea powders and collapse
It bursts enzyme, glusulase, cellulase, pectase, each 2000U of hemicellulase, ultrasound enzymatic hydrolysis 30min, supersonic frequency under 60W power
40kHz；In 95 DEG C of heating 3min enzyme deactivations after enzymatic hydrolysis, tap water is cooling, and filtrate scavenging activity on DPPH is surveyed in filtering, and DPPH is clear
Show that the anti-oxidation active substance of dissolution is more except rate is higher.Every kind of enzyme sets 3 repetitions, calculates average value, and driselase, snail is added
Ox enzyme, cellulase, pectase, hemicellulase DPPH clearance rate be respectively as follows: 30.74%, 37.82%, 42.48%,
40.18%, 27.91%.Clearance rate height indicate anti-oxidation active substance content in extracting solution number, can be with from clearance rate
Find out that the extraction effect of cellulase, pectase and glusulase is substantially better than driselase and hemicellulase, so choosing fiber
Plain enzyme, pectase and glusulase are compound.The present invention is in bibliography (Xu Jie etc., the technical study of enzyme process assisted extraction tea polyphenols)
And on the basis of a large amount of composite effect tests, comprehensively considers the height of three kinds of enzyme commodity prices, determine pectase: cellulase:
Glusulase=100~200U:150~300U:60~100U, tealeaves antioxidant content recovery rate is higher under the compound condition, and
Lower production costs have higher practical value.
Protein becomes the lesser small peptide of molecular weight by protease hydrolytic, polypeptide chain degradation, and internal group exposure is simultaneously formed
Specific space structure, to generate certain bioactivity.Proteolytic enzyme protolysate matter has certain specificity, i.e., to urging
Changing peptide bond has selectivity.After protease is compound, different protease hydrolytic peptide bonds are different, can produce synergistic effect, imitate hydrolysis
Rate greatly improves.
The present invention prepares anti-oxidation peptide using compound protease, and it is as follows to optimize compound enzyme test: accurately weighing 1g tea grounds
4 parts of powder, respectively plus distilled water 50mL impregnates 30min, is separately added into bromelain, papain, neutrality into 4 parts of tea grounds powder
Each 400U of protease, alkali protease adjusts pH value in the most suitable range of each protease, is respectively as follows: 7,7,7,8.Hydrolysis temperature is
The lower limit of each enzyme optimum temperature range is respectively as follows: 55 DEG C, 50 DEG C, 40 DEG C, 35 DEG C.Ultrasound enzymatic hydrolysis 30min, is extracted under 60W power
After 95 DEG C of heating 3min enzyme deactivations, enzymolysis liquid filtering, survey filtrate scavenging activity on DPPH, test set 3 repetitions, be averaged,
Be added bromelain, papain, neutral proteinase, alkali protease the clearance rate of enzymolysis liquid be respectively as follows:
57.01%, 36.17%, 27.89%, 66.51%.From clearance rate it can be seen that the enzymatic hydrolysis of bromelain and alkali protease
Effect is substantially better than papain and neutral proteinase, so choosing bromelain and alkali protease as complex enzyme group
Point.The present invention comprehensively considers the height of two kinds of enzyme commodity prices on the basis of a large amount of compound proportioning tests, finally determines
Compound proportion is bromelain: alkali protease=100~200U:180~350U.It digests and obtains under the compound conditions of mixture ratios
Tealeaves antioxygen peptide activity it is higher.
2, for the enzymatic reaction system in enzymolysis process, as enzyme concentration increases, enzyme-to-substrate touch opportunity increases, together
The molecular number of enzyme hydrolysis increases in one time, and soluble component is caused quickly to separate.When enzyme concentration is increased to certain journey
Degree, enzyme molecule are excessively saturated, and part enzyme molecule is had no chance and Binding Capacity, and the speed that substrate is digested is not further added by, therefore
Excessive enzyme can not accelerate extraction rate and anti-oxidation peptide yield.Complex enzyme additive amount Optimum Experiment of the invention is as follows:
5 parts of 0.5000g tea powder accurately are weighed, it is each that 10mL distilled water is added, impregnate 30min.Tea is separately added into tea powder
The compound lywallzyme of powder 0.5wt%, 1wt%, 1.5wt%, 2wt%, 2.3wt%, 3wt%, 3.5wt%, 4wt%, 40 DEG C,
Ultrasound enzymatic hydrolysis 20min under pH value 7,60W power.95 DEG C of heating 3min enzyme deactivations, tap water are cooled to room temperature after extraction, mistake
Filtrate scavenging activity on DPPH is surveyed in filter, and test sets 3 repetitions, is averaged, and the highest enzyme concentration of extracting solution DPPH clearance rate is most
Good enzyme concentration.
5 parts of 1.0000g tea grounds powder accurately is weighed, respectively plus distilled water 50mL, impregnates 30min.It is separately added into tea grounds powder
The compound protease of tea grounds powder 0.5wt%, 1wt%, 1.5wt%, 2wt%, 2.3wt%, 3wt%, 3.5wt%, 4wt%, 50
DEG C, ultrasound enzymatic hydrolysis 20min under pH value 7,60W power.95 DEG C of heating 3min enzyme deactivations, tap water are cooled to room temperature after extraction,
Filtering, surveys filtrate scavenging activity on DPPH, and test sets 3 repetitions, is averaged, the highest enzyme concentration of extracting solution DPPH clearance rate is
Best enzyme concentration.
3, pH influences enzyme activity.PH value appropriate maintains the best three-dimensional conformation of enzyme active center by electrostatic interaction,
Enzyme-to-substrate is promoted to combine.PH also will affect enzymatic speed, and pH value is different, the electrically charged difference of enzyme and substrate specificity institute, enzyme
The affinity of three-dimensional conformation and enzyme-to-substrate is different, and Catalysis Rate is also just different.PH value Optimum Experiment of the invention is as follows:
5 parts of 0.5000g tea powder accurately are weighed, it is each that 10mL distilled water is added, 30min is impregnated, the pH of each solution is adjusted separately
Value is 5,5.5,6,6.5,7,7.5,8,8.5,9, at 40 DEG C of temperature, under conditions of adding compound lywallzyme amount to be tea powder 1.5wt%
With 60W power ultrasound enzymolysis and extraction 20min, pH meter online detection and control pH value is used in extraction process.Add for 95 DEG C after extraction
Hot 3min enzyme deactivation, tap water are cooled to room temperature, filtering, survey filtrate scavenging activity on DPPH, and test sets 3 repetitions, is averaged, mentions
Taking liquid to remove the highest pH value of DPPH rate is optimal pH.
5 parts of 1.0000g tea grounds powder accurately is weighed, respectively plus distilled water 50mL, immersion 30min, compound protease enzyme concentration are
The 2wt% of tea grounds powder, adjusting pH value is respectively 6.5,7,7.5,8.0,8.5,9.0,50 DEG C, ultrasound enzymatic hydrolysis 20min under 60W power,
95 DEG C of heating 3min enzyme deactivations, tap water are cooled to room temperature after extraction, filtering, survey filtrate scavenging activity on DPPH, and test is set 3 times
It repeats, is averaged, the highest pH value of extracting solution DPPH clearance rate is optimal pH.
4, in enzymic catalytic reaction (i.e. enzymolysis and extraction process), temperature is one of important parameter.As the temperature rises, enzyme
The energy of increased activity, reactant increases, and reaction speed is accelerated.After its optimum temperature, zymoprotein mutability, so that enzyme
Vigor weakens, until completely losing, reaction speed can also decline rapidly as the temperature rises.Temperature optimization test of the invention
It is as follows:
5 parts of 0.5000g tea powder accurately are weighed, it is each that 10mL distilled water is added, 30min is impregnated, adjusting pH value is 7.0, adds tea powder
The compound lywallzyme of 1.5wt% uses 60W power ultrasound enzyme under the conditions of 20 DEG C, 30 DEG C, 40 DEG C, 50 DEG C, 60 DEG C of waters bath with thermostatic control
Solution extracts 30min.95 DEG C of heating 3min enzyme deactivations, tap water are cooled to room temperature after extraction, filtering, are surveyed filtrate and are removed DPPH
Activity, test set 3 repetitions, are averaged, the highest temperature of extracting solution DPPH clearance rate is best.
5 parts of 1.0000g tea grounds powder accurately is weighed, respectively plus distilled water 50mL, immersion 30min, tune pH value are 7.5, add tea powder
The compound molten wall-held protein enzyme of 2wt%, with 60W power ultrasound under the conditions of 30 DEG C, 40 DEG C, 50 DEG C, 60 DEG C, 70 DEG C of waters bath with thermostatic control
Enzymolysis and extraction 30min.95 DEG C of heating 3min enzyme deactivations, tap water are cooled to room temperature after extraction, filtering, are surveyed filtrate and are removed
DPPH activity, test set 3 repetitions, are averaged, the highest temperature of extracting solution DPPH clearance rate is best.
5, enzyme digestion reaction time and enzymatic hydrolysis carry out degree have close relationship, and the reaction time is too short, and enzymatic hydrolysis is insufficient, mention
Take rate lower；Reaction time, the too long functional components that not can increase dissolved out, and instead increased impurity dissolution, therefore enzymolysis time
Influence functional components recovery rate and purity.Extraction time Optimum Experiment of the invention is as follows:
5 parts of 0.5000g tea powder accurately are weighed, respectively plus distilled water 10mL, immersion 30min, compound lywallzyme enzyme concentration are tea
The 1.5wt% of ground-slag, hydrolysis temperature are 40 DEG C, and adjusting pH value is that ultrasound enzymatic hydrolysis, enzymolysis time are respectively under 7.0,60W power
10min,20min,30min,40min,50min,60min.95 DEG C of heating 3min enzyme deactivations, are originally water-cooled to room after extraction
Filtrate scavenging activity on DPPH is surveyed in temperature, filtering, and test sets 3 repetitions, is averaged as extracting solution DPPH clearance rate highest
Extraction time be best.
5 parts of 1.0000g tea grounds powder accurately is weighed, respectively plus distilled water 50mL, immersion 30min, compound protease enzyme concentration are
The 2wt% of tea grounds powder, hydrolysis temperature are 50 DEG C, and adjusting pH value is that ultrasound enzymatic hydrolysis, enzymolysis time are respectively under 7.5,60W power
10min,20min,30min,40min,50min,60min.95 DEG C of heating 3min enzyme deactivations, are originally water-cooled to room after extraction
Filtrate scavenging activity on DPPH is surveyed in temperature, filtering, and test sets 3 repetitions, is averaged as extracting solution DPPH clearance rate highest
Extraction time be best.
6, ultrasonic wave is a kind of good directionality, the strong mechanical vibration wave of penetration capacity.When ultrasonic wave is propagated in the medium,
Due to the interaction of ultrasonic wave and medium, make medium that physics and chemical change occur, to generate mechanical effect, cavitation is made
The effects of with, fuel factor.Low frequency ultrasound fuel factor is smaller, and solution heating is unobvious, but can increase mass-transfer efficiency, makes bottom
Object molecule enters the quickening of enzyme active center zone velocity, improves the substrate effective concentration in enzyme active center region, dramatically increases
Enzymolysis speed.Optimization ultrasonic power test of the invention is as follows:
5 parts of 0.5000g tea powder accurately are weighed, it is each that 10mL distilled water is added, 30min is impregnated, adds that tea powder 1.5wt%'s is compound
Lywallzyme, tune pH value be 7.0,40 DEG C of waters bath with thermostatic control, then respectively ultrasonic power 0W, 30W, 45W, 60W, 75W, 90W,
Enzymolysis and extraction 30min under 105W, 120W.95 DEG C of heating 3min enzyme deactivations, tap water are cooled to room temperature after extraction, are filtered, and are surveyed
Filtrate scavenging activity on DPPH, test set 3 repetitions, are averaged, the highest ultrasonic power of extracting solution DPPH clearance rate is
Most preferably.
5 parts of 1.0000g tea grounds powder accurately is weighed, it is each that 50mL distilled water is added, 30min is impregnated, adds that tea powder 2wt%'s is compound
Protease, tune pH value be 7.5,50 DEG C of waters bath with thermostatic control, then respectively ultrasonic power 0W, 30W, 45W, 60W, 75W, 90W,
Enzymolysis and extraction 30min under 105W, 120W.95 DEG C of heating 3min enzyme deactivations, tap water are cooled to room temperature after extraction, are filtered, and are surveyed
Filtrate scavenging activity on DPPH, test set 3 repetitions, are averaged, the highest ultrasonic power of extracting solution DPPH clearance rate is
Most preferably.
The measuring method of peptide, tea polyphenols and water content is as follows in target product anti-oxidation function instant tea powder of the invention:
1) the measurement Forint phenol method of peptide content
A. the preparation of alkaline copper test solution: sodium hydroxide 10g, sodium carbonate 50g is taken to add water 400mL to make to dissolve, as solution A；
Potassium tartrate 0.5g is taken, water 50mL is added to make to dissolve, separately takes copper sulphate 0.25g, water 30mL is added to make to dissolve, two liquid are mixed and are used as second
Liquid.Before use, merge two liquid, and add water to 500mL.
B. standard protein solution: bovine serum albumin is after micro-Kjeldahl measures protein content, according to it
Purity is configured to 0.3mg/ml standard solution with distilled water, with 100ml volumetric flask constant volume.
C. standard curve: precision measures standard protein solution 0.0mL, 0.1mL, 0.3mL, 0.5mL, 0.7mL, 0.9mL,
It sets in tool plug test tube respectively, respectively adds water to 1.0mL, be separately added into alkaline ketone test solution 1.0mL, shake up, it is each that forint phenol test solution is added
4.0mL is mixed immediately, is set accurate response 5min in 55 DEG C of water-baths, is set 10min in cold bath, and extinction is measured at 650nm wavelength
Degree；Simultaneously using No. 0 pipe as blank.Using absorbance as ordinate, protein concentration (mg/ml) is that abscissa draws standard song
Line.Bovine serum albumin concentration 0.030~
Good linear relationship is presented with absorbance value within the scope of 0.270mg/ml.Regression equation are as follows:
Y=2.38576x+0.00539, R2=0.99964
D. dry anti-oxidation function instant tea powder 10mg, is settled to 10mL with distilled water.1mL is taken, by above-mentioned same sample prescription
Method measures the OD value at 540nm, if 3 repetitions, average.Surveyed absorbance brings standard curve into and obtains peptide concentration in sample
C(mg/mL)。
Anti-oxidation function instant tea powder peptide content=C × 100%
2) " the detection side of Tea Polyphenols in Tea and catechin content GB/T 8313-2008 is pressed in polyphenol content measurement
Method " it carries out.
3) measurement of water content
Baking oven is opened in advance, temperature rises to 80 DEG C.It weighs a certain amount of antioxygen functional instant tea powder W (g) and is put into culture
In ware, weighs W1 (g), be then placed in baking oven, 80 DEG C dry to constant weight together with culture dish.It takes out, is put into drier and is cooled to
Room temperature is weighed W2 (g).Water content (%)=(W1-W2)/W × 100% in anti-oxidation function tea powder.
The beneficial effects of the present invention are:
The present invention crushes tea raw material, first extracts soluble antioxidant content with low power ultrasound auxiliary lywallzyme,
It prepared again with low power ultrasound auxilin enzyme, extract antioxidation active peptides, extracting solution is dried by high-temperature instantaneous, is preferably saved
Extract antioxidation biology activity.The anti-oxidation function instant tea powder main component of preparation is soluble tea polyphenols and polypeptide,
Wherein polyphenol content >=32wt%, peptide content >=40wt%, water content≤3.6wt%, 1mg/mL solution D PPH clearance rate
>=18%.The product has preferable instant capacity, can be dissolved in water, solution clear, no precipitating, since product is anti-oxidant rapidly
Active component content is high, physical behavior is good, can be used as food additives use, also can be used as the filling capsule of raw material or tabletting, raw
Produce functional food.
Detailed description of the invention
Fig. 1 is production technological process of the invention.
Specific embodiment
Below by embodiment, in conjunction with attached drawing, explanation, but the embodiment are further described to technical solution of the present invention
It does not limit the scope of the invention.
Embodiment 1:
A kind of preparation method of anti-oxidation function instant tea powder includes the following steps:
S1, it takes 500g tea powder to be placed in controllable temperature ultrasonic extraction tank, adds 20~30L of distilled water, impregnate 30 minutes；
S2, compound lywallzyme ultrasonic wave added enzymolysis and extraction is added in the soak of step S1, compound lywallzyme is by pectin
Enzyme: cellulase: glusulase=200U:300U:100U is combined, enzyme concentration be tea powder dry weight 0.8wt%~
2.5wt%, pH=6~7.5,35~40 DEG C of temperature, 50~120W of ultrasonic power, 3~5s of ultrasound works time, the intermittent time 1
~3s, 20~120min of extraction time are filtered after enzymatic hydrolysis, and filtrate and filter residue difference are spare；
S3,20~25L distilled water is added into the filter residue of step S2, adds compound protease ultrasonic wave added enzymatic hydrolysis and mentions
Take, compound protease is by bromelain: alkali protease=200U:300U is combined, and enzyme concentration is tea powder dry weight
1.0wt%~3.0wt%, pH=6.5~8.0,40~55 DEG C of temperature, 60~140W of ultrasonic power, the ultrasound works time 3~
5s, 1~3s of intermittent time, 30~60min of extraction time are warming up to 95 DEG C of enzyme deactivation 3min after enzymatic hydrolysis, are originally water-cooled to
Room temperature, filtering, the extracting solution that filtrate is merged with the resulting filtrate of step S2；
S4, it extracting solution is placed in the centrifuge of 6000r/min is centrifuged 10min, bulky grain residue is removed, then with 200
Mesh filter screen filters, and obtains tea extract, and extracting solution progress is high-pressure homogeneous, and homogenization pressure is 18~30MP, homogenisation cycle 5~13
Secondary, the solution after homogeneous is spray-dried, and 175~198 DEG C of inlet air temperature, 85~100 DEG C of leaving air temp to get anti-oxidant function
It can instant tea powder.
Through detecting, water content 3.2wt%, polyphenol content 34.26wt%, peptide content 42.73wt%, 1mg/mL solution
DPPH clearance rate is 20.16%.
Embodiment 2:
A kind of preparation method of anti-oxidation function instant tea powder includes the following steps:
S1, it takes 500g tea powder to be placed in controllable temperature ultrasonic extraction tank, adds 25~32L of distilled water, impregnate 25 minutes；
S2, compound lywallzyme ultrasonic wave added enzymolysis and extraction is added in the soak of step S1, compound lywallzyme is by pectin
Enzyme: cellulase: glusulase=200U:300U:100U is combined, enzyme concentration be tea powder dry weight 0.5wt%~
2.0wt%, pH=6.5~7.5,32~38 DEG C of temperature, 60~115W of ultrasonic power, 3~4s of ultrasound works time, the intermittent time
1~2s, 30~110min of extraction time are filtered after enzymatic hydrolysis, and filtrate and filter residue difference are spare；
S3,20~30L distilled water is added into the filter residue of step S2, adds compound protease ultrasonic wave added enzymatic hydrolysis and mentions
Take, compound protease is by bromelain: alkali protease=200U:300U is combined, and enzyme concentration is tea powder dry weight
1.5wt%~3.0wt%, pH=7.0~8.0,42~58 DEG C of temperature, 70~135W of ultrasonic power, the ultrasound works time 2~
5s, 1~4s of intermittent time, 30~65min of extraction time are warming up to 95 DEG C of enzyme deactivation 3min after enzymatic hydrolysis, are originally water-cooled to
Room temperature, filtering, the extracting solution that filtrate is merged with the resulting filtrate of step S2；
S4, it extracting solution is placed in the centrifuge of 6000r/min is centrifuged 10min, bulky grain residue is removed, then with 200
Mesh filter screen filters, and obtains tea extract, and extracting solution progress is high-pressure homogeneous, and homogenization pressure is 16~28MP, homogenisation cycle 6~14
Secondary, the solution after homogeneous is spray-dried, and 180~200 DEG C of inlet air temperature, 85~99 DEG C of leaving air temp to get anti-oxidant function
It can instant tea powder.
Through detecting, water content 3.5wt%, polyphenol content 36.12wt%, peptide content 40.86wt%, 1mg/mL solution
DPPH clearance rate is 18.42%.
Embodiment 3:
A kind of preparation method of anti-oxidation function instant tea powder includes the following steps:
S1, it takes 500g tea powder to be placed in controllable temperature ultrasonic extraction tank, adds 25~35L of distilled water, impregnate 20 minutes；
S2, compound lywallzyme ultrasonic wave added enzymolysis and extraction is added in the soak of step S1, compound lywallzyme is by pectin
Enzyme: cellulase: glusulase=200U:300U:100U is combined, enzyme concentration be tea powder dry weight 0.8wt%~
2.0wt%, pH=6.2~7.3,33~39 DEG C of temperature, 70~120W of ultrasonic power, 4~5s of ultrasound works time, the intermittent time
2~3s, 40~120min of extraction time are filtered after enzymatic hydrolysis, and filtrate and filter residue difference are spare；
S3,15~30L distilled water is added into the filter residue of step S2, adds compound protease ultrasonic wave added enzymatic hydrolysis and mentions
Take, compound protease is by bromelain: alkali protease=200U:300U is combined, and enzyme concentration is tea powder dry weight
1.0wt%~2.5wt%, pH=6.8~8.0,45~55 DEG C of temperature, 70~140W of ultrasonic power, the ultrasound works time 4~
5s, 3~4s of intermittent time, 30~70min of extraction time are warming up to 95 DEG C of enzyme deactivation 3min after enzymatic hydrolysis, are originally water-cooled to
Room temperature, filtering, the extracting solution that filtrate is merged with the resulting filtrate of step S2；
S4, it extracting solution is placed in the centrifuge of 6000r/min is centrifuged 10min, bulky grain residue is removed, then with 200
Mesh filter screen filters, and obtains tea extract, and extracting solution progress is high-pressure homogeneous, and homogenization pressure is 17~30MP, homogenisation cycle 6~14
Secondary, the solution after homogeneous is spray-dried, and 185~198 DEG C of inlet air temperature, 86~98 DEG C of leaving air temp to get anti-oxidant function
It can instant tea powder.
Through detecting, water content 3.4wt%, polyphenol content 33.46wt%, peptide content 41.53wt%, 1mg/mL solution
DPPH clearance rate is 19.72%.
Embodiment 4:
A kind of preparation method of anti-oxidation function instant tea powder includes the following steps:
S1, it takes 500g tea powder to be placed in controllable temperature ultrasonic extraction tank, adds 23~32L of distilled water, impregnate 28 minutes；
S2, compound lywallzyme ultrasonic wave added enzymolysis and extraction is added in the soak of step S1, compound lywallzyme is by pectin
Enzyme: cellulase: glusulase=200U:300U:100U is combined, enzyme concentration be tea powder dry weight 0.6wt%~
2.4wt%, pH=6.5~7.3,30~38 DEG C of temperature, 80~120W of ultrasonic power, 3~5s of ultrasound works time, the intermittent time
2~3s, 30~100min of extraction time are filtered after enzymatic hydrolysis, and filtrate and filter residue difference are spare；
S3,15~25L distilled water is added into the filter residue of step S2, adds compound protease ultrasonic wave added enzymatic hydrolysis and mentions
Take, compound protease is by bromelain: alkali protease=200U:300U is combined, and enzyme concentration is tea powder dry weight
1.5wt%~2.5wt%, pH=6.8~8.0,40~50 DEG C of temperature, 80~150W of ultrasonic power, the ultrasound works time 1~
3s, 2~4s of intermittent time, 30~75min of extraction time are warming up to 95 DEG C of enzyme deactivation 3min after enzymatic hydrolysis, are originally water-cooled to
Room temperature, filtering, the extracting solution that filtrate is merged with the resulting filtrate of step S2；
S4, it extracting solution is placed in the centrifuge of 6000r/min is centrifuged 10min, bulky grain residue is removed, then with 200
Mesh filter screen filters, and obtains tea extract, and extracting solution progress is high-pressure homogeneous, and homogenization pressure is 16~30MP, homogenisation cycle 6~15
Secondary, the solution after homogeneous is spray-dried, and 170~200 DEG C of inlet air temperature, 87~100 DEG C of leaving air temp to get anti-oxidant function
It can instant tea powder.
Through detecting, water content 3.3wt%, polyphenol content 35.52wt%, peptide content 42.13wt%, 1mg/mL solution
DPPH clearance rate is 21.28%.
Embodiment described above is preferred version of the invention, is not intended to limit the present invention in any form,
There are also other variants and remodeling under the premise of without departing from claim technical solution in one's power.
Claims (10)
1. a kind of preparation method of anti-oxidation function instant tea powder, which is characterized in that described method includes following steps:
S1, it takes tealeaves or/and tea comprehensive processing leftover bits and pieces to smash it through 40~60 meshes, obtains tea powder, tea powder is placed in controllable temperature
In ultrasonic extraction tank, add distilled water immersion 10~30 minutes, the mass ratio of tea powder and distilled water is 1:(10~60)；
S2, compound lywallzyme is added in the soak of step S1, ultrasonic wave added digests, and filters after enzymatic hydrolysis, filtrate and filter
Slag difference is spare；
S3, distilled water and compound protease are added into the filter residue of step S2, ultrasonic wave added digests, and heats up and goes out after enzymatic hydrolysis
Filtrate is merged to obtain extracting solution with the resulting filtrate of step S2 by enzyme, filtering；
S4, by extracting solution centrifugation, filter, after high-pressure homogeneous, spray drying up to anti-oxidation function instant tea powder.
2. the preparation method of anti-oxidation function instant tea powder according to claim 1, which is characterized in that the compound molten wall
Enzyme is that at least three kinds in pectase, cellulase, hemicellulase, driselase and glusulase are compound；The compound protease
For at least two in papain, alkali protease, neutral proteinase, bromelain, flavor protease and mushroom enzyme
It is compound.
3. the preparation method of anti-oxidation function instant tea powder according to claim 2, which is characterized in that the compound molten wall
Enzyme is that pectase, cellulase and glusulase are compound, the pectase, cellulase and glusulase it is compound than for 100~
200U:150~300U:60~100U；The compound protease is that bromelain and alkali protease are compound, the pineapple egg
The compound ratio of white enzyme and alkali protease is 100~200U:180~350U.
4. the preparation method of anti-oxidation function instant tea powder according to claim 1, which is characterized in that compound in step S2
The enzyme concentration of lywallzyme is the 0.5%~3.5% of tea powder weight；The enzyme concentration of compound protease is tea powder weight in step S3
1%~4%.
5. preparation method according to claim 1, which is characterized in that ultrasound used is low powder pulsed ultrasound, step S2
Middle ultrasonic power is 30W~120W, and the ultrasound works time is 3s~5s, and the intermittent time is 1s~3s；Ultrasonic power in step S3
For 50W~150W, the ultrasound works time is 2s~5s, and the intermittent time is 1s~4s.
6. preparation method according to claim 1, which is characterized in that pH is 5.5~8.0 in step S2, and temperature is 20 DEG C
~60 DEG C, enzymolysis time is 10min~120min；PH is 6.5~9.0 in step S3, and temperature is 30 DEG C~70 DEG C, enzymolysis time
For 20min~120min.
7. preparation method according to claim 6, which is characterized in that pH is 6.0~7.5 in step S2, and temperature is 30 DEG C
~50 DEG C, enzymolysis time is 20min~60min；PH is 6.5~8.0 in step S3, and temperature is 40 DEG C~60 DEG C, enzymolysis time
For 30min~80min.
8. preparation method according to claim 1, which is characterized in that in step S3 heating enzyme deactivation temperature be 90 DEG C~
95 DEG C, the enzyme deactivation time is 1min~5min.
9. preparation method according to claim 1, which is characterized in that the pressure of step S4 mesohigh homogeneous be 15MP~
30MP, the number of homogenisation cycle are 5~15 times.
10. preparation method according to claim 1, which is characterized in that the inlet air temperature being spray-dried in step S4 is 160
DEG C~200 DEG C, leaving air temp is 80 DEG C~100 DEG C.
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