CN103773650A - Application of chitosan magnetic bead fixed yeast to brewing blueberry fruit wine - Google Patents
Application of chitosan magnetic bead fixed yeast to brewing blueberry fruit wine Download PDFInfo
- Publication number
- CN103773650A CN103773650A CN201310658608.9A CN201310658608A CN103773650A CN 103773650 A CN103773650 A CN 103773650A CN 201310658608 A CN201310658608 A CN 201310658608A CN 103773650 A CN103773650 A CN 103773650A
- Authority
- CN
- China
- Prior art keywords
- magnetic bead
- yeast
- fruit wine
- blueberry
- chitosan
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Distillation Of Fermentation Liquor, Processing Of Alcohols, Vinegar And Beer (AREA)
Abstract
The invention provides an application of a chitosan magnetic bead fixed yeast to brewing blueberry fruit wine. The invention further provides a brewing method for the blueberry fruit wine. According to the application, a chitosan magnetic bead is used as a carrier and blueberries are used as raw materials; the process is simple and the cost is low; the prepared blueberry fruit wine has good quality and good mouth feel.
Description
Technical field
The invention belongs to brewing fruit wine technical field, the particularly application of chitosan magnetic bead immobilized yeast in blue berry liquor brewing, also relates to a kind of brewing method of blueberry fruit wine.
Background technology
Fruit wine refers to take fruit or wild fruit as raw material, through broken, presses extracting juice, and the class mild drinking wine forming is allocated in the meticulous brews of technical process such as fermentation or immersion.The features such as it is low that fruit wine has alcoholic strength, nutritious, and mouthfeel is good.Fruit wine because of its alcoholic strength lower, so little to the pungency of health, and often drink fruit wine can play refresh oneself, the effect of Ginseng Extract and not being harmful to the health.
The wine that the mankind brewage is the earliest fruit wine, and Sumerian and ancient Egyptian just started to have brewed grape wine before more than 6000 years.This technology can be traced back to before more than 2000 year equally in Chinese history, just had the legend of " ape and monkey wine brewing " in historical book, but that is only to rely on spontaneous fermentation and the fruit wine made.In Tang and Song Dynasty time, brewing grape wine is just commonplace in China, except grape wine, also has numerous kinds such as jujube wine, pear wine, pomegranate wine.Although kind is numerous, all fail in world's wine brewing history, to monopolize one seat as white wine and yellow rice wine.
The history of the artificial making fruit wine of China wants evening a lot, is commonly considered as grape in the time of Han dynasty and just occurs after importing into from the Western Regions.Until Qing Dynasty last years of a dynasty or reign are positioned at the establishment of the Zhang Yu vinify company in Yantai, just indicate the beginning of the large-scale production of China's fruit wine.After founding the state, domestic brewing fruit wine has had development faster, and at present, general varietal wine in the world, at home all can scale operation.
Fruit wine is the low alcohol that utilizes fruit to brewage to form, and because containing abundant nutritive ingredient in fruit, so fruit wine also has abundant nutrition, wherein contains multiple organic acid, carbohydrate, and aromatic alcohol, and in fruit wine, also contain and can strengthen the materials such as the SOD of immunologic function.In fruit wine, also contain very complete amino acid kind simultaneously, contain a large amount of major elements and trace element, the nutritive substances such as VITAMIN, and nutritive ingredients a lot of in the fermenting process of fruit wine are not consumed and are kept in fruit wine.Being exactly these nutritive ingredients of being retained in fruit wine plays an important role to the normal activity that regulates function of human body and maintain human body.Research and propose and drink fruit wine and be conducive to preventing cardiovascular disease, have sterilization, tonic effect, digestion-aid effect.Fruit wine has the function that regulates body metabolism and stimulate circulation, and can prevent the increase of cholesterol, improves cardio and vascular function.Simultaneously anti-ageing in addition, excite effect of liver function and diuresis.In addition the appropriate fruit wine preventing cancer to a certain extent of long-term drinking.
The making method of fruit wine is mainly following flow process: material choice, crushing and beating, the enzyme-added enzyme that goes out, adjusting composition, main secondary fermentation, clarifying treatment, traditional aging process, processing sterilization, finished product fruit wine.
Fruit is after crushing and beating, should add immediately polygalacturonase to improve crushing juice rate, because polygalacturonase can soften and be broken down into pectic acid and galacturonic acid the pectin substance in pulp, improve the content of the soluble solid in slurries, and then raising crushing juice rate, it is clarified more.
Add after polygalacturonase certain hour, crossed leaching juice, in fruit juice, add certain sulphite, make the acid-respons in itself and fruit juice, discharge slowly free SO
2, to play the effect that suppresses miscellaneous bacteria.
While carrying out Primary Fermentation, activated good yeast is linked in fruit juice to be fermented, under certain temperature condition, carries out Primary Fermentation.It should be noted that during the fermentation the bubble lid that fermentation is formed will stir destruction timely, so that fermentation completely.The Primary Fermentation time is generally about six days.
After Primary Fermentation, be secondary fermentation, when secondary fermentation, with the gauze of four layers, it filtered.When filtration, can sneak into certain air, make the yeast recovery of dormancy, then ferment in 20 ℃ of left and right, be generally 10 to 14 days.When secondary fermentation, it should be noted that the air as far as possible making in fermenting container reduces, to prevent from catching acetic bacteria.
After secondary fermentation, obtain blueberry fruit wine finished product through ageing sterilization.Blueberry fruit wine can carry out deepfreeze before finished product, and this is conducive to the increase of maturation and the stability thereof of blueberry fruit wine.Mainly contain 3 effects: the one, improve taste, make unsafe pigment and excessive vitriol separate out and precipitate; The 2nd, the organic substance such as dead yeast, pectin that can accelerate to remain in fruit wine after fermentation accelerates precipitation; The 3rd, can make at low temperatures in blueberry fruit wine and dissolve more oxygen, accelerate the traditional aging process of fruit wine.Deepfreeze is a kind of physical process, and microbic activity may temporarily decline, and can reactivate, thereby deepfreeze can not maintain lasting microbially stable, but clarifying process is had to certain help in the time that temperature is recovered.
In the last few years, in order to improve fruit wine production efficiency, to guarantee safe and efficient production, main fruit wine Cheng Chan state of the world begin one's study novel process, the new technology of brewing fruit wine, mainly comprise the seed selection of fruit wine yeast, the optimization of technique for brewing fruit wine, ageing, clarification, fruit wine such as is combined at the aspect with beer technology.
(1) Breeding Selection of Wine Yeast new technology
1. screen nature yeast.Because the common local flavor of fruit wine of brewageing with the purebred yeast saccharomyces cerevisiae of screening is flat, fruital taste is insufficient, therefore needs some aroma-producing yeasts of seed selection, ester-producing yeast, as Pichia, can produce some aromatoising substances and flavor compound, can improve the local flavor of fruit wine.2. adopt modern breeding means screening fruit wine yeast.By adopting some modern breeding means, as genetic engineering technique, protoplast fusion, selection by mutation and crossbreeding technology etc., make the production traits of bacterial classification that significantly improvement and raising by a relatively large margin occur.
(2) brewing fruit wine novel process
1. Freeze Concentration Technology.Traditional Freeze Concentration Technique of Grape Wine refers to first grape naturally frozen before harvesting, and then the manual grape of plucking after freezing, squeezes the juice, and obtains the higher Sucus Vitis viniferae of the sugared content of minute quantity, accesses yeast and carry out low temperature fermentation in the most backward Sucus Vitis viniferae.For the production of in Freeze Concentration Technology have suspended crystallization method and progressive freeze.In recent years, some new Freeze Concentration Technology are for Food Freeze Concentration, as ice nucleating bacteria (being called for short INA bacterium).Its principle is, INA bacterium can produce the specificity frozen egg white that ice nucleation activity is very strong, this albumen is first synthetic with the very low monomeric form of ice nucleation activity, then in cell, polymerization row becomes the albumen composition that ice nucleation activity is very strong, this albumen composition is as the freezing template of water molecules, the refrigerating process of acceleration water under the bringing out of subzero comparatively high temps, finally obtains high-quality fruit juice concentrate.External existing pertinent literature report, shows that INA bacterium can significantly improve the undercooling point of food, shortens freezing time, thereby saves mass energy; Also can promote larger ice-crystal growth, reduce running cost.2. immobilized yeast technology.At present, immobilization technology is widely used in wine fermentation.
(3) ageing novel process
Ageing is a very important procedure in wine fermentation process, and ageing fully just can make fruit wine quality be guaranteed.In order to improve the production efficiency of fruit wine, shorten fruit wine digestion time, can adopt some physics householder methods, as the physical factors such as optical, electrical, magnetization are urged old, to strengthen the Chen Xiang of fruit wine, also can adopt chemical process as adopted low-level oxidation and adopting catalyzer to accelerate the ageing of fruit wine.
(4) clarification novel process
After wine fermentation finishes, the impurity in clarification fruit wine plays an important role to the quality of fruit wine.Traditional defecation method is to adopt super-cell and back up pad filtering system.At 20th century the mid-80, in Production of Wine, adopt cross-flow technology to replace diatomite coarse filtration and cardboard and circular disc precision filter abroad, cross-flow technology just starts in the use of China.In recent years, Liu Shuhua has found a kind of method that solves precipitation of fruit wine, adopt syrup to carry out lower glue as finings, its principle is, in the change sugar operation of configuration fruit wine, syrup is fully boiled, leave standstill at least 1h, make syrup naturally cooling, temperature is not less than 30 ℃, add in wine, be filtered to clarification.This method does not change former fruit wine technical process, and quality, taste to fruit wine have no effect, fruital taste keep complete, place the several years or-30 ℃ still not muddy below.
(5) fruit wine is combined with beer technology.Beer Shen Shou China consumers in general like, the brewing process of beer and fruit wine is combined, and the fruit beer of producing had both had the mouthfeel that beer is salubrious, have again the special fragrance of fruit wine.The making method of fruit beer mainly contains 3 kinds, and one is to produce fruit beer by fruit juice and wort co-fermentation, and the second is that fruit wine and beer are blent to production fruit beer, and the third is in beer, to add fruit juice or other seasoningss to allocate and form.
Blueberry, English Blueberry by name, is the blue berry meaning.Blueberry is a kind of little berry, and it is blue that fruit is, and color and luster is vivid and wrapped up by one deck white fruit powder, pulp exquisiteness, and seed is minimum.Blueberry sweet and sour taste, and there is pleasant fragrance, can eat raw and also can be made into the use of blueberry goods.Blueberry mainly contains two types, and one is wild low shrub, and fruit tree is short, and fruit particle is less, but the content of anthocyanidin is very high; The second is artificial culture type blueberry, and fruit tree is higher, similar with common fruit tree, and fruit is also larger, and moisture is more, but the content of anthocyanidin is relatively on the low side.In Blueberry, contain very abundant nutritive ingredient, these not only have good health-care effect, also have the functions such as the cranial nerve of preventing is aging, cardiac stimulant, anticancer vessel softening, enhancing human organism immunity.It is reported, whole world blueberry approximately more than 400 is planted, wherein, in state-owned 37 kinds, have wild species to distribute in the area such as the large Xiaoxinanlin Mountains and Hainan in China northeast.
In Blueberry, contain multiple nutritional components, except sugar, acid, V
cin addition, be also rich in multivitamin, as VA, VB, VE etc., anthocyanidin, SOD, protein, arbutin, food fiber, mineral element (K, Fe, Zn, Ca etc.).Analyze according to the blueberry kind to 14 U.S., determine anthocyanidin content in the fresh blueberry of each hectogram up to 163mg, SOD5.39g, protein 500mg, carbohydrate 12.9g, fatty 100mg VA81-100mg, VE2.7-9.5mg, other fruit, as several times even tens times of grape, apples etc.Moreover, Mineral element content in blueberry is also very high, in the fresh blueberry of each hectogram, containing Ca220-920 μ g, Mg114-249 μ g, P98-274 μ g, Fe7.6-30.0 μ g, Zn2.1-4.3 μ g, must reach 2.54% by aminoacids content, more taller than the hawthorn famous to be rich in amino acid.In addition, in blueberry juice, also contain the special composition such as flavones, thereby be chosen as " one of ten large best nutritional food " by U.S.'s Time-Life News.Often edible blueberry goods, vision enhancing significantly, anti-eye strain; Skin nutrition; Delay cranial nerve aging; The microangiopathy being caused by diabetes is had to therapeutic action; Cardiac function enhancing; Prevention senile dementia.
The people such as Svarcova have collected the pertinent data of 1935～2007 years, think that blueberry is rich in the materials such as cyanin pigment and oligomeric proanthocyanidin, flavones, has promoter action to HUMAN HEALTH.For example can be for the rehabilitation of the chronic diseases such as auxiliary cardiovascular disorder, diabetes, cancer.
Pectin content in blueberry is very high, can effectively reduce cholesterol level, prevents arterial blood sclerosis, promotes cardiovascular health; Contained cyanine two pigments have the function of protection to retina, can strengthen eyesight, prevent eyestrain, and improving eyesight is refreshed oneself; Contained vitamin C has cardiac function enhancing, and preventing cancer and cardiopathic effect, can prevent cranial nerve aging, promotes eyesight.The health-care effect of blueberry mainly contains: be powerful antioxidant, improve memory; Preventing cancer; Vision enhancing; Beauty and skin care; Remove objectionable impurities in body; Improve health microcirculation, regulate sleep, reduce the generation of subhealth state.
For blueberry, the shelf time of blueberry fruit wine is longer.And blue berry bartender has stayed the nutrition such as the sugar of contained needed by human body, acid, mineral substance, VITAMIN in blue berry, do not add the additives such as any artificial color, essence, sanitas, the natural green style that keeps Blueberry is the novel fruit wine with nourishing function.
Immobilization technology is with chemistry or physical method, free enzyme or cell to be fixed in the area of space of restriction, both becoming can the catalytic activity of maintenance own, overcome again the weak point of free state enzyme or cell, a kind of biological catalyst that also can reclaim and recycle after successive reaction.
The research of immobilized enzyme starts from the 1950's; nineteen fifty-three, the biologist of Germany made immobilized enzyme for the first time; the later stage sixties 20th century; thousand cigarette one Lang Shouci of Japan in 1969 produce L-amino acid at the applying immobilized L-Aminoacylase of commercial production scale continuously from DL-amino acid, have realized the once change in enzyme application history.After this, the immobilization technology of enzyme develops rapidly, impel enzyme engineering as one independently subject from fermentation engineering, show one's talent.In the academic meeting of international enzyme engineering for the first time of holding for 1971, the unified English name of determining immobilized enzyme is immobilized enzyme, definite expression immobilized enzyme.
The applied research of immobilized enzyme is from immobilization single enzyme carries out a simple step catalyzed reaction, to the two or more enzymes of immobilization simultaneously.At present, much also very comprehensive to the research of enzyme immobilization technology.The research report of the parties concerned is also more and more, and a large amount of summaries and monograph all highlight the achievement of this respect, but immobilized enzyme also exists defect: (1) needs plurality of enzymes synergy just practicable in some reaction process; (2) the difficult separation and purification of some enzyme; (3) some reaction needs the regeneration of coenzyme.Therefore, the practical application of immobilized enzyme is greatly limited, and in this case, people are not only by attention but also transfer on the main source-microorganism of enzyme, so, in 20 century 70 later stages, on the basis of immobilized enzyme, there is immobilized cell technology.Its mechanism is very simple, exactly the microorganism cells highly dense of living is fixed on carrier, under suitable condition, and makes its continuous growth and breeding, forms the biological catalyst of high density, thereby has greatly accelerated speed of response, and throughput is increased substantially.1976, France produced beer and alcohol with fixed yeast cell first, obtains good effect.
Although immobilized cell technology starting is late, its speed that enters production application from conceptual phase has substantially exceeded immobilized enzyme.At present, the immobilized cell of large-scale commercial production application is also many than immobilized enzyme, and it will be more wide can predicting its application prospect.
Compare with free state cell, immobilized cell has following distinguishing feature:
(1) immobilized cell has kept complete structure and the native state of cell, can carry out normal growth and breeding;
(2) immobilized cell has kept the original enzyme of cell system, coenzyme system and metabolic regulation system, can carry out metabolism according to original pathways metabolism, and effectively carry out metabolic regulation;
(3) owing to there being the provide protection of carrier, can improve the plasmid stability of genetic engineering bacterium;
(4) use fixation of microbial cell can be continuously, stably produce, and can separate continuously and stably;
(5) utilize fixation of microbial cell easily to realize automatically and controlling, save labor force.
(6) overcome that biomass cells is too little to be separated more difficultly with the aqueous solution, easily caused the shortcoming of polluting for 2 times, kept production efficiency high, can purifying and keep for a long time the advantage of high-efficiency strain.
The kind of cell is varied, and size is not identical with characteristic yet, therefore conventional microorganism immobilization method also has a lot, in general, roughly can be divided into absorption method, covalent coupling method, crosslinking and the large class of entrapping method four:
1, absorption method: utilize various solid adsorbent, cell is adsorbed on to its surface and makes the fixing method of cell be called absorption method.Conventional solid adsorbent has diatomite, porous ceramics, how empty glass etc.Such as yeast cell can be adsorbed on the surface of the carriers such as many empty potteries and cellular plastics under the condition of pH3-5, makes immobilized cell, for the fermentation of alcohol and beer.While preparing immobilized cell by absorption method, operation is simple, and growth on cell, breeding and metabolism is significantly impact not, but adsorptive power a little less than, carrier adsorption cell is insecure, easily makes it come off, use is subject to certain restrictions.
2, combined techniques: select suitable carrier, make it the process for fixation together with Cell binding by covalent linkage or ionic linkage and be called combined techniques.Adopt the fixing microorganism of covalent coupling method stronger with the bonding force of solid phase carrier, but because reaction is fierce in fixation procedure, operate more complicatedly, thereby to the larger inhibition of microorganism active existence, general applicability is poor.
3, crosslinking: crosslinking and covalent coupling method are similar, makes, between cell, crosslinked action occurs by bifunctional reagent and makes the fixing method of cell be called crosslinking.But the carrier that crosslinking adopts is non-water-soluble, and linking agent adopts glutaraldehyde, bis-diazotized benzidine etc. more.The cell that crosslinking is fixing and carrier connect firmly, and stability is high, but due to crosslinking reaction condition fierceness, have suppressed microbic activity, and the scope of application is narrower.
4, entrapping method: the method for the immobilized cell that cell embedding is made in many empty carriers inside claims entrapping method.Entrapping method is the most frequently used method of cell fixation.Entrapping method can be divided into gel embedding method and semi-permeable membranes entrapping method, be wrapped in the small grid of gel by cell or in the ultra-filtration membrane of semi-permeable membranes polymkeric substance, the method cost is low, simple to operate, less on cytoactive impact, the intensity of the immobilized cell ball of making is higher.But resistance to mass transfer is larger.The carrier of embedding mainly contains calcium alginate gel, carrageenin, gelatin etc.
Entrapping method has good over-all properties, catalytic activity reservation and viability are all higher, and be embedded in applying flexible in reaction engineering (comprising design, the operational stability etc. of reactor), therefore, entrapping method becomes most widely used process for fixation in whole immobilized biocatalyst technology; But the diffusional resistance of entrapping method is larger, the catalytic activity of cell is restricted, be suitable for reacting of small molecules substrate and product.
Since 20 century 70 later stages, the research of immobilized cell is developed rapidly.Fixed cell carrier mainly can be divided into two large classes, and a class is inorganic carrier, as diatomite, how empty glass etc.; Another kind of is organic carrier, can be divided into again two kinds, and one is natural polymer gel carrier, as agar, gelatin and calcium alginate gel etc., and these carrier lifeless matter toxicity, mass transfer performances is good, but intensity is low, under anaerobic easily by biological decomposition.Another kind is organic synthesis high-molecular gel carrier, and as Polyacrylamide (ACRM) gel, polyvinyl alcohol (PVA) gel etc., common intensity is better, but mass-transfer performance, after embedding, cytoactive had to impact.Also occurred in recent years using the natural consumption materials such as the dry and red date of bagasse, glucose to fix cell, these carrier wide material sources, lifeless matter toxicity, can be used in food grade production process.Meanwhile, the application of immobilized cell is very wide, is summed up and is mainly two aspects, is on the one hand to utilize the outer product of the various born of the same parents of fixation of microbial cell fermentative production, as alcohol, amino acid etc.; To utilize immobilized cell to be combined and to make microorganism electrode with various electrodes on the one hand.
Phase early 1980s, Ugelstad has proposed with being with magnetic particulate to carry out isolated cell, after produce and manufacture immunomagnetic beads by the Dynal company of Norway, sell with this, it is named as Dynabead, separable various cells.This immunomagnetic beads because of easy and simple to handle, isolated cell purity is high, retained the features such as the activity of fixed cell is accepted and apply by people.Its appearance has been broken a kind of traditional concept-organism that common people all accept and " ferromagnetic " for no reason at all.Due to the superparamagnetism of magnetic microsphere, so be the breakthrough of revolution for separating bio product.At present, magnetic microsphere, as the carrier of a kind of novel bioseparation technology and targeted drug, gets more and more people's extensive concerning and studies.
Magnetic microsphere refers to that using proper method to make organic polymer be combined formation with inorganic particle has the complex microsphere of certain magnetic and special construction, because magnetic microsphere has numerous characteristics and the magnetic responsiveness of polymer microsphere, not only can by the method such as copolymerization and surface modification give its surface functional group (as-OH ,-COOH ,-CHO ,-NH
2,-SH etc.), be combined with biologically active substances such as other sized molecules or cells, can also utilize magnetic to check the response of foreign field, displacement in magnetic field, utilize this character to position magnetic microsphere, or from surrounding medium, separate very rapidly magnetic microsphere, by magnetic microsphere to the research object application of force.Magnetic microsphere has two kinds of main effects, and carrier function and centrifugation utilize chemistry, physics and Biological Principles, make simple to operateization of many loaded down with trivial details complexity, also can make the cycle of traditional test greatly shorten.Therefore, magnetic microsphere is used widely in many fields such as target administration, cellular segregation, affinity chromatography, nucleic acids research and immobilized enzyme, and magnetic microsphere is also widely used in the fields such as organic and biochemical synthesising biological catalysis and environment/Microbiological detection of foods in addition.
Selecting suitable magneticsubstance is the most important condition of wanting the magnetic microsphere that processability is superior.At us at present in known a large amount of magneticsubstances, only have different materials can be used for making magnetic particle: metal oxide γ-Fe
2o
3, Fe
3o
4, MeFe
2o
4(Me=Co, Mn, Ni), metal Ni, Co, Fe and alloy Fe-Co thereof, Ni-Fe etc.Co(cubic in above-mentioned magneticsubstance) and Fe(cubic) there is higher saturation magnetization, and the saturation magnetization of remaining metal oxide and Ni is lower.Even if there is the Co compared with high saturation and magnetic intensity, the metals such as Fe, but they are easily oxidized to CoO and FeO in air, and these two kinds of oxide compounds are anti-ferromagnetic materials, and this magnetic to metallic substance plays side effect; Secondly, there is toxicity in Ni, Co etc., is subject to strict restriction in application biological, medicine and other fields; Again, it is often very harsh that the magneticsubstance of these pure metal is prepared the processing condition of particulate, and these reasons have all limited the application of the magnetic particle of these high saturation and magnetic intensities to a certain extent.And the oxide compound (Fe of iron
3o
4, γ-Fe
2o
3) because of it has good oxidative stability, lower toxicity (the about 2000mg/kg body weight of LD50, far away higher than clinical application dosage at present), the feature such as be easy to get is often used as magnetic polymer microsphere magnetic particle.
The method of preparing magnetic polymer microsphere is varied, and main method has:
1. in-situ synthesis: be a kind of chemical transformation, be intended to add in magnetic bead preparation process function monomer or microballoon is carried out to rear modification make microballoon with the group that can form with molysite in a large number coordinate bond or ionic linkage.
2. emulsifier-free emulsion polymerization method: do not contain or only contain the letex polymerization of trace (its concentration is less than CMC value) emulsifying agent in reaction process completely, it is the polyreaction new technology growing up on the basis of conventional emulsion polymerization.
3. microemulsion polymerization method: microemulsion polymerization method is prepared a kind of more method that magnetic microsphere is use in recent years, its point Microemulsion Polymerization method and conversed phase micro emulsion copolymerization method.
4. dispersion copolymerization method: refer to that the monomer polymerization generation that is dissolved in organic solvent (or water) by one is insoluble to the polymkeric substance of this solvent, and form the polymerization methods of stable dispersion system.
5. suspension polymerization: cardinal principle is under the condition existing at magnetic particle, suspension stabilizer and tensio-active agent, makes one or more monomers cause homopolymerization or copolymerization on magnetic particle surface with initiator, and magnetic particle is wrapped in to polymkeric substance the inside.
6. Transfer Radical Polymerization: can be used to prepare core-shell type magnetic microsphere.Specific practice is to make it to become macromole evocating agent by modified magnetic inorganic particulate, is then scattered in monomer, obtains core-shell type magnetic microsphere by mass polymerization.
Chitin is the very abundant natural polysaccharides of a kind of reserves, and it is the main component of the shells such as shrimp, crab, silkworm chrysalis and insect, is a kind of very abundant and very useful natural resources.Mierocrystalline cellulose is the abundantest material in the natural organic-compound existing on the earth, and is exactly secondly chitin, and the former is mainly generated by plant, and the latter is mainly generated by animal.Chitin has a lot of significant advantages: protein is had to high-affinity; There are many reactive groups, can be carried out widely chemical modification etc.Because chitin is indissoluble solution under common solvent, make it all more difficult in the processes such as purification, processing, chemical reaction, application is restricted.If but be hydrolyzed under alkaline condition, slough the part ethanoyl in molecule, will be transformed into chitosan.The solubility property of chitosan improves greatly, therefore be also often referred to as soluble chitin, claims again chitin, chitosan, soluble chitin, Viartril-S, chitosan, chitose etc.
Chitosan is unique naturally occurring alkaline polysaccharide, has good adsorptivity, permeability, moulding property, fiberizability, water absorbability and moisture retention.And there is more outstanding bioaffinity, and on chitosan molecule chain, containing abundant hydroxyl and amino, this just makes it be easy to carry out chemically modified and gives several functions.Be positioned at the free hydroxyl on chitosan saccharide residue, can, with multiple organic acid derivative as esterification occurs for acid anhydrides, carboxylic acid halides etc., import aliphatics or the aromatic acyl of different molecular weight, form acylate; Amino generation acylation reaction on saccharide residue can generate acid amides; also can generate western Buddhist alkali with aldehyde generation condensation reaction; can be cross-linked by aldehyde or the acid anhydrides etc. of bifunctional; obtain cancellated insoluble product; crosslinked object is that product is not dissolved; even swelling also very little, character is very stable, and this is used as the carrier of chromatography for them or the carrier of fixing is very important.
The method of preparing chitosan magnetic microsphere is to utilize amino on chitosan and the aldehyde radical of bifunctional to join, and obtains network structure polymer microballoon.The preparation technology of this experiment mainly adopts the preparation method of the suspension crosslinking of E.B.Denkbas: under violent stirring, the aqueous acetic acid of chitosan and magnetic karyomicrosome are scattered in suitable organic medium, under emulsifying agent exists, can form tiny chitosan particle.Under glutaraldehyde effect, chitosan particle can further be cross-linked to form microballoon.Chitosan magnetic microsphere after crosslinked, is insoluble to diluted acid, has higher intensity.
The discriminating of fruit wine is also indispensable link in brewing fruit wine process.Have sense organ to differentiate, chemistry is differentiated two kinds of methods.
Sense organ is differentiated following four kinds of methods.1. fruit wine appearance identification---fruit wine should present Ruby red or santal look, and wine liquid is limpid transparent, has gloss, no suspended substance, precipitation and research of chaotic phenomenon.2. fruit wine fragrance is differentiated---fruit wine generally should have the distinctive fragrance of former fruit, and old wine also should have strong aroma, and is all generally that fruital and aroma are lumped together.Aroma is abundanter, and the quality of wine is better.3. fruit wine flavour is differentiated---should sour and sweet palatability, and mellow pure and free from extraneous odour, sweet type wine is agreeably sweet, and dry type wine will be done and not puckery, must not have outstanding alcohol smell.4. fruit wine wine degree is differentiated---the wine degree of the domestic fruit wine of China is many within the scope of 12～18 degree.
Chemistry discriminating refers to and can use some analytical instrument, as can be with the multiple side such as chromatography or spectrophotometry, measure the carbohydrate in fruit wine, acids, alcoholic strength, vitamins C, flavonoid, tannin, anthocyanidin, aroma component etc. affect the content of the compound of fruit wine quality, evaluate the quality of the quality of fruit wine with this.
Fruit wine, because the nutrition and health care effect that it has enjoys modern to like, becomes the developing direction of current beverage industries.Carry out energetically the R and D of natural fruit wine, development fruit wine is produced, not only can make full use of the fruit resource of China's abundant, improve resource utilization, also can reduce grain consumption, improve alcohol product consumption structure, meet consumer demand, be of value to national health, can effectively promote slight slope and the exploitation of mountain region resource simultaneously.One of heath food that blueberry is approved as the world, has very large development prospect, and blueberry fruit wine is a part wherein.Blue berry drinking utensils has more nourishing function, welcome by everybody.
The immobilization technology of yeast is a kind of new discovery of being not only immobilization technology, is also that the one of wine fermentation new technology embodies simultaneously.In recent years, a lot of to yeast immobilization technology research, but it is little to fix the research of yeast fermentation fruit wine with magnetic bead.
Summary of the invention
Goal of the invention: the object of the present invention is to provide the application of chitosan magnetic bead immobilized yeast in blue berry liquor brewing.
Technical scheme: the invention provides the application of chitosan magnetic bead immobilized yeast in blue berry liquor brewing.
Wherein, described chitosan magnetic bead immobilized yeast adopts following steps to make:
(1) preparation of Z 250 magnetic bead: iron(ic) chloride and ferrous sulfate are dissolved in distilled water, add PEG to mix; Be heated to 50-70 ℃, stir 20-40min reaction, drip aqueous sodium hydroxide solution to solution and turn black completely, adjust pH to 10-12,50-70 ℃ of constant temperature stirs 1-3h; 70-90 ℃ of ageing 20-40min; Ultrasonic 0.5-2h, washs and adjusts pH to neutral, obtains Z 250 magnetic bead;
(2) preparation of chitosan magnetic bead: Z 250 magnetic bead and chitosan are dissolved in acetic acid aqueous solution, stir and pour in dispersion medium simultaneously, add glutaraldehyde to continue to stir, collect with magnet, clean, obtain chitosan magnetic bead;
(3) expansion of yeast is accompanied: yeast is cultivated to 3-5d on yeast culture liquid;
(4) chitosan magnetic bead immobilized yeast: chitosan magnetic bead after swelling 2-3d, is accessed and expands the yeast of accompanying in yeast culture liquid, cultivate 30-40h for 30-35 ℃;
The present invention also provides a kind of brewing method of blueberry fruit wine, comprises the following steps:
(1) preparation of Z 250 magnetic bead: iron(ic) chloride and ferrous sulfate are dissolved in distilled water, add PEG to mix; Be heated to 50-70 ℃, stir 20-40min reaction, drip aqueous sodium hydroxide solution to solution and turn black completely, adjust pH to 10-12,50-70 ℃ of constant temperature stirs 1-3h; 70-90 ℃ of ageing 20-40min; Ultrasonic 0.5-2h, washs and adjusts pH to neutral, obtains Z 250 magnetic bead;
(2) preparation of chitosan magnetic bead: Z 250 magnetic bead and chitosan are dissolved in acetic acid aqueous solution, stir and pour in dispersion medium simultaneously, add glutaraldehyde to continue to stir, collect with magnet, clean, obtain chitosan magnetic bead;
(3) expansion of yeast is accompanied: yeast is cultivated to 3-5d on yeast culture liquid;
(4) chitosan magnetic bead immobilized yeast: chitosan magnetic bead after swelling 2-3d, is accessed and expands the yeast of accompanying in yeast culture liquid, cultivate 30-40h for 30-35 ℃;
(5) preparation of blueberry slurry: by after blueberry making beating, add the pectinase enzymatic hydrolysis 1-3h of the 0.3-0.5% of blueberry total mass, sterilizing 5-10min in the water-bath of 80-90 ℃, sweet water adding adjusts pol to 12-24BX, add calcium carbonate or citric acid and adjust pH to 3-5, obtain blueberry slurry;
(6) Primary Fermentation: the chitosan magnetic bead of immobilized yeast is added in blueberry sauce to room temperature fermentation 5-7d;
(7) secondary fermentation: the fermented liquid of filtration step (6), filtrate adds the chitosan magnetic bead of immobilized yeast, 10-20 ℃ of fermentation 7-14d; Filter.
In step (1), the concentration of aqueous sodium hydroxide solution is 0.5-1.5mol/L; The amount ratio of iron(ic) chloride, ferrous sulfate, distilled water and PEG is 8-10g:10-12g:250-350mL:50-150mL.
In step (2), described dispersion medium comprises that volume ratio is whiteruss, sherwood oil and the tween 80 of 40-60:60-80:1-5.
In step (2), the amount ratio of Z 250 magnetic bead, chitosan, acetic acid aqueous solution, dispersion medium and glutaraldehyde is 1-2g:2-4g:50-150mL:101-145mL:1-3mL.
In step (2), whipping temp is room temperature, and stirring velocity is 500-1500r/min.
The preparation method of yeast culture liquid is: get 200g potato, after stripping and slicing, add water boil, filter, filtrate adds 20g glucose, is settled to 1000mL, sterilizing and get final product.
Beneficial effect: the invention provides the application of chitosan magnetic bead immobilized yeast in blue berry liquor brewing, it is with chitose magnetic bead carrier, and take blueberry as raw material, technique is simple, with low cost, and the blueberry fruit wine quality better, the mouthfeel that make are good.
Accompanying drawing explanation
Fig. 1 is chitosan magnetic bead immobilized yeast micro-imaging figure of the present invention.
Fig. 2 is the influence curve of main fermentation temperature to immobilization fermentation.
Fig. 3 is the influence curve of initial pol to immobilization fermentation.
Fig. 4 is the impact of initial pH value on immobilization fermentation.
Fig. 5 is the change curve of ethanol content with fermentation time.
Fig. 6 is the change curve of pH with fermentation time.
Fig. 7 is V
cwith the change curve of fermentation time.
Fig. 8 is the change curve of reducing sugar with fermentation time.
Fig. 9 is the change curve of anthocyanidin with fermentation time.
Embodiment
Below in conjunction with accompanying drawing, the present invention is made and being further illustrated.
The formula of substratum of the present invention or preparation method:
The preparation method of yeast culture liquid is: take the potato of 200g, be cut into small pieces, add water boil 30min, filter coolingly, add 20g glucose, constant volume is to 1000mL, rear packing sterilizing 20min at 121 ℃.
The evaluation method that the present invention adopts the quality of blue berry wine fermentation comprises sensory evaluation and physics and chemistry discriminating, as follows respectively:
(1) sensory evaluation
Mainly from outward appearance (20 points), smell (40 points), typicalness (10 points), flavour (30 points) four aspect evaluations.
The sensory evaluation index of blueberry fruit wine, in table 1.
Table 1 blueberry fruit wine sensory evaluation standards of grading
[28]
Evaluation method is as follows:
1. triangular flask outside is wiped clean, opened carefully sealed membrane, fruit wine is poured in clean disposable cup, the height of wine in cup is generally 1/4～1/3;
2. under suitable light (not being direct sunlight), cup is lifted to neat eyebrow, observe the color and luster, mixed clearly of blueberry fruit wine;
3. shake gently disposable cup, and repeatedly use nasil perfume (or spice), differentiate fruital, aroma and an unusually sweet smell of fruit wine;
4. light Min mono-end opening fruit wine, should first be distributed in oral cavity equably, then carefully taste;
5. according to the feature of blue berry colours of wine, fragrance, flavour, repeatedly weigh, first respectively fruit wine typicalness, after look for wine body, give a mark and make detailed record according to blueberry fruit wine standards of grading (as table 2-1), finally write general comment or propose improvement idea to this sample.(all good fruit wine must possess: look: have the real color and luster of former fruit; Muddy clearly: limpid, transparent, glossy, without precipitation; Fruital: the peculiar fragrance with former fruit; Aroma: there is mellow aroma, mainly refer to the ageing perfume (or spice) of wine---various ester perfume (or spice); Taste: sour and sweet palatability, graceful mellow, free from extraneous odour.）
(2) physics and chemistry is differentiated
1, pH measures: the pH instrument that label taking is reserved, electrode is taken out from protection liquid, and after repeatedly rinsing with distilled water, paper using blots the globule of electrode, then electrode is entered into detected solution completely, rocks gently the beaker that detected solution is housed.After registration is stable, record data.After electrode is taken out from detected solution, after repeatedly rinsing with distilled water, paper using blots the globule on electrode, carries out the detection of next solution to be measured.
2, V
cassay
[29]: get 2g fermented liquid, add 1% the HAC of 2mL to dissolve after constant volume to 10mL.Get the test tube of 25mL, add 2.0 × 10 of the KI of 0.2mol/L of HAC, the 3mL of the 0.1mol/L of 2.5mL and 2mL
-4the I of mol/L
2.From liquid to be measured, get 1mL and add 25mL test tube, shake up, constant volume, to scale marks, take HAC+KI as reference, is measured in 290nm place, is A, meanwhile, measures HAC+KI+I
2absorbancy be A '.V
ccontent is:
3, the mensuration of reducing sugar: get supernatant liquor 2mL after sample is centrifugal, add the DNS of 1.5mL, shake up, 5min under boiling water bath, takes out cooling rear constant volume to 25mL, adds 1.5mLDNS with 2mL distilled water, is reference, surveys OD at 540nm place.On mark song, find corresponding numerical value.
4, the mensuration of anthocyanidin
[30]: get the sample liquid of 1mL, add 1% hydrochloric acid-ethanol in the ratio of 1:4, lixiviate 1h at 25 ℃.Get above-mentioned sample liquid 1mL, add respectively the HCl-KCl solution of 9mLpH=1.0 and the HAC-NaAC solution of pH=4.5, shake up, then survey OD value at 510nm and 700nm place respectively.Anthocyanidin content is:
A=(A
510-A
700)
pH=1.0-(A
510-A
700)
pH=4.5
5, the mensuration of ethanol content: bio-sensing instrument
6, outward appearance pol
[31]: hand refractometer
The brewing method of blueberry fruit wine, comprises the following steps:
(1) preparation of Z 250 magnetic bead: the NaOH of preparation 1mol/L and 5% PEG solution.Take the ferrous sulfate of 9.75g iron(ic) chloride and 11.94g, be dissolved in the distilled water of 300mL, after mixing, add the PEG of 100mL; Be heated to 60 ℃, constant temperature lower magnetic force stirs 30min, slowly drips the NaOH of 1mol/L until solution becomes black completely.Adjust pH to 11; Under 60 ℃ of constant temperature, stir 2h; Ageing 30min at 80 ℃, ultrasonic 1h; With distilled water and 95% ethanol repetitive scrubbing, then adjust pH to neutral, filter post-drying, mill, obtain Z 250 magnetic bead;
(2) in 5% of the preparation of chitosan magnetic bead: 100mL acetic acid solution, add 1g Z 250 magnetic bead and 2g chitosan, after mixing, solution is to pearl to be poured in the dispersion medium that under room temperature, 1000r/min stirs, wherein, dispersion medium is made up of 50mL whiteruss and 70mL sherwood oil and 2.4mL emulsifying agent Tween-80; After 10min, add 1mL glutaraldehyde, continue to stir; With magnet collection, then use sherwood oil and acetone continuous wash, then remove its peculiar smell with distilled water, until do not have after chemical peculiar smell, in baking oven, 40 ℃ of dry 1d, obtain chitosan magnetic bead;
(3) expansion of yeast is accompanied: yeast is cultivated to 3-5d on yeast culture liquid;
(4) chitosan magnetic bead immobilized yeast: by chitosan magnetic bead swelling 48h of low temperature in yeast culture liquid; Then the yeast spreading cultivation is divided in the triangular flask that will ferment, then puts into swelling good chitosan magnetic bead, then triangular flask is put into shaking table, rotating speed is 110r, cultivates 34h for 34 ℃; By fixing magnetic bead carrier distilled water flushing one time of yeast, then under microscope and fast imaging system, observe, on carrier, whether fix yeast, see Fig. 1;
(5) preparation of blueberry slurry: by after blueberry making beating, add the pectinase enzymatic hydrolysis 1-3h of the 0.3-0.5% of blueberry total mass, sterilizing 5-10min in the water-bath of 80-90 ℃, sweet water adding adjusts pol to 18-24BX, add calcium carbonate or citric acid and adjust pH to 3-5, obtain blueberry slurry;
(6) Primary Fermentation: the chitosan magnetic bead of immobilized yeast is added in blueberry sauce to room temperature fermentation 5-7d;
(7) secondary fermentation: the fermented liquid of filtration step (6), filtrate adds the chitosan magnetic bead of immobilized yeast, 10-20 ℃ of fermentation 7-14d, secondary fermentation can reduce acidity, improve the quality of wine, rear fermentating period seals bottleneck with preservative film, avoids wine liquid by air acidifying or oxidation;
(8) fall bottle: after secondary fermentation finishes, to fall bottle to fruit wine, with long rubber hose by the wine liquid of clarifying in siphonage sucking-off Erlenmeyer flask, reach and clarify the effect of fruit wine;
(9) acid adjustment: because the blueberry moisture in summer is more, pol is low, and acidity is high, so the blue berry vinic acid degree of brewageing is higher, affecting the mouthfeel of fruit wine, must regulate the acidity of blueberry fruit wine, is mainly to add calcium carbonate (1g/L) to adjust pH to 3.5-4 to fruit wine.
(10) fall bottle, clarification: adding of calcium carbonate makes fruit wine unstable, can produce precipitation, again fall bottle, make wine liquid clarification.
(1) preparation of Z 250 magnetic bead: the NaOH of preparation 0.5mol/L and 5% PEG solution.Take the ferrous sulfate of 8g iron(ic) chloride and 10g, be dissolved in the distilled water of 250mL, after mixing, add the PEG of 50mL; Be heated to 50 ℃, constant temperature lower magnetic force stirs 40min, slowly drips the NaOH of 1mol/L until solution becomes black completely; Adjust pH to 10; Under 50 ℃ of constant temperature, stir 3h; Ageing 40min at 70 ℃, ultrasonic 0.5h; With distilled water and 95% ethanol repetitive scrubbing, then adjust pH to neutral, filter post-drying, mill, obtain Z 250 magnetic bead;
(2) in 5% of the preparation of chitosan magnetic bead: 50mL acetic acid solution, add 1.5g Z 250 magnetic bead and 3g chitosan, after mixing, solution is to pearl to be poured in the dispersion medium that under room temperature, 500r/min stirs, wherein, dispersion medium is made up of 40mL whiteruss and 60mL sherwood oil and 1mL emulsifying agent Tween-80; After 10min, add 2mL glutaraldehyde, continue to stir; With magnet collection, then use sherwood oil and acetone continuous wash, then remove its peculiar smell with distilled water, until do not have after chemical peculiar smell, in baking oven, 40 ℃ of dry 1d, obtain chitosan magnetic bead;
(3) expansion of yeast is accompanied: yeast is cultivated to 3d on yeast culture liquid;
(4) chitosan magnetic bead immobilized yeast: by chitosan magnetic bead swelling 48h of low temperature in yeast culture liquid; Then the yeast spreading cultivation is divided in the triangular flask that will ferment, then puts into swelling good chitosan magnetic bead, then triangular flask is put into shaking table, rotating speed is 110r, cultivates 40h for 30 ℃; By fixing magnetic bead carrier distilled water flushing one time of yeast, then under microscope and fast imaging system, observe, on carrier, whether fix yeast, see Fig. 1;
(5) preparation of blueberry slurry: by after blueberry making beating, add 0.3% pectinase enzymatic hydrolysis 3h of blueberry total mass, sterilizing 5min in the water-bath of 80 ℃, sweet water adding adjusts pol to 18-24BX, adds calcium carbonate or citric acid and adjusts pH to 3-5, obtains blueberry slurry;
(6) Primary Fermentation: the chitosan magnetic bead of immobilized yeast is added in blueberry sauce to room temperature fermentation 5d;
(7) secondary fermentation: the fermented liquid of filtration step (6), filtrate adds the chitosan magnetic bead of immobilized yeast, 10 ℃ of fermentation 14d, secondary fermentation can reduce acidity, improve the quality of wine, rear fermentating period seals bottleneck with preservative film, avoids wine liquid by air acidifying or oxidation;
(8) fall bottle: after secondary fermentation finishes, to fall bottle to fruit wine, with long rubber hose by the wine liquid of clarifying in siphonage sucking-off Erlenmeyer flask, reach and clarify the effect of fruit wine;
(9) acid adjustment: because the blueberry moisture in summer is more, pol is low, and acidity is high, so the blue berry vinic acid degree of brewageing is higher, affecting the mouthfeel of fruit wine, must regulate the acidity of blueberry fruit wine, is mainly to add calcium carbonate (1g/L) to adjust pH to 3.5-4 to fruit wine.
(10) fall bottle, clarification: adding of calcium carbonate makes fruit wine unstable, can produce precipitation, again fall bottle, make wine liquid clarification.
(1) preparation of Z 250 magnetic bead: the NaOH of preparation 1.5mol/L and 5% PEG solution.Take the ferrous sulfate of 10g iron(ic) chloride and 12g, be dissolved in the distilled water of 350mL, after mixing, add the PEG of 150mL; Be heated to 70 ℃, constant temperature lower magnetic force stirs 20min, slowly drips the NaOH of 1mol/L until solution becomes black completely; Adjust pH to 12; Under 70 ℃ of constant temperature, stir 1h; Ageing 20min at 90 ℃, ultrasonic 2h; With distilled water and 95% ethanol repetitive scrubbing, then adjust pH to neutral, filter post-drying, mill, obtain Z 250 magnetic bead;
(2) in 5% of the preparation of chitosan magnetic bead: 150mL acetic acid solution, add 2g Z 250 magnetic bead and 4g chitosan, after mixing, solution is to pearl to be poured in the dispersion medium that under room temperature, 1500r/min stirs, wherein, dispersion medium is made up of 60mL whiteruss and 80mL sherwood oil and 5mL emulsifying agent Tween-80; After 10min, add 3mL glutaraldehyde, continue to stir; With magnet collection, then use sherwood oil and acetone continuous wash, then remove its peculiar smell with distilled water, until do not have after chemical peculiar smell, in baking oven, 40 ℃ of dry 1d, obtain chitosan magnetic bead;
(3) expansion of yeast is accompanied: yeast is cultivated to 5d on yeast culture liquid;
(4) chitosan magnetic bead immobilized yeast: by chitosan magnetic bead swelling 72h of low temperature in yeast culture liquid; Then the yeast spreading cultivation is divided in the triangular flask that will ferment, then puts into swelling good chitosan magnetic bead, then triangular flask is put into shaking table, rotating speed is 110r, cultivates 30h for 35 ℃; By fixing magnetic bead carrier distilled water flushing one time of yeast, then under microscope and fast imaging system, observe, on carrier, whether fix yeast, see Fig. 1;
(5) preparation of blueberry slurry: by after blueberry making beating, add 0.5% pectinase enzymatic hydrolysis 1h of blueberry total mass, sterilizing 10min in the water-bath of 90 ℃, sweet water adding adjusts pol to 18-24BX, adds calcium carbonate or citric acid and adjusts pH to 3-5, obtains blueberry slurry;
(6) Primary Fermentation: the chitosan magnetic bead of immobilized yeast is added in blueberry sauce to room temperature fermentation 7d;
(7) secondary fermentation: the fermented liquid of filtration step (6), filtrate adds the chitosan magnetic bead of immobilized yeast, 20 ℃ of fermentation 7d, secondary fermentation can reduce acidity, improve the quality of wine, rear fermentating period seals bottleneck with preservative film, avoids wine liquid by air acidifying or oxidation;
(8) fall bottle: after secondary fermentation finishes, to fall bottle to fruit wine, with long rubber hose by the wine liquid of clarifying in siphonage sucking-off Erlenmeyer flask, reach and clarify the effect of fruit wine;
(9) acid adjustment: because the blueberry moisture in summer is more, pol is low, and acidity is high, so the blue berry vinic acid degree of brewageing is higher, affecting the mouthfeel of fruit wine, must regulate the acidity of blueberry fruit wine, is mainly to add calcium carbonate (1g/L) to adjust pH to 3.5-4 to fruit wine.
(10) fall bottle, clarification: adding of calcium carbonate makes fruit wine unstable, can produce precipitation, again fall bottle, make wine liquid clarification.
(1) impact of main fermentation temperature on blueberry fruit wine quality
Technique and other conditions are identical with embodiment 1, investigate the impact of different main fermentation temperatures (20-32 ℃) on blueberry fruit wine quality, the results are shown in Figure 2.
As shown in Figure 2, leavening temperature is in particular in the activity that affects enzyme on the impact of Immobilized yeast, the solubleness of material and the mobility of plasma membrane.Yeast saccharomyces cerevisiae is that one is had a liking for warm microorganism, and the suitableeest leavening temperature of S. cervisiae is 25 ℃～32 ℃.Temperature, in the time of 20 ℃, has affected the absorption of nutritive substance and the secretion of meta-bolites; In the time of 32 ℃, temperature is relatively high, and the Metabolic activity of yeast can be subject to certain impact, and the fruit wine wine degree of production is low, weak flavor; Under comparatively high temps, pigment is easy to destroyed, and the fruital of blueberry also loses larger; And in the time of 25 ℃, fermentation steadily, is easy to control.
(2) impact of pol on blueberry fruit wine quality
Technique and other conditions are identical with embodiment 1, investigate the impact of different pols (12-24BX) on blueberry fruit wine quality, the results are shown in Figure 3.
As shown in Figure 3, when initial pol is 24BX, the time of fermentation is longer, and residual sugar amount is also higher, the growth that this also can suppress yeast, and the later stage ethanol content that makes to ferment declines, and the sugariness of wine is high, and sharp flavor, bitter taste are heavier, and fruital taste is lighter; In the time that pol drops to 12BX, fermentation stops early, and taste is light, has certain sharp flavor, the inferior quality of wine; In the time that pol is elevated to 20BX, ethanol content is the highest, and just, sharp flavor disappears wine body taste pure.Therefore it is more suitable that pol is controlled at 20BX.
(3) impact of pH on blueberry fruit wine quality
Technique and other conditions are identical with embodiment 1, investigate different pH(3-5) impact on blueberry fruit wine quality, the results are shown in Figure 4.
As shown in Figure 4, in microorganism growth process, overwhelming majority's reaction in body is enzymatic reaction, and enzymatic reaction has an optimum pH scope, and pH affects the absorption of material in the permeability of the stability by affecting membrane structure, cytolemma and the solvability of material.From scheming, pH is that 3.0,3.5,4.0 o'clock ethanol contents are high, and is 4.5,5.0 o'clock, and ethanol content is low.May be that pH is worth increasing the growth that has suppressed yeast, yeast quantity reduces, and is unfavorable for wine fermentation.Best pH value is 4.
Investigate main fermentation temperature, pH, pol to the impact of blueberry fruit wine quality, designed Three factors-levels orthogonal experiment, parameter and the results are shown in Table 2.
As known from Table 2, the factor order that affects blueberry fruit wine ethanol content is: the initial pol of main fermentation temperature >pH>; Affect blueberry fruit wine V
cthe factor order of content is: the initial pol of main fermentation temperature >pH>.Therefore, main fermentation temperature is most important on the impact of blue berry wine fermentation.Realize blueberry fruit wine the most optimum fermentation condition of high ethanol content be A
3b
2c
3, leavening temperature is 32 ℃, and pH is 3.5, and initial pol is 22BX; Realize the highest V of blueberry fruit wine
cthe optimum fermentation condition of content is A
3b
2c
1, leavening temperature is 32 ℃, and pH is 3.5, and initial pol is 18BX.From economic aspect, the optimum fermentation condition of blueberry fruit wine is that leavening temperature is 32 ℃, and pH is 3.5, and initial pol is 18BX.
The variance analysis of orthogonal test, in table 3 and table 4.
The variance analysis of table 3 to ethanol content
Table 4 is to V
cthe analysis of variance table of content
Can be found out ethanol content, V by table 3 and table 4
cthe relation of content and leavening temperature, pH, initial pol is not remarkable.Within the scope of orthogonal test, ethanol content, V
cthe variation of content is not very large, within the scope of acceptable, analyzes from table 3 and table 4, and fermentation parameter is with reference to orthogonal test.
The comparison of embodiment 6 free state yeast fermentations, Immobilized yeast and secondary Immobilized yeast
Select optimal conditions of fermentation respectively with free yeast, fixed yeast, the fermentation of secondary immobilized yeast, as can be seen from Figure 5, when the fermentation initial stage, the rate of change of ethanol content is basic identical, after fermentation 2d, the ethanol content of fixed yeast rises rapidly, and the ethanol content of other fermentations also rises, but it is fast to be not so good as immobilization, and, the final ethanol content of immobilization fermentation is the highest, and secondly, free state yeast ethanol content is minimum for secondary immobilization fermentation.
As can be seen from Figure 6, ferment first three day time, the pH velocity of variation of three experimental group is basic identical, and after three days, velocity of variation starts to occur difference, and since the 4th day, the variation of the pH of three experimental group started to occur gap.Secondary pH fixing and free state fermentation declines very fast, and Ying Jing drops to 2 left and right, and the pH value trend of fixed yeast is steady and in 3.0 left and right, be more conducive to yeast growth.
See Fig. 7, V
cthe important nutrient in fruit, therefore V
ccontent is also to evaluate an important index of fruit wine quality.As can be seen from Figure 7, the V of three experimental group
ccontent all declines in time, and downward trend is that free state is the fastest, and immobilization is the slowest.When fermentation ends, the V in fruit wine
ccontent is in 0.3mg/100mL left and right, and content is lower, and this may be V
cbe easy to oxidation, so when fermentation ends, content is very low.
As can be seen from Figure 8, test declined at first four days all very fast for three groups, after the 4th day, the speed that reducing sugar declines has slowed down on the contrary, this may be the prolongation along with fermentation time, and the environment of yeast growth also can change, and makes it to be not suitable for the growth of yeast, the minimizing of yeast quantity, utilizes sugared ability to decline.When fermentation ends, the reducing sugar content of fixed yeast is the highest, and secondary reducing sugar content fixing and free state yeast is basic identical.
See Fig. 9, in blueberry, anthocyanidin content is very abundant, and anthocyanidin has a lot of health-care effects to human body, so the content that detects anthocyanidin in blueberry fruit wine is an important indicator.As can be seen from Figure 9, the anthocyanidin content of free state yeast experimental group declines the fastest, and the decline of fixed yeast experimental group is the slowest, and more than 40mg/100g, the anthocyanidin content after secondary fixed yeast experimental group fermentation ends and immobilization experimental group approach.
The fruit wine according to the standard of sensory evaluation, the final filtration of three groups of experiments being obtained by art technology expert is given a mark, and calculates average mark and obtains table 5.
The sensory evaluation of table 5 blueberry fruit wine
As can be seen from the table, the fruit wine of immobilization experimental group is all best aspect four, free state experimental group except outward appearance be all the poorest, this may be because, the chitosan of the chitosan magnetic bead appearance once of fermenting likely comes off, the black magnetic bead of the inside parcel is outside exposed, has affected fruit colours of wine.The magnetic bead that ball in the Fig. 1 taking under microscope is wrapped up by chitosan.
Claims (8)
1. the application of chitosan magnetic bead immobilized yeast in blue berry liquor brewing.
2. application as claimed in claim 1, is characterized in that: described chitosan magnetic bead immobilized yeast adopts following steps to make:
(1) preparation of Z 250 magnetic bead: iron(ic) chloride and ferrous sulfate are dissolved in distilled water, add PEG to mix; Be heated to 50-70 ℃, stir 20-40min reaction, drip aqueous sodium hydroxide solution to solution and turn black completely, adjust pH to 10-12,50-70 ℃ of constant temperature stirs 1-3h; 70-90 ℃ of ageing 20-40min; Ultrasonic 0.5-2h, washs and adjusts pH to neutral, obtains Z 250 magnetic bead;
(2) preparation of chitosan magnetic bead: Z 250 magnetic bead and chitosan are dissolved in acetic acid aqueous solution, stir and pour in dispersion medium simultaneously, add glutaraldehyde to continue to stir, collect with magnet, clean, obtain chitosan magnetic bead;
(3) expansion of yeast is accompanied: yeast is cultivated to 3-5d on yeast culture liquid;
(4) chitosan magnetic bead immobilized yeast: chitosan magnetic bead after swelling 2-3d, is accessed and expands the yeast of accompanying in yeast culture liquid, cultivate 30-40h for 30-35 ℃.
3. a brewing method for blueberry fruit wine, is characterized in that: comprise the following steps:
(1) preparation of Z 250 magnetic bead: iron(ic) chloride and ferrous sulfate are dissolved in distilled water, add PEG to mix; Be heated to 50-70 ℃, stir 20-40min reaction, drip aqueous sodium hydroxide solution to solution and turn black completely, adjust pH to 10-12,50-70 ℃ of constant temperature stirs 1-3h; 70-90 ℃ of ageing 20-40min; Ultrasonic 0.5-2h, washs and adjusts pH to neutral, obtains Z 250 magnetic bead;
(2) preparation of chitosan magnetic bead: Z 250 magnetic bead and chitosan are dissolved in acetic acid aqueous solution, stir and pour in dispersion medium simultaneously, add glutaraldehyde to continue to stir, collect with magnet, clean, obtain chitosan magnetic bead;
(3) expansion of yeast is accompanied: yeast is cultivated to 3-5d on yeast culture liquid;
(4) chitosan magnetic bead immobilized yeast: chitosan magnetic bead after swelling 2-3d, is accessed and expands the yeast of accompanying in yeast culture liquid, cultivate 30-40h for 30-35 ℃;
(5) preparation of blueberry slurry: by after blueberry making beating, add the pectinase enzymatic hydrolysis 1-3h of the 0.3-0.5% of blueberry total mass, sterilizing 5-10min in the water-bath of 80-90 ℃, sweet water adding adjusts pol to 12-24BX, add calcium carbonate or citric acid and adjust pH to 3-5, obtain blueberry slurry;
(6) Primary Fermentation: the chitosan magnetic bead of immobilized yeast is added in blueberry sauce to room temperature fermentation 5-7d;
(7) secondary fermentation: the fermented liquid of filtration step (6), filtrate adds the chitosan magnetic bead of immobilized yeast, 10-20 ℃ of fermentation 7-14d; Filter and get final product.
4. the brewing method of a kind of blueberry fruit wine according to claim 3, is characterized in that: in step (1), the concentration of aqueous sodium hydroxide solution is 0.5-1.5mol/L; The amount ratio of iron(ic) chloride, ferrous sulfate, distilled water and PEG is 8-10g:10-12g:250-350mL:50-150mL.
5. the brewing method of a kind of blueberry fruit wine according to claim 3, is characterized in that: in step (2), described dispersion medium comprises that volume ratio is whiteruss, sherwood oil and the tween 80 of 40-60:60-80:1-5.
6. the brewing method of a kind of blueberry fruit wine according to claim 3, it is characterized in that: in step (2), the amount ratio of Z 250 magnetic bead, chitosan, acetic acid aqueous solution, dispersion medium and glutaraldehyde is 1-2g:2-4g:50-150mL:101-145mL:1-3mL.
7. the brewing method of a kind of blueberry fruit wine according to claim 3, is characterized in that: in step (2), whipping temp is room temperature, and stirring velocity is 500-1500r/min.
8. the brewing method of a kind of blueberry fruit wine according to claim 3, is characterized in that: the preparation method of yeast culture liquid is: get 200g potato, after stripping and slicing, add water boil, filter, filtrate adds 20g glucose, is settled to 1000mL, sterilizing and get final product.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310658608.9A CN103773650A (en) | 2014-02-17 | 2014-02-17 | Application of chitosan magnetic bead fixed yeast to brewing blueberry fruit wine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310658608.9A CN103773650A (en) | 2014-02-17 | 2014-02-17 | Application of chitosan magnetic bead fixed yeast to brewing blueberry fruit wine |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103773650A true CN103773650A (en) | 2014-05-07 |
Family
ID=50566417
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310658608.9A Pending CN103773650A (en) | 2014-02-17 | 2014-02-17 | Application of chitosan magnetic bead fixed yeast to brewing blueberry fruit wine |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103773650A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108611222A (en) * | 2018-05-03 | 2018-10-02 | 阜阳市鸿福农业科技股份有限公司 | A kind of preparation method of Chinese medicine antibacterial strawberry wine |
| CN109682669A (en) * | 2018-12-24 | 2019-04-26 | 久为利华（天津）文化发展有限公司 | The detection method of anthocyanidin content in grape wine |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1623656A (en) * | 2004-11-01 | 2005-06-08 | 天津大学 | Photocatalyst of high active nano magnetic compound and its preparation process |
| CN101240075A (en) * | 2008-03-07 | 2008-08-13 | 哈尔滨工业大学 | Method for preparing chitosan magnetic micro-sphere and method for immobilizing yeast by using the magnetic micro-sphere |
| CN102134334A (en) * | 2010-12-29 | 2011-07-27 | 厦门大学 | Method for preparing magnetic chitosan microspheres |
| KR101293715B1 (en) * | 2012-12-27 | 2013-08-08 | 대한민국 | Method for preparing fermented liquor of blueberry |
-
2014
- 2014-02-17 CN CN201310658608.9A patent/CN103773650A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1623656A (en) * | 2004-11-01 | 2005-06-08 | 天津大学 | Photocatalyst of high active nano magnetic compound and its preparation process |
| CN101240075A (en) * | 2008-03-07 | 2008-08-13 | 哈尔滨工业大学 | Method for preparing chitosan magnetic micro-sphere and method for immobilizing yeast by using the magnetic micro-sphere |
| CN102134334A (en) * | 2010-12-29 | 2011-07-27 | 厦门大学 | Method for preparing magnetic chitosan microspheres |
| KR101293715B1 (en) * | 2012-12-27 | 2013-08-08 | 대한민국 | Method for preparing fermented liquor of blueberry |
Non-Patent Citations (2)
| Title |
|---|
| 刘杰，等: "四氧化三铁磁性纳米粒子的制备及表征", 《颗粒学前沿问题研讨会-暨第九界全国颗粒制备与处理研讨会》 * |
| 满都日娃，等: "蓝莓酒酵母固定化载体的研究", 《东北农业大学学报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108611222A (en) * | 2018-05-03 | 2018-10-02 | 阜阳市鸿福农业科技股份有限公司 | A kind of preparation method of Chinese medicine antibacterial strawberry wine |
| CN109682669A (en) * | 2018-12-24 | 2019-04-26 | 久为利华（天津）文化发展有限公司 | The detection method of anthocyanidin content in grape wine |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN1943425B (en) | Fishy smell removing method for spirulina princeps | |
| CN103907987B (en) | A kind of multi-cultur es mixing is continuously fermented and is prepared the method for Haw Apple Vinegar Drink | |
| CN106119055A (en) | A kind of multi-functional ferment vinegar | |
| CN103937637A (en) | Honey wine brewing technology | |
| CN102676333B (en) | Fruit wine production process of citrus fruit | |
| CN109439489A (en) | A kind of preparation method of Pitaya wine | |
| CN101380130A (en) | Rosa roxburghii vinegar beverage and preparation method thereof | |
| CN103103067B (en) | Production method of ice date wine | |
| CN104059836A (en) | Strawberry health wine and brewing method thereof | |
| CN101220331B (en) | Method for producing double-bacterium ferment raspberry full juice fruit wine | |
| CN107475015B (en) | Red-heart pitaya and white-heart pitaya composite fruit wine and preparation method thereof | |
| CN104585827B (en) | A kind of Folium Nelumbinis fermented product and preparation method thereof | |
| CN109077312A (en) | A kind of compound fruit and vegetable fructus arctii ferment and preparation method thereof | |
| CN102899236B (en) | Process method for brewing ginseng vinegar by immobilized fermentation | |
| CN106085799A (en) | A kind of multi-cultur es co-immobilization fermentation Fructus Mori fruit vinegar and preparation method thereof | |
| CN103740529B (en) | A kind of production technique of sweet osmanthus pear fruit wine | |
| CN102845800B (en) | Method for preparing Hami melon vinegar beverage | |
| CN107259260B (en) | Wolfberry fruit vinegar beverage capable of reducing internal heat and promoting digestion and preparation method thereof | |
| CN103602559B (en) | A kind of preparation method of guava fruit wine | |
| CN101270326B (en) | Method for preparing strawberry fermented juice | |
| CN108410633A (en) | A kind of red kiwifruit low wine and preparation method thereof that red yeast rice mixed fermentation is hyperchromic | |
| CN103773650A (en) | Application of chitosan magnetic bead fixed yeast to brewing blueberry fruit wine | |
| CN102352293B (en) | Method for improving banana fruit wine appearance quality stability through membrane filtration | |
| CN104382147A (en) | Herba portulacae and jerusalem artichoke co-fermented beverage and preparation method thereof | |
| CN109593627B (en) | Electric field strengthening brewing process of sea-buckthorn and Chinese wolfberry health-care fruit wine |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20140507 |