CN103013726B - The algae indigo plant green beer of polysaccharide and drinks and beverage and preparation method thereof - Google Patents
The algae indigo plant green beer of polysaccharide and drinks and beverage and preparation method thereof Download PDFInfo
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Abstract
The present invention is a kind of algae indigo plant green beer of polysaccharide and drinks and beverage and preparation method thereof, the algae indigo plant green beer of polysaccharide and drinks and beverage be by extracted first from spirulina holosaccharide, algocyan and extract from the leaf of bamboo chlorophyll, then add in proportion, mixed with edible alcohol and distilled water, algae indigo plant polysaccharide extraction liquid is made by pasteurization sterilization, during this extract solution is added into beer, white wine and series wine product and beverage in proportion, the white wine of green beer and functional form as functional form, modulation fruit wine series wine product and beverage；Wherein, the composition of the algae indigo plant polysaccharide extraction liquid is by weight respectively：Spirulina holosaccharide accounts for 2% 16%, and spirulina algocyan accounts for 0.5% 10%, and bamboo chlorphyl accounts for 1 20%, and edible alcohol accounts for 5 15%, and distilled water accounts for 39 91.5%, and its sum-rate is 100%.The present invention uses the spirulina holosaccharide for eliminating protein, and algocyan is free of protein, is applied in the algae indigo plant green beer of polysaccharide extraction liquid and drinks and beverage, with radiation proof function.
Description
Technical field
The present invention relates to food and drink processing technology industry.More particularly, to addition algae indigo plant polysaccharide extraction liquid beer, its
His drinks and beverage and preparation method thereof.
Background technology
Beer raw material is barley, and in saccharifying, majority is converted into glucose and maltose, only 3.5 degree low wine
Essence, the general character of beer is that (1) has certain nutrition, and (2) mild drinking wine, (3) flavor taste is similar.
It is nutraceutical that beer is recommended in 9th worldwide nutrition food meeting, and the beer annual production of 96 years China is up to 15,000,000
Ton, according to second place of the world.Brewing industry developing direction is to improve beer product quality from now on for China, and squeezing body world beer is first carried out
Row, beer will survive in competition, it is necessary to implement beer science and technology innovation.
How beer develops to quality beer, and its development approach is to improve its trophism and healthcare function, and spirulina is
A kind of ocean green bio containing rich in protein, polysaccharide mineral matter, vitamin and various trace elements etc., for many years
It is applied to medicine, health products and beverage etc..It can not only be added in beer, can also be added in white wine and other tune
In alcoholic drink processed, fruit wine and beverage.
But, the tablet and capsule that Spirulina Products are made with spirulina powder are in the majority, i.e. directly add spirulina powder
It is added in food, causes mouthfeel bad, digestion is not utilized yet and is absorbed, precipitation and suspension is especially produced in the beverage.And
And, the main component extracted by spirulina is generally phycocyanin and polysaccharide, and lacks spirulina chlorophyll content.Meanwhile, spiral
Algae chlorophyll is alkaline matter, can be separated out in acidic beverages and be destroyed nutritional ingredient, it is difficult to dissolved in the beverage, institute
Specially treated must be carried out with order to retain the composition.
At present, the beverage preparation of the especially special spirulina extract of beer is that spirulina chlorophyll, spirulina algae is blue
The composition such as albumen and spirulina polysaccharide is appropriately combined to be obtained being produced dedicated for the spirulina extract of beverage particularly Beer Brewage
Product.The extracting method that the spirulina extract is used mainly includes extracting chlorophyll, the extraction of phycocyanin, combination enzymolysis, heat
The processes such as water extraction, including：(1) extraction of spirulina chlorophyll and its iron receive the preparation of salt；(2) extraction of phycocyanin；(3)
Free amino acid, few skin, the extraction of oligosaccharides：(4) extraction of spirulina polysaccharide.
Above-mentioned spirulina preparation method extracts abundant, gained nutrient solution include spirulina chlorophyll, spirulina phycocyanin,
The compositions such as spirulina polysaccharide, wherein spirulina chlorophyll content reaches more than 8%.Above-mentioned spirulina extract and its preparation side
Method, remains spirulina chlorophyll content in extraction process, and can be used for beverage especially beer addition, and free from environmental pollution,
It is easy to operate, it is easy for industrialized production.
But, the spirulina polysaccharide for extracting at present is generally spirulina Thick many candies, without the flow for making removing protein treatment
Technique, makes its product radiation proof function weaken or without radiation proof function.Its pigment (algocyan, chlorophyll) stability is poor.Deposit
Put the time it is long after can be lost in.The extract solution of above-mentioned preparation, containing polysaccharide and protein high nutrient, is susceptible to go bad,
The holding time is short in storage and transport process and freezing storage, there is rotten risk.
The apparatus and process of existing spirulina extract also has the following disadvantages：
(1) spirulina extract yield is low；(2) there is a little algae fishy smell；(3) easily precipitate；(4) meat can be produced in beer
Eye is difficult the filiform seen, its producing cause is that protein or phycocyanin in spirulina occur with ethyl acetate in beer
The product of reaction；(5) degraded alcohol and malt degree, make beer return life.
Patent No. 00108822.X《Spirulina liquid protects green method》, it is mainly technically characterized by spirulina breeding
General principle is extracted in the outdoor open spirulina culture pond in pond, particularly large area, magnesium and ferro element is added, so that it is green to improve leaf
Element and algocyan content, and reaching the short time, to return spirulina green.The technique is outdoor open culturing pool, and its daylighting is become by weather
Change " leaving it to chance ", light application time and intensity are uncontrollable, influence product quality,
The content of the invention
It is an object of the invention to overcome disadvantages mentioned above and deficiency, a kind of algae indigo plant polysaccharide extraction liquid and its preparation side are proposed
Method, the extract solution is added in beer and other drinks and beverage, is made the algae indigo plant green beer of polysaccharide and drinks and beverage, can be played pre-
Anti- ionising radiation and effect of radioactive particulate.
In order to realize the purpose of the present invention, a kind of algae indigo plant green beer of polysaccharide and drinks and beverage, the algae indigo plant green beer of polysaccharide and
Drinks and beverage be by extracted first from spirulina holosaccharide, algocyan and extract from the leaf of bamboo chlorophyll, then by than
Example addition, and edible alcohol and distilled water are mixed, and algae indigo plant polysaccharide extraction liquid is made by pasteurization sterilization, and this is extracted
Liquid is added in beer, white wine and series wine product and beverage in proportion, the white wine of green beer and functional form as functional form, modulation
Fruit wine series wine product and beverage；
Wherein, the composition of the algae indigo plant polysaccharide extraction liquid is by weight respectively：Spirulina holosaccharide accounts for 2%-16%, spiral shell
Rotation algae algocyan accounts for 0.5%-10%, and bamboo chlorphyl accounts for 1-20%, and edible alcohol accounts for 5-15%, and distilled water accounts for 39-91.5%,
Its sum-rate is 100%.
Algae indigo plant polysaccharide extraction liquid configuration proportion is：Spirulina holosaccharide accounts for 2%-16%, and spirulina algocyan is accounted for
0.5%-5%, bamboo chlorphyl accounts for 1-20%, and edible alcohol accounts for 10%, and distilled water accounts for 49-86.5%, obtains blue liquid and carries
Take liquid；The extract solution and 95-99% the Huang beer of 1-5% are configured to universal green beer；The extract solution and 93- of 3-7%
95% yellow beer is configured to the green beer of health；The extract solution of 4-10% and the yellow beer configuration successful of 90-96% can health cares high
The green beer of type.
Algae indigo plant polysaccharide extraction liquid configuration proportion is：Spirulina holosaccharide accounts for 3-16%, and spirulina algocyan accounts for 0.1-
1%, bamboo chlorphyl accounts for 2-16%, and edible alcohol accounts for 10%, and distilled water accounts for 57-85%, obtains green liquid extract solution；1-
8% extract solution and 92-99% white wine be configured to health algae indigo plant polysaccharide green wine, the extract solution of 3-16% and
84-97% white wine is configured to universal algae indigo plant polysaccharide green wine.
Algae indigo plant polysaccharide extraction liquid configuration proportion is：Spirulina holosaccharide accounts for 3-12%, and spirulina algocyan accounts for 2-
4%, bamboo chlorphyl accounts for 2-4%；The extract solution be blue-green liquid, 1-5% the algae indigo plant the bluish-green extract solution of polysaccharide and
The beverage of 95-99% is configured to fruit wine and beverage series.
The present invention also proposes the preparation method of a kind of algae indigo plant green beer of polysaccharide and drinks and beverage, the algae indigo plant green beer of polysaccharide and
The preparation method of drinks and beverage first by by spirulina holosaccharide, spirulina algocyan and bamboo chlorphyl, adding in proportion,
Mixed with edible alcohol and distilled water, algae indigo plant polysaccharide extraction liquid is made by pasteurization sterilization, then, by this extract solution
It is added in proportion in beer, white wine and series wine product and beverage, the white wine of green beer and functional form as functional form, modulation fruit
Liquor series wine product and beverage；
Wherein, the extraction of the spirulina holosaccharide is first to extract spirulina Thick many candies, and albumen is separated from Thick many candies
Matter, the purified single sterling of acquisition spirulina polysaccharide；Its concrete technology is：Spirulina powder is first carried out into broken wall treatment, with low
Polar solvent removes lipophilic composition, through hot-water extraction or alkali lye cold extraction or uses microwave radiation exaraction, takes supernatant concentration, second
Centrifugation, dry spirulina Thick many candies after the analysis of alcohol alcohol, protein accounts for the 60-70% of dry cell weight in spirulina；
The separation protein, including Sevag methods, foam separating technology, trichloroacetic acid (TCA) method, heating concentration method and
Enzyme process, most preferably carries out polysaccharide extracting process using the two enzymes method of trypsase and papain, measures the protein of residual
Content is 3%；
The purifying is multiple mixing saccharic composition and small points that the Thick many candies obtained by being extracted by Aqueous extracts may contain
Sub- impurity, further grading purification obtains the single sterling of spirulina polysaccharide.
Trypsase that described preparation method is used, that papain two enzymes method carries out polysaccharide extracting process is as follows：Choosing
With the algae powder solution → trypsin hydrolysis → papain hydrolysis → vacuum concentration → alcohol of spirulina powder → be made into 12%
Precipitation → acetone washing → freeze-drying → polysaccharide semifinished product → ion-exchange chromatography → dialysis → gel chromatography → freeze-drying
Vacuum concentration：It is concentrated in vacuo at 70-80 DEG C containing solid content 20%；Alcohol precipitation is separated, 5 times of volumes of addition in concentrate
95% ethanol, makes spirulina polysaccharide be separated out in flocculent deposit, and most of protein and other compositions retain in the solution, 4000r/
Min centrifugations 15min must be precipitated, and obtain 2 polysaccharide components, wherein, the monose composition of component 1 is D-Glucose, D- xyloses, D- half
Lactose and glucuronic acid；The monose of component 2 is constituted：D-Glucose, D-MANNOSE, L- rhamnoses, D- galactolipins, glucose
Aldehydic acid.
, the trypsin acting temperature of the trypsin hydrolysis is 55 DEG C, and trypsin acting pH value is in 6.2-7.2 pancreases
Protease soaking time is 2.5h, and enzyme dosage is 3.0%；The papain carries out the enzyme concentration 3% of secondary hydrolysis, pH value
6.5, operative temperature is 40 DEG C, is incubated 20h.
The ion-exchange chromatography separating polyose carries out separation of polysaccharides using the DEAE-Sep harose posts of 20X1cm：With
The pyridine of 0.01mol/L, PH8.0-hydrochloride buffer ion balance exchange column, the applied sample amount of Thick many candies is 8ml, and flow velocity is
25ml/h, gradient elution is carried out with the pyridine-hydrochloride buffer of 0.01mol/L, PH8.0 and the NaCl of 0-2mol/L, is received respectively
Collection, freeze-drying obtains component 1 and component 2.
The purifying is grading purification, and its method includes：
Ion-exchange cellulose chromatography：Ion exchange and cellulose are chromatographed to combine and are made a series of ion-exchange fibres
Element chromatography, the various polysaccharide of dissolution, for separating carbohydrate；Washed using NaCl solution or phosphate buffer solution gradient in separation process
It is de-, obtain the one-component of different fractions；
The gel filtration is after polysaccharide preparation is separated through DEAE celluloses, further to be purified with gel filtration
To homogeneous polysaccharide sterling, through gel chromatography separating polyose, polysaccharide sample is obtained into monosaccharide component 1 and component 2 through acidolysis, paper chromatography.
The algae indigo plant green beer of polysaccharide first kills it with pasteurization before algae indigo plant polysaccharide extraction liquid is used in mixed way in configuration
Bacterium mixes after fermentation with beer again, in order to avoid producing precipitation, the program of addition can be in beer fermentation, and the brewer's wort after boiling is cold
Preceding to add or added in the lump when brewer's wort is boiled, its flow boils → wine for malt → crushing → saccharification → filtering → brewer's wort
Flower separation → brewer's wort precipitation and cooling.
During the preparation of the algae indigo plant green wine of polysaccharide, the flow that algae indigo plant polysaccharide extraction liquid is added in white wine is：The steaming of raw material
Boil, starch gelatinization → mixed song is closed fermentation, the storage produced alcohol → steaming bucket through saccharification, fermentation or distill axe distillation → base liquor →
Blend → store.
Algae indigo plant polysaccharide extraction liquid of the invention is that chlorophyll is refined from the leaf of bamboo, and the algae indigo plant polysaccharide extraction liquid for being obtained is allocated into
In beer, through lot of experiments detection and pilot scale, with following features：(1) beer is in light green, clear sub-translucent pleasing；(2) former beer
Wine taste mouthfeel is constant；(3) do not precipitated in long-time；(4) do not change former beer alcohol content and do not return life；(5) than former beer
With better nutritivity, drink for a long time with health role；(6) drinking person can be made to improve the sense of drink amount and food, nothing
Any adverse reaction and side effect.Better than at present with the prepared spirulina extract of prior art；
The present invention uses the spirulina holosaccharide for eliminating protein, and algocyan is free of protein, is applied to algae blue many
In the sugared green beer of extract solution and drinks and beverage, there is radiation proof function with this.Radioresistance and radiation proof spirulina polysaccharide must be
Protein rear screw algae holosaccharide is eliminated, it is a kind of powder or crystal of white, and it is by glucose, rhamnose, xylose, gala
Sugar, mannose etc. are constituted.
Brief description of the drawings
Fig. 1 shows the structure of algocyan；
The UV-VIS spectrums of Fig. 2 display algocyans；
Fig. 3 display algocyan IR spectrums.
Specific embodiment
To make the object, technical solutions and advantages of the present invention become more apparent, below in conjunction with specific embodiment, to this hair
Bright further description.
The algae of the invention indigo plant green beer of polysaccharide and drinks and beverage be by extracted from spirulina holosaccharide, algocyan and from
Chlorophyll is extracted in the leaf of bamboo, is then added in proportion and edible alcohol and distilled water mixing and stirring, killed by pasteurization
Bacterium into algae indigo plant polysaccharide extraction liquid.And during this extract solution is added into beer, white wine and series wine product and beverage in proportion, as work(
Series wine product and the beverage such as the green beer of energy type and the white wine of functional form, modulation fruit wine.
First, the composition of algae indigo plant polysaccharide extraction liquid：
Algae indigo plant polysaccharide extraction liquid of the invention extracts chlorophyll by spirulina holosaccharide, spirulina algocyan and from the leaf of bamboo
It is formulated, wherein, its composition accounts for 2%-16% for spirulina holosaccharide, and spirulina algocyan accounts for 0.5%-5%, and leaf of bamboo leaf is green
Element accounts for 1-20%, and edible alcohol accounts for 10%, and distilled water accounts for 49-86.5%, and its sum-rate is 100%.
2nd, algae indigo plant polysaccharide extraction liquid in each composition preparation technology：
1st, the extracting method of spirulina holosaccharide：
1) method that the first prepares spirulina holosaccharide：Spirulina Thick many candies are first extracted, and egg is separated from Thick many candies
White matter, the purified single sterling of acquisition spirulina polysaccharide.
When A, thick extraction spirulina polysaccharide (polysaccharide of Spirulina.PS), first by spirulina powder
Broken wall treatment is carried out, lipophilic composition is removed with low polar solvent, (Microwave-assisted Extraction can be used through hot-water extraction or alkali lye cold extraction
Take), supernatant concentration is taken, centrifugation, dry spirulina Thick many candies after alcohol chromatography.Protein accounts for dry cell weight in spirulina
60~70%, therefore extract spirulina polysaccharide key be effectively to remove isolating protein.
B, separation protein, at present, the method for deproteinized mainly has following several：Sevag methods, foam separating technology, three
Monoxone (TCA) method, heating concentration method, enzyme process.Sevag methods are the classical ways of deproteinized matter, are usually applicable only to except a small amount of
Albumen and it is necessary to repeatedly.TCA methods deproteinized can shorten flow, and the reduction of polysaccharide loss rate is conducive to follow-up purifying.With
Enzymatic degraded Intake Protein mass-energy improves the yield of polysaccharide crude product, but still has more than 30% protein in gained composition.Using double
Enzyme process (trypsase, papain) carries out polysaccharide extracting process, and the protein content for measuring residual is 3%.
C, purifying
Thick many candies obtained by being extracted by Aqueous extracts may must enter one containing multiple mixing saccharic composition and small molecular weight impurities
Step grading purification can just obtain the single sterling of spirulina polysaccharide.It is general to observe that single narrow peak reflects as purity after chromatography
Fixed standard.Conventional purification process has：
(1) ion-exchange cellulose chromatography：Ion exchange and cellulose are chromatographed to combine and are made a series of ion exchanges
Cellulose chromatography is, dissolution various polysaccharide, for separate carbohydrate different to the affinity of different ions based on resin.Separated
Frequently with NaCl solution or phosphate buffer solution gradient elution in journey, the one-component of different fractions can be obtained.Common are
DEAE celluloses and DEAE-Sepharose.
(2) gel filtration：After polysaccharide preparation is separated through DEAE celluloses, need further to purify ability with gel filtration
Obtain homogeneous polysaccharide sterling.Gel filtration is a kind of chromatography side that the gel particle with certain pore size size is holder
Method, can be separated molecule with different size and shape, and the carbohydrate for separating different polymerization degree is especially effective.Sephadex
(Sephadex G), Ago-Gel (Sepharose), polyacrylamide gel (Bio-gel P) is hydrophilic gel, all
It is widely used in the purifying of polysaccharide.The purge process of spirulina polysaccharide typically using Sephadex G100 or SephadexG200,
Sepharyl S100.But Sephadex G are in itself carbohydrates, the sugar eluted on gel is stained with eluent unavoidably,
It is likely to result in the impure of refined polysaccharide.Measurement of the polysaccharide content sulfuric acid anthrone method tracing detection after purification.
2) second more efficient spirulina polysaccharide novel technology for extracting.Use two enzymes method (trypsase, Papain
Enzyme) polysaccharide extracting process is carried out, albumen effect is taken off close to 100%, Thick many candies enter one using ion-exchange chromatography and gel chromatography
Step purifying, and its composition is surveyed by paper chromatography.Thick many candies are purified by DEAE-Sepharose and Sep hadex-G50,2 are obtained
Individual polysaccharide component.The monose composition of component 1 is D-Glucose, D- xyloses, D- galactolipins, glucuronic acid after measured；Component 2
Monose is constituted：D-Glucose, D-MANNOSE, L- rhamnoses, D- galactolipins, glucuronic acid.
Concrete technology is as follows：
The algae powder solution → trypsin hydrolysis → papain hydrolysis → vacuum concentration of spirulina powder → be made into 12%
→ alcohol precipitation → acetone washing → freeze-drying → polysaccharide semifinished product → ion-exchange chromatography → dialysis → gel chromatography → cold
Freeze dry vacuum concentration：It is concentrated in vacuo to containing solid content 20% at 70~80 DEG C.Alcohol precipitation is separated：5 are added in concentrate
95% ethanol of times volume, makes spirulina polysaccharide be separated out in flocculent deposit, and most of protein and other compositions are retained in solution
In, 4000r/min centrifugations 15min must be precipitated.
Trypsin hydrolysis
The determination of trypsase optimum temperature, the optimum temperature of trypsase is 55 DEG C, now digests de- egg
White effect is best.The determination of the most suitable effect pH value of trypsase, trypsase has enzyme higher in the range of 6.2~7.2
Vigor.The determination of trypsin digestion time, protein content is no longer obvious in polysaccharide crude product after trypsin acting 2.5h
Decline, therefore can terminating reaction after albumen enzyme effect 2.5h.The determination of the most suitable actuating quantity of trypsase, enzyme dosage reaches 3.0%
De- albumen effect no longer substantially increases later, so enzyme dosage is defined as 3.0%.
Secondary hydrolysis is carried out using papain
Enzyme concentration 3%, in the buffer solution of PH6.5,40 DEG C, is incubated 20h, often shaking.Enzymolysis liquid is through vacuum concentration, wine
After essence precipitation, it is 3% to measure residual protein content.
Ion-exchange chromatography separating polyose
Separation of polysaccharides is carried out using DEAE-Sep harose posts (20X1cm)：With the pyridine-salt of 0.01mol/L, PH8.0
Acid buffer ion balance exchange column, the applied sample amount of Thick many candies is 8ml, and flow velocity is 25ml/h, with the pyrrole of 0.01mol/L, PH8.0
The NaCl of pyridine-hydrochloride buffer and 0~2mol/L carries out gradient elution, obtains curve shown in Fig. 1.Two peaks of the curve are distinguished
Collect, freeze-drying obtains rough segmentation group 1 and rough segmentation group 2.
Gel chromatography separating polyose
Polysaccharide sample obtains monose composition through acidolysis, paper chromatography：Component 1 is D-Glucose, D- xyloses, D- galactolipins, Portugal
Grape uronic acid；The monose of component 2 is constituted：D-Glucose, D-MANNOSE, L- rhamnoses, D- galactolipins, glucuronic acid.
Raw material contrast 2：《A kind of spirulina extract and its preparation technology》The extraction of spirulina polysaccharide is from after enzymolysis
Centrifugation leaves supernatant in algae-residue, then residue is extracted twice, is centrifuged twice, merges plus neutral proteinase, is obtained after dialysis
Active spirulina polysaccharide solution.Due to being extracted using algae-residue, the spirulina polysaccharide yield for being obtained is little, without radioresistance work(
Energy.
Spirulina polysaccharide (PSP) of the invention is the raw material based on fresh algae mud and algae powder, with the trichloroacetic acid of improvement
Method is extracted, products obtained therefrom polysaccharide mass content up to more than 92%, better than traditional SEVDG methods.Better than above-mentioned residue twice
Centrifugation extraction method.
2nd, in spirulina algocyan extracting method
Algocyan is the pigment extracted from blue-green algae, and it is that nature is to count little bright-coloured natural blue pigment.It is
The tetrapyrrole (see Fig. 1) of open chain, is combined together with protein by thioether bond more.Algocyan solution does not fluoresce,
But combine to form fluorescence salt complex with zinc ion.It is general to be used alone not as blue pigment, it is more mixed with other natural pigments
Close using medium in fruit juice, beverage.Meanwhile, algocyan is the important photosynthetic element for absorbing luminous energy and transmission excitation energy, in light cooperation
There is important meaning in research with originally process.
Phycobniliprotein is extracted from spirulina, the algocyan of its thioether bond that dissociated with methyl alcohol.Its optimal dissociation condition is：Temperature
It is 40 DEG C to spend, and Dissociation time is 24h, V (methyl alcohol): m (phycobniliprotein)=200: 1. recovery rate is up to 1.5%, UV, IR of product
It is consistent with document.Algocyan chromophore is in an acidic solution to compare extended pattern, and in the knot for comparing closure in neutral solution
Structure.
Used material and instrument
UV is determined with Hitachi U-3400 ultraviolet-visual spectrometers；IR Pekin-Elmer983 type determination of infrared spectroscopy；
Silica gel for chromatography is Haiyang Chemical Plant, Qingdao's product；Algae powder is public purchased from the green marine growth health food Limited Liability in Beihai, Guangxi
Department；Other reagents used are pure for analysis.
Preparation method：
1) extraction of phycobniliprotein
After extracting phycobniliprotein by existing method, in 0.1mol/L, pH=7.0 phosphate buffer solutions dialysis 48h, low temperature is dense
Contracting, 95% ethanol precipitation, centrifugation is washed till colourless with petroleum ether, acetone, methyl alcohol successively.Being vacuum dried must be denatured phycobniliprotein.
2) phycobniliprotein methyl alcohol solves algocyan
5g denaturation phycobniliprotein is weighed, adds 1000mli A Chun, 40 DEG C of constant temperature to stir 24h, suction filtration, filtrate is concentrated into
Dry, residue is dissolved in chloroformic solutions of the 8~10ml containing 2.5% methyl alcohol, centrifugation, supernatant in 25 DEG C of rotary evaporated to dryness,
Residue is dissolved in chloroformic solutions of the 2ml containing 2.5% methyl alcohol, 7 times of petroleum ether precipitation algocyans of volume, petroleum ether.
With chloroform/methanol (4/1) to launch system, separated with silica gel thin-layer chromatography, by two indigo plants that Rf values are 0.96 and 0.88
After color colour band is scraped off, methyl alcohol dissolution extraction, vacuum and low temperature rotary evaporated to dryness is used then to be carried with same thin-layer chromatography method again
Single time.Respectively obtain two configurational isomers of algocyan.
Product analysis：
Product is blue powder shape solid, is insoluble in water, is soluble in the organic solvents such as chloroform, benzene, methyl alcohol.To light, heat all
It is unstable, particularly Rf values for 0.88 isomers it is more unstable, be easily changed into purplish red color substance (Rf=0.92).
Rf values are 0.96 ultraviolet such as Fig. 2 of isomers,(pH=3, acid chloroform)；
IR (pellet technique) such as Fig. 3：3308.6cm-1(VN-H), 2923.4CM-1 (V-CH3), 2857.0cm-1
(V-CH2-), 1706.0cm-1,1597.0cm-1(V-CH-CH-)。
Influence of the dissociation temperature to recovery rate
The key that dissociation phycobniliprotein obtains algocyan is dissociation temperature.Because algocyan is to light thermally labile, with dissociation temperature
The rising of degree, the conjugated chain and configuration of algocyan are converted, and gradually become purplish red color substance, thus the recovery rate of algocyan is notable
Reduce.Dissociation temperature and algocyan recovery rate relation such as table 1.
The relation of the temperature of table 1 and algocyan recovery rate
| Temperature/DEG C | 20 | 30 | 40 | 50 | 60 | 65 |
| Recovery rate/% | 0.50 | 1.00 | 1.50 | 1.42 | 0.20 | 0.14 |
Influence of the Dissociation time to recovery rate
It is 40 DEG C to keep dissociation temperature, and Dissociation time is shown in Table 2. as shown in Table 2 with recovery rate relation, with Dissociation time
Extension, the recovery rate of algocyan is improved therewith, and the time to 24h, close to highest, when the time is close to 60h, recovery rate drops again
It is low.This is probably the generation because algocyan flows back in methyl alcohol, is, with balanced reaction [6], to work as reaction with the adduct of its methyl alcohol
Reach when to a certain degree, the composition of reactant mixture no longer changes.But extension over time, algocyan and its methyl alcohol
There is the conversion of conjugated chain and configuration in adduct, and balance is not moved substantially simultaneously, thus algocyan recovery rate again at leisure
Reduce.
The relation of the Dissociation time of table 2 and algocyan recovery rate
| Dissociation time/h | 6 | 12 | 18 | 24 | 36 | 48 | 60 | 72 |
| Recovery rate/% | 0.20 | 0.75 | 1.00 | 1.50 | 1.54 | 1.55 | 1.48 | 1.40 |
Influence of the ratio of methyl alcohol and phycobniliprotein to recovery rate
It is 40 DEG C to keep dissociation temperature, and Dissociation time is 24h, and solid-liquid ratio is V (methyl alcohol)/m (phycobniliprotein) to recovery rate
Relation such as table 3
The relation of the solid-liquid ratio of table 3 and recovery rate
Solid-liquid ratio 50: 1 100: 1 150: 1 200: 1 250: 1 300: 1
Recovery rate/% 1.05 1.38 1.45 1.52 1.58 1.58
As shown in Table 3, influence of the solid-liquid ratio to recovery rate less because algocyan and its methyl alcohol adduct this
In one balanced reaction, methyl alcohol is always excessive, and the ratio for properly increasing methyl alcohol is conducive to eliminating protection of the protein to pigment and makees
With, the content of methyl alcohol adduct is improved, so as to be conducive to balance to algocyan while mobile, recovery rate increases, but reaction
Up to after balance, then the ratio of methyl alcohol is improved, influenceed very small.
Algocyan comformation in solution property analysis
Algocyan belongs to courage trienes compound, and such compound has two major absorbance peaks, one in ultra-violet (UV) band, one
Visual field.According to theoretical calculation and test result indicate that, the ratio between two absworption peak degree QUV visCan be used to speculate their structure
Type changes[7].When molecular configuration is in big volution type, i.e., 4 pyrrole rings compositions one compare the structure of closure, QUV visValue ratio
It is smaller；And when configuration relatively opens, i.e., close to line style, at least one interannular double bond is become by Z-type for 4 arrangements of pyrrole ring
E types, QUV visValue is than larger.In addition, when configuration changes, the displacement untill being accompanied by absorbing.
Two absworption peaks of the algocyan in chloroform, methyl alcohol are shown in Table 4. as shown in Table 4, after adding sour (pH=3), maximum ripple
The peak shift that absorbs long is larger, is because after acid adding, the nitrogen-atoms of C rings forms quarternary ammonium salt, and the position of absorbing wavelength and intensity are all
Vary widely.Red shift is especially big in acid chloroform, it may be possible to after acid adding, and the polarity of chloroform increases, and methyl alcohol is in itself
Polarity it is larger, polarity increases few after acid adding, thus results in maximum wavelength in acid chloroform and absorbs red shift and increases.In chloroform
In, QUV visValue becomes 0.7725, in methyl alcohol, Q by 0.3509UV visValue becomes 0.6081 by 0.3967, it is seen that free chromophore
It is in an acidic solution to compare extended pattern, and in the structure for comparing closure in neutral solution.
The absorption spectrum of the algocyan of table 4
| Solvent | Antu2 I | Antu 2 I | QUV vis |
| Chloroform | A593.3, 0.6524 | A366.1, 1.8590 | 0.3509 |
| Acid chloroform | A643.8, 1.5582 | A367.0, 2.0170 | 0.7725 |
| Methyl alcohol | A665.9, 0.2971 | A363.2, 0.6481 | 0.3967 |
| Acidic methanol | A675.5, 0.3884 | A364.2, 0.6387 | 0.6081 |
3rd, the extracting method of bamboo chlorphyl
The method that the present invention extracts chlorophyll from the leaf of bamboo, the common metal ion of solution and food additives are to leaf of bamboo leaf
The influence of green plain stability and resistance to acids and bases, oxidative resistance, the reducing resistance of the pigment.
Bamboo chlorphyl extraction process of the invention is to soak stabilization chlorophyll using copper sulfate solution in spirulina culture pond,
After leaf of bamboo treatment, algocyan and magnesium and ferro element are added, under artificial light sources irradiation, optical property is caught by algocyan, used
The Luxs of intensity of illumination 5-10 ten thousand, by photosynthesis, make its algocyan catch light and change into efficiently by wavelength 400-700 millimicrons
Energy light quantum, and be transferred in bamboo chlorphyl, cause the high photosynthesis speed of action, make its algocyan and chlorophyll mutual
Conversion, is complexed into blue-green stabilization pigment, so that bamboo chlorphyl stabilization, and improve capacity and output.
Magnesium is the central atom of chlorophyll molecule porphyrin ring, and magnesium deficiency chlorophyll content is reduced, and photosynthesis is obstructed, spirulina
Turn yellow, the formation of iron deficiency influence chlorophyll, either spirulina chlorophyll or bamboo chlorphyl have this general character, using people
Make light source adjustable controllable, bamboo chlorphyl is stablized its production capacity and pigment stability,
Material needed for bamboo chlorphyl extraction process of the present invention is：
It is pure that instrument is chemistry with medicine using medicines such as 722 type spectrophotometers etc. and ethanol, hydrochloric acid.
Extraction process：
Stalk is gone to clean in bamboo, after air-drying.4-6h is soaked with 0.07-0.08% copper-baths, to stablize chlorophyll, then
Rinsed 4-5 times with clear water, air-dried, it is stand-by.After various solvent leaching different times, take the dry leaf of bamboo and shred, with firm dipped leaf of bamboo face water
Bath, filtering, filtrate decompression are distilled to semifluid shape, are poured out and are cooled to room temperature, and survey yield of weighing extracts flow chart as follows：
Adopt the leaf of bamboo-clean-copper sulphate immersion-immersion-formula prepare-addition algocyan and magnesium and ferro element prepare-artificial
Light source irradiates and the Luxs of radiation illumination intensity 5-10 ten thousand, wavelength 400-700 millimicrons, exposure time 1-3 hours-stirring-steam
Evaporate-finished product
Result and analysis：
Effect of the pH value to bamboo chlorphyl
Qualitative results show that bamboo chlorphyl extract solution is soluble in water, and in green, absolute ethyl alcohol extract solution is in blackish green.
Secure ph is 2.0,6.0,8.0,10.0 and 12.0 standard series buffer solution, draw 2ml extract solutions in capacity, Ran Houfen
Do not diluted with standard buffer solution and be settled to 50ml, be that maximum absorbance is surveyed at 413nm in wavelength, be as a result listed in table 1.
Influence of the pH value of table 1 to bamboo chlorphyl
From table 1 it follows that with the increase of pH value, maximum absorbance is more stable at pH >=8, visual observation：
Color is gradually thin out.
Bamboo chlorphyl is in reducing resistance
It is the sodium sulfite solution of diluent preparing series concentration with citric acid solution (0.01M), adds equivalent pigment to carry
Take liquid dilution and be settled to 50mL, maximum absorbance is determined at wavelength 413nm, be as a result listed in table 2
Influence of the sodium sulfite of table 2 to bamboo chlorphyl
From table 2 it can be seen that sodium sulfite has certain colour killing to act on to chlorophyll, but concentration influence is little, this explanation
Bamboo chlorphyl is to weak reductant stabilization.
The oxidative resistance of bamboo chlorphyl
Prepare the H of sharp concentration respectively with 0.01M citric acid solutions2O2Solution, draws 2ml pigment extracts, respectively with matching somebody with somebody
Each solution of system is settled to 50mL, and absorbance A is surveyed at 413, is as a result listed in table 3：
Influence of the hydrogen peroxide of table 3 to bamboo chlorphyl
From table 3 it can be seen that with H2O2Concentration is raised, and the absorbance of solution is in rising trend, finally almost close to 1.Mesh
Result is surveyed, pigment is gradually changed, and after a few minutes, 4~No. 6 colors are all taken off, the oxidative resistance of this explanation bamboo chlorphyl is very
Difference, should be noted that when pigment is processed and is used and avoids and oxidising agent.
Reaction of the sucrose to chlorophyll
The sucrose solution of series concentration is prepared with the citric acid solution of 0.01 μ, 2mL is drawn, pigment extract is in capacity
50mL is settled to, is that 413nm surveys absorbance A in wavelength, be as a result listed in table 4
Influence of the sucrose solution of table 4 to bamboo chlorphyl
From table 4, it can be seen that sucrose solution has a huge impact to bamboo chlorphyl, and make irregular change, table
Bright chlorophyll is very unstable in sucrose.
Effect of the Sodium Benzoate to bamboo chlorphyl
With distillation prepare series concentration PhCOONa solution, draw 2mL pigment extracts in 5mL volumetric flasks, constant volume
Absorbance A is surveyed at 413nm wavelength, table 5 is as a result listed in
Influence of the Sodium Benzoate of table 5 to bamboo chlorphyl
As can be seen from Table 5, influence of the Sodium Benzoate to bamboo chlorphyl is than larger, and irregular change is presented, and this says
Bright chlorophyll is unstable in Sodium Benzoate.
Influential effect of the sodium phosphate to bamboo chlorphyl
With distillation prepare series concentration phosphoric acid solution, draw 2mL pigment extracts in 5mL volumetric flasks, constant volume after
Absorbance A is surveyed at 413nm wavelength, table 6 is as a result listed in
Influence of the sodium phosphate of table 6 to bamboo chlorphyl
As can be seen from Table 6, sodium phosphate makees irregular change to the solution absorbance of bamboo chlorphyl, shows that leaf of bamboo leaf is green
Element is unstable in sodium radio-phosphate,P-32 solution.
Effect of the metal ion to bamboo chlorphyl
With distillation prepare series concentration metal ion solution, draw 2mL pigment extracts in 5mL volumetric flasks, constant volume
Absorbance A is surveyed at 413nm wavelength, table 7 is as a result listed in
Influence of the metal ion of table 7 to bamboo chlorphyl
As can be seen from Table 7, most metal ions have a huge impact to bamboo chlorphyl, and make irregular change
Change.With AL3+And Ca2+The increase of concentration and absorbance gradually rises.
Conclusion
Bamboo chlorphyl is very unstable in acid and alkaline solution, and more stable in pH ＞ 8, thus in pigment processing or
Effect should be carried out in alkaline medium.
Bamboo chlorphyl the effect of resistance to weak oxide be not it is obvious that and the effect of reducing resistance is obvious, so
Should be avoided and oxidising agent in the processing of pigment and when using.
Clearly, part saccharide compound can make absorbance for the influence of organic acid and carbohydrate to bamboo chlorphyl in food
Increase, part organic acid can reduce absorbance.
Influence of the most metal ions to bamboo chlorphyl substantially, and makees irregular change.
Major Nutrient composition is chlorophyll in its extract solution in this patent, and primary raw material extracts spirulina leaf for spirulina
Green element, the present invention is with the leaf of bamboo as raw material：Its price well below spirulina, with low cost excellent；It is public that China possesses 3,800,000
The above of inclining bamboo grove, leaf of bamboo zeroing landing is rotten to be become, and the wasting of resources is surprising, can turn waste into wealth, and huge resource is utilized；Leaf of bamboo leaf is green
Element is applied in the algae indigo plant green beer of polysaccharide and drinks and beverage, makes to the addition of organic germanium etc. in its green beer and drinks and beverage various rich
Eutrophy element.
3rd, the configuration proportion and compound method of different purposes algae indigo plant polysaccharide extraction liquids
1st, algae indigo plant polysaccharide green extract solution configuration proportion is：Spirulina holosaccharide accounts for 2%-16%, and spirulina algocyan is accounted for
0.5%-5%, bamboo chlorphyl accounts for 1-20%, and edible alcohol accounts for 10%, and distilled water accounts for 49-86.5%.
Said extracted liquid is blue liquid, for beer series.Wherein,
" green beer " universal configuration proportion is：Algae indigo plant polysaccharide extraction liquid accounts for 1-5%, and yellow beer accounts for 95-99%.
" green beer " health configuration proportion is：Algae indigo plant polysaccharide extraction liquid accounts for 3-7%, and yellow beer accounts for 93-95%.
" green beer " function health configuration proportion high is：Algae indigo plant polysaccharide extraction liquid accounts for 4-10%, and yellow beer accounts for 90-96%.
The collocation method of the algae indigo plant green beer of polysaccharide：
Because this product is rich in algae indigo plant polysaccharide extraction liquid, first it is mixed with beer again using pasteurization is sterilized before being used in mixed way
After fermentation, in order to avoid producing precipitation, the program of addition can be added or boiling in beer fermentation before the brewer's wort cooling after boiling
Added in the lump during brewer's wort.
Algae indigo plant polysaccharide extraction liquid adds (being configured to universal, health, function health high by above-mentioned three types)
I.e.：Malt → crushing → saccharification → filtering → brewer's wort boils → hops separation → brewer's wort precipitation and cools down
2nd, algae indigo plant polysaccharide green extract solution configuration proportion is：Spirulina holosaccharide accounts for 3-16%, and spirulina algocyan is accounted for
0.1-1%, bamboo chlorphyl accounts for 2-16%, and edible alcohol accounts for 10%, and distilled water accounts for 57-85%.
Said extracted liquid is green liquid, for Liquor series.Wherein,
Universal algae indigo plant polysaccharide green wine configuration proportion：Algae indigo plant polysaccharide green extract solution accounts for 1-8%, and white wine accounts for 92-99%.
Health algae indigo plant polysaccharide green wine configuration proportion：Algae indigo plant polysaccharide green extract solution accounts for 3-16%, and white wine accounts for 84-
97%.
The compound method of the algae indigo plant green wine of polysaccharide：
Algae indigo plant polysaccharide extraction liquid is added in white wine：
The boiling (starch gelatinization) of raw material → mixed song closing fermentation (alcohol is produced in saccharification, fermentation) → steaming bucket or distillation axe distillation
The storage of → base liquor → blend → store → check qualified packaging to dispatch from the factory.
Algae indigo plant polysaccharide green extract solution (being added by above-mentioned white wine modulation ratio)
The compound method of algae indigo plant polysaccharide fruit wine：
(fruit wine self color is yellow, is green after modulation) algae indigo plant polysaccharide blue-green extract solution (is modulated by above-mentioned fruit wine
Ratio is added)
Fruit cleans → removes the peel → crush → squeeze the juice → and secondary filter → chemically examine → ferment → chemically examine → blend → chemically examine →
Ultra filtration → sterilizing → chemical examination → filling → capping → two-stage sterilization → quality inspection → freezing → labeling → pack → dispatch from the factory
3rd, the algae indigo plant bluish-green extract solution configuration proportion of polysaccharide is：Spirulina holosaccharide accounts for 3-12%, and spirulina algocyan accounts for 2-
4%, bamboo chlorphyl accounts for 2-4%.
Said extracted liquid is blue-green liquid, for modulating fruit wine and beverage series.
The compound method of algae indigo plant polysaccharide drink：
" the bluish-green extract solution of algae indigo plant polysaccharide " accounts for 1-5%, and beverage accounts for 95-99%.
Algae of the invention indigo plant polysaccharide extraction liquid, its radiation-resisting functional determines by adding algae blue many sugar amounts, more, its work(of addition
Can be stronger, addition is few, then to weakening, the radiation-resisting functional is ionizing radiation-resistant and radioactive particulate to function phase.And ionize spoke
It refers to industrial electrical equipment, transformer, household appliances (computer, refrigerator, television set, mobile phone etc.) to penetrate, and radioactive particulate refers to nuclear power station
The materials such as the dust on periphery.
Above-mentioned algae indigo plant polysaccharide extraction liquid can be configured to universal, health and Gao Bao in being added to beer and drinks and beverage
Strong type.
It is blue extract solution to be added in beer, and it is green extract solution to be added in white wine, be added to modulation beverage wine and
It is blue-green extract solution in other drinks and beverage.Configuration proportion can be selected according to different drinks true qualities.
Algae indigo plant polysaccharide extraction liquid of the invention with the addition of edible alcohol, be not added with any preservative, can be with the anti-qualitative change of anti-corrosion.
The warehousing storage humidity of algae indigo plant polysaccharide extraction liquid keeps 5-10 DEG C of temperature constant state, moistureproof damp proof, the anti-sun to be exposed to the sun, keep away
Light, 2 years shelf-lifves.If there is 0 DEG C of icing, quality problems will be caused.In transportation, packed using scuttlebutt, prevented
It is inverted, gets rid of and pound, it is damaged.
Green beer, drinks that algae indigo plant polysaccharide extraction liquid of the present invention is made etc., better than prior art Spirulina Green beer, increases
Add the function of radiation proof, fill up the blank of other drinks.
The present invention has increased the function of radiation proof than the Spirulina Green beer that first has, reduces cost, is replaced using bamboo chlorphyl
For spirulina chlorophyll, mountain forest resource, the leaf of bamboo and bamboo grove market are developed, and the organic germanium in the leaf of bamboo is dissolved into leaf of bamboo leaf
In green element, healthcare function is increased.
Algae indigo plant polysaccharide extraction liquid of the invention is by following detection：
1st, report is put a body into a coffin in Jiangxi Province's food quality supervision inspection point inspection；Numbering：980756, per in 640ml/ bottle containing 0.5 gram of algae
Blue polysaccharide extraction liquid, in report after display, product health, physical and chemical index addition extract solution, still reaches China's beer industry mark
It is accurate.
2nd, HSPH of Huaxi Medical Univ survey report：Numbering：990701, per 500ml/ bottles, added in every bottle
0.4 gram of algae indigo plant polysaccharide extraction liquid, has detected part nutritive index.
3rd, Zhongshan University's bio-engineering research center survey report：Detection time 97.6.26, detects that sample is：
A, Luo Xuan Zao Green beers (general nutrition and health care)
B, Luo Xuan Zao Green beers (high-efficient health-care)
C, spirulina algae indigo plant polysaccharide (special efficacy concentrate)
Testing result：Algae indigo plant polysaccharide is active polysaccharide, by glucan, the composition such as rhamnose, there is splendid curative effect to diabetes
And anti-aging effects.
Particular embodiments described above, has been carried out further to the purpose of the present invention, technical scheme and beneficial effect
Describe in detail, should be understood that and the foregoing is only specific embodiment of the invention, be not intended to limit the invention,
All any modification, equivalent substitution and improvements within the spirit and principles in the present invention, done etc., should be included in of the invention
Within protection domain.
Claims (8)
1. the preparation method of a kind of algae indigo plant green beer of polysaccharide, it is characterised in that the preparation method of the algae indigo plant green beer of polysaccharide first by
By spirulina holosaccharide, spirulina algocyan and bamboo chlorphyl, add in proportion, and edible alcohol and distilled water mixing are stirred
Mix, algae indigo plant polysaccharide extraction liquid is made by pasteurization sterilization, then, this extract solution is added in beer in proportion, turn into
The green beer of functional form；
The composition of the algae indigo plant polysaccharide extraction liquid is by weight respectively：Spirulina holosaccharide accounts for 2%-16%, and spirulina algae is blue
Element accounts for 0.5%-10%, and bamboo chlorphyl accounts for 1-20%, and edible alcohol accounts for 5-15%, and distilled water accounts for 39-91.5%, its total amount ratio
Example is 100%；
Wherein, the extraction of the spirulina holosaccharide is first to extract spirulina Thick many candies, and protein is separated from Thick many candies, is passed through
Purifying obtains the single sterling of spirulina polysaccharide；It is described extract spirulina Thick many candies concrete technology be：First spirulina powder is entered
Row broken wall treatment, lipophilic composition is removed with low polar solvent, through hot-water extraction or alkali lye cold extraction or uses microwave radiation exaraction,
Supernatant concentration is taken, centrifugation, dry spirulina Thick many candies after alcohol chromatography；The protein that separated from Thick many candies is utilization
The two enzymes method of trypsase and papain carries out polysaccharide extracting process, and the protein content for measuring residual is 3%；It is described pure
Change is multiple mixing saccharic composition and the small molecular weight impurities that the Thick many candies obtained by being extracted by Aqueous extracts are contained, and is further classified pure
Change obtains the single sterling of spirulina polysaccharide；
The algocyan is the tetrapyrrole of open chain, is combined together by thioether bond with protein more；By existing method
After extracting phycobniliprotein, in 0.1mol/L, pH=7.0 phosphate buffer solutions dialysis 48h, low temperature is concentrated, 95% ethanol precipitation, from
The heart, is washed till colourless with petroleum ether, acetone, methyl alcohol successively；Being vacuum dried must be denatured phycobniliprotein；Weigh denaturation algae courage egg described in 5g
In vain, addition 1000ml methyl alcohol, 40 DEG C of constant temperature, stirring 24h, suction filtration, filtrate is concentrated to dryness, residue is dissolved in 8~10ml and contains
In the chloroformic solution of 2.5% methyl alcohol, centrifugation, in 25 DEG C of rotary evaporated to dryness, residue is dissolved in 2ml containing 2.5% first to supernatant
In the chloroformic solution of alcohol, algocyan described in 7 times of petroleum ether precipitations of volume, petroleum ether obtains the algocyan.
2. a kind of preparation method of the algae indigo plant green beer of polysaccharide according to claim 1, it is characterised in that the spirulina polysaccharide
Preparation method use trypsase, that papain two enzymes method carries out polysaccharide extracting process is as follows：From spirulina powder →
The algae powder solution → trypsin hydrolysis → papain hydrolysis → vacuum concentration → alcohol precipitation → acetone for being made into 12% is washed
Wash → freeze-drying → polysaccharide semifinished product → ion-exchange chromatography → dialysis → gel chromatography → freeze-drying vacuum concentration：
70-80 DEG C is concentrated in vacuo to containing solid content 20%；Alcohol precipitation is separated, and 5 times of 95% ethanol of volume are added in concentrate, is made
Spirulina polysaccharide is separated out in flocculent deposit, and most of protein and other compositions retain in the solution, 4000r/min centrifugations
15min must be precipitated, and obtain 2 polysaccharide components, wherein, the monose of component 1 composition is D-Glucose, D- xyloses, D- galactolipins and
Glucuronic acid；The monose of component 2 is constituted：D-Glucose, D-MANNOSE, L- rhamnoses, D- galactolipins and glucuronic acid.
3. a kind of preparation method of the algae indigo plant green beer of polysaccharide according to claim 1, it is characterised in that the trypsase water
The trypsin acting temperature of solution is 55 DEG C, and trypsin acting pH value is 2.5h, enzyme in 6.2-7.2 trypsase soaking time
Consumption is 3.0%；The papain carries out the enzyme concentration 3% of secondary hydrolysis, pH value 6.5, and operative temperature is 40 DEG C, insulation
20h。
4. a kind of preparation method of the algae indigo plant green beer of polysaccharide according to claim 2, it is characterised in that the ion exchange layer
Analysis separating polyose carries out separation of polysaccharides using the DEAE-Sepharose posts of 20X1cm：With the pyridine-salt of 0.01mol/L, pH8.0
Acid buffer ion balance exchange column, the applied sample amount of Thick many candies is 8ml, and flow velocity is 25ml/h, with the pyrrole of 0.01mol/L, pH8.0
The NaCl of pyridine-hydrochloride buffer and 0-2mol/L carries out gradient elution, collects respectively, and freeze-drying obtains component 1 and component 2.
5. the preparation method of a kind of algae indigo plant polysaccharide white wine, it is characterised in that the preparation method of the algae indigo plant polysaccharide white wine first by
By spirulina holosaccharide, spirulina algocyan and bamboo chlorphyl, add in proportion, and edible alcohol and distilled water mixing are stirred
Mix, algae indigo plant polysaccharide extraction liquid is made by pasteurization sterilization, then, this extract solution is added in white wine in proportion, turn into
The white wine of functional form；
The composition of the algae indigo plant polysaccharide extraction liquid is by weight respectively：Spirulina holosaccharide accounts for 2%-16%, and spirulina algae is blue
Element accounts for 0.5%-10%, and bamboo chlorphyl accounts for 1-20%, and edible alcohol accounts for 5-15%, and distilled water accounts for 39-91.5%, its total amount ratio
Example is 100%；
Wherein, the extraction of the spirulina holosaccharide is first to extract spirulina Thick many candies, and protein is separated from Thick many candies, is passed through
Purifying obtains the single sterling of spirulina polysaccharide；It is described extract spirulina Thick many candies concrete technology be：First spirulina powder is entered
Row broken wall treatment, lipophilic composition is removed with low polar solvent, through hot-water extraction or alkali lye cold extraction or uses microwave radiation exaraction,
Supernatant concentration is taken, centrifugation, dry spirulina Thick many candies after alcohol chromatography；The protein that separated from Thick many candies is utilization
The two enzymes method of trypsase and papain carries out polysaccharide extracting process, and the protein content for measuring residual is 3%；It is described pure
Change is multiple mixing saccharic composition and the small molecular weight impurities that the Thick many candies obtained by being extracted by Aqueous extracts are contained, and is further classified pure
Change obtains the single sterling of spirulina polysaccharide；
The algocyan is the tetrapyrrole of open chain, is combined together by thioether bond with protein more；By existing method
After extracting phycobniliprotein, in 0.1mol/L, pH=7.0 phosphate buffer solutions dialysis 48h, low temperature is concentrated, 95% ethanol precipitation, from
The heart, is washed till colourless with petroleum ether, acetone, methyl alcohol successively；Being vacuum dried must be denatured phycobniliprotein；Weigh denaturation algae courage egg described in 5g
In vain, addition 1000ml methyl alcohol, 40 DEG C of constant temperature, stirring 24h, suction filtration, filtrate is concentrated to dryness, residue is dissolved in 8~10ml and contains
In the chloroformic solution of 2.5% methyl alcohol, centrifugation, in 25 DEG C of rotary evaporated to dryness, residue is dissolved in 2ml containing 2.5% first to supernatant
In the chloroformic solution of alcohol, algocyan described in 7 times of petroleum ether precipitations of volume, petroleum ether obtains the algocyan.
6. a kind of preparation method of algae indigo plant polysaccharide white wine according to claim 5, it is characterised in that the spirulina polysaccharide
Preparation method use trypsase, that papain two enzymes method carries out polysaccharide extracting process is as follows：From spirulina powder →
The algae powder solution → trypsin hydrolysis → papain hydrolysis → vacuum concentration → alcohol precipitation → acetone for being made into 12% is washed
Wash → freeze-drying → polysaccharide semifinished product → ion-exchange chromatography → dialysis → gel chromatography → freeze-drying vacuum concentration：
70-80 DEG C is concentrated in vacuo to containing solid content 20%；Alcohol precipitation is separated, and 5 times of 95% ethanol of volume are added in concentrate, is made
Spirulina polysaccharide is separated out in flocculent deposit, and most of protein and other compositions retain in the solution, 4000r/min centrifugations
15min must be precipitated, and obtain 2 polysaccharide components, wherein, the monose of component 1 composition is D-Glucose, D- xyloses, D- galactolipins and
Glucuronic acid；The monose of component 2 is constituted：D-Glucose, D-MANNOSE, L- rhamnoses, D- galactolipins and glucuronic acid.
7. a kind of preparation method of algae indigo plant polysaccharide white wine according to claim 5, it is characterised in that the trypsase water
The trypsin acting temperature of solution is 55 DEG C, and trypsin acting pH value is 2.5h, enzyme in 6.2-7.2 trypsase soaking time
Consumption is 3.0%；The papain carries out the enzyme concentration 3% of secondary hydrolysis, pH value 6.5, and operative temperature is 40 DEG C, insulation
20h。
8. a kind of preparation method of algae indigo plant polysaccharide white wine according to claim 6, it is characterised in that the ion exchange layer
Analysis separating polyose carries out separation of polysaccharides using the DEAE-Sepharose posts of 20X1cm：With the pyridine-salt of 0.01mol/L, pH8.0
Acid buffer ion balance exchange column, the applied sample amount of Thick many candies is 8ml, and flow velocity is 25ml/h, with the pyrrole of 0.01mol/L, pH8.0
The NaCl of pyridine-hydrochloride buffer and 0-2mol/L carries out gradient elution, collects respectively, and freeze-drying obtains component 1 and component 2.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1035425A (en) * | 1988-12-29 | 1989-09-13 | 江西师范大学 | The manufacture method of blue green alga-spiral alga drink |
| CN1218636A (en) * | 1997-11-27 | 1999-06-09 | 中国科学院水生生物研究所 | Extracting method for nutritive component of spirulina |
| CN102178233A (en) * | 2011-04-22 | 2011-09-14 | 南宁大单生物科技有限责任公司 | Spirulina nutrient solution |
| CN102319255A (en) * | 2011-06-08 | 2012-01-18 | 赵波 | Anti-radiation spriulina polysaccharide organic bio-active iodine and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1035425A (en) * | 1988-12-29 | 1989-09-13 | 江西师范大学 | The manufacture method of blue green alga-spiral alga drink |
| CN1218636A (en) * | 1997-11-27 | 1999-06-09 | 中国科学院水生生物研究所 | Extracting method for nutritive component of spirulina |
| CN102178233A (en) * | 2011-04-22 | 2011-09-14 | 南宁大单生物科技有限责任公司 | Spirulina nutrient solution |
| CN102319255A (en) * | 2011-06-08 | 2012-01-18 | 赵波 | Anti-radiation spriulina polysaccharide organic bio-active iodine and application thereof |
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