CN103406080A - Conjugation linoleic acid microcapsulate with wall material made of bacteria exopolysaccharide through complex coacervation method - Google Patents
Conjugation linoleic acid microcapsulate with wall material made of bacteria exopolysaccharide through complex coacervation method Download PDFInfo
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Abstract
The invention relates to conjugation linoleic acid microcapsulate with a wall material made of bacteria exopolysaccharide through a complex coacervation method. The fermentation liquid of bacteria exopolysaccharide is subject to centrifugation, ethanol settlement, protein removing through chloroform-n-butanol method, dialysis and freeze drying for separation and purification, and lemon polyhexose powder is obtained. Experiments show that the polyhexose has an excellent oxidation prevention effect; according to the invention, gelatin and bacteria exopolysaccharide are used for making composite wall material of the microcapsulate, conjugation linoleic acid is embedded through the complex coacervation method, and the technology of making microcapsulate is studied; the manufacturing method is simple and the effect is good.
Description
Technical field
The invention belongs to the microbial polysaccharide field, be specifically related to the preparation that a kind of microbial polysaccharide with antioxidation activity is the CLA microcapsules of wall material.
Technical background
Polysaccharide all has existence in nature higher plant, algae, bacterium and animal body, distributed pole is wide.Different according to source, polysaccharide is divided into to animal polysaccharide, plant polyose, microbial polysaccharide three classes.Microbial polysaccharide is mainly derived from bacterium and fungi, in the last few years, microbe-derived polysaccharide more and more causes people's extensive attention, especially bacterium and the exocellular polysaccharide of fungi, because it is easy to separation and purification and the output advantages of higher is favored by people quite in industrial production.Polysaccharide of many uses, all have application in fields such as medicine, food, Industrial Wastewater Treatment, cleaning supplies, weaving, papermaking, printing and drilling well, ore dressing, explosive industry.Recent study also finds that the exocellular polysaccharide of various bacteria has very strong BA.Along with the continuous discovery of activated extracellular polysaccharide of bacteria, the research of exocellular polysaccharide also causes people's concern more.Microbial polysaccharide has unique Physiology and biochemistry character, and this provides more wide scope and potential value for polysaccharide in the application of food industry.This is starting point of the present invention just.
Microcapsules technology, refer to utilize natural or synthetic high-polymer encapsulated material (wall material), the small capsule nuclear matter (core) of solid, liquid or even gas is coated and forms diameter a kind of micro-scale technology with semi permeability or sealing cyst membrane of (normally between 5 μ m～400 μ m) in 1 μ m～5000 μ m scopes.Common wall material is all carbohydrate, protide, natural plant gum class etc., due to characteristics such as its emulsibility, film forming, easy drying and low-viscosities, be widely studied, but these materials all can not be obtained good effect as the wall material separately, often adopt several materials mutually to arrange in pairs or groups to improve its embedding effect.Add the microbial polysaccharide with antioxidation activity, can significantly improve the oxidation resistance of microcapsules.
Complex coacervation is to make the wall material with the material of two kinds of opposite charges, the core material emulsion dispersion is in wall material solution, by changing pH, temperature or the concentration of aqueous solution of system, make two kinds of wall materials because the interaction of electric charge makes solubility decline cohesion form microcapsules, prepared microcapsule granule is dispersed in liquid medium by the means such as filtering, centrifugal and collects, pass through again the method dryings such as freeze drying, spray-drying, fluid bed, make free flowable microcapsule granule.This method has advantages of high efficiency and high yield.
Freeze drying is by after thing quick freezing to be dried, then the drying means of under high vacuum condition, ice distillation wherein being removed for steam.Due to the distillation of ice, take away heat and make the whole process of freeze-drying keep the sharp freezing state, be conducive to retain the activity of some biological samples.Therefore with freeze-drying, prepare a kind of interpolation extracellular polysaccharide of bacteria CLA microcapsules.CLA is by the multiple position of the derivative conjugated diene acid of linoleic acid and the general name of geometric isomer, 9c wherein, 11t and 10t, two kinds of isomers of 12c have very strong physiologically active, as a kind of new type functional grease, it has multiple beneficial effects such as strengthening immunity, antiatherosclerosis, anti-oxidant, anticancer, fat reducing.
CLA is the same with other greases, tends to produce a series of lipid peroxide in storage and process.Lipid peroxide is the material that a class contains free radical, unfavorable to health.Preventing and reducing Oxidation of Fat and Oils is an important process that guarantees oil quality stability.Photooxidation, enzymatic oxidation and metal catalytic oxidation are three principal elements that start the lipid autoxidation.It is the wall material that selection has the polysaccharide of antioxidation activity, utilize complex coacervation that CLA is prepared into to microcapsule emulsion, by cryodesiccated mode, microcapsule emulsion is prepared into Powdered again, make CLA and external environment isolated, therefore can reach the effect that delays oxidation, and pressed powder easily stores transportation, have more aborning significant using value.
To sensitivities such as light, heat, metal ions, heat resistance is poor due to the antioxidant in food, when high temperature, very easily decomposes and becomes sour, and has brought difficulty to food processing and preservation.Adopt microcapsule embedded technology can prevent that antioxidant itself is oxidized, improve the bioavailability in body, and be convenient to food processing and preservation, also can reach the purpose of slowly-releasing.Therefore, the present invention be take CLA and is core, and it is the wall material that the extracellular polysaccharide of bacteria of antioxidation activity and gelatin are arranged, and utilizes complex coacervation to prepare the CLA microcapsules.Take stability of emulsion, microcapsules efficiency, microcapsules productive rate, particle shape as investigating index, utilize single factor experiment, determine the optimal parameter for preparing each factor of CLA microcapsules.
Summary of the invention
The object of the invention is to complex coacervation and prepare the CLA microcapsules that extracellular polysaccharide of bacteria is the wall material, the preparation method is simple, respond well, be easy to produce.
The objective of the invention is to be achieved through the following technical solutions:
1, the zymotic fluid of a strain Production by Bacteria exocellular polysaccharide, after the centrifugal 10～20min of 3000～5000rpm/min, get supernatant; Supernatant is after Rotary Evaporators is concentrated, and volume is original 1/4～1/10; Concentrate adds 95% ethanol, and the final concentration that makes polysaccharide is 50～100g/L, stand at low temperature 18～24h, centrifugal acquisition sediment; Sediment, with after deionized water dissolving, adopts chloroform-n-butanol method deproteination, and polysaccharide solution and ratio of solvent are 3/1～5/1, chloroform in solvent: n-butanol is 3/1～5/1; By the polysaccharide solution after de-albumen with flowing water dialysis 18～24h, with deionized water dialysis 18～24h, again by 95% ethanol precipitation, stand at low temperature 18～24h, 3000～5000rpm/min, centrifugal 10～20min obtains sediment, collecting precipitation, through freeze drying 18～24h, obtain faint yellow polysaccharide powder.
The polysaccharide with antioxidation activity that 2, will extract and gelatin are that the wall material prepares the CLA microcapsules by complex coacervation.The CLA of take is core-clad material, and Tween 80, class of department 80 are emulsifying agent, and extracellular polysaccharide of bacteria and gelatin are made microcapsule wall material by complex coacervation, and the mass ratio of core-clad material, wall material, emulsifying agent is 1:1:0.02.By CLA, Tween 80, class of department 80 is added in gelatin solution while stirring, 800rpm/min, under 50 ℃, stir 30～40min and form stable pre-emulsion, then by polysaccharide solution at 500rpm/min, under 50 ℃, dropwise add in pre-emulsion, in the pre-emulsion formed, contain 98～102g/L CLA, 0.5～1g/L Tween 80, 0.5 class of～1.0g/L department 80, 48～52g/L gelatin and 48～52g/L polysaccharide, surplus is water, with 10% acetum, adjusting its pH is 3.2～3.8, reaction 0.5～1.5h, under 500rpm/min, make microcapsule emulsion be cooled to 1～10 ℃, freeze drying obtains exocellular polysaccharide CLA microcapsule product.
3, measure above-mentioned extracellular polysaccharide of bacteria CLA microcapsules antioxidant effect, and compare with gelatin CLA microcapsules, the CLA of same amount, Bearing performance goes out non-oxidizability and is significantly increased.
1) measure extracellular polysaccharide of bacteria CLA microcapsules peroxide values (POV) value, the assay method of POV value is carried out according to GB/T5538-1995 and GB/T5538-2005 standard.
2) measure the scavenging action of extracellular polysaccharide of bacteria CLA microcapsules to hydroxy radical (OH), concrete grammar: get PBS 1.5mL(0.15mol/L, pH7.4), 1, 10-phenanthroline (0.75mmol/L) 1.0mL, freshly prepared copperas solution (0.75mmol/L) 1.0mL, add again variable concentrations extracellular polysaccharide of bacteria CLA microcapsule solution 1.0mL, finally add 0.01% hydrogen peroxide 1.0mL, after mixing, be incubated 30min in 37 ℃ of waters bath with thermostatic control, after cooling, measure light absorption value at the 530nm place, with distilled water, return to zero.Using the gelatin CLA microcapsules, CLA of same concentration as positive control.
Result is calculated as follows:
Clearance rate (%)=(A
Contrast-A
Sample)/(A
Contrast-A
Blank) * 100%
3) measure extracellular polysaccharide of bacteria CLA microcapsules reducing power, concrete grammar: get variable concentrations extracellular polysaccharide of bacteria CLA microcapsules sample liquid 0.5～1.5mL, add PBS 2.0～3.0mL, potassium ferricyanide solution 2.0～3.0mL, mix to be placed in 45～55 ℃ of waters bath with thermostatic control and be incubated 15～25min, add the trichloroacetic acid cessation reaction of 2.0～3.0mL10% after cooling.Shake up rear centrifugally, get supernatant 2.0～3.0mL, add the liquor ferri trichloridi of 2.0～3.0mL distilled water and 0.4～0.6mL0.1%, shake up, standing 5～15min, the distilled water of usining returns to zero as reference solution, measures absorbance under 700nm.Using the gelatin CLA microcapsules, CLA of same concentration as positive control.
4, observe the microscopic pattern of above-mentioned extracellular polysaccharide of bacteria microcapsules.
With the fluorescence inverted microscope, observe extracellular polysaccharide of bacteria CLA microcapsules, result shows particle shape preferably.
The invention provides a kind of new higher extracellular polysaccharide of bacteria of output, treated, obtained purer extracellular polysaccharide of bacteria, it has obvious antioxidation activity.This polysaccharide, through the complex coacervation micro capsule technology, is prepared into the microcapsules of application prospect.Preparation method of the present invention is simple, respond well.
The accompanying drawing explanation
Fig. 1, exocellular polysaccharide extract flow process;
Fig. 2, the preparation of exocellular polysaccharide microcapsules;
The peroxide value (POV) of Fig. 3, extracellular polysaccharide of bacteria CLA microcapsules;
The clearance rate to hydroxy radical (OH) of Fig. 4, extracellular polysaccharide of bacteria CLA microcapsules;
The reducing power of Fig. 5, extracellular polysaccharide of bacteria CLA microcapsules;
The microscopic pattern of Fig. 6, arthrobacterium exocellular polysaccharide CLA microcapsules;
The microscopic pattern of Fig. 7, pseudomonad exocellular polysaccharide CLA microcapsules.
The specific embodiment
Below in conjunction with concrete example, technical scheme of the present invention is described further:
The preparation of embodiment 1, arthrobacterium exocellular polysaccharide microcapsules and the research of non-oxidizability.
The preparation of the first step, arthrobacterium exocellular polysaccharide:
1) the Arthrobacter bacterial strain is preserved from row filter for this laboratory.The seed culture based formulas is as follows: in 1L distilled water, contain glucose (10g), peptone (10g), beef extract (3g), sodium chloride (5g).Fermentative medium formula is as follows: in 1L distilled water, contain sucrose (35g), beef extract (20g), sodium chloride (5g), dipotassium hydrogen phosphate (1g), potassium dihydrogen phosphate (0.5g), manganese sulfate (0.5g), magnesium sulfate (0.5g), calcium chloride (0.5g), culture medium and relevant articles for use are all first sterilizing, inoculate at desinfection chamber.Postvaccinal shaking flask is placed on shaking table, cultivates 40h in 30 ℃, 160r/min.
2) according to 1 described zymotic fluid at the centrifugal 15min of 5000rpm/min, get supernatant; Supernatant through Rotary Evaporators in 45 ℃ concentrated after, volume is original 1/5; In concentrate, add 95% ethanol, the final concentration that makes polysaccharide is 50g/L, and stand at low temperature 24h obtains sediment in the centrifugal 10min of 5000rpm/min; Sediment, with after deionized water dissolving, adopts Sevag method deproteination, and polysaccharide solution and ratio of solvent are 4/1, chloroform in solvent: n-butanol=5/1; By the polysaccharide solution after de-albumen with the flowing water 24h that dialyses, with the deionized water 24h that dialyses, in concentrate, add 95% ethanol, the final concentration that makes polysaccharide is 50g/L, stand at low temperature 18h, the centrifugal 10min collecting precipitation of 5000rpm/min, through freeze drying 18h, obtain faint yellow polysaccharide powder (see figure 1).
Second step, the faint yellow polysaccharide powder of first step gained is made to microcapsule wall material, and is prepared into the CLA microcapsules:
Using gelatin, arthrobacterium exocellular polysaccharide as the wall material, be added to while stirring CLA, Tween 80, class of department 80 in gelatin solution, under 800rpm/min, 50 ℃, stir 30min and form stable pre-emulsion, then under 500rpm/min, 50 ℃, dropwise adding arthrobacterium exocellular polysaccharide solution to pre-emulsion, now in pre-emulsion, contain 50g/L gelatin, 50g/L arthrobacterium exocellular polysaccharide, 100g/L CLA, 1g/L Tween 80, class of 1g/L department 80, surplus is water.Wherein Tween 80, class of department 80 are emulsifying agent.With 10% acetum, adjusting its pH is 3.6, and reaction 1h, make microcapsule emulsion be cooled to 4 ℃ under 500rpm/min, and freeze drying 18h, obtain exocellular polysaccharide CLA microcapsule product (see figure 2).
The 3rd step, to prepare gelatin be wall material CLA microcapsules:
The gelatin of take is the wall material, be added to while stirring CLA, Tween 80, class of department 80 in gelatin solution, under 800rpm/min, 50 ℃, stir 30min and form stable pre-emulsion, then under 500rpm/min, 50 ℃, dropwise adding 20% metabisulfite solution to pre-emulsion, now, in pre-emulsion, contain 50g/L gelatin, 50g/L CLA, 0.5g/L Tween 80, class of 0.5g/L department 80,50g/L sodium sulphate, surplus is water.Under 500rpm/min, make microcapsule emulsion be cooled to 4 ℃, freeze drying 18h, obtain gelatin CLA microcapsule product.
The 4th step, second step and the 3rd step gained microcapsules are carried out to antioxygenic property research:
1) mensuration of arthrobacterium exocellular polysaccharide CLA microcapsules peroxide value (POV) value:
CLA and gelatin CLA microcapsules, arthrobacterium exocellular polysaccharide CLA microcapsules are placed in to 65 ℃ of constant temperature oven oxidations, and sampling at regular intervals detects their peroxide value (POV).With the POV value, mean the oxidation rate of CLA and gelatin CLA microcapsules, arthrobacterium exocellular polysaccharide CLA microcapsules, thereby weigh oxidation resistant effect.The assay method of POV value is carried out according to GB/T5538-1995 and GB/T5538-2005 standard.Finally obtain the antioxidant effect of CLA and gelatin CLA microcapsules, arthrobacterium exocellular polysaccharide CLA microcapsules, the POV value of arthrobacterium exocellular polysaccharide CLA microcapsules is lower than gelatin CLA microcapsules and CLA, and declarative section bacillus exocellular polysaccharide CLA microcapsules oxidation resistance slightly is better than CLA and gelatin CLA microcapsules (see figure 3).
2) scavenging action of arthrobacterium exocellular polysaccharide CLA microcapsules to hydroxy radical (OH)
Get PBS 1.5mL(0.15mol/L, pH7.4), 1,10-phenanthroline (0.75mmol/L) 1mL, freshly prepared copperas solution (0.75mmol/L) 1mL, add again variable concentrations arthrobacterium exocellular polysaccharide CLA microcapsule solution 1mL, finally add 0.01% hydrogen peroxide 1mL, after mixing, be incubated 30min in 37 ℃ of waters bath with thermostatic control, at the 530nm place, measure light absorption value after cooling, return to zero with distilled water.Using the gelatin CLA microcapsules, CLA of same concentration as positive control.Result is calculated as follows (see figure 4):
Clearance rate (%)=(A
Contrast-A
Sample)/(A
Contrast-A
Blank) * 100%
3) mensuration of arthrobacterium exocellular polysaccharide CLA microcapsules reducing power
Get the arthrobacterium exocellular polysaccharide CLA microcapsules sample liquid 1.0mL of variable concentrations, add PBS 2.5mL, potassium ferricyanide solution 2.5mL, mix to be placed in 50 ℃ of waters bath with thermostatic control and be incubated 20min, adds the trichloroacetic acid cessation reaction of 2.5mL10% after cooling.Shake up rear centrifugally, get supernatant 2.5mL, add the liquor ferri trichloridi of 2.5mL distilled water and 0.5mL0.1%, shake up, standing 10min, the distilled water of usining returns to zero as reference solution, measures absorbance under 700nm.Using the gelatin CLA microcapsules, CLA of same concentration as the positive control (see figure 5).
The 5th step, second step gained arthrobacterium exocellular polysaccharide CLA microcapsules are used to the form of microscopic examination microcapsules.
A certain amount of microscapsule powder, by a small amount of water-soluble solution, is drawn to several emulsions on slide, observe under 400 times with the fluorescence inverted microscope, result shows the better (see figure 6) of shape of particle.
The preparation of embodiment 2, pseudomonad exocellular polysaccharide microcapsules and the research of non-oxidizability.
The preparation of the first step, pseudomonad exocellular polysaccharide:
1) pseudomonas strain is preserved from row filter for this laboratory.The seed culture based formulas is as follows: in 1L distilled water, contain glucose (10g), peptone (10g), beef extract (3g), sodium chloride (5g).Fermentative medium formula is as follows: in 1L distilled water, contain sucrose (35g), beef extract (20g), dipotassium hydrogen phosphate (1g), potassium dihydrogen phosphate (0.5g), manganese sulfate (0.5g), magnesium sulfate (1g), sodium chloride (30g).Culture medium and relevant articles for use are all first sterilizing, inoculate at desinfection chamber.Postvaccinal shaking flask is placed on shaking table, cultivates 36h in 30 ℃, 160r/min.
2) according to 1 described zymotic fluid at the centrifugal 15min of 5000rpm/min, get supernatant; Supernatant through Rotary Evaporators in 45 ℃ concentrated after, volume is original 1/5; In concentrate, add 95% long-pending ethanol of triploid, stand at low temperature 24h, making the polysaccharide final concentration is 50g/L, obtains sediment in the centrifugal 10min of 5000rpm/min; Sediment, with after deionized water dissolving, adopts Sevag method deproteination, and polysaccharide solution and ratio of solvent are 4/1, chloroform in solvent: n-butanol=5/1; By the polysaccharide solution after de-albumen with the flowing water 24h that dialyses, with the deionized water 24h that dialyses, then by 95% long-pending ethanol precipitation of triploid, making the polysaccharide final concentration is 50g/L, stand at low temperature 18h, the centrifugal 10min collecting precipitation of 5000rpm/min, through freeze drying 18h, obtain faint yellow polysaccharide powder.(see figure 1).
Second step, the faint yellow polysaccharide powder of first step gained is made to microcapsule wall material, and is prepared into the CLA microcapsules:
Using gelatin, pseudomonad exocellular polysaccharide as the wall material, be added to while stirring CLA, Tween 80, class of department 80 in gelatin solution, under 800rpm/min, 50 ℃, stir 30min and form stable pre-emulsion, then under 500rpm/min, 50 ℃, dropwise adding pseudomonad exocellular polysaccharide solution to pre-emulsion, now in pre-emulsion, contain 50g/L gelatin, 50g/L pseudomonad exocellular polysaccharide, 100/L CLA, 1g/L Tween 80, class of 1g/L department 80, surplus is water.Now with 10% acetum, adjusting its pH is 3.6, and reaction 1h, make microcapsule emulsion be cooled to 4 ℃ under 500rpm/min, and freeze drying 18h, obtain exocellular polysaccharide CLA microcapsule product (see figure 2).
The 3rd step, to prepare gelatin be wall material CLA microcapsules:
The gelatin of take is the wall material, be added to while stirring CLA, Tween 80, class of department 80 in gelatin solution, under 800rpm/min, 50 ℃, stir 30min and form stable pre-emulsion, then under 500rpm/min, 50 ℃, dropwise adding 20% metabisulfite solution to pre-emulsion, now in pre-emulsion, contain 50g/L gelatin, 50g/L CLA, 0.5g/L Tween 80, class of 0.5g/L department 80,50g/L sodium sulphate, surplus is water, now under 500rpm/min, make microcapsule emulsion be cooled to 4 ℃, freeze drying 18h, obtain gelatin CLA microcapsule product.
The 4th step, second step and the 3rd step gained microcapsules are carried out to antioxygenic property research:
1) mensuration of pseudomonad exocellular polysaccharide CLA microcapsules peroxide value (POV) value:
CLA and gelatin CLA microcapsules, pseudomonad exocellular polysaccharide CLA microcapsules are placed in to 65 ℃ of constant temperature oven oxidations, and sampling at regular intervals detects its peroxide value (POV).With the POV value, mean the oxidation rate of CLA and gelatin CLA microcapsules, pseudomonad exocellular polysaccharide CLA microcapsules, thereby weigh oxidation resistant effect.The assay method of POV value is carried out according to GB/T5538-1995 and GB/T5538-2005 standard.Finally obtain the antioxidant effect of CLA and gelatin CLA microcapsules, pseudomonad exocellular polysaccharide CLA microcapsules, the POV value of pseudomonad exocellular polysaccharide CLA microcapsules, lower than gelatin CLA microcapsules and CLA, illustrates that pseudomonad exocellular polysaccharide CLA microcapsules oxidation resistance slightly is better than CLA and gelatin CLA microcapsules (see figure 3).
The peroxide value (POV) of table 1, extracellular polysaccharide of bacteria CLA microcapsules
2) scavenging action of pseudomonad exocellular polysaccharide CLA microcapsules to hydroxy radical (OH):
Get PBS 1.5mL(0.15mol/L, pH7.4), 1,10-phenanthroline (0.75mmol/L) 1mL, freshly prepared copperas solution (0.75mmol/L) 1mL, add again variable concentrations pseudomonad exocellular polysaccharide CLA microcapsule solution 1mL, finally add 0.01% hydrogen peroxide 1mL, after mixing, be incubated 30min in 37 ℃ of waters bath with thermostatic control, at the 530nm place, measure light absorption value after cooling, return to zero with distilled water.Using the gelatin CLA microcapsules, CLA of same concentration as positive control.Result (is shown in
Fig. 4) be calculated as follows:
Clearance rate (%)=(A
Contrast-A
Sample)/(A
Contrast-A
Blank) * 100%
Table 2, the extracellular polysaccharide of bacteria CLA microcapsules clearance rate to hydroxy radical (OH)
3) mensuration of pseudomonad exocellular polysaccharide CLA microcapsules reducing power:
Get the pseudomonad polysaccharide CLA microcapsules sample liquid 1.0mL of variable concentrations, add PBS 2.5mL, potassium ferricyanide solution 2.5mL, mix to be placed in 50 ℃ of waters bath with thermostatic control and be incubated 20min, adds the trichloroacetic acid cessation reaction of 2.5mL10% after cooling.Shake up rear centrifugally, get supernatant 2.5mL, add the liquor ferri trichloridi of 2.5mL distilled water and 0.5mL0.1%, shake up, standing 10min, the distilled water of usining returns to zero as reference solution, measures absorbance under 700nm.Using the gelatin CLA microcapsules, CLA of same concentration as the positive control (see figure 5).
The mensuration of table 3, extracellular polysaccharide of bacteria CLA microcapsules reducing power
The 5th step, second step gained pseudomonad exocellular polysaccharide CLA microcapsules are used to the form of microscopic examination microcapsules:
A certain amount of microscapsule powder, by a small amount of water-soluble solution, is drawn to several emulsions on slide, observe under 400 times with the fluorescence inverted microscope, result shows the better (see figure 7) of shape of particle.
Claims (6)
1. complex coacervation prepares the CLA microcapsules that extracellular polysaccharide of bacteria is the wall material, it is characterized in that concrete operation step is as follows:
The first step, get the zymotic fluid of a strain Production by Bacteria exocellular polysaccharide, after the centrifugal 10～20min of 3000～5000rpm/min, get supernatant; It is original 1/4～1/10 that supernatant is concentrated into volume through Rotary Evaporators; In concentrate, add 95% ethanol, the final concentration that makes polysaccharide is 50～100g/L, stand at low temperature 18～24h, and the centrifugal 10～20min of 3000～5000rpm/min obtains sediment;
Second step, by the sediment of first step gained with after deionized water dissolving, adopt chloroform-n-butanol method deproteination, polysaccharide solution is 3/1～5/1 with the solvent volume ratio, chloroform in solvent: the volume ratio of n-butanol is 3/1～5/1; By the polysaccharide solution after de-albumen with flowing water dialysis 18～24h, with deionized water dialysis 18～24h, again by 95% ethanol precipitation, the final concentration that makes polysaccharide is 50～100g/L, stand at low temperature 18～24h, the centrifugal 10～20min of 3000～5000rpm/min obtains sediment, collecting precipitation, through freeze drying 12～18h, obtain faint yellow polysaccharide powder;
The 3rd step, the CLA of take are core-clad material, Tween 80, class of department 80 are emulsifying agent, faint yellow polysaccharide powder and the gelatin of second step gained are made to microcapsule wall material by complex coacervation, and the mass ratio of core-clad material, wall material, emulsifying agent is 1:1:0.02, is prepared into the CLA microcapsules.
2. complex coacervation according to claim 1 prepares the CLA microcapsules that extracellular polysaccharide of bacteria is the wall material, it is characterized in that take that CLA is core-clad material, Tween 80, class of department 80 are emulsifying agent, extracellular polysaccharide of bacteria and gelatin are made microcapsule wall material by complex coacervation, the mass ratio of core-clad material, wall material, emulsifying agent is 1:1:0.02, and concrete operation method is:
By CLA, Tween 80, class of department 80 is added in gelatin solution while stirring, 800rpm/min, under 50 ℃, stir 30～40min and form stable pre-emulsion, then by polysaccharide solution at 500rpm/min, under 50 ℃, dropwise add in pre-emulsion, in the pre-emulsion formed, contain 98～102g/L CLA, 0.5～1.0g/L Tween 80, 0.5 class of～1g/L department 80, 48～52g/L gelatin and 48～52g/L polysaccharide, surplus is water, with 10% acetum, adjusting its pH is 3.2～3.8, reaction 1h, under 500rpm/min, make microcapsule emulsion be cooled to 1～10 ℃, freeze drying 12～18h, obtain exocellular polysaccharide CLA microcapsule product.
3. complex coacervation according to claim 1 prepares the CLA microcapsules that extracellular polysaccharide of bacteria is the wall material, it is characterized in that Tween 80, class 80 of department are emulsifying agent.
4. complex coacervation according to claim 1 prepares the CLA microcapsules that extracellular polysaccharide of bacteria is the wall material, it is characterized in that the microcapsules freeze drying optimum drying time is 12～18h.
5. complex coacervation according to claim 1 prepares the CLA microcapsules that extracellular polysaccharide of bacteria is the wall material, it is characterized in that above-mentioned bacterial isolates culture medium prescription is sucrose 25～40g/L, beef extract 10～30g/L, sodium chloride 5～30g/L, dipotassium hydrogen phosphate 0.5～1.5g/L, potassium dihydrogen phosphate 0.5～1.0g/L, magnesium sulfate 0.5～1.5g/L, manganese sulfate 0.5～1.0g/L, calcium chloride 0～0.5g/L, surplus is distilled water.
6. complex coacervation according to claim 1 prepares the CLA microcapsules that extracellular polysaccharide of bacteria is the wall material, it is characterized in that exocellular polysaccharide, as the microcapsules New Wall Material, is applied to food processing and preservation.
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| CN103897471A (en) * | 2014-02-24 | 2014-07-02 | 中南民族大学 | Linseed oil microcapsules capable of quickly forming films to timely repair scratches and preparation method thereof |
| CN104224751A (en) * | 2014-09-15 | 2014-12-24 | 青岛农业大学 | Preparation technology process of waxy corn nano starch embedded with conjugated linoleic acid |
| CN108559006A (en) * | 2018-04-20 | 2018-09-21 | 中国科学院青岛生物能源与过程研究所 | The method and its application of exocellular polysaccharide are prepared using the Microalgae fermentation waste liquid of production PUFA |
| CN114365742A (en) * | 2022-02-07 | 2022-04-19 | 吉林大学 | Protein/polysaccharide embedded origanum essential oil microcapsule and application thereof in fungus inhibition |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB1483303A (en) * | 1974-07-24 | 1977-08-17 | Fuji Photo Film Co Ltd | Encapsulation method |
| US20030193102A1 (en) * | 2002-04-11 | 2003-10-16 | Nianxi Yan | Encapsulated agglomeration of microcapsules and method for the preparation thereof |
| CN101445565A (en) * | 2008-12-24 | 2009-06-03 | 江南大学 | Separation and purification method for Dunaliella salina polysaccharide |
| CN101461784A (en) * | 2007-12-17 | 2009-06-24 | 黄云清 | Method for preparing nano magnetic microballoons and anticancer oral preparation prepared using the method |
| WO2009137964A1 (en) * | 2008-05-14 | 2009-11-19 | 陕西北美基因股份有限公司 | Preparation of superparamagnetic composite microparticles from cyclodextrin |
| CN101591859A (en) * | 2009-06-22 | 2009-12-02 | 华南农业大学 | A kind of Blumea oil microcapsule textile composite finishing agent and application thereof |
-
2013
- 2013-07-26 CN CN201310320692.3A patent/CN103406080B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB1483303A (en) * | 1974-07-24 | 1977-08-17 | Fuji Photo Film Co Ltd | Encapsulation method |
| US20030193102A1 (en) * | 2002-04-11 | 2003-10-16 | Nianxi Yan | Encapsulated agglomeration of microcapsules and method for the preparation thereof |
| CN101461784A (en) * | 2007-12-17 | 2009-06-24 | 黄云清 | Method for preparing nano magnetic microballoons and anticancer oral preparation prepared using the method |
| WO2009137964A1 (en) * | 2008-05-14 | 2009-11-19 | 陕西北美基因股份有限公司 | Preparation of superparamagnetic composite microparticles from cyclodextrin |
| CN101445565A (en) * | 2008-12-24 | 2009-06-03 | 江南大学 | Separation and purification method for Dunaliella salina polysaccharide |
| CN101591859A (en) * | 2009-06-22 | 2009-12-02 | 华南农业大学 | A kind of Blumea oil microcapsule textile composite finishing agent and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| 刘莎等: ""一株细菌ps-5胞外抗氧化性能研究多糖"", 《食品科技》, vol. 35, no. 6, 20 June 2010 (2010-06-20), pages 111 - 114 * |
| 王丽华等: "王丽华等", 《食品科技》, no. 1, 30 January 2003 (2003-01-30) * |
| 王兴敏等: ""以水溶性大豆多糖为壁材制备植酸酶微胶囊的研究"", 《食品科技》, vol. 33, no. 12, 20 December 2008 (2008-12-20), pages 56 - 59 * |
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| CN103897471A (en) * | 2014-02-24 | 2014-07-02 | 中南民族大学 | Linseed oil microcapsules capable of quickly forming films to timely repair scratches and preparation method thereof |
| CN104224751A (en) * | 2014-09-15 | 2014-12-24 | 青岛农业大学 | Preparation technology process of waxy corn nano starch embedded with conjugated linoleic acid |
| CN104224751B (en) * | 2014-09-15 | 2017-10-27 | 青岛农业大学 | Embed the preparation technology flow of the waxy corn nano-starch of CLA |
| CN108559006A (en) * | 2018-04-20 | 2018-09-21 | 中国科学院青岛生物能源与过程研究所 | The method and its application of exocellular polysaccharide are prepared using the Microalgae fermentation waste liquid of production PUFA |
| CN114365742A (en) * | 2022-02-07 | 2022-04-19 | 吉林大学 | Protein/polysaccharide embedded origanum essential oil microcapsule and application thereof in fungus inhibition |
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