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Method for purifying pullulan polysaccharide

A pullulan polysaccharide and gel chromatography technology, applied in the biological field, can solve the problems of long reaction time and large solvent consumption, and achieve the effect of saving purification time

Active Publication Date: 2012-08-01
SHANDONG LUBEI PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The reaction time is long, dialysis is used many times, the solvent consumption is large, and the purity of the obtained pullulan is mostly between 90% and 95%.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] (a) crude product pullulan 100g adds distilled water and is diluted into 1.5% pullulan aqueous solution, adds chloroform / n-butanol (V 氯仿 :V 正丁醇 = 4: 1) mixed solution, fully shaken, centrifuged at 2000r / min for 15min, removed the precipitate, and then repeated the supernatant by this method 8 times, finally obtained the supernatant without protein, and removed the organic solution by distillation under reduced pressure , freeze-dried to obtain crude pullulan.

[0016] (b) Dissolve the crude pullulan in distilled water, add 95% ethanol to a concentration of 20% (volume concentration, the same below), ethanol analysis at 4°C for 8 hours, centrifuge at 15000r / min for 10min, and the precipitate is pullulan Polysaccharide, and then continue to add ethanol to the supernatant until the final concentrations are 30%, 40%, 50%, 60%, and 70%, respectively, and each operation is the same as described above.

[0017] (c) The crude pullulan obtained by fractional alcohol precipitat...

Embodiment 2

[0019] (a) 500g of crude product pullulan is diluted with distilled water into 2% pullulan aqueous solution, and chloroform / amyl alcohol (V 氯仿 :V 戊醇 =4:1) mixed solution, fully shaken, centrifuged at 3000r / min for 30min, removed the precipitate, and then repeated this method for supernatant 5 times, and finally obtained the supernatant without protein, and removed the organic solution by distillation under reduced pressure , freeze-dried to obtain crude pullulan.

[0020] (b) Dissolve the crude pullulan in distilled water, add 98% ethanol to a concentration of 15% (volume concentration, the same below), ethanol analysis at 4°C for 5 hours, centrifuge at 25000r / min for 10min, and the precipitate is pullulan Polysaccharides, and then continue to add ethanol to the supernatant to a final concentration of 30%, 45%, 60%, and 75%, respectively, and each operation is the same as described above.

[0021] (c) The crude pullulan obtained by fractional alcohol precipitation was separa...

Embodiment 3

[0023] (a) 200g of crude product pullulan is diluted with distilled water into 10% pullulan aqueous solution, adding chloroform / amyl alcohol (V 氯仿 :V 戊醇 =6:1) mixed solution, fully shaken, centrifuged at 4000r / min for 30min, removed the precipitate, and then repeated this method for 6 times on the supernatant, finally obtained the supernatant without protein, and removed the organic solution by distillation under reduced pressure , freeze-dried to obtain crude pullulan.

[0024] (b) Dissolve the crude pullulan polysaccharide in distilled water, add 98% ethanol to a concentration of 15% (volume concentration, the same below), ethanol analysis at 4°C for 10 hours, centrifuge at 20000r / min for 20min, and the precipitate is pullulan Polysaccharides, and then continue to add ethanol to the supernatant to a final concentration of 30%, 45%, 60%, and 75%, respectively, and each operation is the same as described above.

[0025] (c) The crude pullulan obtained by fractional alcohol p...

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PUM

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Abstract

The invention discloses a method for purifying pullulan polysaccharide. The method mainly comprises the following steps of: removing protein repeatedly for 4 to 20 times by using a Savage method, preparing a solution with different concentration gradient by using 95 percent or 98 percent ethanol, performing ethanol precipitation to remove small molecular substances, performing gel chromatography by using distilled water as an elution solvent, and thus obtaining purified pullulan polysaccharide of which the purity is over 99 percent and the yield is over 58 percent (based on polysaccharide content). The method has the advantages of short time, high yield, simplicity, high polysaccharide purity of the obtained pullulan polysaccharide and the like, and is suitable for industrialized production.

Description

technical field [0001] The invention relates to a method for purifying pullulan by using the Savage method, fractional alcohol precipitation and gel chromatography, and belongs to the field of biotechnology. technical background [0002] Pullulan is the Chinese translation of pullulan. It is an exopolysaccharide produced by the fermentation of Aureobasidium pullulans, a black yeast. It is a tasteless and odorless white powder. Its chemical composition is mainly polymaltotriose linked by α-1.4 glucosidic bonds. The molecular structure contains 1 / 3 of α-1.6 glucosidic bonds and 2 / 3 of α-1.4 glucosidic bonds. It is subject to different fermentation conditions effect, the molecular weight is generally at 5.0×10 4 -5.0×10 6 between Dalton. The polysaccharide is easily soluble in water, safe and non-toxic, edible, low calorific value, acid and alkali resistance, good plasticity, good film-forming property, good thin film gas barrier property. Because it can be degraded and uti...

Claims

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Application Information

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IPC IPC(8): C08B37/00
Inventor 王秀云肖佳普
Owner SHANDONG LUBEI PHARMA
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