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

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0014] Example 1

[0015] (a) Dilute 100g of crude pullulan with distilled water into a 1.5% aqueous solution of pullulan, add chloroform / n-butanol (V Chloroform : V N-butanol =4:1) The mixed solution, fully shaken, centrifuged at 2000r / min for 15min to remove the precipitate, the supernatant is processed again by this method for 8 times, and finally the protein-free supernatant is obtained, and the organic solution is removed by vacuum distillation , 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 hereinafter), alcohol precipitation at 4°C for 8 hours, centrifugation at 15000r / min for 10min, and the precipitate is pullulan Polysaccharides, and then continue to add ethanol to the supernatant until the final concentration is 30%, 40%, 50%, 60%, 70%, each operation is the same as described above.

[0017] (c) Separate the crude pullulan obtained by fractional ...

Example Embodiment

[0018] Example 2

[0019] (a) Dilute 500g of crude pullulan with distilled water into a 2% aqueous solution of pullulan, add chloroform / pentanol (V Chloroform : V Amyl alcohol =4:1) The mixed solution, fully shaken, centrifuged at 3000r / min for 30min, to remove the precipitate, the supernatant is processed again by this method for 5 times, and finally the protein-free supernatant is obtained, and the organic solution is removed by vacuum distillation , 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), alcohol precipitation at 4°C for 5 hours, centrifugation at 25000r / min for 10min, and the precipitate is pullulan Polysaccharide, and then continue to add ethanol to the supernatant until the final concentration is 30%, 45%, 60%, 75%, each operation is the same as the above.

[0021] (c) Separate the crude pullulan obtained by the fractional alcohol prec...

Example Embodiment

[0022] Example 3

[0023] (a) Dilute 200g of crude pullulan with distilled water into a 10% aqueous solution of pullulan, add chloroform / pentanol (V Chloroform : V Amyl alcohol =6:1) The mixed solution, fully shaken, centrifuged at 4000r / min for 30min to remove the precipitate, and the supernatant is treated with this method for 6 times, finally the protein-free supernatant is obtained, and the organic solution is removed by distillation under reduced pressure , Freeze-dried to obtain crude pullulan.

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

[0025] (c) Separate the crude pullulan obtained by fractional alcoh...

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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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