Method for preparing beta-polymalic acid of high purity

A polymalic acid, high-purity technology, applied in the field of polymer preparation, can solve the problems of high cost, poor feasibility, complicated operation, etc., and achieve the effects of less equipment investment, simple method and low production cost

Inactive Publication Date: 2013-05-15
天津北洋百川生物技术有限公司
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

In the Chinese patent "Preparation Method of β-Polymalic Acid Simultaneously Obtaining By-product Pullulan (Publication No. CN101100687A)", the method introduced is to simultaneously obtain two products of polymalic acid and pullulan, and remove the melanin, but there is no specific mention of the purity of the polymalic acid obtained by this method
"Preparation method of β-polymalic acid and its salt (publication number is CN101487034A)" introduces that during fermentation, the fermentation broth is refluxed by membrane technology to separate high-molecular-weight polymalic acid, and low-molecular-weight malic acid is refluxed Continue to ferment in the fermentation broth, and use membrane separation method to further purify the sample. By this method, high molecular weight polymalic acid can be obtained, but the purity of the obtained polymalic acid is not mentioned, and medical polymalic acid Does not require too high a molecular weight
Moreover, this method is complicated to operate, poor in feasibility, and high in cost.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Prepare β-polymalic acid as follows:

[0029] 1. Preparation of culture medium

[0030] (1) Activation medium: potato dextrose medium (PDA): 200g / L potato, 20g / L glucose, 20g / L agar, pH6.0, sterilized by high pressure steam at 121℃ for 20min.

[0031] (2) Seed medium: glucose 80g / L, ammonium succinate 3g / L, succinic acid 2g / L, K 2 CO 3 0.4g / L, KH 2 PO 4 1g / L, MgSO 4 ·7H 2 O1g / L, ZnSO 4 ·7H 2 O5ppm, corn extract 10g / L, CaCO 3 20g / L, pH4.0-5.0, sterilized by high pressure steam at 121℃ for 20min.

[0032] (3) Fermentation medium: glucose 100g / L, sodium nitrate 20g / L, KH 2 PO 4 1g / L, MgSO 4 ·7H 2 O0.5g / L, peptone 20g / L, pH4.0-4.5, sterilized by high pressure steam at 121℃ for 20min, CaCO 3 20g / L, sterilized separately, sterilized by high pressure steam at 121℃ for 20min.

[0033] 2. Activation of strains

[0034] Aureobasidium pullulans TKPM00006 preserved in glycerol tubes was inoculated on the activation medium and cultured at 25°C for 2 days.

[0035] 3...

Embodiment 2

[0045] Prepare β-polymalic acid as follows

[0046] 1. Preparation of culture medium

[0047] (1) Activation medium: potato dextrose medium (PDA): 200g / L potato, 20g / L glucose, 20g / L agar, pH6.0, sterilized by high pressure steam at 121℃ for 20min.

[0048] (2) Seed medium: glucose 80g / L, ammonium succinate 3g / L, succinic acid 2g / L, K 2 CO 3 0.4g / L, KH 2 PO 4 1g / L, MgSO 4 ·7H 2 O1g / L, ZnSO 4 ·7H 2 O5ppm, corn extract 10g / L, CaCO 3 20g / L, pH4.0-5.0, sterilized by high pressure steam at 121℃ for 20min.

[0049] (3) Fermentation medium: glucose 100g / L, sodium nitrate 20g / L, KH 2 PO 4 1g / L, MgSO 4 ·7H 2 O0.5g / L, peptone 20g / L, pH4.0-4.5, sterilized by high pressure steam at 121℃ for 20min, CaCO 3 20g / L, sterilized separately, sterilized by high pressure steam at 121℃ for 20min.

[0050] 2. Activation of strains

[0051] Aureobasidium pullulans TKPM00006 preserved in glycerol tubes was inoculated on the activation medium and cultured at 23°C for 3 days.

[0052] 3....

Embodiment 3

[0062] Prepare β-polymalic acid as follows:

[0063] 1. Preparation of culture medium

[0064] (1) Activation medium: potato dextrose medium (PDA): 200g / L potato, 20g / L glucose, 20g / L agar, pH6.0, sterilized by high pressure steam at 121℃ for 20min.

[0065] (2) Seed medium: glucose 80g / L, ammonium succinate 3g / L, succinic acid 2g / L, K 2 CO 3 0.4g / L, KH 2 PO 4 1g / L, MgSO 4 ·7H 2 O1g / L, ZnSO 4 ·7H 2 O5ppm, corn extract 10g / L, CaCO 3 20g / L, pH4.0-5.0, sterilized by high pressure steam at 121℃ for 20min.

[0066] (3) Fermentation medium: glucose 100g / L, sodium nitrate 20g / L, KH 2 PO 4 1g / L, MgSO 4 ·7H 2 O0.5g / L, peptone 20g / L, pH4.0-4.5, sterilized by high pressure steam at 121℃ for 20min, CaCO 3 20g / L, sterilized separately, sterilized by high pressure steam at 121℃ for 20min.

[0067] 2. Activation of strains

[0068] Aureobasidium pullulans TKPM00006 preserved in glycerol tubes was inoculated on the activation medium and cultured at 25°C for 2 days.

[0069] 3...

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PUM

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Abstract

The invention discloses a method for preparing beta-polymalic acid of high purity, belonging to the field of the preparation of high-molecular polymer. The method for preparing beta-polymalic acid of high purity comprises the main steps of activation of strains, cultivation of seeds, fermentation culture and separation and purification, and is characterized in that the separation and purification method comprises the following steps of removing of pulan polysaccharide, extraction of rough polymalic acid, removing of polysaccharide difficult to separate in a sample, centrifugal collection of precipitate, and freeze drying. Compared with the existing invention, the method has the beneficial effects that firstly a sample of high purity can be obtained through a simpler method, fewer devices are invested, and the production cost is lower; and secondly the obtained sample has high purity and can lay a certain foundation for subsequent application.

Description

technical field [0001] The invention belongs to the field of polymer preparation, and the invention relates to a method for preparing β-polymalic acid, in particular to a method for preparing high-purity β-polymalic acid. Background technique [0002] β-polymalic acid (β-PMLA) is a polymer with malic acid as a monomer, because of its good water solubility, chemical derivatization, degradability, renewability, biological safety, etc. characteristics have attracted the attention of researchers. It is expected to be widely used in fields such as cosmetics, pharmaceuticals, and packaging materials. [0003] The production of polymalic acid mainly includes chemical synthesis method and microbial fermentation method. There are many chemical synthesis methods, which are mainly divided into direct polymerization method, lactide ring-opening method and lactone ring-opening method. Type and β type, because β type PMLA naturally exists in the organism, so the synthetic method of deve...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/62C08G63/90C12R1/645
Inventor 乔长晟姜少丽马正旺李政李振海钟海蛟
Owner 天津北洋百川生物技术有限公司
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