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Method for preparing beta-polymalic acid and simultaneously coproducing byproduct prussian blue

A technology of polymalic acid and by-products is applied in the field of biosynthesis of biodegradable polymer materials to achieve the effects of reducing environmental pollution, reducing production costs and avoiding environmental problems.

Inactive Publication Date: 2008-01-09
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, under the condition that the construction of high-yielding polymalic acid genetically engineered bacteria has not yet been realized, the removal of the by-product pullulan has always been one of the issues that require great attention in the production of polymalic acid

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Prepare β-polymalic acid and pullulan as follows:

[0024] The first step, the preparation of medium

[0025] ①Activation medium: potato sucrose medium (PSA): potato 200g / L, sucrose 20g / L, agar 20g / L, pH5.6, sterilized by high pressure steam at 121℃ for 30 minutes;

[0026] ② Preparation of fermentation medium: glucose 120g / L, NaNO 3 2g / L, KH 2 PO 4 0.1g / L, MgSO 4 ·7H 2 O 0.2g / L, KCl 0.5g / L, CaCO 3 30g / L, pH4.0, autoclaved at 121℃ for 15 minutes, CaCO 3 Dry heat sterilization alone, 180°C for 2 hours;

[0027] The second step, the activation of bacteria

[0028] Aureobasidium pullulans (Aureobasidium pullulans, As 3.837) stored in a glycerol tube at -70°C was activated and cultured at 24°C for 48 hours using the PSA medium prepared in the first step.

[0029] The third step, seed cultivation

[0030] Pick the single bacterium obtained in the second step on the solid plate, inoculate it into an L tube containing 5 mL of the fermentation medium prepared in the...

Embodiment 2

[0038] Prepare β-polymalic acid and pullulan as follows:

[0039] The first step, the preparation of medium

[0040]①Activation medium: potato sucrose medium (PSA): potato 200g / L, sucrose 20g / L, agar 20g / L, pH5.6, sterilized by high pressure steam at 121℃ for 30 minutes;

[0041] ② Preparation of fermentation medium: glucose 120g / L, NaNO 3 2g / L, KH 2 PO 4 0.1g / L, MgSO 4 ·7H 2 O 0.2g / L, KCl 0.5g / L, CaCO 3 30g / L pH4.0, autoclaved at 121℃ for 15 minutes, CaCO 3 Dry heat sterilization alone, 180°C for 2 hours;

[0042] The second step, the activation of bacteria

[0043] Aureobasidium pullulans (Aureobasidium pullulans, As 3.837) stored in a glycerol tube at -70°C was activated and cultured at 24°C for 48 hours using the PSA medium prepared in the first step.

[0044] The third step, seed cultivation

[0045] Pick the single bacterium obtained in the second step on the solid plate, inoculate it into an L tube containing 5 mL of the fermentation medium prepared in the ...

Embodiment 3

[0053] Prepare β-polymalic acid and pullulan as follows:

[0054] The first step, the preparation of medium

[0055] ①Activation medium: potato sucrose medium (PSA): potato 200g / L, sucrose 20g / L, agar 20g / L, pH5.6, 121℃, autoclaved for 30 minutes;

[0056] ② Preparation of fermentation medium: glucose 120g / L, NaNO 3 2g / L, KH 2 PO 4 0.1g / L, MgSO 4 ·7H 2 O 0.2g / L, KCl 0.5g / L, CaCO 3 30g / L, pH 4.0, autoclaved at 121°C for 15 minutes, CaCO 3 Dry heat sterilization alone, 180°C for 2 hours;

[0057] The second step, the activation of bacteria

[0058] Aureobasidium pullulans (Aureobasidium pullulans, As 3.837) stored in a glycerol tube at -70°C was activated and cultured at 22°C for 90 hours using the PSA medium prepared in the first step.

[0059] The third step, seed cultivation

[0060] Pick the single bacterium obtained in the second step on the solid plate, inoculate it into an L tube containing 5 mL of the fermentation medium prepared in the first step, and cultu...

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Abstract

Beta-polyoxysuccinic acid and Prussian blue as by-product are prepared by: inoculating activated fungus in fermenting medium for 8-14 days at 24-30 deg.C and 120rpm, separating solid from liquid to remove mycelia, adding alcohol with equal volume to separate Prussian blue deposit and melanochrome, concentrating supernatant and centrifuging with double-volume alcohol to separate Prussian blue, re-concentrating, adding four volume alcohol to deposit beta-polyoxysuccinic acid, and dissolving into water, removing residual melanochrome and micro-molecular materials, and frozen drying to obtain beta-polyoxysuccinic acid, dissolving separated Prussian blue in mixed alcohol-butanone to remove melanochrome, and dialyzing and frozen drying to obtain Prussian blue. It can remove melanochrome effectively to quality purity and reduce pollution by discharged Prussian blue and melanochrome.

Description

technical field [0001] The technical scheme of the invention belongs to the research field of biodegradable polymer materials synthesized by biological methods. Background technique [0002] The research on the synthesis of biodegradable polyester began in the 1960s. In 1969, Shimada and Matsushima, microbiologists who studied Penicillium cyclopium, speculated that there might be polymers containing malic acid structural units in the bacteria, which could inhibit the acid protease activity of Penicillium cyclopium, and finally The polymer was confirmed to be polymalic acid (PMA). In 1979, Vert et al first synthesized a water-soluble aliphatic polyester - polymalic acid, which uses malic acid as the only monomer. In 1989, Fisher et al. isolated PMA from Physarum Polycephalum cells and found that it was an inhibitor of DNA polymerase α. So far, foreign scholars represented by Vert, Cammas, Kajiyama, etc. have done in-depth research on the synthesis, performance and applicat...

Claims

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Application Information

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IPC IPC(8): C12P19/04C12P1/02C12R1/645
Inventor 宋存江孙村民马晨燕郭红彦刘莉刘娜王淑芳
Owner NANKAI UNIV
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