Preparation f beta-poly malic acid and salt thereof

A technology of polymalic acid and malate, which is applied in the field of fermentation engineering, can solve the problems of no large-scale application of β-polymalic acid, high production cost, and long fermentation cycle, so as to shorten the fermentation cycle, increase yield, Reduces the effect of feedback inhibition

Active Publication Date: 2009-07-22
INST OF PROCESS ENG CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] So far, there is no large-scale application of fermentation to produce β-polymalic acid. The main reasons are: the fermentation period is too long, the yield is low, the production cost is too high, and the β-polymalic acid produced is a variety of polymers with different degrees of polymerization. Complex

Method used

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  • Preparation f beta-poly malic acid and salt thereof
  • Preparation f beta-poly malic acid and salt thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Prepare β-polymalic acid, β-polymalate and pullulan as follows:

[0049] The first step, culture medium preparation

[0050] ①Activation medium: potato dextrose medium, sterilized at 121°C for 15 minutes;

[0051] ② Preparation of fermentation medium (w / v): 10% glucose, 0.2% NaNO 3 , 0.01% KH 2 PO 4 , 0.02% MgSO 4 ·7H 2 O, 0.05% KCl, pH 4.0-4.5, sterilized at 121°C for 15 minutes.

[0052] The second step, the activation of bacteria

[0053] The strain of Aureobasidium pullulans ipe-1 stored at 4°C was transferred to the slant of the activation medium, and the activation culture was carried out at 25°C for 48 hours.

[0054] The third step, seed cultivation

[0055] Inoculate the activated seed culture on the slant into a 500ml Erlenmeyer flask containing 100ml of fermentation medium, and culture it with shaking at 25°C and 200r / min for 48 hours.

[0056] The fourth step, inoculation and fermentation

[0057] The culture obtained in the third step was inoculat...

Embodiment 2

[0063] Prepare β-polymalic acid, β-polymalate and pullulan as follows:

[0064] The first step, culture medium preparation

[0065] Same as Example 1

[0066] The second step, the activation of bacteria

[0067] Same as Example 1

[0068] The third step, seed cultivation

[0069] Same as Example 1

[0070] The fourth step, inoculation and fermentation

[0071] Inoculate the culture obtained in the third step into a 7L fermenter with 4L of fermentation medium at an inoculum size of 10%, with an initial pH of 4.0-4.5, a temperature of 25°C, an aeration rate of 0.5-2.5vvm, and a rotation speed of 600-700r / min, cultivated for 7 days, at this time, the bacteria were partially autolyzed, and the sugar consumption rate decreased. Finally, the content of β-polymalic acid in the fermented liquid was 8.7g / L, and the molecular weight was in a wide range of 5KDa-1000kDa.

[0072] Step 5 Preparation of β-polymalic acid and by-product pullulan

[0073] ① Centrifuge the β-polymalic ...

Embodiment 3

[0077] Prepare β-polymalic acid, β-polymalate and pullulan as follows:

[0078] The first step, culture medium preparation

[0079] Same as Example 1

[0080] The second step, the activation of bacteria

[0081] Same as Example 1

[0082] The third step, seed cultivation

[0083] Same as Example 1

[0084] The fourth step, inoculation and fermentation

[0085]Inoculate the culture obtained in the third step into the fermentation medium, pH 4.0-4.5, temperature 25°C, air flow 0.5-2.5vvm, rotation speed 600-700r / min, cultivate for 72h, start pump 1 to make the cells pass through Membrane module A (see attached picture, membrane pore size 0.8μm) is backflowed into the fermenter, the pressure difference is 0.05-0.1mpa, the permeate 1 is concentrated by membrane module B (see attached picture, molecular weight cut-off 100KDa), and the concentrated solution is returned to the storage tank 1. The permeate 2 is returned to the fermenter. Control the content of β-polymalic acid ...

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Abstract

The invention discloses a method for preparing Beta-poly malic acid and a salt thereof and simultaneously obtaining a byproduct pullulan. The method is realized by adopting the following technical proposal: return thalli and interception are implemented by a membrane technique in a fermentation process, high molecular weight Beta-poly malic acid is concentrated, and low molecular weight Beta-polymalic acid, pullulan and other components are recycled in a fermentation system; a weakly basic anion exchange resin is adopted to absorb the Beta-poly malic acid and lead pullulan solution to penetrate, alkaline solution is adopted to elute the Beta-poly malic acid that is adsorbed on the resin, and eluting solution is purified and dried, thus obtaining the salt product of the Beta-poly malic acid; a penetrating fluid is concentrated, chloroform and butanone mixture solution is adopted to remove protein, and the pullulan is precipitated by ethanol alcohol precipitation, orderly washed by acetone and ethyl ether, and dried by P2O5, thus obtaining a powder pullulan product; a strong acid cation exchange resin is adopted to absorb salt ions in salt solution of the Beta-poly malic acid, a membrane is used for removing extra acid in the desalted solution, and the obtained solution is concentrated and dried, thus obtaining the Beta-poly malic acid product.

Description

technical field [0001] The technical scheme of the present invention belongs to the technical field of fermentation engineering, and specifically relates to the use of membrane technology to reflow bacteria, intercept and concentrate β-polymalic acid and control the degree of polymerization of β-polymalic acid, and use resin to prepare β-polymalate, β-polymalic acid Methods of polymalate and pullulan. Background technique [0002] 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. In 1979, Vert et al first synthesized a water-soluble aliphatic polyester - polymalic acid, which uses polymalic acid as the only monomer. In 1989, Fisher et al. isolated polymalic acid from Physarum Polycephalum cells and found that it wa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/04C12P1/02C08B37/00C08G63/06C12R1/645
Inventor 万印华曹伟锋苏仪沈飞陈向荣赵方
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
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