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Process for producing beta-polymalic acid by feed-batch fermentation

A technology of polymalic acid and fed-feed fermentation, which is applied in the directions of fermentation, microorganism-based methods, microorganisms, etc., can solve the problems such as the unreported process of fed-feed fermentation, and achieve the effects of reducing costs and improving fermentation yield.

Inactive Publication Date: 2011-08-17
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

[0007] In the current polymalic acid fermentation process, there is no report on the process of fed-batch fermentation

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Produce β-polymalic acid by fermentation as follows:

[0030] The first step, culture medium preparation

[0031] ①Activation medium: potato dextrose medium, sterilized at 121°C for 15 minutes;

[0032] ② Preparation of seed medium (w / v): 8% glucose, 0.2% sodium nitrate, 0.01% potassium dihydrogen phosphate, 0.02% magnesium sulfate heptahydrate, 0.05% potassium chloride, 0.1% peptone, pH 4.0-4.5, Sterilize at 121°C for 15 minutes.

[0033] ③ Preparation of fermentation medium (w / v): 100-300g / L glucose, 6g / L sodium nitrate, 0.1-0.6g / L potassium dihydrogen phosphate, 0.2g / L magnesium sulfate heptahydrate, 0.5g / L chlorine Potassium chloride, 15g / L peptone, 5g / L yeast powder, 0.005-0.1g / L zinc sulfate heptahydrate, 0.04-1.0g / L calcium chloride, pH4.0-7.0, sterilized at 121°C for 15min.

[0034] The second step, the activation of bacteria

[0035] The strain of Aureobasidium pullulans ipe-1 stored at 4°C was transferred to the slant of the activation medium, and the acti...

Embodiment 2

[0041] Produce β-polymalic acid by fermentation as follows:

[0042] The first step, culture medium preparation

[0043] Same as Example 1

[0044] The second step, the activation of bacteria

[0045] Same as Example 1

[0046] The third step, seed cultivation

[0047] Same as Example 1

[0048] The fourth step, inoculation and fermentation

[0049]Inoculate the culture obtained in the third step into a 7L fermenter with 4L fermentation medium at an inoculum size of 10%. -0.1Mpa, control the dissolved oxygen concentration to be greater than 70% 16 hours before fermentation, then control the dissolved oxygen concentration at 70%, and cultivate for 72 hours. The initial sugar is 190g / L glucose or 190g / L sucrose, the fermentation period is 60h and 72h respectively, the content of β-polymalic acid in the fermentation broth is 43.8g / L and 42.3g / L respectively, and the dry weight of the bacteria is 68.9g respectively / L and 70.2g / L.

Embodiment 3

[0051] Produce β-polymalic acid by fermentation as follows:

[0052] The first step, culture medium preparation

[0053] Same as Example 1

[0054] The second step, the activation of bacteria

[0055] Same as Example 1

[0056] The third step, seed cultivation

[0057] Same as Example 1

[0058] The fourth step, inoculation and fermentation

[0059] Inoculate the culture obtained in the third step into a 7L fermenter with 4L fermentation medium at an inoculum size of 10%. -0.1Mpa, control the dissolved oxygen concentration greater than 70% 16 hours before fermentation, and then control the dissolved oxygen concentration at 70%. The initial sugar is 190g / L glucose, the feed solution fed at the beginning of fermentation 36h contains 300g / L or 800g / L glucose, the fermentation cycle is 72h, the feed solution 1L is added continuously, and the β-polymalic acid in the fermentation liquid is The contents were 38.6g / L and 45.3g / L respectively, and the dry weights of bacteria wer...

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Abstract

The invention discloses a feed-batch fermentation process improving the yield of beta-polymalic acid, which is implemented by the following scheme: culturing a strain aureobasidium pullulans ipe-1 in a culture medium containing a carbon source, a nitrogen source, inorganic salt and trace elements while controlling the pH value and dissolved oxygen; and based on a batch fermentation production process, adopting glucose as the initial sugar, and 36-48 hours later, starting to continuously add the feed-batch solution of which the carbon source is glucose, sucrose, a mixed solution of glucose and fructose or a mixed solution of glucose and sucrose, and the inorganic salt contains sodium nitrate, calcium chloride and zinc sulfate. The yield of beta-polymalic acid is improved by 20-50% in comparison with the batch fermentation process. Through the invention, the original production method is changed, the yield of the beta-polymalic acid is obviously improved, the cost is lowered, and the process disclosed by the invention can be used for industrial large-scale fermentation production of beta-polymalic acid.

Description

technical field [0001] The technical scheme of the invention belongs to the technical field of fermentation engineering, and specifically relates to a method for improving the fermentation production of β-polymalic acid by a strain of Aureobasidium pulluluns (A. pulluluns ipe-1) by using a feeding process. Background technique [0002] Polymalic acid is a special kind of aliphatic polyester, which is composed of malic acid as the only monomer and linked by ester bonds. Different from many other natural multipolymers, there are many free carboxyl groups and asymmetric carbon atoms in polymalic acid molecules. These free carboxyl groups endow it with many special properties. In addition to its good water solubility, biodegradability, biophase In addition to capacity and bioabsorbability, it also has two significant advantages: (1) easy metabolism. Since L-malic acid is an intermediate of the Krebs tricarboxylic acid cycle in the organism, β-polymalic acid is easily removed in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/62C12R1/645
Inventor 万印华曹伟锋苏仪陈向荣沈飞
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
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