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Method for producing polymalic acid through repeated fermentation that can be made into resting cells

A technology of resting cells and polymalic acid is applied in the field of fermentation and production of polymalic acid, which can solve the problems of difficulty in obtaining polymalic acid and high production cost, and achieve the effects of increasing fermentation yield, improving fermentation efficiency and reducing production cost.

Inactive Publication Date: 2011-12-21
天津实发中科百奥工业生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Because the production cost is very high, synthetic polymalic acid and natural polymalic acid are still relatively difficult to obtain, so it is important to provide an efficient, low-cost method for producing polymalic acid through repeated fermentation of resting cells. One of the new topics urgently needed to be developed by researchers in the technical field

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] (1) Preparation of slant strains

[0020] Prepare the slant medium according to the following ratio:

[0021] Potato: 200g / L,

[0022] Glucose: 20g / L,

[0023] Agar: 20g / L,

[0024] Use 10% sodium hydroxide to adjust the pH to 5.6; after sterilizing at 115°C for 15 minutes to make a slant, pick a single colony from the plate and inoculate it by streaking, that is, inoculate Aureobasidium pullulans TJZKBIO153 on the sterile slant, and keep the temperature at 30°C After 48 hours of cultivation, black spores appeared.

[0025] (2) Preparation of seed solution

[0026] Prepare the seed medium according to the following ratio:

[0027] Glucose: 60g / L,

[0028] Ammonium succinate: 3g / L,

[0029] Corn syrup: 0.5g / L,

[0030] Succinic acid: 2g / L,

[0031] Potassium dihydrogen phosphate: 0.1g / L,

[0032] Sodium carbonate: 0.4g / L,

[0033] Magnesium sulfate: 0.1g / L,

[0034] Zinc sulfate: 0.005g / L,

[0035] Use 10% sodium hydroxide to adjust the pH to 5.0, sterilize ...

Embodiment 2

[0069] (1) Preparation of slant strains

[0070] Prepare the slant medium according to the following ratio:

[0071] Potato: 200g / L,

[0072] Glucose: 20g / L,

[0073] Agar: 20g / L,

[0074] Use 10% sodium hydroxide to adjust the pH to 5.6; after sterilizing at 115°C for 15 minutes to make a slant, pick a single colony from the plate and inoculate it by streaking, culture at a constant temperature of 28°C for 60 hours, and black spores appear.

[0075] (2) Preparation of seed solution

[0076] Prepare the seed medium according to the following ratio:

[0077] Glucose: 80g / L,

[0078] Ammonium succinate: 3.5g / L,

[0079] Peptone: 1.5g / L,

[0080] Maleic acid: 5g / L,

[0081] Potassium dihydrogen phosphate: 0.1g / L,

[0082] Sodium carbonate: 0.4g / L,

[0083] Magnesium sulfate: 0.1g / L,

[0084] Zinc sulfate: 0.005g / L,

[0085] Use 10% sodium hydroxide to adjust the pH to 5.5, sterilize at 115°C for 15 minutes, and when the temperature drops to 28°C, insert the slant bact...

Embodiment 3

[0112] (1) Preparation of slant strains

[0113] Prepare the slant medium according to the following ratio:

[0114] Potato: 200g / L,

[0115] Glucose: 20g / L,

[0116] Agar: 20g / L,

[0117] Use 10% sodium hydroxide to adjust the pH to 5.6; after sterilizing at 115°C for 15 minutes to make a slant, pick a single colony from the plate and inoculate it by streaking, culture at a constant temperature of 30°C for 50 hours, and black spores appear.

[0118] (2) Preparation of seed solution

[0119] Prepare the seed medium according to the following ratio:

[0120] Glucose: 80g / L,

[0121] Ammonium succinate: 3.0g / L,

[0122] Peptone: 0.5g / L,

[0123] Succinic acid: 2g / L,

[0124] Potassium dihydrogen phosphate: 0.1g / L,

[0125] Sodium carbonate: 0.4g / L,

[0126] Magnesium sulfate: 0.1g / L,

[0127] Zinc sulfate: 0.005g / L,

[0128] Use 10% sodium hydroxide to adjust the pH to 5.3, sterilize at 115°C for 15 minutes, and when the temperature drops to 30°C, insert the slant bact...

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Abstract

The invention relates to a method for producing polymalic acid through continuous fermentation which can be made into resting cells. The implementation steps are as follows: (1) preparation of slant strains; (2) preparation of seed liquid; (3) the above seed liquid is used for inoculation into Fermentation in a 5L aseptic fermenter; (4) Do not control the pH in the early stage of fermentation, and then continuously add NaOH to control the pH of the fermentation broth to 5.0-5.5 after the emergence of mycelium balls; (5) After 55-60 hours of fermentation, the When the glucose concentration is 8-10g / L, stop the fermentation, add fresh medium again, and repeat the fermentation; (6) Resting cell preparation; (7) Use the same fermentation method for resting cell fermentation; (8) Fermentation The solution was filtered with a hollow fiber membrane with a molecular weight cut-off of 6000Da, added an equal volume of absolute ethanol to precipitate, dried and weighed; the yield of polymalic acid was 50-57g / L. The method can effectively reduce the non-optimal economic stage in the fermentation process, and achieve the purposes of shortening the fermentation time, improving the utilization rate of equipment, and reducing the production cost of products.

Description

technical field [0001] The invention relates to a method for producing polymalic acid by fermentation, in particular to a method for repeatedly fermenting and producing polymalic acid which can be made into resting cells. Background technique [0002] Polymalic acid (PMLA) is a polymer with excellent biocompatibility, biodegradability and bioabsorbability. It was originally discovered by Shimada et al. in 1969 when they studied a Penicillium arcuum. As a water-soluble aliphatic polyester, it has high water solubility and chemical derivatization. Polymalic acid can be degraded spontaneously or enzymatically in aqueous solution to generate small molecule L-malic acid. The L-malic acid monomer can be absorbed by the human body. Absorption does not have any side effects. After polymalic acid is decomposed and burned, the final products are carbon dioxide and water, which can be absorbed by plants and are non-toxic and harmless to the environment. The many properties of polymal...

Claims

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Application Information

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IPC IPC(8): C12P7/62C12R1/645
Inventor 刘云徐勇虎李睿颖国华胡娅君王永乐王晓琼范婷
Owner 天津实发中科百奥工业生物技术有限公司
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