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Method for preparing L-hydroxyproline through enzymolysis

A technology of hydroxyproline and enzymatic hydrolysis, which is applied in the field of enzymatic hydrolysis to prepare L-hydroxyproline, can solve the problems of easy oxidation of hydroxyproline, large amount of acid and alkali, and unstable yield, so as to improve the fermentation level , reduce the loss of raw materials, reduce the effect of production costs

Inactive Publication Date: 2019-07-05
南通普悦生物医药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method hydroxyproline is easy to oxidize, and the yield is unstable; the water consumption is large, the amount of acid and alkali is large, and a large amount of waste residue and waste water will be produced in the production process, which will cause very serious pollution to the environment.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] The method for enzymatic hydrolysis method L-hydroxyproline, comprises the following steps:

[0022] S1, the culture tank is sterilized first, then the liquid medium is put into the culture tank, and then inoculated;

[0023] S2. Insert the L-proline producing strain into the culture tank, the inoculation temperature is 30°C, and the pH is 7.0. Insert the needle of the inoculation bottle into the inoculation port. After the inoculation, place a cotton ball soaked in 75% ethanol in On the inoculation port, pull out the inoculation needle and start culturing;

[0024] S3. Cultivation: culture temperature is 30°C, stirring speed is 200rpm, culture time is 8 hours, when the dissolved oxygen in the fermentation process rises to 10%, start to add glucose, control the dissolved oxygen in the fermentation process to 10%, and the final glucose content is less than 10g / L, the strains are cultivated in liquid medium to obtain mature seeds;

[0025] S4, the mature seeds are move...

Embodiment 2

[0032] The method for enzymatic hydrolysis method L-hydroxyproline, comprises the following steps:

[0033] S1, the culture tank is sterilized first, then the liquid medium is put into the culture tank, and then inoculated;

[0034] S2. Insert the L-proline production strain into the culture tank, the inoculation temperature is 32°C, and the pH is 8.0. Insert the needle of the inoculation bottle into the inoculation port. After the inoculation, place a cotton ball soaked in 75% ethanol by volume On the inoculation port, pull out the inoculation needle and start culturing;

[0035] S3. Cultivation: culture temperature is 32°C, stirring speed is 250rpm, culture time is 15 hours, when the dissolved oxygen in the fermentation process rises to 20%, start to add glucose, control the dissolved oxygen in the fermentation process to 20%, and the final glucose content is less than 10g / L, the strains are cultivated in liquid medium to obtain mature seeds;

[0036] S4, the mature seeds...

Embodiment 3

[0044] The method for enzymatic hydrolysis method L-hydroxyproline, comprises the following steps:

[0045] S1, the culture tank is sterilized first, then the liquid medium is put into the culture tank, and then inoculated;

[0046] S2. Insert the L-proline production strain into the culture tank, the inoculation temperature is 35°C, and the pH is 9.0. Insert the needle of the inoculation bottle into the inoculation port. After the inoculation, place a cotton ball soaked in 75% ethanol in On the inoculation port, pull out the inoculation needle and start culturing;

[0047]S3. Cultivation: culture temperature is 35°C, stirring speed is 300rpm, culture time is 24 hours, when the dissolved oxygen in the fermentation process rises to 30%, start to add glucose, control the dissolved oxygen in the fermentation process to 30%, and the final glucose content is less than 10g / L, the strains are cultivated in liquid medium to obtain mature seeds;

[0048] S4, the mature seeds are mov...

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Abstract

The invention discloses a method for preparing L-hydroxyproline through enzymolysis. The method comprises inoculating L-proline producing strains into a culture tank, during fermentation, when the content of dissolved oxygen increases to 10-30%, batch-feeding glucose to control the content of dissolved oxygen during fermentation to be 10-30% and the final content of glucose to be lower than 10 g / L, and culturing the strains in fluid mediums to obtain mature seeds; transferring the mature seeds into the fermentation mediums of a fermentation tank for fermentation metabolism to produce L-hydroxyproline, micro-filtering L-hydroxyproline fermentation liquor to obtain micro-filtered clear liquor and microfiltered retained liquor, performing adsorption with highly acidic cation exchange resin and then weakly alkaline anion exchange resin to obtain ion exchange liquor; obtaining the finished product of the L-hydroxyproline. By means of a DO-stat (dissolved oxygen-static) fed-batch culture control method, repression effects of the glucose during fermentation production of L-proline can be effectively eliminated, and the average yield of the L-proline can be increased; the microfiltered retained liquor and superfluous mother liquor are returned to the ion exchange resin adsorption processes, so that loss of raw materials can be effectively reduced, the fermentation level can be increased, the production cost can be reduced and three-waste pollution can be avoided.

Description

technical field [0001] The invention relates to a method for L-hydroxyproline, in particular to a method for preparing L-hydroxyproline by enzymatic hydrolysis. Background technique [0002] Hydroxyproline (Hyp) is an imino acid, which is the product of L-proline hydroxylation, and its molecular formula is C 5 h 9 NO 3 . Hydroxyproline is white flaky crystal or crystalline powder, slightly sweet, with a melting point of 274°C, easily soluble in water and slightly soluble in ethanol. There are three main production methods of L-hydroxyproline: chemical method, hydrolysis extraction method and microbial fermentation method. Among them, the process of producing L-proline by chemical method and hydrolysis extraction method is complicated, with low benefit and high cost. Microbial fermentation method is increasingly dominant in industrial production due to its advantages of low cost, less pollution, less impurities and high purity. [0003] The separation and extraction of h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/24C07D207/16C12R1/15
CPCC12P13/24C07D207/16
Inventor 王林
Owner 南通普悦生物医药有限公司
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