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Method for improving the fermentation yield of hyaluronic acid (HA)

A hyaluronic acid and production technology, applied in the biological field, can solve the problems of physiological and metabolic characteristics, production can not be further improved, constraints, etc., to achieve the effect of increasing fermentation production

Inactive Publication Date: 2013-09-25
SICHUAN HENGYI TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a method for improving the production of HA by microbial fermentation and reducing the production cost, which is restricted by the physiological and metabolic characteristics of the production strain Streptococcus zooepidemicus. A kind of method that improves hyaluronic acid fermentation yield
The present invention subverts the traditional fermentation culture method, and effectively solves the problem that the output cannot be further improved when the production strain Streptococcus zooepidemicus is used to produce HA. The present invention is important for improving the fermentation output of hyaluronic acid, reducing the production cost of enterprises, and promoting the application of HA. significance

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] 1. Preparation

[0032] 1) Prepare buffer

[0033] Each component was weighed according to the following parts by weight ratio: 36.76 parts of disodium hydrogen phosphate, 15.98 parts of sodium dihydrogen phosphate, and 12.5 parts of sodium bicarbonate, and the components were mixed to prepare a buffer solution. Each liter of buffer contains 36.76g disodium hydrogen phosphate, 15.98g sodium dihydrogen phosphate, and 12.5g sodium bicarbonate.

[0034] 2) Preparation of trace element solution

[0035] Each component was weighed according to the following ratio of parts by weight: 2.0 parts of calcium chloride, 0.046 part of zinc chloride, and 0.019 part of copper sulfate pentahydrate, and each component was mixed with water to prepare a trace element solution. Each liter of trace element solution contains 2.0g of calcium chloride, 0.046g of zinc chloride, and 0.019g of copper sulfate pentahydrate.

[0036] 3) Preparation of slant medium

[0037] Each component was wei...

Embodiment 2

[0053] 1. Preparation

[0054] The preparation of buffer solution, preparation of trace element solution, slant culture medium, seed culture medium and fermentation medium is all the same as in Example 1.

[0055] 2. Preparation

[0056] (1) Slant culture: inoculate Streptococcus zooepidemicus strains on the slant culture medium, place the slant medium in a constant temperature incubator and cultivate for 12 hours, and the temperature in the constant temperature incubator is 40°C to obtain slant seeds.

[0057] (2) Seed cultivation: Inoculate the prepared slant seeds into a 2L Erlenmeyer flask containing 500mL of seed medium for cultivation. When the pH value of the base is reduced to 5.6-6.0, the seed liquid is obtained.

[0058] (3) Fermentation culture: Put the seed liquid into a fermenter with fermentation medium, the stirring speed is 120 r / min, the ventilation rate is 100 vvm, the tank pressure is 0.03 Mpa, and the temperature is 37°C, and then the first stage of culti...

Embodiment 3

[0066] 1. Preparation

[0067] The preparation of buffer solution, preparation of trace element solution, slant culture medium, seed culture medium and fermentation medium is all the same as in Example 1.

[0068] 2. Preparation

[0069] (1) Slant culture: inoculate Streptococcus zooepidemicus strains on the slant culture medium, place the slant medium in a constant temperature incubator and cultivate for 16 hours, and the temperature in the constant temperature incubator is 35°C to obtain slant seeds.

[0070] (2) Seed culture: Inoculate the prepared slant seeds into a 2L Erlenmeyer flask containing 500mL of seed medium for culture. When the pH value of the base is reduced to 5.6-6.0, the seed liquid is obtained.

[0071] (3) Fermentation culture: Put the seed liquid into a fermenter with fermentation medium, the stirring speed is 120 r / min, the ventilation rate is 100 vvm, the tank pressure is 0.03 Mpa, and the temperature is 37°C, and then the first stage of cultivation i...

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Abstract

The invention discloses a method for improving the fermentation yield of hyaluronic acid (HA), and aims to solve the problems that the yield of the HA is restricted under the influence of the physiological metabolic properties of streptococcus zooepidemicus serving as production strains, while the yield of the HA produced by a microbial fermentation method is required to be improved and the production cost is required to be reduced imperatively at present. The method comprises the following steps: slant culture, seed culture, fermentation culture, first-stage culture and second-stage culture. By utilizing the method, the fermentation yield of the HA can be further improved and the fermentation comprehensive cost is reduced in the mode of combining fed-batch culture and intermittent pH stress, so that the application of the HA in the field of medicaments, makeup and food healthcare is promoted. By utilizing the method, the conventional fermentation culture mode is subverted; the problem that the yield cannot be further improved in the method for producing the HA is effectively solved; the method has important significance to improve the fermentation yield of the HA, reduce the production cost of enterprises and promote the application of the HA.

Description

technical field [0001] The invention relates to the field of biology, in particular to a method for increasing the fermentation yield of hyaluronic acid. During the production of hyaluronic acid by fermentation of Streptococcus zooepidemicus, the combination of fed-batch culture and interstitial pH stress can effectively improve the yield of hyaluronic acid. Fermentation yield of hyaluronic acid. Background technique [0002] Hyaluronic acid, HA for short, is an acidic mucopolysaccharide formed by linking glucuronic acid and acetylglucosamine through β-(1→3) and β-(1→4) glycosidic bonds. It has excellent properties such as high moisture retention, viscoelasticity and biocompatibility, especially its unique moisture retention performance. It has been widely used in medicine, daily chemical, food and other fields, and is one of the hot products in the biochemical industry at home and abroad. . [0003] In recent years, microbial fermentation has replaced animal tissue extrac...

Claims

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Application Information

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IPC IPC(8): C12P19/26C12R1/46
Inventor 邱平柯常毅李涛
Owner SICHUAN HENGYI TECH
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