CN1786180A - Method of improving byaluronic acid yield and molecular weight by fermentation process - Google Patents

Abstract

The invention discloses a method for increasing the yield and molecular weight of hyaluronic acid produced by fermentation, and relates to the technical field of fermentation for producing hyaluronic acid. The present invention uses Streptococcus zooepidemicus H23 as the starting strain to produce hyaluronic acid through liquid fermentation. The fermentation medium uses yeast powder as the single nitrogen source and the stirring speed is 180-220rpm. And the molecular weight is greatly improved, which is suitable for the industrialized production of hyaluronic acid. The invention improves the output and molecular weight of the hyaluronic acid produced by the fermentation method, saves energy consumption in industrial production, reduces the production cost, and improves the quality of the hyaluronic acid product.

Description

A method for improving the yield and molecular weight of hyaluronic acid produced by fermentation

technical field

The invention relates to a method for increasing the yield and molecular weight of hyaluronic acid produced by fermentation, and relates to the technical field of fermentation for producing hyaluronic acid.

Background technique

Hyaluronic acid (HA), also known as hyaluronic acid or hyaluronic acid, is a macromolecular mucopolysaccharide widely present in the connective tissue of organisms. The physiological functions of HA are different in different tissues, such as water retention in the skin, lubrication in the synovial fluid of the joints, and permeability regulation in the blood vessel wall. In addition, as a polyanion electrolyte, HA has a large number of negative charges on the molecule, which can adjust the concentration of positive and negative ions around it and inhibit the activity of various enzymes. It is widely used in ophthalmic viscous surgery, treatment of joint diseases, soft tissue repair and as a drug carrier , especially the prevention and reduction of tissue adhesions after surgery has made great progress.

The production of hyaluronic acid by microbial fermentation has greatly expanded the source of HA and promoted the research and application of HA. The traditional fermentation method for producing hyaluronic acid mostly uses yeast powder and beef extract as nitrogen source, and adopts high stirring speed. The yield and molecular weight of hyaluronic acid obtained are low, which affects the quality of the final hyaluronic acid product.

Contents of the invention

The purpose of the present invention is to provide a method for improving the yield and molecular weight of hyaluronic acid produced by fermentation and simplify the production process to meet the needs of industrial production. The invention can improve the fermentation process of traditional fermentation method to produce hyaluronic acid, increase the yield and molecular weight of hyaluronic acid, and improve the quality of hyaluronic acid products.

Technical scheme of the present invention: adopt Streptococcus zooepidemicus (Streptococcus zooepidemicus) H23 as starting bacterial strain, this bacterial classification has been disclosed in March, 2003 in the 3rd phase of "Journal of Chemical Industry" volume 54, P350-356, by Jiangnan University Bioengineering Provided by the college; hyaluronic acid is produced by liquid fermentation. The fermentation medium uses 8.0-12.0g/L yeast powder as the single nitrogen source, and the stirring speed is 180-220rpm during the fermentation process to increase the production and molecular weight of hyaluronic acid.

The composition of the fermentation medium used in the production of hyaluronic acid by fermentation method is: glucose 65.8g/L, yeast powder 8.0-12.0g/L, Na ₂ HPO ₄ 12H ₂ O 6.2g/L, K ₂ SO ₄ 1.3g/L, MgSO ₄ ·7H ₂ O 2.0g/L, FeSO ₄ ·7H ₂ O 0.005g/L; the process conditions are: temperature 37°C, ventilation rate 1.2vvm, optimum stirring speed 200rpm.

Beneficial effects of the present invention: the fermentation process uses a lower stirring speed, which greatly increases the yield and molecular weight of hyaluronic acid, which is beneficial to energy saving and consumption reduction in industrial production. The method of the present invention improves the traditional fermentation method for producing hyaluronic acid. Using yeast powder alone as a nitrogen source to reduce production costs is of great significance to industrial production.

Detailed ways

Example 1

Streptococcus zooepidemicus (Streptococcus zooepidemicus) H23 is used as the starting strain to produce hyaluronic acid through liquid fermentation. The fermentation medium uses 8.0-12.0g/L yeast powder as the single nitrogen source, and the composition of the fermentation medium is: glucose 65.8g/L, Yeast powder 8.0～12.0g/L, Na ₂ HPO ₄ 12H ₂ O 6.2g/L, K ₂ SO ₄ 1.3g/L, MgSO ₄ 7H ₂ O 2.0g/L, FeSO ₄ 7H _{2 O} 0.005g /L; the process conditions are: temperature 37°C, ventilation rate 1.2vvm, stirring speed around 200rpm during the fermentation process, the yield of hyaluronic acid obtained reaches 6.0g/L, and the molecular weight reaches 2.00×10 ⁶ Da.

Comparative example

Using Streptococcus zooepidemicus H23 as the starting strain, hyaluronic acid is produced by liquid fermentation. The fermentation medium uses 8.0-12.0g/L yeast powder and 8.0-12.0g/L beef extract as the compound nitrogen source and fermentation medium The other components were the same as those in Example 1, the stirring speed was about 650rpm during the fermentation process, the yield of hyaluronic acid was about 3.8g/L, and the molecular weight was about 1.21×10 ⁶ Da, both of which were lower than in Example 1.

Claims (2)

1. method that improves fermentative Production hyaluronic acid volume of production and molecular weight, it is characterized in that adopting streptococcus zooepidemicus (Streptococcus zooepidemicus) H23 as starting strain, this bacterial classification is open at " chemical industry journal " the 54th volume the 3rd phase 350-356 page or leaf, produce hyaluronic acid through liquid fermenting, it is that single nitrogenous source, fermenting process employing mixing speed are 180～220rpm that fermention medium is selected 8.0～12.0g/L yeast powder for use, to improve hyaluronic output and molecular weight.

2. method according to claim 1 is characterized in that fermention medium consists of: glucose 65.8g/L, yeast powder 8.0～12.0g/L, Na ₂HPO ₄12H ₂O 6.2g/L, K ₂SO ₄1.3g/L, MgSO ₄7H ₂O2.0g/L, FeSO ₄7H ₂O 0.005g/L; Processing condition are: mixing speed 200rpm, 37 ℃ of temperature, air flow 1.2vvm.