Saccharopolyspora erythraea and application thereof

A technology for Saccharopolyspora rubrum and strains, which are applied to Saccharopolyspora rubrum and its application field in fermentation production of erythromycin, and achieve the effects of good genetic stability, improved efficiency and low cost

Active Publication Date: 2013-02-06
北京燕化佰佳信作物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Later, after years of strain selection and process optimization, the fermentation unit gradually increased. At present, the domestic average fermentation unit can reach 8000μg / mL, but there is still a big gap with the current international 18000μg / mL. The lower fermentation unit is still It is an important factor restricting the development of erythromycin fermentation industry in my country

Method used

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  • Saccharopolyspora erythraea and application thereof
  • Saccharopolyspora erythraea and application thereof
  • Saccharopolyspora erythraea and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Isolation and identification of embodiment 1 bacterial strain

[0027] 1. Isolation of strains

[0028] Soil samples were randomly collected from Ning'an City, Heilongjiang Province. Gradiently dilute the frozen preserved soil samples, apply 2-3 pieces of double-disc culture medium for each dilution, and incubate at 34°C for 2-6 weeks. Dry the double dish; after that, observe the growth of the double dish with a phase contrast microscope every 2-3 days, and transfer the grown microbial colony to a new double dish (if it is found that the tiny colony no longer grows, pick the colony into the liquid 10% LB The medium was cultured statically for about 2 weeks. After the mycelia grew in the liquid, the bacteria had recovered and then double-disc streaking was performed), and the morphology was observed.

[0029] 2. Identification of strains

[0030] Saccharopolyspora rubrum is an aerobic Gram-positive actinomycete. The aerial mycelium is pinkish yellow, and the vegetativ...

Embodiment 2

[0038]Embodiment 2 fermentation produces erythromycin

[0039] The preparation of culture medium

[0040] 1. Medium formula: corn starch 12g / L, corn steep liquor 6g / L, magnesium sulfate heptahydrate 1g / L, calcium carbonate 3g / L, agar 22g / L, the balance is water, and sodium hydroxide solution before sterilization Adjust the pH to 7.2.

[0041] 2. Preparation method: first add 700mL of distilled water to a 1L beaker, heat it on an electric heating plate with an asbestos net, weigh 12g of cornstarch and dissolve it in 100mL of water, and when the distilled water is heated to about 70-80°C, dissolve the dissolved Slowly pour the starch solution into distilled water and stir it with a glass rod at a constant speed to prevent the starch from hardening due to rapid heating. After the starch solution is clarified, immerse the beaker in cold water to cool. Weigh the corresponding amount of corn steep liquor and magnesium sulfate heptahydrate into a 50mL beaker, dissolve them in 30mL ...

Embodiment 3

[0055] Example 3 Extraction and yield detection of erythromycin.

[0056] 1. Chemical detection.

[0057] 1) Test solution and preparation:

[0058] A. 0.35% Potassium Carbonate: Dissolve 35g of Potassium Carbonate in 10000mL of water and shake well.

[0059] B. Butyl acetate: Transparent and clear industrial butyl ester.

[0060] C.0.1mol / L hydrochloric acid: Dilute 90mL hydrochloric acid to 10000mL with water, shake well.

[0061] D.8mol / L sulfuric acid: slowly pour 450mL of sulfuric acid into 550mL of water, stir well, and calibrate after cooling.

[0062] 2) Drawing of standard curve:

[0063] A. Accurately weigh an appropriate amount of erythromycin standard substance, add a small amount of ethanol to dissolve, and dilute to 100mg / mL with distilled water.

[0064] B. Accurately measure 1mL, 2mL, 3mL, 4mL, and 5mL in a test tube, dilute to 5mL with water, add 5mL of 8mol / L sulfuric acid, shake well, and keep warm in a 50°C water bath for 30 minutes.

[0065] C. After...

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Abstract

The invention provides a new Saccharopolyspora erythraea (Saccharopolyspora erythraea) strain BJX001 with a preservation number of CGMCC No.4767. The invention further provides application to obtain erythrocin through the Saccharopolyspora erythraea. The experimental results show that a fermentation method of the strain, provided by the invention, has strong capacity of producing a component A of the erythrocin, a chemical titer for producing the erythrocin is up to 10000 mmg / mL of fermentation liquid, and after the strain is continuously inherited for five generations, the capacity of producing the effective component A of the erythrocin is kept at the original level, so that the heredity stability is good, the efficiency of production of the erythrocin is improved, and the method is simple and low in cost, and is suitable for popularization and application in production of the erythrocin.

Description

technical field [0001] The invention relates to the field of biological fermentation, in particular to a red saccharopolyspora and its application in fermentative production of erythromycin. Background technique [0002] Erythromycin is a macrolide antibiotic with an antibacterial spectrum similar to that of penicillin. It has antibacterial activity especially against Gram-positive bacteria, acid-fast bacilli, Rickettsia and large viruses. It is the treatment of drug-resistant golden yellow Drug of choice for diseases caused by staphylococcal infections and hemolytic streptococcal infections. Its mechanism of action is mainly to combine with the 50S subunit of ribonucleosomes, inhibit peptidyltransferase, affect the translocation process of ribonucleosomes, hinder the growth of peptide chains, inhibit the synthesis of bacterial proteins, and thus play an antibacterial role . During the fermentation process, Saccharopolyspora rubrum will produce three components, A, B, and ...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P19/62C12R1/645
Inventor 徐兵周贤龙王伟汪爱群
Owner 北京燕化佰佳信作物科技有限公司
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