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High cephalosporin C yield bacterial breeding method

A technology of cephalosporins and high-yield strains, applied in the direction of microorganism-based methods, biochemical equipment and methods, and treatment of microorganisms with electricity/wave energy, can solve the problems of affecting fermentation yield, complicated operation, and high cost, and achieve growth advantages Obvious, overcoming the saturation effect, maintaining a high level of effect

Inactive Publication Date: 2013-12-11
SHANDONG LUKANG PHARMA
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Problems solved by technology

Ultraviolet mutagenesis, microwave mutagenesis, and chemical mutagen (acridine orange, nitrosoguanidine, etc.) mutagenesis in traditional methods have been widely used in the breeding of cephalosporin C strains, improving the fermentation level, However, with the improvement of strain resistance to traditional mutagens, the effect is getting worse and worse, which affects the breeding process of strains; the molecular biology method has strong purpose and better effect, but the method is complicated to operate and expensive High, the gene introduced in the fermentation production process is easy to lose, affecting the fermentation yield

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0031] 1) Take several pre-prepared strains of Cephalosporium acremonium cryogenic tubes from the -80°C cryogenic refrigerator, thaw quickly at 20-40°C, draw 1-2mL and inoculate into pre-sterilized 30- Put 100mL seed culture medium (3.0-5.0% corn steep liquor, 3.0-5.0% glucose, 2.0-4.0% sucrose, 0.2-1.0% calcium carbonate, pH 6-8) in a 500mL Erlenmeyer flask, place on a shaker at 20-30°C , 150-300rpm shaking culture for 2-6 days.

[0032] 2) Take 1-10 mL of the above bacterial suspension and inoculate it into a pre-sterilized fermentation medium containing 30-100 mL (corn steep liquor 3.0-5.0%, peanut cake powder 3.0-5.0%, sucrose 2.0-4.0%, calcium sulfate 0.2-5.0%) 1.0%, peptone 1.0-3.0%, ammonium sulfate 1.0-2.0%, ferrous sulfate 0.01-0.1%, calcium carbonate 0.2-0.5%, soybean oil 5-20%, pH6-8) in a 500mL Erlenmeyer flask, placed in 20 On a shaker at ~30°C, shake at 150-300 rpm for 4-10 days.

[0033] 3) Inoculate 1-10 mL of the fermentation broth from step 2 into a pre-ste...

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Abstract

The present invention discloses a high cephalosporin C yield bacterial breeding method, and relates to the field of microorganism bacterial breeding. According to the present invention, a metabolic regulation breeding method is adopted to overcome a saturation effect of bacterial on a mutagen so as to easily achieve mutation bacterial generation; a semi-continuous fermentation method is adopted to make bacterial with excellent characteristics rapidly establish growth advantages so as to increase chances of screening of excellent bacterial; and the bacterial breeding method is simple, fast and efficient.

Description

technical field [0001] The invention relates to the field of microbial strain breeding, in particular to a method for breeding cephalosporin C strains. Background technique [0002] Cephalosporin C is an important pharmaceutical intermediate, which can be synthesized into a series of less toxic 1. Cephalosporin products with good application effect, such as ceftriaxone sodium, cefixime, cefuroxime, cefamandole, etc. Now the semi-synthetic cephalosporins have been developed to the fourth generation, with a large clinical demand and a huge market space. [0003] Cephalosporin C is produced by fermentation of microorganisms such as Cephalosporium acremonium, which was first isolated in 1945, and the fermentation level increased from the initial 10ug / mL to more than 40000ug / mL, an increase of more than 4000 times, which is inseparable from the bacteria The hard work of the breeders. The breeding methods of cephalosporin C strains include traditional methods and molecular biol...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N13/00C12N15/01C12R1/645
Inventor 徐洪利左良成王文笙宋爱刚赵斐田晓梅戴晓艳
Owner SHANDONG LUKANG PHARMA
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