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Penicillin v potassium high-producing bacteria and its application in the fermentation production of penicillin v potassium

A technology for penicillin V and high-yield strains, applied in fermentation, microbial-based methods, fungi, etc., can solve the problems of low fermentation unit, high production cost of penicillin V potassium, and difficulty in removal

Active Publication Date: 2021-05-14
NORTH CHINA PHARMA COMPANY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The fermentation unit of penicillin V potassium has always been low, resulting in high production costs of penicillin V potassium; penicillin V potassium is accompanied by 4-hydroxypenicillin V potassium impurities during the fermentation process, and 4-hydroxypenicillin V potassium impurities are produced during the refining process of penicillin V potassium hard to remove

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 Screening of Penicillin V Potassium High Yield Strain

[0035] (a) Preparation of Penicillium chrysogenum spore suspension: The starting strain is the self-produced strain of North China Pharmaceutical Co., Ltd. Beida Branch. Aseptically weigh 2g of millet spores and put them into a sterilized 250mL triangular flask (containing 10 glass beads), then add 30mL sterile distilled water to the flask, then place the flask on a shaker at 200r / min for 5 minutes, transfer the washed spore liquid to a sterile test tube for later use, the concentration of the spore liquid for 10 7 individual / mL.

[0036] (b) Combined mutagenesis with ultraviolet light and nitrosoguanidine: transfer 10 mL of spore liquid to a sterile glass plate, irradiate the spore liquid in the plate under a 15W ultraviolet lamp at a distance of 25 cm for 30 seconds, and then transfer the spore liquid to a sterile 100 mL Add nitrosoguanidine at a final concentration of 1.0 g / L to the Erlenmeyer flask,...

Embodiment 2

[0045] Example 2 Investigation of the Genetic Stability of Penicillin V Potassium High Yield Strains

[0046] The Penicillium chrysogenum B181126 preserved in Example 1 was inoculated on the slant culture medium prepared in Example 1, and cultured in the dark at 25±0.5° C. and relative humidity of 40%-60% for 8 days. After the slant strains matured, the It was re-inoculated in the slant medium for cultivation, and repeated four times; the five cultured strains were respectively transferred to the seed medium prepared in Example 1, at 25±0.5°C, relative humidity 40%-60 %, 220rpm under the condition of cultivating 45h, after the strains are mature, they are respectively transferred to the fermentation medium prepared in Example 1, and cultivated for 7 days under the conditions of 25±0.5°C, relative humidity 40%-60%, and 220rpm. Five groups of fermentation broths containing penicillin V potassium were obtained, and the potency of penicillin V potassium in the fermentation broth w...

Embodiment 3

[0047] Example 3 Excellent fermentation process control of penicillin V potassium high-yield strain

[0048] Preparation of rice spore medium: Weigh 4g sucrose, 20g corn steep liquor, 1g sodium chloride, 0.005g magnesium sulfate, 0.01g ferrous sulfate, dilute to 1000mL with distilled water, and then use 30% sodium hydroxide solution Adjust the pH to 6.8±0.2, pour it into a pot with 2600 grams of dry millet, stir it well, steam it in a disinfection cabinet at 100±2°C for 20 minutes, put the steamed rice on the fan outlet to cool it, rub it loose, and put the millet Divide into eggplant bottles (20g / bottle), put the eggplant bottles in a disinfection cabinet, sterilize at 121°C for 30 minutes, and cool to obtain rice spore culture medium.

[0049] Seed tank medium preparation: Weigh 300kg sucrose, 750kg corn steep liquor, 300kg ammonium sulfate, 150kg calcium carbonate, and use deep well water to set the volume to 13m 3 , then use 30% sodium hydroxide solution to adjust the pH ...

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Abstract

The present invention provides a kind of penicillin V potassium high-yielding bacterium and its application in fermentative production of penicillin V potassium, the penicillin V potassium high-yielding bacterium is Penicillium chrysogenum ( Penicillium Chrysogenum ) B181126 strain, the preservation date is October 14, 2019, the preservation number is CCTCC NO: M2019823, and the preservation unit is the China Center for Type Culture Collection (CCTCC). When Penicillium chrysogenum B181126 is used in production, with the optimal fermentation process, its fermentation unit can reach more than 80,000 U / mL, which is 36.4% higher than the starting strain, and the fermentation unit of penicillin V potassium is greatly increased. At the same time, 4 ‑ Hydroxypenicillin V potassium impurity content is less than 1.2%.

Description

technical field [0001] The invention relates to the technical field of penicillin V potassium fermentation, in particular to a penicillin V potassium high-yielding bacterium and its application in fermentative production of penicillin V potassium. Background technique [0002] Penicillin V Potassium belongs to β - Lactam antibiotics are an important raw material drug. Potassium penicillin V is not easily decomposed at low pH, so it can be processed into oral preparations and administered orally. Potassium penicillin V can also be cracked to prepare intermediates of two semi-synthetic antibiotics, 6-APA and 7-ADCA, which are used to synthesize a series of semi-synthetic penicillins and semi-synthetic cephalosporin antibiotics. [0003] Penicillin V potassium is produced by Penicillium chrysogenum ( Penicillium Chrysogenum ) are synthesized through secondary metabolism during fermentation. The fermentation unit of penicillin V potassium has always been low, resulting in hig...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P37/00C12R1/82
CPCC12N1/14C12P37/00C12N1/145C12R2001/82
Inventor 段志钢王平尹贵超王成高雨宁张军剪王淑琳王云牛宵亮李景丁宝娟李默晗杨婷马琳
Owner NORTH CHINA PHARMA COMPANY
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