Method for raising n-propanol tolerance of kitasamycin producing strain

A technology of guitarmycin and production bacteria, applied in the field of fermentation, can solve the problems of reducing the synthesis of antibiotics, unfavorable synthesis of antibiotics, insufficient precursors, etc., and achieves the effects of improving tolerance, stabilizing fermentation production level, and improving fermentation production level.

Inactive Publication Date: 2016-02-24
SHANDONG FANGMING PHARMACEUTICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Thereby adding n-propanol as the precursor substance of biosynthesis can increase the output of kitasamycin, but adding n-propanol in the early stage of fermentation will interfere with the growth of mycelium, thereby reducing the synthesis of antibiotics. The exogenous precursor is in the fermentation stage. The residual n-propanol in the solution will poison the production bacteria and is not conducive to the synthesis of antibiotics
But not enough precursors

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Mutagenesis and screening: Prepare the slant strains from the kitasomycin-producing bacteria and culture them at a temperature of 28°C and a humidity of 30% to 50% for 9 days; add 0.85% normal saline to the cultivated slant strains under sterile conditions Wash 40ml of spores, pour the washed spores into a sterile Erlenmeyer flask with glass beads, then shake on a rotary shaker flask at 220r / min on a shaker for 5 minutes to activate and disperse the spores, and then use 0.85 % normal saline diluted to 10×10 spores 5 1 / ml, 1.5% n-propanol was added in the spore solution, and divided into two parts.

[0027] The slant medium is the same as the plate medium, the formula is: 20g starch, 1g magnesium sulfate, 1g potassium dihydrogen phosphate, 6g yeast extract, add purified water to 1000ml, adjust the pH value to 7.2, and then agar according to the ratio of 2.4% Divide into Erlenmeyer flasks.

[0028] Control part: take a prepared spore liquid coating plate medium, add 0.2...

Embodiment 2

[0035] The screened production bacteria with tolerance to n-propanol were inserted into the seed medium and cultured for 25 hours, inserted into the fermentation medium containing 1.8% n-propanol, cultured in a shaker flask for 80 hours, and placed in the bottle to measure the potency , The average content of kitasamycin is 9600u / ml.

[0036] In the unmutated kitasomycin-producing bacteria, the kitasamycin content only reaches 7500u / ml.

Embodiment 3

[0038] Insert the screened production bacteria with tolerance to n-propanol into 100L fermenter culture medium containing 1.8% n-propanol, culture for 70 hours, put the tank to measure the potency, and the average content of kitasamycin reaches 13000u / ml .

[0039] The results indicated that the kitasamycin-producing strains bred by this method could stably improve the fermentation production level.

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Abstract

The invention discloses a method for raising n-propanol tolerance of kitasamycin producing strain. The method comprises the following steps: using the kitasamycin producing strain as an original strain, adding 1.5% of n-propanol into a spore suspension, performing microwave mutagenesis, and performing preliminary screening and secondary screening to obtain a strain with strong n-propanol tolerance. The method has advantages of simple equipment, convenience in operation and slight radiation damage. The method is safe and can be adopted to stably raise the level of fermentation production.

Description

technical field [0001] The invention relates to a method for improving the tolerance of kitasamycin-producing bacteria to n-propanol, and belongs to the technical field of fermentation. Background technique [0002] Kitamycin is a macrolide antibiotic with broad-spectrum antibacterial activity. The antibacterial activity against most Gram-positive bacteria is slightly poor, and some erythromycin-resistant Staphylococcus aureus is still sensitive to kitasamycin. [0003] Kitamycin is also quite active against Diphtheria, Tetani, Neisseria gonorrhoeae, Pertussis, Rickettsia and Chlamydia trachomatis. [0004] Kitamycin can be taken orally or injected. The advantage of clinical application is that it is still effective against most erythromycin-resistant or penicillin-resistant Staphylococcus aureus, and it can also be used for skin and soft tissue infections, respiratory tract infections, streptococcal angina, scarlet fever, diphtheria, and Legionella caused by Gram-positive...

Claims

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Application Information

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IPC IPC(8): C12N15/01C12N13/00
Inventor 沈玲霞余洪智于新令
Owner SHANDONG FANGMING PHARMACEUTICAL CO LTD
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