Method for increasing yield of demethyltetracycline

A technology of tetracycline and demeclocycline, applied in the field of microbial fermentation, can solve the problems of few research reports on nortetracycline, achieve important application value and reduce production costs

Pending Publication Date: 2020-10-13
HEBEI SHENGXUE DACHENG PHARMA
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, little research has been

Method used

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  • Method for increasing yield of demethyltetracycline
  • Method for increasing yield of demethyltetracycline
  • Method for increasing yield of demethyltetracycline

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Using Streptomyces aureus S.aureofaciens DC087 as the experimental strain, firstly carry out the control experiment of demeclocycline production process, the medium used is:

[0026] The composition and content of the slant medium are: bran 3.0%, dipotassium hydrogen phosphate 0.01%, diamine hydrogen phosphate 0.015%, magnesium sulfate 0.005%, agar 2.5%.

[0027] The composition and content of the seed bottle culture medium are: 3.0% starch, 3.0% high-temperature soybean powder, 1.0% calcium carbonate, 0.2% yeast powder, 0.5% peptone, 0.2% ammonium sulfate, 0.15% potassium dihydrogen phosphate, soybean oil (V / V) 1%, pH natural.

[0028] The composition and content of the fermentation medium are: 8% starch, 3% high-temperature soybean powder, 1% calcium carbonate, 0.2% ammonium sulfate, 0.8% L-lysine hydrochloride, 0.3% sodium chloride, and 0.3% corn steep liquor %, soybean oil (V / V) 1%, defoamer 0.01-0.05%, α-amylase (W / W of starch dosage) 0.05%, pH natural.

[0029]...

Embodiment 2

[0040] It can be seen from Example 1 that the main component of the fermentation broth of Streptomyces aureus S.aureofaciens DC087 is demeclocycline, and nortetracycline is only one of its impurities, and the yield is very low. The structural difference between nortetracycline and nortetracycline is that the 7-position lacks a chlorine atom, so it is speculated that the synthesis process is to synthesize nortetracycline first, and then add chlorine to the 7-position to synthesize nortetracycline. Therefore, if the chloride ion in the medium is removed, or the reaction of adding chlorine is inhibited during the synthesis process, the synthesis of nortetracycline may be hindered, thereby accumulating nortetracycline. Tetracycline production. For this reason, the present invention firstly optimizes the technological conditions, then carries out strain selection and further optimizes the technological process.

[0041] Streptomyces aureus S.aureofaciens DC087 was used as the expe...

Embodiment 3

[0048] Using Streptomyces aureus S.aureofaciens DC087 as the experimental strain, according to figure 2 The step is to select and breed a nortetracycline high-yield strain through mutagenesis screening.

[0049] Take the S.aureofaciens DC087 sand tube, inoculate it on the slope of the test tube under sterile conditions, and cultivate it for 14-15 days at 28°C and 20%-60% humidity; after the slope matures, scrape off the spores and inoculate them into the mycelial medium. The ingredients and their contents are: 0.5% glucose, 0.8% maltose, 0.4% peptone, 0.4% yeast powder, 0.05% magnesium sulfate, 0.2% potassium dihydrogen phosphate, 0.4% dipotassium hydrogen phosphate, and 0.3% sodium chloride. 28°C, 220rpm shaking culture under 20%-60% humidity conditions for 6h, the mycelia liquid was washed twice with sterile water, and subjected to atmospheric room temperature plasma (ARTP) treatment, and the ARTP treatment conditions were: high-purity helium was used as Gas source, proces...

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Abstract

The invention belongs to the technical field of microbial fermentation, and particularly relates to a method for increasing the yield of demethyltetracycline. The method comprises the following stepsof (1) carrying out slant culture; (2) carrying out mutagenesis treatment; (3) carrying out resistance screening; (4) carrying out strain primary screening; (5) carrying out strain retesting; and (6)carrying out fermentation process optimization on strains obtained by screening in the step (5) to further improve the fermentation level of the demethyltetracycline. Through strain breeding and process optimization, the demethyltetracycline can be produced by using demeclocycline production strains, and the industrial production level is achieved; and the method has important application value for simplifying the synthesis steps of minocycline, tigecycline and other high-added-value products and reducing the production cost.

Description

technical field [0001] The invention belongs to the technical field of microbial fermentation, and in particular relates to a method for increasing the yield of nortetracycline. Background technique [0002] Tetracycline antibiotics (Tetracyclines) are a class of broad-spectrum antibiotics with a tetracene (Naphthacene) core structure, including oxytetracycline (OTC), chlortetracycline (CTC), tetracycline (TC), demeclotetracycline (DMCTC) ), nortetracycline (DMTC), doxycycline, minocycline, tigecycline, etc. (Nguyen F, Starosta AL, Arenz S, Sohmen D, Donhofer A, Wilson DN (2014) Tetracycline antibiotics and resistance mechanisms. Biological chemistry 395(5):559-75). Among these antibiotics, OTC, CTC, TC, DMCTC and DMTC are natural products, OTC is produced by Streptomyces rimosus, and CTC, TC, DMCTC and DMTC are produced by Streptomyces aureofaciens or its mutants. [0003] In actual production, take demeclocycline as raw material to synthesize nortetracycline ( figure 1 ...

Claims

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Application Information

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IPC IPC(8): C12N15/01C12N13/00C12N1/20C12P29/00C12R1/485
CPCC12N15/01C12N13/00C12N1/20C12P29/00
Inventor 王雪峰韩冰闫彩洁郑万静王绘砖徐珍袁昉
Owner HEBEI SHENGXUE DACHENG PHARMA
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