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Method for producing demethylated tetracycline

A technology of tetracycline and chlortetracycline, applied in the field of microbial fermentation, can solve the problems of few research reports on nortetracycline, and achieve the effect of simplifying synthesis steps, reducing production costs, and improving fermentation units

Pending Publication Date: 2020-10-20
HEBEI SHENGXUE DACHENG PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In the current research, there are many studies on the synthesis of OTC, CTC, TC, and DMCTC by fermentation, and all of them have been industrialized, but there are few research reports on nortetracycline (DMTC).

Method used

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  • Method for producing demethylated tetracycline
  • Method for producing demethylated tetracycline
  • Method for producing demethylated tetracycline

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Taking the demeclotetracycline production strain Streptomyces aureus S.aureofaciens DC087 as the experimental strain, firstly carry out the demeclotetracycline production process control experiment, the culture medium used is:

[0023] 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%.

[0024] 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.

[0025] 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-amylas...

Embodiment 2

[0037]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. Therefore, the present invention optimizes the medium composition and process conditions.

[0038] Streptomyces aureus S.aureofaciens DC087 was used as the experimental strain, and the process conditions were changed for the production of nortetracycline. The medium ...

Embodiment 3

[0045] Streptomyces aureus S.aureofaciens DC087 was used as the experimental strain, and the process conditions were changed for the production of nortetracycline. The medium used was:

[0046] The composition and content of the slant culture medium are: the same as in Example 2.

[0047] The composition and content of seed bottle culture medium are: with embodiment 2.

[0048] The composition and content of the fermentation medium are: starch 9%, dextrin 2%, high-temperature soybean powder 3%, soybean protein powder 1%, calcium carbonate 1%, peptone 1%, ammonium sulfate 0.1%, L-lysine 0.8%, sodium bromide 0.3%, potassium dihydrogen phosphate 0.005%, corn steep liquor 0.3%, soybean oil (V / V) 1%, defoamer 0.01%, α-amylase (W / W of starch dosage) 0.05% , 2-Mercaptobenzothiazole 0.0015%. The pH was adjusted to 5.2-5.5 with sulfuric acid. 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 ...

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Abstract

The invention belongs to the technical field of microbial fermentation, and particularly relates to a method for producing demethylated tetracycline. The method comprises the following steps: inoculating a demethylated aureomycin production strain into a slant culture medium, and culturing for 10-15 days under the conditions of 25-30 DEG C and 20-60% humidity; after maturation, inoculating to a seed bottle culture medium, and carrying out shake culture for 36-50 hours at a temperature of 25-30 DEG C and humidity of 20%-60% and at a speed of 220 rpm; and after a seed liquid is mature, inoculating the seed liquid into a fermentation culture medium according to the inoculum size of 5%-10%, carrying out shaking culture for 5-8 days under the same condition, and finishing fermentation so as toobtain demethylated tetracycline fermentation liquid. According to the method, the fermentation unit of the demethylated tetracycline is improved, the industrial production level is achieved, and themethod has important significance and application value for simplifying synthesis steps, reducing dangerous factors and reducing the production cost in the production process of high-added-value products such as minocycline and tigecycline.

Description

technical field [0001] The invention belongs to the technical field of microbial fermentation, and in particular relates to a method for producing nortetracycline. Background technique [0002] Tetracycline antibiotics (Tetracyclines) are a class of broad-spectrum antibiotics with a core structure of Naphthacene ( figure 1 ), including oxytetracycline (OTC), chlortetracycline (CTC), tetracycline (TC), demeclocycline (DMCTC), demethyltetracycline (DMTC), doxycycline, minocycline, tigecycline (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] With the emergence of drug-resistant strains and the urgent need for broad-spectrum antibiotics, researchers have de...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P29/00C12N1/20C12R1/49
CPCC12P29/00C12N1/20
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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