Streptomyces rimosus mutant strain and preparation method and application thereof

A technology of Streptomyces fissures and mutant strains, applied in the field of strains for preparing tetracycline antibiotics, can solve the problems of unsatisfactory defoaming effect, adverse effects on growth and metabolism of Streptomyces fissures strains, etc., and achieves good passage stability and results. Reliable, good antifoaming effect

Active Publication Date: 2015-04-01
HEBEI SHENGXUE DACHENG PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the defoaming effect of this method is not ideal
If the concentration of defoamer is increased, although the defoaming effect can be further enhanced, high-concentration defoamer will have adverse effects on the growth and metabolism of Streptomyces crackis; especially when the amount of defoamer is greater than 0.1% (mass volume ratio Concentration), it will have a significant inhibitory effect on the strain

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Preparation of Mutant Strain of Streptomyces rimosus

[0030] a. Slope culture: The initial strain of Streptomyces rimosus is activated and inoculated into the pre-prepared bran slant medium, first cultivated at 32-35℃ and 30-40% relative humidity for 48-60 h, and then Cultivate for 20-25 h at 25-30℃ and 25-35% relative humidity.

[0031] The formula of the bran slant medium prepared in this embodiment is: 30 g of bran, 10 g of agar, and water to 1000 ml; the preparation method is a conventional method.

[0032] b. Seed culture: Transfer the strains after slant culture to the pre-prepared seed culture medium, and cultivate them with shaking at 190-200 rpm at 25-28°C for 25-27 h.

[0033] The formula of the seed culture medium prepared in this embodiment is: starch 20g, soybean flour 1g, calcium carbonate 1g, ammonium sulfate 2g, sodium chloride 3g, corn steep liquor 2g, potassium dihydrogen phosphate 0.01g, and water to 1000ml. The preparation method is a conventiona...

Embodiment 2

[0041] Example 2 Retest and stability investigation of mutant strains

[0042] Strain retest: Take the sandy soil tube of DC461 preserved in Example 1, and inoculate it on the bran slant medium prepared in Example 1. After incubating for 48-120 h at 32-40°C and 30-60% relative humidity, Cultivate for 20-40 h at 25-35℃ and relative humidity of 25-65%, dig out about 0.5×0.5 cm with an inoculating shovel 2 The bacterial layer was inoculated on the seed medium prepared in Example 1, and after shaking culture at 28.0±0.5°C and 220 rpm for 26-30 h, it was transferred to the fermentation medium A containing defoamer prepared in Example 1 , At 25-35℃, 190-240 rpm shaking culture for 72-192h, the titer of oxytetracycline in the fermentation broth was determined, and the result was 16016 U / mL. This result shows that the DC461 strain has good activity, high yield and anti-foaming agent characteristics.

[0043] Stability investigation: Take the sandy soil tube of DC461 preserved in Example 1...

Embodiment 3

[0044] Example 3 Application of DC461 in the production of oxytetracycline

[0045] Take the strain DC461 selected in Example 1 as the fermentation strain, inoculate it in the fermentation medium, and ferment for 120h at 25-28°C and 190-rpm. The total volume of the fermentation medium is 10L, including starch 1200g, soybean meal 200g, calcium carbonate 150g, ammonium sulfate 150g, sodium chloride 40g, corn steep liquor 40g, potassium dihydrogen phosphate 1g, cobalt chloride 0.1g, and amylase 1.2 g. 50g polyether defoamer, the rest is water. After the fermentation was completed, the fermentation broth was taken, centrifuged at 10000 rpm for 5 min, and the supernatant was taken, and the oxytetracycline titer was detected, and the result was 15776 U / mL. This result indicates that the strain DC461 screened in the present invention can be used for the production of oxytetracycline.

[0046] In the fermentation broth of this example, the concentration of the defoamer was increased to 0...

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PUM

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Abstract

The invention provides a streptomyces fungicidious mutant strain DC461, wherein the collection number is CGMCC (China General Microbiological Culture Collection Center) No. 5811, the collection unit is China Committee for Culture Collection of Microorganisms, and the collection date is February 27, 2012. The invention further provides a preparation method of the strain and application of the strain in terramycin production. The mutant strain DC461 provided by the invention is bearable to a high-concentration defoamer and can be used for increasing the production level of terramycin.

Description

Technical field [0001] The invention relates to a strain for preparing tetracycline antibiotics, in particular to a mutant strain of Streptomyces rimosus and its preparation method and its application in the production of oxytetracycline. Background technique [0002] Terramycin (Terramycin), also known as "diomycin" or "oxotetracycline", is the second broad-spectrum tetracycline antibiotic to be discovered. It is resistant to Gram-positive bacteria, Gram-negative bacteria, Mycoplasma, Chlamydia, Rickettsia The infection caused by the body is effective, and it can also be used to treat piriformis disease and eperythrozoon disease. Oxytetracycline is produced by the fermentation of Streptomyces rimosus. At present, when oxytetracycline is produced by fermentation, due to the selection of starting strains and the influence of oxytetracycline cell growth and metabolism, it is easy to form stable and difficult-to-eliminate foam in the fermentation broth. These bubbles not only hind...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N15/01C12N13/00C12P29/00C12R1/59
Inventor 齐巧娟赵丽红
Owner HEBEI SHENGXUE DACHENG PHARMA
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