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Preparation method of apramycin industrial production strain

An apramycin and strain technology, which is applied in the field of the preparation of apramycin industrialized production strains, can solve the problem of unstable apramycin fermentation level, inability to maintain a constant number of spores, fluctuation of apramycin quality, etc. problems, to achieve the effect of shortening the growth cycle, stable fermentation process and stable quality

Pending Publication Date: 2022-04-15
浙江普洛生物科技有限公司 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the limitation of preservation of fresh slant, too many spores can not be prepared at one time, and the number of spores in the seed bottle can not be kept constant each time. The quality of seeds prepared by this process is unstable, and frequent batch changes also cause apramycin fermentation. Unstable levels, resulting in fluctuations in the quality of apramycin

Method used

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  • Preparation method of apramycin industrial production strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041]a. Single colony selection: Under sterile conditions, shake the apramycin strain freezing solution evenly, dip it with an inoculation needle and draw a line on the separation plate, transfer it to a 37°C constant temperature incubator and cultivate it for 5 days. Use a sterile inoculated shovel to pick a single colony with a diameter of 3-4 mm and transfer it to the shaker flask.

[0042] b. Shaking bacteria: place the single colony shake flask selected in step a in a constant temperature culture shaker, and culture at 37°C and 200rpm for 22h;

[0043] c. Preparation of frozen seed tubes: under aseptic conditions, mix the single-colony liquid culture prepared in step b with 20% skimmed milk evenly, and use sterile pipettes to distribute into 5ml plastic tubes, 3.0ml per tube, spin Tightly cover the tube, stick a label, write the batch number, and store it in a low-temperature freezer at -80°C;

[0044] d. Slope activation: Dissolve and unseal the seed tube prepared in s...

Embodiment 2

[0050] a. Single colony selection: Under sterile conditions, shake the apramycin strain freezing solution evenly, dip it with an inoculation needle and draw a line on the separation plate, transfer it to a 37°C constant temperature incubator and cultivate it for 5 days. Use a sterile inoculated shovel to pick a single colony with a diameter of 3-4 mm and transfer it to the shaker flask.

[0051] b. Shaking bacteria: place the single colony shake flask selected in step a in a constant temperature culture shaker, and culture at 37°C and 200rpm for 24h;

[0052] c. Preparation of frozen seed tubes: under aseptic conditions, mix the single-colony liquid culture prepared in step b with 20% skimmed milk evenly, and use sterile pipettes to distribute into 5ml plastic tubes, 3.0ml per tube, spin Tightly cover the tube, stick a label, write the batch number, and store it in a low-temperature freezer at -80°C;

[0053] d. Slope activation: Dissolve and unseal the seed tube prepared in ...

Embodiment 3

[0059] a. Single colony selection: Under sterile conditions, shake the apramycin strain freezing solution evenly, dip it with an inoculation needle and draw a line on the separation plate, transfer it to a 37°C constant temperature incubator and cultivate it for 5 days. Use a sterile inoculated spatula to pick a single colony with a diameter of 3-4 mm and transfer it to the shaker flask.

[0060] b. Shaking bacteria: place the single colony shake flask selected in step a in a constant temperature culture shaker, and culture at 37°C and 200rpm for 24h;

[0061] c. Preparation of frozen seed tubes: under aseptic conditions, mix the single colony liquid culture prepared in step b with 10% skimmed milk evenly, and use sterile pipettes to distribute into 5ml plastic tubes, 3.0ml per tube, spin Tightly cover the tube, stick a label, write the batch number, and store it in a low-temperature freezer at -80°C;

[0062] d. Slope activation: Dissolve and unseal the seed tube prepared in...

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Abstract

The invention discloses a preparation method of an apramycin industrial production strain. Comprising the following steps: selecting single colonies from a flat plate to a shake flask, culturing a seed solution in the shake flask, preparing a frozen seed tube for the seed solution, performing slant activation on the seed solution in the frozen seed tube, preserving mature slant spores in a milk frozen tube, counting viable bacteria, preparing a spores solution in a tank, and inoculating the spores in the tank. According to the preparation method of the apramycin industrial production strain, the phenomenon that the fermentation level is unstable due to frequent slope replacement in a traditional method can be avoided. The preparation method is easy and convenient to operate, the amount of spore liquid fed into the tank each time is constant, and then the stability of the seed quality of the seed tank is guaranteed.

Description

technical field [0001] The invention belongs to the preparation and cultivation method of microbial industrial fermentation strains, and in particular relates to a preparation method of apramycin industrial production strains. Background technique [0002] Apramycin, also known as apramycin, is a new aminoglycoside veterinary antibiotic produced by Streptomyces obscura. It was successfully developed by the United States in the 1980s. It is characterized by a wide antibacterial spectrum and is not easy to produce drug resistance. It can be used Infection caused by Escherichia coli, Salmonella and mycoplasma in chicken. As a drug-type feed additive, apramycin can significantly promote weight gain and improve feed conversion rate, and has been widely used in the field of livestock and poultry breeding. [0003] Apramycin is fermented and synthesized by Streptomyces obscura. At present, domestic research on the biosynthesis of apramycin by fermentation method is mainly through ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/02C12N1/04C12P19/60C12R1/465
Inventor 钱丽丽李露园吕菊琴陈瑞峰许永兴金鑫强
Owner 浙江普洛生物科技有限公司
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