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Streptomyces rubiginosohelvolus capable of producing actinomycin D through fermentation and applications thereof

A technology of Streptomyces celuteus and Streptomyces cereus, applied in the field of fermentative production of actinomycin D, can solve the problems of unsatisfactory performance of fermentation strains and achieve stable potency, Improved yield and good stability

Active Publication Date: 2021-04-20
FUJIAN INST OF MICROBIOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] For this reason, the technical problem to be solved by the present invention is to provide a kind of Streptomyces cerevisiae mutagenesis strain that produces actinomycin D by fermentation, to solve the problem of unsatisfactory performance of the fermentation strain of actinomycin D in the prior art ;

Method used

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  • Streptomyces rubiginosohelvolus capable of producing actinomycin D through fermentation and applications thereof
  • Streptomyces rubiginosohelvolus capable of producing actinomycin D through fermentation and applications thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0030] Embodiment 1 sets out the isolation of bacterial strain FIM-Z19

[0031] Soil was collected in a vegetable garden at the foot of Mount Tai in Jinan City, Shandong Province. Specifically, the floating soil about 10cm above the surface was removed with a sampling shovel, and 10g-25g of soil samples at 10cm were collected; 2g of soil samples were weighed and 10mL of sterile saline was added. Set aside for 30 minutes, take the supernatant, that is, the original solution, and carry out gradient dilution with sterile saline, and the selected dilution is 10 -2 、10 -3 、10 -4 and 10 -5 suspension.

[0032] Take 0.1mL of the original solution and the selected suspension and spread them on the separation agar medium plate prepared by adding 50mg / L potassium dichromate water, repeat 3 parallel plates for each sample, and then place the coated plate on Cultivate at 28°C, and observe the morphological characteristics such as colony appearance, size, color, edge shape, and surface...

Embodiment 2

[0034] The acquisition of embodiment 2 mutagenesis strain FIM-Z19-12

[0035]Take the above-screened strain FIM-Z19 and transfer it to the slant medium, and culture it in a constant temperature incubator for 8-12 days at a temperature of 28°C; then wash the spores on the slant medium with physiological saline, glass The beads were broken up and filtered through gauze to make 10 6 individual / mL spore suspension.

[0036] Use a pipette gun to draw 10 μL of the spore suspension prepared above onto a circular iron sheet with a diameter of 1 cm, and place it in a room-temperature plasma induction chamber with helium as the working gas, a power supply of 110 W, and a working gas flow rate of 10 L / min. In the variable system, and the treatment distance is 2mm, respectively treat 5s, 10s, 15s, 30s, 45s, 60s, 75s, 90s, carry out gradient dilution and smear the spore suspension after treatment on a plate, and make a lethality curve (such as figure 1 shown). It can be seen from the fi...

Embodiment 3

[0046] Identification of embodiment 3 mutagenic strain FIM-Z19-12

[0047] Identification of Physiological and Biochemical Characteristics

[0048] The obtained strain FIM-Z19-12 was streaked on the separation medium plate and inserted into a cover glass, cultured at 30°C for 7-20 days, and the morphological characteristics of the single colony and its bacteria were observed with an optical microscope, transmission and scanning electron microscope. Silk.

[0049] The main morphological and physiological and biochemical characteristics of the strain FIM-Z19 are as follows: plate colonies are round, with central protrusions, wrinkled surfaces, aerial hyphae are white or yellowish, base hyphae are light yellow, and produce soluble yellow or reddish-brown Glucose, fructose, rhamnose, arabinose, mannitol can be used, sucrose, raffinose, inositol can not be used . This strain is a highly aerobic bacterium, the optimum growth temperature is 28-32°C, the optimum growth pH is 7.0-8....

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Abstract

The invention belongs to the field of microorganisms, specifically relates to streptomyces rubiginosohelvolus capable of producing actinomycin D through fermentation, and further discloses applications of the streptomyces rubiginosohelvolus in producing the actinomycin D through fermentation. Through an atmospheric and room temperature plasma mutagenesis technology, the streptomyces rubiginosohelvolus FIM-Z19-12 capable of highly yielding the actinomycin D can be obtained through screening; the strain can efficiently ferment the actinomycin D; in fermentation experiments, the potency of the streptomyces rubiginosohelvolus FIM-Z19-12 producing the actinomycin D through fermentation can reach up to 1383 mg / L, so that the yield of the actinomycin D can be greatly increased; and therefore, the streptomyces rubiginosohelvolus can be applied to industrial fermentation production.

Description

technical field [0001] The invention belongs to the field of microorganisms, in particular to a Streptomyces cerevisiae strain producing actinomycin D by fermentation, and further discloses the application of the actinomycin D production by fermentation. Background technique [0002] Actinomycin D (ActinomycinD, AMD, dactinomycin) was first isolated from the actinomycete S.melanochromogenes No.1779 or S.Parvullus, which belongs to the actinomycin family. Produced by microorganisms of the genus Streptomyces. Actinomycin D is a chromopeptidolactone antibiotic, which is one of the classic anti-tumor drugs. It has good anti-tumor efficacy, and it can be used in combination with other anti-tumor drugs and radiation therapy to achieve good results. The anti-cancer effect of actinomycin D may be closely related to its chromophore structure, and the cyclic peptide part may act as a carrier. Its unique cyclic peptide chemical structure, anti-tumor mechanism of inhibiting RNA synthes...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P21/04C12R1/465
Inventor 张祝兰杨煌建严凌斌连云阳陈洲琴程贤
Owner FUJIAN INST OF MICROBIOLOGY
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