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Compound mutation of immunosuppressive agent microbial strains

An immunosuppressant and compound mutagenesis technology, applied in the field of selective breeding and mutagenesis, can solve the problems of low fermentation activity and inability to meet the requirements of industrial production, etc.

Inactive Publication Date: 2012-09-19
CHONGQING FAGEN BIOMEDICAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Strains directly isolated from nature generally have relatively low fermentation activity and cannot meet the requirements of industrial production

Method used

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  • Compound mutation of immunosuppressive agent microbial strains
  • Compound mutation of immunosuppressive agent microbial strains
  • Compound mutation of immunosuppressive agent microbial strains

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0026] Example 1. Compound mutagenesis of Streptomyces hygroscopicus

[0027] Cultivation of Streptomyces hygroscopicus:

[0028] Connect Streptomyces hygroscopicus into the shake flask culture medium (the composition of the shake flask medium (g / L): soluble starch 10, peptone 6, yeast extract 2, potassium dihydrogen phosphate 1, glycerol 2, adjust the pH to 7.0), 250ml shake flask with 55ml of culture solution, 28°C, 200rpm, culture for 2 days.

[0029] UV mutagenesis of Streptomyces hygroscopicus:

[0030] Take 5mL of the above-cultured Streptomyces hygroscopicus suspension and transfer it into a sterile petri dish with a diameter of 90mm, put it in a sterile magnetic stirring rod, and place it on a magnetic stirrer with a power of 15W and a wavelength of 260nm at 30cm under the UV lamp. Open the lid of the dish Irradiate while stirring, with doses of 10, 15, 20, 25, and 30 min. Collect the bacterial suspension.

[0031] Nitrosoguanidine (NTG) mutagenesis of Streptomyces hygroscopi...

example 2

[0038] Example 2: Compound mutagenesis of Streptomyces tsukuba

[0039] Cultivation of Streptomyces tsukuba:

[0040] Scrape an appropriate amount of spores on the seed plate to inoculate the shake flask medium (the shake flask medium composition: dextrin 1.5%, corn steep liquor 1.5%, glycerol 0.5%, anhydrous glucose 1.0%, (NH 4 ) 2 SO 4 0.3%, CaCO 3 0.2%, pH 7.0), 28°C, 200 rpm shaking culture for 3 days.

[0041] UV mutagenesis of Streptomyces tsukuba:

[0042] Take the above cultured Streptomyces tsukuba suspension and transfer it into a sterile petri dish with a diameter of 90mm, put it in a sterile magnetic stirring rod, and place it on a magnetic stirrer with a power of 15W and a wavelength of 260nm at 30cm under the UV lamp. Open the lid of the dish Irradiate while stirring, with doses of 10, 15, 20, 25, and 30 min. Collect the bacterial suspension.

[0043] Nitrosoguanidine (NTG) mutagenesis of Streptomyces tsukuba:

[0044] Prepare a spore suspension with 0.1 mol / L, pH 6.0 p...

example 3

[0050] Example 3: Compound mutagenesis of Fusarium solani

[0051] Cultivation of Fusarium Solanum:

[0052] Put Fusarium solani into the shake flask medium (the composition of shake flask medium (g / L): glucose 10, fructose 10, corn syrup 8, soybean meal 8, pH 5.6), and the amount of liquid in the shake flask is 100 ml / 500ml shake flask, 26℃, 250rpm, culture for about 55h.

[0053] NTG mutagenesis of Fusarium Solanum:

[0054] Prepare a spore suspension with 0.1 mol / L, pH 6.0 phosphate buffer to make the concentration 10 8 The suspension was treated with NTG at concentrations of 0.5, 1, 1.5, 2, 3, and 4 mg / mL, shaking at 28°C for 30 min, and washing the spores by centrifugation with normal saline three times, and collecting the bacterial suspension.

[0055] Ion implantation mutagenesis of Fusarium Solanum:

[0056] The energy used is 5, 10, 15, 20, 25 kev, and the doses are 0.5, 1, 1.5, 2 (10 17 / cm 2 ) N + Inject into the bacteria body, the pulse time of each pulse is 5, 10, 15, 20s,...

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Abstract

The invention relates to a physical and chemical compound mutation method and screening of immunosuppressive agent microbial strains including comprises streptomyces hygroscopicus, streptomyces tsukubaensis, fusarium solani and penicillium brevicompactum. The yield of the immunosuppressive agent microbial strains is respectively improved by 35-60% by adopting compound mutation and compression mutation.

Description

Technical field [0001] The invention relates to the mutagenesis technology of microbial strains, in particular to the breeding and mutagenesis technology of four strains of Streptomyces hygroscopicus, Streptomyces tsukuba, Fusarium solani and Penicillium brevifolia. Background technique [0002] Immunosuppressants are drugs that can suppress the immune response. Selective immunosuppressants are specific to lymphocytes, but have little effect on red blood cell production or have very low toxicity, and do not damage the host's defense mechanism against bacterial and fungal infections. It has been widely used in the treatment of organ transplantation against rejection and autoimmune diseases. Research on immunosuppressants abroad has a long history. Since the first successful kidney transplantation between identical twins in 1954, medical scientists have also made many major breakthroughs in organ transplantation. In recent years, domestic and foreign Scholars are also actively wor...

Claims

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Application Information

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IPC IPC(8): C12N13/00C12N15/01C12R1/55C12R1/465C12R1/77C12R1/81
Inventor 范开张益马鲁南薛国希
Owner CHONGQING FAGEN BIOMEDICAL
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