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A radiation-resistant Aureobasidium pullulans and its application in the preparation of melanin

A technique of Aureobasidium pullulans and melanin, applied in the direction of microorganism-based methods, biochemical equipment and methods, fungi, etc., to achieve strong growth and reproduction capabilities, stable genetic characteristics, and improved survival rates

Inactive Publication Date: 2019-06-04
THE INST OF MICROBIOLOGY XINJIANG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Although there are many studies and reports on the use of strains to ferment and produce melanin, there are still many studies on the screening of radiation-resistant strains from the surface soil of radiation-contaminated environments, and the use of self-selection and domestication of Aureobasidium pullulans with radiation resistance. The research on the preparation of melanin by fermentation has not been reported yet

Method used

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  • A radiation-resistant Aureobasidium pullulans and its application in the preparation of melanin
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  • A radiation-resistant Aureobasidium pullulans and its application in the preparation of melanin

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Experimental program
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Effect test

Embodiment 1

[0038] Example 1: Isolation and screening of Aureobasidium pullulans MF1 CGMCC No. CGMCC 11275

[0039] The Aureobasidium pullulans used in the present invention is sampled and separated from 5cm-20cm soil on the surface of a radiation-polluted environment in Xinjiang. Take 1.0g of the collected soil samples and pack them into sterile test tubes of 15mm × 150mm. 60 10kGry dose irradiation treatment was carried out under Coγ ray, and then 5mL liquid Chardonnay medium was added to the test tube, and it was shaken and enriched at 28°C for 3d. Take 1 mL of the above enrichment solution and transfer it into a test tube containing 9 mL of sterile physiological water. After successively carrying out gradient dilution, take 0.2 mL of the dilution solution and spread it on the Chaplain medium plate, cultivate it at a constant temperature of 28 °C for 7 days, and observe the colonies. Growth, pick a single colony for purification. Some of the purified strains were preserved in lyophil...

Embodiment 2

[0043] Example 2: Isolation, screening and identification of Aureobasidium pullulans MF1 CGMCC No. CGMCC11275

[0044] 1. PCR amplification of the rDNA ITS segment of Trichoderma viride and its sequencing

[0045] The strains were inoculated into the modified Char's medium, cultured at a constant temperature of 28 °C for 4 days, and the bacteria were collected by centrifugation. Referring to Gerrits et al., the total DNA extraction method was used for PCR amplification with fungal universal primers in the D1 / D2 region of LSU rDNA, and the primers were designed. as follows:

[0046] NL1: 5′-GCATATCGGTAAGCGGAGGAAAAG-3′,

[0047] NL4: 5′-GGTCCGTGT-TT CAAGACGG-3′;

[0048] The PCR product was purified and recovered by cutting gel, dissolved in deionized water, and sequenced.

[0049] 2. Sequence alignment and phylogenetic analysis of LSU rDNA D1 / D2 regions

[0050] The LSU rDNA D1 / D2 region sequences obtained from the strains were compared with the known sequences in the GenBa...

Embodiment 3

[0051] Example 3: Extraction and preparation of crude melanin extract by Aureobasidium pullulans MF1 CGMCCNo.CGMCC11275

[0052] (1) Activation of strains: Aureobasidium pullulans MF1 CGMCC No. CGMCC11275 was inoculated into a modified modified Chapson solid medium, and cultured at 28°C for 48 hours to activate the strains.

[0053] Described improved Chase solid medium: sodium nitrate 3g, yeast extract powder 0.5g, dipotassium hydrogen phosphate 1g, magnesium sulfate (MgSO4) 4 ·7H 2 O) 0.5g, potassium chloride 0.5g, ferrous sulfate 0.01g, glucose 30g, agar 20g, distilled water 1000mL, natural pH, subpackage, sterilize.

[0054] (2) Cultivation of strains: inoculate the activated strain in step (1) in the modified Chaplain liquid medium, wherein the loading amount of the modified Chaplaincy liquid medium is 100mL, glucose 10.3g / L, yeast extract powder 2.02g / L, ammonium sulfate 2.0g / L, NaCl 0.5g / L, bacterial inoculum volume 2mL, 30°C, 180r / min shaker culture for 96h to obtai...

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Abstract

The invention discloses radiation-resisting aureobasidium pullulans and application thereof in preparing melanin. The Aureobasidium pullulans MF1 CGMCC No. CGMCC11275 is obtained by carrying out separation, screening, breeding and acclimation on surface soil of a certain radiation pollution environment in Sinkiang; the melanin is obtained on the conditions that the Czapek-Dox medium addition amount is 100 mL, the glucose addition amount is 10.3 g / L, the yeast extract powder addition amount is 2.02 g / L, the ammonium sulfate addition amount is 2.0 g / L, the NaCl addition amount is 0.5 g / L, the bacteria inoculation amount is 2 mL, and shake cultivation is carried out at 30 DEG C at the speed of 180 r / min for 96 h. The obtained melanin can effectively improve the survival rate of bacterial strains under ultraviolet radiation, and has the realistic meaning and effect.

Description

technical field [0001] The present invention relates to the technical field of microorganism application, in particular, the present invention relates to a kind of Aureobasidium pullulans and the technical field of its application in melanin fermentation. Background technique [0002] Natural melanin is a heterogeneous polyphenol-like polymer that widely exists in animals, plants and microorganisms, and is a general term for a class of high molecular weight brown-melanin. Melanin has good light absorption in both ultraviolet and visible light wavelengths, especially under ultraviolet light (maximum absorption wavelength is 210nm), it is generally soluble in alkaline solution, insoluble in acidic solution, slightly soluble in Water, insoluble in common organic solvents. Melanin has strong functions such as scavenging free radicals and anti-oxidation, which are superior to conventional antioxidant products such as SOD and vitamin E. At the same time, some soluble melanin has ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12P1/02C12R1/645
Inventor 朱静房世杰樊国全张志东孙翔楚敏王玮宋素琴顾美英唐琦勇
Owner THE INST OF MICROBIOLOGY XINJIANG ACADEMY OF AGRI SCI
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