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Penicillium ochrochloron and its application in preparation of brefeldin A

A technology of Penicillium ochracea and a preservation number is applied to Penicillium ochracea and its application field in the preparation of brefeldin A, which can solve the problems of many by-products, low fermentation level, easy degradation of bacterial species, etc., and achieves great application value Effect

Inactive Publication Date: 2015-06-17
SOUTHWEST UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In 2005, Chinese scientist Liu Wanyun also reported a Penicillium fungus SHZK-15 that produced BFA, but the yield was only 151.6mg / L fermentation broth
The above studies all provide the possibility for the industrial production of BFA, but because these strains are easy to degrade, the fermentation level is generally low, and there are many by-products, it is difficult to meet the growing commercial production requirements

Method used

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  • Penicillium ochrochloron and its application in preparation of brefeldin A
  • Penicillium ochrochloron and its application in preparation of brefeldin A
  • Penicillium ochrochloron and its application in preparation of brefeldin A

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Embodiment 1

[0030] The isolation of embodiment 1 bacterial strain

[0031] (1) Strain screening: the collected plant samples were surface-sterilized and cut into small pieces of 0.5 cm in length, and inoculated into the center of a PDA plate containing 60 μg / mL streptomycin and 100 μg / mL ampicillin. Cultivate at 28°C for 2 to 15 days, transfer a single strain to fresh PDA medium in time by using the tip mycelia picking method, and then repeat the separation of mycelia until it is a pure culture, number the obtained purified strains, and Transferred to fermentation medium (medium composition: potato 200g / L, glucose 20g / L), 28°C, 150rpm shaking culture for 15 days. The fermentation broth was centrifuged, the supernatant was collected, and extracted 3 times with 100 mL ethyl acetate, the extract phase was collected, the ethyl acetate was recovered by distillation under reduced pressure, the obtained crude extract was dissolved in methanol, and thin layer chromatography (Thin Layer Chromatogr...

Embodiment 2

[0043] Example 2 Effect of Different C Sources on the Preparation of BFA by Fermentation of Penicillium ochregreen SWUKD4.211

[0044] In order to investigate the influence of different C sources on the fermentation level of BFA in the medium, five different fermentation mediums were designed. The composition is respectively:

[0045] Medium A: potato 200.0g / L, xylose 20.0g / L, ammonium sulfate 2.0g / L, potassium dihydrogen phosphate 1.0g / L, magnesium sulfate heptahydrate 2.0g / L, calcium carbonate 5.0g / L, solvent For distilled water, pH=6;

[0046] Medium B: potato 200.0g / L, glucose 20.0g / L, ammonium sulfate 2.0g / L, potassium dihydrogen phosphate 1.0g / L, magnesium sulfate heptahydrate 2.0g / L, calcium carbonate 5.0g / L, the solvent is distilled water, pH=6;

[0047] Medium C: potato 200.0g / L, sucrose 20.0g / L, ammonium sulfate 2.0g / L, potassium dihydrogen phosphate 1.0g / L, magnesium sulfate heptahydrate 2.0g / L, calcium carbonate 5.0g / L, the solvent is distilled water, pH=6;

...

Embodiment 3

[0052] Example 3 Effects of Different Rotating Speeds on the Preparation of BFA by Fermentation of Penicillium ochregreen SWUKD4.211

[0053] In order to investigate the influence of rotating speed on the fermentation level of BFA, three different shaking table rotating speeds were designed, and the bottling volume of the fermentation medium was 100mL / 250mL triangular flask (containing 2 glass beads with a diameter of 4mm), and the composition of the fermentation medium was For: potato 200.0g / L, glucose 20.0g / L, ammonium sulfate 2.0g / L, potassium dihydrogen phosphate 1.0g / L, magnesium sulfate heptahydrate 2.0g / L, calcium carbonate 5.0g / L, solvent is distilled water, pH=6, sterilized at 121°C for 20 minutes.

[0054]The bottling volume of the fermentation medium is 100mL / 250mL Erlenmeyer flask. Under sterile conditions, use a puncher (6mm in diameter) to take a certain amount of activated strains and transfer them to the fermentation medium. , 170rpm shaker for 7 days. After ...

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Abstract

The invention belongs to the technical filed of microbes, and concretely relates to Penicillium ochrochloron and its application in the preparation of brefeldin A. A technical problem to be solved in the invention is providing a new choice for production of BFA through biological fermentation. A bacterial strain produced in the invention is Penicillium ochrochloron SWUKD4.211, and has a preservation number of CCTCC M2015022. The invention also provides a method for producing the BFA by adopting Penicillium ochrochloron fermentation. The BFA production ability of the bacterial strain obtained through the method can reach 356.0mg / L, and the method provides a new choice for the production of the BFA, provides a precisou strain for biological pharmacy, and has great application values.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to Penicillium ochrogreen and its application in preparing brefeldin A. Background technique [0002] Brefeldin A (BFA for short) is a naturally occurring macrolide antibiotic with the chemical name [(1R,2E,6S,10E,11as,14aR)-1,6,7,8 ,9,11a,12,13,14,14a-Decahydro-1,13-dihydroxy-6-methyl-4H-cyclopent[f]oxacyclotridecin-4-one], the molecular formula is C 16 h 24 o 4 , the molecular weight is 280, and the structural formula is as follows: [0003] [0004] Singleton was the first to isolate BFA from Penicillium decumben in 1958, but it was named decumbin. According to their study, BFA has also been shown to be toxic to rats and goldfish. In 1962, Betina et al. isolated the compound cyanein from Penicillium cyaneum. They found that the compound inhibited the growth of Candida albicans, Saccharomyces cerevisiae and human cervical cancer cells. Among them, the averag...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P17/08C12R1/80
CPCC12P17/08C12N1/145C12R2001/80
Inventor 黄谦董锦艳毛洪强
Owner SOUTHWEST UNIVERSITY
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