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Huperzia serrata endophytic fungus shiraia sp. strain for producing huperzine A

A technology of huperzine A and endophytic fungi, which is applied in the field of bioengineering, can solve the problems of low yield and inability to apply industrialized production, and achieve the effect of fast growth and great application value

Inactive Publication Date: 2011-01-12
朱笃
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the yield of huperzine A in the currently reported strains is low. For example, Li Wankui et al. optimized the fermentation conditions for the production of huperzine A by the endophytic fungus 2F09P03B of the genus Acremonium serrata, and the highest yield was only 8.32 μg / L ( "Study on Fermentation Conditions of Huperzine A Produced by Endophytic Fungus 2F09P03B of Huperz serrata", China Medical Biotechnology, 2007, 2(4): 254-259), cannot be applied to industrial production

Method used

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  • Huperzia serrata endophytic fungus shiraia sp. strain for producing huperzine A
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  • Huperzia serrata endophytic fungus shiraia sp. strain for producing huperzine A

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Effect test

Embodiment 1

[0037] Embodiment 1: preparation or screening of bamboo yellow bacterial strain of the present invention

[0038](1) Collect the complete strain of Hupercia serrata, put it in a sealed bag, store it in a 4°C incubator, and return it to the laboratory for immediate processing.

[0039] (2) Rinse the whole plant with tap water for 15-20 min, and rinse with sterile water for 3 times. Roots and stems are cut into 1-2cm segments, and the leaves are kept in their original state.

[0040] (3) Roots, stems and leaves are preliminarily sterilized by immersing in 75% ethanol for 5 minutes, then soaking in 0.1% mercuric chloride for 8 minutes, and rinsing with sterile water to sterilize the surface of the tissue.

[0041] (4) Cut off the two ends of the above-mentioned treated stem with sterilized scissors, then cut the stem with two ends longitudinally, divide it into two, and cut it into small pieces of 0.1×0.5 cm. The edge of the leaf is cut off with sterilized scissors, and the edg...

Embodiment 2

[0045] Embodiment 2: The present invention screens out the bamboo yellow bacterial strain that highly inhibits acetylcholinesterase activity

[0046] (1) Take the endophytic fungus isolated from the Hupermus serrata plant for activated culture, transfer it into the sterilized liquid PD medium under sterile conditions, and cultivate it on a shaker at 120 rpm at 28°C for 12 days;

[0047] (2) After the fermentation is completed, the mycelium is obtained by suction filtration, the mycelium is collected for cell crushing, and then placed in a Soxhlet extractor for extraction with absolute ethanol, and the extract is concentrated to 1 / 5 of the original by a rotary evaporator , add an equal volume of chloroform for extraction, combine the chloroform phases, evaporate to dryness with a rotary evaporator at 40°C, dissolve in a small amount of methanol, and use it as the sample solution to be tested.

[0048] (3) The initial screening uses ATCh as the substrate and DTNB as the chromoge...

Embodiment 3

[0051] Embodiment 3: Micromorphology and molecular biological identification of bamboo yellow bacterial strain of the present invention

[0052] Get the strain with the highest inhibition rate to carry out microscopic morphology and molecular biology method identification, the specific process is as follows:

[0053] (1) Solid plate culture method: cultured in potato dextrose PDA medium, cultured at 28±1°C for 4 days, the colony diameter was 31 mm, and the colony diameter was 65 mm at 7 days, and the entire culture dish was covered in 12 days. The hyphae are white at the beginning, filamentous intertwined, not easy to pick; the colony is dry, opaque, closely combined with the medium, the edge is uneven, radial filamentous, with white fluff on the surface, and becomes tapetal when mature, reddish, The pigment produced by the bacteria is partially secreted into the medium, making the back of the medium brownish red. The colonies were distributed in the shape of red and white co...

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Abstract

The invention discloses a huperzia serrata endophytic fungus for producing huperzine A, and belongs to the technical field of biological engineering. The fungus is obtained by separating from fern H. serrata living plant according to the endophytic fungus separation and purification technology, named as shiraia sp. slf-14 by microbial taxonomy authentication and collected in CCTCC on Nov. 6th, 2009 with the collection number of CCTCC No: M 209294. The strain can produce the huperzine A by liquid fermentation. The invention discovers the huperzine A contained in the shiraia sp. fermentation liquid for the first time, and provides a new path for seeking an active huperzine A component resource for treating senile dementia.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a fungus whose fermentation product contains huperzine A and its analogs. The fungus is of great significance for the production of huperzine A and its analogs by fermentation. Background technique [0002] Huperzine A (huperzine A), also known as fordine, is an alkaloid isolated from the fern Huperzine Huperzia genus Huperzia serrata for the first time in my country. (acetylcholine esterase, ACHE) has efficient, reversible, and highly selective inhibitory effects, and can be used for the treatment of Alzheimer's disease (AD), and has significant curative effects on myasthenia gravis, memory impairment, and vascular dementia. Huperzine A (huperzine A) is a first-in-class new drug (trade name of Huperzine) created in my country, and it is recognized as a first-in-class new drug with high therapeutic index, long duration of action, and no side effects. Compared with huperzin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12P17/12A61K31/4748A61P25/28A61P21/04C12R1/645
Inventor 朱笃张志斌曾庆桂颜明王吉祥
Owner 朱笃
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