Huperzine A high yield strain TCM-01

A technology of huperzine A and high-yield strains, applied in the field of microorganisms, can solve the problems of high price, complicated steps, and difficult artificial synthesis, and achieve the effect of increasing product output and saving costs

Inactive Publication Date: 2010-12-15
洪亚辉 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, the content of huperzine A in the whole plant is very small. Due to the scarcity of huperzine resources, artificial cultivation has not yet been possible. At present, the production of medicines is completely dependent on wild resources, which are expensive. sustainable development
Although there is also artif...

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] Embodiment 1, the preparation method of bacterial strain of the present invention:

[0013] Taking Penicillium chrysogenum as the starting strain, the mutant strains were screened by microwave method. Diluted spore suspension (density about 10 6 Each sample / mL) was placed in a beaker filled with ice water (the thermal effect of microwave was eliminated by ice bath method), microwave (frequency 2450MHz, rated input power 1300W, microwave output power 800W) radiation treatment time were: 0s, 20s, 50s, 80s, 110s, 130s. After the mutated spore suspension was properly diluted, 0.1 mL was drawn, spread on a PDA plate, and cultured upside down at 28°C until a single colony grew. The microwave mutagenized spore suspension was spread on the PDA medium and cultured for about 2-3 days, and the lethality rate of the mutagenized endophytic fungal spores was calculated by counting the number of single colonies. The fungal spores were fermented and cultured, and the content of hupe...

Embodiment 2

[0014] Embodiment 2, shaker fermentation production method:

[0015] Inoculate the bacterial strain of the present invention in a medium containing PDA, place it in a constant temperature shaker at 28°C and 160r / min to ferment for 8 days, filter the fermentation broth with a Buchner funnel, collect the filtrate, and then filter the filtrate with a 0.45 μm microporous membrane. Take 10 mL into scintillation vials and spin dry in a freeze-drying centrifuge. The content of huperzine A in the sample was determined by high performance liquid chromatography (HPLC), and the amount of huperzine A was 4.76 μg / ml.

Embodiment 3

[0016] Example 3, the extraction, separation and purification of the active ingredient of huperzine A: collect the fermentation broth, filter it with a Buchner funnel, collect the filtrate, and then filter the filtrate with a 0.45 μm microporous membrane, put it into a scintillation vial, pass The freeze-drying centrifuge spins and dries to obtain the Huperzine A extract.

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PUM

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Abstract

The invention relates to a huperzine A high yield strain TCM-01 of which classified nomenclature is Endophyte sp.TCM-01, conservation address is the China center for type culture collection, conservation date is April 16, 2010 and CCTCC NO is M2010086. The high yield strain uses Penicillium chrysogenum Thom as parent strain and is prepared through microwave mutagenesis-screening. The invention further optimizes the condition of fermentation culture of the strain; and under the condition of culture, the yield of huperzine A is up to 4.761mu g/ml. Under the same condition of fermentation, the yield of the product is greatly increased, and the cost can be largely saved.

Description

Technical field: [0001] The invention relates to a microorganism, in particular to a high-yielding huperzine A strain endophyte (Endophyte sp.) TCM-01. Background technique: [0002] Huperzine A (Hup A for short) is an effective monomer of lycopodium alkaloids isolated from Huperzia serrata. It is a high-efficiency, highly selective and reversible acetylcholinesterase (AChE) inhibitor, which can promote memory reproduction and enhance memory retention. In addition, it has a long acting time, easy penetration of blood and cerebrospinal fluid barriers, and oral bioavailability It has many advantages such as high degree of accuracy and less adverse reactions. In addition, huperzine A has multi-target effects. In addition to inhibiting the activity of acetylcholinesterase, it can also protect neurons through anti-oxidative stress and anti-apoptosis pathways, and can improve various experimental memory impairments. effect. In recent years, a large number of domestic clinical s...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P17/12C12R1/82
Inventor 洪亚辉刘东波程鹏刘仲华肖浪涛
Owner 洪亚辉
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