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Hypocrea lixii DLEN2008010 as well as preparation method and application of active extract of fermentation liquor thereof

A technology of DLEN2008010, marine fungi, applied in the field of microorganisms, can solve problems such as the decline of patients' memory and cognitive ability, and achieve the effect of good application prospects

Active Publication Date: 2017-02-01
GUANGDONG OCEAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Moreover, the activity of acetylcholinesterase in the synaptic gap of the remaining cholinergic neurons is too high, which degrades the neurotransmitter acetylcholine and cannot play a role in signal transmission between neurons, and the memory and cognitive ability of patients declines.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Example 1 Isolation and identification of the marine fungus Hypocrea nigricans DLEN2008010

[0061] 1. Isolation method of strain

[0062] (1) Sample: intertidal luxuriant membranous sponge in the sea area of ​​Dalian, China.

[0063] (2) Separation method

[0064] Rinse the above-mentioned fresh sponge sample twice with seawater after autoclaving at 121°C for 20 min, transfer it into a 200 mL Erlenmeyer flask, add glass beads and 50 mL of sterile seawater after autoclaving at 121°C for 20 min, and Shake at 60 rpm for 10 min, discard the supernatant, and rinse the sponge twice with sterile seawater.

[0065] Soak the above-mentioned sponge in 50 mL of 70% ethanol aqueous solution for 10 seconds, take it out, rinse with 50 mL of sterile seawater to remove residual ethanol on the surface, blot the remaining seawater with sterile absorbent paper, and burn the sterilized blade with flame The sponges were cut into small pieces, and transplanted onto flat solid medium for ...

Embodiment 2

[0081] Example 2 Activity test of the total fermented extract of the marine fungus Hypocrea nigricans DLEN2008010

[0082] 1. Obtaining the total extract from the fermentation of H. nigeria DLEN2008010

[0083] (1) Seed culture: inoculate the bacteria on a sterile seed solid medium plate from a slant, place the culture plate in a constant temperature incubator at 28°C for upside-down culture, and the culture time is 4 to 5 days;

[0084] (2) Expansion culture: fill 1L of liquid medium into a 3L Erlenmeyer flask, seal it with a gas-permeable sealing film, and then sterilize in an autoclave at 121°C and 0.1 Mpa for 20 minutes; after cooling, inoculate the above-mentioned Step (1) After cultivating, the culture medium block of the seed plate is 40-80cm 2 The proportion of inoculation, sealing, placed in a constant temperature incubator culture, natural light, culture time is 21 to 24 days, to obtain fermentation liquid.

[0085] Wherein, the culture medium used is as follows: ...

Embodiment 3

[0098] Example 3 Extraction of Active Substances from the Fermentation Broth of the Marine Fungus Hypocrea nigricans DLEN2008010

[0099] 1. Medium

[0100] (1) Seed solid medium: 20.0g sucrose, 500mL potato juice, 20.0g sea salt, 15.0g agar, natural pH.

[0101] (2) Liquid medium: 20.0g sucrose, 500mL potato juice, 20.0g sea salt, natural pH.

[0102] Among them, potato juice: take fresh potatoes, peeled and cut into 1cm 3 For every 200g of potato pieces, add 500 mL of distilled water and boil for 20 minutes, filter through double-layered gauze cloth, and dilute to 500 mL with distilled water.

[0103] 2. Extraction method

[0104] (1) Seed culture: inoculate the bacteria on a sterile seed solid medium plate from a slant, place the culture plate in a constant temperature incubator at 28°C for upside-down culture, and the culture time is 4 to 5 days;

[0105] (2) Expansion culture: fill 1L of liquid medium into a 3L Erlenmeyer flask, seal it with a gas-permeable sealing fi...

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Abstract

The invention discloses hypocrea lixii DLEN2008010 as well as a preparation method and application of an active extract of fermentation liquor thereof. The hypocrea lixii DLEN2008010 is preserved in China General Microbiological Culture Collection Center (CGMCC) on August 16, 2016, and has a preservation number of CGMCC No. 12868. Active materials of standing fermentation liquor of the hypocrea lixii DLEN2008010 are extracted and purified; the total extract of the fermentation liquor of the hypocrea lixii DLEN2008010 can well resist oxidation and inhibit the activity of acetylcholin esterase; the purified active extract of the fermentation liquor of the hypocrea lixii DLEN2008010 can well inhibit the activity of the acetylcholin esterase; therefore, the active extract of the fermentation liquor of the hypocrea lixii DLEN2008010 has good application prospect in the aspects of preparation of medicines for resisting alzheimer's disease and preparation of medicines for inhibiting the activity of the acetylcholin esterase.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a preparation method and application of a marine fungus Hypocrea nigricans DLEN2008010 and an active extract of its fermented liquid. Background technique [0002] With the aging of the world's population and the increasing incidence of Alzheimer's Disease (AD), it is imminent to find new drugs against Alzheimer's disease. [0003] Currently, the widely accepted pathological theory of Alzheimer's is the cholinergic nerve damage hypothesis. The cholinergic neurons in the patient's brain died in large numbers due to the deposition of beta-amyloid protein and neurofibrillary tangles caused by the aggregation of T-proteins. Acetylcholinesterase (AChE), as one of the important molecular chaperones, is involved in the aggregation and deposition of β-amyloid protein. Moreover, the activity of acetylcholinesterase in the synaptic gap of the remaining cholinergic neurons is too h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14A61K36/06A61P25/28A61P39/06C12R1/645
CPCA61K36/06C12N1/145C12R2001/645
Inventor 张翼鲍海燕聂影影黎钊坪刘亚月刘敬珊黎燕媚许茂鑫冯妍宋采
Owner GUANGDONG OCEAN UNIVERSITY
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