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Application of endophytic fungus in transforming andrographolide diterpenoids

A new andrographolide, strain technology, applied in the field of microorganisms, can solve problems such as affecting the development of preparations

Inactive Publication Date: 2017-06-27
SHANGHAI UNIV OF T C M
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because it is insoluble in water, it can only be administered orally, which affects the development of preparations. Therefore, it is necessary to carry out structural modification of neoandrographolide to discover andrographolide derivatives with novel structures and to study their biological activities. and application pathways, providing valuable lead compounds for the research of new andrographis paniculata

Method used

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  • Application of endophytic fungus in transforming andrographolide diterpenoids
  • Application of endophytic fungus in transforming andrographolide diterpenoids
  • Application of endophytic fungus in transforming andrographolide diterpenoids

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Isolation, purification and identification of Andrographis paniculata endophyte strain LM19J01

[0031]Take the root section of the fresh plant Andrographis paniculata, rinse it, immerse it in 70% alcohol for disinfection for 4 minutes, rinse it with sterile water for 5 times, soak it in 1% sodium hypochlorite solution for 5 minutes, rinse it with sterile water for 5 times, and then plant the root section Place the explants on the PDA medium for 2 weeks in an incubator at 28°C, and then take the explants without fungal contamination and sterilize them with 1% sodium hypochlorite solution for 15 minutes, cut them and inoculate them in the PDA medium for cultivation. Use an inoculation needle to pick and inoculate the hyphae growing from the edge of the sample on a PDA medium plate, culture at a constant temperature of 28°C, and after new colonies grow, pick the tip mycelium and transfer it to a PDA medium plate to repeat purification More than 5 times, and then...

Embodiment 2

[0040] Example 2 Utilize Andrographis paniculata endophytic fungus strain LM19J01 to transform neoandrographolide

[0041] PDA solid medium: the method is to wash and peel the potatoes first, then weigh 200g of potatoes and cut them into small pieces, add water and boil them (boil for 20-30 minutes, it can be pierced by a glass rod), filter with eight layers of gauze, Heat, add 15-20g of agar, continue to heat and mix well, after the agar is dissolved, add 20g of glucose, stir evenly, and then make up water to 1000ml after cooling slightly, the pH is natural. Divide into test tubes or Erlenmeyer flasks, stopper and bandage, sterilize at 121°C for about 20 minutes, take out and shake well, place on an inclined plane or pour on a flat plate, cool and store for later use.

[0042] PDA liquid culture medium: the method is to wash and peel the potatoes first, then weigh 200g of potatoes and cut them into small pieces, add water and boil them (boil for 20-30 minutes, it can be pierc...

Embodiment 3

[0081] Example 3 Research on the NO inhibitory activity of neoandrographolide and its conversion products

[0082] Using RAW264.7 cells to study the NO inhibitory activity of neoandrographolide and its transformation products:

[0083] RAW 264.7 cells were cultured in DMEN high glucose medium (containing penicillin 100U / mL, streptomycin 100μg / mL and 10% fetal bovine serum). The plank density is 1×10 4 unit / ml, CO 2 After incubation in the incubator at 37°C for 4 hours, set up the experimental group (LPS+sample), the control group (untreated), and the LPS group (LPS), add 100 μL of the corresponding solution (LPS concentration is 3 μg / mL), continue to incubate for 36 hours, and measure NO production. Determination method: follow the instructions of the NO detection kit. After adding the drug for 24 hours, the cytotoxic activity was detected by MTT method. NO content by 0,1,2,5,10,20,50, and 100μM NaNO 2 Released NO content of the work curve measured.

[0084] The results...

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Abstract

The invention discloses a creat endophytic fungus strain LM19J01 and application thereof. The strain LM19J01 belongs to fusarium in taxology, is named Fusarium sp., and was collected by China Center for Type Culture Collection on November 11, 2015; and the collection number is CCTCC NO:M2015659. The genome 18s rDNA (ribosomal deoxyribonucleic acid) base sequence of the strain is disclosed as SEQ ID NO.1. The experiment proves that the creat endophytic fungus strain LM19J01 can perform biotransformation on neoandrographolide to obtain the transformed compounds.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to the transformation and application of an endophytic fungus from the plant Andrographis paniculata to diterpenoids of andrographolide. Background technique [0002] Neoandrographolide (Neoandrographolide) is a common andrographolide diterpene lactone. Modern pharmacological studies have shown that neoandrographolide has anti-inflammatory, anti-viral, anti-free radical, anti-HIV and liver-protecting effects. Because it is insoluble in water, it can only be administered orally, which affects the development of preparations. Therefore, it is necessary to carry out structural modification of neoandrographolide to discover andrographolide derivatives with novel structures and to study their biological activities. and application pathways, providing valuable lead compounds for the research of new drugs of Andrographis paniculata. [0003] The current structural modi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12P19/46C12P17/04C12P17/16C07H15/26C12R1/77
CPCC07H15/26C12P17/04C12P17/162C12P19/46C12N1/145C12R2001/77
Inventor 黎万奎王莉莉王杰华张纯唐承晨王吉永
Owner SHANGHAI UNIV OF T C M
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