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Method for extracting natural green pigment in metarhizium anisopliae

An extraction method, the technology of Metarhizium anisopliae, applied in the direction of using spores, methods based on microorganisms, natural dyes, etc., can solve problems such as unstable properties

Active Publication Date: 2021-05-11
ANHUI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Moreover, natural chlorophyll is easily destroyed by factors such as light, temperature, and oxidants in the environment after detaching from plant cells, and its properties are unstable. Therefore, there is an urgent need for a natural green pigment with better environmental tolerance as a substitute for chlorophyll

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] The concrete extraction operation steps of natural green pigment in Metarhizium anisopliae are as follows:

[0040] 1.1 Culture strains

[0041] Metarhizium anisopliae ( M. anisopliae ) were inoculated in PDA medium, and cultured at 20°C for 3 days to obtain solid strains.

[0042] The formula of PDA medium is: 6g potato dry powder, 20g glucose, 15g agar, add water to 1000mL.

[0043] 1.2 Expansion of cultivation

[0044] Put the solid bacteria obtained in step (1) into the PDB liquid medium, culture in the Erlenmeyer flask at 150 rpm, 21°C for 2 days, and inoculate rice at a volume-to-weight ratio (V / W) of 1%. culture medium for 8 days to obtain a rice culture.

[0045] The formula of PDB medium: 6g potato powder, 20g glucose, add water to 1000mL; rice medium is rice soaked in water for 10mim and autoclaved.

[0046] 1.3 Spore collection

[0047] The rice culture was dried at -50°C, and then the spores were harvested using a spore collector.

[0048] The spore c...

Embodiment 2

[0064] The specific extraction operation steps of the green pigment in Metarhizium anisopliae are as follows:

[0065] 2.1 Culture strains

[0066] Metarhizium anisopliae Roberts ( M. Robberstii )) were inoculated in PDA medium and cultured for 7 days at 30°C to obtain solid strains.

[0067] The formula of PDA medium is: 20g peeled fresh potato, 20g glucose, 15g agar, add water to 1000mL.

[0068] 2.2 Expansion of cultivation

[0069] Put the solid bacteria obtained in step (1) into the PDB liquid medium, culture in the Erlenmeyer flask at 250 rpm and 30°C for 6 days, and inoculate rice at a volume-to-weight ratio (V / W) of 10% culture medium for 16 days to obtain a rice culture.

[0070] The formula of PDB medium: 20g peeled fresh potatoes, 20g glucose, add water to 1000mL; rice medium is rice soaked in water for 10mim and autoclaved.

[0071] 2.3 Spore collection

[0072] The rice culture was dried at 130°C, and then the spores were collected using a cyclone separator...

Embodiment 3

[0088] The concrete extraction operation steps of natural green pigment in Metarhizium anisopliae are as follows:

[0089] 3.1 Culture strains

[0090] Metarhizium anisopliae ( M. acridum ) were inoculated in PDA medium and cultured at 25°C for 5 days to obtain solid strains.

[0091] The formula of PDA medium is: 6g potato dry powder, 20g glucose, 15g agar, add water to 1000mL.

[0092] 3.2 Expansion of cultivation

[0093] Put the solid bacteria obtained in step (1) into the PDB liquid medium, culture it in the Erlenmeyer flask at 200 rpm and 26°C for 4 days, and inoculate it at a volume-to-weight ratio (V / W) of 5%. The rice culture medium was cultured for 12 days to obtain a rice culture.

[0094] The formula of PDB medium: 6g potato powder, 20g glucose, add water to 1000mL; rice medium is rice soaked in water for 10mim and autoclaved.

[0095] 3.3 Spore collection

[0096] The rice culture was dried at 50°C, and then the spores were collected using a cyclone separat...

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Abstract

The invention relates to a method for extracting natural green pigment in metarhizium anisopliae, and belongs to the technical field of extraction and preparation of biological products. The method comprises the following operation steps: culturing a strain by using a fungus of Metarhizium, carrying out enlarged culture and collecting spores, removing impurities from the spores, preparing a pigment crude extract, concentrating and drying, refining pigment precipitates, and drying the refined pigment precipitates to obtain the dark green powdery natural green pigment. The method solves the problem that green fungal pigments of Metarhizium anisopliae cannot be extracted, and lays a foundation for further structural identification and utilization. The stability of the natural green pigment extracted by the method is superior to that of chlorophyll commonly used in the natural green pigment at present, and the method has a wide application prospect. The process is simple, convenient and stable, the investment scale of production equipment is controllable, production is flexible, and large-scale production can be achieved.

Description

technical field [0001] The invention belongs to the technical field of extraction and preparation of biological products, and in particular relates to a process for extracting green pigment from metarhizium anisopliae spores. Background technique [0002] The spores of Metarhizium anisopliae appear green, but the green pigment has not been isolated and identified so far. The main reason is that the commonly used natural pigment extraction methods cannot extract the green pigment from Metarhizium anisopliae. Only by extracting the green pigment can the identification of its molecular structure and metabolic pathway, and the foundation for its commercial application be laid. [0003] At present, the harmful effects of synthetic pigments on consumers and the environment are widely concerned, and seeking natural pigments as substitutes for synthetic pigments is becoming a research hotspot. Worldwide demand for natural colorants is growing rapidly in the food, cosmetic and texti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C09B67/54C09B61/00C12N1/14C12N3/00C12R1/645
CPCC09B67/0096C09B61/00C12N1/14C12N3/00
Inventor 胡丰林陆瑞利尉杰赵铖
Owner ANHUI AGRICULTURAL UNIVERSITY
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