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Extracellular polysaccharide-containing extract, exopolysaccharide and composition

A technology of exopolysaccharides and extracts, applied in the field of extracts containing exopolysaccharides, can solve the problems of low yield and limited market application, and achieve excellent synergistic effects

Pending Publication Date: 2022-03-11
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the low production of bacterial exopolysaccharides, its market application is limited

Method used

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  • Extracellular polysaccharide-containing extract, exopolysaccharide and composition
  • Extracellular polysaccharide-containing extract, exopolysaccharide and composition
  • Extracellular polysaccharide-containing extract, exopolysaccharide and composition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0097] 1.1 Extraction of crude exopolysaccharide (i.e. exopolysaccharide extract)

[0098] (1) Inoculate 10 μL of glycerol bacteria of Bacillus thuringiensis 4D19 into 5 mL of liquid LB medium, and activate at 30°C and 220 r / min for 10 hours to obtain activated bacterial liquid;

[0099] (2) Transfer the activated bacterial solution to 100mL liquid LB culture medium and the liquid sugar-producing medium whose substrate is glucose with a 1% inoculation amount, and cultivate it in a 500mL Erlenmeyer flask for 30 hours;

[0100] (3) Collect the bacterial solutions cultured in the three mediums respectively, adjust the pH of the bacterial solutions to 8.5 with 1mol / L NaOH, centrifuge at 4°C, 7000r / min for 30min, and collect the first supernatant;

[0101] (4) Add 1 / 10 volume of 5wt% trichloroacetic acid aqueous solution to the first supernatant, and denature the protein for 2 hours at 25°C;

[0102] (5) 4°C, 7000r / min, centrifuge for 30min, collect the second supernatant, adjust ...

Embodiment 2

[0112] Crude exopolysaccharide extraction is the same as the first section 1.1 in Example 1, the difference is:

[0113] In step (2), the activated bacterial liquid was transferred to 100 mL liquid LB medium and sucrose-based liquid sugar-producing medium with an inoculation amount of 1%, and cultivated in a 500 mL Erlenmeyer flask for 30 h;

[0114] In step (3), collect the bacterial liquid cultured in the three mediums respectively, adjust the pH of the bacterial liquid to 8 with 1mol / L NaOH, centrifuge at 3°C, 6000r / min for 40min, and collect the first supernatant ;

[0115] In step (4), add 1 / 10 volume of 4wt% trichloroacetic acid aqueous solution to the first supernatant, and denature the protein at 35°C for 3 hours;

[0116] In step (5), centrifuge at 3°C, 8000r / min for 30min, collect the second supernatant, and adjust the pH to 6;

[0117] In step (6), add 3 times the volume of absolute ethanol and precipitate in a refrigerator at 2°C for 16 hours;

[0118] In step (...

Embodiment 3

[0121] Crude exopolysaccharide extraction is the same as the first section 1.1 in Example 1, the difference is:

[0122] In step (3), collect the bacterial liquids cultured in the three mediums respectively, adjust the pH of the bacterial liquids to 9 with 1mol / L NaOH, centrifuge at 8000r / min at 10°C for 30min, and collect the first supernatant ;

[0123] In step (4), add 1 / 10 volume of 5wt% trichloroacetic acid aqueous solution to the first supernatant, and denature the protein at 25°C for 1.5 hours;

[0124] In step (5), centrifuge at 3°C, 6000r / min for 40min, collect the second supernatant, and adjust the pH to 7;

[0125] In step (6), add 3 times the volume of absolute ethanol to precipitate in a refrigerator at 8°C for 20 hours;

[0126] In step (7), centrifuge at 10° C. and 8000 r / min for 30 minutes to discard the supernatant, and freeze-dry the precipitate to obtain crude exopolysaccharide powder.

[0127] After quantification, the amount of exopolysaccharide in the ...

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Abstract

The invention relates to an extracellular polysaccharide-containing extract, extracellular polysaccharide and a composition, the extract is extracted from bacillus thuringiensis, the main component of the extract is extracellular polysaccharide, and the extracellular polysaccharide has a synergistic effect on bacillus thuringiensis insecticidal crystal protein and / or bacillus thuringiensis vegetative insecticidal protein.

Description

technical field [0001] The invention relates to an extract containing extracellular polysaccharide, which is extracted from Bacillus thuringiensis, mainly composed of extracellular polysaccharide, and has a synergistic effect on the insecticidal crystal protein of Bacillus thuringiensis. Background technique [0002] Bacterial exopolysaccharide is a polysaccharide mixture synthesized and secreted by bacteria during growth and metabolism. It is a long-chain polymer formed by the polymerization of the same or different monosaccharide units through glycosidic bonds. Extracellular polysaccharides (EPSs) produced by bacteria mainly exist in two forms, capsular polysaccharides (Capsular polysaccharides, CPS) tightly bound to the cell surface and mucus polysaccharides ("Slime polysaccharides" secreted outside the cells in the form of mucus). ”), also known as free exopolysaccharide (Free EPS), in which a small amount of free exopolysaccharide is the main component of biofilm as a w...

Claims

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Application Information

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IPC IPC(8): C08B37/00C12P19/02C12P19/00A01N43/16A01N63/50A01N63/23A01P7/04C12R1/07
CPCC08B37/006C08B37/00C08B37/0003C12P19/04A01N43/16A01N63/50A01N63/23
Inventor 耿丽丽王美玲张杰束长龙王泽宇
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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