Production, formulation, and uses of stable liquid harpin protein formulations
A protease inhibitor, protein technology, applied in the direction of herbicides and algicides, applications, biocides, etc., can solve the problems of limiting the use and versatility of harpin protein
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[0098] Example 1 - Containing Harpin αβ * Preparation of stable liquid compositions of and their efficacy in disease resistance tests
[0099] Recombinant E. coli was used to express harpin αβ (superharpin of SEQ ID NO: 1 ) under the control of a constitutive promoter. After fermentation, the suspension was diluted in 2 volumes of potassium phosphate buffer to adjust the pH of the suspension to 7.0.
[0100] The resulting solution was heat-treated at 95°C for 1 minute using a heat exchange system and then cooled to 45°C within 40 minutes.
[0101] Lysozyme was added to the solution after reaching about 38-42°C, mixed to a final concentration of 1 ppm, and allowed to react for 45 minutes. This facilitates the rupture of the cell wall of the bacteria, resulting in the formation of a crude harpin αβ extract.
[0102] The resulting crude extract was then centrifuged for 5 minutes to remove some cellular debris. The resulting supernatant was separated and subjected to further h...
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