Enzyme composite bactericide with broad-spectrum high-efficient bactericidal action and its preparation method
A technology of preparation and combination of enzymes, applied in the field of food safety and enzyme preparation, replacing antibiotics in feed, to achieve the effect of ensuring food safety, wide bactericidal spectrum and strong bactericidal ability
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Embodiment 1
[0019] The preparation of embodiment 1 enzyme combination bactericidal preparation
[0020] Based on the enzyme activity content in the preparation, lysozyme 400~1000U / g, β-1,3-glucanase 20000~80000U / g, β-mannanase 5000~10000U / g and neutral protease 80000~ 140000U / g combination. Using trehalose 30%-50%, xanthan gum 20%-40%, carboxymethylcellulose sodium 15%-30% and pullulan 5%-10% coating agent, the combined enzyme is prepared into microcapsules Or powdered enzyme combination bactericidal preparation.
[0021] Its preparation method is:
[0022] 1) Preparation of combined enzyme: according to the enzyme activity content of the enzyme, convert the enzyme activity into the required quality, and weigh the corresponding quality of lysozyme, β-1,3-glucanase, β-mannanase and Neutral protease, mix well.
[0023] 2) Enzyme activity protection: using a coating agent formula of 30% to 50% trehalose, 20% to 40% xanthan gum, 15% to 30% sodium carboxymethylcellulose and 5% to 10% pullu...
Embodiment 2
[0026] After Salmonella was cultivated for 5 hours, add 0.5 mL of lysozyme solution of optimal concentration in No. 1 test tube, and add 0.5 mL of enzyme combination bactericide solution in No. 2 test tube (the solid bactericide has been made into a solution, and the enzyme activity of each enzyme is listed in the claims). Within the range described in 1, the same below), add 0.5 mL of phosphate buffer solution to No. 3 test tube, and do three repetitions for each No. tube. Then place it in a water bath for 10 minutes and then take it out. Dilute No. 1 test tube 1000 times, No. 2 test tube 100 times, and No. 3 test tube 100000 times. Take 1 mL of each in a sterile petri dish, pour about 15 mL of nutrient agar medium into it, turn it over after cooling, and place it in a constant temperature incubator at 37 °C. After 48 hours of incubation, the cells were counted. The results are shown in Table 1.
[0027] Table 1 Salmonella plate count results
[0028] project
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Embodiment 3
[0031] After culturing Staphylococcus aureus for 5 hours, add 0.5mL of lysozyme solution with the optimal concentration to No. 1 test tube, 0.5mL enzyme combined fungicide solution to No. 2 test tube, and 0.5mL phosphate buffer solution to No. 3 test tube. repeat. Then place it in a water bath for 10 minutes and then take it out. Dilute No. 1 test tube 1000 times, No. 2 test tube 100 times, and No. 3 test tube 100000 times. Take 1 mL of each in a sterile petri dish, pour about 15 mL of nutrient agar medium into it, turn it over after cooling, and place it in a constant temperature incubator at 37 °C. After 36 hours of incubation, the cells were counted. The results are shown in Table 2.
[0032] Table 2 Plate count results of Staphylococcus aureus
[0033] project
[0034] As can be seen from the results in Table 2, the enzyme combination fungicide has the strongest bactericidal effect, and the number of Staphylococcus aureus is reduced by about 100,000 times com...
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