A kind of preparation method of ferrous chelating antimicrobial peptide
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An antibacterial peptide and chelation technology, applied in the field of preparation of ferrous chelated antibacterial peptides, can solve the problems of high price and large side effects, and achieve the effects of convenient operation, large-scale deep processing, and abundant scraps.
Inactive Publication Date: 2015-11-25
ZHEJIANG OCEAN UNIV
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There are two types of iron supplements currently in use, chemically synthesized iron supplements and biological preparations. The former has a lot of side effects, while the latter is expensive
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Embodiment 1
[0021] Embodiment 1 Preparation of hairtail enzymolysis protein ferrous chelating antimicrobial peptide
[0022] 1. The hairtail leftovers are crushed by a minced meat grinder, take 10g and add distilled water 100mL, then add 200mg into papain, carry out enzymatic hydrolysis at 40°C for 8-10h, heat to 90-100°C, inactivate the enzyme for 10-20min, 3000g After centrifuging for 10 min, filter the surface oil with filter paper, and then filter the precipitate.
[0023] 2. Add ferrous chloride to the enzymolysis solution, the ratio of ferrous chloride to the enzymolysis solution by volume is 1-30:40 (mg:mL), shake and chelate for 30-50°C at 30-50°C After 60 minutes, centrifuge and remove the precipitate to become the crude extract of hairtail enzymolyzed protein ferrous chelated antimicrobial peptide.
[0024] 3. Vacuum concentrate the crude extract of hairtail enzymatic protein ferrous chelated antibacterial peptide, concentrate at 40°C for 8 hours; freeze-dry the concentrate at ...
Embodiment 2
[0029] Antibacterial experiment of embodiment 2 hairtail enzymolysis protein ferrous chelating antimicrobial peptide
[0030] Using the Oxford cup method, the antibacterial activity experiment was carried out on the hairtail enzymatic protein ferrous chelating antimicrobial peptide of Example 1. The bacteria was Staphylococcus aureus ATCC25923.
[0031] The minimum inhibitory concentration of hairtail enzymatic protein ferrous chelated antimicrobial peptide against Staphylococcus aureus is shown in Table 1, and the results of minimum bactericidal concentration are shown in Table 2.
[0032] Table 1 Minimum inhibitory concentration of ferrous chelated antimicrobial peptides from hairtail enzymatic protein
[0033]
[0034] Note: "+" means colony growth, "-" means no colony growth;
[0035] Table 2 Minimum bactericidal concentration of ferrous chelated antimicrobial peptides from hairtail enzymatic protein
[0036]
[0037] Note: "+" means there is colony growth, "-" me...
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Abstract
The invention discloses a preparation method for a ferrous chelated antibacterial peptide, especially to a preparation method for a hairtail enzymatic hydrolysis protein ferrous chelated antibacterial peptide. The method comprises the following steps: 1) adding leftovers of hairtail into distilled water, then adding papain for enzymatic hydrolysis and adjusting the pH value of enzymatic hydrolysis liquid to be 5.5 to 6.5; and 2) adding ferrous chloride into the enzymatic hydrolysis liquid, carrying out oscillating chelation for 30 to 60 min at a chelation temperature of 30 to 50 DEG C, then carrying out centrifugation and removing a precipitate so as to obtain a crude extract of the hairtail enzymatic hydrolysis protein ferrous chelated antibacterial peptide. To obtain a refined product of the hairtail enzymatic hydrolysis protein ferrous chelated antibacterial peptide, the crude extract undergoes the following purification steps: 1) freeze drying the crude extract of the hairtail enzymatic hydrolysis protein ferrous chelated antibacterial peptide, eluting and separating the crude extract by using sephadex chromatography, subjecting chromatographic components with antimicrobial activity to reverse high performance liquid chromatographic elution and collecting eluate; and 2) subjecting the eluate to vacuum freeze drying so as to obtain the hairtail enzymatic hydrolysis protein ferrous chelated antibacterial peptide. The method provided by the invention has the characteristics of simple operation, a high separation and recovery rate and low cost and enables high value utilization of waste produced in processing of hairtail to be realized.
Description
technical field [0001] The invention relates to a preparation method of a ferrous chelating antibacterial peptide, in particular to a preparation method of a hairtail proteolytic hydrolysis of a ferrous chelating antibacterial peptide. Background technique [0002] Hairtail is one of the most important marine economic fish in our country. Hairtail is rich in nutrition and delicious. In addition to being eaten directly, hairtail can also be processed into canned food and instant food. However, the head, bones, and viscera of hairtail will be discarded during processing. If not used, this part accounts for more than half of the weight of hairtail. At present, the utilization of these leftovers is mostly focused on the production of animal feed, fish sauce, hydrolyzed protein, etc., but these leftovers are rich in unsaturated fatty acids and high-quality protein resources. Such processing methods not only waste high-quality protein resources, but also increase production costs....
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