Bionic anti-caries polypeptide based on casein-rich saliva, derivative, salt and application thereof
A technology of statcasein and peptide derivatives, applied to polypeptide derivatives or their salts and their applications in pharmaceuticals, in the field of biomimetic anti-caries functional polypeptides, can solve problems such as high price, difficulty in extracting natural saliva proteins, and variability , to achieve the effect of low production cost, stable and simple structure, and small molecular weight
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Embodiment 1
[0030] A mineralization-promoting and anti-caries functional polypeptide, the amino acid sequence of which is shown in SEQ ID NO.1;
[0031] The serine phosphorylation modifier of SEQ ID NO.1 is: DpSpSEEKEEEEE.
Embodiment 2
[0032] Example 2 Preparation of Serine Phosphorylated Modifier of SEQ ID NO.1 (hereinafter referred to as DE11)
[0033] 1. Select Fmoc-His(Trt)-Wang Resin as the resin (carrier);
[0034] 2. Fully swell the resin with DCM;
[0035] 3. Use an appropriate concentration of DBLK (hexahydropyridine + DMF) to remove the Fmoc-protecting group;
[0036] 4. Wash several times with DMF to remove DBLK;
[0037] 5. Weigh a suitable condensing agent and activator (HBTU, NMM) and the second Fmoc-protected amino acid (Fomc-Leu-OH) at the C-terminal for coupling;
[0038] 6. The ninhydrin detection method is used for detection to ensure that the connection is relatively complete;
[0039] 7. Wash several times with DMF to wash away the remaining residues and activator condensation agent;
[0040] 8. Carry out coupling according to the amino acid sequence of SEQ ID NO.1, the method refers to steps 3-7;
[0041] 9. After connecting all the amino acids, use steps 3 and 4 to remove the fina...
Embodiment 3
[0045] Example 3 Detection of Polypeptide Nucleation Ability to Hydroxyapatite
[0046] 1. Prepare 50μM polypeptide DE11 solution, add the final concentration of 1.6mM Na 2 HPO 4 and 3.3mM CaCl 2 solution, the pH was adjusted to 7.4, and incubated on a shaker at 37°C for 24h (100 rpm).
[0047] 2. Take 10 μl of the reacted solution and add it dropwise on the copper grid. The negative control is Na without peptide. 2 HPO 4 and CaCl 2 solution. The shape of the precipitate on the copper grid was observed under a transmission electron microscope. figure 1 It showed that the crystals formed in the polypeptide DE11 group were denser than those in the negative control group, showing fascicular or columnar shapes, suggesting that DE11 has a good ability to promote the nucleation and growth of hydroxyapatite.
[0048] 3. Select electron diffraction to show that the crystal precipitate formed by the polypeptide DE11 group has the characteristic diffraction rings 004, 002 and 211...
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