Recombination human collagen and biological synthesis method thereof

A human-like collagen, biosynthetic technology, applied in the biological field, can solve the problem of not being able to form Hyp residues, achieve high thermal denaturation temperature and thermal decomposition temperature, high thermal stability, and stable structure

Inactive Publication Date: 2008-03-26
ZHEJIANG SCI-TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in most E. coli and yeast, they cannot syn

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  • Recombination human collagen and biological synthesis method thereof
  • Recombination human collagen and biological synthesis method thereof
  • Recombination human collagen and biological synthesis method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1: Recombinant human-like collagen containing a non-collagen region at the carboxy-terminal

[0050] The recombinant human-like collagen in this example is composed of a collagen region and a non-collagen region; the non-collagen region is located at the carboxyl terminal of the collagen region, and the collagen region is expressed after 8 repeated polymerizations of the target DNA monomer. The structural features of the designed sequence are: a β-helix-like trimer formed in the non-collagen region, and this trimer with a β-hairpin shape forms a β-sheet, and this region has a strong hydrophobic effect. In the collagen molecule During the assembly process, it can play the role of a node, twisting the peptide chains of the three collagen regions together to form a triple superhelical structure, thereby improving the thermal stability of the recombinant human-like collagen.

[0051] A designed recombinant human-like collagen has the amino acid sequence shown in SEQ...

Embodiment 2

[0105] Example 2: Experimental research on recombinant human-like collagen

[0106] (1) Put a large amount of expressed and purified recombinant humanoid protein into a dialysis bag, put it into distilled water for dialysis for three days, and change the distilled water intermittently at an appropriate time to remove a large amount of salt ions.

[0107] (2) Properly concentrate the protein solution after dialysis, put it into a -80°C refrigerator for pre-cooling for more than 2 hours, put it into a freeze dryer, and freeze-dry it for 20-24 hours to obtain recombinant human-like collagen.

[0108] (3) Take 1-5 mg of recombinant human-like collagen for differential scanning calorimetry (DSC) (Figure 3), thermogravimetric (TGA) and micro-thermogravimetric (DTG) analysis (Figure 4-7).

[0109] Fig. 3 is the differential scanning calorimetry (DSC) spectrogram of the recombinant human-like collagen with different molecular weights of the embodiment of the present invention and the ...

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Abstract

The present invention discloses one kind of recombinant analogous human collagen and its biological synthesis process. The recombinant analogous human collagen consists of one collagen region and one non-collagen region. The non-collagen region in the carboxyl group end of the collagen region has one amino acid sequence originated the minor fibrin of T4 bacteriophage and as shown in SEQ ID No. 3, and the collagen region has the amino acid sequence shown in SEQ ID No. 2. The recombinant analogous human collagen has relatively high heat stability, high hydrophilicity, high biocompatibility, heat denaturation temperature and heat decomposition temperature higher than that of natural collagen, and stable structure. It may be applied widely in biomedicine material, tissue engineering, cosmetics, food and other fields.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a recombinant human-like collagen with high thermal stability and a biosynthetic method thereof. Background technique [0002] Collagen is very rich in mammalian body, it is a kind of fibrous protein in extracellular matrix, it is very important for the formation of animal and human skin, blood vessels, tendons and cartilage, and it provides certain support for these connective tissues Structural and mechanical properties. On the other hand, collagen has good cell adhesion properties, and the combination with cells has a certain positive effect on cell growth, differentiation and migration. In addition, it also has a certain effect on platelet aggregation. In view of the inherent biocompatibility, biodegradability and absorbability of collagen, as well as the promotion of cell formation and many other functions, collagen has a wide range of application values ​​in the fields of bio...

Claims

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Application Information

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IPC IPC(8): C07K19/00C07K14/78C12N15/62
Inventor 姚菊明杜春玲
Owner ZHEJIANG SCI-TECH UNIV
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