Artificial recombinant collagen with high expression quantity

Abstract

The invention relates to an artificial recombinant collagen with a high expression quantity. The amino acid sequence of the artificial recombinant collagen is

Description

Technical Field

[0001] The present invention relates to the field of bioengineering, and in particular to artificial recombinant collagen with high expression level. Background Art

[0002] Collagen is a biological macromolecular substance and the main component of animal connective tissue. Collagen is needed in various parts of the human body, such as around the eyes, mouth, armpits, arms, abdomen, etc. It is responsible for maintaining the stability and strength of body tissues by forming a support network on the cell structure. So far, 29 types of collagen have been found in the human body, named I, II, III, IV collagen, etc. in the order of discovery. At present, the market size of collagen products in food functional health products, skin care products, medical devices, especially medical dressings, is gradually increasing.

[0003] Currently, collagen is mainly extracted directly from animal sources. Extracting collagen from animal sources is complicated, and there is a risk of excessive accumulation of toxins, which interferes with the collagen structure and affects the expression of collagen. In addition, the sequence of natural collagen is complex. This complexity may make it difficult to express collagen efficiently during its effectiveness.

[0004] It should be noted that the information disclosed in the above background technology section is only used to enhance the understanding of the background of the present disclosure, and therefore may include information that does not constitute the prior art known to ordinary technicians in the field. Summary of the invention

[0005] In view of the deficiencies of the prior art, the present invention discloses an artificial recombinant collagen with a high expression level.

[0006] The technical solution adopted by the present invention is as follows:

[0007] A high expression level artificial recombinant collagen protein, the amino acid sequence of the high expression level artificial recombinant collagen protein is shown as SEQ ID NO 1.

[0008] Furthermore, the expression yield of the high-expression artificial recombinant collagen is ≥3.0 g / L.

[0009] Furthermore, the expression vector of the high expression amount of artificial recombinant collagen includes Corynebacterium glutamicum.

[0010] The beneficial effects of the present invention are as follows:

[0011] 1. Artificial recombinant collagen has a special sequence, which makes collagen easier to express and greatly improves the yield of recombinant expression and subsequent purification yield.

[0012] 2. The use of artificial recombinant proteins avoids the hidden dangers caused by animal origin or methanol-induced expression, and also reduces the risk of endotoxins in the product. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 This is the secretion expression band diagram of high-expression artificial recombinant collagen.

[0014] Figure 2 This is a bar graph of the fermentation expression yield of high-expression artificial recombinant collagen. DETAILED DESCRIPTION

[0015] The specific implementation of the present invention will be described below in conjunction with the accompanying drawings.

[0016] Embodiment 1:

[0017] A high expression level artificial recombinant collagen, the amino acid sequence of which is shown in SEQ ID NO 1.

[0018] Specific amino acid sequence:

[0019] Galglkgnegppgppgpagspgergpagaagpigipgrpgpqgppgpagekgapgekgpqgpagrdglqgpvglpgpagpvgppgedgdkgeigepgqkgsk gnkgehgppgppgpigpvgqpgaagadgepgargpqghfgakgdegtrgfngppgpiglqglpgpsgekgetgdvgpmgppgppgprgpagpngadgpqgpp gfpgdfgdrgpagldgspglvggtgppgfpglrgsvgpvgpigpagipgpmglsgnkglpgikgdkgeqgtagelgepgypgdkgavglpgppgmrgksgps gppgkdglpghpgqrgevgfqgktgppgppgvvgpqgaagetgpmgerghpgppgppgeqglpgtagkegtkgdpgppgapgkdgpaglrgfpgerglpgta.

[0020] Embodiment 2:

[0021] Expression of high expression levels of artificial recombinant collagen.

[0022] Chassis Cell:

[0023] Platform ID: Bio-52827.

[0024] Specifications: Glycerol / culture.

[0025] Latin genus name: Corynebacterium Glutamicum.

[0026] Strain name: Corynebacterium glutamicum.

[0027] Other numbers: ACCC16522=ATCC13032.

[0028] Medium number: 2.

[0029] Cultivation temperature: 30℃.

[0030] Cultivation time: 24-48 hours.

[0031] Secretory expression results:

[0032] The sequence of the present invention corresponds to Figure 1 Middle band 4, after liquid chromatography detection and calculation, the secretory expression yield of the sequence reached 3.6g / L, and its target band accounted for a larger proportion and less impurity proteins.

[0033] Embodiment 3:

[0034] Fermentation expression experiment of high expression level artificial recombinant collagen.

[0035] Control group: Purchased animal raw collagen from the market.

[0036] Experimental group: artificial recombinant collagen with high expression level was used.

[0037] The experimental group and the control group were subjected to fermentation expression experiments under the same temperature, time and environmental conditions.

[0038] Fermentation expression results:

[0039] like Figure 2 As shown, the expression level of the control group was about 1.0 g / L. The expression level of the experimental group in the first fermentation was about 3.55 g / L; the expression level of the experimental group in the second fermentation was about 3.47 g / L; and the expression level of the experimental group in the third fermentation was about 3.65 g / L.

[0040] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0041] The above-mentioned embodiments only express several implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the invention patent. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. A high expression level artificial recombinant collagen, characterized in that: The amino acid sequence of the high-expression artificial recombinant collagen is shown in SEQ ID NO 1.

2. The high expression level artificial recombinant collagen according to claim 1, characterized in that: The expression yield of the high-expression artificial recombinant collagen is ≥3.0 g / L.

3. The high expression level artificial recombinant collagen according to claim 1, characterized in that: The expression vector of the artificial recombinant collagen with high expression amount comprises Corynebacterium glutamicum.

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