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Recombinant humanized collagen and its preparation method

A technology of human collagen and original domain, which is applied in the field of highly hydrophilic recombinant human collagen and its preparation, can solve the problems of low biological activity of the expressed product, imperfect processing and modification system, difficult application of the expressed product, etc., to achieve Conducive to separation and purification, lower production costs, and easier feasibility

Active Publication Date: 2013-10-30
JIANGSU JLAND BIOTECH CO LTD
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Problems solved by technology

Bacterial expression systems generally produce pyrogens, which make the expression products difficult to apply clinically. Proteins are often expressed in the form of inclusion bodies, which makes product purification difficult, and the post-translational processing and modification system of the prokaryotic expression system is imperfect. The biological activity of the expressed products low-level deficiencies

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  • Recombinant humanized collagen and its preparation method
  • Recombinant humanized collagen and its preparation method
  • Recombinant humanized collagen and its preparation method

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Embodiment Construction

[0021] The recombinant human collagen of the present invention has an amino acid sequence such as SEQ NO.3, which contains 599 amino acids and a molecular weight of 55.0 kDa.

[0022] The preparation method of the above-mentioned recombinant human collagen, the steps are as follows:

[0023] (1) Construction of genetically engineered bacteria expressing recombinant human collagen to obtain Pichia pastoris genetically engineered bacteria, the construction of which Pichia pastoris genetically engineered bacteria is as follows:

[0024] 1) Based on the tripeptide repeat sequence characteristics of human type III collagen α1 chain collagen domain Gly-X-Y (glycine-X-Y), design and artificially synthesize a segment of human collagen gene monomer Gel, its nucleotide sequence is as shown in SEQNO.1 Then, through in vitro enzyme digestion and connection, using Escherichia coli as a cloning host, constructing an expression vector containing a six-tandem human collagen gene monomer in th...

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Abstract

The invention discloses a recombinant humanized collagen. Its amino acid sequence is as shown in SEQNO.3 and has 599 amino acids. The molecular weight is 55.0kDa. A preparation method of the recombinant humanized collagen comprises the following steps of: constructing a gene engineering strain for the expression of the recombinant humanized collagen to obtain a Pichia pastoris engineering strain; carrying out fermentation culture on the engineering strain, and realizing inducible expression of the recombinant humanized collagen to obtain a recombinant humanized collagen fermentation broth; and purifying the fermentation broth to obtain the recombinant humanized collagen. The humanized collagen gene Ge1 designed in the invention is a brand-new sequence which is greatly shorter than a natural humanized collagen gene (Kbp) in length. By the preparation method, operation simplification at the molecular level is realized, and the gene engineering strain fermentation production of the collagen is more easily realized. The humanized collagen gene Ge1 contains characteristics of a natural humanized collagen gene. And simultaneously, its expressed protein has excellent characteristics of a natural humanized collagen.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a highly hydrophilic recombinant human collagen and a preparation method thereof. Background technique [0002] Collagen (also known as collagen) is the most abundant protein family in animals. Collagen has strong biological activity and biological function, can participate in cell migration, differentiation and reproduction, and makes connective tissue have mechanical strength. At the same time, collagen can also promote cell growth, and has hemostatic, biocompatibility and biodegradation properties. Based on these characteristics, collagen has a wide range of applications in medical and health fields such as burns, trauma, corneal diseases, health care, beauty, orthopedics, hard tissue repair, wound hemostasis, drug delivery, and sustained release technology. In addition, collagen can also be used as food additives, feed additives, flocculants, photographic emulsion materials, sur...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/78C12N15/12C12N15/81C12P21/02C12R1/84
Inventor 杨树林刘斌高立虎李冰雷云霆张静胡利唐启伟
Owner JIANGSU JLAND BIOTECH CO LTD
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