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Recombinant human III type collagen and application thereof

A collagen and recombinant carrier technology, applied in the field of recombinant human type III collagen, can solve the problem of difficulty in obtaining Hyp

Pending Publication Date: 2022-04-05
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although Hyp plays a very important role in the stabilization of the collagen triple helix structure, in many cases, Hyp is still very difficult to obtain

Method used

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  • Recombinant human III type collagen and application thereof
  • Recombinant human III type collagen and application thereof
  • Recombinant human III type collagen and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] experimental method:

[0035] (1) Construction of recombinant strains:

[0036] Utilize the target gene shown in the chemically synthesized SEQ ID NO.2, introduce EcoR I and Not I double restriction site, insert in the expression vector pPIC9K, obtain recombinant plasmid (structural schematic diagram sees figure 1 ), linearized the recombinant plasmids and electrotransformed them into Pichia pastoris GS115 respectively, after colony PCR identification, Mut phenotype screening, and G418 copy number screening, the expression was induced with methanol.

[0037] (2) Obtained from recombinant human type III collagen:

[0038] BMGY medium was used to culture overnight at 28°C and 250rpm, and then transferred to BMMY medium the next day, cultured at 28°C and 250rpm for 96h, and methanol was added every 24h. The bacteria and fermentation supernatant were collected by centrifugation.

[0039] (3) Disintegration of recombinant yeast cells:

[0040]The fermentation broth was c...

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PUM

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Abstract

The invention relates to the technical field of biology, in particular to recombinant human III type collagen and application thereof. The human full-length III-type collagen is optimized, a segment containing 123 amino acid sequences is selected, a tripeptide sequence Gly-Pro-Hyp in the segment sequence is directionally replaced with a Gly-Pro-Arg sequence, the Gly-Pro-Arg sequence is subjected to four repetitions, a GPPGPCCGGG sequence is connected to the-C tail end, and the recombinant protein has a three-level helical structure, shows higher stability, and can be used for preparing the recombinant collagen. And the collagen has properties close to those of natural collagen. The recombinant human III-type collagen can be effectively, stably and massively expressed in pichia pastoris, and meanwhile, a C terminal is connected with six histidine specific affinity purification tags, so that a high-purity product is easy to obtain, and the recombinant human III-type collagen can be suitable for industrial large-scale production.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a recombinant human type III collagen and its application. Background technique [0002] Collagen, also known as collagen, is a glycoprotein. Collagen in human skin is type I collagen and type III collagen. Type I collagen mainly exists in adult skin, tendon, and bone tissue, and type III collagen mainly exists in infant skin or blood vessels. Intima, intestinal tract, type I collagen and type III collagen are closely related to the skin damage repair process and repair quality. Type III collagen has excellent biological activity and tissue compatibility, can promote the proliferation of fibroblasts in the dermis, increase cell activity, and be absorbed by fibroblasts as raw materials for synthesizing collagen, stimulating cells to synthesize more collagen , Fill and repair the damaged and aging skin, rebuild the network structure, enhance the expansion force of the damaged skin, an...

Claims

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Application Information

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IPC IPC(8): C07K14/78C12N15/12C12N15/81C12N1/19A61K8/65A61Q19/00A61Q19/08C12R1/84
Inventor 陈海佳陈东煌姜交华
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD