Title:
PICHIA PASTORIS ENGINEERING STRAIN FOR RECOMBINANT TYPE I COLLAGEN, CONSTRUCTION METHOD THEREFOR AND USE THEREOF
Document Type and Number:
WIPO Patent Application WO/2023/221787
Kind Code:
A1
Abstract:
Disclosed are a pichia pastoris engineering strain for a recombinant type I collagen, a construction method therefor and use thereof. A gene sequence for a type I humanized collagen α1 chain mature peptide is synthesized and inserted downstream of a Kex2 protease recognition site in an α-factor secretion signal peptide of a vector to construct a recombinant plasmid pPICZalphaA-col(I)α1. The plasmid is introduced into pichia pastoris X-3, and screening is performed to obtain a recombinant strain. The present invention further provides a method for preparing a type I humanized collagen using the recombinant strain. An improved fermentation culture medium and a high-density fermentation method are provided, which allow for an expression level of up to 6 g/L or above. The purified recombinant I-type collagen has a purity of up to 98% or above, and is 100% identical to the corresponding part of an amino acid sequence of a human type I collagen. The production process is green and environment-friendly. The product has relatively high biocompatibility and bio-safety. The product can be applied to a wide range of fields such as cosmetic medicine, medical instruments, and medical biological materials.
Inventors:
ZHENG HAIZHOU (CN)
JIANG YAN (CN)
WEI JINGSHUANG (CN)
SHEN XUDONG (CN)
PAN YONGGANG (CN)
HE LI (CN)
CHANG LIANG (CN)
WANG XIN (CN)
LIU ZHEN (CN)
HE QINGSHENG (CN)
CHEN LIHUA (CN)
GAO JIAN (CN)
JIANG YAN (CN)
WEI JINGSHUANG (CN)
SHEN XUDONG (CN)
PAN YONGGANG (CN)
HE LI (CN)
CHANG LIANG (CN)
WANG XIN (CN)
LIU ZHEN (CN)
HE QINGSHENG (CN)
CHEN LIHUA (CN)
GAO JIAN (CN)
Application Number:
PCT/CN2023/092412
Publication Date:
November 23, 2023
Filing Date:
May 06, 2023
Export Citation:
Assignee:
NORTH CHINA PHARMACEUTICAL GROUP NEW DRUG RES AND DEVELOPMENT CO LTD (CN)
International Classes:
C12N1/19; C07K1/18; C07K1/30; C07K1/36; C07K14/78; C12N15/12; C12N15/81; C12P21/02
Domestic Patent References:
| WO2021258650A1 | 2021-12-30 | |||
| WO2014170460A2 | 2014-10-23 |
Foreign References:
| CN112626074A | 2021-04-09 | |||
| CN102747097A | 2012-10-24 |
Attorney, Agent or Firm:
SHIJIAZHUANG GUOYU PATENT & TRADEMARK AGENCY (CN)
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