Title:
GENETICALLY ENGINEERED BACTERIUM FOR PRODUCING PORCINE MYOGLOBIN, AND FERMENTATION AND PURIFICATION OF SAME
Document Type and Number:
WIPO Patent Application WO/2022/242033
Kind Code:
A1
Abstract:
Provided in the present invention are a genetically engineered bacterium for producing porcine myoglobin, and methods for fermenting and purifying same. According to the present invention, the gene of the porcine myoglobin is integrated into Pichia pastoris, thereby realizing efficient secretion and expression of the porcine myoglobin. On this basis, a medium and culture conditions of recombinant Pichia pastoris are optimized, which can significantly increase the yield of the porcine myoglobin, so that the yield can reach 285.42 mg/L under fermentation tank conditions. Creatively, different concentrations of ammonium sulfate are added into a fermentation broth step by step, so that the final myoglobin obtained by means of concentration has a purity of 88.04% and a purification rate of 66.05%. According to the present invention, the highly efficient expression and high-level purification of the porcine myoglobin are realized by means of the synthesis, fermentation, purification and other steps performed on the porcine myoglobin, which provides wide prospects for industrial production of the porcine myoglobin.
Inventors:
ZHAO XINRUI (CN)
ZHANG BOHAN (CN)
ZHOU JINGWEN (CN)
DU GUOCHENG (CN)
LI JIANGHUA (CN)
CHEN JIAN (CN)
YU FEI (CN)
LU WEI (CN)
QIAN YUAN (CN)
ZHANG BOHAN (CN)
ZHOU JINGWEN (CN)
DU GUOCHENG (CN)
LI JIANGHUA (CN)
CHEN JIAN (CN)
YU FEI (CN)
LU WEI (CN)
QIAN YUAN (CN)
Application Number:
PCT/CN2021/126637
Publication Date:
November 24, 2022
Filing Date:
October 27, 2021
Export Citation:
Assignee:
JIANGSU DONGHUI BIO TECH CO LTD (CN)
UNIV JIANGNAN (CN)
UNIV JIANGNAN (CN)
International Classes:
C12N15/81; C07K1/16; C07K1/18; C07K1/30; C07K1/36; C07K14/805; C12N1/16; C12N1/19; C12N1/38; C12N15/12; C12P21/02; C12R1/84
Foreign References:
| CN113186147A | 2021-07-30 | |||
| CN113150120A | 2021-07-23 | |||
| CN113265346A | 2021-08-17 | |||
| CN105745332A | 2016-07-06 |
Other References:
G DODSON , R E HUBBARD, T J OLDFIELD, S J SMERDON, A J WILKINSON: "Apomyoglobin as a molecular recognition surface: expression, reconstitution and crystallization of recombinant porcine myoglobin in Escherichia coli", PROTEIN ENGINEERING, OXFORD UNIVERSITY PRESS, SURREY., GB, vol. 2, no. 3, 30 September 1988 (1988-09-30), GB , pages 233 - 237, XP009541277, ISSN: 0269-2139, DOI: 10.1093/protein/2.3.233
DATABASE Protein ANONYMOUS : "myoglobin [Sus scrofa] ", XP093007725, retrieved from NCBI
DATABASE Protein ANONYMOUS : "myoglobin [Sus scrofa] ", XP093007725, retrieved from NCBI
Attorney, Agent or Firm:
HARBIN SHINEIP INTELLECTUAL PROPERTY AGENCY CO., LTD. (CN)
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