Title:
LIVE CELL LIPOSOME OF RHODOTORULA GLUTINIS CARRYING EXOGENOUS POLYPEPTIDE OF RECOMBINANT PHOSPHOCHOLINE CYTIDINE TRANSFERASE AND USE THEREOF
Document Type and Number:
WIPO Patent Application WO/2020/082221
Kind Code:
A1
Abstract:
Disclosed are a live cell liposome of recombinant Rhodotorula glutinis carrying an exogenous polypeptide and a use thereof. An integrative expression vector for an exogenous phosphocholine cytidine transferase gene CCT is constructed by homologous recombination. With a CCT gene recombinant engineered strain as a carrier, an exogenous polypeptide is successfully introduced into the recombinant engineered strain. The beneficial effects are as follows: Stable and prolonged expression of CCT is achieved, and the expression level of CCT increases by about 1.5 times. Compared with a wild-type strain, the contents of lecithin and fatty acids increase significantly. Exogenous polypeptide is introduced into cells of the strain by electroporation. The exogenous polypeptide enters lipid droplets in the recombinant strain by virtue of the lipophilicity thereof, thus achieving successful encapsulation of the exogenous polypeptide. Oral administration of the whole insulin-carrying GM4-CCT-insulin strain to mice for 7 days revealed an increase in insulin content in the serum and important organs (liver, lung, spleen, and kidney) of the mice. Oral administration of the GM4-CCT-insulin strain to diabetic mice revealed significant decrease in blood sugar, suggesting a marked blood sugar-lowering effect of GM4-CCT-insulin as an exogenous insulin.
Inventors:
LI SHIYU (CN)
SUN HANXIAO (CN)
WANG JIAJIA (CN)
SUN HANXIAO (CN)
WANG JIAJIA (CN)
Application Number:
PCT/CN2018/111320
Publication Date:
April 30, 2020
Filing Date:
October 23, 2018
Export Citation:
Assignee:
LI SHIYU (CN)
International Classes:
C12N1/19; C12N15/54; C12N15/81
Foreign References:
| CN107345211A | 2017-11-14 | |||
| CN107400635A | 2017-11-28 | |||
| CN103249834A | 2013-08-14 | |||
| CN102803289A | 2012-11-28 | |||
| CN107236679A | 2017-10-10 |
Other References:
FRIESEN, J. A. ET AL.: "Purification and Kinetic Characterization of CTP: Phosphocholine Cytidylytransferase from Saccharomyces Cerevisiae", PROTEIN EXPRESSION AND PURIFICATION., vol. 21, no. 1, February 2001 (2001-02-01), pages 141 - 148, XP055707479, DOI: 20190701174832A
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