BỘ KHOA HỌC VÀ CÔNG NGHỆ
HỆ THỐNG THÔNG TIN KHOA HỌC VÀ CÔNG NGHỆ

Lọc theo danh mục
  • Năm xuất bản
    Xem thêm
  • Lĩnh vực
liên kết website
Lượt truy cập
 Lượt truy cập :  16,209,983

62.39

Các công nghệ xử lý sinh học, xúc tác sinh học; lên men

Vũ Thị Bích Ngọc; Đỗ Thị Tuyên; Lý Thị Bích Thủy; Quyền Đình Thi; Đỗ Thị Tuyên(1); 

Tối ưu biểu hiện và tinh sạch lumbrokinase trong Pichia pastoris X33

Expression and purification of recombinant Lumbrokinase in Pichia pastoris X33

Công nghệ Sinh học

2014

3

559-564

1811-4989

Lumbrokinase; Biểu hiện; Tinh sạch; Pichia pastoris X33; Nghiên cứu

Lumbrokinase, as a group of potential fibrinolytic enzymes from earthworm, has molecular weights of 25 to 40 kDa. These enzyme was demonstrated capable of directly hydrolyze fibrin, by activating plasminogene into plasmin- the fibrinolytic enzyme of human body. So, lumbrokinase have now been widely using for the treatment of thrombosis. In this study, the gene encoding lumbrokinase from the earthworm Lumbricus rubellus (GeneBank Accession No. AF304199) was codon optimized for expression in P. pastoris X33 using an expression vector pPICZaA. The transformant expressing the highest level of the lumbrokinase (0,767 U/ml) was selected. The recombinant lumbrokinase was produced with the highest level in YPTCM medium after induction of 1.0 percent methanol for 96 h. The recombinant lumbrokinase showed a molecular mass of 47 kDa on SDS-PAGE, specific activity of 2,94 U/mg. The purified of protein was identified with Maldi-Tof mass spectrometry, which showed 100 percent identification to the corresponding peptides of the putative LK from Genbank (code: Q3HRI8). The lumbrokinase was successfully expressed in P. pastoris X33.

TTKHCNQG, CVv 262

Xem toàn văn