Functional expression of LZ-8, a fungal immunomodulatory protein from Ganoderma lucidium in Pichia pastoris.
LZ-8, a fungal immunomodulatory protein found in Ganoderma lucidium, has been proposed to have therapeutic effects on cancer and autoimmune diseases. To develop an efficient and facile expression system supplying of rLZ-8 of high purity and stable activity for further study and applications, a full-length cDNA of lz8 gene was cloned into the pPIC9K to construct a yeast expression vector pPIC9K-lz8, then transformed to Pichia pastoris strain GS115. The molecular weight of the rLZ-8 was about 17 kDa. The yield of rLZ-8 reached up to 270 mg L(-1) at optimal culture conditions. In vitro, the rLZ-8 was capable of hemagglutinating mouse red blood cells, but no such activity was observed toward human red blood cells, which were similar to the native LZ-8. The mouse splenocyte proliferation activity of rLZ-8 expressed in P. pastoris was much higher than the rLZ-8 expressed in E. coli. All these results suggested that the active rLZ-8 protein can be expressed efficiently in the P. pastoris expression system. This report may supply a new economic and convenient way for the application of LZ-8 protein.
Xue
Qin
Q
College of Life Sciences, Nanjing Agricultural University, Jiangsu, P.R. China.
Ding
Yixin
Y
Shang
Changhua
C
Jiang
Chao
C
Zhao
Mingwen
M
D000818:Animals; D001709:Biotechnology / Q000379:methods; D004912:Erythrocytes; D005260:Female; D005656:Fungal Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D032902:Ganoderma / Q000235:genetics* / Q000378:metabolism; D006384:Hemagglutination; D006801:Humans; D008213:Lymphocyte Activation; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010843:Pichia / Q000235:genetics* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D013154:Spleen / Q000166:cytology / Q000276:immunology
D005656:Fungal Proteins; D011994:Recombinant Proteins; C061148:LZ-8 protein, Ganoderma lucidum