Efficient production of recombinant human pleiotrophin in yeast, Pichia pastoris.
Approximately 260 mg/l of authentic recombinant human pleiotrophin (rhPTN) was expressed into the medium of high-cell density fermentation using a Pichia pastoris protein expression system. The prepro-sequence of yeast alpha-mating factor was used successfully. The recombinant hPTN was efficiently recovered from the medium by expanded bed adsorption, and purified using successive column chromatography steps. In the purified rhPTN preparation, modified rhPTN were scarcely detected. Circular dichroism measurement of the purified PTN showed the presence of the characteristic beta-structures in the protein.
Murasugi
Akira
A
Meiji Institute of Health Science, Meiji Milk Products Co., Ltd., Naruda, Odawara, Kanagawa, Japan. AKIRA_MURASUGI@MEIJI-MILK.COM
Kido
Isao
I
Kumai
Hideshi
H
Asami
Yukio
Y
D002352:Carrier Proteins / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods*; D016207:Cytokines / Q000235:genetics*; D005285:Fermentation; D006801:Humans; D000072235:Mating Factor; D010455:Peptides / Q000235:genetics; D010843:Pichia / Q000235:genetics*; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D019076:Transgenes
D002352:Carrier Proteins; D016207:Cytokines; D010455:Peptides; D011994:Recombinant Proteins; C066845:pleiotrophin; D000072235:Mating Factor