[Engineering of a System for the Production of Mutant Human Alpha-Fetoprotein in the Methylotrophic Yeast Pichia pastoris].
A system for the production of mutant recombinant human alpha-fetoprotein (rhAFPO) lacking the glycosylation site has been engineered in the yeast Pichia pastoris. A strain of the methylotrophic yeast Pichia pastoris GS 115/pPICZ?A/rhAFP0, which produces unglycosylated rhAFPO and secretes it to the culture medium, has been constructed. Optimization and scale-up of the fermentation technology have resulted in an increase in the rhAFP0 yield to 20 mg/L. A scheme of isolation and purification of biologically active rhAFP0 has been developed. The synthesized protein has the antitumor activity, which is analogous to the activity of natural human embryonic alpha-fetoprotein.
E V
EV
Morozkina
E A
EA
Vavilova
S S
SS
Zatsepin
E V
EV
Klyachko
T A
TA
Yagudin
A M
AM
Chulkin
I V
IV
Dudich
L N
LN
Semenkova
I V
IV
Churilova
S V
SV
Benevolenskii
D045744:Cell Line, Tumor; D005285:Fermentation; D006801:Humans; D050505:Mutant Proteins / Q000008:administration & dosage / Q000096:biosynthesis* / Q000235:genetics; D009369:Neoplasms / Q000188:drug therapy*; D010843:Pichia / Q000235:genetics; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D012441:Saccharomyces cerevisiae / Q000235:genetics; D000509:alpha-Fetoproteins / Q000008:administration & dosage / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification
D050505:Mutant Proteins; D011994:Recombinant Proteins; D000509:alpha-Fetoproteins