[Expression Of DNA-Encoded Antidote to Organophosphorus Toxins in the Methylotrophic Yeast Pichia Pastoris].
A platform for the cloning and expression of active human butyrylcholinesterase (BuChE) in the yeast Pichia pastoris is first presented. Genetic constructs for BuChE gene expression, separately and in conjunction with a proline-rich peptide called proline-rich attachment domain (PRAD), are based on the vector pPICZαA. It is shown that the highest level of production is achieved in the expression of a BuChE gene without PRAD pPICZαA. It is found that one can obtain up to 125 mg of active enzyme from 1 L of culture medium at an optimal pH environment (pH 7.6), an optical seed culture density of 3 o.u., and an optimum methanol addition mode of (0.5% methanol in the first day and 0.2% thereafter from the second day).
S S
SS
Terekhov
T V
TV
Bobik
Yu A
YA
Mokrushina
A V
AV
Stepanova
N M
NM
Aleksandrova
I V
IV
Smirnov
A A
AA
Belogurov
N A
NA
Ponomarenko
A G
AG
Gabibov
D000931:Antidotes; D002091:Butyrylcholinesterase / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D003001:Cloning, Molecular; D004247:DNA / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D010755:Organophosphates / Q000737:chemistry / Q000633:toxicity; D010455:Peptides / Q000737:chemistry; D010843:Pichia / Q000201:enzymology / Q000235:genetics; D011392:Proline / Q000737:chemistry
D000931:Antidotes; D010755:Organophosphates; D010455:Peptides; D004247:DNA; D011392:Proline; D002091:Butyrylcholinesterase