Scaled-up expression of human alpha 2,6(N)sialyltransferase in Saccharomyces cerevisiae.
Expression of recombinant full length human alpha 2,6(N)sialyltransferase has been scaled-up in S. cerevisiae in a 150-l bioreactor yielding 47 U at a concentration of 0.31 U/l. The protein specific activity as measured in reconstituted yeast lyophilisate was 0.8 mU/mg protein. The recombinant enzyme exhibited similar Michaelis constants as previously determined for the native rat enzyme. By immunoblotting the enzyme was shown to be heterogeneous by size (44-48 kD) and N-glycosylated. We conclude that recombinant alpha 2,6(N)sialyltransferase expressed in S. cerevisiae is retained in the endoplasmic reticulum as a fully active enzyme.
Borsig
L
L
Institute of Physiology, University of Zurich, Switzerland.
Ivanov
S X
SX
Herrmann
G F
GF
Kragl
U
U
Wandrey
C
C
Berger
E G
EG
D015153:Blotting, Western; D006801:Humans; D007700:Kinetics; D008970:Molecular Weight; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D012799:Sialyltransferases / Q000096:biosynthesis* / Q000378:metabolism
D011994:Recombinant Proteins; D012799:Sialyltransferases