SCALED-UP EXPRESSION OF HUMAN ALPHA-2,6(N)SIALYLTRANSFERASE IN SACCHAROMYCES-CEREVISIAE

被引:19
作者
BORSIG, L [1 ]
IVANOV, SX [1 ]
HERRMANN, GF [1 ]
KRAGL, U [1 ]
WANDREY, C [1 ]
BERGER, EG [1 ]
机构
[1] FORSCHUNGSZENTRUM JULICH, FORSCHUNGSZENTRUM, INST BIOTECHNOL 2, D-52425 JULICH, GERMANY
关键词
D O I
10.1006/bbrc.1995.1621
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Expression of recombinant full length human alpha 2,6(N)sialyltransferase has been scaled-up in S. cerevisiae in a 150-1 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. (C) 1995 Academic Press, Inc.
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页码:14 / 20
页数:7
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