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PLANT PHYSIOLOGY , Vol 104, Issue 4 1449-1453, Copyright © 1994 by American Society of Plant Biologists
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METABOLISM AND ENZYMOLOGY |
Expression of Branching Enzyme I of Maize Endosperm in Escherichia coli
H. P. Guan, T. Baba and J. Preiss
Department of Biochemistry, Michigan State University, East Lansing, Michigan 48824 (H.P.G., J.P.)
The gene encoding for mature branching enzyme (BE) I (BEI) of maize (Zea
mays L.) endosperm has been expressed in Escherichia coli using the T7
promoter. The expressed BEI was purified to near homogeneity so that
amylolytic activity and bacterial BE could be completely eliminated from
the BE preparation. The recombinant enzyme showed properties very similar
to those of BEI purified from developing maize endosperm with respect to
branching amylose and amylopectin. This result confirmed our earlier report
that maize endosperm BEI had a higher rate of branching amylose and a much
lower rate (less than 10% of that of branching amylose) of branching
amylopectin. This study also showed a great advantage in purifying BEI from
the bacterial expression system rather than from developing maize
endosperm. Most important, this study has established the system with which
to study the structure-function relationships of the maize BEI using
site-directed mutagenesis.
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