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Molecular cloning, functional expression and purification of a glucan branching enzyme from Neisseria
V Büttcher1, M Quanz, L Willmitzer
1Institut für Genbiologische Forschung GmbH Berlin, Ihnestr. 63, D-14195, Berlin, Germany. buettcher@mpimp-golm.mpg.de
Abstract:
The nucleotide sequence containing the complete structural information for a glucan branching enzyme was isolated from a Neisseria denitrificans genomic library. The gene was expressed in Escherichia coli and the active recombinant protein was purified. The deduced protein of 762 amino acids with a calculated molecular weight of 86313 Da shows similarity to the primary protein sequences of other known glucan branching enzymes. Amino acid sequencing of the isolated protein by Edman degradation confirmed the deduced start codon of the structural gene of the glucan branching enzyme. The purified glucan branching enzyme has a stimulating effect on the Neisseria amylosucrase activity.