1887

Abstract

The EnzymeII of the phosphoenolpyruvate- (PEP-) dependent phosphotransferase system catalyses the uptake and concomitant phosphorylation of -glucosides by ; it is specified by the gene . The nucleotide sequence of a 3·6 kb dIII restriction fragment spanning , cloned on a plasmid, was determined. DNA analysis strongly suggests that the published order of this and other genes involved in -glucoside utilization, , is incorrect, and that the regulatory gene may be located upstream of the structural genes and . From the deduced amino acid sequence it is predicted that the membrane protein specified by consists of 625 amino acid residues (66·48 kDa). The protein has the hydropathic profile expected of an integral membrane protein (average hydropathy = 0·62). Comparisons between the amino acid sequences deduced for the EnzymeII and for the mannitol-specific EnzymeII show that these proteins are related, and a little direct homology is apparent. A 2·3 kb I fragment spanning was subcloned into an expression vector which carries the λp promoter and then transformed into a host strain which produces thermolabile c1857 repressor and the anti-terminator N; thermoinduction resulted in the overproduction of a membrane protein and the appearance of Bgl activity.

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1987-03-01
2021-07-31
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