A New Regulatory Locus of the Maltose Regulon in Klebsiella pneumoniae Strain K21 Identified by the Study of Pullulanase Secretion Mutants

Abstract

This study has shown that Klebsiella pneumoniae strain K21 differs from the previously characterized and closely related K. pneumoniae strain PAP996 in that expression of the pullulanase gene (pulA) and other genes of the maltose regulon is partially independent of exogenous inducer (maltose/maltotriose). Mutants of strain K21 which are defective in pullulanase synthesis and/or secretion were isolated following Tn10 mutangenesis. Three phenotypic classes of mutants were identified. Class I mutants were defective in the surface localization and secretion of pullulanase. Class II mutants did not secrete detectable levels of pullulanase but were able to export pullunase to the cell surface. Class II mutants also expressed pullanase and other maltose-regulated genes at markedly lower levels than those found in the parent strain under non-inducing conditions. The single class IH mutant was intermediate between K21 and class I mutants; most of the cell-associated pullulanase was localized at the cell surface whilst a significant amount was secreted into the medium. Mapping indicated that all but three of the Tn10 insertions were adjacent to, and at either side of, pulA. One class II mutant carried a Tn10 insertion in or close to malT whereas in the remaining class II mutants the insertions were located at least 4 kb upstream of pulA in a region which may define a new regulatory locus of the maltose operon.