Cloning and expression of the multiple sugar metabolism (msm) operon of Streptococcus mutans in heterologous streptococcal hosts

The multiple sugar metabolism (msm) operon of Streptococcus mutans is responsible for the uptake and metabolism of a variety of sugars. In order to further characterize the substrate specificities of the transport system, a 12-kb region of DNA containing the entire msm operon was cloned, via a novel two-step integration strategy, into the chromosomes of two heterologous streptococcal strains, Streptococcus gordonii Challis and Streptococcus anginosus Is57, as well as the chromosome of a natural isolate of S. mutans with a deletion of the msm region. These strains are unable to transport or ferment melibiose, raffinose, or isomaltosaccharides, but the newly constructed recombinants gained the ability to ferment all of these sugars. The S. gordonii Challis construct containing msm was shown to transport radiolabelled melibiose, raffinose, isomaltotriose, and isomaltotetraose, and the transport function was also subjected to induction by raffinose, an inducer of the msm operon in S. mutans. The results confirm the role of the msm operon in the transport and metabolism of melibiose, raffinose, and isomaltosaccharides.

[1]  J. Ferretti,et al.  A binding protein-dependent transport system in Streptococcus mutans responsible for multiple sugar metabolism. , 1992, The Journal of biological chemistry.

[2]  D. Beighton,et al.  A scheme for the identification of viridans streptococci. , 1991, Journal of medical microbiology.

[3]  R. Russell,et al.  Chromosomal Deletions in Melibiose-negative Isolates of Streptococcus mutans , 1991, Journal of dental research.

[4]  G. Dunny,et al.  Improved electroporation and cloning vector system for gram-positive bacteria , 1991, Applied and environmental microbiology.

[5]  J. Ferretti,et al.  Nucleotide sequence of the dextran glucosidase (dexB) gene of Streptococcus mutans. , 1990, Journal of general microbiology.

[6]  S. Dashper,et al.  Characterization of transmembrane movement of glucose and glucose analogs in Streptococcus mutants Ingbritt , 1990, Journal of bacteriology.

[7]  J. Lodge,et al.  Carbohydrate uptake in the oral pathogen Streptococcus mutans: mechanisms and regulation by protein phosphorylation. , 1989, Biochimie.

[8]  J. Claverys,et al.  The difficulty of cloning Streptococcus pneumoniae mal and ami loci in Escherichia coli: toxicity of malX and amiA gene products. , 1989, Gene.

[9]  J. Ferretti,et al.  Streptococcus mutans gtfA gene specifies sucrose phosphorylase , 1988, Infection and immunity.

[10]  P. Hanawalt,et al.  High-efficiency transformation of bacterial cells by electroporation , 1988, Journal of bacteriology.

[11]  J. Ferretti,et al.  Sequence analysis of the glucosyltransferase A gene (gtfA) from Streptococcus mutans Ingbritt , 1988, Infection and immunity.

[12]  W. Bowen,et al.  Tight Genetic Linkage of a Glucosyltransferase and Dextranase of Streptococcus mutans GS-5 , 1986, Journal of dental research.

[13]  I. J. Evans,et al.  A family of related ATP-binding subunits coupled to many distinct biological processes in bacteria , 1986, Nature.

[14]  M. Hofnung,et al.  Sequences of the malE gene and of its product, the maltose-binding protein of Escherichia coli K12. , 1984, The Journal of biological chemistry.

[15]  P. Marsh,et al.  Evidence that glucose and sucrose uptake in oral streptococcal bacteria involves independent phosphotransferase and proton-motive force-mediated mechanisms. , 1984, Archives of oral biology.

[16]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[17]  R. P. Evans,et al.  Novel shuttle plasmid vehicles for Escherichia-Streptococcus transgeneric cloning. , 1983, Gene.

[18]  C. Douglas,et al.  Fructosyltransferase activity of a glucan-binding protein from Streptococcus mutans. , 1983, Journal of general microbiology.

[19]  A. Slee,et al.  Sucrose transport by Streptococcus mutans. Evidence for multiple transport systems. , 1982, Biochimica et biophysica acta.

[20]  I. R. Hamilton,et al.  Evidence for the Involvement of Proton Motive Force in the Transport of Glucose by a Mutant of Streptococcus mutans Strain DR0001 Defective in Glucose-Phosphoenolpyruvate Phosphotransferase Activity , 1982, Infection and immunity.

[21]  H. Kuramitsu,et al.  Genetic Transformation of Streptococcus mutans , 1981, Infection and immunity.

[22]  S. Hamada,et al.  Biology, immunology, and cariogenicity of Streptococcus mutans. , 1980, Microbiological reviews.

[23]  A. Slee,et al.  Effect of growth conditions on sucrose phosphotransferase activity of Streptococcus mutans , 1980, Infection and immunity.

[24]  J.,et al.  Biochemical and genetic analysis of Streptococcus mutans a-galactosidase , 2022 .