Genetic evidence that outer membrane binding of starch is required for starch utilization by Bacteroides thetaiotaomicron.
Mutagenesis of Bacteroides thetaiotaomicron with the transposon Tn4351 produced five classes of mutants that were not able to grow on amylose or amylopectin. These classes of mutants differed in their ability to grow on maltoheptaose (G7) and in the level of starch-degrading enzymes produced when bacteria were grown on maltose. All of the mutants were deficient in starch binding. Since one class of mutants retained normal levels of starch-degrading enzymes, this indicates that binding of the starch molecule by a cell surface receptor is necessary for starch utilization by B. thetaiotaomicron. Analysis of a starch-negative mutant that grew on G7 indicated that B. thetaiotaomicron possessed two starch-binding components or sites. One component (site A), apparently missing in this mutant, had an absolute preference for larger starch oligomers, whereas the other component (site M) also had a high affinity for maltodextrins (G4 through G7). Mutants not able to grow on maltodextrins (greater than G4) probably lacked both of these binding components. Only one class of mutants did not grow normally on maltose, but instead had a 4- to 5-h lag on maltose and a slower growth rate than the wild type. This class of mutants did not produce any of the starch-degrading enzymes or bind starch, even when growing on maltose. Such a phenotype probably resulted from transposon inactivation of a central regulatory gene or a gene encoding an enzyme that produces the inducer. The fact that both the degradative enzymes and the starch-binding activity were affected in this mutant indicates that genes encoding the cell surface starch-binding site are under the same regulatory control as genes encoding the enzymes.
Anderson
K L
KL
Department of Microbiology, University of Illinois, Urbana 61801.
Salyers
A A
AA
D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism; D001439:Bacteroides / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D001692:Biological Transport; D015139:Blotting, Southern; D002451:Cell Compartmentation; D002462:Cell Membrane / Q000378:metabolism; D004269:DNA, Bacterial / Q000235:genetics; D008320:Maltose / Q000378:metabolism; D009154:Mutation; D013213:Starch / Q000378:metabolism*; D013329:Structure-Activity Relationship
D001425:Bacterial Outer Membrane Proteins; D004269:DNA, Bacterial; D008320:Maltose; D013213:Starch