Biochemical evidence that starch breakdown by Bacteroides thetaiotaomicron involves outer membrane starch-binding sites and periplasmic starch-degrading enzymes.
Bacteroides thetaiotaomicron can utilize amylose, amylopectin, and pullulan as sole sources of carbon and energy. The enzymes that degrade these polysaccharides were found to be primarily cell associated rather than extracellular. Although some activity was detected in extracellular fluid, this appeared to be the result of cell lysis. The cell-associated amylase, amylopectinase, and pullulanase activities partitioned similarly to the periplasmic marker, acid phosphatase, when cells were exposed to a cold-shock treatment. Two other enzymes associated with starch breakdown, alpha-glucosidase and maltase, appeared to be located in the cytoplasm. Intact cells of B. thetaiotaomicron were found to bind 14C-starch. Binding was probably mediated by a protein because it was saturable and was decreased by treatment of cells with proteinase K. Results of competition experiments showed that the starch-binding proteins had a preference for maltodextrins larger than maltohexaose and a low affinity for maltose and maltotriose. Both the degradative enzymes and starch binding were induced by maltose. These findings indicate that starch utilization by B. thetaiotaomicron apparently does not involve secretion of extracellular enzymes. Rather, binding of the starch molecule to the cell surface appears to be a first step to passing the molecule through the outer membrane and into the periplasmic space.
Anderson
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KL
Department of Microbiology, University of Illinois, Urbana 61801.
Salyers
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AA
D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism; D001439:Bacteroides / Q000378:metabolism* / Q000648:ultrastructure; D001665:Binding Sites; D001667:Binding, Competitive; D002451:Cell Compartmentation; D002462:Cell Membrane / Q000378:metabolism; D003470:Culture Media; D008320:Maltose / Q000494:pharmacology; D008970:Molecular Weight; D013213:Starch / Q000378:metabolism*; D013499:Surface Properties
D001425:Bacterial Outer Membrane Proteins; D003470:Culture Media; D008320:Maltose; D013213:Starch