Cellodextrins, the incomplete hydrolysis products from insoluble cellulose, are accessible as a carbon source
to certain members of the human gut microbiota, such as Bifidobacterium breve UCC2003. Transcription of the
cldEFGC gene cluster of B. breve UCC2003 was shown to be induced upon growth on cellodextrins, implicating
this cluster in the metabolism of these sugars. Phenotypic analysis of a B. breve UCC2003::cldE insertion
mutant confirmed that the cld gene cluster is exclusively required for cellodextrin utilization by this commensal.
Moreover, our results suggest that transcription of the cld cluster is controlled by a LacI-type regulator
encoded by cldR, located immediately upstream of cldE. Gel mobility shift assays using purified CldRHis
(produced by the incorporation of a His12-encoding sequence into the 3 end of the cldC gene) indicate that the
cldEFGC promoter is subject to negative control by CldRHis, which binds to two inverted repeats. Analysis by
high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) of
medium samples obtained during growth of B. breve UCC2003 on a mixture of cellodextrins revealed its ability
to utilize cellobiose, cellotriose, cellotetraose, and cellopentaose, with cellotriose apparently representing the
preferred substrate. The cldC gene of the cld operon of B. breve UCC2003 is, to the best of our knowledge, the
first described bifidobacterial -glucosidase exhibiting hydrolytic activity toward various cellodextrins.
Cellodextrins, the incomplete hydrolysis products from insoluble cellulose, are accessible as a carbon source
to certain members of the human gut microbiota, such as Bifidobacterium breve UCC2003. Transcription of the
cldEFGC gene cluster of B. breve UCC2003 was shown to be induced upon growth on cellodextrins, implicating
this cluster in the metabolism of these sugars. Phenotypic analysis of a B. breve UCC2003::cldE insertion
mutant confirmed that the cld gene cluster is exclusively required for cellodextrin utilization by this commensal.
Moreover, our results suggest that transcription of the cld cluster is controlled by a LacI-type regulator
encoded by cldR, located immediately upstream of cldE. Gel mobility shift assays using purified CldRHis
(produced by the incorporation of a His12-encoding sequence into the 3 end of the cldC gene) indicate that the
cldEFGC promoter is subject to negative control by CldRHis, which binds to two inverted repeats. Analysis by
high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) of
medium samples obtained during growth of B. breve UCC2003 on a mixture of cellodextrins revealed its ability
to utilize cellobiose, cellotriose, cellotetraose, and cellopentaose, with cellotriose apparently representing the
preferred substrate. The cldC gene of the cld operon of B. breve UCC2003 is, to the best of our knowledge, the
first described bifidobacterial -glucosidase exhibiting hydrolytic activity toward various cellodextrins.
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