A functional analysis of the Bifidobacterium longum cscA and scrP genes in sucrose utilization.
The role of genes involved in sucrose catabolism was investigated with a view to designing effective prebiotic substrates to encourage the growth of Bifidobacterium in the gut. Two gene clusters coding for sucrose utilisation in Bifidobacterium longum NCC2705 were identified in the published genome. The genes encoding putative sucrose degrading enzymes, namely, the scrP (sucrose phosphorylase) and the cscA (beta-fructofuranosidase), were cloned from B. longum NCIMB 702259(T) and expressed in Escherichia coli DH5alpha. Both complemented the sucrase negative phenotype of untransformed cells and showed specific sucrase activity. Transcriptional analysis of the expression of the genes in B. longum grown in the presence of various carbohydrate substrates showed induction of scrP gene expression in the presence of sucrose and raffinose, but not in the presence of glucose. The cscA gene showed no increased transcription in B. longum grown in the presence of any of the carbohydrates tested. Phylogenetic analysis indicates that the B. longum CscA protein belongs to a distinct phylogenetic cluster of intracellular fructosidases, which specifically cleave the shorter fructose oligosaccharides.
Kullin
B
B
Department of Molecular and Cell Biology, University of Cape Town, Cape Town, South Africa.
Abratt
V R
VR
Reid
S J
SJ
D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D001644:Bifidobacterium / Q000235:genetics* / Q000378:metabolism*; D003470:Culture Media / Q000737:chemistry; D015964:Gene Expression Regulation, Bacterial; D010802:Phylogeny; D012329:RNA, Bacterial / Q000378:metabolism; D013395:Sucrose / Q000032:analysis / Q000378:metabolism*; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D003470:Culture Media; D012329:RNA, Bacterial; D013395:Sucrose