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Substrate-driven gene expression in Roseburia inulinivorans : Importance of inducible enzymes in the utilization of inulin and starch
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Substrate-driven gene expression in Roseburia inulinivorans : Importance of inducible enzymes in the utilization of inulin and starch
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Journal Article
Substrate-driven gene expression in Roseburia inulinivorans : Importance of inducible enzymes in the utilization of inulin and starch
2011
Overview
Roseburia inulinivorans is a recently identified motile representative of the Firmicutes that contributes to butyrate formation from a variety of dietary polysaccharide substrates in the human large intestine. Microarray analysis was used here to investigate substrate-driven gene-expression changes in R. inulinivorans A2-194. A cluster of fructo-oligosaccharide/inulin utilization genes induced during growth on inulin included one encoding a β-fructofuranosidase protein that was prominent in the proteome of inulin-grown cells. This cluster also included a 6-phosphofructokinase and an ABC transport system, whereas a distinct inulin-induced 1-phosphofructokinase was linked to a fructose-specific phosphoenolpyruvate-dependent sugar phosphotransferase system (PTS II transport enzyme). Real-time PCR analysis showed that the β-fructofuranosidase and adjacent ABC transport protein showed greatest induction during growth on inulin, whereas the 1-phosphofructokinase enzyme and linked sugar phosphotransferase transport system were most strongly up-regulated during growth on fructose, indicating that these two clusters play distinct roles in the use of inulin. The R. inulinivorans β-fructofuranosidase was overexpressed in Escherichia coli and shown to hydrolyze fructans ranging from inulin down to sucrose, with greatest activity on fructo-oligosaccharides. Genes induced on starch included the major extracellular α-amylase and two distinct α-glucanotransferases together with a gene encoding a flagellin protein. The latter response may be concerned with improving bacterial access to insoluble starch particles.
Publisher
National Academy of Sciences,HAL CCSD,National Acad Sciences
Subject
/ ATP binding cassette transporters
/ ATP-Binding Cassette Transporters - metabolism
/ Bacteria
/ beta-Fructofuranosidase - metabolism
/ Culture Media - pharmacology
/ Enzymes
/ Fructans
/ fructose
/ Gene Expression Regulation, Bacterial
/ Genes
/ Gram-Positive Endospore-Forming Rods - enzymology
/ Gram-Positive Endospore-Forming Rods - genetics
/ Humans
/ Intestine, Large - microbiology
/ inulin
/ Oligonucleotide Array Sequence Analysis
/ Phosphofructokinase-1 - metabolism
/ proteome
/ quantitative polymerase chain reaction
/ Reverse Transcriptase Polymerase Chain Reaction
/ starch
/ Starches
/ sucrose
