Substrate-driven gene expression in Roseburia inulinivorans

Importance of inducible enzymes in the utilization of inulin and starch

Karen P. Scott, Jennifer C. Martin, Christophe Chassard, Marlene Clerget, Joanna Potrykus, Gillian P. Campbell, Claus-Dieter Mayer, Pauline Young, Garry Rucklidge, Alan G. Ramsay, Harry J. Flint

Research output: Contribution to journalArticle

60 Citations (Scopus)

Abstract

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 beta-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 beta-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 beta-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 a-amylase and two distinct alpha-glucanotransferases together with a gene encoding a flagellin protein. The latter response may be concerned with improving bacterial access to insoluble starch particles.

Original languageEnglish
Pages (from-to)4672-4679
Number of pages8
JournalPNAS
Volume108
Issue numberSupplement 1
DOIs
Publication statusPublished - 15 Mar 2011

Keywords

  • anaerobic gut bacteria
  • differential gene expression
  • fructo-oligosaccharides
  • butyrate
  • prebiotic
  • human fecal flora
  • beta-fructofuranosidase
  • human gut
  • human colon
  • lactobacillus-acidophilus
  • bifidobacterium-lactis
  • molecular analysis
  • escherichia-coli
  • dominant group
  • bacteria

Cite this

Substrate-driven gene expression in Roseburia inulinivorans : Importance of inducible enzymes in the utilization of inulin and starch. / Scott, Karen P.; Martin, Jennifer C.; Chassard, Christophe; Clerget, Marlene; Potrykus, Joanna; Campbell, Gillian P.; Mayer, Claus-Dieter; Young, Pauline; Rucklidge, Garry; Ramsay, Alan G.; Flint, Harry J.

In: PNAS, Vol. 108, No. Supplement 1, 15.03.2011, p. 4672-4679.

Research output: Contribution to journalArticle

Scott, KP, Martin, JC, Chassard, C, Clerget, M, Potrykus, J, Campbell, GP, Mayer, C-D, Young, P, Rucklidge, G, Ramsay, AG & Flint, HJ 2011, 'Substrate-driven gene expression in Roseburia inulinivorans: Importance of inducible enzymes in the utilization of inulin and starch', PNAS, vol. 108, no. Supplement 1, pp. 4672-4679. https://doi.org/10.1073/pnas.1000091107
Scott, Karen P. ; Martin, Jennifer C. ; Chassard, Christophe ; Clerget, Marlene ; Potrykus, Joanna ; Campbell, Gillian P. ; Mayer, Claus-Dieter ; Young, Pauline ; Rucklidge, Garry ; Ramsay, Alan G. ; Flint, Harry J. / Substrate-driven gene expression in Roseburia inulinivorans : Importance of inducible enzymes in the utilization of inulin and starch. In: PNAS. 2011 ; Vol. 108, No. Supplement 1. pp. 4672-4679.
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abstract = "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 beta-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 beta-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 beta-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 a-amylase and two distinct alpha-glucanotransferases together with a gene encoding a flagellin protein. The latter response may be concerned with improving bacterial access to insoluble starch particles.",
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T1 - Substrate-driven gene expression in Roseburia inulinivorans

T2 - Importance of inducible enzymes in the utilization of inulin and starch

AU - Scott, Karen P.

AU - Martin, Jennifer C.

AU - Chassard, Christophe

AU - Clerget, Marlene

AU - Potrykus, Joanna

AU - Campbell, Gillian P.

AU - Mayer, Claus-Dieter

AU - Young, Pauline

AU - Rucklidge, Garry

AU - Ramsay, Alan G.

AU - Flint, Harry J.

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N2 - 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 beta-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 beta-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 beta-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 a-amylase and two distinct alpha-glucanotransferases together with a gene encoding a flagellin protein. The latter response may be concerned with improving bacterial access to insoluble starch particles.

AB - 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 beta-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 beta-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 beta-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 a-amylase and two distinct alpha-glucanotransferases together with a gene encoding a flagellin protein. The latter response may be concerned with improving bacterial access to insoluble starch particles.

KW - anaerobic gut bacteria

KW - differential gene expression

KW - fructo-oligosaccharides

KW - butyrate

KW - prebiotic

KW - human fecal flora

KW - beta-fructofuranosidase

KW - human gut

KW - human colon

KW - lactobacillus-acidophilus

KW - bifidobacterium-lactis

KW - molecular analysis

KW - escherichia-coli

KW - dominant group

KW - bacteria

U2 - 10.1073/pnas.1000091107

DO - 10.1073/pnas.1000091107

M3 - Article

VL - 108

SP - 4672

EP - 4679

JO - PNAS

JF - PNAS

SN - 0027-8424

IS - Supplement 1

ER -