PUL ID

PUL0394

PubMed

20679207, Proc Natl Acad Sci U S A. 2011 Mar 15;108 Suppl 1:4672-9. doi: 10.1073/pnas.1000091107. Epub 2010 Aug 2.

Characterization method

microarray

Genomic accession number

GU591787.1

Nucelotide position range

20-7392

Substrate

fructan,inulin

Loci

ADF57363.1-ADF57368.1

Species

Roseburia inulinivorans/360807

Degradation or Biosynthesis

degradation

Gene Name

Locus Tag

Protein ID

Gene Position

GenBank Contig Range

EC Number

- - ADF57363.1 20 - 1046 (-) GU591787.1:40-1066 -
- - ADF57364.1 1302 - 2241 (+) GU591787.1:1322-2261 -
- - ADF57365.1 2253 - 3150 (+) GU591787.1:2273-3170 -
- - ADF57366.1 3169 - 4840 (+) GU591787.1:3189-4860 -
- - ADF57367.1 4856 - 6374 (+) GU591787.1:4876-6394 -
6-pfk - ADF57368.1 6435 - 7392 (+) GU591787.1:6455-7412 -

Cluster number

1

Gene name

Gene position

Gene type

Found by CGCFinder?

- 21 - 1046 (-) TF: DBD-Pfam|LacI,DBD-SUPERFAMILY|0036955 No
- 1303 - 2241 (+) TC: gnl|TC-DB|Q9KWT8|3.A.1.1.10 Yes
- 2254 - 3150 (+) TC: gnl|TC-DB|Q9WYR1|3.A.1.1.39 Yes
- 3170 - 4840 (+) other Yes
- 4857 - 6374 (+) CAZyme: GH32 Yes
- 6436 - 7392 (+) STP: STP|PfkB No

PUL ID

PUL0394

PubMed

20679207, Proc Natl Acad Sci U S A. 2011 Mar 15;108 Suppl 1:4672-9. doi: 10.1073/pnas.1000091107. Epub 2010 Aug 2.

Title

Substrate-driven gene expression in Roseburia inulinivorans: importance of inducible enzymes in the utilization of inulin and starch.

Author

Scott KP, Martin JC, Chassard C, Clerget M, Potrykus J, Campbell G, Mayer CD, Young P, Rucklidge G, Ramsay AG, Flint HJ

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 alpha-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.