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Cite: NAR/gkaa742 2020



PUL General Information

Literature Information

PUL ID

PUL0455

PubMed

12513973, Appl Environ Microbiol. 2003 Jan;69(1):24-32. doi: 10.1128/AEM.69.1.24-32.2003.

Characterization method

clone and expression,genes induced in presence of substrate,enzyme activity assay

Genomic accession number

AF441242.1

Nucelotide position range

771-5247

Substrate

sucrose

Loci

scrT-scrP-scrR

Species

Bifidobacterium animalis/28025

Degradation or Biosynthesis

degradation

Gene Name

Locus Tag

Protein ID

Gene Position

GenBank Contig Range

EC Number

scrT - AAN01604.1 770 - 2372 (-) AF441242.1:1541-3143 -
scrP - AAN01605.1 2440 - 3961 (-) AF441242.1:3211-4732 -
scrR - AAN01606.1 4266 - 5247 (+) AF441242.1:5037-6018 -

Cluster number

1

Gene name

Gene position

Gene type

Found by CGCFinder?

- 771 - 2072 (-) TC: gnl|TC-DB|J9W1F3|2.A.1.2.93 Yes
- 2441 - 3961 (-) CAZyme: GH13|GH13_18 Yes
- 4267 - 5247 (+) TF: DBD-Pfam|LacI,DBD-SUPERFAMILY|0036955 No

PUL ID

PUL0455

PubMed

12513973, Appl Environ Microbiol. 2003 Jan;69(1):24-32. doi: 10.1128/AEM.69.1.24-32.2003.

Title

Induction of sucrose utilization genes from Bifidobacterium lactis by sucrose and raffinose.

Author

Trindade MI, Abratt VR, Reid SJ

Abstract

The probiotic organism Bifidobacterium lactis was isolated from a yoghurt starter culture with the aim of analyzing its use of carbohydrates for the development of prebiotics. A sucrose utilization gene cluster of B. lactis was identified by complementation of a gene library in Escherichia coli. Three genes, encoding a sucrose phosphorylase (ScrP), a GalR-LacI-type transcriptional regulator (ScrR), and a sucrose transporter (ScrT), were identified by sequence analysis. The scrP gene was expressed constitutively from its own promoter in E. coli grown in complete medium, and the strain hydrolyzed sucrose in a reaction that was dependent on the presence of phosphates. Primer extension experiments with scrP performed by using RNA isolated from B. lactis identified the transcriptional start site 102 bp upstream of the ATG start codon, immediately adjacent to a palindromic sequence resembling a regulator binding site. In B. lactis, total sucrase activity was induced by the presence of sucrose, raffinose, or oligofructose in the culture medium and was repressed by glucose. RNA analysis of the scrP, scrR, and scrT genes in B. lactis indicated that expression of these genes was influenced by transcriptional regulation and that all three genes were similarly induced by sucrose and raffinose and repressed by glucose. Analysis of the sucrase activities of deletion constructs in heterologous E. coli indicated that ScrR functions as a positive regulator.