Because CGCFinder predicted no CGC for this PUL, the gene cluster depicted below contains dbCAN2 and CGC signature predictions for all genes in the PUL, instead of a predicted CGC.


PUL ID

PUL0231

PubMed

22502871, Microbiol Res. 2012 Sep 6;167(8):461-9. doi: 10.1016/j.micres.2012.03.004. Epub 2012 Apr 12.

Characterization method

enzyme activity assay, cosmid library screening

Genomic accession number

AY870655.1

Nucelotide position range

355-2710

Substrate

beta-glucoside

Loci

bglYK

Species

Pectobacterium carotovorum subsp. carotovorum/555

Degradation or Biosynthesis

degradation

Gene Name

Locus Tag

Protein ID

Gene Position

GenBank Contig Range

EC Number

bglY - AAX76617.1 355 - 1780 (+) AY870655.1:710-2135 -
bglK - AAX76618.1 1873 - 2710 (+) AY870655.1:2228-3065 -

Cluster number

0

Gene name

Gene position

Gene type

Found by CGCFinder?

- 356 - 1780 (+) CAZyme: GH1 No
- 1874 - 2710 (+) CDS No

PUL ID

PUL0231

PubMed

22502871, Microbiol Res. 2012 Sep 6;167(8):461-9. doi: 10.1016/j.micres.2012.03.004. Epub 2012 Apr 12.

Title

Cloning and biochemical analysis of beta-glucoside utilization (bgl) operon without phosphotransferase system in Pectobacterium carotovorum subsp. carotovorum LY34.

Author

An CL, Kim MK, Kang TH, Kim J, Kim H, Yun HD

Abstract

beta-Glucosidases are widespread in bacteria and involved in the metabolism of various carbohydrate substrates. Studying of beta-glucoside utilization (bgl) operons on operon of Pectobacterium carotovorum subsp. carotovorum LY34 (Pcc LY34) will help us understanding how beta-glucoside utilization (bgl) operon can cooperate with other systems in bacterium caused soft-rot disease. Pcc LY34 causes soft-rot disease in plants and expresses multiple enzymatic forms of beta-glucosidases. To fully explore the beta-glucoside utilization system in Pcc LY34, we have isolated a bgl operon from a genomic library for screening of beta-glucosidase activities. Sequence analysis of a 3050bp cloned DNA fragment (accession number AY870655) shows two open reading frames (bglY and bglK) that are predicted to encode proteins of 474 and 278 amino acid residues, respectively. Pair wise similarity analysis suggests BglY is a beta-glucosidase (a member of glycosyl hydrolase family 1) and BglK is a transcriptional antiterminator protein. bglY promoter region contains an inverted repeat sequence similar to transcriptional terminator. Different from other four beta-glucoside utilization operons of Pcc LY34 strain, BglY contains signal peptide sequences as extracellular beta-glucosidase. Comparisons of five beta-glucoside utilization operons of Pcc LY34 strain showed that bglYK operon does not have phosphotransferase system domain which are responsible for sugar transportation. BglY shares 33-44% identity with other four beta-glucosidases of Pcc LY34 strain. Enzyme assay showed that purified BglY enzyme hydrolyzed salicin, arbutin, pNPG, and MUG, and exhibited maximal activity at pH 7.0 and 40 degrees C. This activity was enhanced Mg(2+). Site-directed mutagenesis revealed E166 and E371 are critical of BglY's beta-glucosidase activity.