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Cite: NAR/gkaa742 2020



PUL General Information

Literature Information

PUL ID

PUL0369

PubMed

19416976, J Biol Chem. 2009 Jul 3;284(27):18059-69. doi: 10.1074/jbc.M109.005660. Epub 2009 May 5.

Characterization method

RT-PCR

Genomic accession number

NC_004368.1

Nucelotide position range

1953787-1964447

Substrate

unsaturated hyaluronate disaccharide,chondroitin disaccharide

Loci

GBS_RS09830-GBS_RS09880

Species

Streptococcus agalactiae NEM316/211110

Degradation or Biosynthesis

degradation

Gene Name

Locus Tag

Protein ID

Gene Position

GenBank Contig Range

EC Number

- GBS_RS09830 WP_000147951.1 0 - 816 (-) NC_004368.1:1953787-1954603 -
- GBS_RS09835 WP_000355267.1 802 - 1669 (-) NC_004368.1:1954589-1955456 -
- GBS_RS09840 WP_000718802.1 1704 - 2199 (-) NC_004368.1:1955491-1955986 -
- GBS_RS09845 WP_000975716.1 2253 - 3450 (-) NC_004368.1:1956040-1957237 -
- GBS_RS09850 WP_000590444.1 3452 - 3887 (-) NC_004368.1:1957239-1957674 -
dhuD GBS_RS09855 WP_000139169.1 4164 - 4977 (+) NC_004368.1:1957951-1958764 1.1.1.127
dhuI GBS_RS09860 WP_000684791.1 4993 - 5632 (+) NC_004368.1:1958780-1959419 5.3.1.17
- GBS_RS09865 WP_000034943.1 5657 - 6665 (+) NC_004368.1:1959444-1960452 -
- GBS_RS09870 WP_000597408.1 6676 - 7315 (+) NC_004368.1:1960463-1961102 -
- GBS_RS09875 WP_000440333.1 8514 - 9876 (+) NC_004368.1:1962301-1963663 -
- GBS_RS09880 WP_000008878.1 10046 - 10661 (-) NC_004368.1:1963833-1964448 -

Cluster number

1

Gene name

Gene position

Gene type

Found by CGCFinder?

- 1 - 816 (-) TC: gnl|TC-DB|Q8DR74|4.A.6.1.14 Yes
- 803 - 1669 (-) TC: gnl|TC-DB|Q8DR75|4.A.6.1.14 Yes
- 1705 - 2199 (-) TC: gnl|TC-DB|Q8DR76|4.A.6.1.14 Yes
- 2254 - 3450 (-) CAZyme: GH88 Yes
- 3453 - 3887 (-) TC: gnl|TC-DB|Q8DR78|4.A.6.1.14 Yes
dhuD 4165 - 4977 (+) CDS No
dhuI 4994 - 5632 (+) CDS No
- 5658 - 6665 (+) STP: STP|PfkB No
- 6677 - 7315 (+) CDS No
- 8515 - 9876 (+) CDS No
- 10047 - 10661 (-) STP: STP|Nitroreductase No

PUL ID

PUL0369

PubMed

19416976, J Biol Chem. 2009 Jul 3;284(27):18059-69. doi: 10.1074/jbc.M109.005660. Epub 2009 May 5.

Title

Substrate specificity of streptococcal unsaturated glucuronyl hydrolases for sulfated glycosaminoglycan.

Author

Maruyama Y, Nakamichi Y, Itoh T, Mikami B, Hashimoto W, Murata K

Abstract

Unsaturated glucuronyl hydrolase (UGL) categorized into the glycoside hydrolase family 88 catalyzes the hydrolytic release of an unsaturated glucuronic acid from glycosaminoglycan disaccharides, which are produced from mammalian extracellular matrices through the beta-elimination reaction of polysaccharide lyases. Here, we show enzyme characteristics of pathogenic streptococcal UGLs and structural determinants for the enzyme substrate specificity. The putative genes for UGL and phosphotransferase system for amino sugar, a component of glycosaminoglycans, are assembled into a cluster in the genome of pyogenic and hemolytic streptococci such as Streptococcus agalactiae, Streptococcus pneumoniae, and Streptococcus pyogenes, which produce extracellular hyaluronate lyase as a virulent factor. The UGLs of these three streptococci were overexpressed in Escherichia coli cells, purified, and characterized. Streptococcal UGLs degraded unsaturated hyaluronate and chondroitin disaccharides most efficiently at approximately pH 5.5 and 37 degrees C. Distinct from Bacillus sp. GL1 UGL, streptococcal UGLs preferred sulfated substrates. DNA microarray and Western blotting indicated that the enzyme was constitutively expressed in S. agalactiae cells, although the expression level increased in the presence of glycosaminoglycan. The crystal structure of S. agalactiae UGL (SagUGL) was determined at 1.75 A resolution by x-ray crystallography. SagUGL adopts alpha(6)/alpha(6)-barrel structure as a basic scaffold similar to Bacillus UGL, but the arrangement of amino acid residues in the active site differs between the two. SagUGL Arg-236 was found to be one of the residues involved in its activity for the sulfated substrate through structural comparison and site-directed mutagenesis. This is the first report on the structure and function of streptococcal UGLs.