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Cite: NAR/gkaa742 2020



PUL General Information

Literature Information

PUL ID

PUL0006

PubMed

24637762, Acta Crystallogr F Struct Biol Commun. 2014 Feb;70(Pt 2):225-31. doi: 10.1107/S2053230X13034778. Epub 2014 Jan 21.

Characterization method

enzyme activity assay

Genomic accession number

KF840174.1

Nucelotide position range

236-13867

Substrate

galactan

Loci

gan1D-ganR2-gan4c-AHL67643.1-ganP-galS-galT2-galE2-galF2-galG2-frk

Species

Geobacillus stearothermophilus/1422

Degradation or Biosynthesis

degradation

Gene Name

Locus Tag

Protein ID

Gene Position

GenBank Contig Range

EC Number

gan1D - AHL67640.1 0 - 1437 (+) KF840174.1:236-1673 3.2.1.85
ganR2 - AHL67641.1 1442 - 2198 (-) KF840174.1:1678-2434 -
gan4C - AHL67642.1 2322 - 3666 (+) KF840174.1:2558-3902 3.2.1.86
- - AHL67643.1 3879 - 4746 (+) KF840174.1:4115-4982 -
ganP - AHL67644.1 4965 - 5979 (+) KF840174.1:5201-6215 -
ganS - AHL67645.1 6141 - 7611 (+) KF840174.1:6377-7847 -
ganT - AHL67646.1 7588 - 9130 (+) KF840174.1:7824-9366 -
ganE2 - AHL67647.1 9444 - 10743 (+) KF840174.1:9680-10979 -
ganF2 - AHL67648.1 10804 - 11776 (+) KF840174.1:11040-12012 -
ganG2 - AHL67649.1 11775 - 12597 (+) KF840174.1:12011-12833 -
frk - AHL67650.1 12759 - 13632 (+) KF840174.1:12995-13868 -

Cluster number

1

Gene name

Gene position

Gene type

Found by CGCFinder?

- 1 - 1437 (+) CAZyme: GH1 Yes
- 1443 - 2198 (-) STP: STP|HTH_6,STP|SIS Yes
- 2323 - 3666 (+) other Yes
- 3880 - 4746 (+) other Yes
- 4966 - 5979 (+) TC: gnl|TC-DB|G4FGN5|3.A.1.2.20 Yes
- 6142 - 7611 (+) TC: gnl|TC-DB|F4LXP4|8.A.59.2.1 Yes
- 7589 - 9130 (+) STP: STP|Response_reg,STP|HTH_18,STP|HTH_AraC Yes
- 9445 - 10743 (+) TC: gnl|TC-DB|Q72KX2|3.A.1.1.24 Yes
- 10805 - 11776 (+) TC: gnl|TC-DB|Q72KX3|3.A.1.1.24 Yes
- 11776 - 12597 (+) TC: gnl|TC-DB|Q72KX4|3.A.1.1.24 Yes
- 12760 - 13632 (+) CDS No

PUL ID

PUL0006

PubMed

24637762, Acta Crystallogr F Struct Biol Commun. 2014 Feb;70(Pt 2):225-31. doi: 10.1107/S2053230X13034778. Epub 2014 Jan 21.

Title

Purification, crystallization and preliminary crystallographic analysis of Gan1D, a GH1 6-phospho-beta-galactosidase from Geobacillus stearothermophilus T1.

Author

Lansky S, Zehavi A, Dann R, Dvir H, Belrhali H, Shoham Y, Shoham G

Abstract

Geobacillus stearothermophilus T1 is a Gram-positive thermophilic soil bacterium that contains an extensive system for the utilization of plant cell-wall polysaccharides, including xylan, arabinan and galactan. The bacterium uses a number of extracellular enzymes that break down the high-molecular-weight polysaccharides into short oligosaccharides, which enter the cell and are further hydrolyzed into sugar monomers by dedicated intracellular glycoside hydrolases. The interest in the biochemical characterization and structural analysis of these proteins originates mainly from the wide range of their potential biotechnological applications. Studying the different hemicellulolytic utilization systems in G. stearothermophilus T1, a new galactan-utilization gene cluster was recently identified, which encodes a number of proteins, one of which is a GH1 putative 6-phospho-beta-galactosidase (Gan1D). Gan1D has recently been cloned, overexpressed, purified and crystallized as part of its comprehensive structure-function study. The best crystals obtained for this enzyme belonged to the triclinic space group P1, with average crystallographic unit-cell parameters of a = 67.0, b = 78.1, c = 92.1 A, alpha = 102.4, beta = 93.5, gamma = 91.7 degrees . A full diffraction data set to 1.33 A resolution has been collected for the wild-type enzyme, as measured from flash-cooled crystals at 100 K, using synchrotron radiation. These data are currently being used for the detailed three-dimensional crystal structure analysis of Gan1D.