Part:BBa_K1175006

endo-beta-1,3-1,4 glucanase (BglS) from Bacillus Subtilis 168

BglS This gene encodes an endo-beta-1,3-1,4-glucanase (bglS), which is from the bacterium Bacillus subtilis subtilis 168. The enzyme will hydrolyze and thereby cleave internal 1,4 linkages adjacent to 1,3 linkages.

Usage and Biology The substrates vulnerable to the bglS encoded enzyme are mixed linked beta-glucans. These glucans would have 1,3 and 1,4 beta linkages within the polysaccharide linking together the glucose monomers. Examples of these glucans can be found in oats, maize, and barley.

(1) http://mic.sgmjournals.org/content/141/2/281.long (2) http://onlinelibrary.wiley.com/doi/10.1002/j.2050-0416.1984.tb04259.x/pdf

bglS

The endo-1,3-1,4-glucanase bglS is a globular protein that that has two residues of interest. The putative nucleophile and acid-base cleavage sites at the E residues 133 and 137 highlighted in red.

bglS Enzyme Activity

Table 2. Substrate specificity of 1,3-1,4-beta-glucanase (BglS) purified to electrophoretic homogenity from E. coli cells harboring recombinant plasmid pRB33. Enzyme activities were calculated from the results of three independent measurements. *Unit defined as 1 micromole reducing sugar min-1 (mg purified enzyme)-1. **Unit defined as 1 OD595 unit min-1 (mg purified enzyme)-1. Table and data from: http://mic.sgmjournals.org/content/141/2/281.long The plates shown are differential medias containing both lichenan ,able to be broken down by the Bacillus Subtilis Subtilis 168 BglS gene. The plates were stained by flooding the plate with Congo red and the visualization of clearing zones was improved by flooding the stained plates with 1M NaCl. The Upper plate indicates, due to an apparent lack of clearing zones, that the strains MW14 (Deleted BglS), MW10 (Deleted EglS102 and Bgl BglSRV) and MW9 (Deletion of EglS and BglS) do not contain a significant ability to break down Lichenan. Whereas the strains MW8 (Deletion of EglS) and DB104 (No deletions) were able to break down the lichenan. The lower plate indicates, due to an apparent lack of clearing zones, that the strains MW10, MW8, and MW9 did not contain a significant ability to break down the CM-cellulose. On the other side of the coin, however, the strains MW14 and DB104 did have the ability to produce clearing zones, indicating their ability to break down CM-cellulose.

bglS on Cellulose Plates

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]