The DNA sequence of the gene and genetic control sites for the excretedB. subtilis enzymeβglucanase

Abstract

The sequence of a 1409 base pair restriction fragment containing the B. subtilis gene for (1-3), (1-4)-beta-D-glucan endoglucanase is reported. The gene is encoded in a 726 base pair segment. The deduced amino acid sequence of the protein has a hydrophobic signal peptide at the NH2-terminus similar to those found in five other secreted proteins from Bacillus. The gene is preceded by a sequence resembling promoters for the vegetative B. subtilis RNA polymerase. This is followed by a sequence resembling a B. subtilis ribosome binding site nine nucleotides before the first codon of the gene. Two sequences, one before and one after the gene, can be arranged in secondary structures similar to transcriptional terminators. There is also a short open reading frame coding for a hydrophobic protein on the minus strand just upstream from the beta-glucanase gene. A possible role for this gene in the control of expression of beta-glucanase is suggested.

Crystal structure of Bacillus licheniformis 1,3‐1,4‐β‐d‐glucan 4‐glucanohydrolase at 1.8 Å resolution

“…3.2.1.73) from different Bacillus strains have been cloned and sequenced [1][2][3][4][5][6][7]. These enzymes cleave fl-1,4-glycosidic bonds that are adjacent tofl-l,3-glycosidic bonds in mixed linked glucans.…”

Molecular and active-site structure of a Bacillus 1,3-1,4-beta-glucanase.

“…1A). The ,B-glucanases secreted from different Bacillus strains share significant sequence homology (10)(11)(12)(13)(14) and also with the enzymes from Clostridium thermocellum and from Fibrobacter succinogenes (15,16).…”

Construction of a yeast vector directing the synthesis and release of barley (1→3, 1→4)-β-glucanase

“…The chimaeric cDNA gene was introduced in Saccharomyces cerevisiae via the expression vector pMA56 and it was shown that the barley gene is expressed and the gene product exported from the cells to the culture medium. The only other (1---~3, l---~4)-13-glucanase for which the primary structure is known is the Bacillus subtilis enzyme (28). This gene has also been cloned in yeast -E. coil shuttle vectors and introduced into yeast by two groups (9,18,19).…”