Cloning of two genes from Bacillus circulans WL-12 which encode 1,3-beta-glucanase activity.
Two genes encoding distinct 1,3-beta-glucanases have been cloned from Bacillus circulans and expressed in Escherichia coli. A cosmid library of B. circulans WL-12 DNA was constructed in the broad-host-range cosmid pLAFR1 and screened in E. coli for clones which exhibited 1,3-beta-glucanase activity. Two 1,3-beta-glucanase-positive clones were identified which contained genes encoding two independent 1,3-beta-glucanases as shown by biochemical, physical and molecular analyses. The cosmids, designated pMON5401 (27.1 kb insert) and pMON5402 (24.7 kb insert), encoded 68 kDa and 40 kDa 1,3-beta-glucanases, respectively. Both 1,3-beta-glucanases were purified from their respective E. coli strains, characterized biochemically, and were shown to exhibit lytic activity against purified yeast cell wall preparations.
Fiske
M J
MJ
Monsanto Company, St. Louis, Missouri 63198.
Tobey-Fincher
K L
KL
Fuchs
R L
RL
D000595:Amino Acid Sequence; D001407:Bacillus / Q000201:enzymology / Q000235:genetics*; D003001:Cloning, Molecular; D003360:Cosmids; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005798:Genes, Bacterial; D043326:Glucan 1,3-beta-Glucosidase; D008969:Molecular Sequence Data; D008970:Molecular Weight; D001617:beta-Glucosidase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism
D001617:beta-Glucosidase; D043326:Glucan 1,3-beta-Glucosidase