Open AccessSequencing, cloning and expression of a β‐1,4‐mannanase gene, manA , from the extremely thermophilic anaerobic bacterium, Caldicellulosiruptor Rt8B.4
Author(s) -
Gibbs Moreland D.,
Elinder Anna U.,
Reeves Rosalind A.,
Bergquist Peter L.
Publication year - 1996
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1996.tb08360.x
Subject(s) - thermophile , cloning (programming) , biology , escherichia coli , bacteria , gene , hyperthermophile , microbiology and biotechnology , mesophile , molecular cloning , biochemistry , genetics , archaea , gene expression , computer science , programming language
A gene encoding a β‐mannanase ( manA ) has been cloned from an obligately anaerobic extreme thermophile, Caldicellulosiruptor strain Rt8B.4, which is most closely related to Caldicellulosiruptor saccharolyticus (formerly Caldocellum saccharolyticum ). The gene codes for a multidomain enzyme with a C‐terminal β‐mannanase domain which was amplified by the polymerase chain reaction and cloned into a temperature‐inducible expression vector in Escherichia coli . Sequence comparisons have shown that the Man domain of Rt8B.4 ManA is related to a thermophilic Dictyoglomus mannanase and a mesophilic mannanase from a Bacillus species. It appears to be unrelated to the β‐mannanase domain of C. saccharolyticus , implying acquisition of the genes from unrelated sources by the two bacteria.