title: Sequencing, cloning and expression of a beta-1,4-mannanase gene, manA, from the extremely thermophilic anaerobic bacterium, Caldicellulosiruptor Rt8B.4.
abstract: A gene encoding a beta-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 beta-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 beta-mannanase domain of C. saccharolyticus, implying acquisition of the genes from unrelated sources by the two bacteria.
authors.affiliation: Centre for Gene Technology, School of Biological Sciences, University of Auckland, New Zealand.
authors.forename: M D
authors.initials: MD
authors.lastname: Gibbs
authors.forename: A U
authors.initials: AU
authors.lastname: Elinder
authors.forename: R A
authors.initials: RA
authors.lastname: Reeves
authors.forename: P L
authors.initials: PL
authors.lastname: Bergquist
mesh_terms: D000595:Amino Acid Sequence; D001421:Bacteria, Anaerobic / Q000201:enzymology / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial / Q000235:genetics*; D008361:Mannosidases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
chemical_list: D011993:Recombinant Fusion Proteins; D008361:Mannosidases; C041867:endo-1,4-beta-D-mannanase