Cloning and relational analysis of 15 novel fungal endoglucanases from family 12 glycosyl hydrolase.
Cellulases belong to the large family of glycosyl hydrolases (GHs) and are produced by a variety of bacteria and fungi. These extracellular enzymes act as endoglucanases (EGs), cellobiohydrolases or beta-glucosidases. In this paper, we describe molecular screening for EGs from the GH family 12. Using three homologous sequence boxes deduced from five previously known members of the family, we analysed 22 cellulase-producing fungal strains obtained from a diverse area of the fungal kingdom. Polymerase chain reactions using degenerate primers designed to the homologous protein boxes were used to identify the family 12 homologues. Several fungi showed the presence of multiple versions of the gene, while amino acid sequence analysis showed diversity in 15 novel members of the family, ranging from 26% to 96% similarity. Our sequence analysis shows that the phylogenetic tree of family 12 EGs can be divided into four subfamilies: 12-1 (fungal group I), 12-2 (fungal group II), 12-3 ( Streptomyces group in which Rhodothermus marinus fits) and 12-4 ( Thermophiles group). Erwinia carotovora may form a new subgroup.
Goedegebuur
Frits
F
Genencor International B.V., Archimedesweg 30, 2333CN Leiden, The Netherlands. fgoedegebuur@genencor.com
Fowler
Timothy
T
Phillips
Jay
J
van der Kley
Pim
P
van Solingen
Piet
P
Dankmeyer
Lydia
L
Power
Scott D
SD
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D002480:Cellulase / Q000737:chemistry* / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D016386:Chromosome Walking; D003001:Cloning, Molecular; D016384:Consensus Sequence; D004271:DNA, Fungal / Q000235:genetics; D005658:Fungi / Q000145:classification / Q000201:enzymology* / Q000235:genetics*; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D004271:DNA, Fungal; D002480:Cellulase