Sequence of egV and properties of EgV, a Ruminococcus albus endoglucanase containing a dockerin domain.
The Ruminococcus albus F-40 egV gene, encoding endoglucanase V (EGV), consists of an open reading frame of 1,833 nucleotides and encodes 611 amino acids with a deduced molecular weight of 67,103. The deduced EGV is a modular enzyme composed of a catalytic domain of family 5 of glycosyl hydrolases, a domain of unknown function, and a dockerin domain responsible for cellulosome assembly, suggesting that R. albus F-40 produces a cellulosome, and EGV is a component of the cellulosome. A truncated form of EGV with an apparent molecular weight of 42,000 was purified from a recombinant Escherichia coli and characterized since EGV suffered from partial proteolysis by E. coli protease(s). The truncated EGV was active toward carboxylmethyl cellulose, xylan, lichenan, and acid-swollen cellulose. The pH and temperature optima of the enzyme were 7.0 and 40 degrees C, respectively. By Western blot analysis using the antiserum raised against the truncated enzyme, EGV was detected in the whole cells but not in the culture supernatant of R. alubus F-40, suggesting that EGV was located on the cell surface.
Ohara
H
H
Faculty of Bioresources, Mie University, Tsu, Japan.
Noguchi
J
J
Karita
S
S
Kimura
T
T
Sakka
K
K
Ohmiya
K
K
D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D001483:Base Sequence; D002480:Cellulase / Q000235:genetics / Q000378:metabolism*; D002855:Chromatography, Thin Layer; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D015723:Gene Library; D006095:Gram-Positive Cocci / Q000201:enzymology* / Q000235:genetics; D008565:Membrane Proteins / Q000737:chemistry; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry
D001426:Bacterial Proteins; D008565:Membrane Proteins; D011994:Recombinant Proteins; C105449:SdbA protein, bacteria; C097231:endoglucanase V; D002480:Cellulase