Unique substrate specificity of a thermostable glycosyl hydrolase from an uncultured Anaerolinea, derived from bacterial mat on a subsurface geothermal water stream.
To investigate novel extremozymes encoded by sequenced metagenes from a microbial community in an extreme environment, we have characterized a recombinant glycosyl hydrolase (rGH) from an uncultured bacterium within the order Chloroflexi. rGH formed insoluble bodies in an Escherichia coli protein expression system. The protein was partially dissolved by a surfactant and was enzymatically characterized. The MW of the monomeric peptide was ~62 kDa, and it formed a homodimers in buffer. It was optimally active at 65 °C and from pH 4 to 8. rGH showed hydrolytic activity for α-1,1, α-1,2 and α-1,6 linkages, including isomaltose, but not α-1,4 and β-linkages.
Institute of Biogeoscience, Japan Agency for Marine-Earth Science and Technology, 2-15 Natsushima-cho, Yokosuka 237-0061, Japan. konishi-masaaki@jamstec.go.jp
Masaaki
M
Konishi
Shinro
S
Nishi
Hideto
H
Takami
Yasuhiro
Y
Shimane
Yuriko
Y
Nagano
Kozue
K
Mori
Yukari
Y
Ohta
Yuji
Y
Hatada
D000595:Amino Acid Sequence; D018441:Biofilms; D041862:Chloroflexi / Q000201:enzymology / Q000502:physiology*; D016000:Cluster Analysis; D004795:Enzyme Stability; D006026:Glycoside Hydrolases / Q000737:chemistry* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D060086:Hydrothermal Vents / Q000382:microbiology*; D007534:Isomaltose / Q000737:chemistry / Q000378:metabolism*; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D020539:Sequence Analysis, Protein; D013379:Substrate Specificity; D013696:Temperature
D007534:Isomaltose; D006026:Glycoside Hydrolases