Cloning and characterization of extracellular metal protease gene of the algicidal marine bacterium Pseudoalteromonas sp. strain A28.
The gene (empI) encoding an extracellular metal protease was isolated from a Pseudoalteromonas sp. strain A28 DNA library. The recombinant EmpI protein was expressed in E. coli and purified. Paper-disk assays showed that the purified protease had potent algicidal activity. A skim milk-polyacrylamide gel electrophoresis protease assay showed that the 38-kDa band of protease activity, which co-migrated with purified EmpI and was sensitive to 1,10-phenathroline, was detected in the extracellular supernatant of A28.
Lee
Sun-Og
SO
Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University, Higashi-Hiroshima, Japan.
Kato
Junichi
J
Nakashima
Katsunori
K
Kuroda
Akio
A
Ikeda
Tsukasa
T
Takiguchi
Noboru
N
Ohtake
Hisao
H
D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D008670:Metals / Q000378:metabolism*; D008970:Molecular Weight; D031161:Pseudoalteromonas / Q000201:enzymology* / Q000235:genetics*; D015183:Restriction Mapping
D008670:Metals; D010450:Endopeptidases