@article{oai:nipr.repo.nii.ac.jp:00010175,
 author = {D., de Pascale and M., Giuliani and C., De Santi and N., Bergamasco and A., Amoresano and A., Carpentieri and E., Parrilli and M.L., Tutino and D., de Pascale and M., Giuliani and C., De Santi and N., Bergamasco and A., Amoresano and A., Carpentieri and E., Parrilli and M.L., Tutino},
 issue = {2},
 journal = {Polar science, Polar science},
 month = {Aug},
 note = {Cold-adapted proteases have been found to be the dominant activity throughout the cold marine environment, indicating their importance in bacterial acquisition of nitrogen-rich complex organic compounds. However, few extracellular proteases from marine organisms have been characterized so far, and the mechanisms that enable their activity in situ are still largely unknown. Aside from their ecological importance and use as model enzyme for structure/function investigations, cold-active proteolytic enzymes offer great potential for biotechnological applications.Our studies on cold adapted proteases were performed on exo-enzyme produced by the Antarctic marine bacterium Pseudoalteromonas haloplanktis TAC125. By applying a proteomic approach, we identified several proteolytic activities from its culture supernatant. PhAP protease was selected for further investigations. The encoding gene was cloned and the protein was recombinantly produced in E. coli cells. The homogeneous product was biochemically characterised and it turned out that the enzyme is a Zn-dependent aminopeptidase, with an activity dependence from assay temperature typical of psychrophilic enzymes., 南極の海洋性バクテリアであるPseudoalteromonas haloplanktis TAC125により生産される細胞外酵素について、タンパク質分解酵素の低温適応を調べた。プロテオミクス解析により、その培養液の上澄みからいくつかのタンパク分解活性を同定した。以後の解析にはPhAPプロテアーゼを選び、その遺伝子をクローニングして遺伝子組み換え技術を用い、大腸菌細胞中で生産させた。得られたプロテアーゼの生化学的特徴を調べたところ、酵素は亜鉛依存型のアミノペプチドであり、その活性は典型的な好冷性酵素の特徴を示すことが明らかになった。},
 pages = {285--294},
 title = {PhAP protease from Pseudoalteromonas haloplanktis TAC125: Gene cloning, recombinant production in E. coli and enzyme characterization},
 volume = {4},
 year = {2010}
}