Identification, recombinant production and partial biochemical characterization of an extracellular cold-active serine-metalloprotease from an Antarctic Pseudomonas isolate
Cold-adapted enzymes are generally derived from psychrophilic microorganisms and have features that make them very attractive for industrial and biotechnological purposes. In this work, we identified a 50 kDa extracellular protease (MP10) from the Antarctic isolate Pseudomonas sp. AU10. The enzyme w...
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| Format: | article |
| Language: | English |
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2017
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| Online Access: | https://hdl.handle.net/20.500.12008/22016 |
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| _version_ | 1868890242386231296 |
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| author | Fullana, Natalia |
| author2 | Braña, Victoria Marizcurrena, Juan José Morales, Danilo Betton, Jean-Michel Marín Gutiérrez, Mónica Castro-Sowinski, Susana |
| author2_role | author author author author author author |
| author_browse | Betton, Jean-Michel Braña, Victoria Castro-Sowinski, Susana Fullana, Natalia Marizcurrena, Juan José Marín Gutiérrez, Mónica Morales, Danilo |
| author_facet | Fullana, Natalia Braña, Victoria Marizcurrena, Juan José Morales, Danilo Betton, Jean-Michel Marín Gutiérrez, Mónica Castro-Sowinski, Susana |
| author_role | author |
| collection | COLIBRI |
| dc.contributor.none.fl_str_mv | Fullana , Natalia. Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología Braña, Victoria. IIBCE Marizcurrena, Juan José. Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología Morales, Danilo. Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología Marín Gutierrez, Mónica. Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología Castro-Sowinski, Susana. Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología |
| dc.creator.none.fl_str_mv | Fullana, Natalia Braña, Victoria Marizcurrena, Juan José Morales, Danilo Betton, Jean-Michel Marín Gutiérrez, Mónica Castro-Sowinski, Susana |
| dc.date.none.fl_str_mv | 2017 2019-10-02T22:08:28Z 2019-10-02T22:08:28Z 20190930 |
| dc.format.none.fl_str_mv | application/pdf |
| dc.identifier.none.fl_str_mv | Fullana, N., et al.Identification, recombinant production and partial biochemical characterization of an extracellular cold-active serine-metalloprotease from an Antarctic Pseudomonas isolate. AIMS Bioengineering, 2017, 4 (3): 386-401. doi: 10.3934/bioeng.2017.3.386 2375-1495 https://hdl.handle.net/20.500.12008/22016 10.3934/bioeng.2017.3.386 |
| dc.language.none.fl_str_mv | en eng |
| dc.publisher.none.fl_str_mv | AIMS Press |
| dc.relation.none.fl_str_mv | AIMS Bioengineering, 2017, 4 (3): 386-401 |
| dc.rights.none.fl_str_mv | info:eu-repo/semantics/openAccess Licencia Creative Commons Atribución (CC –BY 4.0) |
| dc.source.none.fl_str_mv | reponame:COLIBRI instname:Universidad de la República instacron:Universidad de la República |
| dc.subject.none.fl_str_mv | Protease Pseudomonas Cold-adaptation Surfactant Antarctica |
| dc.title.none.fl_str_mv | Identification, recombinant production and partial biochemical characterization of an extracellular cold-active serine-metalloprotease from an Antarctic Pseudomonas isolate |
| dc.type.none.fl_str_mv | Artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion |
| description | Cold-adapted enzymes are generally derived from psychrophilic microorganisms and have features that make them very attractive for industrial and biotechnological purposes. In this work, we identified a 50 kDa extracellular protease (MP10) from the Antarctic isolate Pseudomonas sp. AU10. The enzyme was produced by recombinant DNA technology, purified using immobilized metal affinity chromatography and partially characterized. MP10 is an alkaline thermosensitive serine-metallo protease with optimal activity at pH 8.0 and 40 ℃, in the presence of 1.5 mM Ca2+. MP10 showed 100% residual activity and stability (up to 60 min) when incubated with 7% of non-ionic surfactants (Triton X-100, Tween-80 and Tween-20) and 1.5% of the oxidizing agent hydrogen peroxide. The 3D MP10 structure was predicted and compared with the crystal structure of mesophilic homologous protease produced by Pseudomonas aeruginosa PA01 (reference strain) and other proteases, showing similarity in surface area and volume of proteins, but a significantly higher surface pocket area and volume of MP10. The observed differences presumably may explain the enhanced activity of MP10 for substrate binding at low temperatures. These results give insight to the potential use of MP10 in developing new biotechnologically processes active at low to moderate temperatures, probably with focus in the detergent industry. |
| eu_rights_str_mv | openAccess |
| format | article |
| id | anni_f9fd757efde99cae75a0ca2a98b2b76f |
| identifier_str_mv | Fullana, N., et al.Identification, recombinant production and partial biochemical characterization of an extracellular cold-active serine-metalloprotease from an Antarctic Pseudomonas isolate. AIMS Bioengineering, 2017, 4 (3): 386-401. doi: 10.3934/bioeng.2017.3.386 2375-1495 10.3934/bioeng.2017.3.386 |
| instacron_str | Universidad de la República |
| institution | Universidad de la República |
| instname_str | Universidad de la República |
| language | eng |
| language_invalid_str_mv | en |
| network_acronym_str | anni |
| network_name_str | oai-lr-anni |
| oai_identifier_str | oai:colibri.udelar.edu.uy:20.500.12008/22016 |
| publishDate | 2017 |
| publishDateSort | 2017 |
| publisher.none.fl_str_mv | AIMS Press |
| reponame_str | COLIBRI |
| repository.mail.fl_str_mv | |
| repository.name.fl_str_mv | |
| repository_id_str | |
| rights_invalid_str_mv | Licencia Creative Commons Atribución (CC –BY 4.0) |
| spelling | Identification, recombinant production and partial biochemical characterization of an extracellular cold-active serine-metalloprotease from an Antarctic Pseudomonas isolateFullana, NataliaBraña, VictoriaMarizcurrena, Juan JoséMorales, DaniloBetton, Jean-MichelMarín Gutiérrez, MónicaCastro-Sowinski, SusanaProteasePseudomonasCold-adaptationSurfactantAntarcticaCold-adapted enzymes are generally derived from psychrophilic microorganisms and have features that make them very attractive for industrial and biotechnological purposes. In this work, we identified a 50 kDa extracellular protease (MP10) from the Antarctic isolate Pseudomonas sp. AU10. The enzyme was produced by recombinant DNA technology, purified using immobilized metal affinity chromatography and partially characterized. MP10 is an alkaline thermosensitive serine-metallo protease with optimal activity at pH 8.0 and 40 ℃, in the presence of 1.5 mM Ca2+. MP10 showed 100% residual activity and stability (up to 60 min) when incubated with 7% of non-ionic surfactants (Triton X-100, Tween-80 and Tween-20) and 1.5% of the oxidizing agent hydrogen peroxide. The 3D MP10 structure was predicted and compared with the crystal structure of mesophilic homologous protease produced by Pseudomonas aeruginosa PA01 (reference strain) and other proteases, showing similarity in surface area and volume of proteins, but a significantly higher surface pocket area and volume of MP10. The observed differences presumably may explain the enhanced activity of MP10 for substrate binding at low temperatures. These results give insight to the potential use of MP10 in developing new biotechnologically processes active at low to moderate temperatures, probably with focus in the detergent industry.AIMS PressFullana , Natalia. Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de BiologíaBraña, Victoria. IIBCEMarizcurrena, Juan José. Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de BiologíaMorales, Danilo. Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de BiologíaMarín Gutierrez, Mónica. Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de BiologíaCastro-Sowinski, Susana. Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología2019-10-02T22:08:28Z2019-10-02T22:08:28Z201720190930Artículoinfo:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfFullana, N., et al.Identification, recombinant production and partial biochemical characterization of an extracellular cold-active serine-metalloprotease from an Antarctic Pseudomonas isolate. AIMS Bioengineering, 2017, 4 (3): 386-401. doi: 10.3934/bioeng.2017.3.3862375-1495https://hdl.handle.net/20.500.12008/2201610.3934/bioeng.2017.3.386reponame:COLIBRIinstname:Universidad de la Repúblicainstacron:Universidad de la RepúblicaenengAIMS Bioengineering, 2017, 4 (3): 386-401Las obras depositadas en el Repositorio se rigen por la Ordenanza de los Derechos de la Propiedad Intelectual de la Universidad De La República. (Res. Nº 91 de C.D.C. de 8/III/1994 – D.O. 7/IV/1994) y por la Ordenanza del Repositorio Abierto de la Universidad de la República (Res. Nº 16 de C.D.C. de 07/10/2014)info:eu-repo/semantics/openAccessLicencia Creative Commons Atribución (CC –BY 4.0)oai:colibri.udelar.edu.uy:20.500.12008/220162026-04-14T10:09:15Z |
| spellingShingle | Identification, recombinant production and partial biochemical characterization of an extracellular cold-active serine-metalloprotease from an Antarctic Pseudomonas isolate Fullana, Natalia Protease Pseudomonas Cold-adaptation Surfactant Antarctica |
| status_str | publishedVersion |
| title | Identification, recombinant production and partial biochemical characterization of an extracellular cold-active serine-metalloprotease from an Antarctic Pseudomonas isolate |
| title_full | Identification, recombinant production and partial biochemical characterization of an extracellular cold-active serine-metalloprotease from an Antarctic Pseudomonas isolate |
| title_fullStr | Identification, recombinant production and partial biochemical characterization of an extracellular cold-active serine-metalloprotease from an Antarctic Pseudomonas isolate |
| title_full_unstemmed | Identification, recombinant production and partial biochemical characterization of an extracellular cold-active serine-metalloprotease from an Antarctic Pseudomonas isolate |
| title_short | Identification, recombinant production and partial biochemical characterization of an extracellular cold-active serine-metalloprotease from an Antarctic Pseudomonas isolate |
| title_sort | Identification, recombinant production and partial biochemical characterization of an extracellular cold-active serine-metalloprotease from an Antarctic Pseudomonas isolate |
| topic | Protease Pseudomonas Cold-adaptation Surfactant Antarctica |
| url | https://hdl.handle.net/20.500.12008/22016 |