Cloning, expression and decoding of the cold adaptation of a new widely represented thermolabile subtilisin-like protease
Artículo de publicación ISI Aims Cloning, expression and characterization of a new cold-adapted protease with potential biotechnological application, isolated from Antarctic bacteria. Method and Results A subtilisin-like gene was isolated from several Antarctic bacterial genus using CODPEHOP-designe...
Published in: | Journal of Applied Microbiology |
---|---|
Main Author: | |
Format: | Article in Journal/Newspaper |
Language: | English |
Published: |
WILEY-BLACKWELL
2013
|
Subjects: | |
Online Access: | https://doi.org/10.1111/jam.12033 https://repositorio.uchile.cl/handle/2250/125785 |
id |
ftunivchile:oai:repositorio.uchile.cl:2250/125785 |
---|---|
record_format |
openpolar |
spelling |
ftunivchile:oai:repositorio.uchile.cl:2250/125785 2023-05-15T13:41:32+02:00 Cloning, expression and decoding of the cold adaptation of a new widely represented thermolabile subtilisin-like protease Acevedo, J. P. 2013-02 application/pdf https://doi.org/10.1111/jam.12033 https://repositorio.uchile.cl/handle/2250/125785 en eng WILEY-BLACKWELL JOURNAL OF APPLIED MICROBIOLOGY Volume: 114 Issue: 2 Pages: 352-363 Published: FEB 2013 doi:10.1111/jam.12033 https://repositorio.uchile.cl/handle/2250/125785 cold adaption Artículo de revista 2013 ftunivchile https://doi.org/10.1111/jam.12033 2023-03-05T00:50:44Z Artículo de publicación ISI Aims Cloning, expression and characterization of a new cold-adapted protease with potential biotechnological application, isolated from Antarctic bacteria. Method and Results A subtilisin-like gene was isolated from several Antarctic bacterial genus using CODPEHOP-designed primers and a genome walking method. This gene encodes a precursor protein, which undergoes an autocatalytic cleavage resulting in a 34.6 kDa active cold-adapted protease with a maximum activity at 2535 degrees C and optimum pH of 8.09.0. It showed a higher catalytic efficiency at lower temperatures compared to its mesophilic counterpart. Heat-induced inactivation resulted in a very low melting point. Local packing analysis using the homology model indicated Ala284 as an important cold-adaptation determinant, which was corroborated by the site-directed mutagenesis. Conclusions A new thermolabile subtilisin-like protease has been successfully cloned and analysed, and an important hot spot in the evolution of the cold adaptation and substrate specificity of this enzyme was identified and tested. Significance and Impact of the Study This work reports a new cold-adapted protease with a vast representation amongst Antarctic genus, suggesting therefore its evolutionary success in this cold environment. Likewise, important sites for genetic potentiation have been identified, which are extrapolated to other enzymes of the same kind. FONDEF DO4I-1374 Conicyt Millenium Scientific Initiative (ICDB) P05-001-F Article in Journal/Newspaper Antarc* Antarctic Universidad de Chile: Repositorio académico Antarctic Isi ENVELOPE(-38.550,-38.550,65.617,65.617) Journal of Applied Microbiology 114 2 352 363 |
institution |
Open Polar |
collection |
Universidad de Chile: Repositorio académico |
op_collection_id |
ftunivchile |
language |
English |
topic |
cold adaption |
spellingShingle |
cold adaption Acevedo, J. P. Cloning, expression and decoding of the cold adaptation of a new widely represented thermolabile subtilisin-like protease |
topic_facet |
cold adaption |
description |
Artículo de publicación ISI Aims Cloning, expression and characterization of a new cold-adapted protease with potential biotechnological application, isolated from Antarctic bacteria. Method and Results A subtilisin-like gene was isolated from several Antarctic bacterial genus using CODPEHOP-designed primers and a genome walking method. This gene encodes a precursor protein, which undergoes an autocatalytic cleavage resulting in a 34.6 kDa active cold-adapted protease with a maximum activity at 2535 degrees C and optimum pH of 8.09.0. It showed a higher catalytic efficiency at lower temperatures compared to its mesophilic counterpart. Heat-induced inactivation resulted in a very low melting point. Local packing analysis using the homology model indicated Ala284 as an important cold-adaptation determinant, which was corroborated by the site-directed mutagenesis. Conclusions A new thermolabile subtilisin-like protease has been successfully cloned and analysed, and an important hot spot in the evolution of the cold adaptation and substrate specificity of this enzyme was identified and tested. Significance and Impact of the Study This work reports a new cold-adapted protease with a vast representation amongst Antarctic genus, suggesting therefore its evolutionary success in this cold environment. Likewise, important sites for genetic potentiation have been identified, which are extrapolated to other enzymes of the same kind. FONDEF DO4I-1374 Conicyt Millenium Scientific Initiative (ICDB) P05-001-F |
format |
Article in Journal/Newspaper |
author |
Acevedo, J. P. |
author_facet |
Acevedo, J. P. |
author_sort |
Acevedo, J. P. |
title |
Cloning, expression and decoding of the cold adaptation of a new widely represented thermolabile subtilisin-like protease |
title_short |
Cloning, expression and decoding of the cold adaptation of a new widely represented thermolabile subtilisin-like protease |
title_full |
Cloning, expression and decoding of the cold adaptation of a new widely represented thermolabile subtilisin-like protease |
title_fullStr |
Cloning, expression and decoding of the cold adaptation of a new widely represented thermolabile subtilisin-like protease |
title_full_unstemmed |
Cloning, expression and decoding of the cold adaptation of a new widely represented thermolabile subtilisin-like protease |
title_sort |
cloning, expression and decoding of the cold adaptation of a new widely represented thermolabile subtilisin-like protease |
publisher |
WILEY-BLACKWELL |
publishDate |
2013 |
url |
https://doi.org/10.1111/jam.12033 https://repositorio.uchile.cl/handle/2250/125785 |
long_lat |
ENVELOPE(-38.550,-38.550,65.617,65.617) |
geographic |
Antarctic Isi |
geographic_facet |
Antarctic Isi |
genre |
Antarc* Antarctic |
genre_facet |
Antarc* Antarctic |
op_relation |
JOURNAL OF APPLIED MICROBIOLOGY Volume: 114 Issue: 2 Pages: 352-363 Published: FEB 2013 doi:10.1111/jam.12033 https://repositorio.uchile.cl/handle/2250/125785 |
op_doi |
https://doi.org/10.1111/jam.12033 |
container_title |
Journal of Applied Microbiology |
container_volume |
114 |
container_issue |
2 |
container_start_page |
352 |
op_container_end_page |
363 |
_version_ |
1766151772410216448 |