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A novel thiol-dependent serine protease from Neocosmospora sp. N1

Alkaline proteases have several industrial applications. In the present study, newly isolated Neocosmospora sp. N1 was screened as hyper producer of serine protease. A multimeric protease of the fungus was purified to homogeneity till 96.78 fold purification with 22.51% recovery. The homogeneity of...

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Autores principales: Matkawala, Fatema, Nighojkar, Sadhana, Kumar, Anil, Nighojkar, Anand
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6699422/
https://www.ncbi.nlm.nih.gov/pubmed/31440596
http://dx.doi.org/10.1016/j.heliyon.2019.e02246
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author Matkawala, Fatema
Nighojkar, Sadhana
Kumar, Anil
Nighojkar, Anand
author_facet Matkawala, Fatema
Nighojkar, Sadhana
Kumar, Anil
Nighojkar, Anand
author_sort Matkawala, Fatema
collection PubMed
description Alkaline proteases have several industrial applications. In the present study, newly isolated Neocosmospora sp. N1 was screened as hyper producer of serine protease. A multimeric protease of the fungus was purified to homogeneity till 96.78 fold purification with 22.51% recovery. The homogeneity of purified enzyme was checked by native PAGE and its molecular weight was found to be 198.03 kDa by MALDI-TOF. On SDS-PAGE analysis, enzyme was found to be a hetero oligomer of 17.66 kDa and 20.89 kDa subunits. The purified enzyme showed maximum activity with casein as substrate at 60 °C and pH 8.5. The K(m) and V(max) values were found to be 0.015 mg/ml and 454.45 U/ml, respectively. The enzyme was completely inhibited by PMSF, while the activity was 40% enhanced using β-mercaptoethanol, suggesting that it is a thiol-dependent serine protease. The purified protease was active over an alkaline pH range from 7 to 12 and temperatures from 20 °C to 60 °C. The enzyme exhibited excellent stability, almost 100% towards organic solvents such as toluene, benzene and hexane, surfactants such as Triton X-100, Tween-20, Tween-80 and SDS, as well as commercial detergents. The significant properties of purified enzyme assure that it could be a potential candidate for commercial purposes.
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spelling pubmed-66994222019-08-22 A novel thiol-dependent serine protease from Neocosmospora sp. N1 Matkawala, Fatema Nighojkar, Sadhana Kumar, Anil Nighojkar, Anand Heliyon Article Alkaline proteases have several industrial applications. In the present study, newly isolated Neocosmospora sp. N1 was screened as hyper producer of serine protease. A multimeric protease of the fungus was purified to homogeneity till 96.78 fold purification with 22.51% recovery. The homogeneity of purified enzyme was checked by native PAGE and its molecular weight was found to be 198.03 kDa by MALDI-TOF. On SDS-PAGE analysis, enzyme was found to be a hetero oligomer of 17.66 kDa and 20.89 kDa subunits. The purified enzyme showed maximum activity with casein as substrate at 60 °C and pH 8.5. The K(m) and V(max) values were found to be 0.015 mg/ml and 454.45 U/ml, respectively. The enzyme was completely inhibited by PMSF, while the activity was 40% enhanced using β-mercaptoethanol, suggesting that it is a thiol-dependent serine protease. The purified protease was active over an alkaline pH range from 7 to 12 and temperatures from 20 °C to 60 °C. The enzyme exhibited excellent stability, almost 100% towards organic solvents such as toluene, benzene and hexane, surfactants such as Triton X-100, Tween-20, Tween-80 and SDS, as well as commercial detergents. The significant properties of purified enzyme assure that it could be a potential candidate for commercial purposes. Elsevier 2019-08-09 /pmc/articles/PMC6699422/ /pubmed/31440596 http://dx.doi.org/10.1016/j.heliyon.2019.e02246 Text en © 2019 Published by Elsevier Ltd. http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Article
Matkawala, Fatema
Nighojkar, Sadhana
Kumar, Anil
Nighojkar, Anand
A novel thiol-dependent serine protease from Neocosmospora sp. N1
title A novel thiol-dependent serine protease from Neocosmospora sp. N1
title_full A novel thiol-dependent serine protease from Neocosmospora sp. N1
title_fullStr A novel thiol-dependent serine protease from Neocosmospora sp. N1
title_full_unstemmed A novel thiol-dependent serine protease from Neocosmospora sp. N1
title_short A novel thiol-dependent serine protease from Neocosmospora sp. N1
title_sort novel thiol-dependent serine protease from neocosmospora sp. n1
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6699422/
https://www.ncbi.nlm.nih.gov/pubmed/31440596
http://dx.doi.org/10.1016/j.heliyon.2019.e02246
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