Purification and characterization of a robust thermostable protease isolated from Bacillus subtilis strain HR02 as an extremozyme
Aim Since the hot water of Genow, a village in Isin rural district in the central district of Bandar Abbas, Hormozgan Province, Iran, has a rich source of thermophilic bacteria, the current study aimed to find a new thermophilic protease enzyme with suitable properties to be used in different indust...
Gespeichert in:
| Veröffentlicht in: | Journal of applied microbiology 2022-11, Vol.133 (5), p.2779-2789 |
|---|---|
| Hauptverfasser: | , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 2789 |
|---|---|
| container_issue | 5 |
| container_start_page | 2779 |
| container_title | Journal of applied microbiology |
| container_volume | 133 |
| creator | Homaei, Ahmad Izadpanah Qeshmi, Fatemeh |
| description | Aim
Since the hot water of Genow, a village in Isin rural district in the central district of Bandar Abbas, Hormozgan Province, Iran, has a rich source of thermophilic bacteria, the current study aimed to find a new thermophilic protease enzyme with suitable properties to be used in different industries.
Methods and results
Water and sediment samples were collected from the hot water of Genow, and finally, 20 colonies were isolated. Among these isolated colonies, two bacterial strains grew on the skim milk agar medium, and a clear halo was formed around the colony, which was accurately identified by 16S rRNA gene sequence analysis. The comparison of 16S rRNA gene sequence analyses of isolated strains HR01 and HR02 with registered sequences of 16S rRNA genes in NCBI showed that the two isolates had the most similarity to Bacillus sonorensis and Bacillus subtilis, respectively. Among the two bacterial strains, the highest enzymatic activity was observed in B. subtilis strain HR02, from which the protease purification process was performed. A putative native B. subtilis strain HR02 protease (BSHR02PR) was purified by the UNO Q‐6 ionic exchange chromatography method. Biochemical analyses revealed a monomeric enzyme, BsHR02Pro, with a molecular weight of 25 kDa, showing the maximum activity at 70°C and pH 8.0. Moreover, the purified enzyme was stable up to 80 °C and in a pH range of 6.0–12.0. The steady‐state kinetic analysis for colloidal casein showed that the Km, Vmax and kcat values of the purified enzyme were 25.7 μM, 93.2 μM min−1 and 2.18 s−1, respectively.
Conclusion
The hot water of Genow is a rich source of protease‐producing bacteria. Sediments are a better source for the isolation of these types of bacteria than spring water. Overall, our results demonstrated a potential bacterial enzyme BsHR02Pro as a suitable catalyst to be used in the various industries. |
| doi_str_mv | 10.1111/jam.15725 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_2693782079</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2726071493</sourcerecordid><originalsourceid>FETCH-LOGICAL-c3305-e7e2bdfbe59587bffa00bd0860b2408e6c5e53b0621849b9cac19c80b86c0ad63</originalsourceid><addsrcrecordid>eNp1kUFrFjEQhhdRsFYP_oOAFz1sO8l-SXaPtVirVFpEz2GSnaX5yG5qkkW_3vrPjV1PgnOZl-GZmRfepnnN4YTXOt3jfMKlFvJJc8Q7JVuhtHj6qHetBC2eNy9y3gPwDqQ6ah5u1uQn77D4uDBcRuZuMaErlPz9NowTQ5aiXXNh5ZbSHHNBG4jdpVgIMzGfY8BCI5tSnNl7dD6ENbO82uKDr6Ik9Au7_AqCYa5fGP0qieZ4f5jpZfNswpDp1d9-3Hy_-PDt_LK9uv746fzsqnVdtdqSJmHHyZIcZK_tNCGAHaFXYMUOelJOkuwsKMH73WAHh44PrgfbKwc4qu64ebvdrbZ_rJSLmX12FAIuFNdshBo63QvQQ0Xf_IPu45qW6s4ILRRovhu6Sr3bKJdizokmc5f8jOlgOJg_YZgahnkMo7KnG_vTBzr8HzSfz75sG78B6YyNXw</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2726071493</pqid></control><display><type>article</type><title>Purification and characterization of a robust thermostable protease isolated from Bacillus subtilis strain HR02 as an extremozyme</title><source>Oxford University Press Journals All Titles (1996-Current)</source><source>Wiley Online Library All Journals</source><creator>Homaei, Ahmad ; Izadpanah Qeshmi, Fatemeh</creator><creatorcontrib>Homaei, Ahmad ; Izadpanah Qeshmi, Fatemeh</creatorcontrib><description>Aim
Since the hot water of Genow, a village in Isin rural district in the central district of Bandar Abbas, Hormozgan Province, Iran, has a rich source of thermophilic bacteria, the current study aimed to find a new thermophilic protease enzyme with suitable properties to be used in different industries.
Methods and results
Water and sediment samples were collected from the hot water of Genow, and finally, 20 colonies were isolated. Among these isolated colonies, two bacterial strains grew on the skim milk agar medium, and a clear halo was formed around the colony, which was accurately identified by 16S rRNA gene sequence analysis. The comparison of 16S rRNA gene sequence analyses of isolated strains HR01 and HR02 with registered sequences of 16S rRNA genes in NCBI showed that the two isolates had the most similarity to Bacillus sonorensis and Bacillus subtilis, respectively. Among the two bacterial strains, the highest enzymatic activity was observed in B. subtilis strain HR02, from which the protease purification process was performed. A putative native B. subtilis strain HR02 protease (BSHR02PR) was purified by the UNO Q‐6 ionic exchange chromatography method. Biochemical analyses revealed a monomeric enzyme, BsHR02Pro, with a molecular weight of 25 kDa, showing the maximum activity at 70°C and pH 8.0. Moreover, the purified enzyme was stable up to 80 °C and in a pH range of 6.0–12.0. The steady‐state kinetic analysis for colloidal casein showed that the Km, Vmax and kcat values of the purified enzyme were 25.7 μM, 93.2 μM min−1 and 2.18 s−1, respectively.
Conclusion
The hot water of Genow is a rich source of protease‐producing bacteria. Sediments are a better source for the isolation of these types of bacteria than spring water. Overall, our results demonstrated a potential bacterial enzyme BsHR02Pro as a suitable catalyst to be used in the various industries.</description><identifier>ISSN: 1364-5072</identifier><identifier>EISSN: 1365-2672</identifier><identifier>DOI: 10.1111/jam.15725</identifier><language>eng</language><publisher>Cambridge: Oxford University Press</publisher><subject>Bacillus subtilis ; Bacteria ; biochemical characterization ; Casein ; Catalysts ; Colonies ; DNA sequencing ; Enzymatic activity ; Enzymes ; Extremozymes ; Hot water ; marine microorganisms ; Molecular weight ; pH effects ; Protease ; protease enzymes ; protein purification ; Proteinase ; Purification ; rRNA 16S ; Sediment samplers ; Sediments ; Sequence analysis ; Skim milk ; Spring water ; Strains (organisms) ; Thermophilic bacteria ; thermostability</subject><ispartof>Journal of applied microbiology, 2022-11, Vol.133 (5), p.2779-2789</ispartof><rights>2022 Society for Applied Microbiology.</rights><rights>Copyright © 2022 The Society for Applied Microbiology</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3305-e7e2bdfbe59587bffa00bd0860b2408e6c5e53b0621849b9cac19c80b86c0ad63</citedby><cites>FETCH-LOGICAL-c3305-e7e2bdfbe59587bffa00bd0860b2408e6c5e53b0621849b9cac19c80b86c0ad63</cites><orcidid>0000-0003-4310-0637</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Fjam.15725$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Fjam.15725$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,780,784,1417,27924,27925,45574,45575</link.rule.ids></links><search><creatorcontrib>Homaei, Ahmad</creatorcontrib><creatorcontrib>Izadpanah Qeshmi, Fatemeh</creatorcontrib><title>Purification and characterization of a robust thermostable protease isolated from Bacillus subtilis strain HR02 as an extremozyme</title><title>Journal of applied microbiology</title><description>Aim
Since the hot water of Genow, a village in Isin rural district in the central district of Bandar Abbas, Hormozgan Province, Iran, has a rich source of thermophilic bacteria, the current study aimed to find a new thermophilic protease enzyme with suitable properties to be used in different industries.
Methods and results
Water and sediment samples were collected from the hot water of Genow, and finally, 20 colonies were isolated. Among these isolated colonies, two bacterial strains grew on the skim milk agar medium, and a clear halo was formed around the colony, which was accurately identified by 16S rRNA gene sequence analysis. The comparison of 16S rRNA gene sequence analyses of isolated strains HR01 and HR02 with registered sequences of 16S rRNA genes in NCBI showed that the two isolates had the most similarity to Bacillus sonorensis and Bacillus subtilis, respectively. Among the two bacterial strains, the highest enzymatic activity was observed in B. subtilis strain HR02, from which the protease purification process was performed. A putative native B. subtilis strain HR02 protease (BSHR02PR) was purified by the UNO Q‐6 ionic exchange chromatography method. Biochemical analyses revealed a monomeric enzyme, BsHR02Pro, with a molecular weight of 25 kDa, showing the maximum activity at 70°C and pH 8.0. Moreover, the purified enzyme was stable up to 80 °C and in a pH range of 6.0–12.0. The steady‐state kinetic analysis for colloidal casein showed that the Km, Vmax and kcat values of the purified enzyme were 25.7 μM, 93.2 μM min−1 and 2.18 s−1, respectively.
Conclusion
The hot water of Genow is a rich source of protease‐producing bacteria. Sediments are a better source for the isolation of these types of bacteria than spring water. Overall, our results demonstrated a potential bacterial enzyme BsHR02Pro as a suitable catalyst to be used in the various industries.</description><subject>Bacillus subtilis</subject><subject>Bacteria</subject><subject>biochemical characterization</subject><subject>Casein</subject><subject>Catalysts</subject><subject>Colonies</subject><subject>DNA sequencing</subject><subject>Enzymatic activity</subject><subject>Enzymes</subject><subject>Extremozymes</subject><subject>Hot water</subject><subject>marine microorganisms</subject><subject>Molecular weight</subject><subject>pH effects</subject><subject>Protease</subject><subject>protease enzymes</subject><subject>protein purification</subject><subject>Proteinase</subject><subject>Purification</subject><subject>rRNA 16S</subject><subject>Sediment samplers</subject><subject>Sediments</subject><subject>Sequence analysis</subject><subject>Skim milk</subject><subject>Spring water</subject><subject>Strains (organisms)</subject><subject>Thermophilic bacteria</subject><subject>thermostability</subject><issn>1364-5072</issn><issn>1365-2672</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2022</creationdate><recordtype>article</recordtype><recordid>eNp1kUFrFjEQhhdRsFYP_oOAFz1sO8l-SXaPtVirVFpEz2GSnaX5yG5qkkW_3vrPjV1PgnOZl-GZmRfepnnN4YTXOt3jfMKlFvJJc8Q7JVuhtHj6qHetBC2eNy9y3gPwDqQ6ah5u1uQn77D4uDBcRuZuMaErlPz9NowTQ5aiXXNh5ZbSHHNBG4jdpVgIMzGfY8BCI5tSnNl7dD6ENbO82uKDr6Ik9Au7_AqCYa5fGP0qieZ4f5jpZfNswpDp1d9-3Hy_-PDt_LK9uv746fzsqnVdtdqSJmHHyZIcZK_tNCGAHaFXYMUOelJOkuwsKMH73WAHh44PrgfbKwc4qu64ebvdrbZ_rJSLmX12FAIuFNdshBo63QvQQ0Xf_IPu45qW6s4ILRRovhu6Sr3bKJdizokmc5f8jOlgOJg_YZgahnkMo7KnG_vTBzr8HzSfz75sG78B6YyNXw</recordid><startdate>202211</startdate><enddate>202211</enddate><creator>Homaei, Ahmad</creator><creator>Izadpanah Qeshmi, Fatemeh</creator><general>Oxford University Press</general><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7QO</scope><scope>7T7</scope><scope>7TM</scope><scope>7U7</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0003-4310-0637</orcidid></search><sort><creationdate>202211</creationdate><title>Purification and characterization of a robust thermostable protease isolated from Bacillus subtilis strain HR02 as an extremozyme</title><author>Homaei, Ahmad ; Izadpanah Qeshmi, Fatemeh</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3305-e7e2bdfbe59587bffa00bd0860b2408e6c5e53b0621849b9cac19c80b86c0ad63</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2022</creationdate><topic>Bacillus subtilis</topic><topic>Bacteria</topic><topic>biochemical characterization</topic><topic>Casein</topic><topic>Catalysts</topic><topic>Colonies</topic><topic>DNA sequencing</topic><topic>Enzymatic activity</topic><topic>Enzymes</topic><topic>Extremozymes</topic><topic>Hot water</topic><topic>marine microorganisms</topic><topic>Molecular weight</topic><topic>pH effects</topic><topic>Protease</topic><topic>protease enzymes</topic><topic>protein purification</topic><topic>Proteinase</topic><topic>Purification</topic><topic>rRNA 16S</topic><topic>Sediment samplers</topic><topic>Sediments</topic><topic>Sequence analysis</topic><topic>Skim milk</topic><topic>Spring water</topic><topic>Strains (organisms)</topic><topic>Thermophilic bacteria</topic><topic>thermostability</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Homaei, Ahmad</creatorcontrib><creatorcontrib>Izadpanah Qeshmi, Fatemeh</creatorcontrib><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Nucleic Acids Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of applied microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Homaei, Ahmad</au><au>Izadpanah Qeshmi, Fatemeh</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Purification and characterization of a robust thermostable protease isolated from Bacillus subtilis strain HR02 as an extremozyme</atitle><jtitle>Journal of applied microbiology</jtitle><date>2022-11</date><risdate>2022</risdate><volume>133</volume><issue>5</issue><spage>2779</spage><epage>2789</epage><pages>2779-2789</pages><issn>1364-5072</issn><eissn>1365-2672</eissn><abstract>Aim
Since the hot water of Genow, a village in Isin rural district in the central district of Bandar Abbas, Hormozgan Province, Iran, has a rich source of thermophilic bacteria, the current study aimed to find a new thermophilic protease enzyme with suitable properties to be used in different industries.
Methods and results
Water and sediment samples were collected from the hot water of Genow, and finally, 20 colonies were isolated. Among these isolated colonies, two bacterial strains grew on the skim milk agar medium, and a clear halo was formed around the colony, which was accurately identified by 16S rRNA gene sequence analysis. The comparison of 16S rRNA gene sequence analyses of isolated strains HR01 and HR02 with registered sequences of 16S rRNA genes in NCBI showed that the two isolates had the most similarity to Bacillus sonorensis and Bacillus subtilis, respectively. Among the two bacterial strains, the highest enzymatic activity was observed in B. subtilis strain HR02, from which the protease purification process was performed. A putative native B. subtilis strain HR02 protease (BSHR02PR) was purified by the UNO Q‐6 ionic exchange chromatography method. Biochemical analyses revealed a monomeric enzyme, BsHR02Pro, with a molecular weight of 25 kDa, showing the maximum activity at 70°C and pH 8.0. Moreover, the purified enzyme was stable up to 80 °C and in a pH range of 6.0–12.0. The steady‐state kinetic analysis for colloidal casein showed that the Km, Vmax and kcat values of the purified enzyme were 25.7 μM, 93.2 μM min−1 and 2.18 s−1, respectively.
Conclusion
The hot water of Genow is a rich source of protease‐producing bacteria. Sediments are a better source for the isolation of these types of bacteria than spring water. Overall, our results demonstrated a potential bacterial enzyme BsHR02Pro as a suitable catalyst to be used in the various industries.</abstract><cop>Cambridge</cop><pub>Oxford University Press</pub><doi>10.1111/jam.15725</doi><tpages>11</tpages><orcidid>https://orcid.org/0000-0003-4310-0637</orcidid></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1364-5072 |
| ispartof | Journal of applied microbiology, 2022-11, Vol.133 (5), p.2779-2789 |
| issn | 1364-5072 1365-2672 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_2693782079 |
| source | Oxford University Press Journals All Titles (1996-Current); Wiley Online Library All Journals |
| subjects | Bacillus subtilis Bacteria biochemical characterization Casein Catalysts Colonies DNA sequencing Enzymatic activity Enzymes Extremozymes Hot water marine microorganisms Molecular weight pH effects Protease protease enzymes protein purification Proteinase Purification rRNA 16S Sediment samplers Sediments Sequence analysis Skim milk Spring water Strains (organisms) Thermophilic bacteria thermostability |
| title | Purification and characterization of a robust thermostable protease isolated from Bacillus subtilis strain HR02 as an extremozyme |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-07T22%3A17%3A22IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Purification%20and%20characterization%20of%20a%20robust%20thermostable%20protease%20isolated%20from%20Bacillus%20subtilis%20strain%20HR02%20as%20an%20extremozyme&rft.jtitle=Journal%20of%20applied%20microbiology&rft.au=Homaei,%20Ahmad&rft.date=2022-11&rft.volume=133&rft.issue=5&rft.spage=2779&rft.epage=2789&rft.pages=2779-2789&rft.issn=1364-5072&rft.eissn=1365-2672&rft_id=info:doi/10.1111/jam.15725&rft_dat=%3Cproquest_cross%3E2726071493%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=2726071493&rft_id=info:pmid/&rfr_iscdi=true |