Structural and functional analyses of an L-asparaginase from Geobacillus thermopakistaniensis
Genome sequence of Geobacillus thermopakistaniensis contains an open reading frame annotated as a type II L-asparaginase (ASNaseGt). Critical structural analysis disclosed that ASNaseGt might be a type I L-asparaginase. In order to determine whether it is a type I or type II L-asparaginase, we have...
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| Veröffentlicht in: | International journal of biological macromolecules 2024-04, Vol.263 (Pt 2), p.130438-130438, Article 130438 |
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| container_issue | Pt 2 |
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| container_title | International journal of biological macromolecules |
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| creator | Sania, Ayesha Muhammad, Majida Atta Sajed, Muhammad Azim, Naseema Ahmad, Nasir Aslam, Mehwish Tang, Xiao-Feng Rashid, Naeem |
| description | Genome sequence of Geobacillus thermopakistaniensis contains an open reading frame annotated as a type II L-asparaginase (ASNaseGt). Critical structural analysis disclosed that ASNaseGt might be a type I L-asparaginase. In order to determine whether it is a type I or type II L-asparaginase, we have performed the structural-functional characterization of the recombinant protein as well as analyzed the localization of ASNaseGt in G. thermopakistaniensis. ASNaseGt exhibited optimal activity at 52 °C and pH 9.5. There was a > 3-fold increase in activity in the presence of β-mercaptoethanol. Apparent Vmax and Km values were 2735 U/mg and 0.35 mM, respectively. ASNaseGt displayed high thermostability with >80 % residual activity even after 6 h of incubation at 55 °C. Recombinant ASNaseGt existed in oligomeric form. Addition of β-mercaptoethanol lowered the degree of oligomerization and displayed that tetrameric form was the most active, with a specific activity of 4300 U/mg. Under physiological conditions, ASNaseGt displayed >50 % of the optimal activity. Localization studies in G. thermopakistaniensis revealed that ASNaseGt is a cytosolic protein. Structural and functional characterization, and localization in G. thermopakistaniensis displayed that ASNaseGt is not a type II but a type I L-asparaginase. |
| doi_str_mv | 10.1016/j.ijbiomac.2024.130438 |
| format | Article |
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Critical structural analysis disclosed that ASNaseGt might be a type I L-asparaginase. In order to determine whether it is a type I or type II L-asparaginase, we have performed the structural-functional characterization of the recombinant protein as well as analyzed the localization of ASNaseGt in G. thermopakistaniensis. ASNaseGt exhibited optimal activity at 52 °C and pH 9.5. There was a > 3-fold increase in activity in the presence of β-mercaptoethanol. Apparent Vmax and Km values were 2735 U/mg and 0.35 mM, respectively. ASNaseGt displayed high thermostability with >80 % residual activity even after 6 h of incubation at 55 °C. Recombinant ASNaseGt existed in oligomeric form. Addition of β-mercaptoethanol lowered the degree of oligomerization and displayed that tetrameric form was the most active, with a specific activity of 4300 U/mg. Under physiological conditions, ASNaseGt displayed >50 % of the optimal activity. Localization studies in G. thermopakistaniensis revealed that ASNaseGt is a cytosolic protein. Structural and functional characterization, and localization in G. thermopakistaniensis displayed that ASNaseGt is not a type II but a type I L-asparaginase.</description><identifier>ISSN: 0141-8130</identifier><identifier>EISSN: 1879-0003</identifier><identifier>DOI: 10.1016/j.ijbiomac.2024.130438</identifier><identifier>PMID: 38408579</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>asparaginase ; Asparaginase - chemistry ; Enzyme Stability ; G. thermopakistaniensis ; Geobacillus ; Geobacillus - genetics ; Geobacillus - metabolism ; L-Asparaginase ; Mercaptoethanol ; nucleotide sequences ; Oligomerization ; recombinant proteins ; Recombinant Proteins - genetics ; Structural analysis ; thermal stability ; Thermostability</subject><ispartof>International journal of biological macromolecules, 2024-04, Vol.263 (Pt 2), p.130438-130438, Article 130438</ispartof><rights>2024 Elsevier B.V.</rights><rights>Copyright © 2024 Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><cites>FETCH-LOGICAL-c348t-6daa95b9c4692b5423605ef0f1f6c28d2648148db5bf408403bf879caeea14013</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0141813024012418$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/38408579$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Sania, Ayesha</creatorcontrib><creatorcontrib>Muhammad, Majida Atta</creatorcontrib><creatorcontrib>Sajed, Muhammad</creatorcontrib><creatorcontrib>Azim, Naseema</creatorcontrib><creatorcontrib>Ahmad, Nasir</creatorcontrib><creatorcontrib>Aslam, Mehwish</creatorcontrib><creatorcontrib>Tang, Xiao-Feng</creatorcontrib><creatorcontrib>Rashid, Naeem</creatorcontrib><title>Structural and functional analyses of an L-asparaginase from Geobacillus thermopakistaniensis</title><title>International journal of biological macromolecules</title><addtitle>Int J Biol Macromol</addtitle><description>Genome sequence of Geobacillus thermopakistaniensis contains an open reading frame annotated as a type II L-asparaginase (ASNaseGt). Critical structural analysis disclosed that ASNaseGt might be a type I L-asparaginase. In order to determine whether it is a type I or type II L-asparaginase, we have performed the structural-functional characterization of the recombinant protein as well as analyzed the localization of ASNaseGt in G. thermopakistaniensis. ASNaseGt exhibited optimal activity at 52 °C and pH 9.5. There was a > 3-fold increase in activity in the presence of β-mercaptoethanol. Apparent Vmax and Km values were 2735 U/mg and 0.35 mM, respectively. ASNaseGt displayed high thermostability with >80 % residual activity even after 6 h of incubation at 55 °C. Recombinant ASNaseGt existed in oligomeric form. Addition of β-mercaptoethanol lowered the degree of oligomerization and displayed that tetrameric form was the most active, with a specific activity of 4300 U/mg. Under physiological conditions, ASNaseGt displayed >50 % of the optimal activity. Localization studies in G. thermopakistaniensis revealed that ASNaseGt is a cytosolic protein. Structural and functional characterization, and localization in G. thermopakistaniensis displayed that ASNaseGt is not a type II but a type I L-asparaginase.</description><subject>asparaginase</subject><subject>Asparaginase - chemistry</subject><subject>Enzyme Stability</subject><subject>G. thermopakistaniensis</subject><subject>Geobacillus</subject><subject>Geobacillus - genetics</subject><subject>Geobacillus - metabolism</subject><subject>L-Asparaginase</subject><subject>Mercaptoethanol</subject><subject>nucleotide sequences</subject><subject>Oligomerization</subject><subject>recombinant proteins</subject><subject>Recombinant Proteins - genetics</subject><subject>Structural analysis</subject><subject>thermal stability</subject><subject>Thermostability</subject><issn>0141-8130</issn><issn>1879-0003</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2024</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkc1u2zAQhIkgQeOkfYVAx1zk8E8KdWtgpG4BAz00PRbEilomdCTR5UoF_PZlaqdXn0guv50lZxi7EXwpuKjvtsuwbUMcwC0ll3opFNfKnLGFMPdNyTlX52zBhRalyVeX7Ipom6t1JcwHdqmM5qa6bxbs148pzW6aE_QFjF3h59FNIY7_jtDvCamIPu-LTQm0gwTPYQTCwqc4FGuMLbjQ9zMV0wumIe7gNdAEY8CRAn1kFx56wk_H9Zr9_PL4tPpabr6vv60eNqVT2kxl3QE0Vds4XTeyrbRUNa_Qcy987aTpZK2N0KZrq9bnh2uuWp-_6QARhOZCXbPbg-4uxd8z0mSHQA77HkaMM1klqixpsspJVDZKZie5rDJaH1CXIlFCb3cpDJD2VnD7loLd2vcU7FsK9pBCbrw5zpjbAbv_be-2Z-DzAcBsyp-AyZLLjjnsQkI32S6GUzP-AlwEnIs</recordid><startdate>202404</startdate><enddate>202404</enddate><creator>Sania, Ayesha</creator><creator>Muhammad, Majida Atta</creator><creator>Sajed, Muhammad</creator><creator>Azim, Naseema</creator><creator>Ahmad, Nasir</creator><creator>Aslam, Mehwish</creator><creator>Tang, Xiao-Feng</creator><creator>Rashid, Naeem</creator><general>Elsevier B.V</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>7S9</scope><scope>L.6</scope></search><sort><creationdate>202404</creationdate><title>Structural and functional analyses of an L-asparaginase from Geobacillus thermopakistaniensis</title><author>Sania, Ayesha ; Muhammad, Majida Atta ; Sajed, Muhammad ; Azim, Naseema ; Ahmad, Nasir ; Aslam, Mehwish ; Tang, Xiao-Feng ; Rashid, Naeem</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c348t-6daa95b9c4692b5423605ef0f1f6c28d2648148db5bf408403bf879caeea14013</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2024</creationdate><topic>asparaginase</topic><topic>Asparaginase - chemistry</topic><topic>Enzyme Stability</topic><topic>G. thermopakistaniensis</topic><topic>Geobacillus</topic><topic>Geobacillus - genetics</topic><topic>Geobacillus - metabolism</topic><topic>L-Asparaginase</topic><topic>Mercaptoethanol</topic><topic>nucleotide sequences</topic><topic>Oligomerization</topic><topic>recombinant proteins</topic><topic>Recombinant Proteins - genetics</topic><topic>Structural analysis</topic><topic>thermal stability</topic><topic>Thermostability</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Sania, Ayesha</creatorcontrib><creatorcontrib>Muhammad, Majida Atta</creatorcontrib><creatorcontrib>Sajed, Muhammad</creatorcontrib><creatorcontrib>Azim, Naseema</creatorcontrib><creatorcontrib>Ahmad, Nasir</creatorcontrib><creatorcontrib>Aslam, Mehwish</creatorcontrib><creatorcontrib>Tang, Xiao-Feng</creatorcontrib><creatorcontrib>Rashid, Naeem</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>AGRICOLA</collection><collection>AGRICOLA - Academic</collection><jtitle>International journal of biological macromolecules</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Sania, Ayesha</au><au>Muhammad, Majida Atta</au><au>Sajed, Muhammad</au><au>Azim, Naseema</au><au>Ahmad, Nasir</au><au>Aslam, Mehwish</au><au>Tang, Xiao-Feng</au><au>Rashid, Naeem</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Structural and functional analyses of an L-asparaginase from Geobacillus thermopakistaniensis</atitle><jtitle>International journal of biological macromolecules</jtitle><addtitle>Int J Biol Macromol</addtitle><date>2024-04</date><risdate>2024</risdate><volume>263</volume><issue>Pt 2</issue><spage>130438</spage><epage>130438</epage><pages>130438-130438</pages><artnum>130438</artnum><issn>0141-8130</issn><eissn>1879-0003</eissn><abstract>Genome sequence of Geobacillus thermopakistaniensis contains an open reading frame annotated as a type II L-asparaginase (ASNaseGt). Critical structural analysis disclosed that ASNaseGt might be a type I L-asparaginase. In order to determine whether it is a type I or type II L-asparaginase, we have performed the structural-functional characterization of the recombinant protein as well as analyzed the localization of ASNaseGt in G. thermopakistaniensis. ASNaseGt exhibited optimal activity at 52 °C and pH 9.5. There was a > 3-fold increase in activity in the presence of β-mercaptoethanol. Apparent Vmax and Km values were 2735 U/mg and 0.35 mM, respectively. ASNaseGt displayed high thermostability with >80 % residual activity even after 6 h of incubation at 55 °C. Recombinant ASNaseGt existed in oligomeric form. Addition of β-mercaptoethanol lowered the degree of oligomerization and displayed that tetrameric form was the most active, with a specific activity of 4300 U/mg. Under physiological conditions, ASNaseGt displayed >50 % of the optimal activity. Localization studies in G. thermopakistaniensis revealed that ASNaseGt is a cytosolic protein. Structural and functional characterization, and localization in G. thermopakistaniensis displayed that ASNaseGt is not a type II but a type I L-asparaginase.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>38408579</pmid><doi>10.1016/j.ijbiomac.2024.130438</doi><tpages>1</tpages></addata></record> |
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| subjects | asparaginase Asparaginase - chemistry Enzyme Stability G. thermopakistaniensis Geobacillus Geobacillus - genetics Geobacillus - metabolism L-Asparaginase Mercaptoethanol nucleotide sequences Oligomerization recombinant proteins Recombinant Proteins - genetics Structural analysis thermal stability Thermostability |
| title | Structural and functional analyses of an L-asparaginase from Geobacillus thermopakistaniensis |
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