Introduction of additional thiol groups into glucoamylase in Aspergillus Awamori and their effect on the thermal stability and catalytic activity of the enzyme
Five mutant forms of glucoamylase (GA) from the filamentous fungus Aspergillus awamori with artificial disulfide bonds (4D-G137A\A14C, 6D-A14C\Y419C\G137A, 10D-V13C\G396C, 11D-V13C\G396C\A14C\Y419C\G137A, and 20D-G137A\A246C\A14C) were constructed using molecular modeling simulations and experimenta...
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| Veröffentlicht in: | Applied biochemistry and microbiology 2014-03, Vol.50 (2), p.118-124 |
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| creator | Surzhik, M. A. Schmidt, A. E. Glazunov, E. A. Firsov, D. L. Petukhov, M. G. |
| description | Five mutant forms of glucoamylase (GA) from the filamentous fungus
Aspergillus awamori
with artificial disulfide bonds (4D-G137A\A14C, 6D-A14C\Y419C\G137A, 10D-V13C\G396C, 11D-V13C\G396C\A14C\Y419C\G137A, and 20D-G137A\A246C\A14C) were constructed using molecular modeling simulations and experimentally tested for thermostability. The introduction of two additional disulfide bonds between its first and thirteenth α-helices and that of the loop located close to a catalytic residue—E400—made it possible to assess the effects of disulfide bridges on protein thermostability. The mutant proteins with combined amino acid substitutions G137A\A14C, V13C\G396C\A14C\Y419C\G137A, and G137A\A246C\A14C showed higher thermal stability as compared to the wild-type protein. At the same time, new disulfide bridges in the mutant A14C\Y419C\G137A and V13C\G396C proteins led to the destabilization of their structure and the loss of thermal stability. |
| doi_str_mv | 10.1134/S0003683814020185 |
| format | Article |
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Aspergillus awamori
with artificial disulfide bonds (4D-G137A\A14C, 6D-A14C\Y419C\G137A, 10D-V13C\G396C, 11D-V13C\G396C\A14C\Y419C\G137A, and 20D-G137A\A246C\A14C) were constructed using molecular modeling simulations and experimentally tested for thermostability. The introduction of two additional disulfide bonds between its first and thirteenth α-helices and that of the loop located close to a catalytic residue—E400—made it possible to assess the effects of disulfide bridges on protein thermostability. The mutant proteins with combined amino acid substitutions G137A\A14C, V13C\G396C\A14C\Y419C\G137A, and G137A\A246C\A14C showed higher thermal stability as compared to the wild-type protein. At the same time, new disulfide bridges in the mutant A14C\Y419C\G137A and V13C\G396C proteins led to the destabilization of their structure and the loss of thermal stability.</description><identifier>ISSN: 0003-6838</identifier><identifier>EISSN: 1608-3024</identifier><identifier>DOI: 10.1134/S0003683814020185</identifier><language>eng</language><publisher>Boston: Springer US</publisher><subject>Amino acids ; Aspergillus awamori ; Biocatalysts ; Biochemistry ; Biomedical and Life Sciences ; Biophysics ; Enzymes ; Fungi ; Life Sciences ; Medical Microbiology ; Microbiology ; Proteins</subject><ispartof>Applied biochemistry and microbiology, 2014-03, Vol.50 (2), p.118-124</ispartof><rights>Pleiades Publishing, Inc. 2014</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c349t-58bbe28fcda0e4258e1e24c3d6cb0c1150196ec6a38b9fb9ccf43990e03b14393</citedby><cites>FETCH-LOGICAL-c349t-58bbe28fcda0e4258e1e24c3d6cb0c1150196ec6a38b9fb9ccf43990e03b14393</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1134/S0003683814020185$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1134/S0003683814020185$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,776,780,27901,27902,41464,42533,51294</link.rule.ids></links><search><creatorcontrib>Surzhik, M. A.</creatorcontrib><creatorcontrib>Schmidt, A. E.</creatorcontrib><creatorcontrib>Glazunov, E. A.</creatorcontrib><creatorcontrib>Firsov, D. L.</creatorcontrib><creatorcontrib>Petukhov, M. G.</creatorcontrib><title>Introduction of additional thiol groups into glucoamylase in Aspergillus Awamori and their effect on the thermal stability and catalytic activity of the enzyme</title><title>Applied biochemistry and microbiology</title><addtitle>Appl Biochem Microbiol</addtitle><description>Five mutant forms of glucoamylase (GA) from the filamentous fungus
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with artificial disulfide bonds (4D-G137A\A14C, 6D-A14C\Y419C\G137A, 10D-V13C\G396C, 11D-V13C\G396C\A14C\Y419C\G137A, and 20D-G137A\A246C\A14C) were constructed using molecular modeling simulations and experimentally tested for thermostability. The introduction of two additional disulfide bonds between its first and thirteenth α-helices and that of the loop located close to a catalytic residue—E400—made it possible to assess the effects of disulfide bridges on protein thermostability. The mutant proteins with combined amino acid substitutions G137A\A14C, V13C\G396C\A14C\Y419C\G137A, and G137A\A246C\A14C showed higher thermal stability as compared to the wild-type protein. At the same time, new disulfide bridges in the mutant A14C\Y419C\G137A and V13C\G396C proteins led to the destabilization of their structure and the loss of thermal stability.</description><subject>Amino acids</subject><subject>Aspergillus awamori</subject><subject>Biocatalysts</subject><subject>Biochemistry</subject><subject>Biomedical and Life Sciences</subject><subject>Biophysics</subject><subject>Enzymes</subject><subject>Fungi</subject><subject>Life Sciences</subject><subject>Medical Microbiology</subject><subject>Microbiology</subject><subject>Proteins</subject><issn>0003-6838</issn><issn>1608-3024</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>BENPR</sourceid><recordid>eNp1kctKxDAUhoMoOF4ewF3AjZtq0qSdZDmIlwHBhbouaXo6ZkibMUmV-jK-qqnjQhQXIefy_T-HcxA6oeScUsYvHgghrBRMUE5yQkWxg2a0JCJjJOe7aDa1s6m_jw5CWKdUlkLO0Meyj941g47G9di1WDWNmWJlcXw2zuKVd8MmYNNHh1d20E51o1UBUgUvwgb8ylg7BLx4U53zBqu-SUowHkPbgo44-aZ8er5LriGq2lgTxy9Sq6jsGI3GKo3wOpXTEBMP_fvYwRHaa5UNcPz9H6Kn66vHy9vs7v5mebm4yzTjMmaFqGvIRasbRYDnhQAKOdesKXVNNKUFobIEXSomatnWUuuWMykJEFbTFLFDdLb13Xj3MkCIVWeCBmtVD24IVXIQ85JyThJ6-gtdu8GnhX1RnBWyZPNE0S2lvQvBQ1ttvOmUHytKqulk1Z-TJU2-1YTE9ivwP5z_FX0C8jqbZg</recordid><startdate>20140301</startdate><enddate>20140301</enddate><creator>Surzhik, M. 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A.</au><au>Schmidt, A. E.</au><au>Glazunov, E. A.</au><au>Firsov, D. L.</au><au>Petukhov, M. G.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Introduction of additional thiol groups into glucoamylase in Aspergillus Awamori and their effect on the thermal stability and catalytic activity of the enzyme</atitle><jtitle>Applied biochemistry and microbiology</jtitle><stitle>Appl Biochem Microbiol</stitle><date>2014-03-01</date><risdate>2014</risdate><volume>50</volume><issue>2</issue><spage>118</spage><epage>124</epage><pages>118-124</pages><issn>0003-6838</issn><eissn>1608-3024</eissn><abstract>Five mutant forms of glucoamylase (GA) from the filamentous fungus
Aspergillus awamori
with artificial disulfide bonds (4D-G137A\A14C, 6D-A14C\Y419C\G137A, 10D-V13C\G396C, 11D-V13C\G396C\A14C\Y419C\G137A, and 20D-G137A\A246C\A14C) were constructed using molecular modeling simulations and experimentally tested for thermostability. The introduction of two additional disulfide bonds between its first and thirteenth α-helices and that of the loop located close to a catalytic residue—E400—made it possible to assess the effects of disulfide bridges on protein thermostability. The mutant proteins with combined amino acid substitutions G137A\A14C, V13C\G396C\A14C\Y419C\G137A, and G137A\A246C\A14C showed higher thermal stability as compared to the wild-type protein. At the same time, new disulfide bridges in the mutant A14C\Y419C\G137A and V13C\G396C proteins led to the destabilization of their structure and the loss of thermal stability.</abstract><cop>Boston</cop><pub>Springer US</pub><doi>10.1134/S0003683814020185</doi><tpages>7</tpages></addata></record> |
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| source | SpringerNature Complete Journals |
| subjects | Amino acids Aspergillus awamori Biocatalysts Biochemistry Biomedical and Life Sciences Biophysics Enzymes Fungi Life Sciences Medical Microbiology Microbiology Proteins |
| title | Introduction of additional thiol groups into glucoamylase in Aspergillus Awamori and their effect on the thermal stability and catalytic activity of the enzyme |
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