Deleting the Ig-Like Domain of Alicyclobacillus acidocaldarius Endoglucanase Cel9A Causes a Simultaneous Increase in the Activity and Stability
Endoglucanase Cel9A from Alicyclobacillus acidocaldarius (AaCel9A) is a monomeric enzyme with 537 residues. This enzyme has an Ig-like domain in the N-terminus of the catalytic domain. In this study, the role of the Ig-like domain on the activity, stability, and structural rigidity of AaCel9A and th...
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| Veröffentlicht in: | Molecular biotechnology 2016-01, Vol.58 (1), p.12-21 |
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| creator | Younesi, Fereshteh S. Pazhang, Mohammad Najavand, Saeed Rahimizadeh, Parastou Akbarian, Mohsen Mohammadian, Mehdi Khajeh, Khosro |
| description | Endoglucanase Cel9A from
Alicyclobacillus acidocaldarius
(AaCel9A) is a monomeric enzyme with 537 residues. This enzyme has an Ig-like domain in the N-terminus of the catalytic domain. In this study, the role of the Ig-like domain on the activity, stability, and structural rigidity of AaCel9A and the effect of calcium on enzyme activity and stability were examined by comparing a truncated enzyme with deletion of the Ig-like domain (AaCel9AΔN) to the wild-type enzyme. Our results showed that the deletion of the Ig-like domain increased the catalytic efficiency of the truncated enzyme up to threefold without any significant changes in the
K
m
of the enzyme. Furthermore, pH and temperature optimum for activity were shifted from 6.5 to 7.5 and from 65 to 60 °C, respectively, by deletion of the Ig-like domain. The thermal stability and fluorescence quenching results indicated that the stability and rigidity of the truncated enzyme have been more than that of the wild-type enzyme. Calcium similarly increased the catalytic efficiency of the enzymes (up to 40 %) and remarkably raised the stability of the AaCel9A compared to the AaCel9AΔN. This shows that Ig-like domain has a role in the increase of the enzyme stability by calcium in the wild-type enzyme. |
| doi_str_mv | 10.1007/s12033-015-9900-3 |
| format | Article |
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Alicyclobacillus acidocaldarius
(AaCel9A) is a monomeric enzyme with 537 residues. This enzyme has an Ig-like domain in the N-terminus of the catalytic domain. In this study, the role of the Ig-like domain on the activity, stability, and structural rigidity of AaCel9A and the effect of calcium on enzyme activity and stability were examined by comparing a truncated enzyme with deletion of the Ig-like domain (AaCel9AΔN) to the wild-type enzyme. Our results showed that the deletion of the Ig-like domain increased the catalytic efficiency of the truncated enzyme up to threefold without any significant changes in the
K
m
of the enzyme. Furthermore, pH and temperature optimum for activity were shifted from 6.5 to 7.5 and from 65 to 60 °C, respectively, by deletion of the Ig-like domain. The thermal stability and fluorescence quenching results indicated that the stability and rigidity of the truncated enzyme have been more than that of the wild-type enzyme. Calcium similarly increased the catalytic efficiency of the enzymes (up to 40 %) and remarkably raised the stability of the AaCel9A compared to the AaCel9AΔN. This shows that Ig-like domain has a role in the increase of the enzyme stability by calcium in the wild-type enzyme.</description><identifier>ISSN: 1073-6085</identifier><identifier>EISSN: 1559-0305</identifier><identifier>DOI: 10.1007/s12033-015-9900-3</identifier><identifier>PMID: 26537871</identifier><language>eng</language><publisher>New York: Springer US</publisher><subject>Alicyclobacillus - chemistry ; Alicyclobacillus - enzymology ; Alicyclobacillus acidocaldarius ; Amino Acid Sequence - genetics ; Bacteria ; Biocatalysts ; Biochemistry ; Biological Techniques ; Biotechnology ; Calcium ; Calcium - metabolism ; Catalytic Domain ; Cell Biology ; Cellulase - chemistry ; Cellulase - genetics ; Cellulase - metabolism ; Chemistry ; Chemistry and Materials Science ; Enzymatic activity ; Enzyme Stability ; Enzymes ; Human Genetics ; Immunoglobulins ; Protein Science ; Rigidity ; Sequence Deletion ; Substrate Specificity</subject><ispartof>Molecular biotechnology, 2016-01, Vol.58 (1), p.12-21</ispartof><rights>Springer Science+Business Media New York 2015</rights><rights>Springer Science+Business Media New York 2016</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c475t-74ccba83338bd1bf6cfb9187dcf5d308e6ec288bdee349ce88f992117a3020083</citedby><cites>FETCH-LOGICAL-c475t-74ccba83338bd1bf6cfb9187dcf5d308e6ec288bdee349ce88f992117a3020083</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s12033-015-9900-3$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s12033-015-9900-3$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,777,781,27905,27906,41469,42538,51300</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26537871$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Younesi, Fereshteh S.</creatorcontrib><creatorcontrib>Pazhang, Mohammad</creatorcontrib><creatorcontrib>Najavand, Saeed</creatorcontrib><creatorcontrib>Rahimizadeh, Parastou</creatorcontrib><creatorcontrib>Akbarian, Mohsen</creatorcontrib><creatorcontrib>Mohammadian, Mehdi</creatorcontrib><creatorcontrib>Khajeh, Khosro</creatorcontrib><title>Deleting the Ig-Like Domain of Alicyclobacillus acidocaldarius Endoglucanase Cel9A Causes a Simultaneous Increase in the Activity and Stability</title><title>Molecular biotechnology</title><addtitle>Mol Biotechnol</addtitle><addtitle>Mol Biotechnol</addtitle><description>Endoglucanase Cel9A from
Alicyclobacillus acidocaldarius
(AaCel9A) is a monomeric enzyme with 537 residues. This enzyme has an Ig-like domain in the N-terminus of the catalytic domain. In this study, the role of the Ig-like domain on the activity, stability, and structural rigidity of AaCel9A and the effect of calcium on enzyme activity and stability were examined by comparing a truncated enzyme with deletion of the Ig-like domain (AaCel9AΔN) to the wild-type enzyme. Our results showed that the deletion of the Ig-like domain increased the catalytic efficiency of the truncated enzyme up to threefold without any significant changes in the
K
m
of the enzyme. Furthermore, pH and temperature optimum for activity were shifted from 6.5 to 7.5 and from 65 to 60 °C, respectively, by deletion of the Ig-like domain. The thermal stability and fluorescence quenching results indicated that the stability and rigidity of the truncated enzyme have been more than that of the wild-type enzyme. Calcium similarly increased the catalytic efficiency of the enzymes (up to 40 %) and remarkably raised the stability of the AaCel9A compared to the AaCel9AΔN. This shows that Ig-like domain has a role in the increase of the enzyme stability by calcium in the wild-type enzyme.</description><subject>Alicyclobacillus - chemistry</subject><subject>Alicyclobacillus - enzymology</subject><subject>Alicyclobacillus acidocaldarius</subject><subject>Amino Acid Sequence - genetics</subject><subject>Bacteria</subject><subject>Biocatalysts</subject><subject>Biochemistry</subject><subject>Biological Techniques</subject><subject>Biotechnology</subject><subject>Calcium</subject><subject>Calcium - metabolism</subject><subject>Catalytic Domain</subject><subject>Cell Biology</subject><subject>Cellulase - chemistry</subject><subject>Cellulase - genetics</subject><subject>Cellulase - metabolism</subject><subject>Chemistry</subject><subject>Chemistry and Materials Science</subject><subject>Enzymatic activity</subject><subject>Enzyme Stability</subject><subject>Enzymes</subject><subject>Human Genetics</subject><subject>Immunoglobulins</subject><subject>Protein Science</subject><subject>Rigidity</subject><subject>Sequence Deletion</subject><subject>Substrate Specificity</subject><issn>1073-6085</issn><issn>1559-0305</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNqNkU1v1DAQhiNERT_gB3BBlrhwCR3HcWwfV9tCV1qph8LZcpzJ4uLExU4q7a_gL-MoBSEkpJ7Go3n8zsdbFG8pfKQA4jLRChgrgfJSKYCSvSjOKOeqBAb8ZX6DYGUDkp8W5yndA1SU1-xVcVo1nAkp6Fnx8wo9Tm48kOkbkt2h3LvvSK7CYNxIQk823tmj9aE11nk_J5JjF6zxnYkup9djFw5-tmY0CckWvdqQrZkTZpLcuWH2kxkxZHI32ogLlIWXXhs7uUc3HYkZO3I3mdb5nL0uTnrjE755ihfF10_XX7Y35f7282672Ze2FnwqRW1tayRjTLYdbfvG9q2iUnS25x0DiQ3aSuYaIquVRSl7pSpKhWFQAUh2UXxYdR9i-DFjmvTgkkXv12k1FQ1XsmZN_Rw0n1hVimX0_T_ofZjjmBfJFOdCMkohU3SlbAwpRez1Q3SDiUdNQS_G6tVYnY3Vi7F6UX73pDy3A3Z_fvx2MgPVCqRcGg8Y_2r9X9Vf3EiuXw</recordid><startdate>20160101</startdate><enddate>20160101</enddate><creator>Younesi, Fereshteh S.</creator><creator>Pazhang, Mohammad</creator><creator>Najavand, Saeed</creator><creator>Rahimizadeh, Parastou</creator><creator>Akbarian, Mohsen</creator><creator>Mohammadian, Mehdi</creator><creator>Khajeh, Khosro</creator><general>Springer US</general><general>Springer Nature 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>3V.</scope><scope>7QL</scope><scope>7QO</scope><scope>7T7</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PTHSS</scope><scope>Q9U</scope><scope>7X8</scope></search><sort><creationdate>20160101</creationdate><title>Deleting the Ig-Like Domain of Alicyclobacillus acidocaldarius Endoglucanase Cel9A Causes a Simultaneous Increase in the Activity and Stability</title><author>Younesi, Fereshteh S. ; Pazhang, Mohammad ; Najavand, Saeed ; Rahimizadeh, Parastou ; Akbarian, Mohsen ; Mohammadian, Mehdi ; Khajeh, Khosro</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c475t-74ccba83338bd1bf6cfb9187dcf5d308e6ec288bdee349ce88f992117a3020083</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>Alicyclobacillus - 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Academic</collection><jtitle>Molecular biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Younesi, Fereshteh S.</au><au>Pazhang, Mohammad</au><au>Najavand, Saeed</au><au>Rahimizadeh, Parastou</au><au>Akbarian, Mohsen</au><au>Mohammadian, Mehdi</au><au>Khajeh, Khosro</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Deleting the Ig-Like Domain of Alicyclobacillus acidocaldarius Endoglucanase Cel9A Causes a Simultaneous Increase in the Activity and Stability</atitle><jtitle>Molecular biotechnology</jtitle><stitle>Mol Biotechnol</stitle><addtitle>Mol Biotechnol</addtitle><date>2016-01-01</date><risdate>2016</risdate><volume>58</volume><issue>1</issue><spage>12</spage><epage>21</epage><pages>12-21</pages><issn>1073-6085</issn><eissn>1559-0305</eissn><abstract>Endoglucanase Cel9A from
Alicyclobacillus acidocaldarius
(AaCel9A) is a monomeric enzyme with 537 residues. This enzyme has an Ig-like domain in the N-terminus of the catalytic domain. In this study, the role of the Ig-like domain on the activity, stability, and structural rigidity of AaCel9A and the effect of calcium on enzyme activity and stability were examined by comparing a truncated enzyme with deletion of the Ig-like domain (AaCel9AΔN) to the wild-type enzyme. Our results showed that the deletion of the Ig-like domain increased the catalytic efficiency of the truncated enzyme up to threefold without any significant changes in the
K
m
of the enzyme. Furthermore, pH and temperature optimum for activity were shifted from 6.5 to 7.5 and from 65 to 60 °C, respectively, by deletion of the Ig-like domain. The thermal stability and fluorescence quenching results indicated that the stability and rigidity of the truncated enzyme have been more than that of the wild-type enzyme. Calcium similarly increased the catalytic efficiency of the enzymes (up to 40 %) and remarkably raised the stability of the AaCel9A compared to the AaCel9AΔN. This shows that Ig-like domain has a role in the increase of the enzyme stability by calcium in the wild-type enzyme.</abstract><cop>New York</cop><pub>Springer US</pub><pmid>26537871</pmid><doi>10.1007/s12033-015-9900-3</doi><tpages>10</tpages></addata></record> |
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| ispartof | Molecular biotechnology, 2016-01, Vol.58 (1), p.12-21 |
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| subjects | Alicyclobacillus - chemistry Alicyclobacillus - enzymology Alicyclobacillus acidocaldarius Amino Acid Sequence - genetics Bacteria Biocatalysts Biochemistry Biological Techniques Biotechnology Calcium Calcium - metabolism Catalytic Domain Cell Biology Cellulase - chemistry Cellulase - genetics Cellulase - metabolism Chemistry Chemistry and Materials Science Enzymatic activity Enzyme Stability Enzymes Human Genetics Immunoglobulins Protein Science Rigidity Sequence Deletion Substrate Specificity |
| title | Deleting the Ig-Like Domain of Alicyclobacillus acidocaldarius Endoglucanase Cel9A Causes a Simultaneous Increase in the Activity and Stability |
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