An Acid-Adapted Endo-[alpha]-1,5-l-arabinanase for Pectin Releasing
An arabinanase gene was cloned by overlap-PCR from Penicillium sp. Y702 and expressed in Pichia pastoris. The recombinant enzyme was named AbnC702 with 20 U/mg of endo-arabinanase activity toward linear [alpha]-1,5-l-arabinan. The optimal pH and temperature of AbnC702 were 5.0 and 50 °C, respectivel...
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Veröffentlicht in: | Applied biochemistry and biotechnology 2016-11, Vol.180 (5), p.900 |
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description | An arabinanase gene was cloned by overlap-PCR from Penicillium sp. Y702 and expressed in Pichia pastoris. The recombinant enzyme was named AbnC702 with 20 U/mg of endo-arabinanase activity toward linear [alpha]-1,5-l-arabinan. The optimal pH and temperature of AbnC702 were 5.0 and 50 °C, respectively. The recombinant AbnC702 was highly stable at pH 5.0-7.0 and 50 °C. It could retain about 72.3 % of maximum specific activity at pH 5.0 after incubation for 2.5 h, which indicated AbnC702 was an acid-adapted enzyme. The K m and V max values were 24.8±4.7 mg/ml and 88.5±5.6 U/mg, respectively. A three-dimensional structure of AbnC702 was made by homology modeling, and the counting of acidic/basic amino residues within the region of 10 Å around the active site, as well the hydrogen bonds within the area of 5 Å around the active site, might theoretically interpret the acid adaptability of AbnC702. Analysis of hydrolysis products by thin layer chromatography (TLC) combined with high-performance liquid chromatography (HPLC) verified that the recombinant AbnC702 was an endo-1,5-[alpha]-l-arabinanase, which yielded arabinobiose and arabinotriose as major products. AbnC702 was applied in pectin extraction from apple pomace with synergistic action of [alpha]-L-arabinofuranosidase. |
doi_str_mv | 10.1007/s12010-016-2141-5 |
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Y702 and expressed in Pichia pastoris. The recombinant enzyme was named AbnC702 with 20 U/mg of endo-arabinanase activity toward linear [alpha]-1,5-l-arabinan. The optimal pH and temperature of AbnC702 were 5.0 and 50 °C, respectively. The recombinant AbnC702 was highly stable at pH 5.0-7.0 and 50 °C. It could retain about 72.3 % of maximum specific activity at pH 5.0 after incubation for 2.5 h, which indicated AbnC702 was an acid-adapted enzyme. The K m and V max values were 24.8±4.7 mg/ml and 88.5±5.6 U/mg, respectively. A three-dimensional structure of AbnC702 was made by homology modeling, and the counting of acidic/basic amino residues within the region of 10 Å around the active site, as well the hydrogen bonds within the area of 5 Å around the active site, might theoretically interpret the acid adaptability of AbnC702. Analysis of hydrolysis products by thin layer chromatography (TLC) combined with high-performance liquid chromatography (HPLC) verified that the recombinant AbnC702 was an endo-1,5-[alpha]-l-arabinanase, which yielded arabinobiose and arabinotriose as major products. AbnC702 was applied in pectin extraction from apple pomace with synergistic action of [alpha]-L-arabinofuranosidase.</description><identifier>ISSN: 0273-2289</identifier><identifier>EISSN: 1559-0291</identifier><identifier>DOI: 10.1007/s12010-016-2141-5</identifier><language>eng</language><publisher>Totowa: Springer Nature B.V</publisher><subject>Adaptability ; Apples ; Biotechnology ; Enzymes ; Extraction processes ; Liquid chromatography ; Pectin ; Thin layer chromatography</subject><ispartof>Applied biochemistry and biotechnology, 2016-11, Vol.180 (5), p.900</ispartof><rights>Springer Science+Business Media New York 2016</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids></links><search><creatorcontrib>Lang, Chong</creatorcontrib><creatorcontrib>Yang, Rujian</creatorcontrib><creatorcontrib>Yang, Ying</creatorcontrib><creatorcontrib>Gao, Bei</creatorcontrib><creatorcontrib>Zhao, Li</creatorcontrib><creatorcontrib>Wei, Wei</creatorcontrib><creatorcontrib>Wang, Hualei</creatorcontrib><creatorcontrib>Matsukawa, Shingo</creatorcontrib><creatorcontrib>Xie, Jingli</creatorcontrib><creatorcontrib>Wei, Dongzhi</creatorcontrib><title>An Acid-Adapted Endo-[alpha]-1,5-l-arabinanase for Pectin Releasing</title><title>Applied biochemistry and biotechnology</title><description>An arabinanase gene was cloned by overlap-PCR from Penicillium sp. Y702 and expressed in Pichia pastoris. The recombinant enzyme was named AbnC702 with 20 U/mg of endo-arabinanase activity toward linear [alpha]-1,5-l-arabinan. The optimal pH and temperature of AbnC702 were 5.0 and 50 °C, respectively. The recombinant AbnC702 was highly stable at pH 5.0-7.0 and 50 °C. It could retain about 72.3 % of maximum specific activity at pH 5.0 after incubation for 2.5 h, which indicated AbnC702 was an acid-adapted enzyme. The K m and V max values were 24.8±4.7 mg/ml and 88.5±5.6 U/mg, respectively. A three-dimensional structure of AbnC702 was made by homology modeling, and the counting of acidic/basic amino residues within the region of 10 Å around the active site, as well the hydrogen bonds within the area of 5 Å around the active site, might theoretically interpret the acid adaptability of AbnC702. Analysis of hydrolysis products by thin layer chromatography (TLC) combined with high-performance liquid chromatography (HPLC) verified that the recombinant AbnC702 was an endo-1,5-[alpha]-l-arabinanase, which yielded arabinobiose and arabinotriose as major products. AbnC702 was applied in pectin extraction from apple pomace with synergistic action of [alpha]-L-arabinofuranosidase.</description><subject>Adaptability</subject><subject>Apples</subject><subject>Biotechnology</subject><subject>Enzymes</subject><subject>Extraction processes</subject><subject>Liquid chromatography</subject><subject>Pectin</subject><subject>Thin layer chromatography</subject><issn>0273-2289</issn><issn>1559-0291</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNotjktLw0AURgdRMFZ_gLuAW6_eO8mdZJah1AcUFOlOpMwrNSVMYib9_wZ0853dOZ8Qt4QPhFg9JpJICEgKJJUEfCYyYtaAUtO5yFBWBUhZ60txldIRkWTNVSbWTcwb13lovBnn4PNN9AN8mn78Nl9A9ww9mMnYLppoUsjbYcrfg5u7mH-EPpjUxcO1uGhNn8LNP1di97TZrV9g-_b8um62cFDLEe9Z6ULZSrFka02QznJtvHZSOyarStZolS99CK1E1VayVpqp9OQ82lCsxN2fdpyGn1NI8_44nKa4FPdUF6yXWQK_lCRKow</recordid><startdate>20161101</startdate><enddate>20161101</enddate><creator>Lang, Chong</creator><creator>Yang, Rujian</creator><creator>Yang, Ying</creator><creator>Gao, Bei</creator><creator>Zhao, Li</creator><creator>Wei, Wei</creator><creator>Wang, Hualei</creator><creator>Matsukawa, Shingo</creator><creator>Xie, Jingli</creator><creator>Wei, Dongzhi</creator><general>Springer Nature B.V</general><scope>3V.</scope><scope>7ST</scope><scope>7T7</scope><scope>7TM</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>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</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>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7P</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>RC3</scope><scope>SOI</scope></search><sort><creationdate>20161101</creationdate><title>An Acid-Adapted Endo-[alpha]-1,5-l-arabinanase for Pectin Releasing</title><author>Lang, Chong ; Yang, Rujian ; Yang, Ying ; Gao, Bei ; Zhao, Li ; Wei, Wei ; Wang, Hualei ; Matsukawa, Shingo ; Xie, Jingli ; Wei, Dongzhi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-g659-dd56936b76525bbae2cb58ad9c29c51b64590b6d4deef206f72869514d1cd0be3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>Adaptability</topic><topic>Apples</topic><topic>Biotechnology</topic><topic>Enzymes</topic><topic>Extraction processes</topic><topic>Liquid chromatography</topic><topic>Pectin</topic><topic>Thin layer chromatography</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Lang, Chong</creatorcontrib><creatorcontrib>Yang, Rujian</creatorcontrib><creatorcontrib>Yang, Ying</creatorcontrib><creatorcontrib>Gao, Bei</creatorcontrib><creatorcontrib>Zhao, Li</creatorcontrib><creatorcontrib>Wei, Wei</creatorcontrib><creatorcontrib>Wang, Hualei</creatorcontrib><creatorcontrib>Matsukawa, Shingo</creatorcontrib><creatorcontrib>Xie, Jingli</creatorcontrib><creatorcontrib>Wei, Dongzhi</creatorcontrib><collection>ProQuest Central (Corporate)</collection><collection>Environment Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Nucleic Acids Abstracts</collection><collection>ProQuest Health and Medical</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Science Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>ProQuest Science Journals</collection><collection>ProQuest Biological Science Journals</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central Basic</collection><collection>Genetics Abstracts</collection><collection>Environment Abstracts</collection><jtitle>Applied biochemistry and biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Lang, Chong</au><au>Yang, Rujian</au><au>Yang, Ying</au><au>Gao, Bei</au><au>Zhao, Li</au><au>Wei, Wei</au><au>Wang, Hualei</au><au>Matsukawa, Shingo</au><au>Xie, Jingli</au><au>Wei, Dongzhi</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>An Acid-Adapted Endo-[alpha]-1,5-l-arabinanase for Pectin Releasing</atitle><jtitle>Applied biochemistry and biotechnology</jtitle><date>2016-11-01</date><risdate>2016</risdate><volume>180</volume><issue>5</issue><spage>900</spage><pages>900-</pages><issn>0273-2289</issn><eissn>1559-0291</eissn><abstract>An arabinanase gene was cloned by overlap-PCR from Penicillium sp. Y702 and expressed in Pichia pastoris. The recombinant enzyme was named AbnC702 with 20 U/mg of endo-arabinanase activity toward linear [alpha]-1,5-l-arabinan. The optimal pH and temperature of AbnC702 were 5.0 and 50 °C, respectively. The recombinant AbnC702 was highly stable at pH 5.0-7.0 and 50 °C. It could retain about 72.3 % of maximum specific activity at pH 5.0 after incubation for 2.5 h, which indicated AbnC702 was an acid-adapted enzyme. The K m and V max values were 24.8±4.7 mg/ml and 88.5±5.6 U/mg, respectively. A three-dimensional structure of AbnC702 was made by homology modeling, and the counting of acidic/basic amino residues within the region of 10 Å around the active site, as well the hydrogen bonds within the area of 5 Å around the active site, might theoretically interpret the acid adaptability of AbnC702. Analysis of hydrolysis products by thin layer chromatography (TLC) combined with high-performance liquid chromatography (HPLC) verified that the recombinant AbnC702 was an endo-1,5-[alpha]-l-arabinanase, which yielded arabinobiose and arabinotriose as major products. AbnC702 was applied in pectin extraction from apple pomace with synergistic action of [alpha]-L-arabinofuranosidase.</abstract><cop>Totowa</cop><pub>Springer Nature B.V</pub><doi>10.1007/s12010-016-2141-5</doi></addata></record> |
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subjects | Adaptability Apples Biotechnology Enzymes Extraction processes Liquid chromatography Pectin Thin layer chromatography |
title | An Acid-Adapted Endo-[alpha]-1,5-l-arabinanase for Pectin Releasing |
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