Overexpression of a fungal β-mannanase from Bispora sp. MEY-1 in maize seeds and enzyme characterization
Mannans and heteromannans are widespread in plants cell walls and are well-known as anti-nutritional factors in animal feed. To remove these factors, it is common practice to incorporate endo-β-mannanase into feed for efficient nutrition absorption. The objective of this study was to overexpress a β...
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description | Mannans and heteromannans are widespread in plants cell walls and are well-known as anti-nutritional factors in animal feed. To remove these factors, it is common practice to incorporate endo-β-mannanase into feed for efficient nutrition absorption. The objective of this study was to overexpress a β-mannanase gene directly in maize, the main ingredient of animal feed, to simplify the process of feed production.
The man5A gene encoding an excellent β-mannanase from acidophilic Bispora sp. MEY-1 was selected for heterologous overexpression. Expression of the modified gene (man5As) was driven by the embryo-specific promoter ZM-leg1A, and the transgene was transferred to three generations by backcrossing with commercial inbred Zheng58. Its exogenous integration into the maize embryonic genome and tissue specific expression in seeds were confirmed by PCR and Southern blot and Western blot analysis, respectively. Transgenic plants at BC3 generation showed agronomic traits statistically similar to Zheng58 except for less plant height (154.0 cm vs 158.3 cm). The expression level of MAN5AS reached up to 26,860 units per kilogram of maize seeds. Compared with its counterpart produced in Pichia pastoris, seed-derived MAN5AS had higher temperature optimum (90°C), and remained more β-mannanase activities after pelleting at 80°C, 100°C or 120°C.
This study shows the genetically stable overexpression of a fungal β-mannanase in maize and offers an effective and economic approach for transgene containment in maize for direct utilization without any purification or supplementation procedures. |
doi_str_mv | 10.1371/journal.pone.0056146 |
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The man5A gene encoding an excellent β-mannanase from acidophilic Bispora sp. MEY-1 was selected for heterologous overexpression. Expression of the modified gene (man5As) was driven by the embryo-specific promoter ZM-leg1A, and the transgene was transferred to three generations by backcrossing with commercial inbred Zheng58. Its exogenous integration into the maize embryonic genome and tissue specific expression in seeds were confirmed by PCR and Southern blot and Western blot analysis, respectively. Transgenic plants at BC3 generation showed agronomic traits statistically similar to Zheng58 except for less plant height (154.0 cm vs 158.3 cm). The expression level of MAN5AS reached up to 26,860 units per kilogram of maize seeds. Compared with its counterpart produced in Pichia pastoris, seed-derived MAN5AS had higher temperature optimum (90°C), and remained more β-mannanase activities after pelleting at 80°C, 100°C or 120°C.
This study shows the genetically stable overexpression of a fungal β-mannanase in maize and offers an effective and economic approach for transgene containment in maize for direct utilization without any purification or supplementation procedures.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0056146</identifier><identifier>PMID: 23409143</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Agriculture ; Agronomy ; Animal Feed ; Ascomycota - enzymology ; Ascomycota - genetics ; beta-Mannosidase - genetics ; beta-Mannosidase - metabolism ; Biology ; Biotechnology ; Cell walls ; Containment ; Corn ; Embryos ; Enzyme Activation ; Enzyme Stability ; Enzymes ; Feed additives ; Feeds ; Food additives ; Fungi ; Gene Expression ; Genetic Vectors - genetics ; Genomes ; Genomics ; Inbreeding ; Laboratories ; Mannanases ; Nutrition ; Pelleting ; Phenotype ; Pichia - genetics ; Pichia pastoris ; Purification ; Regeneration ; Seeds ; Seeds - genetics ; Seeds - physiology ; Tobacco ; Transformation, Genetic ; Transgenic plants ; Yeast ; Zea mays ; Zea mays - genetics ; Zea mays - physiology</subject><ispartof>PloS one, 2013-02, Vol.8 (2), p.e56146-e56146</ispartof><rights>2013 Xu et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: https://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2013 Xu et al 2013 Xu et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c526t-8ad586b416395dabaea7114585acec2711bcd848e69adc61eb582031e1216c963</citedby><cites>FETCH-LOGICAL-c526t-8ad586b416395dabaea7114585acec2711bcd848e69adc61eb582031e1216c963</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3569411/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3569411/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,725,778,782,862,883,2098,2917,23853,27911,27912,53778,53780,79355,79356</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23409143$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Li, Yi</contributor><creatorcontrib>Xu, Xiaolu</creatorcontrib><creatorcontrib>Zhang, Yuhong</creatorcontrib><creatorcontrib>Meng, Qingchang</creatorcontrib><creatorcontrib>Meng, Kun</creatorcontrib><creatorcontrib>Zhang, Wei</creatorcontrib><creatorcontrib>Zhou, Xiaojin</creatorcontrib><creatorcontrib>Luo, Huiying</creatorcontrib><creatorcontrib>Chen, Rumei</creatorcontrib><creatorcontrib>Yang, Peilong</creatorcontrib><creatorcontrib>Yao, Bin</creatorcontrib><title>Overexpression of a fungal β-mannanase from Bispora sp. MEY-1 in maize seeds and enzyme characterization</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Mannans and heteromannans are widespread in plants cell walls and are well-known as anti-nutritional factors in animal feed. To remove these factors, it is common practice to incorporate endo-β-mannanase into feed for efficient nutrition absorption. The objective of this study was to overexpress a β-mannanase gene directly in maize, the main ingredient of animal feed, to simplify the process of feed production.
The man5A gene encoding an excellent β-mannanase from acidophilic Bispora sp. MEY-1 was selected for heterologous overexpression. Expression of the modified gene (man5As) was driven by the embryo-specific promoter ZM-leg1A, and the transgene was transferred to three generations by backcrossing with commercial inbred Zheng58. Its exogenous integration into the maize embryonic genome and tissue specific expression in seeds were confirmed by PCR and Southern blot and Western blot analysis, respectively. Transgenic plants at BC3 generation showed agronomic traits statistically similar to Zheng58 except for less plant height (154.0 cm vs 158.3 cm). The expression level of MAN5AS reached up to 26,860 units per kilogram of maize seeds. Compared with its counterpart produced in Pichia pastoris, seed-derived MAN5AS had higher temperature optimum (90°C), and remained more β-mannanase activities after pelleting at 80°C, 100°C or 120°C.
This study shows the genetically stable overexpression of a fungal β-mannanase in maize and offers an effective and economic approach for transgene containment in maize for direct utilization without any purification or supplementation procedures.</description><subject>Agriculture</subject><subject>Agronomy</subject><subject>Animal Feed</subject><subject>Ascomycota - enzymology</subject><subject>Ascomycota - genetics</subject><subject>beta-Mannosidase - genetics</subject><subject>beta-Mannosidase - metabolism</subject><subject>Biology</subject><subject>Biotechnology</subject><subject>Cell walls</subject><subject>Containment</subject><subject>Corn</subject><subject>Embryos</subject><subject>Enzyme Activation</subject><subject>Enzyme Stability</subject><subject>Enzymes</subject><subject>Feed additives</subject><subject>Feeds</subject><subject>Food additives</subject><subject>Fungi</subject><subject>Gene Expression</subject><subject>Genetic Vectors - genetics</subject><subject>Genomes</subject><subject>Genomics</subject><subject>Inbreeding</subject><subject>Laboratories</subject><subject>Mannanases</subject><subject>Nutrition</subject><subject>Pelleting</subject><subject>Phenotype</subject><subject>Pichia - genetics</subject><subject>Pichia pastoris</subject><subject>Purification</subject><subject>Regeneration</subject><subject>Seeds</subject><subject>Seeds - genetics</subject><subject>Seeds - physiology</subject><subject>Tobacco</subject><subject>Transformation, Genetic</subject><subject>Transgenic plants</subject><subject>Yeast</subject><subject>Zea mays</subject><subject>Zea mays - genetics</subject><subject>Zea mays - physiology</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</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><sourceid>DOA</sourceid><recordid>eNptUk1v1DAQjRCIlsI_QGCJC5cs8WedC1KpClQq6gUOnKyJM9l6ldjBTiq6P4sfwm_Cy6ZVizh55HnvzbzRK4qXtFpRfkzfbcIcPfSrMXhcVZVUVKhHxSGtOSsVq_jje_VB8SylTQZxrdTT4oBxUdVU8MPCXV5jxJ9jxJRc8CR0BEg3-zX05PevcgDvwUNC0sUwkA8ujSECSeOKfDn7XlLiPBnAbZEkxDYR8C1Bv70ZkNgriGAnjG4LU5Z-XjzpoE_4YnmPim8fz76efi4vLj-dn55clFYyNZUaWqlVI6jitWyhAYRjSoXUEixaluvGtlpoVDW0VlFspM4OKVJGla0VPype73XHPiSzXCkZyjlV2XPFMuJ8j2gDbMwY3QDxxgRw5u9HiGsDcXK2R1Mz1PnaGrsWRKOVRqEazXVTMdtZLbPW-2Xa3AzYWvRThP6B6MOOd1dmHa4Nl6oWlGaBt4tADD9mTJMZXLLY9-AxzHlvpjXP_uVu1pt_oP93J_YoG0NKEbu7ZWhldsm5ZZldcsySnEx7dd_IHek2KvwPhtbCew</recordid><startdate>20130211</startdate><enddate>20130211</enddate><creator>Xu, Xiaolu</creator><creator>Zhang, Yuhong</creator><creator>Meng, Qingchang</creator><creator>Meng, Kun</creator><creator>Zhang, Wei</creator><creator>Zhou, Xiaojin</creator><creator>Luo, Huiying</creator><creator>Chen, Rumei</creator><creator>Yang, Peilong</creator><creator>Yao, Bin</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</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>7QG</scope><scope>7QL</scope><scope>7QO</scope><scope>7RV</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TG</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</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>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</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>KB.</scope><scope>KB0</scope><scope>KL.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PDBOC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20130211</creationdate><title>Overexpression of a fungal β-mannanase from Bispora sp. MEY-1 in maize seeds and enzyme characterization</title><author>Xu, Xiaolu ; Zhang, Yuhong ; Meng, Qingchang ; Meng, Kun ; Zhang, Wei ; Zhou, Xiaojin ; Luo, Huiying ; Chen, Rumei ; Yang, Peilong ; Yao, Bin</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c526t-8ad586b416395dabaea7114585acec2711bcd848e69adc61eb582031e1216c963</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>Agriculture</topic><topic>Agronomy</topic><topic>Animal Feed</topic><topic>Ascomycota - enzymology</topic><topic>Ascomycota - genetics</topic><topic>beta-Mannosidase - genetics</topic><topic>beta-Mannosidase - metabolism</topic><topic>Biology</topic><topic>Biotechnology</topic><topic>Cell walls</topic><topic>Containment</topic><topic>Corn</topic><topic>Embryos</topic><topic>Enzyme Activation</topic><topic>Enzyme Stability</topic><topic>Enzymes</topic><topic>Feed additives</topic><topic>Feeds</topic><topic>Food additives</topic><topic>Fungi</topic><topic>Gene Expression</topic><topic>Genetic Vectors - genetics</topic><topic>Genomes</topic><topic>Genomics</topic><topic>Inbreeding</topic><topic>Laboratories</topic><topic>Mannanases</topic><topic>Nutrition</topic><topic>Pelleting</topic><topic>Phenotype</topic><topic>Pichia - genetics</topic><topic>Pichia pastoris</topic><topic>Purification</topic><topic>Regeneration</topic><topic>Seeds</topic><topic>Seeds - genetics</topic><topic>Seeds - physiology</topic><topic>Tobacco</topic><topic>Transformation, Genetic</topic><topic>Transgenic plants</topic><topic>Yeast</topic><topic>Zea mays</topic><topic>Zea mays - genetics</topic><topic>Zea mays - physiology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Xu, Xiaolu</creatorcontrib><creatorcontrib>Zhang, Yuhong</creatorcontrib><creatorcontrib>Meng, Qingchang</creatorcontrib><creatorcontrib>Meng, Kun</creatorcontrib><creatorcontrib>Zhang, Wei</creatorcontrib><creatorcontrib>Zhou, Xiaojin</creatorcontrib><creatorcontrib>Luo, Huiying</creatorcontrib><creatorcontrib>Chen, Rumei</creatorcontrib><creatorcontrib>Yang, Peilong</creatorcontrib><creatorcontrib>Yao, Bin</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Nursing & Allied Health Database</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Meteorological & Geoastrophysical Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Agricultural Science Collection</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology 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>Materials Science & Engineering Collection</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>Advanced Technologies & Aerospace Collection</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Materials Science Collection</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>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Materials Science Database</collection><collection>Nursing & Allied Health Database (Alumni Edition)</collection><collection>Meteorological & Geoastrophysical Abstracts - 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Xu, Xiaolu</au><au>Zhang, Yuhong</au><au>Meng, Qingchang</au><au>Meng, Kun</au><au>Zhang, Wei</au><au>Zhou, Xiaojin</au><au>Luo, Huiying</au><au>Chen, Rumei</au><au>Yang, Peilong</au><au>Yao, Bin</au><au>Li, Yi</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Overexpression of a fungal β-mannanase from Bispora sp. MEY-1 in maize seeds and enzyme characterization</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2013-02-11</date><risdate>2013</risdate><volume>8</volume><issue>2</issue><spage>e56146</spage><epage>e56146</epage><pages>e56146-e56146</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Mannans and heteromannans are widespread in plants cell walls and are well-known as anti-nutritional factors in animal feed. To remove these factors, it is common practice to incorporate endo-β-mannanase into feed for efficient nutrition absorption. The objective of this study was to overexpress a β-mannanase gene directly in maize, the main ingredient of animal feed, to simplify the process of feed production.
The man5A gene encoding an excellent β-mannanase from acidophilic Bispora sp. MEY-1 was selected for heterologous overexpression. Expression of the modified gene (man5As) was driven by the embryo-specific promoter ZM-leg1A, and the transgene was transferred to three generations by backcrossing with commercial inbred Zheng58. Its exogenous integration into the maize embryonic genome and tissue specific expression in seeds were confirmed by PCR and Southern blot and Western blot analysis, respectively. Transgenic plants at BC3 generation showed agronomic traits statistically similar to Zheng58 except for less plant height (154.0 cm vs 158.3 cm). The expression level of MAN5AS reached up to 26,860 units per kilogram of maize seeds. Compared with its counterpart produced in Pichia pastoris, seed-derived MAN5AS had higher temperature optimum (90°C), and remained more β-mannanase activities after pelleting at 80°C, 100°C or 120°C.
This study shows the genetically stable overexpression of a fungal β-mannanase in maize and offers an effective and economic approach for transgene containment in maize for direct utilization without any purification or supplementation procedures.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>23409143</pmid><doi>10.1371/journal.pone.0056146</doi><oa>free_for_read</oa></addata></record> |
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subjects | Agriculture Agronomy Animal Feed Ascomycota - enzymology Ascomycota - genetics beta-Mannosidase - genetics beta-Mannosidase - metabolism Biology Biotechnology Cell walls Containment Corn Embryos Enzyme Activation Enzyme Stability Enzymes Feed additives Feeds Food additives Fungi Gene Expression Genetic Vectors - genetics Genomes Genomics Inbreeding Laboratories Mannanases Nutrition Pelleting Phenotype Pichia - genetics Pichia pastoris Purification Regeneration Seeds Seeds - genetics Seeds - physiology Tobacco Transformation, Genetic Transgenic plants Yeast Zea mays Zea mays - genetics Zea mays - physiology |
title | Overexpression of a fungal β-mannanase from Bispora sp. MEY-1 in maize seeds and enzyme characterization |
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