Photoautotrophic production of macular pigment in a Chlamydomonas reinhardtii strain generated by using DNA‐free CRISPR‐Cas9 RNP‐mediated mutagenesis
Lutein and zeaxanthin are dietary carotenoids reported to be protective against age‐related macular degeneration. Recently, the green alga Chlamydomonas reinhardtii has received attention as a photosynthetic cell factory, but the potential of this alga for carotenoid production has not yet been eval...
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| Veröffentlicht in: | Biotechnology and bioengineering 2018-03, Vol.115 (3), p.719-728 |
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| creator | Baek, Kwangryul Yu, Jihyeon Jeong, Jooyeon Sim, Sang Jun Bae, Sangsu Jin, EonSeon |
| description | Lutein and zeaxanthin are dietary carotenoids reported to be protective against age‐related macular degeneration. Recently, the green alga Chlamydomonas reinhardtii has received attention as a photosynthetic cell factory, but the potential of this alga for carotenoid production has not yet been evaluated. In this study, we selected the C. reinhardtii CC‐4349 strain as the best candidate among seven laboratory strains tested for carotenoid production. A knock‐out mutant of the zeaxanthin epoxidase gene induced by preassembled DNA‐free CRISPR‐Cas9 ribonucleoproteins in the CC‐4349 strain had a significantly higher zeaxanthin content (56‐fold) and productivity (47‐fold) than the wild type without the reduction in lutein level. Furthermore, we produced eggs fortified with lutein (2‐fold) and zeaxanthin (2.2‐fold) by feeding hens a diet containing the mutant. Our results clearly demonstrate the possibility of cost‐effective commercial use of microalgal mutants induced by DNA‐free CRISPR‐Cas9 ribonucleoproteins in algal biotechnology for the production of high‐value products.
CC‐4349 is the best Chlamydomonas reinhardtii strain for carotenoid production. ZEP knockout mutants of CC‐4349 induced by DNA‐free CRISPR‐Cas9 RNP have a significantly higher zeaxanthin content and productivity than the wild‐type without the reduction in lutein level. Eggs are enriched with macular pigment by feeding hens the mutant‐containing diet. Approaches to circumvent GMO regulations in algal biotechnology are discussed. |
| doi_str_mv | 10.1002/bit.26499 |
| format | Article |
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CC‐4349 is the best Chlamydomonas reinhardtii strain for carotenoid production. ZEP knockout mutants of CC‐4349 induced by DNA‐free CRISPR‐Cas9 RNP have a significantly higher zeaxanthin content and productivity than the wild‐type without the reduction in lutein level. Eggs are enriched with macular pigment by feeding hens the mutant‐containing diet. Approaches to circumvent GMO regulations in algal biotechnology are discussed.</description><identifier>ISSN: 0006-3592</identifier><identifier>EISSN: 1097-0290</identifier><identifier>DOI: 10.1002/bit.26499</identifier><identifier>PMID: 29150930</identifier><language>eng</language><publisher>United States: Wiley Subscription Services, Inc</publisher><subject>Age ; Algae ; algal biotechnology ; Biotechnology ; Carotenoids ; Chlamydomonas reinhardtii ; Chlorophyll ; CRISPR ; CRISPR‐Cas9 ; Deoxyribonucleic acid ; Diet ; DNA ; Eggs ; Lutein ; Macular degeneration ; Mutagenesis ; Mutants ; Photosynthesis ; Ribonucleoproteins ; Zeaxanthin ; Zeaxanthin epoxidase</subject><ispartof>Biotechnology and bioengineering, 2018-03, Vol.115 (3), p.719-728</ispartof><rights>2017 Wiley Periodicals, Inc.</rights><rights>2018 Wiley Periodicals, Inc.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3909-4c67998bea418c9ac2ced05b1cbe491a67c672e55f1bc552a391d26b3da360a43</citedby><cites>FETCH-LOGICAL-c3909-4c67998bea418c9ac2ced05b1cbe491a67c672e55f1bc552a391d26b3da360a43</cites><orcidid>0000-0002-8366-0666 ; 0000-0001-5691-0124</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Fbit.26499$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fbit.26499$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>315,782,786,1419,27931,27932,45581,45582</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/29150930$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Baek, Kwangryul</creatorcontrib><creatorcontrib>Yu, Jihyeon</creatorcontrib><creatorcontrib>Jeong, Jooyeon</creatorcontrib><creatorcontrib>Sim, Sang Jun</creatorcontrib><creatorcontrib>Bae, Sangsu</creatorcontrib><creatorcontrib>Jin, EonSeon</creatorcontrib><title>Photoautotrophic production of macular pigment in a Chlamydomonas reinhardtii strain generated by using DNA‐free CRISPR‐Cas9 RNP‐mediated mutagenesis</title><title>Biotechnology and bioengineering</title><addtitle>Biotechnol Bioeng</addtitle><description>Lutein and zeaxanthin are dietary carotenoids reported to be protective against age‐related macular degeneration. Recently, the green alga Chlamydomonas reinhardtii has received attention as a photosynthetic cell factory, but the potential of this alga for carotenoid production has not yet been evaluated. In this study, we selected the C. reinhardtii CC‐4349 strain as the best candidate among seven laboratory strains tested for carotenoid production. A knock‐out mutant of the zeaxanthin epoxidase gene induced by preassembled DNA‐free CRISPR‐Cas9 ribonucleoproteins in the CC‐4349 strain had a significantly higher zeaxanthin content (56‐fold) and productivity (47‐fold) than the wild type without the reduction in lutein level. Furthermore, we produced eggs fortified with lutein (2‐fold) and zeaxanthin (2.2‐fold) by feeding hens a diet containing the mutant. Our results clearly demonstrate the possibility of cost‐effective commercial use of microalgal mutants induced by DNA‐free CRISPR‐Cas9 ribonucleoproteins in algal biotechnology for the production of high‐value products.
CC‐4349 is the best Chlamydomonas reinhardtii strain for carotenoid production. ZEP knockout mutants of CC‐4349 induced by DNA‐free CRISPR‐Cas9 RNP have a significantly higher zeaxanthin content and productivity than the wild‐type without the reduction in lutein level. Eggs are enriched with macular pigment by feeding hens the mutant‐containing diet. Approaches to circumvent GMO regulations in algal biotechnology are discussed.</description><subject>Age</subject><subject>Algae</subject><subject>algal biotechnology</subject><subject>Biotechnology</subject><subject>Carotenoids</subject><subject>Chlamydomonas reinhardtii</subject><subject>Chlorophyll</subject><subject>CRISPR</subject><subject>CRISPR‐Cas9</subject><subject>Deoxyribonucleic acid</subject><subject>Diet</subject><subject>DNA</subject><subject>Eggs</subject><subject>Lutein</subject><subject>Macular degeneration</subject><subject>Mutagenesis</subject><subject>Mutants</subject><subject>Photosynthesis</subject><subject>Ribonucleoproteins</subject><subject>Zeaxanthin</subject><subject>Zeaxanthin epoxidase</subject><issn>0006-3592</issn><issn>1097-0290</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2018</creationdate><recordtype>article</recordtype><recordid>eNp1kc9OGzEQxi1UVELogReoLPXUw4Lt_Zc5hm2BSAiiEM6rWa83cZRdB9urKjcegXvfrk-C86e99TQz0m--0TcfIZecXXHGxHWl_ZXIEoATMuAM8ogJYJ_IgDGWRXEK4oycO7cKYz7Kss_kTABPGcRsQH5Pl8Yb7L3x1myWWtKNNXUvvTYdNQ1tUfZrtHSjF63qPNUdRVos19hua9OaDh21SndLtLXXmjpvMSAL1SmLXtW02tLe6W5BfzyO_7y9N1YpWswmz9NZmAp0QGeP09C2qtb7hbb3uFt32l2Q0wbXTn051iF5uf05L-6jh6e7STF-iGQMDKJEZjnAqFKY8JEElEKqmqUVl5VKgGOWB0CoNG14JdNUYAy8FlkV1xhnDJN4SL4ddIP11145X65Mb7twsuQAgvOY5zvq-4GS1jhnVVNurG7RbkvOyl0MZYih3McQ2K9Hxb4Kzv6Rf_8egOsD8Euv1fb_SuXNZH6Q_ACPgJbj</recordid><startdate>201803</startdate><enddate>201803</enddate><creator>Baek, Kwangryul</creator><creator>Yu, Jihyeon</creator><creator>Jeong, Jooyeon</creator><creator>Sim, Sang Jun</creator><creator>Bae, Sangsu</creator><creator>Jin, EonSeon</creator><general>Wiley Subscription Services, Inc</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QF</scope><scope>7QO</scope><scope>7QQ</scope><scope>7SC</scope><scope>7SE</scope><scope>7SP</scope><scope>7SR</scope><scope>7T7</scope><scope>7TA</scope><scope>7TB</scope><scope>7U5</scope><scope>8BQ</scope><scope>8FD</scope><scope>C1K</scope><scope>F28</scope><scope>FR3</scope><scope>H8D</scope><scope>H8G</scope><scope>JG9</scope><scope>JQ2</scope><scope>KR7</scope><scope>L7M</scope><scope>L~C</scope><scope>L~D</scope><scope>P64</scope><orcidid>https://orcid.org/0000-0002-8366-0666</orcidid><orcidid>https://orcid.org/0000-0001-5691-0124</orcidid></search><sort><creationdate>201803</creationdate><title>Photoautotrophic production of macular pigment in a Chlamydomonas reinhardtii strain generated by using DNA‐free CRISPR‐Cas9 RNP‐mediated mutagenesis</title><author>Baek, Kwangryul ; Yu, Jihyeon ; Jeong, Jooyeon ; Sim, Sang Jun ; Bae, Sangsu ; Jin, EonSeon</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3909-4c67998bea418c9ac2ced05b1cbe491a67c672e55f1bc552a391d26b3da360a43</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2018</creationdate><topic>Age</topic><topic>Algae</topic><topic>algal biotechnology</topic><topic>Biotechnology</topic><topic>Carotenoids</topic><topic>Chlamydomonas reinhardtii</topic><topic>Chlorophyll</topic><topic>CRISPR</topic><topic>CRISPR‐Cas9</topic><topic>Deoxyribonucleic acid</topic><topic>Diet</topic><topic>DNA</topic><topic>Eggs</topic><topic>Lutein</topic><topic>Macular degeneration</topic><topic>Mutagenesis</topic><topic>Mutants</topic><topic>Photosynthesis</topic><topic>Ribonucleoproteins</topic><topic>Zeaxanthin</topic><topic>Zeaxanthin epoxidase</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Baek, Kwangryul</creatorcontrib><creatorcontrib>Yu, Jihyeon</creatorcontrib><creatorcontrib>Jeong, Jooyeon</creatorcontrib><creatorcontrib>Sim, Sang Jun</creatorcontrib><creatorcontrib>Bae, Sangsu</creatorcontrib><creatorcontrib>Jin, EonSeon</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>Aluminium Industry Abstracts</collection><collection>Biotechnology Research Abstracts</collection><collection>Ceramic Abstracts</collection><collection>Computer and Information Systems Abstracts</collection><collection>Corrosion Abstracts</collection><collection>Electronics & Communications Abstracts</collection><collection>Engineered Materials Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Materials Business File</collection><collection>Mechanical & Transportation Engineering Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ANTE: Abstracts in New Technology & Engineering</collection><collection>Engineering Research Database</collection><collection>Aerospace Database</collection><collection>Copper Technical Reference Library</collection><collection>Materials Research Database</collection><collection>ProQuest Computer Science Collection</collection><collection>Civil Engineering Abstracts</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>Computer and Information Systems Abstracts Academic</collection><collection>Computer and Information Systems Abstracts Professional</collection><collection>Biotechnology and BioEngineering Abstracts</collection><jtitle>Biotechnology and bioengineering</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Baek, Kwangryul</au><au>Yu, Jihyeon</au><au>Jeong, Jooyeon</au><au>Sim, Sang Jun</au><au>Bae, Sangsu</au><au>Jin, EonSeon</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Photoautotrophic production of macular pigment in a Chlamydomonas reinhardtii strain generated by using DNA‐free CRISPR‐Cas9 RNP‐mediated mutagenesis</atitle><jtitle>Biotechnology and bioengineering</jtitle><addtitle>Biotechnol Bioeng</addtitle><date>2018-03</date><risdate>2018</risdate><volume>115</volume><issue>3</issue><spage>719</spage><epage>728</epage><pages>719-728</pages><issn>0006-3592</issn><eissn>1097-0290</eissn><abstract>Lutein and zeaxanthin are dietary carotenoids reported to be protective against age‐related macular degeneration. Recently, the green alga Chlamydomonas reinhardtii has received attention as a photosynthetic cell factory, but the potential of this alga for carotenoid production has not yet been evaluated. In this study, we selected the C. reinhardtii CC‐4349 strain as the best candidate among seven laboratory strains tested for carotenoid production. A knock‐out mutant of the zeaxanthin epoxidase gene induced by preassembled DNA‐free CRISPR‐Cas9 ribonucleoproteins in the CC‐4349 strain had a significantly higher zeaxanthin content (56‐fold) and productivity (47‐fold) than the wild type without the reduction in lutein level. Furthermore, we produced eggs fortified with lutein (2‐fold) and zeaxanthin (2.2‐fold) by feeding hens a diet containing the mutant. Our results clearly demonstrate the possibility of cost‐effective commercial use of microalgal mutants induced by DNA‐free CRISPR‐Cas9 ribonucleoproteins in algal biotechnology for the production of high‐value products.
CC‐4349 is the best Chlamydomonas reinhardtii strain for carotenoid production. ZEP knockout mutants of CC‐4349 induced by DNA‐free CRISPR‐Cas9 RNP have a significantly higher zeaxanthin content and productivity than the wild‐type without the reduction in lutein level. Eggs are enriched with macular pigment by feeding hens the mutant‐containing diet. Approaches to circumvent GMO regulations in algal biotechnology are discussed.</abstract><cop>United States</cop><pub>Wiley Subscription Services, Inc</pub><pmid>29150930</pmid><doi>10.1002/bit.26499</doi><tpages>10</tpages><orcidid>https://orcid.org/0000-0002-8366-0666</orcidid><orcidid>https://orcid.org/0000-0001-5691-0124</orcidid></addata></record> |
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| ispartof | Biotechnology and bioengineering, 2018-03, Vol.115 (3), p.719-728 |
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| subjects | Age Algae algal biotechnology Biotechnology Carotenoids Chlamydomonas reinhardtii Chlorophyll CRISPR CRISPR‐Cas9 Deoxyribonucleic acid Diet DNA Eggs Lutein Macular degeneration Mutagenesis Mutants Photosynthesis Ribonucleoproteins Zeaxanthin Zeaxanthin epoxidase |
| title | Photoautotrophic production of macular pigment in a Chlamydomonas reinhardtii strain generated by using DNA‐free CRISPR‐Cas9 RNP‐mediated mutagenesis |
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