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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Altered lipid composition and enhanced lipid production in green microalga by introduction of brassica diacylglycerol acyltransferase 2</title>
<author><name sortKey="Ahmad, Irshad" sort="Ahmad, Irshad" uniqKey="Ahmad I" first="Irshad" last="Ahmad">Irshad Ahmad</name>
<affiliation><nlm:aff id="A1">International Centre for Genetic Engineering and Biotechnology, New Delhi, India</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Sharma, Anil K" sort="Sharma, Anil K" uniqKey="Sharma A" first="Anil K." last="Sharma">Anil K. Sharma</name>
<affiliation><nlm:aff id="A2">Department of Biotechnology, M.M. University, Mullana, Haryana, India</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Daniell, Henry" sort="Daniell, Henry" uniqKey="Daniell H" first="Henry" last="Daniell">Henry Daniell</name>
<affiliation><nlm:aff id="A3">Department of Biochemistry and Pathology, School of Dental Medicine, University of Pennsylvania, Philadelphia, PA, USA</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Kumar, Shashi" sort="Kumar, Shashi" uniqKey="Kumar S" first="Shashi" last="Kumar">Shashi Kumar</name>
<affiliation><nlm:aff id="A1">International Centre for Genetic Engineering and Biotechnology, New Delhi, India</nlm:aff>
</affiliation>
</author>
</titleStmt>
<publicationStmt><idno type="wicri:source">PMC</idno>
<idno type="pmid">25403771</idno>
<idno type="pmc">4599878</idno>
<idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4599878</idno>
<idno type="RBID">PMC:4599878</idno>
<idno type="doi">10.1111/pbi.12278</idno>
<date when="2014">2014</date>
<idno type="wicri:Area/Pmc/Corpus">000D63</idno>
</publicationStmt>
<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Altered lipid composition and enhanced lipid production in green microalga by introduction of brassica diacylglycerol acyltransferase 2</title>
<author><name sortKey="Ahmad, Irshad" sort="Ahmad, Irshad" uniqKey="Ahmad I" first="Irshad" last="Ahmad">Irshad Ahmad</name>
<affiliation><nlm:aff id="A1">International Centre for Genetic Engineering and Biotechnology, New Delhi, India</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Sharma, Anil K" sort="Sharma, Anil K" uniqKey="Sharma A" first="Anil K." last="Sharma">Anil K. Sharma</name>
<affiliation><nlm:aff id="A2">Department of Biotechnology, M.M. University, Mullana, Haryana, India</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Daniell, Henry" sort="Daniell, Henry" uniqKey="Daniell H" first="Henry" last="Daniell">Henry Daniell</name>
<affiliation><nlm:aff id="A3">Department of Biochemistry and Pathology, School of Dental Medicine, University of Pennsylvania, Philadelphia, PA, USA</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Kumar, Shashi" sort="Kumar, Shashi" uniqKey="Kumar S" first="Shashi" last="Kumar">Shashi Kumar</name>
<affiliation><nlm:aff id="A1">International Centre for Genetic Engineering and Biotechnology, New Delhi, India</nlm:aff>
</affiliation>
</author>
</analytic>
<series><title level="j">Plant biotechnology journal</title>
<idno type="ISSN">1467-7644</idno>
<idno type="eISSN">1467-7652</idno>
<imprint><date when="2014">2014</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
</fileDesc>
<profileDesc><textClass></textClass>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en"><title>Summary</title>
<p id="P1">Higher lipid biosynthesis and accumulation are important to achieve economic viability of biofuel production <italic>via</italic>
microalgae. To enhance lipid content, <italic>Chlamydomonas reinhardtii</italic>
was genetically engineered with a key enzyme diacylglycerol acyltransferase (<italic>BnDGAT2</italic>
) from <italic>Brassica napus,</italic>
responsible for neutral lipid biosynthesis. The transformed colonies harbouring <italic>aph7</italic>
gene, screened on hygromycin-supplemented medium, achieved transformation frequency of ~120 ± 10 colonies/1 × 10<sup>6</sup>
cells. Transgene integration and expression were confirmed by PCR, Southern blots, staining lipid droplets, proteins and spectro-fluorometric analysis of Nile red-stained cells. The neutral lipid is a major class (over 80% of total lipids) and most significant requirement for biodiesel production; this was remarkably higher in the transformed alga than the untransformed control. The levels of saturated fatty acids in the transformed alga decreased to about 7% while unsaturated fatty acids increased proportionately when compared to wild type cells. Polyunsaturated fatty acids, especially α-linolenic acid, an essential omega-3 fatty acid, were enhanced up to 12% in the transformed line. Nile red staining confirmed formation of a large number of lipid globules in the transformed alga. Evaluation of long-term stability and vitality of the transgenic alga revealed that cryopreservation produced significantly higher quantity of lipid than those maintained continuously over 128 generations on solid medium. The overexpression of <italic>BnDGAT2</italic>
significantly altered the fatty acids profile in the transformed alga. Results of this study offer a valuable strategy of genetic manipulation for enhancing polyunsaturated fatty acids and neutral lipids for biofuel production in algae.</p>
</div>
</front>
</TEI>
<pmc article-type="research-article"><pmc-comment>The publisher of this article does not allow downloading of the full text in XML form.</pmc-comment>
<pmc-dir>properties manuscript</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-journal-id">101201889</journal-id>
<journal-id journal-id-type="pubmed-jr-id">31826</journal-id>
<journal-id journal-id-type="nlm-ta">Plant Biotechnol J</journal-id>
<journal-id journal-id-type="iso-abbrev">Plant Biotechnol. J.</journal-id>
<journal-title-group><journal-title>Plant biotechnology journal</journal-title>
</journal-title-group>
<issn pub-type="ppub">1467-7644</issn>
<issn pub-type="epub">1467-7652</issn>
</journal-meta>
<article-meta><article-id pub-id-type="pmid">25403771</article-id>
<article-id pub-id-type="pmc">4599878</article-id>
<article-id pub-id-type="doi">10.1111/pbi.12278</article-id>
<article-id pub-id-type="manuscript">NIHMS685816</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Article</subject>
</subj-group>
</article-categories>
<title-group><article-title>Altered lipid composition and enhanced lipid production in green microalga by introduction of brassica diacylglycerol acyltransferase 2</article-title>
</title-group>
<contrib-group><contrib contrib-type="author"><name><surname>Ahmad</surname>
<given-names>Irshad</given-names>
</name>
<xref ref-type="aff" rid="A1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Sharma</surname>
<given-names>Anil K.</given-names>
</name>
<xref ref-type="aff" rid="A2">2</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Daniell</surname>
<given-names>Henry</given-names>
</name>
<xref ref-type="aff" rid="A3">3</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Kumar</surname>
<given-names>Shashi</given-names>
</name>
<xref ref-type="aff" rid="A1">1</xref>
<xref ref-type="corresp" rid="cor1">*</xref>
</contrib>
</contrib-group>
<aff id="A1"><label>1</label>
International Centre for Genetic Engineering and Biotechnology, New Delhi, India</aff>
<aff id="A2"><label>2</label>
Department of Biotechnology, M.M. University, Mullana, Haryana, India</aff>
<aff id="A3"><label>3</label>
Department of Biochemistry and Pathology, School of Dental Medicine, University of Pennsylvania, Philadelphia, PA, USA</aff>
<author-notes><corresp id="cor1"><label>*</label>
<italic>Correspondence</italic>
. <email>skrhode@icgeb.res.in</email>
</corresp>
</author-notes>
<pub-date pub-type="nihms-submitted"><day>9</day>
<month>9</month>
<year>2015</year>
</pub-date>
<pub-date pub-type="epub"><day>18</day>
<month>11</month>
<year>2014</year>
</pub-date>
<pub-date pub-type="ppub"><month>5</month>
<year>2015</year>
</pub-date>
<pub-date pub-type="pmc-release"><day>01</day>
<month>11</month>
<year>2015</year>
</pub-date>
<volume>13</volume>
<issue>4</issue>
<fpage>540</fpage>
<lpage>550</lpage>
<pmc-comment>elocation-id from pubmed: 10.1111/pbi.12278</pmc-comment>
<abstract><title>Summary</title>
<p id="P1">Higher lipid biosynthesis and accumulation are important to achieve economic viability of biofuel production <italic>via</italic>
microalgae. To enhance lipid content, <italic>Chlamydomonas reinhardtii</italic>
was genetically engineered with a key enzyme diacylglycerol acyltransferase (<italic>BnDGAT2</italic>
) from <italic>Brassica napus,</italic>
responsible for neutral lipid biosynthesis. The transformed colonies harbouring <italic>aph7</italic>
gene, screened on hygromycin-supplemented medium, achieved transformation frequency of ~120 ± 10 colonies/1 × 10<sup>6</sup>
cells. Transgene integration and expression were confirmed by PCR, Southern blots, staining lipid droplets, proteins and spectro-fluorometric analysis of Nile red-stained cells. The neutral lipid is a major class (over 80% of total lipids) and most significant requirement for biodiesel production; this was remarkably higher in the transformed alga than the untransformed control. The levels of saturated fatty acids in the transformed alga decreased to about 7% while unsaturated fatty acids increased proportionately when compared to wild type cells. Polyunsaturated fatty acids, especially α-linolenic acid, an essential omega-3 fatty acid, were enhanced up to 12% in the transformed line. Nile red staining confirmed formation of a large number of lipid globules in the transformed alga. Evaluation of long-term stability and vitality of the transgenic alga revealed that cryopreservation produced significantly higher quantity of lipid than those maintained continuously over 128 generations on solid medium. The overexpression of <italic>BnDGAT2</italic>
significantly altered the fatty acids profile in the transformed alga. Results of this study offer a valuable strategy of genetic manipulation for enhancing polyunsaturated fatty acids and neutral lipids for biofuel production in algae.</p>
</abstract>
<kwd-group><kwd>green algae</kwd>
<kwd>bioenergy</kwd>
<kwd>biodiesel</kwd>
<kwd>transgenic algae</kwd>
<kwd>polyunsaturated fatty acids</kwd>
<kwd>neutral lipids</kwd>
</kwd-group>
</article-meta>
</front>
</pmc>
</record>
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