<?xml version="1.0" encoding="ISO-8859-1"?><article xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance">
<front>
<journal-meta>
<journal-id>0123-3475</journal-id>
<journal-title><![CDATA[Revista Colombiana de Biotecnología]]></journal-title>
<abbrev-journal-title><![CDATA[Rev. colomb. biotecnol]]></abbrev-journal-title>
<issn>0123-3475</issn>
<publisher>
<publisher-name><![CDATA[Instituto de Biotecnología, Universidad Nacional de Colombia]]></publisher-name>
</publisher>
</journal-meta>
<article-meta>
<article-id>S0123-34752020000200024</article-id>
<article-id pub-id-type="doi">10.15446/rev.colomb.biote.v22n2.79488</article-id>
<title-group>
<article-title xml:lang="en"><![CDATA[Modification and Expression of Beta-1,4-Endoglucanase encoding sequences of fungal origin in Escherichia coli BL21]]></article-title>
<article-title xml:lang="es"><![CDATA[Modificación y expresión de secuencias codificantes de Beta-1,4-Endoglucanasa de origen fúngico en Escherichia coli BL21]]></article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Galeano Vanegas]]></surname>
<given-names><![CDATA[Narmer Fernando]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Gutiérrez Calle]]></surname>
<given-names><![CDATA[Natalia]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Restrepo Franco]]></surname>
<given-names><![CDATA[Gloria María]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
</contrib-group>
<aff id="Af1">
<institution><![CDATA[,Universidad Católica de Manizales  ]]></institution>
<addr-line><![CDATA[ ]]></addr-line>
<country>Colombia</country>
</aff>
<aff id="Af2">
<institution><![CDATA[,Universidad Católica de Manizales  ]]></institution>
<addr-line><![CDATA[ ]]></addr-line>
<country>Colombia</country>
</aff>
<aff id="Af3">
<institution><![CDATA[,Universidad Católica de Manizales  ]]></institution>
<addr-line><![CDATA[ ]]></addr-line>
<country>Colombia</country>
</aff>
<pub-date pub-type="pub">
<day>00</day>
<month>12</month>
<year>2020</year>
</pub-date>
<pub-date pub-type="epub">
<day>00</day>
<month>12</month>
<year>2020</year>
</pub-date>
<volume>22</volume>
<numero>2</numero>
<fpage>24</fpage>
<lpage>34</lpage>
<copyright-statement/>
<copyright-year/>
<self-uri xlink:href="http://www.scielo.org.co/scielo.php?script=sci_arttext&amp;pid=S0123-34752020000200024&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://www.scielo.org.co/scielo.php?script=sci_abstract&amp;pid=S0123-34752020000200024&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://www.scielo.org.co/scielo.php?script=sci_pdf&amp;pid=S0123-34752020000200024&amp;lng=en&amp;nrm=iso"></self-uri><abstract abstract-type="short" xml:lang="en"><p><![CDATA[ABSTRACT Lignocellulose is the main and most abundant component of biomass. Annually, 200 million tons are generated in the world. Colombia has a high production of lignocellulosic residues that can be used in many industrial processes such as bioethanol produc-tion, promoting the bioeconomy. The objective of the present work was to express lignocellulolytic enzymes of eukaryotic origin in Escherichia coli BL21 (DE3). Initially, endoglucanase eukaryotic genes were selected and modified using bioinformatics methods for their production in E. coli BL21 (DE3) and saccharification of pure cellulose substrates. The gene selected for its modification and expression was  egl B from the fungus Aspergillus nidulans. Subsequently the enzyme integrity was tested by 3D modeling and molecular docking, as well as the conformation of its active site and its affinity for substrates of interest. Finally, cloning of the modified gene in plasmid pET151 TOPO was made and transformed in the strain E. coli BL21 (DE3) where several lignocellulose degradation tests were carried out using semiquantitative methods for the enzyme activity in carboxymethylcellulose. The presence of the three genes of interest within the plasmid pET151 TOPO and within the transformed cells of E. coli TOP10 and E. coli BL21 (DE3) was verified by colony PCRs performed. The presence of this gen was corroborated by sequencing. Expression of the modified endoglucanase enzyme was achieved in E. coli BL21 (DE3) expression cells, in soluble and functional form, demonstrated by the hydrolysis of the CMC substrate.]]></p></abstract>
<abstract abstract-type="short" xml:lang="es"><p><![CDATA[RESUMEN La lignocelulosa es el componente principal y más abundante de la biomasa. Anualmente se generan 200 millones de toneladas en el mundo. Colombia tiene una alta producción de residuos lignocelulósicos que pueden ser utilizados en muchos procesos industriales como la producción de bioetanol, promoviendo la bioeconomía. El objetivo del presente trabajo fue expresar enzimas lignocelulolíticas de origen eucariota en Escherichia coli BL21 (DE3). Inicialmente, los genes eucariotas de endoglucanasa se seleccionaron y modificaron mediante métodos bioinformáticos para su producción en E. coli BL21 (DE3) y la sacarificación de sustratos de celulosa. El gen seleccionado para su modificación y expresión fue eglB del hongo Aspergillus nidulans. Posteriormente se evaluó la integridad de la enzima mediante modelado 3D y acoplamiento molecular, así como la conformación de su sitio activo y su afinidad por sustratos de interés. Finalmente, se realizó la clonación del gen modificado en el plásmido pET151 TOPO y se transformó en la cepa E. coli BL21 (DE3) donde se realizaron varios ensayos de degradación de lignocelulosa utilizando métodos semicuantita-tivos para la actividad enzimática en carboximetilcelulosa. La presencia del gen de interés dentro del plásmido pET151 TOPO y dentro de las células transformadas de E. coli TOP10 y E. coli BL21 (DE3) se verificó mediante PCR de colonia. La presencia de este gene se corroboró por secuenciación eglB. La expresión de la enzima endoglucanasa modificada se logró en células de E. coli BL21 (DE3), en forma soluble y funcional, demostrada por la hidrólisis del sustrato de CMC.]]></p></abstract>
<kwd-group>
<kwd lng="en"><![CDATA[Cellulose]]></kwd>
<kwd lng="en"><![CDATA[Heterologous expression]]></kwd>
<kwd lng="en"><![CDATA[Cellulolytic enzymes]]></kwd>
<kwd lng="es"><![CDATA[Celulosa]]></kwd>
<kwd lng="es"><![CDATA[Expresión heteróloga]]></kwd>
<kwd lng="es"><![CDATA[Enzimas celulolíticas]]></kwd>
</kwd-group>
</article-meta>
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<name>
<surname><![CDATA[Zelena]]></surname>
<given-names><![CDATA[K.]]></given-names>
</name>
<name>
<surname><![CDATA[Eisele]]></surname>
<given-names><![CDATA[N.]]></given-names>
</name>
<name>
<surname><![CDATA[Berger]]></surname>
<given-names><![CDATA[R. G]]></given-names>
</name>
</person-group>
<article-title xml:lang=""><![CDATA[Escherichia coli as a production host for novel enzymes from basidiomycota]]></article-title>
<source><![CDATA[Biotechnology Advances]]></source>
<year>2014</year>
<volume>32</volume>
<numero>8</numero>
<issue>8</issue>
<page-range>1382-95</page-range></nlm-citation>
</ref>
</ref-list>
</back>
</article>
