<?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>0122-7483</journal-id>
<journal-title><![CDATA[Universitas Scientiarum]]></journal-title>
<abbrev-journal-title><![CDATA[Univ. Sci.]]></abbrev-journal-title>
<issn>0122-7483</issn>
<publisher>
<publisher-name><![CDATA[Facultad de Ciencias de la Pontificia Universidad Javeriana de Bogotá.]]></publisher-name>
</publisher>
</journal-meta>
<article-meta>
<article-id>S0122-74832016000300195</article-id>
<article-id pub-id-type="doi">10.11144/Javeriana.SC21-3.corh</article-id>
<title-group>
<article-title xml:lang="en"><![CDATA[Characterization of recombinant human lysosomal beta-hexosaminidases produced in the methylotrophic yeast Pichia pastoris]]></article-title>
<article-title xml:lang="es"><![CDATA[Caracterización de beta-hexosaminidasas lisosomales humanas recombinantes producidas en la levadura metilotrófica Pichia pastoris]]></article-title>
<article-title xml:lang="pt"><![CDATA[Caracterização de beta-hexosaminidases lisossomais humanas recombinantes produzidas na levedura metilotrófica Pichia pastoris]]></article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Espejo-Mojica]]></surname>
<given-names><![CDATA[Angela J.]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Mosquera]]></surname>
<given-names><![CDATA[Angela]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Rodrfguez-Lopez]]></surname>
<given-names><![CDATA[Alexander]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
<xref ref-type="aff" rid="Aaf"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Diaz]]></surname>
<given-names><![CDATA[Dennis]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Beltran]]></surname>
<given-names><![CDATA[Laura]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Hernandez]]></surname>
<given-names><![CDATA[Francy Liliana]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Alméciga-Diaz]]></surname>
<given-names><![CDATA[Carlos J.]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Barrera]]></surname>
<given-names><![CDATA[Luis A.]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
<xref ref-type="aff" rid="Aaf"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Salcedo-Reyes]]></surname>
<given-names><![CDATA[Juan Carlos]]></given-names>
</name>
</contrib>
</contrib-group>
<aff id="Af1">
<institution><![CDATA[,Pontificia Universidad Javeriana  ]]></institution>
<addr-line><![CDATA[Bogotá ]]></addr-line>
<country>Colombia</country>
</aff>
<aff id="Af2">
<institution><![CDATA[,Pontificia Universidad Javeriana  ]]></institution>
<addr-line><![CDATA[Bogotá ]]></addr-line>
<country>Colombia</country>
</aff>
<aff id="Af3">
<institution><![CDATA[,Hospital Universitario San Ignacio  ]]></institution>
<addr-line><![CDATA[Bogotá ]]></addr-line>
<country>Colombia</country>
</aff>
<pub-date pub-type="pub">
<day>00</day>
<month>12</month>
<year>2016</year>
</pub-date>
<pub-date pub-type="epub">
<day>00</day>
<month>12</month>
<year>2016</year>
</pub-date>
<volume>21</volume>
<numero>3</numero>
<fpage>195</fpage>
<lpage>217</lpage>
<copyright-statement/>
<copyright-year/>
<self-uri xlink:href="http://www.scielo.org.co/scielo.php?script=sci_arttext&amp;pid=S0122-74832016000300195&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://www.scielo.org.co/scielo.php?script=sci_abstract&amp;pid=S0122-74832016000300195&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://www.scielo.org.co/scielo.php?script=sci_pdf&amp;pid=S0122-74832016000300195&amp;lng=en&amp;nrm=iso"></self-uri><abstract abstract-type="short" xml:lang="en"><p><![CDATA[Abstract &#946;-hexosaminidases (Hex) are dimeric enzymes involved in the lysosomal degradation of glycolipids and glycans. They are formed by &#945;- and/or &#946;-subunits encoded by HEXA and HEXB genes, respectively. Mutations in these genes lead to Tay Sachs or Sandhoff diseases, which are neurodegenerative disorders caused by the accumulation of non-degraded glycolipids. Although tissue-derived Hex have been widely characterized, limited information is available for recombinant &#945;-hexosaminidases. In this study, human lysosomal recombinant Hex (rhHex-A, rhHex-B, and rhHex-S) were produced in the methylotrophic yeast Pichia pastoris GS115. The highest specific enzyme activities were 13,124 for rhHexA; 12,779 for rhHex-B; and 14,606 U .mg-1 for rhHex-S. These results were 25- to 50-fold higher than those obtained from normal human leukocytes. Proteins were purified and characterized at different pH and temperature conditions. All proteins were stable at acidic pH, and at 4 °C and 37 °C. At 45 °C rhHex-S was completely inactivated, while rhHex-A and rhHex-B showed high stability. This study demonstrates P. pastoris GS115 potential for polymeric lysosomal enzyme production, and describes the characterization of recombinant &#946;-hexosaminidases produced within the same host.]]></p></abstract>
<abstract abstract-type="short" xml:lang="es"><p><![CDATA[Resumen Las &#946;-hexosaminidasas (Hex) son enzimas diméricas involucradas en la degradación lisosomal de glicolípidos y glicanos. Estas enzimas están formadas por las subunidades &#945;- y/o &#946;-codificadas por los genes HEXA and HEXB respectivamente. Las mutaciones de estos genes conducen a las enfermedades de Tay Sachs o Sandhoff, que son desórdenes neurodegenerativos causados por la acumulación de glicolípidos no degradados. Aunque las Hex derivadas de tejido han sido ampliamente caracterizadas, la información disponible sobre las p-hexosaminidasas recombinantes es limitada. En este estudio se produjeron Hex recombinantes lisosomales (rhHex-A, rhHex-B y rhHex-S) en la levadura metilotrófica Pichia pastoris GS115. Las actividades específicas más altas de las enzimas fueron 13.124, 12.779, 14.606 U .mg-1 para rhHex-A, rhHex-B y rhHex-S, respectivamente. Estos resultados fueron 25 a 50 veces más altos que los obtenidos de leucocitos humanos normales. Las proteínas se purificaron y se caracterizaron a diferentes condiciones de pH y temperatura. Todas las proteínas fueron estables a pH ácido y a 4°C y 37°C. A 45°C la rhHex-S se inactivó completamente, mientras que rhHex-A y rhHex-B mostraron alta estabilidad. Este estudio demuestra el potencial de P. pastoris GS115 para la producción de enzimas lisosomales poliméricas y presenta la caracterización de distintas &#946;-hexosaminidasas recombinantes producidas en un único hospedero.]]></p></abstract>
<abstract abstract-type="short" xml:lang="pt"><p><![CDATA[Resumen As &#946;-hexosaminidases (Hex) são enzimas diméricas envolvidas na degradação lisossomal de glicolipídeos e glicanos. Essas enzimas são formadas por subunidades a- e/ou p-codificadas pelos genes HEXA e HEXB, respectivamente. As mutações nesses genes causam a doença de Sandhoff ou Tay Sachs, que são desordens neurodegenerativas causadas pela acumulação de glicolipídeos não degradados. Embora Hex derivadas de tecido hajam sido caracterizadas extensivamente, as informações disponíveis sobre as p-hexosaminidases recombinantes são limitadas. Esse estudo produziu Hex recombinantes lisossomais (rhHex-A, rhHex-B e rhHex-S) na levedura metilotrófica Pichia pastoris GS115. As atividades específicas mais altas das enzimas foram 13.124, 12.779, 14.606 U .mg-1 para rhHex-A, rhHex-B y rhHex-S, respectivamente. Esses resultados foram 25 a 50 vezes mais altos do que os obtidos a partir de leucócitos humanos normais. As proteínas foram purificadas e caracterizadas em diferentes condições de pH e temperatura. Todas as proteínas foram estáveis a pH ácido e a 4°C e 37°C. A 45°C a rhHex-S foi completamente inativada, enquanto rhHex rhHex-A e B se mostraram altamente estáveis. Esse estudo demonstra o potencial de P. pastoris GS115 para a produção de enzimas lisossomais poliméricas e apresenta a caracterização de diferentes p-hexosaminidases recombinantes produzidas em único hospedeiro.]]></p></abstract>
<kwd-group>
<kwd lng="en"><![CDATA[&#946;-N-acetylhexosaminidases]]></kwd>
<kwd lng="en"><![CDATA[characterization]]></kwd>
<kwd lng="en"><![CDATA[Pichia pastoris, recombinant hexosaminidases]]></kwd>
<kwd lng="en"><![CDATA[Sandhoff disease]]></kwd>
<kwd lng="en"><![CDATA[Tay Sachs disease.]]></kwd>
<kwd lng="es"><![CDATA[&#946;-N-acetylhexosaminidasas]]></kwd>
<kwd lng="es"><![CDATA[caracterización]]></kwd>
<kwd lng="es"><![CDATA[Pichia pastoris-, hexosaminidasas recombinantes]]></kwd>
<kwd lng="es"><![CDATA[enfermedad de Sandhoff]]></kwd>
<kwd lng="es"><![CDATA[enfermedad de Tay Sachs.]]></kwd>
<kwd lng="pt"><![CDATA[&#946;-N-acetilhexosaminidases]]></kwd>
<kwd lng="pt"><![CDATA[caracterização]]></kwd>
<kwd lng="pt"><![CDATA[Pichia pastoris]]></kwd>
<kwd lng="pt"><![CDATA[hexosaminidases recombinantes]]></kwd>
<kwd lng="pt"><![CDATA[Doença de Sandhoff]]></kwd>
<kwd lng="pt"><![CDATA[Doenca de Tay Sachs.]]></kwd>
</kwd-group>
</article-meta>
</front><back>
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