Expression of a Recombinant Human Glycosyltransferase from a Synthetic Gene and its Utilization for Synthesis of the Human Blood Group B Trisaccharide
A 1034‐bp synthetic gene encoding the human blood group B glycosyltransferase, which catalyzes the transfer of galactose from UDP‐Gal to Fucα(l‐2)Galβ‐OR to give the blood group B determinant Galα(l‐3)[Fucα(1‐2)]Gal β‐OR (where R is a glycoprotein or glycolipid), has been expressed in Escherichia co...
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| Veröffentlicht in: | European journal of biochemistry 1995-11, Vol.234 (1), p.323-328 |
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| container_end_page | 328 |
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| container_title | European journal of biochemistry |
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| creator | Seto, Nina O. L. Palcic, Monica M. Hindsgaul, Ole Bundle, David R. Narang, Saran A. |
| description | A 1034‐bp synthetic gene encoding the human blood group B glycosyltransferase, which catalyzes the transfer of galactose from UDP‐Gal to Fucα(l‐2)Galβ‐OR to give the blood group B determinant Galα(l‐3)[Fucα(1‐2)]Gal β‐OR (where R is a glycoprotein or glycolipid), has been expressed in Escherichia coli by replacing its membrane‐anchoring domain with an omp A bacterial secretory signal. The active enzyme was purified from the periplasm using UDP‐hexanolamine affinity chromatography and used in the synthesis of preparative amounts of the human blood group B trisaccharide antigen. The substrate specificity and kinetics of the recombinant enzyme were comparable to the enzyme from human sera. Thus we have achieved the construction of a completely synthetic glycosyltransferase gene and its successful expression. |
| doi_str_mv | 10.1111/j.1432-1033.1995.323_c.x |
| format | Article |
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L.</creatorcontrib><creatorcontrib>Palcic, Monica M.</creatorcontrib><creatorcontrib>Hindsgaul, Ole</creatorcontrib><creatorcontrib>Bundle, David R.</creatorcontrib><creatorcontrib>Narang, Saran A.</creatorcontrib><title>Expression of a Recombinant Human Glycosyltransferase from a Synthetic Gene and its Utilization for Synthesis of the Human Blood Group B Trisaccharide</title><title>European journal of biochemistry</title><addtitle>Eur J Biochem</addtitle><description>A 1034‐bp synthetic gene encoding the human blood group B glycosyltransferase, which catalyzes the transfer of galactose from UDP‐Gal to Fucα(l‐2)Galβ‐OR to give the blood group B determinant Galα(l‐3)[Fucα(1‐2)]Gal β‐OR (where R is a glycoprotein or glycolipid), has been expressed in Escherichia coli by replacing its membrane‐anchoring domain with an omp A bacterial secretory signal. The active enzyme was purified from the periplasm using UDP‐hexanolamine affinity chromatography and used in the synthesis of preparative amounts of the human blood group B trisaccharide antigen. The substrate specificity and kinetics of the recombinant enzyme were comparable to the enzyme from human sera. Thus we have achieved the construction of a completely synthetic glycosyltransferase gene and its successful expression.</description><subject>ABO Blood-Group System</subject><subject>Amino Acid Sequence</subject><subject>Base Sequence</subject><subject>blood group antigen</subject><subject>Carbohydrate Sequence</subject><subject>Cloning, Molecular</subject><subject>DNA, Recombinant</subject><subject>enzymic synthesis</subject><subject>Escherichia coli - genetics</subject><subject>Galactosyltransferases - genetics</subject><subject>Genes, Synthetic</subject><subject>glycosyltransferases</subject><subject>Humans</subject><subject>Kinetics</subject><subject>Molecular Sequence Data</subject><subject>recombinant</subject><subject>Recombinant Proteins - genetics</subject><subject>Recombinant Proteins - metabolism</subject><subject>synthetic genes</subject><subject>Trisaccharides - biosynthesis</subject><issn>0014-2956</issn><issn>1432-1033</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1995</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNUctq4zAUFUNLJ23nEwa06s6uZFlWvBqakKaFQqGPtZDlK6pgSxnJpvF8yHxv7cZ03bu5B87jwj0IYUpSOs71LqU5yxJKGEtpWfKUZUzq9PADLb6IE7QghOZJVvLiJzqPcUcIKcpCnKGzJc_KoiAL9H9z2AeI0XqHvcEKP4H2bWWdch2-61vl8LYZtI9D0wXlooGgImATfDuKnwfXvUFnNd6CA6xcjW0X8WtnG_tPdVOo8WGWRRunEyOag1eN9zXeBt_v8Qq_BBuV1m8q2Bou0alRTYRf875Ar7ebl_Vd8vC4vV_fPCQ659k64cyYSvHaCK6ZKQXjOSc5rxQTFehag8hLTgpSGF1mRlBOac1MXfAlYRUVwC7Q1TF3H_zfHmInWxs1NI1y4PsohRA5E4KNwuVRqIOPMYCR-2BbFQZJiZwqkTs5fV5On5dTJfKzEnkYrb_nG33VQv1lnDsY-T9H_t02MHw7V95uVs8TXrMP77GeYQ</recordid><startdate>19951115</startdate><enddate>19951115</enddate><creator>Seto, Nina O. 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L.</creatorcontrib><creatorcontrib>Palcic, Monica M.</creatorcontrib><creatorcontrib>Hindsgaul, Ole</creatorcontrib><creatorcontrib>Bundle, David R.</creatorcontrib><creatorcontrib>Narang, Saran A.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>European journal of biochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Seto, Nina O. L.</au><au>Palcic, Monica M.</au><au>Hindsgaul, Ole</au><au>Bundle, David R.</au><au>Narang, Saran A.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Expression of a Recombinant Human Glycosyltransferase from a Synthetic Gene and its Utilization for Synthesis of the Human Blood Group B Trisaccharide</atitle><jtitle>European journal of biochemistry</jtitle><addtitle>Eur J Biochem</addtitle><date>1995-11-15</date><risdate>1995</risdate><volume>234</volume><issue>1</issue><spage>323</spage><epage>328</epage><pages>323-328</pages><issn>0014-2956</issn><eissn>1432-1033</eissn><abstract>A 1034‐bp synthetic gene encoding the human blood group B glycosyltransferase, which catalyzes the transfer of galactose from UDP‐Gal to Fucα(l‐2)Galβ‐OR to give the blood group B determinant Galα(l‐3)[Fucα(1‐2)]Gal β‐OR (where R is a glycoprotein or glycolipid), has been expressed in Escherichia coli by replacing its membrane‐anchoring domain with an omp A bacterial secretory signal. The active enzyme was purified from the periplasm using UDP‐hexanolamine affinity chromatography and used in the synthesis of preparative amounts of the human blood group B trisaccharide antigen. The substrate specificity and kinetics of the recombinant enzyme were comparable to the enzyme from human sera. Thus we have achieved the construction of a completely synthetic glycosyltransferase gene and its successful expression.</abstract><cop>Oxford, UK</cop><pub>Blackwell Science Ltd</pub><pmid>8529660</pmid><doi>10.1111/j.1432-1033.1995.323_c.x</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | European journal of biochemistry, 1995-11, Vol.234 (1), p.323-328 |
| issn | 0014-2956 1432-1033 |
| language | eng |
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| source | MEDLINE; Alma/SFX Local Collection |
| subjects | ABO Blood-Group System Amino Acid Sequence Base Sequence blood group antigen Carbohydrate Sequence Cloning, Molecular DNA, Recombinant enzymic synthesis Escherichia coli - genetics Galactosyltransferases - genetics Genes, Synthetic glycosyltransferases Humans Kinetics Molecular Sequence Data recombinant Recombinant Proteins - genetics Recombinant Proteins - metabolism synthetic genes Trisaccharides - biosynthesis |
| title | Expression of a Recombinant Human Glycosyltransferase from a Synthetic Gene and its Utilization for Synthesis of the Human Blood Group B Trisaccharide |
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