Artículo

Geremia, R.A.; Roux, M.; Ferreiro, D.U.; Dauphin-Dubois, R.; Lellouch, A.C.; Ielpi, L. "Expression and biochemical characterisation of recombinant AceA, a bacterial α-mannosyltransferase" (1999) Molecular and General Genetics. 261(6):933-940
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Abstract:

Biosynthesis of repeat-unit polysaccharides and N-linked glycans proceeds by sequential transfer of sugars from the appropriate sugar donor to an activated lipid carrier. The transfer of each sugar is catalysed by a specific glycosyltransferase. The molecular basis of the specificity of sugar addition is not yet well understood, mainly because of the difficulty of isolating these proteins. In this study, the aceA gene product expressed by Acetobacter xylinum, which is involved in the biosynthesis of the exopolysaccharide acetan, was overproduced in Escherichia coli and its function was characterised. The aceA ORF was subcloned into the expression vector pET29 in frame with the S·tag epitope. The recombinant protein was identified, and culture conditions were optimised for production of the soluble protein. The results of test reactions showed that AceA is able to transfer one α-mannose residue from GDP-mannose to cellobiose-P-P-lipid to produce α-mannose-cellobiose-P-P-lipid. AceA was not able to use free cellobiose as a substrate, indicating that the pyrophosphate-lipid moiety is needed for enzymatic activity.

Registro:

Documento: Artículo
Título:Expression and biochemical characterisation of recombinant AceA, a bacterial α-mannosyltransferase
Autor:Geremia, R.A.; Roux, M.; Ferreiro, D.U.; Dauphin-Dubois, R.; Lellouch, A.C.; Ielpi, L.
Filiación:Ctr. Rech. sur les Macromolec. Veg., CNRS, Université Joseph Fourier, BP 53X, F-38041 Grenoble Cedex 9, France
Inst. Invest. Bioquim. Fund. C., Fac. de Ciencias Exactas y Naturales, Consejo Nac. de Invest. Cie./Tec., Avda. Patricias Argentinas 451, 1045 Buenos Aires, Argentina
Palabras clave:Acetan; Acetobacter xylinum; Bacterial polysaccharide; Biosynthesis; Glycosyltransferase; bacterial enzyme; cellobiose; guanosine diphosphate mannose; mannose; mannosyltransferase; recombinant enzyme; acetobacter; article; controlled study; enzyme activity; escherichia coli; expression vector; gene overexpression; molecular cloning; nonhuman; open reading frame; priority journal; structure activity relation; Amino Acid Sequence; Base Sequence; Carbohydrate Sequence; Cloning, Molecular; DNA, Recombinant; Mannosyltransferases; Mutagenesis, Site-Directed; Open Reading Frames; Substrate Specificity; Acetobacter; Bacteria (microorganisms); Escherichia coli; Gluconacetobacter xylinus
Año:1999
Volumen:261
Número:6
Página de inicio:933
Página de fin:940
DOI: http://dx.doi.org/10.1007/s004380051040
Título revista:Molecular and General Genetics
Título revista abreviado:Mol. Gen. Genet.
ISSN:00268925
CODEN:MGGEA
CAS:DNA, Recombinant; Mannosyltransferases, EC 2.4.1.-
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00268925_v261_n6_p933_Geremia

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Citas:

---------- APA ----------
Geremia, R.A., Roux, M., Ferreiro, D.U., Dauphin-Dubois, R., Lellouch, A.C. & Ielpi, L. (1999) . Expression and biochemical characterisation of recombinant AceA, a bacterial α-mannosyltransferase. Molecular and General Genetics, 261(6), 933-940.
http://dx.doi.org/10.1007/s004380051040
---------- CHICAGO ----------
Geremia, R.A., Roux, M., Ferreiro, D.U., Dauphin-Dubois, R., Lellouch, A.C., Ielpi, L. "Expression and biochemical characterisation of recombinant AceA, a bacterial α-mannosyltransferase" . Molecular and General Genetics 261, no. 6 (1999) : 933-940.
http://dx.doi.org/10.1007/s004380051040
---------- MLA ----------
Geremia, R.A., Roux, M., Ferreiro, D.U., Dauphin-Dubois, R., Lellouch, A.C., Ielpi, L. "Expression and biochemical characterisation of recombinant AceA, a bacterial α-mannosyltransferase" . Molecular and General Genetics, vol. 261, no. 6, 1999, pp. 933-940.
http://dx.doi.org/10.1007/s004380051040
---------- VANCOUVER ----------
Geremia, R.A., Roux, M., Ferreiro, D.U., Dauphin-Dubois, R., Lellouch, A.C., Ielpi, L. Expression and biochemical characterisation of recombinant AceA, a bacterial α-mannosyltransferase. Mol. Gen. Genet. 1999;261(6):933-940.
http://dx.doi.org/10.1007/s004380051040