PROIMI   05436
PLANTA PILOTO DE PROCESOS INDUSTRIALES MICROBIOLOGICOS
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
Characterization of Two Glycosyl Hydrolases from Shewanella sp. G5.
Autor/es:
CRISTÓBAL H. A.; BRECCIA J. D.; ABATE C. M.
Lugar:
Rosario
Reunión:
Congreso; V Congreso Argentino de Microbiología General; 2008
Institución organizadora:
SAMIGE
Resumen:
b-Glucosidases (EC 3.2.1.21) are a heterogeneous group of
enzymes with a broad substrate specificity range over different
aryl- and alkyl-b-D-glucosides. A Shewanella sp. G5 strain was
isolate from the intestinal content of Munida subrrugosa in the
Beagle Channel, Tierra del Fuego (Argentina); this marine
bacteria was able to grow at 4 and 20 ºC using cellobiose as
carbon source. The enzyme was quantified and expressed in UI
(mmol min-1) from assays that determined the temperature
effect at 5, 10, 15, 25, 30, 37 and 45 ºC and pH effect at 3, 4,
5, 6, 7, 8 and 9; using p-Nitrophenyl-b-D-glucopyranoside
(pNPG, Sigma). Zymograms assays were performed using a
fluorescent substrate 4-methylumbelliferyl-b-D-glucopiranoside
(4-MUbG, Sigma). A 100% and 80% of relative activity value
were obtained at 37 ºC, pH 8; and 25 ºC, pH 6, respectively.
Zymograms assays using 4-MUbG showed the presence of two
enzymes, with molecular weight about of 70 and 114 kDa
respectively. These two b-glucosidases were characterized
molecularly in previous studies by the sequencing of two
encoding genes.-Glucosidases (EC 3.2.1.21) are a heterogeneous group of
enzymes with a broad substrate specificity range over different
aryl- and alkyl-b-D-glucosides. A Shewanella sp. G5 strain was
isolate from the intestinal content of Munida subrrugosa in the
Beagle Channel, Tierra del Fuego (Argentina); this marine
bacteria was able to grow at 4 and 20 ºC using cellobiose as
carbon source. The enzyme was quantified and expressed in UI
(mmol min-1) from assays that determined the temperature
effect at 5, 10, 15, 25, 30, 37 and 45 ºC and pH effect at 3, 4,
5, 6, 7, 8 and 9; using p-Nitrophenyl-b-D-glucopyranoside
(pNPG, Sigma). Zymograms assays were performed using a
fluorescent substrate 4-methylumbelliferyl-b-D-glucopiranoside
(4-MUbG, Sigma). A 100% and 80% of relative activity value
were obtained at 37 ºC, pH 8; and 25 ºC, pH 6, respectively.
Zymograms assays using 4-MUbG showed the presence of two
enzymes, with molecular weight about of 70 and 114 kDa
respectively. These two b-glucosidases were characterized
molecularly in previous studies by the sequencing of two
encoding genes.b-D-glucosides. A Shewanella sp. G5 strain was
isolate from the intestinal content of Munida subrrugosa in the
Beagle Channel, Tierra del Fuego (Argentina); this marine
bacteria was able to grow at 4 and 20 ºC using cellobiose as
carbon source. The enzyme was quantified and expressed in UI
(mmol min-1) from assays that determined the temperature
effect at 5, 10, 15, 25, 30, 37 and 45 ºC and pH effect at 3, 4,
5, 6, 7, 8 and 9; using p-Nitrophenyl-b-D-glucopyranoside
(pNPG, Sigma). Zymograms assays were performed using a
fluorescent substrate 4-methylumbelliferyl-b-D-glucopiranoside
(4-MUbG, Sigma). A 100% and 80% of relative activity value
were obtained at 37 ºC, pH 8; and 25 ºC, pH 6, respectively.
Zymograms assays using 4-MUbG showed the presence of two
enzymes, with molecular weight about of 70 and 114 kDa
respectively. These two b-glucosidases were characterized
molecularly in previous studies by the sequencing of two
encoding genes.Munida subrrugosa in the
Beagle Channel, Tierra del Fuego (Argentina); this marine
bacteria was able to grow at 4 and 20 ºC using cellobiose as
carbon source. The enzyme was quantified and expressed in UI
(mmol min-1) from assays that determined the temperature
effect at 5, 10, 15, 25, 30, 37 and 45 ºC and pH effect at 3, 4,
5, 6, 7, 8 and 9; using p-Nitrophenyl-b-D-glucopyranoside
(pNPG, Sigma). Zymograms assays were performed using a
fluorescent substrate 4-methylumbelliferyl-b-D-glucopiranoside
(4-MUbG, Sigma). A 100% and 80% of relative activity value
were obtained at 37 ºC, pH 8; and 25 ºC, pH 6, respectively.
Zymograms assays using 4-MUbG showed the presence of two
enzymes, with molecular weight about of 70 and 114 kDa
respectively. These two b-glucosidases were characterized
molecularly in previous studies by the sequencing of two
encoding genes.mmol min-1) from assays that determined the temperature
effect at 5, 10, 15, 25, 30, 37 and 45 ºC and pH effect at 3, 4,
5, 6, 7, 8 and 9; using p-Nitrophenyl-b-D-glucopyranoside
(pNPG, Sigma). Zymograms assays were performed using a
fluorescent substrate 4-methylumbelliferyl-b-D-glucopiranoside
(4-MUbG, Sigma). A 100% and 80% of relative activity value
were obtained at 37 ºC, pH 8; and 25 ºC, pH 6, respectively.
Zymograms assays using 4-MUbG showed the presence of two
enzymes, with molecular weight about of 70 and 114 kDa
respectively. These two b-glucosidases were characterized
molecularly in previous studies by the sequencing of two
encoding genes.b-D-glucopyranoside
(pNPG, Sigma). Zymograms assays were performed using a
fluorescent substrate 4-methylumbelliferyl-b-D-glucopiranoside
(4-MUbG, Sigma). A 100% and 80% of relative activity value
were obtained at 37 ºC, pH 8; and 25 ºC, pH 6, respectively.
Zymograms assays using 4-MUbG showed the presence of two
enzymes, with molecular weight about of 70 and 114 kDa
respectively. These two b-glucosidases were characterized
molecularly in previous studies by the sequencing of two
encoding genes.b-D-glucopiranoside
(4-MUbG, Sigma). A 100% and 80% of relative activity value
were obtained at 37 ºC, pH 8; and 25 ºC, pH 6, respectively.
Zymograms assays using 4-MUbG showed the presence of two
enzymes, with molecular weight about of 70 and 114 kDa
respectively. These two b-glucosidases were characterized
molecularly in previous studies by the sequencing of two
encoding genes.bG, Sigma). A 100% and 80% of relative activity value
were obtained at 37 ºC, pH 8; and 25 ºC, pH 6, respectively.
Zymograms assays using 4-MUbG showed the presence of two
enzymes, with molecular weight about of 70 and 114 kDa
respectively. These two b-glucosidases were characterized
molecularly in previous studies by the sequencing of two
encoding genes.bG showed the presence of two
enzymes, with molecular weight about of 70 and 114 kDa
respectively. These two b-glucosidases were characterized
molecularly in previous studies by the sequencing of two
encoding genes.b-glucosidases were characterized
molecularly in previous studies by the sequencing of two
encoding genes.