Characterization of Two Glycosyl Hydrolases from Shewanella sp. G5.

CRISTÓBAL H. A.; BRECCIA J. D.; ABATE C. M.

Rosario

Congreso; V Congreso Argentino de Microbiología General; 2008

SAMIGE

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.