INSIBIO   05451
INSTITUTO SUPERIOR DE INVESTIGACIONES BIOLOGICAS
Unidad Ejecutora - UE
artículos
Título:
Copper bioaccumulation by the actinobacterium Amycolatopsis sp
Autor/es:
ALBARRACÍN VH; WINIK BC; KOTHE E; AMOROSO MJ; ABATE CM
Revista:
JOURNAL OF BASIC MICROBIOLOGY
Editorial:
Wiley-VCH Verlag
Referencias:
Lugar: Weinheim - Alemania; Año: 2008 vol. 48 p. 1 - 8
ISSN:
0233-111X
Resumen:
Amycolatopsis sp. AB0, a copper resistant actinobacterium isolated from polluted sediments, has
shown high copper specific biopsortion ability (25 mg g1). Two approaches were used to
confirm metal accumulation in growing cells of Amycolatopsis sp. AB0; we performed
subcellular fractioning assays which showed that the retained copper was associated with the
extra-cellular fraction (exopolymer, 40%), but mainly within the cells. Intracellular
distribution of copper was: 86% in the cytosolic fraction, 11% at the cell wall and 3%
associated with the ribosome/membrane fraction. Its copper bioaccumulation ability was
corroborated by using silver enhanced staining of copper with the Timms reagent technique,
which has not been used to detect metal deposits in bacteria before. In addition, we
constructed specific oligonucleotides for targeting genes coding for copper P-Type ATPases that
could be involved in the copper uptake ability of this strain. A 607 bp DNA fragment was
amplified and sequenced from Amycolatopsis sp AB0. BLAST search analysis showed 71% protein
homology of the deduced sequence with a putative cation-transporting ATPase of Nocardia
farcinica and 65% with a copper translocating ATPase of Mycobacterium flavescens. To our
knowledge this is the first report of the presence of copper P-type ATPase genes in thesp. AB0, a copper resistant actinobacterium isolated from polluted sediments, has
shown high copper specific biopsortion ability (25 mg g1). Two approaches were used to
confirm metal accumulation in growing cells of Amycolatopsis sp. AB0; we performed
subcellular fractioning assays which showed that the retained copper was associated with the
extra-cellular fraction (exopolymer, 40%), but mainly within the cells. Intracellular
distribution of copper was: 86% in the cytosolic fraction, 11% at the cell wall and 3%
associated with the ribosome/membrane fraction. Its copper bioaccumulation ability was
corroborated by using silver enhanced staining of copper with the Timms reagent technique,
which has not been used to detect metal deposits in bacteria before. In addition, we
constructed specific oligonucleotides for targeting genes coding for copper P-Type ATPases that
could be involved in the copper uptake ability of this strain. A 607 bp DNA fragment was
amplified and sequenced from Amycolatopsis sp AB0. BLAST search analysis showed 71% protein
homology of the deduced sequence with a putative cation-transporting ATPase of Nocardia
farcinica and 65% with a copper translocating ATPase of Mycobacterium flavescens. To our
knowledge this is the first report of the presence of copper P-type ATPase genes in the1). Two approaches were used to
confirm metal accumulation in growing cells of Amycolatopsis sp. AB0; we performed
subcellular fractioning assays which showed that the retained copper was associated with the
extra-cellular fraction (exopolymer, 40%), but mainly within the cells. Intracellular
distribution of copper was: 86% in the cytosolic fraction, 11% at the cell wall and 3%
associated with the ribosome/membrane fraction. Its copper bioaccumulation ability was
corroborated by using silver enhanced staining of copper with the Timms reagent technique,
which has not been used to detect metal deposits in bacteria before. In addition, we
constructed specific oligonucleotides for targeting genes coding for copper P-Type ATPases that
could be involved in the copper uptake ability of this strain. A 607 bp DNA fragment was
amplified and sequenced from Amycolatopsis sp AB0. BLAST search analysis showed 71% protein
homology of the deduced sequence with a putative cation-transporting ATPase of Nocardia
farcinica and 65% with a copper translocating ATPase of Mycobacterium flavescens. To our
knowledge this is the first report of the presence of copper P-type ATPase genes in theAmycolatopsis sp. AB0; we performed
subcellular fractioning assays which showed that the retained copper was associated with the
extra-cellular fraction (exopolymer, 40%), but mainly within the cells. Intracellular
distribution of copper was: 86% in the cytosolic fraction, 11% at the cell wall and 3%
associated with the ribosome/membrane fraction. Its copper bioaccumulation ability was
corroborated by using silver enhanced staining of copper with the Timms reagent technique,
which has not been used to detect metal deposits in bacteria before. In addition, we
constructed specific oligonucleotides for targeting genes coding for copper P-Type ATPases that
could be involved in the copper uptake ability of this strain. A 607 bp DNA fragment was
amplified and sequenced from Amycolatopsis sp AB0. BLAST search analysis showed 71% protein
homology of the deduced sequence with a putative cation-transporting ATPase of Nocardia
farcinica and 65% with a copper translocating ATPase of Mycobacterium flavescens. To our
knowledge this is the first report of the presence of copper P-type ATPase genes in theAmycolatopsis sp AB0. BLAST search analysis showed 71% protein
homology of the deduced sequence with a putative cation-transporting ATPase of Nocardia
farcinica and 65% with a copper translocating ATPase of Mycobacterium flavescens. To our
knowledge this is the first report of the presence of copper P-type ATPase genes in theNocardia
farcinica and 65% with a copper translocating ATPase of Mycobacterium flavescens. To our
knowledge this is the first report of the presence of copper P-type ATPase genes in theand 65% with a copper translocating ATPase of Mycobacterium flavescens. To our
knowledge this is the first report of the presence of copper P-type ATPase genes in the
Amycolotopsis genus.genus.