Role of aspergillus niger acra in arsenic resistance and its use as the basis for an arsenic biosensor

Se In Choe, Fabrice N. Gravelat, Qusai Al Abdallah, Mark J. Lee, Bernard F. Gibbs, Donald C. Sheppard

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Arsenic contamination of groundwater sources is a major issue worldwide, since exposure to high levels of arsenic has been linked to a variety of health problems. Effective methods of detection are thus greatly needed as preventive measures. In an effort to develop a fungal biosensor for arsenic, we first identified seven putative arsenic metabolism and transport genes in Aspergillus niger, a widely used industrial organism that is generally regarded as safe (GRAS). Among the genes tested for RNA expression in response to arsenate, acrA, encoding a putative plasma membrane arsenite efflux pump, displayed an over 200-fold increase in gene expression in response to arsenate. We characterized the function of this A. niger protein in arsenic efflux by gene knockout and confirmed that AcrA was located at the cell membrane using an enhanced green fluorescent protein (eGFP) fusion construct. Based on our observations, we developed a putative biosensor strain containing a construct of the native promoter of acrA fused with egfp. We analyzed the fluorescence of this biosensor strain in the presence of arsenic using confocal microscopy and spectrofluorimetry. The biosensor strain reliably detected both arsenite and arsenate in the range of 1.8 to 180 μg/liter, which encompasses the threshold concentrations for drinking water set by the World Health Organization (10 and 50 μg/liter).

Original languageEnglish (US)
Pages (from-to)3855-3863
Number of pages9
JournalApplied and Environmental Microbiology
Volume78
Issue number11
DOIs
StatePublished - Jun 1 2012

Fingerprint

Aspergillus niger
biosensors
Arsenic
Biosensing Techniques
arsenic
arsenates
arsenate
arsenites
arsenite
gene
Cell Membrane
membrane
Gene Knockout Techniques
protein
groundwater contamination
Groundwater
World Health Organization
gene targeting
green fluorescent protein
Confocal Microscopy

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Food Science
  • Applied Microbiology and Biotechnology
  • Ecology

Cite this

Role of aspergillus niger acra in arsenic resistance and its use as the basis for an arsenic biosensor. / Choe, Se In; Gravelat, Fabrice N.; Al Abdallah, Qusai; Lee, Mark J.; Gibbs, Bernard F.; Sheppard, Donald C.

In: Applied and Environmental Microbiology, Vol. 78, No. 11, 01.06.2012, p. 3855-3863.

Research output: Contribution to journalArticle

Choe, Se In ; Gravelat, Fabrice N. ; Al Abdallah, Qusai ; Lee, Mark J. ; Gibbs, Bernard F. ; Sheppard, Donald C. / Role of aspergillus niger acra in arsenic resistance and its use as the basis for an arsenic biosensor. In: Applied and Environmental Microbiology. 2012 ; Vol. 78, No. 11. pp. 3855-3863.
@article{525322945aff45f4a9e307b753ba38a6,
title = "Role of aspergillus niger acra in arsenic resistance and its use as the basis for an arsenic biosensor",
abstract = "Arsenic contamination of groundwater sources is a major issue worldwide, since exposure to high levels of arsenic has been linked to a variety of health problems. Effective methods of detection are thus greatly needed as preventive measures. In an effort to develop a fungal biosensor for arsenic, we first identified seven putative arsenic metabolism and transport genes in Aspergillus niger, a widely used industrial organism that is generally regarded as safe (GRAS). Among the genes tested for RNA expression in response to arsenate, acrA, encoding a putative plasma membrane arsenite efflux pump, displayed an over 200-fold increase in gene expression in response to arsenate. We characterized the function of this A. niger protein in arsenic efflux by gene knockout and confirmed that AcrA was located at the cell membrane using an enhanced green fluorescent protein (eGFP) fusion construct. Based on our observations, we developed a putative biosensor strain containing a construct of the native promoter of acrA fused with egfp. We analyzed the fluorescence of this biosensor strain in the presence of arsenic using confocal microscopy and spectrofluorimetry. The biosensor strain reliably detected both arsenite and arsenate in the range of 1.8 to 180 μg/liter, which encompasses the threshold concentrations for drinking water set by the World Health Organization (10 and 50 μg/liter).",
author = "Choe, {Se In} and Gravelat, {Fabrice N.} and {Al Abdallah}, Qusai and Lee, {Mark J.} and Gibbs, {Bernard F.} and Sheppard, {Donald C.}",
year = "2012",
month = "6",
day = "1",
doi = "10.1128/AEM.07771-11",
language = "English (US)",
volume = "78",
pages = "3855--3863",
journal = "Applied and Environmental Microbiology",
issn = "0099-2240",
publisher = "American Society for Microbiology",
number = "11",

}

TY - JOUR

T1 - Role of aspergillus niger acra in arsenic resistance and its use as the basis for an arsenic biosensor

AU - Choe, Se In

AU - Gravelat, Fabrice N.

AU - Al Abdallah, Qusai

AU - Lee, Mark J.

AU - Gibbs, Bernard F.

AU - Sheppard, Donald C.

PY - 2012/6/1

Y1 - 2012/6/1

N2 - Arsenic contamination of groundwater sources is a major issue worldwide, since exposure to high levels of arsenic has been linked to a variety of health problems. Effective methods of detection are thus greatly needed as preventive measures. In an effort to develop a fungal biosensor for arsenic, we first identified seven putative arsenic metabolism and transport genes in Aspergillus niger, a widely used industrial organism that is generally regarded as safe (GRAS). Among the genes tested for RNA expression in response to arsenate, acrA, encoding a putative plasma membrane arsenite efflux pump, displayed an over 200-fold increase in gene expression in response to arsenate. We characterized the function of this A. niger protein in arsenic efflux by gene knockout and confirmed that AcrA was located at the cell membrane using an enhanced green fluorescent protein (eGFP) fusion construct. Based on our observations, we developed a putative biosensor strain containing a construct of the native promoter of acrA fused with egfp. We analyzed the fluorescence of this biosensor strain in the presence of arsenic using confocal microscopy and spectrofluorimetry. The biosensor strain reliably detected both arsenite and arsenate in the range of 1.8 to 180 μg/liter, which encompasses the threshold concentrations for drinking water set by the World Health Organization (10 and 50 μg/liter).

AB - Arsenic contamination of groundwater sources is a major issue worldwide, since exposure to high levels of arsenic has been linked to a variety of health problems. Effective methods of detection are thus greatly needed as preventive measures. In an effort to develop a fungal biosensor for arsenic, we first identified seven putative arsenic metabolism and transport genes in Aspergillus niger, a widely used industrial organism that is generally regarded as safe (GRAS). Among the genes tested for RNA expression in response to arsenate, acrA, encoding a putative plasma membrane arsenite efflux pump, displayed an over 200-fold increase in gene expression in response to arsenate. We characterized the function of this A. niger protein in arsenic efflux by gene knockout and confirmed that AcrA was located at the cell membrane using an enhanced green fluorescent protein (eGFP) fusion construct. Based on our observations, we developed a putative biosensor strain containing a construct of the native promoter of acrA fused with egfp. We analyzed the fluorescence of this biosensor strain in the presence of arsenic using confocal microscopy and spectrofluorimetry. The biosensor strain reliably detected both arsenite and arsenate in the range of 1.8 to 180 μg/liter, which encompasses the threshold concentrations for drinking water set by the World Health Organization (10 and 50 μg/liter).

UR - http://www.scopus.com/inward/record.url?scp=84864099499&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84864099499&partnerID=8YFLogxK

U2 - 10.1128/AEM.07771-11

DO - 10.1128/AEM.07771-11

M3 - Article

VL - 78

SP - 3855

EP - 3863

JO - Applied and Environmental Microbiology

JF - Applied and Environmental Microbiology

SN - 0099-2240

IS - 11

ER -