Cargando…
A Biosensor Platform for Metal Detection Based on Enhanced Green Fluorescent Protein
Microbial cell-based biosensors, which mostly rely on stress-responsive operons, have been widely developed to monitor environmental pollutants. Biosensors are usually more convenient and inexpensive than traditional instrumental analyses of environmental pollutants. However, the targets of biosenso...
Autores principales: | , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2019
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6514868/ https://www.ncbi.nlm.nih.gov/pubmed/31003435 http://dx.doi.org/10.3390/s19081846 |
_version_ | 1783417960089518080 |
---|---|
author | Lee, Woonwoo Kim, Hyojin Kang, Yerin Lee, Youngshim Yoon, Youngdae |
author_facet | Lee, Woonwoo Kim, Hyojin Kang, Yerin Lee, Youngshim Yoon, Youngdae |
author_sort | Lee, Woonwoo |
collection | PubMed |
description | Microbial cell-based biosensors, which mostly rely on stress-responsive operons, have been widely developed to monitor environmental pollutants. Biosensors are usually more convenient and inexpensive than traditional instrumental analyses of environmental pollutants. However, the targets of biosensors are restricted by the limited number of genetic operon systems available. In this study, we demonstrated a novel strategy to overcome this limitation by engineering an enhanced green fluorescent protein (eGFP). It has been reported that combining two fragments of split-eGFP can form a native structure. Thus, we engineered new biosensors by inserting metal-binding loops (MBLs) between β-strands 9 and 10 of the eGFP, which then undergoes conformational changes upon interaction between the MBLs and targets, thereby emitting fluorescence. The two designed MLBs based on our previous study were employed as linkers between two fragments of eGFP. As a result, an Escherichia coli biosensor exhibited a fluorescent signal only when interacting with cadmium ions, revealing the prospect of a new biosensor for cadmium detection. Although this study is a starting stage for further developing biosensors, we believe that the proposed strategy can serve as basis to develop new biosensors to target various environmental pollutants. |
format | Online Article Text |
id | pubmed-6514868 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-65148682019-05-30 A Biosensor Platform for Metal Detection Based on Enhanced Green Fluorescent Protein Lee, Woonwoo Kim, Hyojin Kang, Yerin Lee, Youngshim Yoon, Youngdae Sensors (Basel) Article Microbial cell-based biosensors, which mostly rely on stress-responsive operons, have been widely developed to monitor environmental pollutants. Biosensors are usually more convenient and inexpensive than traditional instrumental analyses of environmental pollutants. However, the targets of biosensors are restricted by the limited number of genetic operon systems available. In this study, we demonstrated a novel strategy to overcome this limitation by engineering an enhanced green fluorescent protein (eGFP). It has been reported that combining two fragments of split-eGFP can form a native structure. Thus, we engineered new biosensors by inserting metal-binding loops (MBLs) between β-strands 9 and 10 of the eGFP, which then undergoes conformational changes upon interaction between the MBLs and targets, thereby emitting fluorescence. The two designed MLBs based on our previous study were employed as linkers between two fragments of eGFP. As a result, an Escherichia coli biosensor exhibited a fluorescent signal only when interacting with cadmium ions, revealing the prospect of a new biosensor for cadmium detection. Although this study is a starting stage for further developing biosensors, we believe that the proposed strategy can serve as basis to develop new biosensors to target various environmental pollutants. MDPI 2019-04-18 /pmc/articles/PMC6514868/ /pubmed/31003435 http://dx.doi.org/10.3390/s19081846 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Lee, Woonwoo Kim, Hyojin Kang, Yerin Lee, Youngshim Yoon, Youngdae A Biosensor Platform for Metal Detection Based on Enhanced Green Fluorescent Protein |
title | A Biosensor Platform for Metal Detection Based on Enhanced Green Fluorescent Protein |
title_full | A Biosensor Platform for Metal Detection Based on Enhanced Green Fluorescent Protein |
title_fullStr | A Biosensor Platform for Metal Detection Based on Enhanced Green Fluorescent Protein |
title_full_unstemmed | A Biosensor Platform for Metal Detection Based on Enhanced Green Fluorescent Protein |
title_short | A Biosensor Platform for Metal Detection Based on Enhanced Green Fluorescent Protein |
title_sort | biosensor platform for metal detection based on enhanced green fluorescent protein |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6514868/ https://www.ncbi.nlm.nih.gov/pubmed/31003435 http://dx.doi.org/10.3390/s19081846 |
work_keys_str_mv | AT leewoonwoo abiosensorplatformformetaldetectionbasedonenhancedgreenfluorescentprotein AT kimhyojin abiosensorplatformformetaldetectionbasedonenhancedgreenfluorescentprotein AT kangyerin abiosensorplatformformetaldetectionbasedonenhancedgreenfluorescentprotein AT leeyoungshim abiosensorplatformformetaldetectionbasedonenhancedgreenfluorescentprotein AT yoonyoungdae abiosensorplatformformetaldetectionbasedonenhancedgreenfluorescentprotein AT leewoonwoo biosensorplatformformetaldetectionbasedonenhancedgreenfluorescentprotein AT kimhyojin biosensorplatformformetaldetectionbasedonenhancedgreenfluorescentprotein AT kangyerin biosensorplatformformetaldetectionbasedonenhancedgreenfluorescentprotein AT leeyoungshim biosensorplatformformetaldetectionbasedonenhancedgreenfluorescentprotein AT yoonyoungdae biosensorplatformformetaldetectionbasedonenhancedgreenfluorescentprotein |