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Expression and applications of HriCFP in E. coli: a novel biosensing fluorescent protein

Expression and applications of HriCFP in E. coli: a novel biosensing fluorescent protein
Expression and applications of HriCFP in E. coli: a novel biosensing fluorescent protein
Metalloids and heavy metal contamination in the environment has become a global problem. Therefore, there is a dire need to develop effective and inexpensive approaches that can facilitate efficient monitoring of the hazardous level of these environmental pollutants. Microbial cell-based and fluorescent protein-based biosensors offer relatively convenient and inexpensive tools for the analysis of environmental pollutants as opposed to traditional instrumental approaches. Small size fluorescent proteins can withstand exposure to denaturants, high temperature and a wide pH range. These characteristics, along with their potential of sensing different toxic analytes, makes them a suitable candidate for developing on-site detection biosensors. The current study exploits the biosensing potential of a novel fluorescent protein called HriCFP. HriCFP was expressed in the prokaryotic system (gram-negative E. coli), which showed stable and discreet expression in bacterial cells. Whole-cell biosensors (WCB) were developed by immobilization of HriCFP expressing non-pathogenic E. coli via nitrocellulose membrane, low melting agarose and sodium silicate gel. These immobilized biosensors were tested for their sensitivity of detection for environmental pollutants, i.e., heavy metals (Cu(II), Hg(II), As(III)). These WCBs exhibited profound fluorescent quenching when exposed to a range of heavy metals. These biosensors remained active for 12 days at 4 °C, demonstrating their potential for long-term stability and storage. This study implies that HriCFP may have a significant advantage over other larger and multimeric proteins as it has a minimal impact on host strain metabolism and hence, increasing its sustainability for a longer period.
1948-5948
Mehreen, Hira
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Saeed, Salma
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Gerlevik, Umut
2f10731e-aca9-473f-a474-a37b74855c2f
Tariq, Aamira
4f936c30-424c-4cf6-8f95-cb7e81866140
Sezerman, Ugur
caf306fc-5ec1-4432-8bc8-2ed45799a566
Noreen, Zobia
60abc4bc-f93a-42f7-9703-e75e90b9e953
Zhang, Xunli
d7cf1181-3276-4da1-9150-e212b333abb1
Hassan, Sammer-Ul
8a5ae3f1-3451-4093-879e-85f40953da8b
Bokhari, habib
1c3a1e18-4570-4a10-9feb-cea4218a275f
Mehreen, Hira
f835a3ac-15e5-4347-b8bb-d4826d2c463b
Saeed, Salma
5ccd0720-fdd2-4176-890e-b400aa5811c8
Gerlevik, Umut
2f10731e-aca9-473f-a474-a37b74855c2f
Tariq, Aamira
4f936c30-424c-4cf6-8f95-cb7e81866140
Sezerman, Ugur
caf306fc-5ec1-4432-8bc8-2ed45799a566
Noreen, Zobia
60abc4bc-f93a-42f7-9703-e75e90b9e953
Zhang, Xunli
d7cf1181-3276-4da1-9150-e212b333abb1
Hassan, Sammer-Ul
8a5ae3f1-3451-4093-879e-85f40953da8b
Bokhari, habib
1c3a1e18-4570-4a10-9feb-cea4218a275f

Mehreen, Hira, Saeed, Salma, Gerlevik, Umut, Tariq, Aamira, Sezerman, Ugur, Noreen, Zobia, Zhang, Xunli, Hassan, Sammer-Ul and Bokhari, habib (2020) Expression and applications of HriCFP in E. coli: a novel biosensing fluorescent protein. Journal of Microbial and Biochemical Technology, [434]. (In Press)

Record type: Article

Abstract

Metalloids and heavy metal contamination in the environment has become a global problem. Therefore, there is a dire need to develop effective and inexpensive approaches that can facilitate efficient monitoring of the hazardous level of these environmental pollutants. Microbial cell-based and fluorescent protein-based biosensors offer relatively convenient and inexpensive tools for the analysis of environmental pollutants as opposed to traditional instrumental approaches. Small size fluorescent proteins can withstand exposure to denaturants, high temperature and a wide pH range. These characteristics, along with their potential of sensing different toxic analytes, makes them a suitable candidate for developing on-site detection biosensors. The current study exploits the biosensing potential of a novel fluorescent protein called HriCFP. HriCFP was expressed in the prokaryotic system (gram-negative E. coli), which showed stable and discreet expression in bacterial cells. Whole-cell biosensors (WCB) were developed by immobilization of HriCFP expressing non-pathogenic E. coli via nitrocellulose membrane, low melting agarose and sodium silicate gel. These immobilized biosensors were tested for their sensitivity of detection for environmental pollutants, i.e., heavy metals (Cu(II), Hg(II), As(III)). These WCBs exhibited profound fluorescent quenching when exposed to a range of heavy metals. These biosensors remained active for 12 days at 4 °C, demonstrating their potential for long-term stability and storage. This study implies that HriCFP may have a significant advantage over other larger and multimeric proteins as it has a minimal impact on host strain metabolism and hence, increasing its sustainability for a longer period.

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1948-5948-12-434[5211] - Accepted Manuscript
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Accepted/In Press date: 15 July 2020

Identifiers

Local EPrints ID: 442827
URI: http://eprints.soton.ac.uk/id/eprint/442827
ISSN: 1948-5948
PURE UUID: 02910c0b-76bb-4f64-ab58-86d6f6ae29fd
ORCID for Xunli Zhang: ORCID iD orcid.org/0000-0002-4375-1571
ORCID for Sammer-Ul Hassan: ORCID iD orcid.org/0000-0002-0319-5814

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Date deposited: 28 Jul 2020 16:31
Last modified: 07 Oct 2020 02:02

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