Microbial biosensors

[ Detection and treatment of soil and water pollutants and organic

explosives]
Contents
1.
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 201
2. Advantages of using microorganisms as biosensing elements . . . . . . . . . . .
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 201
3. Immobilization of
microorganisms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
. . . . . . . . . . . . . . . . . . . . . . .201
3.1. Chemical methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 201
3.2. Physical methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 201
4. Electrochemical microbial
biosensor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 202
4.1. Amperometric microbial
biosensor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
. . . . . . . . . . . . . . . . . . . . . . . . . . . . 202
4.2. Potentiometric microbial biosensor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 204
4.3. Conductimetric
biosensor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 205
4.4. Microbial fuel cell type
biosensor. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 205
5. Optical microbial
biosensor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 205
5.1. Bioluminescence biosensor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 205
5.2. Fluorescence biosensor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 207
5.2.1. Green fluorescence protein-based biosensor. . . . . . . . . . . . . . . . . . . . . .
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 207
5.2.2. O2-sensitive fluorescent material-based
biosensor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
. . . . . . . 207
5.3. Colorimetric biosensor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 207
6. Other types of microbial biosensors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 208
6.1. Sensors based on baroxymeter for the detection of pressure change . . . .
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 208
6.2. Sensors based on infrared analyzer for the detection of the microbial
respiration product CO2 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 208
7. Future
trends . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 208
Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 208
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 208

1-Introduction
A biosensor is an analytical device that combines a biological sensing
element with a transducer to produce a signal proportional to the analyte
concentration . This signal can result from a change in protons concentration,
release or uptake of gases, light emission, absorption and so forth, brought
about by the metabolism of the target compound by the biological
recognition element. The transducer converts this biological signal into a
measurable response such as current, potential or absorption of light through
electrochemical or optical means, which can be further amplified, processed
and stored for later analysis.
A microbial biosensor is an analytical device which integrates
microorganism(s) with a physical transducer to generate a measurable signal
proportional to the concentration of analytes. In recent years, a large number
of microbial biosensors have been developed for environmental, food, and
biomedical applications. Microorganisms have been integrated with a variety
of transducers such as amperometric, potentiometric, calorimetric,
conductimetric, colorimetric, luminescence and fluorescence to construct
biosensor devices. The first reported biosensors based on microorganisms
related the respiration rate of the immobilized microbes to the concentration
of the substrate to be detected. Such biosensors were used for BOD
monitoring. BOD sensors offer a great advantage over the five day BOD
laboratory test. Such biosensors have been also reported for toxicity
monitoring. Canada is a world leader in this highly commercializable
technology. However, the most recent focus on microbial biosensors is based
on the use of recombinant microorganisms that recognize and report the
presence of specific environmental pollutants.

2-Significance of microbial sensors

Environmental issues are most concerned now days. Concern over the
pollution risk to drinking water from industry and agriculture is growing, and
the need for continuous on-line monitoring is there. There is increasing use
of living organisms as the sensitive agent to detect the presence of
pollutants, and whole-cell biosensors are seen to have particular advantages
in such environmental monitoring. The development of a mediated
amperometric biosensor, incorporating the Cyanobacterium synechococcus
as the biocatalyst, for on-line herbicide monitoring is developed. The
biosensor is able to detect a wide range of herbicides with sites of action on
the photosynthetic electron transport chain. Thus microbial biosensors can
help in mitigating problems related to our basic needs i.e. food and water,
hence overcoming many health and environment problems. Microbial
sensors have the advantages of tolerance to measuring conditions, a long
lifetime, and cost performance.
3-Biosensensing

4-Generalized scheme of biosensors

5-History of biosensors:
The first and the most widely used commercial biosensor: the
blood glucose biosensor -developed by Leland C. Clark in
1962.

The first microbial biosensor was described in 1975 by Divies and was
based on the use of Acetobacter xylinum and oxygen electrode. The work
became the foundation of the investigations devoted to the development of
microbial biosensors for application in biotechnology and environmental
monitoring.

6-Advantage of using microbial cells

The use of microbial cells in place of isolated enzymes offers several advantages over enzyme
electrodes, such as,
Elimination of the tedious enzyme extraction and purification steps,
Avoidance of the need for a cofactor, and increased stability. The microbial
sensors show an increased stability because of the enzyme environment is
optimized by evolution and well suited for recovery. These sensors are
essentially living and may be fed and kept alive for a long period..
Furthermore, the whole cell may perform multistep transformations that
could be difficult, if not impossible, to achieve with single enzymes. However,
microbial sensors suffer from the multireceptor behavior of intact cells,
resulting in a rather poor selectivity.

7-Working principle of biosensors

8-Immobilization of microorganisms
The basis of a microbial biosensor is the close contact between
microorganisms and the transducer. Thus, fabrication of a microbial
biosensor requires immobilization on transducers. Microorganisms can be
immobilized on transducer or support matrices by chemical or physical
methods.
8.1-Chemical methods
Chemical methods of microbe immobilization include covalent binding and
cross-linking. Covalent binding methods rely on the formation of a stable
covalent bond between functional groups of the microorganisms cell wall
components such as amine, carboxylic or sulphydryl and the transducer such
as amine, carboxylic, epoxy or tosyl.

8.2-Physical methods
Adsorption and entrapment are the two widely used physical methods for
microbial immobilization. Physical adsorption is the simplest method for
microbe immobilization. Typically, a microbial suspension is incubated with
the electrode or an immobilization matrix, such as alumina and glass bead,
followed by rinsing with buffer to remove unadsorbed cells. The microbes are
immobilized due to adsorptive interactions such as ionic, polar or hydrogen
bonding and hydrophobic interaction.
The immobilization of microorganisms by entrapment can be
achieved by the either retention of the cells in close proximity of
the transducer surface using dialysis or filter membrane or in
chemical/biological polymers/gels such as
(Alginate, carrageenan, agarose, chitosan, collagen,
polyacrylamide, polyvinylachohol, poly(ethylene glycol),
polyurethane,etc.

9- Types of Microbial Biosensors (Transducers)

Microbial biosensor can be classified based on the transducers into
electrochemical, optical and others.

Figure: Design of Microbial sensor 1, isolator; 2,

electrolyte; 3, anode; 4, cathode; 5, Teflon membrane; 6, immobilized microorganism, and
7, dialysis membrane.

9.1-Electrochemical transducers
 The biochemical signals can be used to generate a current/charge or may
change conductivity between two electrodes and therefore the
corresponding transduction device has been described as amperometric,
potentiometric and conductimetric. Each will be briefly described.

9.1.1- Amperometric microbial biosensor

Amperometric microbial biosensor operates at fixed potential with respect to
a reference electrode and involves the detection of the current generated by
the oxidation or reduction of species at the surface of the electrode.
Amperometric microbial biosensors have been widely developed for the
determination of biochemical oxygen demand (BOD) for the
measurement of biodegradable organic pollutants in aqueous samples.
The microbial strains used as biological sensing element include Torulopsis
candida], Trichosporon cutaneum, Pseudomonas putida, Bacillus subtilis ,
Arxula adeninivorans LS3, Serratia marcescens LSY4, Pseudomonas
sp.,Thermophilic bacteria, Hansennula anomal and yeast.
As the most extensively investigated microbial biosensor, the first
commercial BOD biosensor was produced by Nisshin Denki (Electric) in
1983.

Figure 2: Schematic diagram of a simple amperometric biosensor. A potential is applied

between the central platinum cathode and the annular silver anode. This generates a
current (I) which is carried between the electrodes by means of a saturated solution of KCl.
This electrode compartment is separated from the biocatalyst (here shown glucose oxidase)
by a thin plastic membrane, permeable only to oxygen. The analyte solution is separated
from the biocatalyst by another membrane, permeable to the substrate(s) and product(s).
This biosensor is normally about 1 cm in diameter but has been scaled down to 0.25 mm
diameter using a Pt wire cathode within a silver plated steel needle anode and utilising dip-
coated membranes.

9.1.2-Potentiometric microbial biosensor

Conventional potentiometric microbial biosensors consist of an ion-selective electrode (pH,
ammonium, chloride and so on) or a gas-sensing electrode (pCO2 and pNH3 ) coated with an
immobilized microbe layer. Microbe consuming analyte generates a change in potential
resulting from ion accumulation or depletion. Potentiometric transducers measure the
difference between a working electrode and a reference electrode, and the signal is
correlated to the concentration of analyte. Due to a logarithmic relationship between the
potential generated
and analyte concentration, a wide detection range is possible. There are three types of
potentiometric devices: (i) ion-selective electrodes (ISEs), are usually pH meter electrodes, (ii) gas-sensing
electrodes and (iii) field effect transistors (FETs) which are usually solid state electrodes.

Figure 3: A simple potentiometric biosensor. A semi-permeable membrane (a) surrounds the

biocatalyst (b) entrapped next to the active glass membrane (c) of a pH probe (d). The electrical potential (e) is generated
between the internal Ag/AgCl electrode (f) bathed in dilute HCl (g) and an external reference electrode (h).
9.1.3-Conductimetric biosensor
Many microbe-catalyzed reactions involve a change in ionic species. Associated with this
change is a net change in the conductivity of the reaction solution. Even though the
detection of solution conductance is non-specific, conductance measurements are extremely
sensitive.Recently, single-use conductivity and microbial sensor were developed to
investigate the effect of both species and concentration/ osmolarity of anions on the
metabolic activity of E. coli. This hybrid sensing system combines physico-chemical and
biological sensing and greatly increases the ease with which comparative data could be
assimilated.
9.1.4-Microbial fuel cell type biosensor
Microbial fuel cells (MFCs) have been studied as a BOD sensor for a long time. Since Karube
et al. reported a BOD
sensor based on MFC using the hydrogen produced by Clostridium butyricum immobilized on
the electrode in 1977, a variety of MFC BOD sensors with use of electron-mediator have
been developed. Even though the addition of mediators in these biosensors can enhance the
electron transfer, these biosensors have poor stability because of the toxicity of mediators.
9.2-Optical microbial biosensor
The modulation in optical properties such as UVvis absorption, bio- and chemi-
luminescence, reflectance and fluorescence brought by the interaction of the biocatalyst
with the target analyte is the basis for optical microbial biosensors. Opticalbased biosensors
offer advantages of compactness, flexibility, resistance to electrical noise, and a small probe
size.
9.2.1-Bioluminescence biosensor
Bioluminescence is associated with the emission of light by living microorganisms and it
plays a very important role in realtime process monitoring. The bacterial luminescence lux
gene has been widely applied as a reporter either in an inducible or constitutive manner. In
the inducible manner, the reporter lux gene is fused to a promoter regulated by the
concentration of a compound of interest. As a result, the concentration of the compound can
be quantitatively analyzed by detecting the bioluminescence intensity. In the constitutive
manner, the reporter gene is fused to promoters that are continuously expressed as long as
the organism is alive and metabolically active. This kind of reporter is good for evaluating
the total toxicity of contaminant. Both types of reporters have been shown to be useful for
biosensor development. Bioluminescent microbial biosensors have been extensively
investigated to monitor bioavailable metal.
9.2.2-Fluorescence biosensor
Fluorescence spectroscopy has been widely applied in analytical chemistry. It is a sensitive
technique that can detect very low concentrations of analyte because of the instrumental
principles involved. At low analyte concentrations, fluorescence emission intensity is directly
proportional to the concentration. Fluorescent materials and green fluorescent protein have
been extensively used in the construction of fluorescent biosensor.
9.2.2.1-Green fluorescence protein-based biosensor
The GFP-based microbial biosensor has been shown to be useful in assessing heterogeneity
of iron bioavailability on plant. In this sensor, ferric iron availability to cells was assessed by
quantifying the fluorescence intensity of cells containing a plasmid-borne transcriptional
fusion between an ion-regulated promoter and gfp. Recently, Wells et al. developed an
ultrasensitive biosensor for arsenite by using laser-induced fluorescence confocal
spectroscopy to measure arsenite-stimulated enhanced green fluorescent protein synthesis
of genetically engineered E. coli bioreporter cell, which has an inherent single-molecule
detection capability.
9.2.2.2-O2-sensitive fluorescent material-based biosensor
Besides green fluorescent protein, other fluorescent materials have also been used in the
construction of microbial biosensor .Recently; fiber-optical microbial sensors for
determination of BOD were reported. The biosensors consisted of either a layer of oxygen-
sensitive fluorescent materials that are made up of seawater microorganisms immobilized in
poly (vinyl alcohol) solgel matrix and an oxygen fluorescence quenching indicator with
linear range of 4200 mg/l, or an immobilized P. putida membrane attached to an optical
fiber sensor for dissolved oxygen from ASR Co. Ltd. with detection limit of 0.5 mg/l.
9.2.3-Colorimetric biosensor
A sensitive biosensor based on color changes in the toxin sensitive colored living cells of fish
was reported. In the presence of toxins produced by microbial pathogens, the cells undergo
visible color change and the color changes in a dose-dependent manner. The results suggest
this cell-based biosensors potential application in the detection and identification of
virulence activity associated with certain air-, food-, and water-borne bacterial pathogens.
A colorimetric whole cell bioassay for the detection of common environmental pollutants
benzene, toluene, ethyl benzene and xylene (BTEX), found at underground fuel storage
tanks, using recombinant E. coli expressing toluene dioxygenase and toluene dihydrodiol
dehydrogenase was reported.
 Figure: Schematic diagram of a calorimetric biosensor. The
sample stream (a) passes through the outer insulated box (b) to the heat exchanger (c) within an
aluminium block (d). From there, it flows past the reference thermistor (e) and into the packed bed
bioreactor (f, 1ml volume), containing the biocatalyst, where the reaction occurs. The change in
temperature is determined by the thermistor (g) and the solution passed to waste (h). External
electronics (l) determines the difference in the resistance, and hence temperature, between the
thermistors.

9.3-Other types of microbial biosensors

Besides electrochemical, optical and colorimetric microbial biosensors, there are few other
types of biosensors reported recently.
9.3.1- Sensors based on baroxymeter for the detection of pressure
change
As a new application, baroxymeter has been developed as a portable wastewater direct
toxicity assessment device based on manometric bacterial respirometry. Respirometrywas
measured as the pressure drop in the headspace of a close vessel due to oxygen uptake by
the microorganism in contact with sample. This microbial pressure sensor showed good
reproducibility and comparable responses with other reported methods.
9.3.2-Sensors based on infrared analyzer for the detection of the
microbial respiration product CO2
A new method was reported for monitoring inhibitory effects in wastewater treatment plants
based on continuous measurement of the microbial respiration product CO 2. Activated
sludge microbes are used as the biological elements and their respiratory activity is inhibited
by the presence of toxic compounds, resulting in a decrease in CO 2 concentration which was
analyzed by using a CO2 infrared analyzer. Based on the measurement of CO 2 concentration
in the off gas produced during degradation of carbon compound by microbial respiration
activities, a microbial biosensor was developed to monitor the extent of organic pollution in
wastewater both off-line in a laboratory and online in a wastewater treatment plant.

10-ENVIRONMENTAL APPLICATIONS of microbial biosensors

Among microbial sensors for the analysis of toxic compounds and pollutants there is a lot of
bioluminescent sensors providing highly selective detection at a level of 10 -9 10-7 M (in some cases,
up to 10-12 M).

Toxicity
In environmental pollution monitoring, it is becoming a general opinion that chemical
analysis by itself does not provide sufficient information to assess the ecological risk of
polluted waters and wastewaters. Thus, much effort has been made during the last years to
develop and use different bioassays and biosensors for toxicity evaluation of water samples.
Whole organisms are used to measure the potential biological impact (toxicity) of a
water or soil sample. That is the case of the toxicity assays Microtox . These systems
are based on the use of luminescent bacteria, Vibrio fischeri, to measure toxicity from
environmental samples. Bacterial bioluminescence has proved to be a convenient
measure of cellular metabolism and, consequently, a reliable sensor for measuring
the presence of toxic chemicals in aquatic samples.
Some bioassay methods are integrated now in biosensors such as the Cellsense ,
which is an amperometric sensor that incorporates Escherichia coli bacterial cells for
rapid ecotoxicity analysis.
Pesticides detection
A biosensor for direct detection of organophosphorous neurotoxins has been proposed. The biosensor
is based on E. coli recombinant bacteria expressing organophosphate hydrolase and immobilized on a
pH-electrode. The developed biosensor can be used for detection of a wide range of
organophosphorous pesticides and chemical warfare agents.

Alkanes, aromatic compounds, and polycyclic aromatic

hydrocarbons (PAHs)
Contamination of soils and surface and groundwater supplies with petroleum products is a
serious environmental problem. Of particular concern for drinking water quality are water-
soluble aromatic components (e.g., benzene, toluene, ethylbenzene, and xylenes) of
petroleum products. Although many of these contaminants are readily biodegradable, they
often persist in the environment. A green fluorescent protein-based Pseudomonas
fluorescens strain biosensor was constructed and characterized for its potential to measure
benzene, toluene, ethylbenzene, and related compounds in aqueous solutions. The
biosensor is based on a plasmid carrying the toluene-benzene transcriptional activator.
Another microbial whole-cell biosensor, using E. coli with the promoter luciferase luxAB
gene, was developed for the determination of water-dissolved linear alkanes by
luminescence. The biosensor was used to detect the bioavailable concentration of alkanes in
heating oil-contaminated groundwater samples.

Determination of hydrocarbons and their derivatives

Electrochemical microbial biosensor systems for detection of a wide range of xenobiotics have
been described (Riedel et al., 1993; Riedel et al., 1991). For example, an amperomteric
biosensor for detection of monoaromatics was based on Rhodococcus cells and their extract
(Riedel et al., 1991).
The microbial biosensor for benzene detection based on an oxygen electrode and P. putida
cells was described. The sensor is supposed to be used for analyzing industrial wastewater and
groundwater (Tan et al., 1994).
The microbial biosensor for naphthalene detection based on an oxygen electrode and
Sphingomonas sp. and Pseudomonas fluorescens strains was presented in the work (Konig et
al., 1996). The lower limit of naphthalene detection was about 0.01 mg/l. The sensor
maintained its activity for 20 days.
Some of the microbial biosensor models based on the Clark-type electrode and
strainsdestructors
Of different compounds belonging to the genera Pseudomonas, Sphinomonas, Rhodococcus,
were
described in the work.
The bioluminescent bacterial sensor for detection of halogenated organic acids was based on a
transgenic E. coli strain, where the reporter luxCDABE genes from Photorhabdus
luminescens were under the control of promoter DL-2 from the strain Pseudomonas DL-DEX.
The luminescence increased by 50% in the presence of 100 mg/l of 2-chloropropionic acid. The
sensor was highly specific; measurement time was no more than 60 min.
Detection of metal ions and inorganic acids
Microbial biosensors have become widespread in the recent decade and based mainly on
genetically modified microorganisms and optical detection. Construction of microbial
bioluminescent and catalytic sensors for detection of heavy metal and inorganic acids ions is a
rather widespread approach in biosensor analysis; so, the work (Ramanathan et al., 1997) is a
survey of bacterial biosensors for detection of heavy metals. The approach is based on genetic
modification of bacteria by DNA fusion bearing the regulatory region of the metal-resistance
operon and reporter genes.
The optical bacterial biosensor for zinc and copper detection in soil samples was based on
the consortium containing the reporter bioluminescent Rhizobium leguminosarum biovar trifolii
and E. coli strains.
The microbial sensor for cyanide detection was based on S. cerevisiae cells and an oxygen
electrode. The principle of detection was registration of decrease of response to glucose in the
presence of cyanide.
Nitrate
The increasing nitrate levels found in ground and surface waters are of concern
because they can harm the water environment. In line with this, urban wastewater
treatment regulations aim to reduce pollution, including nitrate pollution, from sewage
treatment works and industry. A biosensor containing immobilized denitrifying bacteria was
applied for the determination of NO3 in tap water. Through the reduction of NO3 in a
reaction chamber, N2O was formed and determined by a N2O microelectrode, which was the
sensing element of the biosensor. A microscale biosensor for nitrate/nitrite determination
was used for in-site monitoring in an activated sludge plant. The biosensor was based on the
diffusion of nitrate/nitrite through a tip membrane into a dense mass of bacteria converting
the ions into nitrous oxide with subsequent electrochemical detection. The microbial biosensor
for nitrite detection presented in the work (Karube et al., 1982c) was based on the strain of
Nitrobacter sp. isolated from the active sludge of food plant treatment facilities.
The microbial biosensor for sulfate detection was developed on the basis of Thiobacillus
ferrooxidans and oxygen electrode. The possibility of using this sensor for sulfate detection in
rain water was shown.
Inorganic phosphate
Inorganic phosphate found in surface waters is used as a measure of the degree of
eutrophism. Traditional methods for its determination are chromatography, volumetric
titration, or spectrophotometry..The bioluminescent sensor based on recombinant cyanobacteria
Synechococcus was used for detection of bioavailable phosphorus in water reservoirs (actually, the
probability of "bloom" was assessed). The developed model was named as CyanoSensor and provided
phosphate assessment in the concentration range of 0.3 8 M.
Detection of alcohols and organic acids
A microbial sensor for ethanol detection based on G. oxydans cells and glassy carbon electrode
modified by ferricyanide was described. The selectivity of the sensor was higher over enzyme
sensors due to the application of a membrane barrier for glucose. The biosensor was employed
for real-time ethanol monitoring during periodical fermentation of glucose by immobilized
yeast.
The microbial cells possessing high selectivity are often used in biosensors for alcohol
detection along with the enzymes.
An original approach was used in the study related to development of a biosensor based on
Chlorella cells and oxygen electrode for detection of volatile compound vapors. The
characteristic feature of the approach was the registration of the oxygen produced by the cells
has been employed. Methanol was used as the model analyte. The sensor retained 50% of
initial activity after 10 days from the start of operating (Naessens & Tran-Minh, 1998a).
PCBs
Polychlorinated biphenyls (PCBs) are ubiquitous environmental pollutants widely used as
industrial chemicals, particularly as dielectric fluids in electrical transformers and capacitors.
The high toxicity of some PCB congeners represents a risk for public health as these
compounds are still present in the environment, even though the production of PCBs has
been banned in several countries many years ago. Different biosensor configurations have
been designed to determine PCBs in the environment, and these include the DNA biosensor
with chronopotentiometric detection.
Surfactants
Detergent products use surfactants as the basic active component. The anionic
surfactants are the most widely used, while the cationic surfactants represent only 5 % of
the total [70]. An amperometric biosensor for detection of anionic surfactants was
constructed with Pseudomonas rathonis T (bearing a plasmid for surfactant degradation) as
a biological element. The sensitivity and selectivity of biosensors based on bacterial strain
Pseudomonas and Achromobacter and their ability to degrade the anionic surfactants. The
degradation of surfactants by the bacteria caused a decrease in dissolved oxygen and a
change in the oxygen electrode current.
11-Future trends
Since Clark and Lyons developed the first biosensor, the field of biosensors has greatly
expanded. Microorganisms, due to their low cost, long lifetime and wide range of suitable pH
and temperature, have been widely employed as the biosensing element in the construction
of biosensors [2]. Even though a variety of microbial biosensors have been developed for
environmental, food, military and biomedical application, when compared to enzyme
biosensors the development of highly satisfactory microbial biosensor is still hampered
because they suffer from
long response time, low sensitivity and poor selectivity.
With a better understanding of the genetic information of microbes and the development of
improved recombinant
DNA technologies, different enzymes and proteins have been expressed on the cell surface
through surface expression anchors. In this format, the microbes can serve as an enzymes
support matrix. The surface expressed enzymes or proteins can directly react with
substrates without the entry of substrates into the microbes. Through thisway, faster
response and highly sensitive microbial biosensors can be developed.