Biosensing of arsenic by whole-cell bacterial bioreporter immobilized on polycaprolactone (PCL) electrospun fiber.

In recent years, heavy metals derived from several anthropogenic sources have both direct and indirect detrimental effects on the health of the environment and living organisms. Whole-cell bioreporters (WCBs) that can be used to monitor the levels of heavy metals in drinking and natural spring waters are important. In this study, whole-cell arsenic bacterial bioreporters were immobilized using polycaprolactone (PCL) electrospun fibers as the support material. The aim is to determine the properties of this immobilized bioreporter system by evaluating its performance in arsenic detection. Within the scope of the study, different growth media and fiber immobilization times were tested to determine the parameters affecting the fluorescent signals emitted by the immobilized bioreporter system in the presence of two dominant forms of arsenic, namely arsenite (As(III)) and arsenate (As(V)). In addition, the sensitivity, selectivity, response time, and shelf-life of the developed bioreporter system were evaluated. As far as the literature is concerned, this is the first study to investigate the potential of using PCL-electrospun fiber-immobilized fluorescent bacterial bioreporter for arsenic detection. This study will open new avenues in environmental arsenic monitoring.

Optimization of whole-cell bacterial bioreporter immobilization on electrospun cellulose acetate (CA) and polycaprolactone (PCL) fibers for arsenic detection.

Arsenic contamination is a critical global problem, and its widespread environmental detection is becoming a prominent issue. Herein, electrospun fibers of cellulose acetate (CA) and polycaprolactone (PCL) were successfully fabricated and used as the support material for immobilization of arsenic-sensing bacterial bioreporter for the first time. To date, no attempt has been made to immobilize fluorescent whole-cell bioreporter cells on electrospun fibers for arsenic detection. CA and PCL electrospun fibers were fabricated via traditional electrospinning technique and characterized by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), and contact angle meter. Following immobilization of the bacterial bioreporter cells, the immobilized bacteria were also characterized by viability assay using AlamarBlue. The effects of growth phase and cell concentration on the fluorescence response of fiber-immobilized arsenic bioreporters to arsenic were also investigated. After immobilization of arsenic bioreporters on 10 wt% PCL fiber, 91% of bacterial cells remained viable, while this value was 55.4% for cells immobilized on 12.5 wt% CA fiber. Bioreporter cells in the exponential growth phase were shown to be more sensitive to arsenic compared to aged cells. While both the electropsun PCL- and CA-immobilized bioreporters successfully detected 50 and 100 µg/L of arsenite (As (III)) concentrations, the PCL-immobilized bioreporter showed better fluorescence performance which should be investigated in future studies. This study helps to fill some gaps in the literature and demonstrates the potential for using electrospun fiber-immobilized arsenic whole-cell bioreporter for arsenic detection in water.

Use of whole-cell bioreporters to assess bioavailability of contaminants in aquatic systems.

Water contamination has become increasingly a critical global environmental issue that threatens human and ecosystems' health. Monitoring and risk assessment of toxic pollutants in water bodies is essential to identifying water pollution treatment needs. Compared with the traditional monitoring approaches, environmental biosensing via whole-cell bioreporters (WCBs) has exhibited excellent capabilities for detecting bioavailability of multiple pollutants by providing a fast, simple, versatile and economical way for environmental risk assessment. The performance of WCBs is determined by its elements of construction, such as host strain, regulatory and reporter genes, as well as experimental conditions. Previously, numerous studies have focused on the design and construction of WCB rather than improving the detection process and commercialization of this technology. For investigators working in the environmental field, WCB can be used to detect pollutants is more important than how they are constructed. This work provides a review of the development of WCBs and a brief introduction to genetic construction strategies and aims to summarize key studies on the application of WCB technology in detection of water contaminants, including organic pollutants and heavy metals. In addition, the current status of commercialization of WCBs is highlighted.

Immobilization of fluorescent bacterial bioreporter for arsenic detection.

Whole-cell bacterial biosensors hold great promise as a practical complementary approach for in-field detection of arsenic. Although there are various bacterial bioreporter systems for arsenic detection, fewer studies reported the immobilization of arsenic bioreporters. This study aimed at determining immobilization of specific bacterial bioreporter in agar and alginate biopolymers to measure level of arsenite and/or arsenate. To achieve sensitive detection, immobilization parameters of polymer concentration and cell density were evaluated. Moreover, by changing the culture medium, immobilized bioreporter cells in minimal medium can detect arsenite while they can detect both arsenite and arsenate in phosphate-limited minimal medium. When optimal parameters were applied, agar and alginate immobilized bioreporter systems can detect arsenite and arsenate concentrations of 10 µg/l and 200 µg/l within 5 h and 2 h, respectively. The results showed that the immobilized bacterial bioreporter systems are able to determine the concentrations of the two abundant species of arsenic; arsenite and arsenate, as opposed to other studies which reported only arsenite detection. This is the first study describe agar hydrogel and alginate bead immobilization of fluorescent arsenic bacterial bioreporter that can detect both arsenite and arsenate at the safe drinking water limit. Thus, this study will enable further steps to be taken towards developing sensitive and selective portable devices to assess environmental arsenic contamination and prevent acute arsenic toxicity.

Evaluation of novel thermo-resistant Micractinium and Scenedesmus sp. for efficient biomass and lipid production under different temperature and nutrient regimes.

Despite the vast interest in microalgae as feedstock for biodiesel production, relatively few studies examined their response to diurnal temperature fluctuation. Here, we describe biomass and lipid productivities and fatty acid profiles of thermo-resistant Micractinium sp. and Scenedesmus sp. grown in batch cultures in a laboratory set-up that mimics a typically warm summer day in Central Anatolia with a 16-h light temperature of 30°C and 8-h dark temperature of 16°C (30°C (day)/16°C (night)). Both strains can survive a temperature range of 10-50°C. We found the lipid productivities of Micractinium sp. and Scenedesmus sp. as 30/21mgL(-1)d(-1) and 6/7mgL(-1)d(-1), respectively during the 30°C (day)/16°C (night) cycle. Saturated fatty acid content increased with increasing temperature. Additionally, we cultured Micractinium sp. under Nitrogen (N) and Phosphorus (P) limiting conditions. Highest lipid productivity of 85.4±2mgL(-1)d(-1) was obtained under P-depletion during exponential growth phase. Oleic acid amount also increased eight fold during P-deplete.