Late state‐of‐the‐art analytical methodologies in chromatography, spectroscopy, and electroanalysis have been developed to meet the challenges of changing environmental and health issues. The modern trends in developing new protocols emphasize economic, portable, nano, or even smaller sample sizes and less time‐consuming processes. This has led to the development of technology‐based biosensors which meet most of the above requirements. The lab‐on‐chip technology exploiting enzyme‐based biosensors has made the analytical processes very efficient, accurate, affordable, and requiring nano‐scale sample sizes. In this review, an attempt is being made to review the literature based on state‐of‐the‐art technology of enzyme‐based biosensors for the detection of biomolecules.
Efficient healthcare management demands prompt decision-making based on fast diagnostics tools, astute data analysis, and informatics analysis. The rapid detection of analytes at the point of care is ensured using microfluidics in synergy with nanotechnology and biotechnology. The nanobiosensors use nanotechnology for testing, rapid disease diagnosis, monitoring, and management. In essence, nanobiosensors detect biomolecules through bioreceptors by modulating the physicochemical signals generating an optical and electrical signal as an outcome of the binding of a biomolecule with the help of a transducer. The nanobiosensors are sensitive and selective and play a significant role in the early identification of diseases. This article reviews the detection method used with the microfluidics platform for nanobiosensors and illustrates the benefits of combining microfluidics and nanobiosensing techniques by various examples. The fundamental aspects, and their application are discussed to illustrate the advancement in the development of microfluidics-based nanobiosensors and the current trends of these nano-sized sensors for point-of-care diagnosis of various diseases and their function in healthcare monitoring.
Screening of 20,000 clones of a fosmid gene bank, constructed from DNA extracted from North West Himalaya (NWH) glacier soil sample, using functional approach identified 10 esterase/lipase-producing clones. Of these, a clone designated pFG43 with an insert size of 45 kb which produced the highest concentration of enzyme (467.43 U/mg) was sequenced. Clone pFG43 contained 61 open reading frames (ORF) and of these an ORF of 1155 bp designated ME-003, was found to be closely related to a hydrolase from Acidobacteria sps (77% sequence identity and E value = 1e-164) and subsequently identified as a putative cocaine esterase. ORF ME-003 was amplified and sub-cloned using a TA vector system into E. coli (DH5α). The purified recombinant enzyme with a molecular weight of 43 kDa had optimal activity at 40 °C, pH 6 and the highest activity with shorter chain fatty acids than with higher chain length fatty acids. There is insignificant effect of inhibitors on the enzyme activity of ME-003, except PMSF which completely inhibited its activity. ME-003 activity was also inhibited in the presence of copper oxide but remained stable in presence of other metal ions. The enzyme activity was also inhibited in the presence of organic solvents; however, in the presence of 10% isopropanol, 12% of enzymatic activity was retained. Among various detergents, SDS completely inhibited enzymatic activity. The recombinant enzyme also shows enantio-specific activity against the racemic drug intermediates/precursors and exhibited 90% ee against racemic 1-phenyl ethanol and fluoxetine.
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