A novel bacterial isolate Stenotrophomonas maltophilia as living factory for synthesis of gold nanoparticles

<p>Abstract</p> <p>Background</p> <p>The synthesis of gold nanoparticles (GNPs) has received considerable attention with their potential applications in various life sciences related applications. Recently, there has been tremendous excitement in the study of nanoparticles synthesis by using some natural biological system, which has led to the development of various biomimetic approaches for the growth of advanced nanomaterials. In the present study, we have demonstrated the synthesis of gold nanoparticles by a novel bacterial strain isolated from a site near the famous gold mines in India. A promising mechanism for the biosynthesis of GNPs by this strain and their stabilization via charge capping was investigated.</p> <p>Results</p> <p>A bacterial isolate capable of gold nanoparticle synthesis was isolated and identified as a novel strain of <it>Stenotrophomonas malophilia </it>(AuRed02) based on its morphology and an analysis of its 16S rDNA gene sequence. After 8 hrs of incubation, monodisperse preparation of gold nanoparticles was obtained. Gold nanoparticles were characterized and found to be of ~40 nm size. Electrophoresis, Zeta potential and FTIR measurements confirmed that the particles are capped with negatively charged phosphate groups from NADP rendering them stable in aqueous medium.</p> <p>Conclusion</p> <p>The process of synthesis of well-dispersed nanoparticles using a novel microorganism isolated from the gold enriched soil sample has been reported in this study, leading to the development of an easy bioprocess for synthesis of GNPs. This is the first study in which an extensive characterization of the indigenous bacterium isolated from the actual gold enriched soil was conducted. Promising mechanism for the biosynthesis of GNPs by the strain and their stabilization via charge capping is suggested, which involves an NADPH-dependent reductase enzyme that reduces Au³⁺to Au⁰through electron shuttle enzymatic metal reduction process.</p

Nations within a nation: variations in epidemiological transition across the states of India, 1990–2016 in the Global Burden of Disease Study

18% of the world's population lives in India, and many states of India have populations similar to those of large countries. Action to effectively improve population health in India requires availability of reliable and comprehensive state-level estimates of disease burden and risk factors over time. Such comprehensive estimates have not been available so far for all major diseases and risk factors. Thus, we aimed to estimate the disease burden and risk factors in every state of India as part of the Global Burden of Disease (GBD) Study 2016

Quartz Crystal Based Microgravimetric Immunobiosensors

Piezoelectric crystal immunosensors have attracted considerable interest in the last few years since the monitoring of a specific substance is central in many applications. These sensors work on the principle of measuring a small change in resonant frequency of an oscillating piezoelectric crystal due to change in mass on the sensor surface. Because of their low cost and high Q-factors, these miniaturized sensors show fast response time, high sensitivity, and are suitable for mass production using standard fabrication techniques. Arrays of such sensors could be fabricated to cover ranges of a particular sensing property and have the potential for seamless integration with CMOS-based electronics. The present article demonstrates a type of piezoelectric crystal immunosensor using simple design of frequency oscillator and monitoring circuitry for biomedical applications. The basic criterion for immobilizing biomolecules (receptor etc.) on sensor surface for immunoassay application is briefly discussed

Activating piezoelectric crystal surface by silanization for microgravimetric immunobiosensor application

The development of a microgravimetric immunobiosensor using a piezoelectric quartz crystal as a detector requires a stable and reproducible immobilization method for ligand binding. The method of silanization using 3-aminopropyltriethoxysilane (APTES) has been widely used for activating the carrier surface. In the present study, APTES deposition on a piezoelectric crystal surface was studied under various solvent conditions. A fluorescence method, using fluorescence isothiocyanate as a dye, was demonstrated for the quantification of amino groups on the silanized piezoelectric crystal surface. The optimum binding conditions of APTES deposition on a piezoelectric crystal surface were incorporated for the covalent immobilization of protein on the crystal surface in developing a stable and sensitive microgravimetric immunobiosensor. Determination of immunoglobulin G (IgG) concentration was performed using APTES modified piezoelectric crystals coated with protein G. The resonant frequency shift, resulting from the formation of protein G-IgG complex on the crystal surface, correlated with the concentration of IgG in the range 10 ng/ml to 0.1 mg/ml. The APTES modified, protein G coated crystals were found to be quite stable and did not show a significant loss of sensitivity even after 12 weeks of storage at 4°C in a desiccator

Determination of immunoglobulin M concentration by piezoelectric crystal immunobiosensor coated with protamine

In the present study, the specific binding between protamine and immunoglobulin M (IgM) has been exploited to construct a piezoelectric crystal based immunobiosensor for the determination of concentration of IgM. The system consisted of highly stable IC based oscillator, 8-digit frequency counter and modified piezoelectric crystal device. The crystal surface was physically modified and chemically treated (refluxed) with strong acid to produce stable hydroxylic groups of silicon oxide. This modified surface reacted strongly with coupling reagents for binding of protein molecules. The protamine was immobilized by using either γ-aminopropyltriethoxy silane (γ-APTES) or 2.2.2-trifluoroethanesulfonyl chloride (tresyl chloride). Scanning electron microscope images of piezo crystal revealed that tresyl activated surface presented more surface area for binding than γ-APTES modified surface and showed better sensitivity. This immobilization technique also improved the reproducibility and long term stability of the detection system. Using the system described, the IgM concentration up to the level of 10 ng/ml could be detected without interference of IgG

Characterization of Hapten–Protein Conjugates: Antibody Generation and Immunoassay Development for Pesticides Monitoring

The generation of specific and sensitive antibodies against small molecules is greatly dependent upon the characteristics of the hapten–protein conjugates. In the present study, we report a new fluorescence-based method for the characterization of hapten–protein conjugates. The method is based on an effect promoted by hapten–protein conjugation density upon the fluorescence intensity of the intrinsic tryptophan chromophore molecules of the protein. The proposed methodology is applied to quantify the hapten–protein conjugation density of two different class of pesticides (atrazine and 2,4-dichlorophenoxyacetic acid in this study) coupled to carrier protein. The study proved useful for monitoring the course of hapten–protein conjugation for the production of specific antibodies against small molecules. Well-characterized hapten–protein conjugates enabled obtaining highly sensitive anti-atrazine and anti-2,4-D antibodies with IC(50) values equal to 12 and 70 ng mL(−1) for atrazine and 2,4-D respectively. These antibodies were used for developing a fluorescence-based immunoassays format demonstrating a detection limit of atrazine and 2,4-D in standard water samples 2 and 7 ng mL(−1), respectively. The developed immunoassay format could be used as convenient quantitative tools for sensitive and specific screening of pesticides in samples

Development of piezoelectric crystal based microgravimetric immunoassay for determination of insulin concentration

A microgravimetric, piezoelectric crystal based immunoassay for the quantification of insulin concentration is described. The method utilizes a modified piezoelectric crystal device having an antibody specific to insulin bound to its surface. The antibody to insulin was immobilized on the surface of crystal electrode by using either 3-aminopropyltriethoxy silane (3-APTES), polyethyleneimine (PEI) or covalently coupled protein A-gold immobilization method. Coating an electrode with a cross linked protein A-antibody complex gave better results in terms of sensitivity and stability. Using the system described, the insulin concentration up to 1 ng ml- could be detected. The stability and reusability of the system was further improved by using a mild eluting reagent which successfully removed the bound insulin molecules from the antibody-coated crystal without affecting the immobilized insulin antibody. Scanning tunneling microscopic (STM) study was also done to confirm the surface coverage and orientation of insulin and antibody molecules on the modified piezoelectric crystal electrode surface. A comparison between the present study and the well-established radioimmunoassay technique (RIA) revealed that the described microgravimetric immunoassay technique (MIA) could successfully be developed as an alternative of RIA