Expression of Rhizobium tropici phytochelatin synthase in Escherichia coli resulted in increased bacterial selenium nanoparticle synthesis

Yuan, Qunying; Bomma, Manjula; Hill, H.J.

doi:10.1007/s11051-020-05095-z

Cited by 5 publications

(15 citation statements)

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“…In principle, employing the soluble intracellular extracts as a reaction medium for nanoparticle synthesis can avoid the tedious and nearly impossible isolation of nanoparticles from cellular debris. In our previous study, the recombinant E. coli DH5a cells that express R. tropici phytochelatin synthase showed increased efficiency in selenium nanoparticle synthesis [16]. We postulated that the enhanced capacity of these recombinant bacteria for nanoparticle synthesis was resulted from the increased phytochelatin production upon the overexpression of R. tropici phytochelatin synthase.…”

Section: Introductionmentioning

confidence: 94%

“…The assembly of the R. tropici phytochelatin synthase gene was carried out as described by D. Marsc and Q. Yuan [16,34,35]. Glycerol stocks of E. coli DH5α cells transformed with plasmid containing the sequence-verified phytochelatin synthase gene were prepared for protein expression evaluation and nanoparticle synthesis as previously described [16]. All chemicals and reagents were purchased from Fisher Scientific (Pittsburgh, PA, USA) unless specified.…”

Section: Determination Of R Tropici Phytochelatin Synthase Protein Expression In Recombinant Ementioning

confidence: 99%

“…Preparation of E. coli DH5α cells for determining protein expression was carried out as described in reference [16]. Cell pellets were resuspended in lysis buffer containing 50 mM Tris-HCl (pH 8), 50 mM NaCl and sonicated to break apart the cells.…”

Section: Determination Of R Tropici Phytochelatin Synthase Protein Expression In Recombinant Ementioning

confidence: 99%

“…The whole lysate was centrifuge at 15,000× g for 30 min to separate the soluble proteins from the cell membrane debris. Level of protein expression was examined as described in reference [16]. Briefly, 10µL of the supernatant was loaded to 10% SDS-PAGE gel and proteins were separated by electrophoresis.…”

Section: Determination Of R Tropici Phytochelatin Synthase Protein Expression In Recombinant Ementioning

confidence: 99%

“…The contribution of phytochelatins to nanoparticle synthesis has been investigated [13][14][15]. We also developed a recombinant E. coli bacterium that expresses R. tropici phytochelatin synthase, and discovered that overexpression of R. tropici phytochelatin synthase in E. coli cells improved the bacterial heavy metal resistance, and increased yield of selenium nanoparticles when whole cells were used to synthesize nanoparticles [16]. Nevertheless, one challenge associated with whole cell nanoparticle synthesis in our previous studies was the isolation of nanoparticles from cellular debris to obtain pure nanoparticles.…”

Section: Introductionmentioning

confidence: 99%

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Enhanced Extracellular Synthesis of Gold Nanoparticles by Soluble Extracts from Escherichia coli Transformed with Rhizobium tropici Phytochelatin Synthase Gene

Yuan

Bomma

2021

Metals

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Phytochelatins, the enzymatic products of phytochelatin synthase, play a principal role in protecting the plants from heavy metal and metalloid toxicity due to their ability to scavenge metal ions. In the present study, we investigated the capacity of soluble intracellular extracts from E. coli cells expressing R. tropici phytochelatin synthase to synthesize gold nanoparticle. We discovered that the reaction mediated by soluble extracts from the recombinant E. coli cells had a higher yield of gold nanoparticles, compared to that from the control cells. The compositional and morphological properties of the gold nanoparticles synthesized by the intracellular extracts from recombinant cells and control cells were similar. In addition, this extracellular nanoparticle synthesis method produced purer gold nanoparticles, avoiding the isolation of nanoparticles from cellular debris when whole cells are used to synthesize nanoparticles. Our results suggested that phytochelatins can improve the efficiency of gold nanoparticle synthesis mediated by bacterial soluble intracellular extracts, and the potential of extracellular nanoparticle synthesis platform for the production of nanoparticles in large quantity and pure form is worth further investigation.

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Section: Introductionmentioning

confidence: 94%

Section: Determination Of R Tropici Phytochelatin Synthase Protein Expression In Recombinant Ementioning

confidence: 99%

Section: Determination Of R Tropici Phytochelatin Synthase Protein Expression In Recombinant Ementioning

confidence: 99%

Section: Determination Of R Tropici Phytochelatin Synthase Protein Expression In Recombinant Ementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Enhanced Extracellular Synthesis of Gold Nanoparticles by Soluble Extracts from Escherichia coli Transformed with Rhizobium tropici Phytochelatin Synthase Gene

Yuan

Bomma

2021

Metals

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Analysis of metal accumulation and Phytochelatin Synthase (PCS) gene expression in Saccharum spontaneum L. as regards its function for remediation of lead (Pb) contamination

Rini

Hidayati

2021

IOP Conf. Ser.: Earth Environ. Sci.

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Pollution caused by heavy metals, has become a serious problem. Adverse effects arising from the increased use of heavy metals in a variety of human activities lead to any environmental degradation. Lead (Pb) is one of most common contaminants in the environment and highly toxic. Pb is less mobile, so its compound tends to accumulate in soil and sediments. Definitely, efforts are needed to remove this contaminant in the environment. Saccharum spontaneum L. is a perennial grass which has potential to be used as an accumulator plant to clean up pollutants. The ability of this plant as metal accumulator was tested in this study. S.pontaneum plants were treated using Pb in the concentrations of 0 ppm (control), 100 ppm, 200 ppm, and 300 ppm for 8 weeks. The results showed that there was an increase in the percentage of relative accumulation of Pb in the treated plants. This also indicated that plant roots accumulated more Pb than shoots. Meanwhile, expression of Phytochelatin synthase (PCS) gene increased 1.3-to-3.5-fold inductions in roots by increasing concentration of Pb treatments for 24 h. PCS gene expression showed the higher induction in the roots than in the shoots of S.spontaneum plant under Pb treatments.

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Exploiting Rhizobium for Cadmium Sulphide Nanoparticle Synthesis: Heterologous Expression of an Escherichia coli DH10B Enzyme, YbdK [EC: 6.3.2.2] in Sinorhizobium fredii NGR234

Dave¹,

Khanna²,

Robin³

2022

J. Pure Appl. Microbiol.

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Escherichia coli DH10B has 1.1 kb ybdK gene which is responsible for encoding YbdK enzyme that possess a Gamma glutamyl cysteine synthetase activity. ybdK gene was ligated downstream of a constitutive derepressed lac promoter of a low copy number plasmid vector pBBR1MCS-2, giving rise to a recombinant plasmid pPAT. Sinorhizobium fredii NGR234 transformed with pPAT showed an augmented production of glutathione which in turn increased the production of cadmium sulphide nanoparticles to some extent. Also, a heterologous expression of YbdK in Sinorhizobium fredii NGR234 improved the oxidation status of bacterial cells which is confirmed by fluorescence microscopy images and fluorometry. Genetically modified (GM) cells stained by DCFDA showed a significant decrease in fluorescence compared to wild type (WT) cells. Physical and chemical properties of the nanoparticles produced by the pPAT transformed Sinorhizobium fredii NGR234 differed significantly compared to wild type (WT) Sinorhizobium fredii NGR234. Comparative analysis of the nanoparticles by FTIR and SEM analysis revealed the functional groups attached to nanoparticles and average nanoparticle size respectively. Nanoparticles synthesized by genetically modified (GM) bacteria were about 3 times smaller in size compared to those produced by wild type (WT) rhizobium. FTIR analysis revealed an augmented presence of peptide with the nanoparticles produced by GM bacteria compared to those produced by the WT bacteria. XRD data revealed that biosynthesized CdS nanoparticles are face centered crystalline particles which was confirmed by comparing the peaks to standard JCPDS data (JCPDS card no. 10-454).

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Expression of Rhizobium tropici phytochelatin synthase in Escherichia coli resulted in increased bacterial selenium nanoparticle synthesis

Cited by 5 publications

References 35 publications

Enhanced Extracellular Synthesis of Gold Nanoparticles by Soluble Extracts from Escherichia coli Transformed with Rhizobium tropici Phytochelatin Synthase Gene

Enhanced Extracellular Synthesis of Gold Nanoparticles by Soluble Extracts from Escherichia coli Transformed with Rhizobium tropici Phytochelatin Synthase Gene

Analysis of metal accumulation and Phytochelatin Synthase (PCS) gene expression in Saccharum spontaneum L. as regards its function for remediation of lead (Pb) contamination

Exploiting Rhizobium for Cadmium Sulphide Nanoparticle Synthesis: Heterologous Expression of an Escherichia coli DH10B Enzyme, YbdK [EC: 6.3.2.2] in Sinorhizobium fredii NGR234

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