Isolation and characterization of a recombinant class C acid phosphatase from Sphingobium sp. RSMS strain

Rangu, Shyam Sunder; Singh, Rahul; Gaur, Neeraj; Rath, Devashish; Makde, Ravindra D.; Mukhopadhyaya, Robin

doi:10.1016/j.btre.2022.e00709

Cited by 4 publications

(5 citation statements)

References 31 publications

(42 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It is best characterized in Sphingomonas sp. RSMS (25,26), which is clearly distinct from the incomplete pathway characterization for Rhodopseudomonas palustris (24). As a result, the phosphostriesterase genes we observed as potentially involved with tributyl phosphate degradation may represent a novel pathway.…”

Section: Discussionmentioning

confidence: 71%

“…We found no homology between any genes in our biomarker MAGs and CYP201A2 from R. palustris (24) or aps from Sphingomonas sp. RSMS (25). Therefore, we focused on phosphoesterase genes which may be involved in degrading tributyl phosphate to dibutyl phosphate (phosphotriesterases), dibutyl phosphate to monobutyl phosphate (phosphodiesterases) and monobutyl phosphate to butanol and inorganic phosphate (phosphomonoesterase or acid phosphatase) (25,26).…”

Section: Phosphotriesterase Genes Found In Biomarker Magsmentioning

confidence: 99%

“…RSMS (25). Therefore, we focused on phosphoesterase genes which may be involved in degrading tributyl phosphate to dibutyl phosphate (phosphotriesterases), dibutyl phosphate to monobutyl phosphate (phosphodiesterases) and monobutyl phosphate to butanol and inorganic phosphate (phosphomonoesterase or acid phosphatase) (25,26). Potential phosphtriesterases were identified, and their distribution across MAGs supports their role in promoting growth in response to tributyl phosphate.…”

Section: Phosphotriesterase Genes Found In Biomarker Magsmentioning

confidence: 99%

See 2 more Smart Citations

Response of soil bacteria to PUREX chemicals suggests biomarker utility and bioremediation potential

Podowski,

Forrester,

Antonopoulos

et al. 2024

J Radioanal Nucl Chem

View full text Add to dashboard Cite

Chemicals involved in plutonium uranium reduction extraction (PUREX) have the potential to be released from nuclear reprocessing facilities and accumulate in the environment. In order to understand how soil microbial communities respond to contamination by PUREX chemicals, we carried out a series of microcosm experiments, exposing chemically diverse soils to a range of concentrations of key chemicals used in the PUREX process. We tested 4 PUREX chemicals, and 5 soil types using 16S rRNA amplicon sequencing, determining that responses of microbial communities are dependent on the soil type in which they reside, and that tributyl phosphate exposure appears to generate the most reproducible and detectable shifts in microbial communities. We identified a number of key taxa that are consistently enriched in soils exposed to tributyl phosphate. These key taxa are either in the family Rhizobiaceae or genus Pseudomonas. The relative abundance of these key taxa is concentration dependent, and their abundance remains elevated at least 100 days post initial exposure. Using whole-shotgun metagenomic sequencing, we reconstructed the genomes of these key taxa and find a number of putative phosphotriesterase genes found only in Rhizobiaceae. We find the abundance of phosphotriesterase genes is significantly higher in samples exposed to tributyl phosphate. These phosphotriesterase genes, which degrade tributyl phosphate into dibutyl phosphate and butanol, may serve as effective biomarkers for tributyl phosphate contaminated soil, as well as a method for future bioremediation.Importance: Nuclear materials reprocessing facilities have the capacity to release toxic chemicals during normal operations or accidents. This study examines the ways in which chemicals involved with nuclear materials reprocessing impact microorganisms in the soil. Our intention was to understand the consequences of the release of these chemicals on ecosystems that may surround these reprocessing facilities. We find soil microbial communities change in response to some chemicals but not others, and that tributyl phosphate appears to generate the most reproducible and detectable shifts in microbial communities. Microorganisms in the family Rhizobiaceae increase in abundance in response to the addition of tributyl phosphate, and an examination of the genomes of these microbes suggest they may be able to break down tributyl phosphate to access the phosphosphate present in this chemical. Overall, this work demonstrates that changes in soil microbial communities in response to contamination with chemicals from nuclear materials reprocessing facilities may be predictable, and these responses could be leveraged to remediate contamination.

show abstract

Section: Discussionmentioning

confidence: 71%

Section: Phosphotriesterase Genes Found In Biomarker Magsmentioning

confidence: 99%

Section: Phosphotriesterase Genes Found In Biomarker Magsmentioning

confidence: 99%

See 1 more Smart Citation

Response of soil bacteria to PUREX chemicals suggests biomarker utility and bioremediation potential

Podowski,

Forrester,

Antonopoulos

et al. 2024

J Radioanal Nucl Chem

View full text Add to dashboard Cite

show abstract

“…It should be noted that the Thermus enzyme displayed optimal temperature at 70°C. Currently, our efforts are focused on crystallizing FS6, as the apparent oligomeric state suggests a possible decameric structure, in contrast to the dimeric form observed in previously crystallized proteins (Rangu et al, 2022;Felts et al, 2007;Singh et al, 2011 andFelts et al, 2006). Although the exact nature of subunit interactions remains unknown, it is worth mentioning that the rPmCCAP protein of Pasteurella multocida has been described as a trimer of dimers (Singh et al, 2011).…”

Section: Discussionmentioning

confidence: 99%

“…Overall, it seems that the FS6 enzyme shows pH‐independent Km values with PNPP, contrasting with the pH‐dependent behaviour observed for the Clostridium enzyme. Additionally, the FS6 enzyme showed greater efficiency with physiological substrates compared to PNPP, which differs from the Clostridium and Sphingobium enzyme's performance (Reilly et al., 2009 ; Rangu et al., 2022 ). While FS6 exhibits low identity with the Thermus thermophilus extremozyme, it functions effectively across a wide range pH and high temperatures.…”

Section: Discussionmentioning

confidence: 99%

Characterization of an extremophile bacterial acid phosphatase derived from metagenomics analysis

Recio,

de la Torre,

Daddaoua

et al. 2024

Microbial Biotechnology

View full text Add to dashboard Cite

Acid phosphatases are enzymes that play a crucial role in the hydrolysis of various organophosphorous molecules. A putative acid phosphatase called FS6 was identified using genetic profiles and sequences from different environments. FS6 showed high sequence similarity to type C acid phosphatases and retained more than 30% of consensus residues in its protein sequence. A histidine‐tagged recombinant FS6 produced in Escherichia coli exhibited extremophile properties, functioning effectively in a broad pH range between 3.5 and 8.5. The enzyme demonstrated optimal activity at temperatures between 25 and 50°C, with a melting temperature of 51.6°C. Kinetic parameters were determined using various substrates, and the reaction catalysed by FS6 with physiological substrates was at least 100‐fold more efficient than with p‐nitrophenyl phosphate. Furthermore, FS6 was found to be a decamer in solution, unlike the dimeric forms of crystallized proteins in its family.

show abstract

Methods to Characterise Enzyme Kinetics with Biological and Medicinal Substrates: The Case of Alkaline Phosphatase

Harroun

Vallée‐Bélisle

2023

Chemistry Methods

View full text Add to dashboard Cite

Alkaline phosphatase (AP) enzymes are of broad interest in fields ranging from biochemistry and medicine to biotechnology and nanotechnology. Characterising the catalytic activity of AP is typically realised by either employing non‐natural signal‐generating substrates that are detectable by absorbance and fluorescence spectroscopy or by quantifying the release of inorganic phosphate by the classic malachite green assay. The latter method is often required for studying “spectroscopically silent” biomolecular substrates, but it does not enable continuous monitoring of kinetics in real‐time. In recent years, newer techniques for studying AP function have been developed to circumvent this limitation, including fluorescent and colourimetric substrate‐specific assays based on supramolecular chemistry, organic probes and nanomaterials, as well as other assays based on isothermal titration calorimetry, direct detection with infrared spectroscopy and mass spectrometry, and monitoring conformational change by fluorescent nanoantennas. Here, we review these strategies and comment on their strengths and weaknesses in the context of AP.

show abstract

Isolation and characterization of a recombinant class C acid phosphatase from Sphingobium sp. RSMS strain

Cited by 4 publications

References 31 publications

Response of soil bacteria to PUREX chemicals suggests biomarker utility and bioremediation potential

Response of soil bacteria to PUREX chemicals suggests biomarker utility and bioremediation potential

Characterization of an extremophile bacterial acid phosphatase derived from metagenomics analysis

Methods to Characterise Enzyme Kinetics with Biological and Medicinal Substrates: The Case of Alkaline Phosphatase

Contact Info

Product

Resources

About