Expression and purification of an ArsM-elastin-like polypeptide fusion and its enzymatic properties

Ke, Changdong; Xiong, Hui; Zhao, Chungui; Zhang, Zhigang; Zhao, Xiao-Lan; Rensing, Christopher; Zhang, Guangya; Yang, Suping

doi:10.1007/s00253-019-09638-w

Cited by 8 publications

(4 citation statements)

References 42 publications

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“…Previous studies had shown that ArsM encoded on the genome of R. palustris CGA009 displayed a high methylation acitivity [34]; therefore, a series of studies have been conducted with this enzyme, which is considered as a suitable candidate for an applied As methylation process in bioremediation [17,34,50,51]. However, this study and previous studies [26] showed that R. palustris CGA009 did not exhibit a significant As volatilization rate with the rate reaching less than 1% of the total As (25.0 µmol/L As(III)).…”

Section: Discussionmentioning

confidence: 96%

“…In view of the fact that bioremediation could better maintain soil structure and ecological balance, bioremediation of As by microorganisms has been advocated because of the microorganism's potential advantages in facilitating economically viable and environmental friendly technologies [36][37][38]. The final product of As methylation, gaseous trimethylarsine (TMAs(III)), could remove As from contaminated sites by As volatilization, thereby increasing the possibilities for bioremediation of As-contaminated environment [8,10,34]. As methylation is widespread in natural environment, and the genes encoding ArsM were shown to be distributed widely in various environments (e.g., paddy soil, groundwater, acid mine drainage and composting manure) [22,[39][40][41][42] and in every kingdoms of life [43].…”

Section: Discussionmentioning

confidence: 99%

“…As efflux transporters reduce intracellular As concentrations by pumping out the corresponding As compounds, thus potentially limiting microbial As methylation and subsequent As volatilization. Previous studies have shown that deletion or inactivation of genes encoding As efflux transporters increased intracellular As accumulation [ 33]; Arsenicibacter rosenii SM-1 exhibited a higher As volatilization (47.6±18.4%) due to the lack of genes encoding As efflux transporters on its genome [9], and engineered E. coli strain lacking a gene encoding an As efflux pump displayed a higher As volatilization rate (~10.39%) [34]. Therefore, As efflux transporters limit the microbial As methylation rate.…”

Section: However Little Information Is Available About the Genetic Dmentioning

confidence: 99%

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Arsenite efflux limits microbial arsenic volatilization

Ke¹,

Kuang²,

Zhao³

et al. 2019

Preprint

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Background: Arsenic (As) methylation is regarded as a potential way to volatize and thereby remove As from the environment. However, most microorganisms conducting As methylation display low As volatilization efficiency as As methylation is limited by As efflux transporters as both processes compete for arsenite [As(III)]. In the study, we deleted arsB and acr3 from Rhodopseudomonas palustris CGA009, a good model organism for studying As detoxification, and further investigated the effect of As(III) efflux transporters on As methylation. Results: Two mutants were obtained by gene deletion. Compared to the growth inhibition rate (IC50) [1.57±0.11 mmol/L As(III) and 2.67±0.04 mmol/L arsenate [As(V)] of wildtype R. palustris CGA009, the As(III) and As(V) resistance of the mutants decreased, and IC50 value of the R. palustris CGA009 ∆arsB mutant was 1.47±0.02 mmol/L As(III) and 2.12±0.03 mmol/L As(V), respectively, and that of the R. palustris CGA009 ∆acr3 mutant was 1.21±0.07 mmol/L As(III) and 1.76±0.12 mmol/L As(V), respectively. The As volatilization rate of R. palustris CGA009 ∆arsB and R. palustris CGA009 ∆acr3 was 7.36 and 10.46 times higher than that of the R. palustris CGA009 at 100.0 µmol/L As(III) when incubated for 12 h, respectively, and 7.21 and 10.30 times higher than that of the R. palustris CGA009 when incubated with 25.0 mol/L As(V), respectively. At 25.0 mol/L As(III), low doses of methylarsonate [MAs(V)] and dimethylarsonate [DMAs(V)] were detected in both the wild type and in the deletion mutant strains. In addition, the content of As(III) in the medium changed significantly with the order being R. palustris CGA009 > R. palustris CGA009 ∆arsB > R. palustris CGA009 ∆acr3, indicating that Acr3 displayed the highest As(III) efflux rate. Conclusion: The results of this study showed that As efflux transporters were shown to be a remarkable intrinsic factor limiting As volatilization efficiency, and As volatilization rate could be significantly improved by deleting genes encoding microbial As efflux transporters. Our study provided an explanation for the often low rate of microbial As methylation and an effective strategy for screening microorganisms with high As volatilization.

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Section: Discussionmentioning

confidence: 96%

Section: Discussionmentioning

confidence: 99%

Section: However Little Information Is Available About the Genetic Dmentioning

confidence: 99%

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Arsenite efflux limits microbial arsenic volatilization

Ke¹,

Kuang²,

Zhao³

et al. 2019

Preprint

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“…Additionally, ELRs can be used as purification tags for target proteins or peptides, further simplifying the purification process ( Hassouneh et al, 2010 ). For instance, ELR has been fused to human epidermal growth factor (hEGF) ( Sarvestani et al, 2021 ), antimicrobial peptides ( Acosta et al, 2020c ; Pereira et al, 2021 ), bioactive human interferon-gamma (IFN- γ) ( Heidari-Japelaghi et al, 2020 ), and As(III) S-adenosyl-methyltransferase (ArsM) ( Ke et al, 2019 ), among others, illustrating the potential to incorporate growth factors, antimicrobial peptides, interleukins, and enzymes into our in vitro models.…”

Section: Bioproduction and Purification Elrsmentioning

confidence: 99%

Contribution of the ELRs to the development of advanced in vitro models

Puertas-Bartolomé,

Venegas-Bustos,

Acosta

et al. 2024

Front. Bioeng. Biotechnol.

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Developing in vitro models that accurately mimic the microenvironment of biological structures or processes holds substantial promise for gaining insights into specific biological functions. In the field of tissue engineering and regenerative medicine, in vitro models able to capture the precise structural, topographical, and functional complexity of living tissues, prove to be valuable tools for comprehending disease mechanisms, assessing drug responses, and serving as alternatives or complements to animal testing. The choice of the right biomaterial and fabrication technique for the development of these in vitro models plays an important role in their functionality. In this sense, elastin-like recombinamers (ELRs) have emerged as an important tool for the fabrication of in vitro models overcoming the challenges encountered in natural and synthetic materials due to their intrinsic properties, such as phase transition behavior, tunable biological properties, viscoelasticity, and easy processability. In this review article, we will delve into the use of ELRs for molecular models of intrinsically disordered proteins (IDPs), as well as for the development of in vitro 3D models for regenerative medicine. The easy processability of the ELRs and their rational design has allowed their use for the development of spheroids and organoids, or bioinks for 3D bioprinting. Thus, incorporating ELRs into the toolkit of biomaterials used for the fabrication of in vitro models, represents a transformative step forward in improving the accuracy, efficiency, and functionality of these models, and opening up a wide range of possibilities in combination with advanced biofabrication techniques that remains to be explored.

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