Simple Whole-Cell Biodetection and Bioremediation of Heavy Metals Based on an Engineered Lead-Specific Operon

Wei, Wei; Liu, Xiangzhi; Sun, Peiqing; Wang, Xin; Zhu, Hong; Hong, Mei; Mao, Zong‐Wan; Zhao, Jing

doi:10.1021/es4046567

Cited by 137 publications

(70 citation statements)

References 38 publications

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“…Surface-anchoring motifs, including OmpA, ice nucleation protein (INP) (Li et al 2004), and auto-transporters (van Ulsen et al 2018), act as carriers to deliver the target proteins across the bacterial cell membrane. Among them, Lpp-OmpA was frequently used (Fasehee et al 2018;Wang et al 2019;Wei et al 2014) and was considered an ideal carrier protein (Lang 2000). For example, E. coli cells expressing Lpp-OmpAanchored EC20 (a metal-binding protein) accumulated substantially more mercury than did cells engineered with INP-fused EC20 (Bae et al 2002).…”

Section: Discussionmentioning

confidence: 99%

A simple and rapid protein purification method based on cell-surface display of SUMO-fused recombinant protein and Ulp1 protease

et al. 2020

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The development of novel methods for highly efficient protein purification remains a research focus in the biotechnology field because conventional purification approaches, including affinity purification, gel filtration, and ionexchange chromatography, require complex manipulation steps and are costly. Here, we describe a simple and rapid protein purification strategy in which the SUMO tag and Ulp1 protease are surface-displayed separately on Escherichia coli cells. After protein induction, the cells are harvested, resuspended in cleavage buffer, and incubated together for cleavage. In this approach, the surface-displayed Ulp1 cleaves the membrane-anchored SUMO fusion protein, resulting in the release of the target protein from the C-terminal of SUMO into the solution. The bacterial cells harboring SUMO and Ulp1 on their surfaces can be easily removed by centrifugation. To evaluate the purification method, we used red fluorescent protein (mCherry). Purified mCherry protein (7.72 ± 1.05 mg from 1 L of bacterial culture) was obtained after only 30 min of incubation. The protein purity was higher than 80%, and could be further improved (> 90%) by simple ultrafiltration. This study offers a promising and simple strategy for the purification of recombinant protein in its native form that requires only cleavage and centrifugation steps.

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Section: Discussionmentioning

confidence: 99%

A simple and rapid protein purification method based on cell-surface display of SUMO-fused recombinant protein and Ulp1 protease

et al. 2020

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“…39 This issue was addressed by fusing the signal and the first nine amino acids of a lipoprotein (Lpp) to the N-terminal region of OmpA and has been used to display many functionally diverse heterologous enzymes. 40,41 …”

Section: Molecular and Cellular Principles Of Cell Surface Displaymentioning

confidence: 99%

Engineering Novel and Improved Biocatalysts by Cell Surface Display

Smith

Khera

Wen

2015

Ind. Eng. Chem. Res.

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Biocatalysts, especially enzymes, have the ability to catalyze reactions with high product selectivity, utilize a broad range of substrates, and maintain activity at low temperature and pressure. Therefore, they represent a renewable, environmentally friendly alternative to conventional catalysts. Most current industrial-scale chemical production processes using biocatalysts employ soluble enzymes or whole cells expressing intracellular enzymes. Cell surface display systems differ by presenting heterologous enzymes extracellularly, overcoming some of the limitations associated with enzyme purification and substrate transport. Additionally, coupled with directed evolution, cell surface display is a powerful platform for engineering enzymes with enhanced properties. In this review, we will introduce the molecular and cellular principles of cell surface display and discuss how it has been applied to engineer enzymes with improved properties as well as to develop surface-engineered microbes as whole-cell biocatalysts.

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“…It is also used in firearms, fertilizers and pesticides, fuel additive and as refrigerating element after fusion (SILVA, 2014). Lead may thus occur in all environments due to its wide applications in several products (WEI et al, 2014).…”

Section: Lead: Characteristics Sources and Concentration Levels In Tmentioning

confidence: 99%

Lead in Plants: A Brief Review of Its Effects, Mechanisms Toxicological and Remediation

Silva¹,

Santos²,

Guilherme³

2015

Agrarian Academy

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Environmental pollution by toxic heavy metals is one of the most serious issues that world populations have to cope with. In fact, it occurs in industrial and in agricultural regions. Lead is one of the most toxic for humans and plants with great damage for the environment. Phytotoxic activities have been registered in scientific literature by several researchers who reported its effects in many vegetal species. In fact, lead accumulation in the soil causes damage to the development of plants due to a greater concentration in the roots and distributes itself to several tissues. When absorbed by the plant can be distributed to different tissues by means of a complex network of homeostasis, causing molecular and biochemical disorders. Despite the high toxicity of lead, some plant species have been tolerant of its presence and may be used in the decontamination of soil by phytoremediation. Current research is a revision of the literature on the ecotoxicological effects of heavy metals, especially lead, and present the current technical soil remediation contaminated by the metal. KEYWORDS: contamination, heavy metal, pollutant. CHUMBO NAS PLANTAS: UMA BREVE REVISÃO SOBRE SEUS EFEITOS,MECANISMOS TOXICOLÓGICOS E REMEDIAÇÃO RESUMO A poluição ambiental por metais pesados tóxicos é um dos graves problemas enfrentados pela população mundial, estando presente nas indústrias e nas regiões agrícolas. Dentre os metais pesados, o chumbo tem sido um dos mais tóxicos para o homem e para as plantas, causando grandes preocupações ambientais. A sua ação fitotóxica tem sido reportada na literatura científica por diversos pesquisadores, que têm verificado os seus efeitos em diversas espécies de plantas. O chumbo tende a acumular no solo provocando danos ao desenvolvimento das plantas, principalmente nos tecidos radiculares. Entretanto, uma vez absorvido pelo vegetal, pode se distribuir para os diferentes tecidos por meio de uma complexa rede de homeostase, causando distúrbios bioquímico e molecular. Apesar da alta toxicidade do chumbo, algumas espécies de plantas têm apresentado tolerância a se desenvolver em sua AGRARIAN ACADEMY, Centro Científico Conhecer -Goiânia, v.2, n.03; p. 2015 2 presença, sendo estudadas como prováveis remediadoras do solo contaminado, por meio da técnica de fitorremediação. O objetivo dessa pesquisa foi fazer uma revisão de literatura sobre os mecanismos e efeitos toxicológicos dos metais pesados, particularmente do chumbo, nas plantas, assim como apresentar técnicas atuais que buscam remediar o solo contaminado por esse metal. PALAVRAS-CHAVE: contaminação, metal pesado, poluente.

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Simple Whole-Cell Biodetection and Bioremediation of Heavy Metals Based on an Engineered Lead-Specific Operon

Cited by 137 publications

References 38 publications

A simple and rapid protein purification method based on cell-surface display of SUMO-fused recombinant protein and Ulp1 protease

A simple and rapid protein purification method based on cell-surface display of SUMO-fused recombinant protein and Ulp1 protease

Engineering Novel and Improved Biocatalysts by Cell Surface Display

Lead in Plants: A Brief Review of Its Effects, Mechanisms Toxicological and Remediation

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