Biodegradation of λ-cyhalothrin through cell surface display of bacterial carboxylesterase

Ding, Junmei; Liu, Yan; Gao, Yanxiu; Zhang, Chengbo; Xu, Baojun; Yang, Yunjuan; Wu, Qian; Huang, Zunxi

doi:10.1016/j.chemosphere.2021.133130

Cited by 13 publications

(9 citation statements)

References 49 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…4 a, left panel). The localization of CarCby-INPN also agrees with other work reported previously using the same surface-displayed strategy [ 31 , 32 ]. All the results suggested that CarCby was displayed and expressed successfully on the E. coli cell surface.…”

Section: Resultssupporting

confidence: 91%

“…3 a, lane 3). INPN was used as an anchor to construct CarCby whole cell biocatalyst [ 31 , 32 ]. To confirm that CarCby-INPN could indeed be located on the outside of the outer membrane, the recombinant plasmid pET-28a(+) / CarCby / INPN/GFP that used GFP as a reporter was constructed.…”

Section: Resultsmentioning

confidence: 99%

“…Localization of target proteins on the cell surface could be used as a whole-cell biocatalyst directly because the substrates do not need to be transported across the membrane into the cells [ 6 ]. Surface-displayed systems developed using E. coli as a host and the N-terminal of ice nucleation protein (INPN) from Pseudomonas syringae as an anchor have been successfully applied in environmental bioremediation, such as biodegradation of phthalic acid esters [ 31 ], pesticides [ 32 ], antibiotics [ 33 , 34 ], and absorption of mercury ions/lead [ 35 , 36 ]. To date, no report using a surface-displayed strategy for carbamate biodegradation has been published.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Display of a novel carboxylesterase CarCby on Escherichia coli cell surface for carbaryl pesticide bioremediation

et al. 2022

Self Cite

View full text Add to dashboard Cite

Background Carbamate pesticides have been widely used in agricultural and forestry pest control. The large-scale use of carbamates has caused severe toxicity in various systems because of their toxic environmental residues. Carbaryl is a representative carbamate pesticide and hydrolase/carboxylesterase is the initial and critical enzyme for its degradation. Whole-cell biocatalysts have become a powerful tool for environmental bioremediation. Here, a whole cell biocatalyst was constructed by displaying a novel carboxylesterase/hydrolase on the surface of Escherichia coli cells for carbaryl bioremediation. Results The carCby gene, encoding a protein with carbaryl hydrolysis activity was cloned and characterized. Subsequently, CarCby was displayed on the outer membrane of E. coli BL21(DE3) cells using the N-terminus of ice nucleation protein as an anchor. The surface localization of CarCby was confirmed by SDS–PAGE and fluorescence microscopy. The optimal temperature and pH of the engineered E. coli cells were 30 °C and 7.5, respectively, using pNPC4 as a substrate. The whole cell biocatalyst exhibited better stability and maintained approximately 8-fold higher specific enzymatic activity than purified CarCby when incubated at 30 °C for 120 h. In addition, ~ 100% and 50% of the original activity was retained when incubated with the whole cell biocatalyst at 4 ℃ and 30 °C for 35 days, respectively. However, the purified CarCby lost almost 100% of its activity when incubated at 30 °C for 134 h or 37 °C for 96 h, respectively. Finally, approximately 30 mg/L of carbaryl was hydrolyzed by 200 U of the engineered E. coli cells in 12 h. Conclusions Here, a carbaryl hydrolase-containing surface-displayed system was first constructed, and the whole cell biocatalyst displayed better stability and maintained its catalytic activity. This surface-displayed strategy provides a new solution for the cost-efficient bioremediation of carbaryl and could also have the potential to be used to treat other carbamates in environmental bioremediation.

show abstract

Section: Resultssupporting

confidence: 91%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Display of a novel carboxylesterase CarCby on Escherichia coli cell surface for carbaryl pesticide bioremediation

et al. 2022

Self Cite

View full text Add to dashboard Cite

show abstract

“…To display an enzyme on the surface of a living cell bears several advantages. First, both the substrate and the product of the enzymatic reaction do not need to cross a membrane barrier [ 21 ], so the whole cell biocatalyst exhibits high efficient biodegradation than the purified enzyme [ 22 , 23 ]. Second, the enzyme being linked to the cell can be separated from the reaction mixture by simple centrifugation, and thus separated from the product it can be transferred to a new substrate preparation resulting in multi-rounds of enzymatic conversion [ 21 ], enhancing the catalytic efficiency and stereoselectivity [ 24 , 25 ], reducing reaction costs.…”

Section: Introductionmentioning

confidence: 99%

“…Second, the enzyme being linked to the cell can be separated from the reaction mixture by simple centrifugation, and thus separated from the product it can be transferred to a new substrate preparation resulting in multi-rounds of enzymatic conversion [ 21 ], enhancing the catalytic efficiency and stereoselectivity [ 24 , 25 ], reducing reaction costs. Finally, the anchoring in a matrix, in this case, the cell envelope stabilizes the enzyme and makes it less accessible to proteolytic degradation and material adsorption resulting in continuous higher activities [ 21 ], so the whole cell biocatalyst exhibited better stability than the purified enzyme [ 22 , 26 ]. Whole-cell biocatalysts that display enzymes on the cell surface have become a solution to specific biotechnological problems and already applied in industrialization.…”

Section: Introductionmentioning

confidence: 99%

Whole-Cell Display of Phosphotransferase in Escherichia coli for High-Efficiency Extracellular ATP Production

Zhao¹,

Yang²,

Xie³

et al. 2022

Biomolecules

View full text Add to dashboard Cite

Adenosine triphosphate (ATP), as a universal energy currency, takes a central role in many biochemical reactions with potential for the synthesis of numerous high-value products. However, the high cost of ATP limits industrial ATP-dependent enzyme-catalyzed reactions. Here, we investigated the effect of cell-surface display of phosphotransferase on ATP regeneration in recombinant Escherichia coli. By N-terminal fusion of the super-folder green fluorescent protein (sfGFP), we successfully displayed the phosphotransferase of Pseudomonas brassicacearum (PAP-Pb) on the surface of E. coli cells. The catalytic activity of sfGFP-PAP-Pb intact cells was 2.12 and 1.47 times higher than that of PAP-Pb intact cells, when the substrate was AMP and ADP, respectively. The conversion of ATP from AMP or ADP were up to 97.5% and 80.1% respectively when catalyzed by the surface-displayed enzyme at 37 °C for only 20 min. The whole-cell catalyst was very stable, and the enzyme activity of the whole cell was maintained above 40% after 40 rounds of recovery. Under this condition, 49.01 mg/mL (96.66 mM) ATP was accumulated for multi-rounds reaction. This ATP regeneration system has the characteristics of low cost, long lifetime, flexible compatibility, and great robustness.

show abstract

Microbial elimination of pyrethroids: specific strains and involved enzymes

Fang

Zhang

et al. 2022

Appl Microbiol Biotechnol

View full text Add to dashboard Cite

Biodegradation of λ-cyhalothrin through cell surface display of bacterial carboxylesterase

Cited by 13 publications

References 49 publications

Display of a novel carboxylesterase CarCby on Escherichia coli cell surface for carbaryl pesticide bioremediation

Display of a novel carboxylesterase CarCby on Escherichia coli cell surface for carbaryl pesticide bioremediation

Whole-Cell Display of Phosphotransferase in Escherichia coli for High-Efficiency Extracellular ATP Production

Microbial elimination of pyrethroids: specific strains and involved enzymes

Contact Info

Product

Resources

About