Development of a whole-cell biocatalyst for diisobutyl phthalate degradation by functional display of a carboxylesterase on the surface of Escherichia coli.

Bacillus / enzymology Biodegradation, Environmental Carboxylesterase / genetics Cell Surface Display Techniques Dibutyl Phthalate / analogs & derivatives Escherichia coli / genetics Recombinant Fusion Proteins / genetics
Carboxylesterase Cell surface display Diisobutyl phthalate Phthalic acid esters Whole-cell biocatalyst

Journal

Microbial cell factories
ISSN: 1475-2859
Titre abrégé: Microb Cell Fact
Pays: England
ID NLM: 101139812

Informations de publication

Date de publication:
29 May 2020
Historique:
received: 10 02 2020
accepted: 25 05 2020
entrez: 31 5 2020
pubmed: 31 5 2020
medline: 29 1 2021
Statut: epublish

Résumé

Phthalic acid esters (PAEs) are widely used as plasticizers or additives during the industrial manufacturing of plastic products. PAEs have been detected in both aquatic and terrestrial environments due to their overuse. Exposure of PAEs results in human health concerns and environmental pollution. Diisobutyl phthalate is one of the main plasticizers in PAEs. Cell surface display of recombinant proteins has become a powerful tool for biotechnology applications. In this current study, a carboxylesterase was displayed on the surface of Escherichia coli cells, for use as whole-cell biocatalyst in diisobutyl phthalate biodegradation. A carboxylesterase-encoding gene (carEW) identified from Bacillus sp. K91, was fused to the N-terminal of ice nucleation protein (inpn) anchor from Pseudomonas syringae and gfp gene, and the fused protein was then cloned into pET-28a(+) vector and was expressed in Escherichia coli BL21(DE3) cells. The surface localization of INPN-CarEW/or INPN-CarEW-GFP fusion protein was confirmed by SDS-PAGE, western blot, proteinase accessibility assay, and green fluorescence measurement. The catalytic activity of the constructed E. coli surface-displayed cells was determined. The cell-surface-displayed CarEW displayed optimal temperature of 45 °C and optimal pH of 9.0, using p-NPC This work provides a promising strategy of cost-efficient biodegradation of diisobutyl phthalate for environmental bioremediation by displaying CarEW on the surface of E. coli cells. This approach might also provide a reference in treatment of other different kinds of environmental pollutants by displaying the enzyme of interest on the cell surface of a harmless microorganism.

Sections du résumé

BACKGROUND BACKGROUND
Phthalic acid esters (PAEs) are widely used as plasticizers or additives during the industrial manufacturing of plastic products. PAEs have been detected in both aquatic and terrestrial environments due to their overuse. Exposure of PAEs results in human health concerns and environmental pollution. Diisobutyl phthalate is one of the main plasticizers in PAEs. Cell surface display of recombinant proteins has become a powerful tool for biotechnology applications. In this current study, a carboxylesterase was displayed on the surface of Escherichia coli cells, for use as whole-cell biocatalyst in diisobutyl phthalate biodegradation.
RESULTS RESULTS
A carboxylesterase-encoding gene (carEW) identified from Bacillus sp. K91, was fused to the N-terminal of ice nucleation protein (inpn) anchor from Pseudomonas syringae and gfp gene, and the fused protein was then cloned into pET-28a(+) vector and was expressed in Escherichia coli BL21(DE3) cells. The surface localization of INPN-CarEW/or INPN-CarEW-GFP fusion protein was confirmed by SDS-PAGE, western blot, proteinase accessibility assay, and green fluorescence measurement. The catalytic activity of the constructed E. coli surface-displayed cells was determined. The cell-surface-displayed CarEW displayed optimal temperature of 45 °C and optimal pH of 9.0, using p-NPC
CONCLUSIONS CONCLUSIONS
This work provides a promising strategy of cost-efficient biodegradation of diisobutyl phthalate for environmental bioremediation by displaying CarEW on the surface of E. coli cells. This approach might also provide a reference in treatment of other different kinds of environmental pollutants by displaying the enzyme of interest on the cell surface of a harmless microorganism.

Identifiants

DOI: 10.1186/s12934-020-01373-6 PMID: 32471417 PMC: PMC7260753
pubmed: 32471417
doi: 10.1186/s12934-020-01373-6
pii: 10.1186/s12934-020-01373-6
pmc: PMC7260753
doi:

Substances chimiques

Recombinant Fusion Proteins 0
Dibutyl Phthalate 2286E5R2KE
Carboxylesterase EC 3.1.1.1
diisobutyl phthalate IZ67FTN290

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

114

Subventions

Organisme : National Natural Science Foundation of China
ID : 31960023
Organisme : National Natural Science Foundation of China
ID : 31600083
Organisme : Reserve Talents Project for Young and Middle-Aged Academic and Technical Leaders of Yunnan Province
ID : 2018HB059
Organisme : National Key Research and Development Program of China
ID : 2017YFB0308401

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Auteurs

Junmei Ding (J)

Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Yunnan Normal University, Kunming, 650500, Yunnan, China. djm3417@163.com.
Key Laboratory of Yunnan for Biomass Energy and Biotechnology of Environment, Kunming, 650500, Yunnan, China. djm3417@163.com.
Key Laboratory of Enzyme Engineering, Yunnan Normal University, Kunming, 650500, Yunnan, China. djm3417@163.com.
ORCID: 0000-0002-8300-436X

Yang Zhou (Y)

Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Yunnan Normal University, Kunming, 650500, Yunnan, China.
Key Laboratory of Yunnan for Biomass Energy and Biotechnology of Environment, Kunming, 650500, Yunnan, China.
Key Laboratory of Enzyme Engineering, Yunnan Normal University, Kunming, 650500, Yunnan, China.

Chaofan Wang (C)

Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Yunnan Normal University, Kunming, 650500, Yunnan, China.
Key Laboratory of Yunnan for Biomass Energy and Biotechnology of Environment, Kunming, 650500, Yunnan, China.
Key Laboratory of Enzyme Engineering, Yunnan Normal University, Kunming, 650500, Yunnan, China.

Zheng Peng (Z)

Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Yunnan Normal University, Kunming, 650500, Yunnan, China.
Key Laboratory of Yunnan for Biomass Energy and Biotechnology of Environment, Kunming, 650500, Yunnan, China.
Key Laboratory of Enzyme Engineering, Yunnan Normal University, Kunming, 650500, Yunnan, China.

Yuelin Mu (Y)

Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Yunnan Normal University, Kunming, 650500, Yunnan, China.
Key Laboratory of Yunnan for Biomass Energy and Biotechnology of Environment, Kunming, 650500, Yunnan, China.
Key Laboratory of Enzyme Engineering, Yunnan Normal University, Kunming, 650500, Yunnan, China.

Xianghua Tang (X)

Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Yunnan Normal University, Kunming, 650500, Yunnan, China.
Key Laboratory of Yunnan for Biomass Energy and Biotechnology of Environment, Kunming, 650500, Yunnan, China.
Key Laboratory of Enzyme Engineering, Yunnan Normal University, Kunming, 650500, Yunnan, China.

Zunxi Huang (Z)

Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Yunnan Normal University, Kunming, 650500, Yunnan, China. huangzunxi@163.com.
Key Laboratory of Yunnan for Biomass Energy and Biotechnology of Environment, Kunming, 650500, Yunnan, China. huangzunxi@163.com.
Key Laboratory of Enzyme Engineering, Yunnan Normal University, Kunming, 650500, Yunnan, China. huangzunxi@163.com.

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Classifications MeSH

questionsmedicales.fr › Bactéries › Bactéries à Gram positif › Bâtonnets à Gram positif › Bâtonnets sporulés à Gram positif › Bacillaceae › Bacillus › Development of a whole-cell biocatalyst for...
questionsmedicales.fr › Environnement et santé publique › Santé publique › Pollution de l'environnement › Assainissement et restauration de l'environnement › Dépollution biologique de l'environnement › Development of a whole-cell biocatalyst for...
questionsmedicales.fr › Enzymes et coenzymes › Enzymes › Hydrolases › Esterases › Carboxylic ester hydrolases › Carboxylesterase › Development of a whole-cell biocatalyst for...
questionsmedicales.fr › Techniques d'investigation › Techniques génétiques › Génie génétique › Ingénierie des protéines › Techniques d'exposition à la surface cellulaire › Development of a whole-cell biocatalyst for...
questionsmedicales.fr › Composés chimiques organiques › Acides carboxyliques › Acides carbocycliques › Acides phtaliques › Phtalate de dibutyle › Development of a whole-cell biocatalyst for...
questionsmedicales.fr › Bactéries › Proteobacteria › Gammaproteobacteria › Enterobacteriaceae › Escherichia › Escherichia coli › Development of a whole-cell biocatalyst for...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines recombinantes › Protéines de fusion recombinantes › Development of a whole-cell biocatalyst for...