Characterization of a novel thermophilic pyrethroid-hydrolyzing carboxylesterase from Sulfolobus tokodaii into a new family

Wei, Tao; Feng, Shengxue; Shen, Yulong; He, Peixin; Ma, Guanghui; Xuan, Yue; Zhang, Fei; Mao, Duobin

doi:10.1016/j.molcatb.2013.07.022

Cited by 25 publications

(18 citation statements)

References 34 publications

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“…Mutants have higher activity than WT over a range of pH values. Similar results were obtained in most other pyrethroid hydrolases [ 16 – 21 , 23 ], very different from alkaliphilic EstSt7 [ 22 ]. Good pH adaptability is indispensable when dealing with the frequently changeable conditions during bioremediation.…”

Section: Resultssupporting

confidence: 86%

“…8 , the mutant M2 was able to efficiently degrade all the pyrethroids tested within a short time, indicating that M2 possessed broad substrate specificity. This feature was similar to most pyrethroid hydrolases [ 16 – 22 ], probably because most pyrethroid pesticides share a similar ester bond in their molecular structure. Remarkably, M2 hydrolyzed pyrethroids much more efficiently than the WT and other pyrethroid hydrolases reported [ 16 – 22 ], reaching over 98% conversion in 15 min.…”

Section: Resultssupporting

confidence: 61%

“…CMO was highly stable below 10 °C, but at temperatures above 10 °C, catalytic activity decreased slowly along with increased incubation time [23]. EstSt7 had an optimum temperature of 80 °C [22]. Nevertheless, it showed worse adaptability to low and moderate temperatures, which are frequently encountered during degradation of pyrethroid residues.

Fig.…”

Section: Resultsmentioning

confidence: 99%

“…So far, quite a few pyrethroid-degrading genes have been cloned and characterized, such as pytY , estP , pytH , pye3 , pytZ , sys410 , EstSt7 gene, and CMO gene [ 16 – 23 ]. However, no one enzyme has all the properties, such as high hydrolytic activity [ 22 , 23 ], good stability [ 16 , 23 ], and high productivity [ 17 – 21 ], which are indispensable for practical biodegradation. And almost all of them are intracellular enzymes, which require complex extraction protocols and present issues in terms of easy deactivation and low production [ 24 ].…”

Section: Introductionmentioning

confidence: 99%

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Directed evolution and secretory expression of a pyrethroid-hydrolyzing esterase with enhanced catalytic activity and thermostability

et al. 2017

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BackgroundPyrethroids are potentially harmful to human health and ecosystems. It is necessary to develop some efficient strategies to degrade pyrethroid residues. Biodegradation is generally considered as a safe, efficient, and inexpensive way to eliminate environmental contaminants. To date, although several pyrethroid-hydrolyzing esterases have been cloned, there has been no report about a pyrethroid hydrolase with high hydrolytic activity, good stability, and high productivity, indispensable enzymatic properties in practical biodegradation. Almost all pyrethroid hydrolases are intracellular enzymes, which require complex extraction protocols and present issues in terms of easy inactivation and low production.ResultsIn this study, random mutagenesis was performed on one pyrethroid-hydrolyzing esterase, Sys410, to enhance its activity and thermostability. Two beneficial mutations, A171V and D256N, were obtained by random mutagenesis and gave rise to the mutant M2. The mutant displayed ~1.5-fold improvement in the kcat/Km value and 2.46-fold higher catalytic activity. The optimal temperature was 10 °C higher than that of the wild-type enzyme (55 °C). The half-life at 40–65 °C was 3.3–310 times longer. It was surprising that M2 has a half-life of 12 h at 70 °C while Sys410 was completely inactivated at 70 °C. In addition, the desired gene was extracellularly expressed in a Pichia pastoris host system. The soluble expression level reached up to 689.7 mg/L. Remarkably, the enzyme could efficiently degrade various pyrethroids at moderate temperature for 15 min, exceeding a hydrolysis rate of 98%, which is the highest value ever reported.ConclusionsThis is the first report about random mutagenesis and secretory expression of pyrethroid-hydrolyzing esterase with high-level productivity and purity in P. pastoris. Broad substrate specificity, enhanced activity and thermostability make M2 an ideal candidate for the biodegradation of pyrethroid residues.Electronic supplementary materialThe online version of this article (doi:10.1186/s12934-017-0698-5) contains supplementary material, which is available to authorized users.

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Section: Resultssupporting

confidence: 86%

Section: Resultssupporting

confidence: 61%

Fig.…”

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Directed evolution and secretory expression of a pyrethroid-hydrolyzing esterase with enhanced catalytic activity and thermostability

et al. 2017

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“…Our results have indicated that Est3385 could decompose pyrethroids in the absence other cofactors, based on the fact that the activity of Est3385 was not inhibited by chelating reagents (Fig. 6 ), which similar as the degrading enzyme EstSt7 28 .…”

Section: Discussionmentioning

confidence: 54%

Cloning and characterization of a pyrethroid pesticide decomposing esterase gene, Est3385, from Rhodopseudomonas palustris PSB-S

Luo

Zhang

Zhou

et al. 2018

Sci Rep

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Full length open reading frame of pyrethroid detoxification gene, Est3385, contains 963 nucleotides. This gene was identified and cloned based on the genome sequence of Rhodopseudomonas palustris PSB-S available at the GneBank. The predicted amino acid sequence of Est3385 shared moderate identities (30–46%) with the known homologous esterases. Phylogenetic analysis revealed that Est3385 was a member in the esterase family I. Recombinant Est3385 was heterologous expressed in E. coli, purified and characterized for its substrate specificity, kinetics and stability under various conditions. The optimal temperature and pH for Est3385 were 35 °C and 6.0, respectively. This enzyme could detoxify various pyrethroid pesticides and degrade the optimal substrate fenpropathrin with a Km and Vmax value of 0.734 ± 0.013 mmol·l−1 and 0.918 ± 0.025 U·µg−1, respectively. No cofactor was found to affect Est3385 activity but substantial reduction of enzymatic activity was observed when metal ions were applied. Taken together, a new pyrethroid degradation esterase was identified and characterized. Modification of Est3385 with protein engineering toolsets should enhance its potential for field application to reduce the pesticide residue from agroecosystems.

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Microbial elimination of pyrethroids: specific strains and involved enzymes

Fang

Zhang

et al. 2022

Appl Microbiol Biotechnol

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Characterization of a novel thermophilic pyrethroid-hydrolyzing carboxylesterase from Sulfolobus tokodaii into a new family

Cited by 25 publications

References 34 publications

Directed evolution and secretory expression of a pyrethroid-hydrolyzing esterase with enhanced catalytic activity and thermostability

Directed evolution and secretory expression of a pyrethroid-hydrolyzing esterase with enhanced catalytic activity and thermostability

Cloning and characterization of a pyrethroid pesticide decomposing esterase gene, Est3385, from Rhodopseudomonas palustris PSB-S

Microbial elimination of pyrethroids: specific strains and involved enzymes

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