Biodegradation of fenoxaprop-P-ethyl (FE) by Acinetobacter sp. strain DL-2 and cloning of FE hydrolase gene afeH

Dong, Weiliang; Jiang, Sheng; Shi, Kaiwen; Wang, Fei; Li, Shuhuan; Zhou, Jie; Huang, Fei; Wang, Yicheng; Zheng, Yuxiao; Hou, Ying; Huang, Yan; Cui, Zhongli

doi:10.1016/j.biortech.2015.03.039

Cited by 39 publications

(24 citation statements)

References 30 publications

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“…A novel carboxylesterase encoding gene ( feh ) was cloned from R. ruber strain JPL-2 and FeH had a broader substrate spectrum and higher catalytic efficiency toward AOPP herbicides [10]. In the same year, another novel FE hydrolase/esterase, AfeH, which also hydrolyzed various AOPP herbicides and belonged to family VII of lipolytic enzymes, was characterized [9]. In the present study, QpeH from strain J-2 had 7.9 and 9.3% similarity to that of FeH and ChbH, respectively, but only showed 5.8% similarity to AfeH.…”

Section: Resultsmentioning

confidence: 99%

“…The QPE hydrolase gene from Pseudomonas sp. strain J-2 was cloned by first constructing a genomic DNA library using the shotgun method [9]. Bacterial genomic DNA was prepared using a high-salt extraction method [15] and subjected to partial digestion with the restriction enzyme Sau 3AI.…”

Section: Methodsmentioning

confidence: 99%

“…To date, only three QPE-degrading bacterial strains have been documented, including Pseudomonas azotoformans QDZ-1 [8], Acinetobacter sp. strain DL-2 [9], and Rhodococcus ruber JPL-2 [10]. Moreover, several genes encoding AOPP herbicide hydrolases that are involved in the degradation of AOPP herbicides have been identified [8–10].…”

Section: Introductionmentioning

confidence: 99%

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Purification and properties of a novel quizalofop-p-ethyl-hydrolyzing esterase involved in quizalofop-p-ethyl degradation by Pseudomonas sp. J-2

Zhang

et al. 2017

Microb Cell Fact

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Quizalofop-p-ethyl (QPE) is a post-emergence herbicide that effectively controls grass weeds and is often detected in the environment. However, the biochemical and molecular mechanisms of QPE degradation in the environment remains unclear. In this study, a highly effective QPE-degrading bacterial strain J-2 was isolated from acclimated activated sludge and identified as a Pseudomonas sp., containing the QPE breakdown metabolite quizalofop acid (QA) identified by Liquid Chromatography-Ion Trap-Mass Spectrometry (LC-IT-MSn) analysis. A novel QPE hydrolase esterase-encoding gene qpeH was cloned from strain J-2 and functionally expressed in Escherichia coli BL21 (DE3). The specific activity of recombinant QpeH was 198.9 ± 2.7 U mg−1 for QPE with K m and K cat values of 41.3 ± 3.6 μM and 127.3 ± 4.5 s−1. The optimal pH and temperature for the recombinant QpeH were 8.0 and 30 °C, respectively and the enzyme was activated by Ca2+, Cd2+, Li+, Fe3+ and Co2+ and inhibited by Ni2+, Fe2+, Ag+, DEPC, SDS, Tween 80, Triton X, β-mercaptoethanol, PMSF, and pCMB. In addition, the catalytic efficiency of QpeH toward different AOPP herbicides in descending order was as follows: fenoxaprop-P-ethyl > quizalofop-P-tefuryl > QPE > haloxyfop-P-methyl > cyhalofopbutyl > clodinafop-propargyl. On the basis of the phylogenetic analysis and multiple sequence alignment, the identified enzyme QpeH, was clustered with esterase family V, suggesting a new member of this family because of its low similarity of amino acid sequence with esterases reported previously.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Purification and properties of a novel quizalofop-p-ethyl-hydrolyzing esterase involved in quizalofop-p-ethyl degradation by Pseudomonas sp. J-2

Zhang

et al. 2017

Microb Cell Fact

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“…For reactivation, the metal-free enzyme was incubated with divalent metal ions (Fe 2+ , Ba 2+ , Cu 2+ , Co 2+ , Ni 2+ , Zn 2+ , Ca 2+ , Mg 2+ and Mn 2+ ) at a final concentration of 1 mM for 10 min, followed by the remaining activity determination. The procedure used to assess the metal ions was also used to determine the effects of chemical agents (Triton X-100, SDS, Tween20, Tween80 and urea) and organic reagents (methanol, ethanol, acetonitrile, isopropanol and DMSO) [22, 23]. The activity in the absence of any additives was used as the control (100%).…”

Section: Methodsmentioning

confidence: 99%

Heterologous expression of cyclodextrin glycosyltransferase from Paenibacillus macerans in Escherichia coli and its application in 2-O-α-D-glucopyranosyl-L-ascorbic acid production

Jiang

Zhou

et al. 2018

BMC Biotechnol

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BackgroundCyclodextrin glucanotransferase (CGTase) can transform L-ascorbic acid (L-AA, vitamin C) to 2-O-α-D-glucopyranosyl-L-ascorbic acid (AA-2G), which shows diverse applications in food, cosmetic and pharmaceutical industries.ResultsIn this study, the cgt gene encoding α-CGTase from Paenibacillus macerans was codon-optimized (opt-cgt) and cloned into pET-28a (+) for intracellular expression in E. coli BL21 (DE3). The Opt-CGT was purified by Ni2+-NTA resin with a 55% recovery, and specific activity was increased significantly from 1.17 to 190.75 U·mg− 1. In addition, the enzyme was adopted to transform L-AA into 9.1 g/L of AA-2G. Finally, more economic substrates, including β-cyclodextrin, soluble starch, corn starch and cassava starch could also be used as glycosyl donors, and 4.9, 3.5, 1.3 and 1.5 g/L of AA-2G were obtained, respectively.ConclusionsN-terminal amino acid is critical to the activity of CGTase suggested by its truncation study. Furthermore, when the Opt-CGT was flanked by His6-tags on the C- and N-terminal, the recovery of purification by Ni2+-NTA resin is appreciably enhanced. α-cyclodextrin was the ideal glycosyl donor for AA-2G production. In addition, the selection of low cost glycosyl donors would make the process of AA-2G production more economically competitive.

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“…SDS-PAGE was used to detect the expression of the recombinant enzyme CbaA. Recombinant CbaA was purified with Ni 2ϩ -nitrilotriacetic acid (NTA) resin (Qiagen, Valencia, CA, USA) with 0 to 300 mM imidazole in buffer A (41). The ability of the enzyme to degrade CDHB was tested in a resting cell assay, as described below.…”

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confidence: 99%

Metabolic Pathway Involved in 6-Chloro-2-Benzoxazolinone Degradation by Pigmentiphaga sp. Strain DL-8 and Identification of the Novel Metal-Dependent Hydrolase CbaA

Dong

Wang

Huang

et al. 2016

Appl Environ Microbiol

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ABSTRACT6-Chloro-2-benzoxazolinone (CDHB) is a precursor of herbicide, insecticide, and fungicide synthesis and has a broad spectrum of biological activity. Pigmentiphaga sp. strain DL-8 can transform CDHB into 2-amino-5-chlorophenol (2A5CP), which it then utilizes as a carbon source for growth. The CDHB hydrolase (CbaA) was purified from strain DL-8, which can also hydrolyze 2-benzoxazolinone (BOA), 5-chloro-2-BOA, and benzamide. The specific activity of purified CbaA was 5,900 U · mg protein ؊1for CDHB, with K m and k cat values of 0.29 mM and 8,500 s ؊1 , respectively. The optimal pH for purified CbaA was 9.0, the highest activity was observed at 55°C, and the inactive metal-free enzyme could be reactivated by Mg 2؉ , Ni 2؉ , Ca 2؉ , or Zn 2؉ . Based on the results obtained for the CbaA peptide mass fingerprinting and draft genome sequence of strain DL-8, cbaA (encoding 339 amino acids) was cloned and expressed in Escherichia coli BL21(DE3). CbaA shared 18 to 21% identity with some metal-dependent hydrolases of the PF01499 family and contained the signature metal-binding motif Q 127 XXXQ 131 XD 133 XXXH 137 . The conserved amino acid residues His288 and Glu301 served as the proton donor and acceptor. E. coli BL21(DE3-pET-cbaA) resting cells could transform 0.2 mM CDHB into 2A5CP. The mutant strain DL-8⌬cbaA lost the ability to degrade CDHB but retained the ability to degrade 2A5CP, consistent with strain DL-8. These results indicated that cbaA was the key gene responsible for CDHB degradation by strain DL-8. IMPORTANCE2-Benzoxazolinone (BOA) derivatives are widely used as synthetic intermediates and are also an important group of allelochemicals acting in response to tissue damage or pathogen attack in gramineous plants. However, the degradation mechanism of BOA derivatives by microorganisms is not clear. In the present study, we reported the identification of CbaA and metabolic pathway responsible for the degradation of CDHB in Pigmentiphaga sp. DL-8. This will provide microorganism and gene resources for the bioremediation of the environmental pollution caused by BOA derivatives. 6-Chloro-2-benzoxazolinone (CDHB) is the precursor of fenoxaprop-p-ethyl (FE) and metamifop synthesis (1). FE is a postemergence aryloxyphenoxy propionate (AOPP) herbicide that is used to control annual and perennial weeds in crops, such as soybean, turf, and wheat (2, 3). Many studies have demonstrated that FE can be hydrolyzed to fenoxaprop acid (FA) and further transformed into CDHB and 2-(4-hydroxyphenoxy)-propionic acid (HPP) by soil microorganisms (4-6). Recently, the hydrolysis of FE to FA by several esterases from different microorganisms has been reported (7-9). However, the underlying genes and the metabolic pathway responsible for the degradation of CDHB in microorganisms are unknown.CDHB is a type of 2-benzoxazolinone (BOA) derivative. BOA is widely used as a synthetic intermediate of related derivatives and is an important group of allelochemicals that respond to tissue damage or pathogen attack in gramineous plants, s...

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Biodegradation of fenoxaprop-P-ethyl (FE) by Acinetobacter sp. strain DL-2 and cloning of FE hydrolase gene afeH

Cited by 39 publications

References 30 publications

Purification and properties of a novel quizalofop-p-ethyl-hydrolyzing esterase involved in quizalofop-p-ethyl degradation by Pseudomonas sp. J-2

Purification and properties of a novel quizalofop-p-ethyl-hydrolyzing esterase involved in quizalofop-p-ethyl degradation by Pseudomonas sp. J-2

Heterologous expression of cyclodextrin glycosyltransferase from Paenibacillus macerans in Escherichia coli and its application in 2-O-α-D-glucopyranosyl-L-ascorbic acid production

Metabolic Pathway Involved in 6-Chloro-2-Benzoxazolinone Degradation by Pigmentiphaga sp. Strain DL-8 and Identification of the Novel Metal-Dependent Hydrolase CbaA

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