Characterization of a novel amidohydrolase with promiscuous esterase activity from a soil metagenomic library and its application in degradation of amide herbicides.

Amide herbicides have been extensively used worldwide and have received substantial attention due to their adverse environmental effects. Here, a novel amidohydrolase gene was identified from a soil metagenomic library using diethyl terephthalate (DET) as a screening substrate. The recombinant enzyme, AmiH52, was heterologously expressed in Escherichia coli and later purified and characterized, with the highest activity occurring at 40 ℃ and pH 8.0. AmiH52 was demonstrated to have both esterase and amidohydrolase activities, which exhibited highly specific activity for p-nitrophenyl butyrate (2669 U/mg) and degrading activity against several amide herbicides. In particular, it displayed the strongest activity against propanil, with a high degradation rate of 84% at 8 h. A GC-MS analysis revealed that propanil was transformed into 3,4-dichloroaniline (3,4-DCA) during this degradation. The molecular interactions and binding stability were then analyzed by molecular docking and molecular dynamics simulation, which revealed that several key amino acid residues, including Tyr164, Trp66, Ala59, Val283, Arg58, His33, His191, and His226, are involved in the specific interactions with propanil. This study provides a function-driven screening method for amide herbicide hydrolase from the metagenomic libraries and a promising propanil-degrading enzyme (AmiH52) for potential applications in environmental remediation.

Measurement of an intestinal biomarker during poisonings from select toxicants and toxins.

INTRODUCTION: Intestinal toxicity can occur following ingestion of various drugs, chemicals, and toxins. Intestinal fatty acid binding protein is a cytosolic protein specific to intestinal epithelial cells released into the systemic circulation following intestinal injury. Understanding intestinal toxicity in poisoning has the potential to explain mechanisms of toxicity and gastrointestinal symptoms. METHODS: Plasma samples were retrospectively analysed for intestinal fatty acid binding protein in 25 healthy controls and in those poisoned with Gloriosa superba (n = 18), Thevetia peruviana (n = 26), organophosphates (in various solvents) (n = 17), paracetamol (n = 14), glyphosate (n = 20), 2-methyl-4-chlorophenoxyacetic acid (n = 18) and propanil (n = 19). RESULTS: Median peak plasma intestinal fatty acid binding protein concentrations were significantly higher in patients poisoned with Gloriosa superba (2,994.1 µg/L; interquartile range 600.0-5,158.2, P < 0.001), Thevetia peruviana (1,292.5 µg/L; interquartile range 760.3 - 2,076.2; P < 0.001), glyphosate (1,803.6 µg/L; interquartile range 225.7-8,927.7; P < 0.001), 2-methyl-4-chlorophenoxyacetic acid (1,236.2 µg/L; interquartile range 192.6 - 1,709.7; P = 0.010), paracetamol (1,066.5 µg/L; interquartile range 512.9 - 1,336.9; P = 0.035), and organophosphate poisoning (729.8 µg/L; interquartile range 431.5 - 1,588.2; P = 0.046) than in healthy controls (221.6 µg/L; interquartile range 134.8 - 460.1). Median intestinal fatty acid binding protein was not statistically significantly increased compared to controls after propanil poisoning (630.0 µg/L; interquartile range 23.5 - 1,390.3; P = 0.423). CONCLUSIONS: Our pilot study describes intestinal injury assessed by elevated plasma intestinal fatty acid binding protein concentrations following the ingestion of several poisons. This serves as a foundation for further exploration into enterocyte damage in toxicology.

Amide herbicides: Analysis of their environmental fate, combined plant-microorganism soil remediation scheme, and risk prevention and control strategies for sensitive populations.

In this study, we predicted the environmental fate of amide herbicides (AHs) using the EQC (EQuilibrium Criterion) model. We found that the soil phase is the main reservoir of AHs in the environment. Second, a toxicokinetic prediction indicated that butachlor have a low human health risk, while the alachlor, acetochlor, metolachlor, napropamide, and propanil are all uncertain. To address the environmental and human-health-related threats posed by AHs, 27 new proteins/enzymes that easily absorb, degrade, and mineralize AHs were designed. Compared with the target protein/enzyme, the comprehensive evaluation value of the new proteins/enzymes increased significantly: the absorption protein increased by 20.29-113.49%; the degradation enzyme increased by 151.26-425.22%; and the mineralization enzyme increased by 23.70-52.16%. Further experiments revealed that the remediating effect of 13 new proteins/enzymes could be significantly enhanced to facilitate their applicability under real environmental conditions. The hydrophobic interactions, van der Waals forces, and polar solvation are the key factors influencing plant-microorganism remediation. Finally, the simulations revealed that appropriate consumption of kiwifruit or simultaneous consumption of ginseng, carrot, and spinach, and avoiding the simultaneous consumption of maize and carrot/spinach are the most effective means reduce the risk of exhibiting AH-linked toxicity.

Wide substrate range for a candidate bioremediation enzyme isolated from Nocardioides sp. strain SG-4 G.

Narrow substrate ranges can impact heavily on the range of applications and hence commercial viability of candidate bioremediation enzymes. Here we show that an ester hydrolase from Nocardioides strain SG-4 G has potential as a bioremediation agent against various pollutants that can be detoxified by hydrolytic cleavage of some carboxylester, carbamate, or amide linkages. Previously we showed that a radiation-killed, freeze-dried preparation (ZimA) of this strain can rapidly degrade the benzimidazole fungicide carbendazim due to the activity of a specific ester hydrolase, MheI. Here, we report that ZimA also has substantial hydrolytic activity against phthalate diesters (dimethyl, dibutyl, and dioctyl phthalate), anilide (propanil and monalide), and carbamate ester (chlorpropham) herbicides under laboratory conditions. The reaction products are substantially less toxic, or inactive as herbicides, than the parent compounds. Tests of strain SG-4 G and Escherichia coli expressing MheI found they were also able to hydrolyse dimethyl phthalate, propanil, and chlorpropham, indicating that MheI is principally responsible for the above activities.

Experimental and computational approaches to characterize a novel amidase that initiates the biodegradation of the herbicide propanil in Bosea sp. P5.

The herbicide propanil and its major metabolite 3,4-dichloroaniline (3,4-DCA) are difficult to biodegrade and pose great health and environmental risks. However, studies on the sole or synergistic mineralization of propanil by pure cultured strains are limited. A two-strain consortium (Comamonas sp. SWP-3 and Alicycliphilus sp. PH-34), obtained from a swep-mineralizing enrichment culture that can synergistically mineralize propanil, has been previously reported. Here, another propanil degradation strain, Bosea sp. P5, was successfully isolated from the same enrichment culture. A novel amidase, PsaA, responsible for initial propanil degradation, was identified from strain P5. PsaA shared low sequence identity (24.0-39.7 %) with other biochemically characterized amidases. PsaA exhibited optimal activity at 30 °C and pH 7.5 and had kcat and Km values of 5.7 s-1 and 125 µM, respectively. PsaA could convert the herbicide propanil to 3,4-DCA but exhibited no activity toward other herbicide structural analogs. This catalytic specificity was explained by using propanil and swep as substrates and then analyzed by molecular docking, molecular dynamics simulation and thermodynamic calculations, which revealed that Tyr138 is the key residue that affects the substrate spectrum of PsaA. This is the first propanil amidase with a narrow substrate spectrum identified, providing new insights into the catalytic mechanism of amidase in propanil hydrolysis.

Inclusions of Pesticides by ß-Cyclodextrin in Solution and Solid State: Chlorpropham, Monuron, and Propanil.

Persistence and degradation are important factors in determining the safe use of such synthetic products, and numerous studies have been addressed to develop pesticide remediation methods aimed at ameliorating these features. In this frame, the use of different cyclodextrins (CDs) molecules has attracted considerable attention due to their well-known non-toxic nature, limited environmental impact, and capability to reduce the environmental and health risks of pesticides. CDs appear to be a valuable tool for the elimination of pesticides from polluted areas as well as for better pesticide formulations that positively influence their hydrolysis or degradation. The present work investigates the interaction between ß-cyclodextrins and three commonly used pesticides (i.e., chlorpropham, monuron, and propanil) both in solution and in the solid state by means of UV-Vis, FT-IR, and X-ray powder diffractometry. We show that such interactions result in all three cases in the formation of inclusion complexes with a 1:1 stoichiometry and binding constants (Kb) of 369.9 M-1 for chlorpropham, 292.3 M-1 for monuron, and 298.3 M-1 for propanil. We also report the energy-minimized structures in silico for each complex. Our data expand and complement the available literature data in indicating CDs as a low-cost and very effective tool capable of modulating the properties that determine the environmental fate of pesticides.

Potential Common Mechanisms of Cytotoxicity Induced by Amide Herbicides via TRPA1 Channel Activation.

The "Multi-Threat Medical Countermeasure (MTMC)" strategy was proposed to develop a single drug with therapeutic efficacy against multiple pathologies or broad-spectrum protection against various toxins with common biochemical signals, molecular mediators, or cellular processes. This study demonstrated that cytotoxicity, expression of transient receptor potential cation channel subfamily A member 1 (TRPA1) mRNA, and intracellular calcium influx were increased in A549 cells exposed to amide herbicides (AHs), in which the order of cytotoxicity was metolachlor > acetochlor > propisochlor > alachlor > butachlor > propanil > pretilachlor, based on IC50 values of 430, 524, 564, 565, 619, 831, and 2333 µM, respectively. Inhibition/knockout of TRPA1 efficiently protected against cytotoxicity, decreased TRPA1 mRNA expression, and reduced calcium influx. The results suggested that the TRPA1 channel could be a key common target for AHs poisoning. The order of TRPA1 affinity for AHs was propanil > pretilachlor > metolachlor > (propiso/ala/aceto/butachlor), based on KD values of 16.2, 309, and 364 µM, respectively. The common molecular mechanisms of TRPA1-AHs interactions were clarified, including toxicity-effector groups (benzene ring, nitrogen/oxygen-containing functional groups, halogen) and residues involved in interactions (Lys787, Leu982). This work provides valuable information for the development of TRPA1 as a promising therapeutic target for broad-spectrum antitoxins.

Can Pesticides Dissolved in Runoff and Exposed to Maturing Rice (Oryza sativa) Plants be Transferred to Seeds?

Agriculture's global challenge to feed an estimated 7.7 billion people is further exacerbated by less available cropland for production and rapidly changing climate patterns. Pesticides are often utilized to minimize crop losses due to pest infestations; however, problems arise when these chemicals are transported off production acreage, either by storm or irrigation events, and into nearby water bodies. Innovative management practices are needed to not only reduce the volume of runoff, but also to mitigate various pollutants, such as pesticides, within the runoff. One such practice being evaluated involves using rice (Oryza sativa) as a pesticide mitigation tool. While rice plants may serve as a mechanism for phytoremediation, whether the seeds harvested from exposed plants could then be utilized as a human food source is an unanswered question. Thirty round mesocosms (55 L volume; 56 cm diameter; six replicates per treatment) were established with rice and exposed to aqueous concentrations of the pesticides clomazone, propanil, or cyfluthrin, as well as a mixture of the three pesticides. Six replicates with rice and no pesticide exposure served as controls. Initial pesticide exposure took place 8 weeks post-planting and continued once a week for 5 weeks. Rice plants, unmilled seeds, and mesocosm sediment were collected from each mesocosm 2 weeks after seed formation began and analyzed for pesticide concentrations using gas chromatography. Concentrations of pesticides in unmilled seed were below detection for individual exposures of clomazone, propanil, and cyfluthrin. When rice was exposed to the pesticide mixture, the mean ± SE unmilled seed cyfluthrin concentration was 14.8 ± 1.25 µg kg-1. These small-scale, preliminary studies offer insight into the possibility of using immature rice plants as a phytoremediation tool, while harvesting its grain after plant maturation for human consumption. Further research is needed to address this question on a larger scale and with multiple pesticide mixtures.

Analysis of serum levels of organochlorine pesticides and related factors in Parkinson's disease.

BACKGROUND: There is evidence that environmental factors contribute to the onset and progression of Parkinson's disease (PD). Pesticides are a class of environmental toxins that are linked to increased risk of developing PD. However, few studies have investigated the association between specific pesticides and PD, especially in China, which was one of the first countries to adopt the use of pesticides. METHODS: In this study, serum levels of 19 pesticides were measured in 90 patients with PD and 90 healthy spouse controls. We also analyzed the interaction between specific pesticides and PD. In addition, the association between pesticides and clinical features of PD was also investigated. Finally, we investigated the underlying mechanism of the association between pesticides and PD. RESULTS: Serum levels of organochlorine pesticides, which included α-hexachlorocyclohexane (HCH), ß-HCH, γ-HCH, δ-HCH, propanil, heptachlor, dieldrin, hexachlorobenzene, p,p'-dichlorodiphenyltrichloroethane and o,p'-dichloro-diphenyl-trichloroethane were higher in PD patients than controls. Moreover, α-HCH and propanil levels were associated with PD. Serum levels of dieldrin were associated with Hamilton Depression Scale and Montreal Cognitive Assessment scores in PD patients. In SH-SY5Y cells, α-HCH and propanil increased level of reactive oxygen species and decreased mitochondrial membrane potential. Furthermore, propanil, but not α-HCH, induced the aggregation of α-synuclein. CONCLUSIONS: This study revealed that elevated serum levels of α-HCH and propanil were associated with PD. Serum levels of dieldrin were associated with depression and cognitive function in PD patients. Moreover, propanil, but not α-HCH, induced the aggregation of α-synuclein. Further research is needed to fully elucidate the effects of pesticides on PD.

Pterocarpus mildbraedii leaf extract ebbs propanil-induced oxidative and apoptotic damage in the liver of rats.

Phytochemicals derived from plant sources are well recognized as sources of pharmacologically potent drugs in the treatment of several oxidative stress-related ailments. Dichloromethane/methanol (1:1) leaf extract of Pterocarpus mildbraedii was evaluated for its possible protection against oxidative stress and apoptosis in the liver of male Wistar rats exposed to propanil (PRP). In the experimental design, olive oil served as the vehicle, and rats were grouped into control (2 mL/kg olive oil), PRP (200 mg/kg/day), Pterocarpus mildbraedii extract (200 mg/kg/day), and Pterocarpus mildbraedii extract (200 mg/kg/day)+PRP (200 mg/kg/day), and treated daily, p.o., for seven days. Oxidative stress parameters, B-cell lymphoma 2 (Bcl-2), Bcl 2-associated X protein (Bax), p53, caspases (9/3), and terminal transferase dUTP nick end labeling (TUNEL) assays were observed in all groups. Propanil significantly elevated superoxide dismutase and lipid peroxidation levels, while concomitantly depleting GSH and p53 levels. Further, PRP enhanced the expressions of caspase-9, caspase-3, Bax, and TUNEL-positive cells in the liver of rats. However, these observed alterations were reversed following treatment with Pterocarpus mildbraedii extract. Our studies suggest that Pterocarpus mildbraedii extract protected against PRP toxicity by reducing oxidative stress and attenuating critical endpoints in the intrinsic apoptotic pathway.

Anaerobic Degradation of Propanil in Soil and Sediment Using Mixed Bacterial Culture.

The widespread use of the herbicide, propanil, causes severe environmental problems. In this study, the effects of propanil on the bacterial community in a sediment slurry were determined. Moreover, the degradation of the herbicide by pure and mixed cultures was first conducted under anaerobic conditions. The results showed that propanil caused significant changes in the bacterial community under anaerobic conditions. Four bacterial strains, i.e., Geobacter sp. Pr-1, Paracoccus denitrificans Pr-2, Pseudomonas sp. Pr-3, and Rhodococcus sp. Pr-4, isolated from the an enrichment sediment slurry were the first pure cultures that degraded propanil and 3,4-dichloroaniline (3,4-DCA) under anaerobic conditions. Some individual isolates showed the slow degradation of propanil and 3,4-DCA, but the mixture of the four strains increased the degradation rates of both compounds. The mixed culture of these isolates transformed more than 90% of propanil within 10 days in liquid media with the amendment of dextrose, glucose, or acetate. The determination of degradation pathway showed that propanil was transformed to 3,4-DCA and some other products before degrading completely. This study provides valuable information on the effects of propanil on the bacterial community and the synergistic degradation of propanil under anaerobic conditions.

Pterocarpus mildbraedii (Harms) extract resolves propanil-induced hepatic injury via repression of inflammatory stress responses in Wistar rats.

Pterocarpus mildbraedii (PME) is a green leafy vegetable from the Papilionaceae family. This study evaluated the anti-inflammatory activity of PME in Wistar rats exposed to experimental hepatotoxicity using propanil (PRP), a post-emergent herbicide. Animals were grouped as control, PRP, PME, and PME + PRP. After 7 days, the levels of stress-activated protein kinases/c-Jun N-terminal kinase (SAPK/JNK), p38 mitogen-activated protein kinase (p38 MAPK), and signal transducer and activator of transcription (STAT-3) were measured in rat liver. Furthermore, myeloperoxidase (MPO) and nitric oxide (NO) levels, as well as protein expressions of nuclear factor-κB (NF-κB p65), inducible nitric oxide synthase (iNOS), and cyclo-oxygenase-2 (COX-2) were determined. Compared with PRP-treated rats, PME significantly reduced the hepatic MPO and NO levels. PME also diminished NFκB, iNOS, and COX-2 protein expressions in PRP-treated rats. This study showed that Pterocarpus mildbraedii leaves produce active principles with relevant anti-inflammatory potential. PRACTICAL APPLICATIONS: Previous studies have shown that bioactive principles contained in medicinal plants can offer protection against chemically induced inflammation. Pterocarpus mildbraedii leaves, with rich content of polyphenols, flavonoids, and essential fatty acids, could be exploited as a therapeutic agent against pesticide-induced oxidative stress and inflammation. This current study has also shown that the potential of PME as a functional food is boosted by the presence of α-linolenic acid, an omega-3-fatty acid known to possess anti-inflammatory activity. Here, we elucidated the cellular mechanisms of the anti-inflammatory action of PME.

Degradation of butachlor and propanil by Pseudomonas sp. strain But2 and Acinetobacter baumannii strain DT.

Herbicides have been extensively used globally, resulting in severe environmental pollution. Novel butachlor-degrading Pseudomonas sp. strain But2 isolated from soil can degrade butachlor regardless of the concentration and grows without a lag phase. Specific degradation was increased at 0.01-0.1 mM, and did not change significantly at higher concentrations. During degradation, 2-chloro-N-(2,6-diethylphenyl) acetamide, 2,6-diethylaniline, and 1,3-diethylbenzene were formed, which indicated that deamination occurred. Moreover, Pseudomonas sp. strains could tolerate propanil at up to 0.8 mM. The mixed bacterial culture of Pseudomonas sp. But2 and Acinetobacter baumannii DT (a propanil-degrading bacterial strain) showed highly effective biodegradation of both butachlor and propanil in liquid media and soil. For example, under treatment with the mixed culture, the half-lives of propanil and butachlor were 1 and 5 days, respectively, whereas those for the control were 3 and 15 days. The adjuvants present in herbicides reduced degradation in liquid media, but did not influence herbicide removal from the soil. The results showed that the mixed bacteria culture is a good candidate for the removal of butachlor and propanil from contaminated soils.

Novel lipophilic analogues from 2,4-D and Propanil herbicides: Biological activity and kinetic studies.

This work describes the synthesis of new lipophilic amides and esters analogues of classical organochlorides herbicides by incorporation of long-chains from fatty acids and derivatives. The new fatty esters and amides were synthesized in 96-99% and 80-89% yields, respectively. In general, all compounds tested showed superior in vitro activity than commercial herbicides against growth L. sativa and A. cepa, in ranges 86-100% of germinative inhibition. The target compounds showed, significantly more susceptible towards acid hydrolysis than 2,4-dichlorophenoxyacetic acid (2,4-D). The kinetic and NMR studies showed that the incorporation of lipophilic chains resulted in a decrease in half-life time of new herbicides compounds (1.5 h) than 2,4-D (3 h). These findings suggest the synthesis of new lipophilic herbicides as potential alternative to traditional formulations, by incorporation of long fatty alkyl chains in the molecular structure of 2,4-D, resulting in superior in vitro herbicidal activity, best degradation behavior and more hydrophobic derivatives.

Cytogenotoxic effect of propanil using the Lens culinaris Med and Allium cepa L test.

Propanil can produce methemoglobinemia, hemolytic anemia, hepatotoxicity, metabolic disorder and nephrotoxicity. It also has a genotoxic effect, although it is not listed as a carcinogen and it continues to be applied excessively throughout the world. Consequently, in this study the cytogenotoxic effect of propanil was evaluated, using apical root cells of Allium cepa and Lens culinaris. In which, L. culinaris seeds and A. cepa bulbs were subjected to 6 treatments with propanil (2, 4, 6, 8, 10 and 12 mg L-1) and to distilled water as control treatment. Subsequently, the root growth was measured every 24 h for 3 days. Next, the mitotic index and cellular anomalies were determined. Whereby, decreased root development was observed in all treatments. Likewise, greater inhibition of mitosis was evidenced in L. culinaris compared to A. cepa. In addition, chromosomal abnormalities, such as nucleus absence, sticky chromosomes in metaphase and binucleated cells, were present in most of the treatments. Thus, the presence of micronuclei and the results of L. culinaris, indicate the high cytogenotoxicity of propanil and the feasibility of this species as bioindicator.

Biodegradation of propanil by Acinetobacter baumannii DT in a biofilm-batch reactor and effects of butachlor on the degradation process.

The herbicide, propanil, has been extensively applied in weed control, which causes serious environmental pollution. Acinetobacter baumannii DT isolated from soil has been used to determine the degradation rates of propanil and 3,4-dichloroaniline by freely suspended and biofilm cells. The results showed that the bacterial isolate could utilize both compounds as sole carbon and nitrogen sources. Edwards's model could be fitted well to the degradation kinetics of propanil, with the maximum degradation of 0.027 ± 0.003 mM h-1. The investigation of the degradation pathway showed that A. baumannii DT transformed propanil to 3,4-dichloroaniline before being completely degraded via the ortho-cleavage pathway. In addition, A. baumannii DT showed high tolerance to butachlor, a herbicide usually mixed with propanil to enhance weed control. The presence of propanil and butachlor in the liquid media increased the cell surface hydrophobicity and biofilm formation. Moreover, the biofilm reactor showed increased degradation rates of propanil and butachlor and high tolerance of bacteria to these chemicals. The obtained results showed that A. baumannii DT has a high potential in the degradation of propanil.

Mineralization of the herbicide swep by a two-strain consortium and characterization of a new amidase for hydrolyzing swep.

BACKGROUND: Swep is an excellent carbamate herbicide that kills weeds by interfering with metabolic processes and inhibiting cell division at the growth point. Due to the large amount of use, swep residues in soil and water not only cause environmental pollution but also accumulate through the food chain, ultimately pose a threat to human health. This herbicide is degraded in soil mainly by microbial activity, but no studies on the biotransformation of swep have been reported. RESULTS: In this study, a consortium consisting of two bacterial strains, Comamonas sp. SWP-3 and Alicycliphilus sp. PH-34, was enriched from a contaminated soil sample and shown to be capable of mineralizing swep. Swep was first transformed by Comamonas sp. SWP-3 to the intermediate 3,4-dichloroaniline (3,4-DCA), after which 3,4-DCA was mineralized by Alicycliphilus sp. PH-34. An amidase gene, designated as ppa, responsible for the transformation of swep into 3,4-DCA was cloned from strain SWP-3. The expressed Ppa protein efficiently hydrolyzed swep and a number of other structural analogues, such as propanil, chlorpropham and propham. Ppa shared less than 50% identity with previously reported arylamidases and displayed maximal activity at 30 °C and pH 8.6. Gly449 and Val266 were confirmed by sequential error prone PCR to be the key catalytic sites for Ppa in the conversion of swep. CONCLUSIONS: These results provide additional microbial resources for the potential remediation of swep-contaminated sites and add new insights into the catalytic mechanism of amidase in the hydrolysis of swep.

Development and application of fluorescence sensor and test strip based on molecularly imprinted quantum dots for the selective and sensitive detection of propanil in fish and seawater samples.

Molecularly imprinted quantum dots (MIP-QDs) were successfully synthesized via reversed-phase microemulsion and used as the specific recognition element and signal probe of a fluorescence sensor or test strip to achieve the highly sensitive detection of propanil. The physical-chemical characteristics and excellent selectivity of MIP-QDs were elucidated. Under optimized parameters, the MIP-QDs had good linearity at the propanil concentration range of 1.0 µg/L to 20.0 × 103 µg/L by fluorescence quenching. The developed MIP-QD-based fluorescence sensor showed good recoveries ranging from 87.2 % to 112.2 %, and the relative standard deviation was below 6.0 % for the fish and seawater samples. In addition, the limits of detection (LODs) for fish and seawater were 0.42 µg/kg and 0.38 µg/L, respectively. The fluorescence test strip developed on the basis of the MIP-QDs also displayed satisfactory recoveries of 90.1 %-111.1 %, and the LOD for propanil in the seawater sample was 0.6 µg/L. The proposed fluorescence sensor and test strip were successfully used in propanil determination in environment and aquatic products.

Spirosoma sordidisoli sp. nov., a propanil-degrading bacterium isolated from a herbicide-contaminated soil.

A novel Gram-stain negative, aerobic, rod-shaped, asporogenous, propanil-degrading bacterial strain, TY50T, was isolated from a herbicide-contaminated soil in Nanjing, China. Strain TY50T was found to grow optimally at pH 9.0, 30 °C and in the absence of NaCl. The G + C content of the total DNA was determined to be 55.5 mol%. The 16S rRNA gene sequence of strain TY50T shows high identity to that of Spirosoma lacussanchae CPCC 100624T (99.3%), Spirosoma metallicum PR1014kT (94.8%) and Spirosoma soli MIMBbqt12T (94.6%). DNA-DNA hybridization indicated that the isolate had relatively low levels of DNA-DNA relatedness with S. lacussanchae CPCC 100624T (48.3%). Average nucleotide identity and the digital DNA-DNA hybridizations for draft genomes between strain TY50T and S. lacussanchae CPCC 100624T were 93.2% and 51.0%, respectively. The major cellular fatty acids of strain TY50T were identified as C16:1ω5c (24.5%) and summed feature 3 (C16:1ω6c/C16:1ω7c, 40.7%). MK-7 was found to be the predominant respiratory quinone. The major polar lipid profile includes phosphatidylethanolamine, an unidentified lipid and an unidentified aminolipid. These chemotaxonomic data support the affiliation of strain TY50T with the members of the genus Spirosoma. Strain TY50T can be distinguished from its close phylogenetic neighbours based on its phenotypic, genotypic, and chemotaxonomic features. Therefore, strain TY50T represents a novel member of the genus Spirosoma, for which the name Spirosoma sordidisoli sp. nov. is proposed. The type strain is TY50T (= KCTC 62494T = CCTCC AB 2018041T).

Characterization of an arylamidase from a newly isolated propanil-transforming strain of Ochrobactrum sp. PP-2.

Propanil, one of the most extensively used post-emergent contact herbicides, has also been reported to have adverse effect on environmental safety. A bacterial strain of Ochrobactrum sp. PP-2, which was capable of transforming propanil, was isolated from a propanil-contaminated soil collected from a chemical factory. An arylamidase gene mah responsible for transforming propanil to 3,4-dichloroaniline (3,4-DCA) was cloned from strain PP-2 by shotgun method and subsequently confirmed by function expression. The arylamidase Mah shares low amino acid sequence identity (27-50%) with other biochemically characterized amidases and shows less than 30% identities to other reported propanil hydrolytic enzymes. Mah was most active at pH 8 and 35 °C. Mah had a remarkable activity toward propanil (Km = 6.3 ±â€¯1.2 µM), showing the highest affinity efficiency for propanil as compared with other reported propanil hydrolytic enzymes. Our study also provides a new arylamidase for the hydrolysis of propanil.