Formulation and characterization of an immobilized laccase biocatalyst and its application to eliminate organic micropollutants in wastewater

Nair, Rakesh; Demarche, Philippe; Agathos, Spiros N.

doi:10.1016/j.nbt.2012.12.004

Cited by 99 publications

(73 citation statements)

References 38 publications

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“…pubescens MUT 2400 (Mycotheca Universitatis Taurinensis, Turin, Italy) was grown in a rich nutrient medium in agitated conditions (120 rpm), as described by Nair et al (2013). After the 0.2 mm filtration, the laccase crude extract was stored at 4 C until the experiments.…”

Section: Enzymatic Treatmentmentioning

confidence: 99%

Removal of micropollutants by fungal laccases in model solution and municipal wastewater: evaluation of estrogenic activity and ecotoxicity

Spina

Cordero

Schilirò

et al. 2015

Journal of Cleaner Production

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a b s t r a c tThis study describes a multidisciplinary approach that investigates the breakdown potential of a laccase mediated system from Trametes pubescens MUT 2400 against several micropollutants including already recognized endocrine disrupting chemicals at their natural residual concentrations (from mg/L up to ng/ L).In model solution, the chemical speciation focused on a mixture of 18 analytes and adopted stir bar sorptive extraction with directed in-situ derivatization followed by gas chromatography and mass spectroscopy analysis. The method's key performance parameters were evaluated in consideration of the chemical peculiarities and complexity of real wastewaters: precision, accuracy, estimated working range extended to a wide residual concentration interval (10 ng/L to100 mg/L) indicated its fitness for purpose. Laccases were extremely active towards all the target compounds, both in term of removal yields and rate. The maximal percentage of removal was obtained for 4-t-butylphenol, 2-hydroxybiphenyl, 4-noctylphenol, salicylic acid and estrone (percentage of removal above 90%). Enzymes concentration played a central role and in most of the cases, the catalyzed reactions were very fast: the initial concentration of 9 compounds was halved within the first 3 h.The laccase-mediated treatment was then applied to a municipal wastewater collected in a real wastewater treatment plant, containing at least 9 xenobiotics as drugs, pesticides, plasticizers and personal care products. Although the harsh chemical and biological conditions of the effluent influenced enzyme stability, the reaction took place, and above 70% transformation was obtained for most analytes during the 24 h experiment. Bioassays were carried out to estimate the estrogenic activity (the E-screen test and the MELN gene-reporter luciferase assay) and the ecotoxicity (Lepidium sativum, Pseudokirchneriella subcapitata and Vibrio fischeri), demonstrating the capability of laccases to mediate an effective detoxification of the wastewater and a decrease of the estrogenic activity. treatment plant; U/L, enzymatic activity expressed as International Unit per liter; PR, percentage of removal; MEC, minimal effective concentration; t 1/2 , analyte half-life; Ti, Target Ions; RSD%, relative standard deviation percentage; LOQ, limit of quantification; QCx, quality control samples; Rel.Err %, relative error percentage; COD, chemical oxygen demand; GI%, germination index; I%, inhibition percentage of the algal growth or of the bacterial bioluminescence; EEQ, 17-b-estradiol equivalent quantity; RPE %, relative proliferative effect; TRANS%, rate of luciferase gene expression.

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Section: Enzymatic Treatmentmentioning

confidence: 99%

Removal of micropollutants by fungal laccases in model solution and municipal wastewater: evaluation of estrogenic activity and ecotoxicity

Spina

Cordero

Schilirò

et al. 2015

Journal of Cleaner Production

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“…After 7 d of incubation (30 °C, 150 r/min), pre-cultures were used to inoculate all experimental cultures. The following media were tested: control medium, bran flakes 3% (w/v) (Dantán-González et al, 2008), malt extract 2% (v/v) (Trovaslet et al, 2007), tomato juice 13% (v/v) (Michniewicz et al, 2006;Junghanns et al, 2008), and glucose-bactopeptone (Nair et al, 2013). Flasks were incubated in a shaker at 30 °C with continuous agitation (150 r/min).…”

Section: Screening Culture Conditionsmentioning

confidence: 99%

“…The assay mixture containing 0.5 mmol/L ABTS (20 µl), 0.1 mol/L citrate/0.2 mol/L phosphate buffer (160 µl) at pH 3 and diluted supernatant (20 µl) was incubated at 25 °C. Oxidation of ABTS was monitored by the increase in the absorbance at 420 nm (A 420 , ε=3.6×10 4 L/(mol·cm)) using a microplate reader (Powerwave XS2, Bio-Tek, Bad Friedrichshall, Germany) (Eggert et al, 1996;Nair et al, 2013). Enzyme activities were expressed as international units (IU); under the conditions of the experiment, one IU corresponds to 1 μmol of product formed per minute.…”

Section: Laccase Activity Assaymentioning

confidence: 99%

Enhanced production of thermostable laccases from a native strain of Pycnoporus sanguineus using central composite design

Ramírez-Cavazos

Junghanns

Nair

et al. 2014

J. Zhejiang Univ. Sci. B

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Abstract:The production of thermostable laccases from a native strain of the white-rot fungus Pycnoporus sanguineus isolated in Mexico was enhanced by testing different media and a combination of inducers including copper sulfate (CuSO 4 ). The best conditions obtained from screening experiments in shaken flasks using tomato juice, CuSO 4 , and soybean oil were integrated in an experimental design. Enhanced levels of tomato juice as the medium, CuSO 4 and soybean oil as inducers (36.8% (v/v), 3 mmol/L, and 1% (v/v), respectively) were determined for 10 L stirred tank bioreactor runs. This combination resulted in laccase titer of 143 000 IU/L (2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid), pH 3.0), which represents the highest activity so far reported for P. sanguineus in a 10-L fermentor. Other interesting media resulting from the screening included glucose-bactopeptone which increased laccase activity up to 20 000 IU/L, whereas the inducers Acid Blue 62 and Reactive Blue 19 enhanced enzyme production in this medium 10 times. Based on a partial characterization, the laccases of this strain are especially promising in terms of thermostability (half-life of 6.1 h at 60 °C) and activity titers.

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“…The oxidation of 1mM ABTS in 0.1M phosphate buffer (pH=4.5) at 420 nm considering = 36 −1 −1 was followed using a spectrophotometer [33]. One unit of laccase activity (U) is defined as the amount of laccase required to consume 1 µmol of substrate in one min at 25°C.…”

Section: Laccase Activity Assaymentioning

confidence: 99%

Preparation of laccase-graphene oxide nanosheet/alginate composite: Application for the removal of cetirizine from aqueous solution

Sharifi‐Bonab

Rad

Talat-Mehrabad

2016

Journal of Environmental Chemical Engineering

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Formulation and characterization of an immobilized laccase biocatalyst and its application to eliminate organic micropollutants in wastewater

Cited by 99 publications

References 38 publications

Removal of micropollutants by fungal laccases in model solution and municipal wastewater: evaluation of estrogenic activity and ecotoxicity

Removal of micropollutants by fungal laccases in model solution and municipal wastewater: evaluation of estrogenic activity and ecotoxicity

Enhanced production of thermostable laccases from a native strain of Pycnoporus sanguineus using central composite design

Preparation of laccase-graphene oxide nanosheet/alginate composite: Application for the removal of cetirizine from aqueous solution

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