From: Laccases: structure, function, and potential application in water bioremediation
Laccase source | Applied enzyme form | Type of culture, ingredients and enzyme form | Application | Reaction parameters | Results obtained | Main putative mechanisms involved | References |
---|---|---|---|---|---|---|---|
Pharmaceutical compounds | |||||||
Pycnoporus sanguineus CCT-4518 | C CI | The fungus was grown in PDA solid medium for 7 days at 28 °C. Laccase extract was produced in 50 mL of liquid media, at 28 °C for 72 h and supported | Laccase removal of 17-alpha-ethynilestradiol (EE2) | Free and immobilized laccase extract (100 U/L) were mixed with 10 mL of EE2 at 10 mg/L, 10 mL of acetate buffer, pH 4 or 5 or 10 mL of distilled water, all of this at 28 °C | 80% of removal of EE2 after 24 h by the free and immobilized laccase extract at pH 4 and 5. The immobilized form had three cycles of reusability with high transformations | The laccase is able form dimers of the EE2 by polymerization of it | [49] |
Pycnoporus sanguineus | C | The Theobroma grandiflorum AW was used as Pycnoporus sanguineus laccase (Lac) inducer, cultivated for 7 days at 28 ± 2 °C and 150 rpm | Degradation of estrogens tested | 100 U/L of laccases, with 17-α-ethinylestradiol at 10 μg/mL, and 1% of inducer by 24 h | Removal 96% of estrogens after 8 h of reaction | They suggest the degradation product, with hydroxylation of estrogens | [56] |
Trametes versicolor | F | Commercial laccase powder from T. versicolor (activity ≥ 0.5 U/mg) from Sigma-Aldrich | Degradation of PhAC: diclofenac, trimethoprim, carbamazepine, and sulfamethoxazole | Selected PhAC concentrations were added to the enzyme solution in individual beakers. The beakers were incubated on a rotary shaker for 48 h at 80 rpm and 25 °C | The results of this study revealed that laccase can effectively degrade diclofenac (100%), trimethoprim (95%), carbamazepine (85%), and sulfamethoxazole (56%) | Not reported | [50] |
Trametes hirsuta | C | It was grown on PDA medium for 5 days at 28 °C and then on petri plates, pH 5 in static condition for 10 days, on Kirk’s medium. The supernatant was used | Degradation of chloramphenicol (CAP) | Different mediators like syringaldehyde, naphthol, vanillin and ABTS were added at 0.25, 0.50, 1, 3, 5 and 10 mM, to the reaction with 100 U of laccase enzyme and 10 mg/L of CAP in 0.1 M acetate buffer pH 5, by 48 h | The laccase enzyme degraded 0.5 mg/L CAP within 7 days without mediators and was efficiently degraded in the presence of laccase mediator system (syringaldehyde, vanillin, ABTS and α-naphthol) | Dehalogenation and oxidation of CAP by laccase to form chloramphenicol aldehyde which was non-toxic to the microorganisms studied | [53] |
Trametes versicolor | C | SF (500 mL) with 20 g of dried apple pomace, Tween 80 (0.1%) and moisture of 75% (w/w), inoculated with mycelia by 14 days, 30 °C with 200 mL. Enzyme from supernatant extract | Chlortetracycline (CTC) degradation | CTC at 2 mg/L, laccase dose at 0.5 IU, pH 4.5 or 6.0, and ultrasonication | 60% of CTC, considered as a recalcitrant pollutant, was removed in 2 h by ultrasonication and assisted laccase at pH 6.0. While at pH 4.5, 80% of CTC was degraded, resulting non estrogenic by products | Oxidation of C–C and C–O bonds | [51] |
Pleurotus ostreatus | FP | PDA medium at 25 °C, and added ciprofloxacin (CIP: at 100, 200, 300, 400 and 500 ppm). The enzyme was secreted | Degradation of ciprofloxacin (CIP) | Fungi growth by 14 days with 100, 200, 300, 400 and 500 ppm of CIP | Antibiotic degradation of about 68.8, 94.25 and 91.34% was estimated after 14 days of incubation at 500 ppm CIP | Not reported | [52] |
Pycnoporus sanguineus CS43f | F | STR of 10 L with 36.8% tomato juice medium, by 15 days, induced with CuSO4 and soybean oil at 48 h. LacI and LacII were purified | Degradation of endocrine disrupting chemicals (EDCs): nonylphenol and triclosan (a biocide) | EDC at 10 ppm final concentration were prepared in pH 5 McIlvaine buffer with 100 U/L laccase. Samples were tested every 30 min for 8 h at 25 °C | More than 95% removal after 8 h of treatment with 100 U/L at pH 5 | Enzyme-driven oxidation | [41] |
Plastics, personal care and herbicide compounds | |||||||
Pycnoporus sanguineus (CS43) | CI | 11-days cultures in 10-L STR in complex liquid medium at 28 °C. Crude extract enzyme immobilized | Degradation of emerging endocrine disruptor (bisphenol A) | 800 μL McIlvaine buffer (pH 3), 100 µL of ABTS (5 mM, 1.0% w/v) and 100 µL of laccase extract of P. sanguineus (CS43) | 100% degradation of bisphenol A (20 mg/L) was achieved in less than 24 h | Probably degradation ends in the formation of 4-isopropenylphenol | [42] |
Trametes versicolor BAFC 2234 | MI | 7-days cultures in 30-L STR with complex liquid medium (50% tomato juice). Purified enzymes | In vitro oxidation of phenol | The reaction mixture in 1.5-mL contained dissolved phenol (0.5 mM), 50 mM sodium citrate pH 4.5 and 0.1 U/mL laccase | 84% phenol removal in 4 h. Dark colored products partly precipitated were found | Oxidative coupling of phenoxy radicals as major pathway of phenol conversion | [43] |
Recombinant laccase from Trametes sanguineus in Trichoderma atroviride | F | Cultures grown in 50 mL, incubated for 4 days at 28 °C/150 rpm. Purified laccase | Degradation of xenobiotic compounds (phenanthrene and benzo[α]pyrene) | Phenanthrene and benzo[α]pyrene were added into supernatants up to at 10 ppm, incubated at 28 °C and shaken at 150 rpm for 24 h | 57.5 U/L of laccase in supernatant removed phenanthrene and benzo[α]pyrene (97 and 99% respectively) present in wastewater from a biofuel industry plant | Not reported | [67] |
Nicotiana tabacum expressing a laccase from Pleurotus ostreatus | C | Plants were grown for 16 days in a growing chamber at 24 °C under a photoperiod of 16:8 h (light:darkness). Enzyme secreted into rhizosphere | Phytoremediation of phenol content from olive mill wastewaters | Laccase activity of transgenic root exudates was evaluated by oxidation of 2 mM ABTS at 420 nm in 0.1 M citrate buffer pH 3.0 at 25 °C | Transgenic tobacco plants cultivated in a hydroponic solution with olive mill wastewaters were able to reduce the total phenol content up to 70% | Not reported | [92] |
Anthracophyllum discolor | MI | It was grown in Kirk liquid medium with Tween 80 or soil supplemented with Tween 80 and wheat grains. Whole cultures | Degradation of polycyclic aromatic hydrocarbons (PAH) | Cultures and 50 mg/L of PAH at 30 °C by 28 days. 10 g soil and 0.5 g wheat grains in 30 mL tubes contaminated with a 50 mg/kg of PAHs at 30 °C by 60 days | 54 up to 75% removal of phenanthrene, anthracene. fluoranthene, pyrene and benzo (a)pyrene in soil with A. discolor | Products of degradations were anthraquinone, phthalic acid, 4-hydroxy-9-fluorenone, 9-fluorenone and 4,5-dihydropyrene | [39] |
Trametes pubescens CBS 696.94 | C | 1L SF with synthetic liquid medium supplemented with dry coffee husk. 23 days static incubation at 30 °C. Crude extracts filtered | Biodegradation of a mixture of 2-chlorophenol (CP), 2,4-dichlorophenol (DCP), 2,4,6-trichlorophenol (TCP), pentachlorophenol (PCP) | Degradation of CPs during 8 h at 40 °C, 200 rpm in flasks containing 100 mL of a CP mixture, with 15 mg/L of each CP in 50 mM phosphate buffer, pH 6.0. Enzymatic extract (5 mL) and 10 U/L | Biodegradation of 100%, 99%, 82.1% and 41.1% of CP, DCP, TCP and PCP, respectively, after 4 h. The reduction in chlorophenols, allowed 90% reduction toxicity | Not reported | [44] |
Neosartorya fischeri | C | 50 mL SF with modified Czapek medium and 20 mg of asphaltenes as carbon source, at 37 °C 100 rpm, 4 weeks. Whole cultures | Metabolization and mineralization of asphaltenes (recalcitrant petroleum fraction) | Asphaltene mineralization was quantified by measuring CO2 production. Cell-free extracellular medium was solvent extracted and analyzed by GC–MS | After 11 weeks of growth, the fungus metabolize 15.5% of the asphaltenic carbon, including 13.2% transformed to CO2 | Generation of oxidized metabolites such as hydroxypyrenedione and hydroxyphenylacetic acid | [40] |
Coriolopsis rigida LPSC 232 | C | 15-days liquid cultures in modified Czapek Dox medium (0.5% peptone and 0.15 mM Cu2+) | Detoxification of water soluble fraction from ‘‘alpeorujo” (WSFA) | Reaction mixtures containing WSFA 20% (v/v) and 20 U laccase were incubated 24 h at 28 °C and 150 rpm | Reduction of free phenols from the WSFA | Oxidation of free phenols, resulting in radical formation, leading to polymerization as well as detoxification | [208] |
Trametes villosa | C F | Extracted and purified enzyme (Novozymes) | Bisphenol A (BPA) degradation | 2.2 mM BPA incubated for 1 h with 1.0 unit/mL of laccase. The reaction mixture: 0.5 mM ABTS, 0.1 M sodium acetate, pH 5.0, and an enzyme in a total volume of 1.0 ml was incubated at 37 °C | BPA was degraded by a laccase, which was extracted and purified from DeniLite, a Novozymes’ product. Transforming and important endocrine-disturbing compound | BPA was metabolized to two compounds: one with high molecular weight due to oxidative condensation, and another identified as 4-isopro-penylphenol | [54] |
Dye-based pollutants | |||||||
Pleurotus ostreatus URM 4809 | C | 250-mL SF with 50 mL of effluent with 0.05 g/L Remazol Brilliant Blue R and 107 spores/mL and incubated | Decolorization dyes used in the textile industry | Cell in a microbial fuel cell with continuous laccase synthesis; and 0.05 g/L of anthraquinone remazol brilliant blue R dye | Laccase promoted decolorization by 86% of the anthraquinone dye remazol brilliant blue R (used in the textile industry) | Not mentioned, but phytotoxicity results showed that the process did not generate detectable toxic products | [46] |
Ganoderma lucidum E47 strain | C | Solid-state fermentation in MYSA medium, pH 5.5, kept in darkness for 7 days at 25 °C. Supernatants was used as enzyme preparation. 5 compounds were tested in 0.5 L minireactor simulating an effluent | Decolorizing xanthene, azo and triarylmethane dyes | 0.1 mM of organic dyes: Bengal rose; blue black naphthol; congo red; methyl orange; bromocresol green; bromocresol purple; bromophenol blue; and phenol red, 550 nm; 100 mM potassium acetate buffer pH 4.8, 5% butyl acetate, 25 °C | The best activity-stability reached in pH 4.8 at 37 °C, decolorizing xanthene, azo and triarylmethane dyes, with selectivity on bromocresol green and bromocresol purple. Activity on effluent biotreatment | Not reported | [47] |
Oudemansiella canarii | C | SF, mycelial from petri dishes were incubated without agitation under air at 28 °C and in the absence of light by 14 days. Extract was dialyzed and partially purified | Decolorization of congo red | 50 mM acetate buffer (pH 5.5) in 250-mL 140 SF with 50 mL and containing 50 mg/L of Congo red and native 141 laccase (5 U). The mixtures were incubated at 30 °C in the dark in a rotary shaker at 100 rpm | 5 U were able to decolorize 80% of 50 mg/L Congo red within 24 h at 30 °C and pH 5.5 | Laccase acts not only on the dye chromophore group, but also that it cleaves different covalent bonds, causing an effective fragmentation of the molecule | [48] |
P. pastoris or A. thaliana expressing Lcc9 from Laccaria bicolor | F | SF in BMGY medium at 28 °C, the cells were suspended in of BMMY. Methanol was added to 1% every 24 h | Decolorization of triphenylmethane dyes, employed in industrial dyeing processes | The reaction mixture for the decolorization assay contained 0.1 mM of crystal violet, McIlvaine buffer and 50 μL of the enzyme in a total of 200 μL. ABTS, as the mediator, was added if necessary. Incubated in dark for 24 h | In the presence of ABTS, the decolorization rates of Crystal violet by laccases in P. pastoris or A. thaliana reached 90.7% and 83.6%, respectively | Not reported | [68] |
Recombinant laccase (Lcc IIIb) from Trametes versicolor expressed in Yarrowia lipolytica | C | Cultures grown in optimized PPB medium pH 7.0 at 2 L STR | Decolorization of pollutant dyes: bromocresol purple, safranin, malachite green, kristal violet, bromothymol blue, nigrosine and phenol red | Reaction mixture was composed of 10 µL of supernatant and 90 µL of a buffer prepared by dissolving 0.1 mg of each dye in 1 mL of citrate buffer at pH 3. Dye decolorization was followed spectrophotometrically | The dye decolorization rates after the first hour were 43%, 54%, 55%, 49%, 56%, 53% and 37% for bromocresol purple, safranin, malachite green, kristal violet, Bromothymol blue, nigrosine and phenol red, respectively | Not reported | [71] |
Recombinant LCC3 from Trametes trogii BAFC 463 in Pichia pastoris | C | 4-days liquid cultures induced with methanol | Synthetic dye decolorization | 50 µM of dye, citrate–phosphate buffer pH 4.5 at 30 °C, 1–10 U/mL laccase. Mediators used ρ-coumaric acid, HBT, violuric acid (200 µM) acetosyringone (10–200 µM) | 50–100% decolorizing ability of azoic, indigoid, triarylmethane, and anthraquinonic with acetosyringone within 2 h incubation at pH 6, 70 °C | Decolorization effectiveness depended on the chemical characteristics of redox mediators and dyes, and the ratio | [70] |
Trametes trogii BAFC 463 | C F | 22-days static liquid cultures in glucose (20 g/L), asparagine (3 g/L) medium with 1 mM Cu2+ | Decolorization of synthetic dyes | 19.5 U laccase per reaction, in test tubes at 30 °C with sodium acetate buffer (10 mM, pH 4.5) in a total volume of 3 mL. The effect of different salts, heavy metals, reaction temperature, pH and redox was analyzed | Laccase decolorized 85% of indigo carmine, xylidine, malachite green, gentian violet, bromophenol blue, 65% of fast blue RR and 30% of Azure B and Methylene Blue in 24 h | Direct oxidation of certain dyes and/or by the LMS | [58] |
Trametes versicolor | FP F | The fungus was maintained on 2% malt agar slants at 25 °C Commercial purified enzyme (Fluka) | Biodegradation of triphenylmethane dyes | Reactions in SF with 100 mL dye solution (150 mg/L) buffered with 1.6 mM 2,2-dimethyl succinate, pH 4.5, at 25 °C (laccase 1225 U/L), HBT (10−3 M) | Degradation dye brilliant green1 and acid green 16. resulting benzoic acid and diethylamine and 5,7-disulfo-2-naphtoic acid respectively | Oxidation of the methyl carbon of dye structure, giving stable products | [57] |
Aspergilus expressing a laccase from Myceliophthora thermophila | I | Submerged fermentation of a recombinant Aspergillus sp. A commercial formulation, DeniLite II S, from novozymes A/S covalently immobilized | Decolorization of synthetic dyes | 20 U/mL of immobilized laccase or 0.5 g in a FBR. Several dyes at 0.02% (w/v) 30 °C 0.1 M sodium acetate buffer (pH 4.5) 90 rpm | The anthraquinonic dyes acid blue 25 and acid green 27 were decolorized. The RBBR and the diazo RB-5 were only decolorized with laccase/HBT, 31 and 60%, respectively, after 24 h | Direct oxidation of certain dyes and/or by the LMS | [45] |
Recombinant lcc1 gene from Trametes trogii in Pichia pastoris | F | SF at 30 °C and STR 2 L at 25 °C cultures in phosphate buffered minimal methanol (BMM), supplemented with yeast extract or casaminoacids | Decolorization dyes (amaranth, carmoisine, cochineal red, sunset yellow, patented blue, blue indigo and alizarin red S | 1 mL (0.05 mg/mL of dye in 0.1 M sodium phosphate buffer, pH 5.0) and 1 IU of laccase with or without 1 mM redox mediator 1-hydroxybenzotriazole or violuric acid at 25 °C, plus dyes | All the dyes were decolorized up to 60% percent after 2 h with containing 1 U of Lcc1 and the redox mediator violuric acid 1 mM | Generation of a phenoxy radical resulting in the cleavage of azo linkages with nitrogen release | [62] |