[Fe]-Hydrogenase, Cofactor Biosynthesis and Engineering.

[Fe]-hydrogenase catalyzes the heterolytic cleavage of H2 and reversible hydride transfer to methenyl-tetrahydromethanopterin. The iron-guanylylpyridinol (FeGP) cofactor is the prosthetic group of this enzyme, in which mononuclear Fe(II) is ligated with a pyridinol and two CO ligands. The pyridinol ligand fixes the iron by an acyl carbon and a pyridinol nitrogen. Biosynthetic proteins for this cofactor are encoded in the hmd co-occurring (hcg) genes. The function of HcgB, HcgC, HcgD, HcgE, and HcgF was studied by using structure-to-function analysis, which is based on the crystal structure of the proteins and subsequent enzyme assays. Recently, we reported the catalytic properties of HcgA and HcgG, novel radical S-adenosyl methionine enzymes, by using an in vitro biosynthesis assay. Here, we review the properties of [Fe]-hydrogenase and the FeGP cofactor, and the biosynthesis of the FeGP cofactor. Finally, we discuss the expected engineering of [Fe]-hydrogenase and the FeGP cofactor.

Engineering a Formate Dehydrogenase for NADPH Regeneration.

Nicotinamide adenine dinucleotide (NADH) and nicotinamide adenine dinucleotide phosphate (NADPH) constitute major hydrogen donors for oxidative/reductive bio-transformations. NAD(P)H regeneration systems coupled with formate dehydrogenases (FDHs) represent a dreamful method. However, most of the native FDHs are NAD+ -dependent and suffer from insufficient reactivity compared to other enzymatic tools, such as glucose dehydrogenase. An efficient and competitive NADP+ -utilizing FDH necessitates the availability and robustness of NADPH regeneration systems. Herein, we report the engineering of a new FDH from Candida dubliniensis (CdFDH), which showed no strict NAD+ preference by a structure-guided rational/semi-rational design. A combinatorial mutant CdFDH-M4 (D197Q/Y198R/Q199N/A372S/K371T/▵Q375/K167R/H16L/K159R) exhibited 75-fold intensification of catalytic efficiency (kcat /Km ). Moreover, CdFDH-M4 has been successfully employed in diverse asymmetric oxidative/reductive processes with cofactor total turnover numbers (TTNs) ranging from 135 to 986, making it potentially useful for NADPH-required biocatalytic transformations.

Removal of pharmaceutical active compounds in wastewater by constructed wetlands: Performance and mechanisms.

The occurrence of pharmaceutical active compounds (PhACs) in aquatic environments is a cause for concern due to potential adverse effects on human and ecosystem health. Constructed wetlands (CWs) are cost-efficient and sustainable wastewater treatment systems for the removal of these PhACs. The removal processes and mechanisms comprise a complex interplay of photodegradation, biodegradation, phytoremediation, and sorption. This review synthesized the current knowledge on CWs for the removal of 20 widely detected PhACs in wastewater. In addition, the major removal mechanisms and influencing factors are discussed, enabling comprehensive and critical understanding for optimizing the removal of PhACs in CWs. Consequently, potential strategies for intensifying CWs system performance for PhACs removal are discussed. Overall, the results of this review showed that CWs performance in the elimination of some pharmaceuticals was on a par with conventional wastewater treatment plants (WWTPs) and, for others, it was above par. Furthermore, the findings indicated that system design, operational, and environmental factors played important but highly variable roles in the removal of pharmaceuticals. Nonetheless, although CWs were proven to be a more cost-efficient and sustainable technology for pharmaceuticals removal than other engineered treatment systems, there were still several research gaps to be addressed, mainly including the fate of a broad range of emerging contaminants in CWs, identification of specific functional microorganisms, transformation pathways of specific pharmaceuticals, assessment of transformation products and the ecotoxicity evaluation of CWs effluents.

Biochar enhanced organic matter transformation during pig manure composting: Roles of the cellulase activity and fungal community.

Fungal degradation of cellulose is a key step in the conversion of organic matter in composting. This study investigated the effects of adding 10% biochar (including, prepared from corn stalk and rape stalk corresponding to CSB and RSB) on organic matter transformation in composting and determined the role of cellulase and fungal communities in the conversion of organic matter. The results showed that biochar could enhance the conversion of organic matter, especially in RSB treatment. Biochar could increase cellulase activity, and RSB could enhance 33.78% and 30.70% the average activity of cellulase compared with the control and CSB treatments in the mesophilic to thermophilic phase, respectively. The results of high throughput sequencing demonstrated that Basidiomycota dominant in mesophilic phase, and Ascomycota dominant in other phases of composting. The redundancy analysis showed that Alternaria, Thermomycees, Aspergillus, Wallemia, and Melanocarpus might be the key fungi for the degradation of organic matter, and Fusarium, Penicillium, and Scopulariopsis may promote the conversion of organic matter. Network showed that the addition of RSB changed the interactions between fungal communities and organic matter transformation, and RSB treatment enriched members of Ascomycota related to organic matter transformation and cellulase activity. These results indicated that RSB improved organic matter conversion by enhancing the role of cellulase and fungal communities.

Engineering Isopropanol Dehydrogenase for Efficient Regeneration of Nicotinamide Cofactors.

Isopropanol dehydrogenase (IPADH) is one of the most attractive options for nicotinamide cofactor regeneration due to its low cost and simple downstream processing. However, poor thermostability and strict cofactor dependency hinder its practical application for bioconversions. In this study, we simultaneously improved the thermostability (433-fold) and catalytic activity (3.3-fold) of IPADH from Brucella suis via a flexible segment engineering strategy. Meanwhile, the cofactor preference of IPADH was successfully switched from NAD(H) to NADP(H) by 1.23 × 106-fold. When these variants were employed in three typical bioredox reactions to drive the synthesis of important chiral pharmaceutical building blocks, they outperformed the commonly used cofactor regeneration systems (glucose dehydrogenase [GDH], formate dehydrogenase [FDH], and lactate dehydrogenase [LDH]) with respect to efficiency of cofactor regeneration. Overall, our study provides two promising IPADH variants with complementary cofactor specificities that have great potential for wide applications. IMPORTANCE Oxidoreductases represent one group of the most important biocatalysts for synthesis of various chiral synthons. However, their practical application was hindered by the expensive nicotinamide cofactors used. Isopropanol dehydrogenase (IPADH) is one of the most attractive biocatalysts for nicotinamide cofactor regeneration. However, poor thermostability and strict cofactor dependency hinder its practical application. In this work, the thermostability and catalytic activity of an IPADH were simultaneously improved via a flexible segment engineering strategy. Meanwhile, the cofactor preference of IPADH was successfully switched from NAD(H) to NADP(H). The resultant variants show great potential for regeneration of nicotinamide cofactors, and the engineering strategy might serve as a useful approach for future engineering of other oxidoreductases.

The Function of Two Radical-SAM Enzymes, HcgA and HcgG, in the Biosynthesis of the [Fe]-Hydrogenase Cofactor.

In the biosynthesis of the iron-guanylylpyridinol (FeGP) cofactor, 6-carboxymethyl-5-methyl-4-hydroxy-2-pyridinol (1) is 3-methylated to form 2, then 4-guanylylated to form 3, and converted into the full cofactor. HcgA-G proteins catalyze the biosynthetic reactions. Herein, we report the function of two radical S-adenosyl methionine enzymes, HcgA and HcgG, as uncovered by in vitro complementation experiments and the use of purified enzymes. In vitro biosynthesis using the cell extract from the Methanococcus maripaludis ΔhcgA strain was complemented with HcgA or precursors 1, 2 or 3. The results suggested that HcgA catalyzes the biosynthetic reaction that forms 1. We demonstrated the formation of 1 by HcgA using the 3 kDa cell extract filtrate as the substrate. Biosynthesis in the ΔhcgG system was recovered by HcgG but not by 3, which indicated that HcgG catalyzes the reactions after the biosynthesis of 3. The data indicated that HcgG contributes to the formation of CO and completes biosynthesis of the FeGP cofactor.

Engineering of an oleate hydratase for efficient C10-Functionalization of oleic acid.

Oleate hydratase catalyzes the hydration of unsaturated fatty acids, giving access to C10-functionalization of oleic acid. The resultant 10-hydroxystearic acid is a key material for the synthesis of many biomass-derived value-added products. Herein, we report the engineering of an oleate hydratase from Paracoccus aminophilus (PaOH) with significantly improved catalytic efficiency (from 33 s-1 mM-1 to 119 s-1 mM-1), as well as 3.4 times increased half-life at 30 °C. The structural mechanism regarding the impact of mutations on the improved catalytic activity and thermostability was elucidated with the aid of molecular dynamics simulation. The practical feasibility of the engineered PaOH variant F233L/F122L/T15 N was demonstrated through the pilot synthesis of 10-hydroxystearic acid and 10-oxostearic acid via an optimized multi-enzymatic cascade reaction, with space-time yields of 540 g L-1 day-1 and 160 g L-1 day-1, respectively.

A High-Throughput Screening Method for the Directed Evolution of Hydroxynitrile Lyase towards Cyanohydrin Synthesis.

Chiral cyanohydrins are useful intermediates in the pharmaceutical and agricultural industries. In nature, hydroxynitrile lyases (HNLs) are a kind of elegant tool for enantioselective hydrocyanation of carbonyl compounds. However, currently available methods for demonstrating hydrocyanation are still stalled at precise, but low-throughput, GC or HPLC analyses. Herein, we report a chromogenic high-throughput screening (HTS) method that is feasible for the cyanohydrin synthesis reaction. This method was highly anti-interference and sensitive, and could be used to directly profile the substrate scope of HNLs either in cell-free extract or fermentation clear broth. This HTS method was also validated by generating new variants of PcHNL5 that presented higher catalytic efficiency and stronger acidic tolerance in variant libraries.

Prognostic significance of myocardial fibrosis and CMR characteristics in bicuspid aortic valve with moderate and severe aortic insufficiency.

OBJECTIVES: This study attempted to evaluate the characteristics and prognostic value of myocardial fibrosis (MF) in aortic insufficiency (AI) patients with bicuspid aortic valve (BAV) or tricuspid aortic valve (TAV) by cardiac magnetic resonance (CMR). METHODS: A total of 314 adults with CMR-diagnosed AI were retrospectively retrieved. Of them, 166 patients with moderate or severe AI were included and divided into two groups: BAV group (N = 46) and a TAV group (N = 120). The presence and characteristics of MF were assessed with CMR. The patients were followed for adverse clinical events. The prognostic capability of the parameters was assessed using Cox regression model. RESULTS: LV fibrosis was more common in the BAV group than in the TAV group (65.2% vs. 45.0%; p = 0.020). There was a strong association between BAV and MF even after adjusting for clinical and imaging variables (odds ratio: 3.57; p = 0.031). Kaplan-Meier analysis showed a higher rate of clinical adverse events in AI+BAV patients with MF during a median follow-up of 4.7 years. Multivariate Cox regression analysis showed that late gadolinium enhancement (LGE) was an independent predictor of clinical adverse outcome. CONCLUSION: MF is more common in AI with BAV than with TAV and is a predictor of clinical adverse events. KEY POINTS: • The presence and extent of late gadolinium enhancement of left ventricular were more common and severer in the bicuspid aortic valve group than in the tricuspid aortic valve group in aortic insufficiency patients. • Bicuspid aortic valve was an independent factor for myocardial fibrosis in aortic insufficiency patients. • Late gadolinium enhancement could be used as an independent predictor of adverse clinical events in this population.

Characterization and biogeochemical implications of dissolved organic matter in aquatic environments.

Dissolved organic matter (DOM) is viewed as one of the most chemically active organic substances on earth. It plays vital roles in the fate, bioavailability and toxicity of aquatic exogenous chemical species (e.g., heavy metals, organic pollutants, and nanomaterials). The characteristics of DOM such low concentrations, salt interference and complexity in aquatic environments and limitations of pretreatment for sample preparation and application of characterization techniques severely limit understanding of its nature and environmental roles. This review provides a characterization continuum of aquatic DOM, and demonstrate its biogeochemical implications, enabling in-depth insight into its nature and environmental roles. A synthesis of the effective DOM pretreatment strategies, comprising extraction and fractionation methods, and characterization techniques is presented. Additionally, the biogeochemical dynamics of aquatic DOM and its environmental implications are discussed. The findings indicate the collection of representative DOM samples from water as the first and critical step for characterizing its properties, dynamics, and environmental implications. However, various pretreatment procedures may alter DOM composition and structure, producing highly variable recoveries and even influencing its subsequent characterization. Therefore, complimentary use of various characterization techniques is highly recommended to obtain as much information on DOM as possible, as each characterization technique exhibits various advantages and limitations. Moreover, DOM could markedly change the physical and chemical properties of exogenous chemical species, influencing their transformation and mobility, and finally altering their potential bioavailability and toxicity. Several research gaps to be addressed include the impact of pretreatment on the composition and structure of aquatic DOM, molecular-level structural elucidation for DOM, and assessment of the effects of DOM dynamics on the fate, bioavailability and toxicity of exogenous chemical species.

Speciation, Distribution and Risk Assessment of Metals in Sediments from a Water Body Replenished by Effluent from a Wastewater Treatment Plant.

To assess the status and risk of metal pollution in landscape water body replenished by wastewater treatment plant effluent, the distribution of metals in sediments from three regions [regulation pond (RP), wetland lake (WL), and main lake (ML)] of Harbor Lake, Tianjin, China, was characterized. Higher levels of all metals (except Cr and Pb) were observed in RP sediments. As, Cr, Cu, Ni, Pb, and Zn were mainly bound to organic matter and sulfide (F3) and residual fractions (F4), while the exchangeable and carbonate fraction (F1) and Fe/Mn oxide fraction (F2) were the dominant forms for Cd. Additionally, finding showed that the overall risk of sediments in aquatic systems was affected by both metal toxicity and metal fractionation. Thus, according to a modified risk assessment code (RAC), the potential adverse effect of metals in sediments was medium (although As, Cd, Ni, and Zn had high mobility risks based on the RAC), decreasing in the order RP > WL > ML.

Direct Access to Medium-Chain α,ω-Dicarboxylic Acids by Using a Baeyer-Villiger Monooxygenase of Abnormal Regioselectivity.

Baeyer-Villiger monooxygenases (BVMOs) are versatile biocatalysts in organic synthesis that can generate esters or lactones by inserting a single oxygen atom adjacent to a carbonyl moiety. The regioselectivity of BVMOs is essential in determining the ratio of two regioisomers for converting asymmetric ketones. Herein, we report a novel BVMO from Pseudomonas aeruginosa (PaBVMO); this has been exploited for the direct synthesis of medium-chain α,ω-dicarboxylic acids through a Baeyer-Villiger oxidation-hydrolysis cascade. PaBVMO displayed the highest abnormal regioselectivity toward a variety of long-chain aliphatic keto acids (C16 -C20 ) to date, affording dicarboxylic monoesters with a ratio of up to 95 %. Upon chemical hydrolysis, α,ω-dicarboxylic acids and fatty alcohols are readily obtained without further treatment; this significantly reduces the synthetic steps of α,ω-dicarboxylic acids from renewable oils and fats.

Discovery of Two Native Baeyer-Villiger Monooxygenases for Asymmetric Synthesis of Bulky Chiral Sulfoxides.

Two Baeyer-Villiger monooxygenases (BVMOs), designated BoBVMO and AmBVMO, were discovered from Bradyrhizobium oligotrophicum and Aeromicrobium marinum, respectively. Both monooxygenases displayed novel features for catalyzing the asymmetric sulfoxidation of bulky and pharmaceutically relevant thioethers. Evolutionary relationship and sequence analysis revealed that the two BVMOs belong to the family of typical type I BVMOs and the subtype ethionamide monooxygenase. Both BVMOs are active toward medium- and long-chain aliphatic ketones as well as various thioether substrates but are ineffective toward cyclohexanone, aromatic ketones, and other typical BVMO substrates. BoBVMO and AmBVMO showed the highest activities (0.117 and 0.025 U/mg protein, respectively) toward thioanisole among the tested substrates. Furthermore, these BVMOs exhibited distinct activity and excellent stereoselectivity toward bulky and prochiral prazole thioethers, which is a unique feature of this family of BVMOs. No native enzyme has been reported for the asymmetric sulfoxidation of bulky prazole thioethers into chiral sulfoxides. The identification of BoBVMO and AmBVMO provides an important scaffold for discovering enzymes capable of asymmetrically oxidizing bulky thioether substrates by genome mining.IMPORTANCE Baeyer-Villiger monooxygenases (BVMOs) are valuable enzyme catalysts that are an alternative to the chemical Baeyer-Villiger oxidation reaction. Although BVMOs display broad substrate ranges, no native enzymes were reported to have activity toward the asymmetric oxidation of bulky prazole-like thioether substrates. Herein, we report the discovery of two type I BVMOs from Bradyrhizobium oligotrophicum (BoBVMO) and Aeromicrobium marinum (AmBVMO) which are able to catalyze the asymmetric sulfoxidation of bulky prazole thioethers (proton pump inhibitors [PPIs], a group of drugs whose main action is a pronounced and long-lasting reduction of gastric acid production). Efficient catalysis of omeprazole oxidation by BoBVMO was developed, indicating that this enzyme is a promising biocatalyst for the synthesis of bulky and pharmaceutically relevant chiral sulfoxide drugs. These results demonstrate that the newly identified enzymes are suitable templates for the discovery of more and better thioether-converting BVMOs.

Characterizing phosphorus removal from polluted urban river water by steel slags in a vertical flow constructed wetland.

Phosphorus (P) removal in constructed wetlands (CWs) is often low unless special substrates with high sorption capacities are used. However, the use of special substrates in vertical flow (VF) CWs has not been proved to enhance P sorption. Thus, two VF wetlands were designed to evaluate the potential for enhanced P removal from polluted urban river water, one with slag as substrate and the other as a control with gravel as substrate. Findings from batch experiments showed P sorption capacities of 3.15 gP/kg and 0.81 gP/kg, respectively, for steel slag and gravel. Different organic matter fractions played different roles in P sorption, the effects of which were significant only at high concentrations. Over a 220 days' operation, the VF-slag removed 76.0% of the influent total phosphorus (TP) at 0.159 g/m(2)·d and PO4-P of 70.9% at 0.063 g/m(2)·d, whereas the VF-gravel removed 65.0% at 0.136 g/m(2)·d and 48.6% at 0.040 g/m(2)·d, respectively. Therefore, the merit of using a steel slag substrate in VF wetlands can be significant for the removal of PO4-P.

Characteristics of nitrogen and phosphorus removal by a surface-flow constructed wetland for polluted river water treatment.

The characteristics of nitrogen (N) and phosphorus (P) removal were studied during the 2-year operation of a free water surface flow wetland of 900 m² with hydraulic loading of 0.1 m/d to evaluate its potential to treat water from an urban stream polluted with municipal and industrial wastewater. Attention was focused on the removal of dissolved N and P by harvesting plants (local Phragmites australis and Typha orientalis) at the end of each growing season. According to findings, the removals of N and P increased from 47.1% and 17.6%, respectively, in the 1st year to 52.3% and 32.4%, respectively, in the 2nd year. Increments of N and P removal were largely attributable to plant biomass, which increased from an average dry weight of 1.77 kg/m² in the 1st year to 3.41 kg/m² in the 2nd year. The amount of nutrients assimilated by plants in the 2nd year was almost double that of the 1st year. Increasing biomass in the 2nd year also improved redox conditions in the substrate layer, which contributed to increasing the efficiency of N removal. Compared with T. orientalis, P. australis was more competitive and adapted to conditions in the wetland better; it regenerated more vigorously and contributed more to nutrient removal.

Hybrid constructed wetlands for highly polluted river water treatment and comparison of surface- and subsurface-flow cells.

A series of large pilot constructed wetland (CW) systems were constructed near the confluence of an urban stream to a larger river in Xi'an, a northwestern megacity in China, for treating polluted stream water before it entered the receiving water body. Each CW system is a combination of surface-and subsurface-flow cells with local gravel, sand or slag as substrates and Phragmites australis and Typha orientalis as plants. During a one-year operation with an average surface loading of 0.053 m(3)/(m(2)·day), the overall COD, BOD, NH3-N, total nitrogen (TN) and total phosphorus (TP) removals were 72.7% ± 4.5%, 93.4% ± 2.1%, 54.0% ± 6.3%, 53.9% ± 6.0% and 69.4% ± 4.6%, respectively, which brought about an effective improvement of the river water quality. Surface-flow cells showed better NH3-N removal than their TN removal while subsurface-flow cells showed better TN removal than their NH3-N removal. Using local slag as the substrate, the organic and phosphorus removal could be much improved. Seasonal variation was also found in the removal of all the pollutants and autumn seemed to be the best season for pollutant removal due to the moderate water temperature and well grown plants in the CWs.

Facilitating secretory expression of apple seed ß-glucosidase in Komagataella phaffii for the efficient preparation of salidroside.

Plant-derived ß-glucosidases hold promise for glycoside biosynthesis via reverse hydrolysis because of their excellent glucose tolerance and robust stability. However, their poor heterologous expression hinders the development of large-scale production and applications. In this study, we overexpressed apple seed ß-glucosidase (ASG II) in Komagataella phaffii and enhanced its production from 289 to 4322 U L-1 through expression cassette engineering and protein engineering. Upon scaling up to a 5-L high cell-density fermentation, the resultant mutant ASG IIV80A achieved a maximum protein concentration and activity in the secreted supernatant of 2.3 g L-1 and 41.4 kU L-1, respectively. The preparative biosynthesis of salidroside by ASG IIV80A exhibited a high space-time yield of 33.1 g L-1 d-1, which is so far the highest level by plant-derived ß-glucosidase. Our work addresses the long-standing challenge of the heterologous expression of plant-derived ß-glucosidase in microorganisms and presents new avenues for the efficient production of salidroside and other natural glycosides.

Biochar reduces bioavailability of phosphorus during swine manure composting: Roles of phoD-harboring bacterial community.

The bioavailability of phosphorus is a vital index for evaluating the quality of compost products. This study examined the effects of adding wheat straw biochar (WSB) and bamboo charcoal (BC) on the transformation of various phosphorus fractions during composting, as well as analyzing the roles of the phoD-harboring bacterial community in the transformation of phosphorus fractions. Adding WSB and BC reduced the available phosphorus content in the compost products by 35.2 % and 38.5 %, respectively. Redundancy analysis showed that the alkaline phosphatase content and pH were the most important factors that affected the transformation of phosphorus fractions. The addition of biochar resulted in changes in the composition and structures of the phoD-harboring bacteria communities during composting. In addition, the key bacterial genera that secreted alkaline phosphatase and decomposed different forms of phosphorus under WSB and BC were different compared with those under control. Network and correlation analysis demonstrated that the activities of phoD-harboring bacteria could have been enhanced by biochar to accelerate the consumption of available phosphorus, and the activities of key phosphorus-solubilizing bacteria (Lysobacter, Methylobacterium, and Saccharothrix) might be inhibited when the pH increased, thereby increasing the insoluble phosphorus content.

Engineered Glycosidase for Significantly Improved Production of Naturally Rare Vina-Ginsenoside R7.

Ginsenosides are the main bioactive ingredients in plants of the genus Panax. Vina-ginsenoside R7 (VG-R7) is one of the rare high-value ginsenosides with health benefits. The only reported method for preparing VG-R7 involves inefficient and low-yield isolation from highly valuable natural resources. Notoginsenoside Fc (NG-Fc) isolated in the leaves and stems of Panax notoginseng is a suitable substrate for the preparation of VG-R7 via specific hydrolysis of the outside xylose at the C-20 position. Here, we first screened putative enzymes belonging to the glycoside hydrolase (GH) families 1, 3, and 43 and found that KfGH01 can specifically hydrolyze the ß-d-xylopyranosyl-(1 → 6)-ß-d-glucopyranoside linkage of NG-Fc to form VG-R7. The I248F/Y410R variant of KfGH01 obtained by protein engineering displayed a kcat/KM value (305.3 min-1 mM-1) for the reaction enhanced by approximately 270-fold compared with wild-type KfGH01. A change in the shape of the substrate binding pockets in the mutant allows the substrate to sit closer to the catalytic residues which may explain the enhanced catalytic efficiency of the engineered enzyme. This study identifies the first glycosidase for bioconversion of a ginsenoside with more than four sugar units, and it will inspire efforts to investigate other promising enzymes to obtain valuable natural products.

Computer-aided discovery of a novel thermophilic laccase for low-density polyethylene degradation.

Polyethylene (PE) and industrial dyes are recalcitrant pollutants calling for the development of sustainable solutions for their degradation. Laccases have been explored for removal of contaminants and pollutants, including dye decolorization and plastic degradation. Here, a novel thermophilic laccase from PE-degrading Lysinibaccillus fusiformis (LfLAC3) was identified through a computer-aided and activity-based screening. Biochemical studies of LfLAC3 indicated its high robustness and catalytic promiscuity. Dye decolorization experiments showed that LfLAC3 was able to degrade all the tested dyes with decolorization percentage from 39% to 70% without the use of a mediator. LfLAC3 was also demonstrated to degrade low-density polyethylene (LDPE) films after eight weeks of incubation with either crude cell lysate or purified enzyme. The formation of a variety of functional groups was detected using Fourier transform infrared spectroscopy (FTIR) and X-ray photoelectron spectroscopy (XPS). Damage on the surfaces of PE films was observed via scanning electron microscopy (SEM). The potential catalytic mechanism of LfLAC3 was disclosed by structure and substrate-binding modes analysis. These findings demonstrated that LfLAC3 is a promiscuous enzyme that has promising potential for dye decolorization and PE degradation.