Immobilizing laccase on kaolinite and its application in treatment of malachite green effluent with the coexistence of Cd (П)
Graphical abstract
Introduction
Immobilization could improve laccase properties and it is an effective way to overcome the application limitations of laccase, such as low stability and high production costs (Mohamad et al., 2015, Prasad and Palanivelu, 2015). The exploration of immobilization methods of laccase has been predominantly focused (Barbosa et al., 2013, Guzik et al., 2014). The immobilization methods can increase the stability of laccases, thus significantly reducing the cost burden (Datta et al., 2013, Sheldon and van Pelt, 2013).
In the laccase immobilization process, various carriers have been reported to immobilize laccase successfully (An et al., 2015, Tan et al., 2015). Among various carriers, kaolinite as an alumino-silicate mineral is cost-efficient, facility of reusability, low mass transfer resistance and microbial corrosion resistance (Abdul Rahman et al., 2005, Hu et al., 2007). Kaolinite has negative sites on the basal surface owing to isomorphic substitution and amphoteric sites on the edge surface (Liang et al., 2017, Shu et al., 2016). The amphoteric sites are conditionally charged and pH dependent because a net positive or net negative charge can be produced due to proton adsorption (An et al., 2015, Zhang et al., 2015). Kaolinite has a low permanent charge and a significant variable charge (Sinegani et al., 2005, Xu et al., 2012a, Xu et al., 2012b). Attributing to these distinctive characters, kaolinite has rather high adsorption ability. Kaolinite is widely used in adsorption studies.
Malachite green (MG) was produced from the textile staining, aquaculture, food and medical domains (Chen et al., 2015, Sinha and Osborne, 2016). The MG belongs to persistent contaminant, and it can be readily adsorbed on solid or absorbed by organism thus leading to the accumulations in organisms (Gong et al., 2009). The accumulation of MG hinders organisms' growth, reproduction and development, and they can generate mutagenic and carcinogenic influence. Furthermore, the MG effluent is always released to environment combining with heavy metals like Cd (П) in realistic situations (Deng et al., 2013, Jasinska et al., 2012). The Cd (П) has been listed as one of the top toxic heavy metals since it can cause cancer, bone lesions, lung insufficiency, anemia, hypertension and weight loss (Long et al., 2011, Wan et al., 2018). Elevated level of Cd (П) could lead to the acute and chronic disorders in nervous, kidney, liver and cardiovascular system, therefore, efficient removal of Cd (П) makes sense (Tang et al., 2014, Wu et al., 2017). The mixing of Cd (П) makes the MG effluent more difficult to treat (Ren et al., 2018, Xu et al., 2012a, Xu et al., 2012b).
The redox-mediated bio-oxidation of MG, catalyzed by immobilized laccase, is a current technology for MG degradation (Kuhar et al., 2015, Zhang et al., 2016). Utilizing laccase immobilized on numerous carriers to catalyze MG degradation is an effective and straight forward method (Zhou et al., 2018). There are varieties of carriers, including chitosan beads, PAN/O-MMT composite nanofibers, a sponge-like hydrogel, and amino-functionalized magnetic nanoparticles were applied to immobilize laccase to degrade the MG effluent (Kumar et al., 2014, Li et al., 2016, Sun et al., 2015, Zheng et al., 2016). However, immobilizing laccase from Trametes versicolor on kaolinite to degrade MG effluent with the coexistence of Cd (II) was not fully explored. The existence of Cd (II) does have effect on the activity of free laccase and the degradation of MG by laccase (Cheng et al., 2016a, Cheng et al., 2016b). Furthermore, during the laccase immobilization process, immobilization carrier also has important effect on laccase activity and stability (Zheng et al., 2016). Hence, whether if utilizing kaolinite to immobilize laccase from Trametes versicolor could degrade MG effectively, if the coexistence of Cd (II) could affect the degradation of MG and what is the remove efficiency of Cd (II) by immobilized laccase on kaolinite should be fully explored.
In the current study, kaolinite that has rather high adsorption ability was used for laccase physical adsorption immobilization. The efficiency of loaded laccase on kaolinite was characterized by relative activity (%) and stability studies. Kaolinite-laccase (Kaolin-Lac) was applied in continuous treatment of MG effluent mixed with Cd (П) in the presence of redox mediators SA to explore the removal efficiency of MG and Cd (II) by Kaolin- Lac, the effect of Cd (II) on the degradation efficiency of MG by Kaolin- Lac. The detailed characterizations changes of kaolinite, Kaolin-Lac, Kaolin-Lac after treatment were carried out by FT-IR, SEM-EDS.
Section snippets
Materials
Laccase from Trametes versicolor, 3, 5-Dimethoxy-4-hydroxybenzaldehyde(SA) and 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) were obtained from Sigma-Aldrich. Malachite Green (MG) and Kaolinite were provided by Sinopharm Chemical Reagent Shanghai (China). All other chemicals were of analytical grade and were used as received without further purification.
Enzyme activity assay
Laccase activity was tested by analyzing the product formation rate of ABTS at the absorbance of 420 nm. One unit of laccase
SEM and elements analysis
The SEM aimed to observe possible morphological changes of kaolinite, Kaolin-Lac, and Kaolin-Lac after degradation. Kaolinite, it is a 1:1 type swelling clay (Huang et al., 2017a, Shu et al., 2016). The kaolinite lattice layers are made up of tetrahedral SiO and octahedral AlO. They are connected with the van der waals' forces. The structure lamella is filled with commutative cations and water molecules (Huang et al., 2018a, Shu et al., 2014). They can be replaced by ion exchange and inter
Conclusion
This study demonstrated the gradual progress of immobilizing laccase on kaolinite. Successful preparation of Kaolin-Lac was confirmed by structural characterizations using FT-IR, SEM-EDS. The stabilities of Kaolin-Lac were enhanced compared to free laccase. The Kaolin-Lac retained 50% of the original activity and nearly 80% decolorization for MG after 5 cycles. In the presence of mediator SA, the Kaolin-Lac used to degrade MG exhibited nearly 100% in 300 min, and almost 25% removal for Cd (П).
Acknowledgements
This study was financially supported by the Program for the National Natural Science Foundation of China (51521006, 51579098, 5110916, 51378190, 51278176, 51408206), The Natural Science Foundation of Hunan province (2018JJ3549), the National Program for Support of Top-Notch Young Professionals of China (2014), the Fundamental Research Funds for the Central Universities, the Hunan Provincial Science and Technology Plan Project (2017SK2361, 2016RS3026), the Hunan Province Water Conservancy
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These authors contribute equally to this article.