Jiayang Liu

FUNGAL PRETREATMENT OF SWITCHGRASS FOR IMPROVED SACCHARIFICATION AND SIMULTANEOUS ENZYME PRODUCTION

This study investigates fungal pretreatment of switchgrass involving solid state fermentation (SSF) to improve saccharification and simultaneously produce enzymes as co-products. The results revealed that the fungus Pycnoporus sp. SYBC-L3 can significantly degrade lignin and enhance enzymatic hydrolysis efficiency. After a 36-d cultivation period, nearly 30% reduction in lignin content was obtained without significant loss of cellulose and hemicellulose, while a considerable amount of laccase, as high as 6.3 U/g, was produced. After pretreatment, pores on switchgrass surface were observed using scanning electron microscopy (SEM). The enzymatic hydrolysis efficiency for the switchgrass with 36-d pretreatment was about 50% greater than the untreated one. Our results suggest that solid state fungal cultivation may be a good method for switchgrass pretreatment, which can simultaneously achieve high efficiency of enzymatic hydrolysis and production of some useful enzymes for other industrial utilization.

Adsorption of methylene blue on an agro-waste oiltea shell with and without fungal treatment

A lignocellulosic waste oiltea shell (OTS) was evaluated as an inexpensive sorbent to remove methylene blue (MB) from aqueous solution. Fungal treatment of OTS increased the MB adsorption by modifying the physicochemical properties of OTS and simultaneously produced laccase as a beneficial co-product. Without fungal treatment, the maximum amount of adsorption (qm) of MB by OTS was 64.4 mg/g, whereas the treatment with fungus Pycnoporus sp. and Trametes versicolor increased qm up to 72.5 mg/g and 85.7 mg/g, respectively. This is because of the improved surface area and pore sizes as well as altered chemical compositions. The equilibrium sorption data for OTS both with and without treatment fitted to the Langmuir model, and the sorption rate data well fitted to the pseudo second-order kinetic model. The changes in free energy (ΔG°) and separation factor (RL) indicated that the sorption was spontaneous and favorable. Scanning electron microscopy and Fourier transform infrared spectroscopy showed the changes in the surface morphology and functional groups of OTS after fungal treatment. The agro-waste OTS could be utilized as a low-cost adsorbent for efficient dye removal, and fungal treatment can serve as a mild and clean technique to increase the adsorptive capacity of OTS.

Effect of solid state fermentation of peanut shell on its dye adsorption performance

The effect of solid state fermentation of peanut shell to produce beneficial laccase and on its dye adsorption performance was evaluated. The resulting residues from solid fermentation were tested as sorbents (designated as SFs) in comparison to the raw peanut shell (RPS) for their ability to remove crystal violet from water. The fermentation process reduced the adsorption capacity (qm) of SF by about 50%, and changed the sorptive behavior when compared to the RPS. The Langmuir model was more suitable for fitting adsorption by SFs. qm was positively correlated with the surface area of peanut shell, but negatively correlated with acid detergent lignin content. For all the sorbents tested, the process was spontaneous and endothermic, and the adsorption followed both the pseudo 1st and 2nd order kinetic model and the film diffusion model. Dye adsorption efficiency was greater when SFs dispersed solution than when placed in filter packets.

Cloning, Expression and 3D Structure Prediction of Chitinase from Chitinolyticbacter meiyuanensis SYBC-H1.

Two CHI genes from Chitinolyticbacter meiyuanensis SYBC-H1 encoding chitinases were identified and their protein 3D structures were predicted. According to the amino acid sequence alignment, CHI1 gene encoding 166 aa had a structural domain similar to the GH18 type II chitinase, and CHI2 gene encoding 383 aa had the same catalytic domain as the glycoside hydrolase family 19 chitinase. In this study, CHI2 chitinase were expressed in Escherichia coli BL21 cells, and this protein was purified by ammonium sulfate precipitation, DEAE-cellulose, and Sephadex G-100 chromatography. Optimal activity of CHI2 chitinase occurred at a temperature of 40 °C and a pH of 6.5. The presence of metal ions Fe3+, Fe2+, and Zn2+ inhibited CHI2 chitinase activity, while Na+ and K+ promoted its activity. Furthermore, the presence of EGTA, EDTA, and β-mercaptoethanol significantly increased the stability of CHI2 chitinase. The CHI2 chitinase was active with p-NP-GlcNAc, with the Km and Vm values of 23.0 µmol/L and 9.1 mM/min at a temperature of 37 °C, respectively. Additionally, the CHI2 chitinase was characterized as an N-acetyl glucosaminidase based on the hydrolysate from chitin. Overall, our results demonstrated CHI2 chitinase with remarkable biochemical properties is suitable for bioconversion of chitin waste.

Novel approach for alleviation of soil water repellency using a crude enzyme extract from fungal pretreatment of switchgrass

Soil water repellency (SWR) caused by organic coatings on soil particles can lead to serious loss in crop production and turfgrass quality. In laboratory experiments, we tested the novel concept of direct application of enzymes to alleviate SWR. In a biofuel research project on fungal pre-treatment of switchgrass (Panicum virgatum L.) for improved saccharification, enzymatic co-products (mainly laccase mixed with other trace enzymes) were produced based on fermentation periods of 18, 36, 54, and 72 days. We characterised enzyme activities of the 18–72-day crude enzyme extracts (CEE) and applied undiluted or diluted solutions (dilutions of 5-, 10-, and 100-fold) to eight air-dried, SWR soils from several golf courses. These soils exhibited water drop penetration times (WDPT) of 345–7439 s (i.e. moderately to very strongly hydrophobic) and all showed a large decrease in SWR to WDPT <60 s after application of undiluted CEE and various dilutions of CEE for 3 days with a 1 : 1 soil : solution ratio (10 g air-dried soil and 10 mL CEE solution). The observed decrease in WDPT was positively related to increased enzyme activity level for each soil in an exponential or logarithmic relationship. Most of the improvement in SWR was observed within 1 day. Enzyme activity was maintained to varying degrees in the soil solution for up to 5 days. These preliminary results suggest that it may be feasible to use direct enzyme application from CEE, as a biomass fermentation byproduct, for remediation of hydrophobic soils, which could also offer a cost benefit for biomass fermentation.

Using Natural Biomacromolecules for Adsorptive and Enzymatic Removal of Aniline Blue from Water

The present study investigated the adsorptive and enzymatic removal of aniline blue dye (AB) from aqueous solution using waxy riceprocessing waste (RW), peanut shell (PS), microbial waste of Aspergillus niger (MW) as low cost adsorbents, and laccase (Lac) as a biocatalyst. Commercial activated carbon (AC) was also employed to compare the adsorption performance with the three adsorbents. Dye removal was examined under various parameters in batch experiments. It was found that dye removal by RW and Lac was 89–94% noticeably better than that by MW and PS (20–70%). In any cases, AC produced the highest dye removal among the tested materials. The kinetics, isotherms, and thermodynamics were then analyzed to elucidate the adsorption process by the four adsorbents. The pseudo-second order kinetic was superior to the pseudo first order kinetic model in describing adsorption for all adsorbents. The Langmuir model fitted the adsorption process very well, indicating monolayer coverage of dyes on a solid surface. A thermodynamic analysis of enthalpy (ΔH°), entropy (ΔS°), and Gibbs free energy (ΔG°) classified the adsorption as a nonspontaneous and endothermic process. The results reveal diverse natural materials (e.g., processing waste RW) as novel substitutes for traditional activated carbon, as well as laccase as a green catalyst for the treatment of dye wastewater.

Adsorption of Cu2+, Pb2+, and Cd2+ onto oiltea shell from water

Agro-processing waste oiltea shell (OTS) has been previously proven effective in removing dye from water by adsorption. This study further evaluated its capability to adsorb heavy metals in aqueous solution. Adsorbent dosage, pH, ion concentration, temperature, and contact time were investigated in batch experiments. Pseudo-second order kinetic better described the adsorption process for the three ions. Langmuir equation was more suitable for Cu2+ and Cd2+, while Freundlich for Pb2+. The maximum adsorption capacity was 22.4 (Pb2+), 12.1 (Cu2+), and 14.2 mg/g (Cd2+), respectively. The adsorption was spontaneous and endothermic. Adsorption was then performed in column and Thomas model was fitted, based on which the maximum adsorption capacity was 7.42 (Cu2+), 4.17 (Pb2+), and 18.02 mg/g (Cd2+), respectively. High removal rate of metal ions and dye methylene blue was also achieved from their mixture solution. OTS is therefore a promising biosorbent in treating wastewater with organic and inorganic pollutants.