The impact of adsorbed cellulase inactivation on enzymatic hydrolysis kinetics.

Several technical and economic obstacles currently hamper the industrial development of ethanol from biomass. One of the key bottlenecks is the slow kinetics of the enzymatic hydrolysis of cellulose, and the subsequent rate reduction as the reaction proceeds. As a result, this research focused on understanding underlying causes for the slow kinetics, rate reduction, and low yield during cellulose hydrolysis. Mechanisms traditionally thought to cause these results were investigated, such as change of substrate properties and deactivation of enzyme due to environmental mechanisms, but neither was found to contribute significantly to the slow kinetics and low yield. Inactivation due to enzyme-substrate interactions was then proposed as a key factor. Results here show that inactivation of adsorbed enzyme played the most significant role for the hydrolysis rate reduction and low yield based on the following findings: (1) a kinetic model featuring inactivation of adsorbed enzyme accurately accounted for experimental cellulose hydrolysis data for two different types of substrates; the enzyme\u27s apparent maximum reaction rate was found to decrease with a first order exponential decay function of time due to inactivation of the adsorbed enzyme, which has historically always been considered to remain constant. (2) comparison of relative extents of enzyme activity loss due to environmental mechanisms (such as thermal and/or mechanical factors) with inactivation due to enzyme-substrate interactions revealed that enzyme- substrate interactions contributed more towards the overall activity loss than did environmental mechanisms; (3) AFM imaging visualized crowding of Cellobiohydrolase 1 (CBHl) on cellulose substrate surface and thereafter became inactivated; (4) desorption of inactive CBHl was slower compared to desorption of active CBHl, implying that once inactivated, CBH 1 cannot dissociate immediately to find another site on a substrate surface to start another digestive cycle. The overall conclusion is that inactivation of adsorbed enzyme is a primary contributor to the hydrolysis rate reduction. Near complete conversion (99%) of cellulose was predicted by the model to occur within 10-20 hours if inactivation of adsorbed cellulase can be prevented, compared to 7-10 days or more to achieve a lower yield when inactivation occurs. Finally, factors to consider when developing a cellulose hydrolysis process were proposed based on the inactivation mechanism. One important strategy proposed is to desorb inactive cellulases from the substrate, such as with the addition of GdnHCl. Additionally, a technique for scaling-up separation of CBHl was developed. The technique allows for efficient purification of active CBHl from commercial cellulose cocktails at a cost of less than 10% compared to the conventional small-scale FPLC method

Enhanced phosphate removal under an electric field via multiple mechanisms on MgAl-LDHs/AC composite electrode

Phosphorus removal is essential to avoid eutrophication in water bodies. Layered double hydroxides (LDHs) are widely used to scavenge phosphate through intercalated ion exchange or surface complexation. Moreover, LDHs have attracted increasing attention as electrode modifiers for supercapacitors. Researchers have begun to re-delve the electrosorption technology according to the fundamental principle of electrical double layers. Herein, we propose a new phosphate removal method inspired by the adsorption characteristic and electrical double-layer capacitive properties of LDHs through electrosorption via capacitive deionization. We present a series of experiments to study the enhanced phosphate removal under an electric field via multiple mechanisms on the MgAl-LDHs/AC electrode. The uptake of phosphate by MgAl-LDHs/AC was investigated as a function of phosphate concentration, applied voltage, electrode materials, and temperature. The MgAl-LDHs/AC electrode possessed a salt removal capacity of 67.92 mg PO43−·g−1 (1.2 V, 250 mg·L−1 KH2PO4, 30 °C). The electrosorption kinetics of phosphate ions onto the capacitive deionization electrode followed the pseudo-second-order kinetics model rather than the pseudo-first-order kinetics model. Furthermore, the adsorption isotherms of phosphate on the MgAl-LDHs/AC electrode fitted the Freundlich model better than the Langmuir model. The proposed method could be used for phosphate removal

Influence of various experimental parameters on the capacitive removal of phosphate from aqueous solutions using LDHs/AC composite electrodes

The efficient uptake of phosphate from aqueous solutions was achieved on layered double hydroxides (LDHs)-based electrodes via capacitive desalination in our previous study. The current follow-up work was mainly carried out to study the influence of various experimental parameters on the capacitive removal of phosphate using LDHs/activated carbon (LDHs/AC) composite electrodes. A series of batch experiments were implemented to investigate the experimental factors, including Mg2+/Al3+ ratios (2, 3, and 4), trivalent metal cations (Al3+, Fe3+, Cr3+), initial solution pH (from 3 to 10), coexisting anions (NO3-, Cl-, SO42-), and ion strengths, in capacitive deionization. The electrode materials before and after capacitive deionization were characterized to reinforce the analysis of the adsorption mechanisms by X-ray powder diffraction, scanning electron microscopy, energy dispersive X-ray, cyclic voltammetry, and electrochemical impedance spectroscopy. Results indicated that the Mg-Al LDHs/AC electrodes exhibited higher phosphate adsorption capacity (80.43 mg PO43-/g), more regular morphology, and higher degree of crystallinity than that of Mg-Fe LDHs/AC and Mg-Cr LDHs/AC. Increasing Mg2+/Al3+ ratios enhanced the adsorption capacity of phosphate. The uptake of phosphate by Mg-Al LDHs/AC under circumneutral pH and low ion strength reached the maximum level. Furthermore, the presence of coexisting anions lowered the adsorption capacity of phosphate mainly due to the occurrence of a compressed electrical double layer. Therefore, the influence of different experimental parameters on phosphate removal via capacitive deionization by Mg-Al LDHs/AC necessitates a systematic investigation to optimize the preparation conditions of LDHs-based electrodes and several important operating parameters

Enhanced hydrophilicity and antibacterial activity of PVDF ultrafiltration membrane using Ag3PO4/TiO2 nanocomposite against E. coli

Ag3PO4/TiO2, nanocomposite was fabricated by an in situ precipitation method and then blended into poly(vinylidene fluoride) (PVDF) casting solution to prepare the ultrafiltration membrane via wet phase inversion technique. The water flux and bovine serum albumin (BSA) rejection rate of membrane were investigated; meanwhile, the ultrafiltration membrane morphologies and structural properties were analyzed using scanning electron microscope (SEM) and X-ray diffraction. Compared with the control membrane, the permeate performance of blended membranes was improved while possessing a steady BSA retention due to enhanced hydrophilicity. Mechanical tests revealed that the modified membranes exhibited a larger tensile strength and breakage elongation. SEM images and the halo zone testing were employed to assess the antibacterial performances of the nanocomposite membranes against Escherichia coli. The antibacterial tests confirmed that the modified membranes showed an effective antibacterial property against E. coli

Longan seed and mangosteen skin based activated carbons for the removal of Pb(II) ions and rhodamine-B dye from aqueous solutions

Agricultural biomass wastes of longan seed and mangosteen skin were collected as precursors to prepare activated carbons (LS-AC-5 and MS-AC-5, respectively) through carbonization at medium temperature and KOH activation at high temperature. Their pore structures, structural properties and surface morphologies were characterized by X-ray diffractometer, Brunauer–Emmett–Teller surface measurement system, and scanning electron microscopy, respectively. Effects of contact time and pH on adsorption performances of samples were investigated by removal of Pb(II) ions and Rhodamine-b from aqueous solutions. Experimental adsorption isotherms of Rhodamine-b and Pb(II) ions on LS-AC-5 and MS-AC-5 fitted well with the Langmuir model. Results further showed that MS-AC-5 exhibited a larger surface area of 2960.56 m2/g and larger portions of micropores and mesopores (pore volume of 1.77 cm3/g) than LS-AC-5 (surface area: 2728.98 m2/g; pore volume: 1.39 cm3/g). Maximum monolayer adsorption capabilities of 1265.82 and 117.65 mg/g for Rhodamine-b and Pb(II) ions on MS-AC-5 were higher than those on LS-AC-5 (1000.20 and 107.53 mg/g), respectively

Sustainable composite super absorbents made from polysaccharides

Compared to traditional super absorbent polymers using raw materials from petrochemical industry, natural polymer absorbents are more favorable because they are sustainable and biodegradable. In this study, composite absorbents were developed by crosslinking carrageenan with sodium alginate using calcium chloride. Effect of composition on absorption was tested. Absorption was improved by increasing carrageenan content. The super absorbent exhibited the maximal swelling ratio of 13.1 g/g in 0.9% saline water in just 5 min. The maximal tensile strength was reached with a value of 12.8 MPa. Water contact angle revealed that carrageenan is more hydrophobic than sodium alginate. Presence of sulfate groups might be a key factor promoting absorption. The scanning electron microscopic images showed that the composite material had a structure with alginate arranged at the outside surface. These results demonstrate that a sustainable and biodegradable absorbent was successfully developed with a matrix of properties for potential application in diapers

Sulfonated Binaphthyl-Containing Poly(arylene ether ketone)s with Rigid Backbone and Excellent Film-Forming Capability for Proton Exchange Membranes

Sterically hindered (S)-1,1′-binaphthyl-2,2′-diol had been successfully copolymerized with 4,4′-sulfonyldiphenol and 4,4′-difluorobenzophenone to yield fibrous poly(arylene ether ketone)s (PAEKs) containing various amounts of binaphthyl unit, which was then selectively and efficiently sulfonated using ClSO3H to yield sulfonated poly(arylene ether ketone)s (SPAEKs) with ion exchange capacities (IECs) ranging from 1.40 to 1.89 mmol·g−1. The chemical structures of the polymers were confirmed by 2D 1H⁻1H COSY NMR and FT-IR. The thermal properties, water uptake, swelling ratio, proton conductivity, oxidative stability and mechanical properties of SPAEKs were investigated in detail. It was found that the conjugated but non-coplanar structure of binaphthyl unit endorsed excellent solubility and film-forming capability to SPAEKs. The SPAEK-50 with an IEC of 1.89 mmol·g−1 exhibited a proton conductivity of 102 mS·cm−1 at 30 °C, much higher than that of the state-of-the-art Nafion N212 membrane and those of many previously reported aromatic analogs, which may be attributed to the likely large intrinsic free volume of SPAEKs created by the highly twisted chain structures and the desirable microscopic morphology. Along with the remarkable water affinity, thermal stabilities and mechanical properties, the SPAEKs were demonstrated to be promising proton exchange membrane (PEM) candidates for potential membrane separations

Alginate Adsorbent Immobilization Technique Promotes Biobutanol Production by Clostridium acetobutylicum Under Extreme Condition of High Concentration of Organic Solvent

In Acetone-Butanol-Ethanol fermentation, bacteria should tolerate high concentrations of solvent products, which inhibit bacteria growth and limit further increase of solvents to more than 20 g/L. Moreover, this limited solvent concentration significantly increases the cost of solvent separation through traditional approaches. In this study, alginate adsorbent immobilization technique was successfully developed to assist in situ extraction using octanol which is effective in extracting butanol but presents strong toxic effect to bacteria. The adsorbent improved solvent tolerance of Clostridium acetobutylicum under extreme condition of high concentration of organic solvent. Using the developed technique, more than 42% of added bacteria can be adsorbed to the adsorbent. Surface area of the adsorbent was more than 10 times greater than sodium alginate. Scanning electron microscope image shows that an abundant amount of pore structure was successfully developed on adsorbents, promoting bacteria adsorption. In adsorbent assisted ABE fermentation, there was 21.64 g/L butanol in extracting layer compared to negligible butanol produced with only the extractant but without the adsorbent, for the reason that adsorbent can reduce damaging exposure of C. acetobutylicum to octanol. The strategy can improve total butanol production with respect to traditional culture approach by more than 2.5 fold and save energy for subsequent butanol recovery, which effects can potentially make the biobutanol production more economically practical

Comparative Pathogenicity and Transmissibility of the H7N9 Highly Pathogenic Avian Influenza Virus and the H7N9 Low Pathogenic Avian Influenza Virus in Chickens

There were five outbreaks of H7N9 influenza virus in humans in China since it emerged in 2013, infecting >1000 people. The H7N9 low pathogenic influenza virus was inserted into four amino acids in the HA protein cleavage site to mutate into the H7N9 highly pathogenic virus. This emerging virus caused 15 outbreaks in chickens from the end of 2016 to date. Two H7N9 avian influenza virus (AIV) strains, A/chicken/Guangdong/A46/2013 (LPAIV) and A/chicken/Guangdong/Q29/2017 (HPAIV), were selected to compare the pathogenicity and transmissibility between H7N9 LPAIVs and HPAIVs in chickens. We inoculated 3- to 4-week-old specific-pathogen-free (SPF) chickens with 6 log10EID50/0.1 mL viruses via the ocular-nasal route and co-housed four chickens in each group. The inoculated chicken mortality rate in the A46 and Q29 groups was 1/5 and 5/5, respectively. Q29 virus replication was more efficient compared to the A46 virus in inoculated chickens. Infected chickens initiated viral shedding to naïve contact chickens through respiratory and digestive routes. Both viruses transmitted between chickens by naïve contact, but the Q29 virus had a higher pathogenicity in contact chickens than the A46 virus. Compared with early H7N9 LPAIVs, the pathogenicity and transmissibility of the emerging H7N9 HPAIV was stronger in chickens, indicating that H7N9 influenza virus may continue to threaten human and poultry health

Second fractionation (cation exchange) of fractions 7–10 from the separation shown in Figure 1.

<p><b>(A)</b> OD₂₈₀. The red box indicates the fractions collected for the third-step separation. <b>(B)</b> Activity with (black solid line) or without (red dashed line) 8-CSM as described in the legend to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0109885#pone-0109885-g001" target="_blank">Fig. 1</a>. Glc released by 8-CSM alone was 0.24±0.01 g/L. <b>(C)</b> Degree of synergy calculated from (B).</p