Structural elucidation and targeted valorization of untractable lignin from pre-hydrolysis liquor of xylose production via a simple and robust separation approach.

Effective separation of lignin macromolecules from the xylose pre-hydrolysates (XPH) during the xylose production, thus optimizing the separation and purification process of xylose, is of great significance for reducing the production costs, achieving the high value-added utilization of lignin and increasing the industrial revenue. In this study, a simple and robust method (pH adjustment) for the separation of lignin from XPH was proposed and systematically compared with the conventional acid-promoted lignin precipitation method. The results showed that the lignin removal ratio (up to 60.34 %) of this simple method was higher than that of the conventional method, and the proposed method eliminated the necessity of heating and specialized equipment, which greatly reduced the separation cost. Meanwhile, this simple method does not destroy the components in XPH (especially xylose), ensuring the yield of the target product. On the other hand, the obtained lignin was nano-scale with less condensed structures, which also possessed small molecular weights with narrow distribution, excellent antioxidant activity (8-14 times higher than commercial antioxidants) and UV protection properties. In conclusion, the proposed simple separation method could effectively separate lignin from XPH at low cost, and the obtained lignin had potential commercial applications, which would further enhance the overall profitability of industrial production.

Recovery Processes of Organic Acids from Fermentation Broths in the Biomass-Based Industry.

The new movement towards green chemistry and renewable feedstocks makes microbial production of chemicals more competitive. Among the numerous chemicals, organic acids are more attractive targets for process development efforts in the renewable-based biorefinery industry. However, most of the production costs in microbial processes are higher than that in chemical processes, among which over 60% are generated by separation processes. Therefore, the research of separation and purification processes is important for a promising biorefinery industry. This review highlights the progress of recovery processes in the separation and purification of organic acids, including their advantages and disadvantages, current situation, and future prospects in terms of recovery yields and industrial application.

Structurally related ganoderic acids induce apoptosis in human cervical cancer HeLa cells: Involvement of oxidative stress and antioxidant protective system.

Ganoderic acids (GAs) produced by Ganoderma lucidum possess anticancer activities with the generation of reactive oxygen species (ROS). However, the role of oxidative stress in apoptotic process induced by GAs is still undefined. In this study, the effects of four structurally related GAs, i.e. GA-T, GA-Mk, and two deacetylated derivatives of GA-T (GA-T1 and GA-T2) on the antioxidant defense system and induced apoptosis in cervical cancer cells HeLa were investigated in vitro. Our results indicated that the tested GAs (5-40 µM) induced apoptotic cell death through mitochondrial membrane potential decrease and activation of caspase-9 and caspase-3. Furthermore, GAs increased the generation of intracellular ROS and attenuated antioxidant defense system by decreasing glutathione (GSH) level, superoxide dismutase (SOD) and glutathione peroxidase (GPX) activities. The above effects were remarkably blocked by the exogenous antioxidants, i.e. N-acetylcysteine, catalase and diphenyleneiodonium chloride. The potency of the four GAs toward induced apoptosis, generation of ROS and suppression of antioxidant defense system was in the order of: GA-T > GA-Mk ≈ GA-T1 > GA-T2 in HeLa cells. These findings suggest that GAs induced mitochondria-dependent cell apoptosis in HeLa cells are mediated via enhancing oxidative stress and depressing antioxidant defense. Additionally, the acetylation of hydroxyl groups in GAs may contribute to their pro-oxidant activities and cytotoxicity, which is helpful to the development of novel chemotherapy agents.

[Study on the pH-sensitive secondary structure of gamma-PGA embedded with magnetite nanoparticles].

The conformational transition of poly gamma-glutamic acid (gamma-PGA) embedded with magnetite nanoparticles under various pH conditions was investigated by Fourier transform infrared spectroscopy (FTIR). The secondary structure content was determined through the analysis of amide I bands of Fourier deconvolution spectra, secondary derivative spectra and the Gaussian curve fitting of the original infrared spectra. The results showed that the conformation of the gamma-PGA was affected by solution pH. The total contents of beta-sheet and beta-turn were higher than 65%, while alpha-helix and random coil were low. The content of beta-turn increased with increasing pH, while the beta-sheet decreased. Additionally, the zeta potential results showed that the pH-sensitive secondary structure of gamma-PGA had influence on the stability of suspension of magnetic gamma-PGA nanospheres. The minimum value of zeta potential (-35. 4 mV) was obtained at pH 10.2.

In situ magnetic separation and immobilization of dibenzothiophene-desulfurizing bacteria.

In situ cell separation and immobilization of bacterial cells for biodesulfurization were developed by using superparamagnetic Fe(3)O(4) nanoparticles (NPs). The Fe(3)O(4) NPs were synthesized by coprecipitation followed by modification with ammonium oleate. The surface-modified NPs were monodispersed and the particle size was about 13 nm with 50.8 emu/g saturation magnetization. After adding the magnetic fluids to the culture broth, Rhodococcus erythropolis LSSE8-1 cells were immobilized by adsorption and then separated with an externally magnetic field. The maximum amount of cell mass adsorbed was about 530 g dry cell weight/g particles to LSSE8-1 cells. Analysis showed that the nanoparticles were strongly absorbed to the surface and coated the cells. Compared to free cells, the coated cells not only had the same desulfurizing activity but could also be easily separated from fermentation broth by magnetic force. Based on the adsorption isotherms and Zeta potential analysis, it was believed that oleate-modified Fe(3)O(4) NPs adsorbed bacterial cells mainly because of the nano-size effect and hydrophobic interaction.

Spontaneous vesicle formation of poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) triblock copolymer.

A novel method has been developed to prepare vesicles from aqueous solutions of poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) triblock copolymer, by adding anionic surfactant sodium dodecyl sulfate (SDS) and inorganic salt NaF. As determined by TEM and dynamic light scattering (DLS) measurements, the average diameter of vesicles is about 800 nm having 50 nm outer shell thickness. Identifying hydrophobic interactions between the block copolymers and the microenvironments around the vesicles using FTIR, 1H NMR, and fluorescence spectroscopy techniques revealed the vesicle formation mechanism. The spontaneously formed vesicles were further cross-linked by converting the terminal hydroxyl groups of block copolymers into aldehydes, and then chemically bridging the polymer chains by the reaction between aldehydes and diamine compounds. The cross-linked vesicles are proved much more stable than free vesicles even at higher dilutions. The obtained vesicles with good stability and biocompatibility are promising candidates for widespread applications.

Temperature-responsive magnetite/PEO-PPO-PEO block copolymer nanoparticles for controlled drug targeting delivery.

In this study, temperature-responsive magnetite/polymer nanoparticles were developed from iron oxide nanoparticles and poly(ethyleneimine)-modified poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) (PEO-PPO-PEO) block copolymer. The particles were characterized by TEM, XRD, DLS, VSM, FTIR, and TGA. A typical product has an approximately 20 nm magnetite core and an approximately 40 nm hydrodynamic diameter with a narrow size distribution and is superparamagnetic with large saturation magnetization (51.34 emu/g) at room temperature. The most attractive feature of the nanoparticles is their temperature-responsive volume-transition property. DLS results indicated that their average hydrodynamic diameter underwent a sharp decrease from 45 to 25 nm while evaluating the temperature from 20 to 35 degrees C. The temperature-dependent evolution of the C-O stretching band in the FTIR spectra of the aqueous nanoparticles solution revealed that thermo-induced self-assembly of the immobilized block copolymers occurred on the magnetite solid surfaces, which is accompanied by a conformational change from a fully extended state to a highly coiled state of the copolymer. Consequently, the copolymer shell could act as a temperature-controlled "gate" for the transit of guest substance. The uptake and release of both hydrophobic and hydrophilic model drugs were well controlled by switching the transient opening and closing of the polymer shell at different temperatures. A sustained release of about 3 days was achieved in simulated human body conditions. In primary mouse experiments, drug-entrapped magnetic nanoparticles showed good biocompatibility and effective therapy for spinal cord damage. Such intelligent magnetic nanoparticles are attractive candidates for widespread biomedical applications, particularly in controlled drug-targeting delivery.

Probing paeonol-pluronic polymer interactions by 1H NMR spectroscopy.

By using a combination of 1H NMR spectroscopy, two-dimensional heteronuclear single-quantum coherence-resolved (1)H{(13)C} and homonuclear rotating-frame Overhauser enhancement NMR correlation experiments with diffusion ordered spectroscopy (DOSY), the location and distribution of a hydrophobic drug, paeonol, have been established with respect to the methyl groups of the poly(ethylene oxide)-poly(propylene oxide) -poly(ethylene oxide) (PEO-PPO-PEO) triblock copolymer. The interaction between them is adjustable according to the different temperature-dependent hydrophilicities or hydrophobicities of the triblock copolymer components. On the other hand, such interactions influence the self-assembly properties of the block copolymer amphiphiles in solution. The amount of anhydrous methyl groups of PPO segments shows an increase with increasing paeonol concentration. It was also demonstrated that the shell-crosslinking of the Pluronic polymer has an effect in increasing the amount of anhydrous methyl groups and thus increasing the hydrophobicity of Pluronic micelles. This might be the deeper reason underlying the increase in drug-loading capacity and prolongation in release time of Pluronic micelles for drug delivery after the shell-crosslinking. Changes in self-diffusion coefficients of paeonol with varying copolymer concentrations and types were also determined by the diffusion-based NMR DOSY technique, and values of K(a), DeltaG, and n were calculated.

Oil-induced aggregation of block copolymer in aqueous solution.

The oil-induced aggregation behavior of PEO-PPO-PEO Pluronic P84 [(EO)19(PO)39(EO)19] in aqueous solutions has been systematically investigated by 1H NMR spectroscopy, freeze-fracture transmission electron microscopy (FF-TEM), and dynamic light scattering (DLS). The critical micellization temperature (CMT) for P84 in the presence of oils decreases with increasing oil concentration. The effectiveness of various oils in decreasing the CMT of block copolymer follows the order m-xylene (C(8)H(10)) > toluene (C(7)H(8)) > benzene (C(6)H(6)) > n-octane (C(8)H(18)) > n-hexane (C(6)H(14)) approximately cyclohexane (C(6)H(12)). It was found that the amount of anhydrous PO methyl groups increases whereas the amount of hydrated PO methyl groups decreases upon the addition of oils. At low oil concentration, the oil molecules are entrapped by the micellar core, but as the oil concentration increases above a certain value, the micellar core swells significantly as a result of the penetrated oil molecules, and much larger aggregates are formed. Intermolecular rotating-frame nuclear Overhauser effect (ROE) measurements between P84 and benzene were performed at 10 and 40 degrees C. The specific interaction between benzene and the methyl groups of PPO was determined, and it was observed that the interaction site remained unchanged as the temperature was increased.

1H NMR spectroscopic investigations on the micellization and gelation of PEO-PPO-PEO block copolymers in aqueous solutions.

The effects of temperature, polymer composition, and concentration on the micellization and gelation properties of poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) (PEO-PPO-PEO) block copolymers in aqueous solutions were investigated by 1H NMR spectroscopy. It was found that the temperature-dependent behavior of PPO blocks, observed as changes in chemical shift, half-height width, and integral value, could be attributed as an intrinsic tool to characterize the transition states during unimer to micelle formation. The 1H NMR spectral analysis revealed that the hydrophobic part, PPO, of the Pluronic polymers plays a more significant role in the temperature-induced micellization, whereas the transitional behavior of Pluronic polymer, i.e., from micellization to liquid crystals formation, resulted in the drastic broadening of the spectral signals for the PEO, indicating that the PEO segments play a more significant role in the crystallization process. It was also observed that the temperature-dependent changes in the half-height width of the PEO -CH2- signal are sensitive to the liquid crystalline phase formation, which could be attributed to the close packing of spherical micelles at high polymer concentrations or temperatures.

Interaction of urea with pluronic block copolymers by 1H NMR spectroscopy.

Solution 1H NMR techniques were used to characterize the interaction of urea with poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) (PEO-PPO-PEO) triblock copolymers. The urea was established to interact selectively with the PEO blocks of the block copolymer, and the interaction sites were found not to change with increasing temperature. Such interactions influence the self-assembly properties of the block copolymer in solution by increasing the hydration of the block copolymers and stabilizing the gauche conformation of the PPO chain. Therefore, urea increases the critical micellization temperature (CMT) values of PEO-PPO-PEO copolymers, and the effect of urea on the CMT is more pronounced for copolymers with higher PEO contents and lower for those with increased contents of PPO segments.

Salt-induced micellization of a triblock copolymer in aqueous solution: a 1H nuclear magnetic resonance spectroscopy study.

An aqueous micellar solution of a PEO-PPO-PEO triblock copolymer, pluronic F88 (EO103PO39EO103), in the presence of salt (KCl) has been investigated by 1H NMR spectroscopy. The hydrogen-bonding structure in water is directly changed by the strong polarization effect of added salt, which indirectly weakens the interaction of polymer molecules with water. Both EO and PO blocks are dehydrated by the addition of salt in a similar way, whereas the solubility of the PO blocks may be affected in a more pronounced way, which results in the decrease of the critical micellization temperature (CMT). It is found that the addition of salt favors a more compact micellar core, where the water content is decreased and an effective PO-PO interaction is increased. Increasing the salt concentration would result in a decrease in the number of gauche conformers in the PPO chain, which may be the deeper reason for the decreasing solubility of PPO segments in aqueous salt solution. The temperature region over which the micellization occurs is broad, indicating that micelles and unimers coexist over an extended temperature range, whereas this transition region is significantly narrowed by the addition of salt. The addition of salt offers a good substitute way of changing the temperature to induce micellization. The critical micellization salt concentration (CMSC) is determined to be 1.0 mol l-1 for KCl in 2.5% aqueous pluronic F88 solution at 25 degrees C, and the transition region in which both free and associated copolymer molecules coexist is defined to range from 1 to 2 mol L-1.

[Study on interaction of anionic surfactant SDS and bovine serum albumin by fourier transform infrared spectroscopy].

FTIR spectroscopy was applied to investigate the interaction of anionic surfactant Sodium Dodecyl Sulfate (SDS) and Bovine Serum Albumin (BSA). Amide band I of BSA was analyzed to obtain the change in secondary structure of BSA when different concentration of SDS was added and during different interaction period. In short interaction period and at low concentration of SDS, the alpha-helixes increased and the random coil decreased. In long interaction period or at high concentration of SDS, SDS unfolded the protein by decreasing the alpha-helix structure and increasing the random coil.

Effect of hydrophobicity inside PEO-PPO-PEO block copolymer micelles on the stabilization of gold nanoparticles: experiments.

In this paper we present the effect of poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) block copolymer micelles and their hydrophobicity on the stabilization of gold nanoparticles. Gold nanoparticles were prepared by a method developed by Sakai et al. (Sakai, T.; Alexandridis, P. Langmuir 2004, 20, 8426). An absorption centered at 300-400 nm in time-dependent UV spectra provided evidence that the very first step of the synthesis was to form primary gold clusters. Then the gold clusters grew in size and were stabilized by block copolymer micelles. The stabilization capacities of the micelles were modulated by tuning the block copolymer concentration and composition and by adding salts. With good stabilization, gold particles were spherical and uniform in size with a diameter of 5-10 nm. Otherwise they were aggregates with irregular shapes such as triangular, hexagonal, and rodlike. The presence of a small amount of NaF significantly increased the stabilization capacity of the micelles and consequently modified the quality of the gold particles. Using FTIR and 1H NMR spectroscopy, micellization of the block copolymers and hydrophobicity of the micelles were proven very important for the stabilization. A higher hydrophobicity of the micelle cores was expected to favor the entrapment of primary gold clusters and the stabilization of gold nanoparticles.

Purification of nattokinase by reverse micelles extraction from fermentation broth: effect of temperature and phase volume ratio.

Nattokinase is a novel fibrinolytic enzyme that is considered to be a promising agent for thrombosis therapy. In this study, reverse micelles extraction was applied to purify and concentrate nattokinase from fermentation broth. The effects of temperature and phase volume ratio used for the forward and backward extraction on the extraction process were examined. The optimal temperature for forward and backward extraction were 25 degrees C and 35 degrees C respectively. Nattokinase became more thermosensitive during reverse micelles extraction. And it could be enriched in the stripping phase eight times during backward extraction. It was found that nattokinase could be purified by AOT reverse micelles with up to 80% activity recovery and with a purification factor of 3.9.

[Comparison of the desulfurization activity among several bacteria and analysis of the conservation of their desulfurization genes].

Several bacteria, Bacillus brevis R-6, Pseudomonas delafleldii R-8, Nocardia globerula R-9, Bacillus sphaericus R-16, Rhodococcus erythropolis LSSE8-1 and Gordonia nitida LSSEJ-1, which can convert dibenzothiophene into 2-hydroxybiphenyl and sulfate, were investigated. Desulfurization products were quantitively determined by HPLC. Result revealed that each of these bacteria desulfurize DBT at a different rate. In order to obtain more information, the fragments encoding desulfurizing enzymes were studied. Desulfurization genes of R-6 and R-8 were separately amplified via PCR with specific primers based on the related sequences of Rhodococcus sp. IGTS8. Both sequences areminimally 99% related to IGTS8 sequence. Afterwards, dsz operon of LSSEJ-1 and R-9 were amplified and cloned. Sequences are also highly conservative. Data shows that identity of dszA between R-9 and IGTS8 is 99.6%, and identity of dszA between LSSEJ-1 and IGTS8 is 99.9%; dszB sequence of R-9 and LSSEJ-1 is 99.6% similarity to their counterpart sequence from IGTS8;Identity of dszC between R-9 and IGTS8 is 99.9%, and identity of dszC between LSSEJ-1 and IGTS8 is 99.1% . It may be deduced that the origins of desulfurization genes from mesophilic bacteria are the same.

Effect of sodium chloride on association behavior of poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) block copolymer in aqueous solutions.

The effect of sodium chloride (NaCl) upon the thermally induced association behavior of poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) (PEO-PPO-PEO) block copolymer, Pluronic P103, has been investigated using pyrene fluorescence spectroscopy. The critical micellization temperature (CMT) of Pluronic P103 in aqueous solution is decreased by the addition of NaCl. The standard enthalpy and entropy of micellization for Pluronic P103 in water are increased in the presence of small amounts of NaCl, but further addition of NaCl decreases the standard enthalpy and entropy of micellization. The I1/I3 ratio of pyrene in aqueous Pluronic P103 solutions at temperature below the CMT decreases with increases of NaCl concentration, which is related to the decrease of PPO solubility. The decrease in polarity of the PPO shifts the CMT toward lower temperature.

Immobilization of Pseudomonas delafieldii with magnetic polyvinyl alcohol beads and its application in biodesulfurization.

Pseudomonas delafieldii was immobilized in magnetic polyvinyl alcohol (PVA) beads using a hydrophilic magnetic fluid, which was prepared by a co-precipitation method. The beads had distinct super-paramagnetic properties and were compared with immobilized cells in non-magnetic PVA beads. Their desulfurizing activity was increased slightly from 8.7 to 9 mmol sulfur kg(-1) (dry cell) h(-1). The main advantages was that the magnetic immobilized cells maintain a high desulfurization activity and remain in good shape after 7 times of repeated use, while the non-magnetic immobilized cells could only be used for 5 times. Furthermore, the magnetic immobilized cells could be easily collected or separated magnetically from the biodesulfurization reactor.

Melt, hydration, and micellization of the PEO-PPO-PEO block copolymer studied by FTIR spectroscopy.

Fourier transform infrared (FTIR) spectroscopy was used to study the conformational changes of the polyethylene oxide-polypropylene oxide-polyethylene oxide (PEO-PPO-PEO) block copolymer, Pluronic P104, in a large concentration range in a polymer-water system as a function of temperature. The melt in which the conformational transition of the PEO blocks occurs gives remarkable changes in the spectral behavior. A small amount of water in Pluronic P104 can induce the PEO block amorphism. The addition of more water only swells the PEO dominant region and gives no significant difference in the conformational structure of the block copolymer in the ordered phases of Pluronic P104-water mixtures. The PPO blocks of Pluronic P104 are hydrated only in a condition of lower temperature and higher water content. The temperature dependent micellization of Pluronic P104 in water was analyzed by a FTIR spectroscopic method. The appearance of the symmetric deformation band of the anhydrous methyl groups at temperature below the CMT indicates the existence of a hydrophobic microenvironment. The appearance of the symmetric deformation band of the hydrated methyl groups at higher temperatures indicates that the micellar core must contain some amount of water. The results of FTIR data show that the proportion of the anhydrous methyl groups increases and water content in the micellar core decreases during the micellization process.