Determining intracellular lipid content of different oleaginous yeasts by one simple and accurate Nile Red fluorescent method.

A simple and accurate Nile Red fluorescent method was built to evaluate the lipid content of three different oleaginous yeasts by one standard curve. The staining of cells can be observed clearly by laser scanning confocal microscope, showing that Nile Red can enter into the cells of oleaginous yeasts easily. A series of conditions such as pretreating temperature, cell suspension concentration (OD600), staining time, Nile Red concentration and the type of suspension solvent were learnt systematically to obtain the optimal process parameters for Nile Red staining. After optimization, the fitting curve of Nile Red fluorescent method was established under suitable conditions (pretreating temperature: 50 °C, OD600: 1.0; staining time: 5 mins; Nile Red concentration: 1.0 µg/mL; suspension solvent: PBS) and it had a suitable correlation coefficient (R2 = 0.95) for lipid content measurement of different oleaginous yeasts. By this study, the possibility of lipid content determination of different oleaginous yeasts by one fitting curve can be proven and this will improve the efficiency of researches related to microbial lipid production.

Effect of propofol and sevoflurane on cognitive function among elderly patients undergoing elective surgery under anesthesia.

The objective of present study was to compare the effect of propofol and sevoflurane on cognitive function among elderly patients undergoing elective surgery under anesthesia. Elderly patient who met eligibility criteria were randomized to receive Intravenous anesthetic (propofol) or Inhalation anesthetic (sevoflurane, Group II) in allocation ratio of 1:1. The following variables were assessed, 1) recovery time; 2) measurement of attention and psychomotor functions; 3) memory (verbal memory); 4) obvious memories during anesthesia assessed by remembrance (recall) and recognition tests. A total of 200 patients were completed study. Statistical analysis showed that the recovery time was significantly greater in patients who received sevoflurane when compared to patients who received propofol (p<0.005). Patients who had treated with sevoflurane had greater reaction time compared to the patients who had been treated with propofol after 30- minutes of anesthesia (immediate test) [p<0.005]. Similar trend of results was observed between both the groups after 120 minutes of anesthesia (delayed test). Moreover, the patients who were treated with propofol had better memory score as compared to patients treated with sevoflurane. The difference was statistically significant between both the treatment groups in both type of recognition test (immediate and delayed recognition test) [p<0.005].

Lumping kinetics of ABE fermentation wastewater treatment by oleaginous yeast Trichosporon cutaneum.

Lumping kinetics models were built for the biological treatment of acetone-butanol-ethanol (ABE) fermentation wastewater by oleaginous yeast Trichosporon cutaneum with different fermentation temperatures. Compared with high temperature (33°C, 306 K) and low temperature (23°C, 296 K), medium temperature (28°C, 301 K) was beneficial for the cell growth and chemical oxygen demand (COD) degradation during the early stage of fermentation but the final yeast biomass and COD removal were influenced little. By lumping method, the materials in the bioconversion network were divided into five lumps (COD, lipid, polysaccharide, other intracellular products, other extracellular products), and the nine rate constants (k1-k9) for the models can well explain the bioconversion laws. The Gibbs free energy (G) for this bioconversion was positive, showing that it cannot happen spontaneous, but the existence of yeast can after the chemical equilibrium and make the bioconversion to be possible. Overall, the possibility of using lumping kinetics for elucidating the laws of materials conversion in the biological treatment of ABE fermentation wastewater by T. cutaneum has been initially proved and this method has great potential for further application.

Efficient bioconversion from acid hydrolysate of waste oleaginous yeast biomass after microbial oil extraction to bacterial cellulose by Komagataeibacter xylinus.

Biomass acid hydrolysate of oleaginous yeast Trichosporon cutaneum after microbial oil extraction was applied as substrate for bacterial cellulose (BC) production by Komagataeibacter xylinus (also named as Gluconacetobacter xylinus previously) for the first time. BC was synthesized in static culture for 10 days, and the maximum BC yield (2.9 g/L) was got at the 4th day of fermentation. Most carbon sources in the substrate (glucose, mannose, formic acid, acetic acid) can be utilized by K. xylinus. The highest chemical oxygen demand (COD) removal (40.7 ± 3.0%) was obtained at the 6th day of fermentation, and then the COD increased possibly due to the degradation of BC. The highest BC yield on COD consumption was 38.7 ± 4.0% (w/w), suggesting that this is one efficient bioconversion for BC production. The BC structure was affected little by the substrate by comparison with that generated in classical HS medium using field-emission scanning electron microscope (FE-SEM), Fourier transform infrared, and X-ray diffraction. Overall, this technology can both solve the issue of waste oleaginous yeast biomass and produce valuable biopolymer (BC).

Efficient Using Durian Shell Hydrolysate as Low-Cost Substrate for Bacterial Cellulose Production by Gluconacetobacter xylinus.

Durian is one important tropical fruit with high nutritional value, but its shell is usually useless and considered as waste. To explore the efficient and high-value utilization of this agricultural and food waste, in this study, durian shell was simply hydrolyzed by dilute sulfuric acid, and the durian shell hydrolysate after detoxification was used for bacterial cellulose (BC) production by Gluconacetobacter xylinus for the first time. BC was synthesized in static culture for 10 days and the highest BC yield (2.67 g/L) was obtained at the 8th day. The typical carbon sources in the substrate including glucose, xylose, formic acid, acetic acid, etc. can be utilized by G. xylinus. The highest chemical oxygen demand (COD) removal (16.40%) was obtained at the 8th day. The highest BC yield on COD consumption and the highest BC yield on sugar consumption were 93.51% and 22.98% (w/w), respectively, suggesting this is one efficient bioconversion for BC production. Durian shell hydrolysate showed small influence on the BC structure by comparison with the structure of BC generated in traditional Hestrin-Schramm medium detected by FE-SEM, FTIR, and XRD. Overall, this technology can both solve the issue of waste durian shell and produce valuable bio-polymer (BC).

Use of elephant grass (Pennisetum purpureum) acid hydrolysate for microbial oil production by Trichosporon cutaneum.

Elephant grass (Pennisetum purpureum) dilute acid hydrolysate contains 34.6 g/L total sugars. The potential of lipid production by oleaginous yeast Trichosporon cutaneum grown on elephant grass acid hydrolysate was investigated for the first time. During the fermentation process on the elephant grass acid hydrolysate, glucose, xylose, and arabinose could be well utilized as carbon sources by T. cutaneum. Interestingly, xylose was almost no use before glucose was consumed completely. This illustrated that simultaneous saccharification of xylose and glucose by T. cutaneum did not occur on elephant grass acid hydrolysate. The highest biomass, lipid content, lipid yield, and lipid coefficient of T. cutaneum were measured after the sixth day of fermentation and were 22.76 g/L, 24.0%, 5.46 g/L, and 16.1%, respectively. Therefore, elephant grass is a promising raw material for microbial oil production by T. cutaneum.

Bacterial cellulose production from the litchi extract by Gluconacetobacter xylinus.

Although litchi has both nutrient and edible value, the extremely short preservation time limited its further market promotion. To explore processed litchi products with longer preservation time, litchi extract was selected as an alternative feedstock for production of bacterial cellulose (BC). After 2 weeks of static fermentation, 2.53 g/L of the BC membrane was obtained. The trace elements including magnesium (Mg) and sodium (Na) in the litchi extract were partly absorbed in the BC membrane, but no potassium (K) element was detected in it, curiously. Scanning electron microscope (SEM) photographs exhibited an ultrafine network nanostructure for the BC produced in the litchi extract. Analysis of the fourier-transform infrared spectroscopy (FTIR) confirmed the pellicles to be a cellulosic material. Interestingly, X-ray diffraction (XRD) results showed the BC membrane obtained from litchi extract had higher crystallinity of 94.0% than that from HS medium. Overall, the work showed the potential of producing high value-added polymer from litchi resources.

Preparation of Bacterial Cellulose/Inorganic Gel of Bentonite Composite by In Situ Modification.

To evaluate the possibility of Bacterial cellulose/Inorganic Gel of Bentonite (BC/IGB) composite production using in situ method, the BC/IGB composite was successfully produced by in situ modification of BC in both HS medium and corncob hydrolysate. The results showed that the BC/IGB composite obtained in HS medium (one classical medium for BC production) had a higher water holding capacity, but the water retention capacity of the BC/IGB composite obtained in corncob hydrolysate was better. The performance of BC/IGB composite depended on the environment of in situ modification. Using different media showed significant influence on the sugar utilization and BC yield. In addition, BC/IGB composite produced by in situ method was compared with that produced by ex situ method, and the results shows that water holding capacity of BC/IGB composite obtained through in situ method was better. XRD results showed the crystallinity of BC/IGB composite related little to its performance as water absorbent. Overall, in situ modification is appropriate for further production of BC composite and other clay materials.

Beneficial effect of corncob acid hydrolysate on the lipid production by oleaginous yeast Trichosporon dermatis.

In this work, corncob acid hydrolysate and its simulated medium whose sugar composition was the same as the corncob acid hydrolysate were used as fermentation substrate for lipid production by oleaginous yeast Trichosporon dermatis. On the corncob acid hydrolysate, after 7 days of fermentation, the biomass, lipid content, lipid yield, and lipid coefficient of T. dermatis were 17.3 g/L, 40.2%, 7.0 g/L, and 16.5%, respectively. Interestingly, during the lipid fermentation on the corncob acid hydrolysate, glucose, xylose, arabinose, and even acetic acid could be well utilized as carbon sources by T. dermatis. Surprisingly, the lipid yield (7.0 g/L) of T. dermatis on the corncob acid hydrolysate was much higher than that (3.8 g/L) on the simulated medium, in spite of the fact that the lipid coefficient (17.4%) on the simulated medium was a little higher. This phenomenon further showed that lignocellulosic acid hydrolysate was a suitable substrate for lipid fermentation by T. dermatis. This work would help the comprehensive utilization of lignocellulosic biomass for lipid production.

Beneficial Effect of Acetic Acid on the Xylose Utilization and Bacterial Cellulose Production by Gluconacetobacter xylinus.

In this work, acetic acid was found as one promising substrate to improve xylose utilization by Gluconacetobacter xylinus CH001. Also, with the help of adding acetic acid into medium, the bacterial cellulose (BC) production by G. xylinus was increased significantly. In the medium containing 3 g l(-1) acetic acid, the optimal xylose concentration for BC production was 20 g l(-1). In the medium containing 20 g l(-1) xylose, the xylose utilization and BC production by G. xylinus were stimulated by acetic acid within certain concentration. The highest BC yield (1.35 ± 0.06 g l(-1)) was obtained in the medium containing 20 g l(-1) xylose and 3 g l(-1) acetic acid after 14 days. This value was 6.17-fold higher than the yield (0.21 ± 0.01 g l(-1)) in the medium only containing 20 g l(-1) xylose. The results analyzed by FE-SEM, FTIR, and XRD showed that acetic acid affected little on the microscopic morphology and physicochemical characteristics of BC. Base on the phenomenon observed, lignocellulosic acid hydrolysates (xylose and acetic acid are main carbon sources present in it) could be considered as one potential substrate for BC production.

Dose-response of epidural ropivacaine with 0.4†µg†mL-1 of dexmedetomidine for labor analgesia: A prospective double-blinded study.

BACKGROUND: Studies have shown that the ideal dose of epidural dexmedetomidine is 0.4 µg mL-1 for epidural labor analgesia. However, the appropriate dose of ropivacaine when combined with 0.4 µg mL-1 of dexmedetomidine for epidural labor analgesia is still unknown. Therefore, we aimed to determine the dose-response of ropivacaine when using 0.4 µg mL-1 of dexmedetomidine as epidural adjuvant for labor analgesia. METHODS: One hundred of nulliparous singleton pregnant patients were randomized allocated into 1 of 5 groups with epidural ropivacaine concentration of 0.05%, 0.0625%, 0.075%, 0.0875%, and 0.1%. Labor analgesia was initialed with 12 mL of the mixed study solution. Effective analgesia was defined as a visual analogue scale <10 mm 30 minutes after the initial epidural bolus. The EC50 and EC95 for epidural ropivacaine was calculated by probit regression. RESULTS: Ninety-three of parturients were involved into the final analysis. Totals of 63.2% (12/19), 73.7% (14/19), 88.9% (16/18), 94.7% (18/19), and 100% (18/18) of parturients in group 0.05, 0.0625, 0.075, 0.0875, and 0.1 received effective epidural labor analgesia. The calculated EC50 and EC95 of epidural ropivacaine were 0.046% (95% CI 0.028-0.054%) and 0.086% (95% CI 0.074-0.137%), respectively. CONCLUSIONS: Under the condition of the study, a bolus of 12 mL ropivacaine 0.086% and dexmedetomidine 0.4 µg mL-1 could afford 95% of nulliparous singleton pregnant patients without suffering labor pain after a test dose of lidocaine 45 mg.

Removal of Fermentation Inhibitors from Sugarcane Bagasse Hydrolysate via Post-cross-linked Hydrophilic-Hydrophobic Interpenetrating Polymer Networks.

The efficient and economical removal of fermentation inhibitors from the complex system of biomass hydrolysate was one of the basics and keys in bio-chemical transformation. In this work, post-cross-linked hydrophilic-hydrophobic interpenetrating polymer networks (PMA/PS_pc IPNs and PAM/PS_pc IPNs) were proposed to remove fermentation inhibitors from sugarcane bagasse hydrolysate for the first time. PMA/PS_pc and PAM/PS_pc IPNs can obviously enhance the adsorption performance towards fermentation inhibitors due to their higher surface area and hydrophilic-hydrophobic synergetic surface properties, especially PMA/PS_pc IPNs has higher selectivity coefficients of 4.57, 4.63, 4.85, 16.0, 49.43, and 22.69, and higher adsorption capacity of 24.7 mg/g, 39.2 mg/g, 52.4 mg/g, 9.1 mg/g, 13.2 mg/g, and 144.9 mg/g towards formic acid, acetic acid, levulinic acid (LA), 5-hydroxymethylfurfural (HMF), furfural, and acid-soluble lignin (ASL), respectively, in a lower total sugar loss of 2.03%. The adsorption kinetics and isotherm of PMA/PS_pc IPNs were studied to elucidate its adsorption behavior towards fermentation inhibitors. In addition, the cyclic utilization property of PMA/PS_pc IPNs was stable. Synthesizing PMA/PS_pc IPNs is a new strategy to provide an efficient adsorbent for the removal of fermentation inhibitors from lignocellulosic hydrolysate.

Immobilization of Acetobacter sp. CCTCC M209061 for efficient asymmetric reduction of ketones and biocatalyst recycling.

BACKGROUND: The bacterium Acetobacter sp. CCTCC M209061 is a promising whole-cell biocatalyst with exclusive anti-Prelog stereoselectivity for the reduction of prochiral ketones that can be used to make valuable chiral alcohols such as (R)-4-(trimethylsilyl)-3-butyn-2-ol. Although it has promising catalytic properties, its stability and reusability are relatively poor compared to other biocatalysts. Hence, we explored various materials for immobilizing the active cells, in order to improve the operational stability of biocatalyst. RESULTS: It was found that Ca-alginate give the best immobilized biocatalyst, which was then coated with chitosan to further improve its mechanical strength and swelling-resistance properties. Conditions were optimized for formation of reusable immobilized beads which can be used for repeated batch asymmetric reduction of 4'-chloroacetophenone. The optimized immobilized biocatalyst was very promising, with a specific activity of 85% that of the free-cell biocatalyst (34.66 µmol/min/g dw of cells for immobilized catalyst vs 40.54 µmol/min/g for free cells in the asymmetric reduction of 4'-chloroacetophenone). The immobilized cells showed better thermal stability, pH stability, solvent tolerance and storability compared with free cells. After 25 cycles reaction, the immobilized beads still retained >50% catalytic activity, which was 3.5 times higher than degree of retention of activity by free cells reused in a similar way. The cells could be recultured in the beads to regain full activity and perform a further 25 cycles of the reduction reaction. The external mass transfer resistances were negligible as deduced from Damkohler modulus Da < <1, and internal mass transfer restriction affected the reduction action but was not the principal rate-controlling step according to effectiveness factors η < 1 and Thiele modulus 0.3<∅ <1. CONCLUSIONS: Ca-alginate coated with chitosan is a highly effective material for immobilization of Acetobacter sp. CCTCC M209061 cells for repeated use in the asymmetric reduction of ketones. Only a small cost in terms of the slightly lower catalytic activity compared to free cells could give highly practicable immobilized biocatalyst.

Microbial oil production from corncob acid hydrolysate by Trichosporon cutaneum.

Corncob was treated by dilute H(2)SO(4). The hydrolysate contained 45.7 g sugar/l. Without concentration or adding other nutrients, the hydrolysate, after being detoxified by overliming and adsorption with activated charcoal, was used for oil production using Trichosporon cutaneum. After 8 days' growth in shake-flasks, the biomass was 22.1 g/l with a lipid content of 36%. The lipid yield per mass of sugar was 17.4% (w/w). Corncob thus is a promising raw material for microbial oil production by this yeast.

Attapulgite Structure Reset to Accelerate the Crystal Transformation of Isotactic Polybutene.

Isotactic polybutene (iPB) has a wide application in the water pipe field. However, the most valuable form I, needs 7 days to complete the transformation. In this study, the attapulgite (ATP), which produces lattice matching of the iPB form I, was selected to prepare an iPB/ATP composite. The Fischer-Tropsch wax (FTW) was grafted with maleic anhydride to obtain MAFT, and the ATP structure was reset by reactions with MAFT to the prepared FATP, which improved the interface compatibility of the ATP and iPB. The Fourier transform infrared spectroscopy (FT-IR) and the water contact angle test confirmed the successful synthesis of FATP. X-ray diffraction (XRD) verified that the graft of MAFT did not affect the crystal structure of ATP. The iPB + 5% FATP had the maximum flexural strength, which was 12.45 Mpa, and the flexural strength of the iPB + 5% FATP annealing for 1 day was much higher than others. Scanning electron microscope (SEM) photographs verified that FATP and iPB had good interface compatibility. The crystal transformation behavior indicated that the iPB + 5% FATP had the fastest crystal transformation rate, which proved that the reset structure, ATP, greatly accelerated the crystal transformation of iPB. This was a detailed study on the effect of lattice matching, interfacial compatibility and internal lubrication of the reset structure, ATP, in the nucleation and growth stages of iPB form I. The result was verified by XRD, differential scanning calorimetry (DSC), Avrami kinetics and polarizing microscope (POM) analysis.

Synthesis of polyacrylamide/polystyrene interpenetrating polymer networks and the effect of textural properties on adsorption performance of fermentation inhibitors from sugarcane bagasse hydrolysate.

Economical removal of fermentation inhibitors from lignocellulosic hydrolysate plays a considerable role in bioconversion of lignocellulose biomass. In this work, the textural properties of polyacrylamide/polystyrene interpenetrating polymer networks (PAM/PS IPNs) on adsorption of fermentation inhibitors from sugarcane bagasse hydrolysate (SCBH) were investigated for the first time. The results showed that, the specific surface area, pore diameter and surface polarity had important influence on its adsorption performance towards sugars, organic acids, furans and acid-soluble lignin. The PAM/PS IPNs under the optimal copolymerization situation achieved the high selectivity coefficients of 4.07, 14.9, 21.2 and 25.8 with respective to levulinic acid, furfural, hydroxymethylfurfural (HMF) and acid-soluble lignin, and had a low total sugar loss of 2.09%. Overall, this research puts forward a design and synthetic strategy for adsorbent to remove fermentation inhibitors from lignocellulosic hydrolysate.

Anaerobic digestion of elephant grass hydrolysate: Biogas production, substrate metabolism and outlet effluent treatment.

Elephant grass (Pennisetum purpureum) acid hydrolysate was used as substrate for anaerobic digestion for the first time. Within short period (ten days), the organic materials (sugars and organic acids) in the elephant grass hydrolysate could be utilized efficiently for stable biogas production that the COD removal, biogas yield, and CH4 content were 91.3 ±â€¯2.0%, 0.561 ±â€¯0.014 m3/kg COD consumption, and 68.1 ±â€¯4.6%, respectively throughout this bioprocess. During anaerobic digestion, almost no volatile fatty acids (VFAs) was accumulated (merely <0.1 g/L acetic acid was found) and the outlet pH was very stable (7.3 ±â€¯0.1). Meanwhile, the furans including furfural and 5-hydroxymethylfurfural (HMF) existing in the inlet substrate could be degraded. After anaerobic digestion, the outlet effluent was treated by combination of Fe-C micro-electrolysis and Fenton reaction to remove 93.1% of residual COD and 98.6% of color. Considering the performance, cost, operation, and environmental influence, this technology is suitable for industrial treatment of waste elephant grass.

Introduction of one efficient industrial system for turpentine processing wastewater reuse and treatment.

Wastewater treatment is one important issue for turpentine plant and more wastewater generated by greater turpentine processing will prevent its further development. To solve this issue without extra place and new equipment, one industrial system for reuse and treatment of turpentine processing wastewater was introduced for the first time. For wastewater reuse, the technology was simple and easy to control that after neutralization by lime and absorption with activated carbon (optional, mostly not necessary), the wastewater could be reused for turpentine processing. After reuse, the wastewater was further treated by a biological system. During long-term application of wastewater reuse in this plant, it showed little influence on the products performance (mainly acid value) and final wastewater COD. Base on above advantages, the plant could decide when for wastewater drainage, and thus the amount of wastewater was reduced greatly. For the biological treatment, the COD of wastewater could be degraded to suitable level stably and the wastewater after treatment could be applied for daily life in the plant. Overall, considering the cost, operation, and performance, the whole system shows great potential and possibility of industrial application and therefore can be applied widely in the turpentine processing industry.

Bacterial cellulose based superabsorbent production: A promising example for high value-added utilization of clay and biology resources.

Superabsorbent was synthesized from bacterial cellulose (BC) generated by in situ fermentation on bentonite inorganic gel (BIG). For BIG preparation, the effect of sodium agent's type and content, temperature and time of sodium-modification, and gelling agent's type and content on the viscosity of BIG were learned to optimize the synthesis process. For polymerization, the effect of different factors including ratio of monomer to substrate (modified BC from in situ fermentation), content of initiator and crosslinker, monomer neutralization degree, reaction temperature and time on the performance of composite (superabsorbent) synthesized were analyzed. Under optimal condition, the composite showed good water absorption, salts absorption, and water retention capacity. The original bentonite, sodium-based bentonite, BIG and composite structure was characterized by X-ray fluorescence (XRF), nuclear magnetic resonance (NMR), Fourier transform infrared spectroscopy (FT-IR), scanning electron microscope (SEM) and thermogravimetric analysis (TGA), and the characterization partly explained the performance of water absorption and thermal stability of the composite. Overall, this study provides one method for superabsorbent synthesis from low-cost and natural resources.

Adsorption Study of Acid Soluble Lignin Removal from Sugarcane Bagasse Hydrolysate by a Self-Synthesized Resin for Lipid Production.

An adsorption resin CX-6 was synthesized and used for acid soluble lignin (ASL) removal from sugarcane bagasse hydrolysate (SCBH). The adsorption conditions of pH value, amount of adsorbent, initial ASL concentration, and temperature on ASL adsorption were discussed. The results showed the adsorption capacity of ASL was negatively affected by increasing temperature, solution pH, and adsorbent dose, and was positively affected by increasing initial concentration. The maximum adsorption capacity of ASL was 135.3 mg/g at initial ASL concentration 6.46 g/L, adsorption temperature 298 K, and pH 1. Thermodynamic study demonstrated that the adsorption process was spontaneous and exothermic. Equilibrium and kinetics experiments were proved to fit the Freundlich isotherm model and pseudo-second-order model well, respectively. Fermentation experiment showed that the SCBH after combined overliming with resin adsorption as fermentation substrate for microbial lipid production by Trichosporon cutaneum and Trichosporon coremiiforme was as better as that of SCBH by combined overliming with active charcoal adsorption, and more efficient than that of SCBH only by overliming. Moreover, the regeneration experiment indicated that the CX-6 resin is easy to regenerate and its recirculated performance is stable. In conclusion, our results provide a promising adsorbent to detoxify lignocellulose hydrolysate for further fermentation.