Characterization of acetylated starch nanoparticles for potential use as an emulsion stabilizer.

To overcome the low production efficiency of Pickering emulsion stabilizers prepared from starch, alcohol precipitation and surface modification were applied in this study. Spherical starch nanoparticles (StNPs) (247.90 ± 1.96 nm) were prepared through nanoprecipitation. The StNPs were surface-esterified to produce starch nanoparticle acetate (StNPAc), and the physicochemical changes of the products were investigated. The contact angle (>89.56° ± 0.56°) of StNPAc (degree of substitution, 0.53) was maintained for over 30 min. The results showed that the hydrophobicity of the StNPs was improved by shielding the surface hydroxyl groups via acetylation. StNPAc was also used to produce emulsions for further evaluation of their feasibility as Pickering emulsifiers. Oil-in-water (3:7, v/v) emulsions containing 1.5 wt% StNPAc were stabilized for over 35 days without creaming. Thus, StNPAc exhibited better emulsifying capacity and storage stability than StNPs. Therefore, hydrophobic starch nanoparticles obtained by acetylation are promising stabilizers for surfactant-free Pickering emulsions.

Optimization of monomethoxy polyethyleneglycol-modified oxalate decarboxylase by response surface methodology.

In order to improve the stability of oxalate decarboxylase (Oxdc), response surface methodology (RSM), based on a four-factor three-level Box-Behnken central composite design was used to optimize the reaction conditions of oxalate decarboxylase (Oxdc) modified with monomethoxy polyethyleneglycol (mPEG5000). Four independent variables such as the ratio of mPEG-aldehyde to Oxdc, reaction time, temperature, and reaction pH were investigated in this work. The structure of modified Oxdc was identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Fourier transform infrared (FTIR) spectroscopy, the stability of the modified Oxdc was also investigated. The optimal conditions were as follows: the mole ratio of mPEG-aldehyde to Oxdc of 1:47.6, time of 13.1 h, temperature at 29.9 °C, and the reaction pH of 5.3. Under optimal conditions, experimental modified rate (MR = 73.69%) and recovery rate (RR = 67.58%) were matched well with the predicted value (MR = 75.11%) and (RR = 69.17%). SDS-PAGE and FTIR analysis showed that mPEG was covalently bound to the Oxdc. Compared with native Oxdc, the modified Oxdc (mPEG-Oxdc) showed higher thermal stability and better tolerance to trypsin or different pH treatment. This work will provide a further theoretical reference for enzyme modification and conditional optimization.

Chemical modification of oxalate decarboxylase to improve adsorption capacity.

In order to enhance the adsorption capacity of oxalate decarboxylase (Oxdc) on calcium oxalate monohydrate crystals and improve the application performance of Oxdc, chemical modification of Oxdc with ethylenediaminetetraacetic dianhydride (EDTAD) was investigated in this work. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and liquid chromatography tandem mass spectrometry (LC/MS) analysis results demonstrated that Oxdc and EDTAD have been covalently bound, and suggested that the chemical modification occurred at the free amino of the side chain and the α-amine of the N-terminus of Oxdc. Fluorescene and circular dichroic measurement showed that the structure and conformation of Oxdc were tinily altered after modification by EDTAD. The optimum pH of EDTAD-modified Oxdc was shifted to the alkaline side about 1.5 unit and it has a higher thermostability. The analysis of kinetic parameters indicated that the EDTAD-modified Oxdc showed a higher affinity towards the substrate. Through modification the adsorption capacity of Oxdc onto CaOx monohydrate crystals was increased by 42.42%.

Adsorption characteristics of amino acids on to calcium oxalate.

Adsorption of amino acids on to calcium oxalate found in urinary calculus has been studied and the adsorption characteristics were analyzed. Pseudo-first-order, pseudo-second-order and intraparticle diffusion models were used to fit the kinetics data. The pseudo-second-order model best described the dynamic behavior of the adsorption process. The uptake of glutamic acid and aspartic acid were found to decrease as solution pH increasing from 4 to 8. The experimental data obtained at different pH conditions were analyzed and fitted by Langmuir, Freundlich, Redlich-Peterson, Temkin and Sips isotherm models using linear and nonlinear regression analysis. Error analysis (correlation coefficient, residual root mean square error and chi-square test) showed that the Langmuir I isotherm model and the non-linear form of Sips isotherm model should be primarily adopted for fitting the equilibrium data. The maximum adsorption capacity of glutamic acid and aspartic acid onto calcium oxalate monohydrate crystals are 0.059 and 0.066µmol/g at pH 4, respectively. These studies have the vital significance for research aimed at exploring the role of urinary amino acids effect the formation process of calcium oxalate crystals found in urinary calculus and for potential application in the design of synthetic peptides used for urinary calculi therapy.

Synthesis of rosin acid starch catalyzed by lipase.

Rosin, an abundant raw material from pine trees, was used as a starting material directly for the synthesis of rosin acid starch. The esterification reaction was catalyzed by lipase (Novozym 435) under mild conditions. Based on single factor experimentation, the optimal esterification conditions were obtained as follows: rosin acid/anhydrous glucose unit in the molar ratio 2:1, reaction time 4 h at 45°C, and 15% of lipase dosage. The degree of substitution (DS) reaches 0.098. Product from esterification of cassava starch with rosin acid was confirmed by FTIR spectroscopy and iodine coloration analysis. Scanning electron microscopy and X-ray diffraction analysis showed that the morphology and crystallinity of the cassava starch were largely destroyed. Thermogravimetric analysis indicated that thermal stability of rosin acid starch decreased compared with native starch.

Immobilization of oxalate decarboxylase to Eupergit and properties of the immobilized enzyme.

Oxalate decarboxylase, an oxalate degradation enzyme used for medical diagnosis and decreasing the oxalate level in the food or paper industry, was covalently immobilized to Eupergit C. Different immobilization parameters, including ratio of enzyme to support, ammonia sulfate concentration, pH, and incubation time, were optimized. Under the condition of enzyme/support ratio at 1:20, pH 9, with 1.5 mol/L (NH(4))(2)SO(4), room temperature, and shaking at 30 rpm for 24 hr, activity recovery of immobilized Oxdc reached 90% with an apparent specific activity of 0.44 U/mg support. The enzymatic properties of immobilized Oxdc were investigated and compared with those of the soluble enzyme. Both shared a similar profile of optimum conditions; the optimum pH and temperature for soluble and immobilized Oxdc were 3.5 and 50°C, respectively. The immobilized enzyme was more stable at lower pH and higher temperatures. The kinetic parameters for soluble and immobilized enzyme were also determined.

Case report: intraoperative management of extreme hemodilution in a patient with a severed axillary artery.

We present a case of extreme hemodilution in which appropriately crossmatched blood was not available. A 53-year-old man was admitted to our hospital because of hemorrhagic shock due to multiple stab wounds. His blood type was B, Rh negative, and his intravascular fluid volume was maintained with balanced salt solution and plasma substitutes, i.e., hydroxyethyl starch. His hemoglobin reached a nadir of 0.7 g/dL and hematocrit 2.2% before being transfused. No evidence of cardiac ischemia was noted and he was discharged in good condition. Extreme hemodilution can be successfully managed by maintaining a normal blood volume, 100% oxygen, and the use of plasma substitutes.

3-Hydroxypropionaldehyde guided glycerol feeding strategy in aerobic 1,3-propanediol production by Klebsiella pneumoniae.

3-Hydroxypropionaldehyde (3-HPA) is a toxic intermediary metabolite in the biological route of 1,3-propanediol biosynthesis from glycerol. 3-HPA accumulated in culture medium would arouse an irreversible cessation of the fermentation process. The role of substrate (glycerol) on 3-HPA accumulation in aerobic fermentation was investigated in this paper. 1,3-Propanediol oxidoreductase and glycerol dehydratase, two key enzyme catalyzing reactions of 3-HPA formation and consumption, were sensitive to high concentration of 3-HPA. When the concentration of 3-HPA increased to a higher level in medium (ac 10 mmol/L), the activity of 1,3-propanediol oxidoreductase in cell decreased correspondingly, which led to decrease of the 3-HPA conversion rate, then the 3-HPA concentration increasing was accelerated furthermore. 3-HPA accumulation in culture medium was triggered by this positive feedback mechanism. In the cell exponential growth phase, the reaction catalyzed by 1,3-propanediol oxidoreductase was the rate limiting step in 1,3-propanediol production. The level of 3-HPA in culture medium could be controlled by the substrate (glycerol) concentration, and lower level of glycerol could avoid 3-HPA accumulating to a high, lethal concentration. In fed batch fermentation, under the condition of initial glycerol concentration 30 g/L, and keeping glycerol concentration lower than 7-8 g/L in cell exponential growth phase, 3-HPA accumulation could not be incurred. Based on this result, a glycerol feeding strategy was set up in fed batch fermentation. Under the optimized condition, 50.1 g/L of 1,3-propanediol was produced in 24 h, and 73.1 g/L of final 1,3-propanediol concentration was obtained in 54 h.

Enhancement of 1,3-propanediol production by Klebsiella pneumoniae with fumarate addition.

Addition of 5 mM: fumarate to cultures of Klebsiella pneumoniae enhanced the rate of glycerol consumption and the production of 1,3-propanediol (PDO). Compared to the control, the activity of glycerol dehydrogenase increased by 35, 33 and 46%, the activity of glycerol dehydratase increased by 160, 210 and 115%, and the activity of 1,3-propanediol oxidoreductase increased by 25, 39 and 85% when, respectively, 5, 15 and 25 mM: fumarate were provided. At the same time, the ratio of NAD+ to NADH decreased by 20, 23 and 29%. Using a 5 l bioreactor with 5 mM: fumarate addition, the specific rate of glycerol consumption and the productivity of PDO was 30 mmol/l h and 17 mmol/l h, respectively, both increased by 35% over the control.