We provide further insight into the photochemical control of viscoelasticity through the use of azobenzene sodium dicarboxylate for molecular switching. As a photoresponsive molecule, Sodium 3,3'-azobenzene dicarboxylate (3,3'-Azo2Na) was added to a solution of cetyltrimethylammonium bromide (CTAB)/sodium salicylate (NaSal), which is known for inducing the formation of wormlike micelles. This solution maintained a wormlike micellar structure, although a reduction in zero-shear viscosity was observed. When this mixed aqueous solution of CTAB/NaSal/3,3'-Azo2Na (16.7 mM each) was irradiated by ultraviolet light, the 3,3'-Azo2Na exhibited molecular trans-cis photoisomerization. We measured the dynamic viscoelasticity of the sample in the photostationary state and found that the zero-shear viscosity increased approximately sevenfold compared to the preirradiation state. This phenomenon is the opposite of the system wherein viscosity reduced by irradiation, as reported by us. We discuss the mechanism of this viscosity change.
Although liposomes are considered to be one of the most promising carriers for drug delivery systems (DDS), they have drawbacks such as insufficient drug-entrapment efficiency and long-term stability. The objectives of this study are to improve the trapping efficiency by addition of lipopeptides (LPs), and using a supercritical CO2 reverse-phase evaporation (SCRPE) process, along with incorporation of PEG-modified phospholipids to improve long-term stability. In this study, bovine serum albumin (BSA) was used as a model drug substance for entrapment by liposomes. Improvements in the entrapment efficiency and stability of liposomes were achieved by modification with LPs and use of a SCRPE preparation process. The BSA-entrapment efficiency of liposomes modified with cationic LPs with arginine residues, as a result of their ionic interactions, was six times that of liposomes prepared by the Bangham method. Use of a SCRPE method along with LP modification further enhanced entrapment and enabled spontaneous formation of unilamellar liposomes with long-term stability. Liposomes consisting of DPPC/Chol/C16-Arg2/DSPE-PEG2000 (60/30/5/5), with up to 70% entrapment efficiency for BSA and a stability level of 90% for over 40 h, were obtained. DSC and SAXS analyses indicated that certain amounts of LP in the DPPC induced phase-transitional and structural changes in the lamellar membrane, and these changes improved the DDS carrier properties. The SCRPE method provides organic-solvent-free liposomes, and the LPs for the liposome modification are derivatives of amino acids and fatty acids, which are sustainable and biocompatible materials. This study therefore suggests that there are opportunities for the development of novel DDS carriers with excellent performance and which address environmental concerns.
To examine the effect of cold shock treatment on the fatty acid composition of Aurantiochytrium limacinum strain mh0186, a marine thraustochytrid, we cultivated this strain at 28°C for 72 h with shaking and stored the obtained biomass at 10°C for 72 h. A growth experiment was carried out for comparison, wherein strain mh0186 was grown at 10 and 15°C for 72 h with shaking, and it was found that the unsaturation of fatty acids was accelerated relative to that at 28°C. In the cold shock experiment, the total lipid content significantly increased during storage at 10°C for 72 h. Overall, the percentage of unsaturated fatty acids such as docosahexaenoic acid was almost stable while that of n-6 docosapentaenoic acid decreased slightly, but significantly, relative to that in the growth experiment.
Enoyl-coenzyme A (CoA) hydratase catalyzes the hydration of trans-2-enoyl-CoA to yield 3-hydroxyacyl-CoA during fatty acid degradation (β-oxidation). Although much research has focused on the stereospecificities of 2-enoyl-CoA hydratases, a direct quantification of the production of 3(R)- and 3(S)-hydroxyacyl-CoA has not yet been established. Therefore, we developed a method of concurrently quantifying 3(R)- and 3(S)-hydroxyacyl-CoA using high-performance liquid chromatography (HPLC) equipped with a chiral separation column. The optimized conditions for the separation of 3(R)-, 3(S)-hydroxyhexadecanoyl-CoA and trans-2-hexadecenoyl-CoA, were determined to be as follows: mobile phase of 35/65 (v/v) of 50 mM phosphate buffer (pH 5.0)/methanol; flow rate of 0.5 mL/min; detection at 260 nm; and column temperature of 25°C. This method was applied to subcellular fractions of rat liver; the results directly confirmed that 3(S)-hydroxyhexadecanoyl-CoA is the dominant product obtained from the heat-stable enoyl-CoA hydratase-catalyzed reaction of trans-2-hexadecenoyl-CoA. Finally, the stereospecificities of L-bifunctional protein (L-BP) and D-bifunctional protein (D-BP) were reinvestigated using this method, and it was confirmed that L- and D-BP yielded 3(S)- and 3(R)-hydroxyhexadecanoyl-CoA were yielded from trans-2-hexadecenoyl-CoA, respectively. 3(R)-Hydroxyacyl-CoA is a peroxisomal β-oxidation-specific intermediate. Therefore, this method is potentially useful not only studies regarding the stereochemistry of enoyl-CoA hydratase but also for the diagnosis of diseases caused by defects of peroxisomal enoyl-CoA hydratase.
The secondary structure of bovine serum albumin (BSA) in the binary surfactant system of anionic sodium dodecyl sulfate (SDS) and zwitterionic N-dodecyl-N,N-dimethyl-3-ammonio-1- propanesulfonate (DDAPS) was examined at 25°C. The helicity of BSA decreased from 66% to 55% in a solution of DDAPS alone and decreased to 50% in a solution of SDS alone. However, the late addition of DDAPS reformed the helical structure of BSA, which was initially disrupted by SDS. The reformation required higher DDAPS concentrations as the initial SDS concentration increased. A maximum helicity of 63% was attained by this reformation. On the other hand, the helical structure of the protein, which was first affected by DDAPS denaturation, was also reformed to the same degree by the late addition of certain amounts of SDS. Although attention was paid to the additive order of these two surfactants to BSA, the final helicity of the protein depended on the final concentrations of these two surfactants, irrespective of the additive order. These phenomena may be attributed to the predominance of mixed micelle formation over complex formation between BSA and the two surfactants below the mixing ratio of DDAPS ([DDAPS]/([DDAPS]+[SDS])) of 0.95. The predominance of the mixed micelle formation distinctly appeared in mixing ratios between 0.50 and 0.75. In this range, the mixed micelle formation accompanied the removal of dodecyl sulfate (DS) ions bound to BSA upon the late addition of DDAPS to the BSA-SDS mixture, whereas, upon the late addition of SDS to the BSA-DDAPS mixture, the mixed micelle formation was accelerated by the coexistence of DDAPS which disturbed the binding of DS ions to the protein.
Hydroxyl fatty acids and their derivatives are of high value due to their wide range of industrial application, including cosmetic, food, personal care and pharmaceutical products. Realizing the importance of hydroxyl fatty acids, and yet due to the absence of the conventional starting raw materials, Malaysia has developed 9,10-dihydroxystearic acid (9,10-DHSA) and its derivatives from locally abundant palm based oils. The aim of this article is to provide a general description of the works that have thus far being done on palm based 9,10-DHSA: starting from its conception and production from commercial grade palm based crude oleic acid via epoxidation and hydrolysis, purification through solvent crystallization and characterization through wet and analytical chemistry, moving on to developmental works done on producing its derivatives through blending, esterification, amidation and polymerization, and completing with applications of 9,10-DHSA and its derivatives, e.g. DHSA-stearates and DHSA-estolides, in commercial products such as soaps, deodorant sticks and shampoos. This article incorporates some of the patent filed technological knowhow on 9,10-DHSA and its derivatives, and will also point out some of the shortcomings in previously published documents and provide some recommendations for future research works in mitigating these shortcomings.
Biosurfactants (BS) are produced by a variety of microorganisms from renewable resources, and have unique properties compared to chemical surfactants. In order to attain efficient production of BS from low-cost materials, we focused our attention on the use of sugarcane molasses. Fifteen yeast strains that are known as BS producers were examined for BS productivity from a culture medium consisting of only molasses and water. Among the strains tested, only Starmerella bombicola NBRC 10243 produced sophorolipids (SL), which are glycolipid BS. The culture conditions for the yeast were then investigated in a shake-flask culture. SL production was significantly affected by the pH of the medium and was highly accelerated at pH 6. Under the optimum conditions, the amount of SL reached 14.4 g/L after 120 h from a medium containing 150 g/L of total sugars. We tried to improve the production of SL further by feeding the molasses using a jar fermentor. Interestingly, the amount of SL increased up to 22.8 g/L after 120 h; the production rate was 1.6-fold higher than that in the shake-flask culture. These results suggest that the present yeast should have great potential for the low-cost production of SL, and facilitate the application of BS in various fields.