Facile purification of tocopherols from soybean oil deodorizer distillate in high yield using lipase

Tocopherols have been purified from deodorizer distillate produced in the final deodorization step of vegetable oil refining by a process including molecular distillation. Deodorizer distillate contains mainly tocopherols, sterols, and free fatty acids (FFA); the presence of sterols hinders tocopherol purification in good yield. We found that Candida rugosa lipase recognized sterols as substrates but not tocopherols, and that esterification of sterols with FFA could be effected with negligible influence of water content. Enzymatic esterification of sterols with FFA was thus used as a step in tocopherol purification. High boiling point substances including steryl esters were removed from soybean oil deodorizer distillate by distillation, and the resulting distillate (soybean oil deodorizer distillate tocopherol concentrate; SODDTC) was used as a starting material for tocopherol purification. Several factors affecting esterification of sterols were investigated, and the reaction conditions were determined as follows: A mixture of SODDTC and water (4∶1, w/w) was stirred at 35°C for 24 h with 200 U of Candida lipase per 1 g of the reaction mixture. Under these conditions, approximately 80% of sterols was esterified, but tocopherols were not esterified. After the reaction, tocopherols and FFA were recovered as a distillate by molecular distillation of the oil layer. To enhance further removal of the remaining sterols, the lipase-catalyzed reaction was repeated on the distillate under the same reaction conditions. As a result, more than 95% of the sterols was esterified in total. The resulting reaction mixture was fractionated to four distillates and one residue. The main distillate fraction contained 65 wt% tocopherols with low contents of FFA and sterols. In addition, the residue fraction contained high-purity steryl esters. Because the process presented in this study includes only organic solvent-free enzymatic reaction and molecular distillation, it is feasible as a new industrial purification method of tocopherols. This work was presented at the Biocatalysis symposium in April 2000, held at the 91st Annual Meeting and Expo of the American Oil Chemists Society, San Diego, CA.     

Ghosts of B‐type Wheat Starch Granules in Concentrated KI/I2 Solution

When B‐type (2.0–8.0 µm) wheat starch granules containing various amounts (2.1–25%) of amylose were treated with 25% KI/10% I₂ solution, low‐amylose (below 10% amylose) B‐type wheat starch granules changed to the ghost form. It is known that A‐type (25–35 µm) wheat starch granules change to the ghost form and show a typical double structure (a black‐brown central portion and red‐brown surrounding portion), however, the B‐type wheat starch ghosts did not show the same double structure but rather a simple (red‐brown portion) sack form. The relative ghost areas in the B‐type wheat starch granules were highly correlated to the amylose content (%), which was similar to the results of A‐type starch granules. This suggests that the amylose molecule in B‐type starch contributes to the structural stability of the starch granule.     

Time-resolved DXAFS study on the reduction processes of Cu cations in ZSM-5

The time-resolved reduction process of copper cations in/on ZSM-5 during temperature-programmed reduction (300–700 K) was studied by energy-dispersive X-ray absorption fine structure (DXAFS) as well as transmission electron microscopy (TEM). Two Cu-ZSM-5 samples with different Cu loadings were prepared by an ion-exchange method. The Cu K-edge DXAFS spectra for isolated Cu₂₊ species in the channels of ZSM-5 and CuO particles on the outer surfaces of ZSM-5 were recorded at an interval of 1 s during the reduction. The curve fitting analysis of the EXAFS data and the XANES analysis revealed that the isolated Cu₂₊ species in the channels were reduced stepwise. They were reduced to isolated Cu₊ species at 400–450 K and the Cu₊ species to Cu₀ metallic clusters at 550–650 K. Small clusters like Cu₄ were initially formed, followed by particle growth. A small part of them went out to the outer surfaces of ZSM-5 during the reduction. In contrast, CuO particles on the outer surfaces were reduced directly to Cu₀ metallic particles around 450 K.     

Extension of the applicable range of the implicit curve-fitting method for refrigerant thermodynamic properties to critical pressure

The method of implicit curve-fitting and explicit-calculation has been used for fast and stable calculations of thermodynamic properties of subcritical refrigerants. In order to extend that method to the critical pressure, a method of sectional implicit curve-fitting and explicit-calculation for refrigerant thermodynamic properties is introduced in this paper. The whole data range is divided into several subsections. The requirements on the continuity of thermodynamic properties and the first order derivative of thermodynamic properties in the intersection points of subsections are indicated, and the methods to meet the requirements are presented. Quadric equations are constructed instead of curve-fitting when no data can be given. With the source data obtained from REFPROP 7.1, explicit fast calculation formulae for thermodynamic properties of R410A, covering the saturated temperature of 213.15–344.51 K and superheat of 0–65 K, are given as an example. The calculation speeds of the formulae of R410A are more than 7000 times faster than those of REFPROP 7.1 while the total mean relative deviation of the fast calculation formulae from REFPROP 7.1 is only 0.04%.     

Elevated Expression of CCN3 in Articular Cartilage Induces Osteoarthritis in Hip Joints Irrespective of Age and Weight Bearing

Osteoarthritis (OA) occurs not only in the knee but also in peripheral joints throughout the whole body. Previously, we have shown that the expression of cellular communication network factor 3 (CCN3), a matricellular protein, increases with age in knee articular cartilage, and the misexpression of CCN3 in cartilage induces senescence-associated secretory phenotype (SASP) factors, indicating that CCN3 promotes cartilage senescence. Here, we investigated the correlation between CCN3 expression and OA degenerative changes, principally in human femoral head cartilage. Human femoral heads obtained from patients who received total hip arthroplasty were categorized into OA and femoral neck fracture (normal) groups without significant age differences. Gene expression analysis of RNA obtained from femoral head cartilage revealed that CCN3 and MMP-13 expression in the non-weight-bearing part was significantly higher in the OA group than in the normal group, whereas the weight-bearing OA parts and normal cartilage showed no significant differences in the expression of these genes. The expression of COL10A1, however, was significantly higher in weight-bearing OA parts compared with normal weight-bearing parts, and was also higher in weight-bearing parts compared with non-weight-bearing parts in the OA group. In contrast, OA primary chondrocytes from weight-bearing parts showed higher expression of CCN3, p16, ADAMTS4, and IL-1β than chondrocytes from the corresponding normal group, and higher ADAMTS4 and IL-1β in the non-weight-bearing part compared with the corresponding normal group. Acan expression was significantly lower in the non-weight-bearing group in OA primary chondrocytes than in the corresponding normal chondrocytes. The expression level of CCN3 did not show significant differences between the weight-bearing part and non-weight-bearing part in both OA and normal primary chondrocytes. Immunohistochemical analysis showed accumulated CCN3 and aggrecan neoepitope staining in both the weight-bearing part and non-weight-bearing part in the OA group compared with the normal group. The CCN3 expression level in cartilage had a positive correlation with the Mankin score. X-ray analysis of cartilage-specific CCN3 overexpression mice (Tg) revealed deformation of the femoral and humeral head in the early stage, and immunohistochemical analysis showed accumulated aggrecan neoepitope staining as well as CCN3 staining and the roughening of the joint surface in Tg femoral and humeral heads. Primary chondrocytes from the Tg femoral head showed enhanced expression of Ccn3, Adamts5, p16, Il-6, and Tnfα, and decreased expression of Col2a1 and -an. These findings indicate a correlation between OA degenerative changes and the expression of CCN3, irrespective of age and mechanical loading. Furthermore, the Mankin score indicates that the expression level of Ccn3 correlates with the progression of OA.     

The associative nature of lower rank coal

Initial volumetric swelling in tetrahydrofuran of pyridine-unextracted parts from subbituminous coal and lignite showed no dependence of their concentration, and was smaller than that of their pyridine extracts. These results are opposite to those obtained from high volatile bituminous coals and coincide with predictions for the cross-linked network model of coal. However, when ionic forces in these coals were reduced by acid washing or O-alkylation, these coals showed the same associative nature as did high volatile bituminous coals. Swelling kinetics were analysed on the basis of associative equilibria controlled by the ionic forces. It was concluded that solvation of the ionic forces was the rate determining step of volumetric swelling of lower rank coal rather than solvent diffusion into the coal, although diffusion has been proposed to be the most important factor in swelling.     

Fatty acid-containing lipids of the yeast Saccharomyces cerevisiae during post-fermentation decline in viability

On exhaustion of the sugar supply in the medium, cells of the yeast Saccharomyces cerevisiae maintained at 30°C lost most of their phospholipid content and their viability, as assessed by methylene blue staining, within a few days. Both processes occurred more rapidly in cells which had fermented glucose rather than maltose. During this period of decline the cell content of other fatty acid esters, mainly t riacylglycerols, increased. This seemed greater in cells grown with maltose than with glucose but the latter cells also synthesised distinct amounts of waxes. The total quantity of lipid per cell did not change significantly during loss of viability although the composition of the lipid fraction did.     

The partial oxidation of ethane to acetaldehyde

The mixed oxide (B---P---O) prepared from H₃BO₃ and P₂O₅ was modified to a selective catalyst (B---P---O(S)) to the formation of CH₃CHO after prolonged treatment in a gas mixture of reactants. The contribution of gas-phase chain reactionsto the catalytic oxidations of C₂H₆ on the B---P---O(S) could be neglected. The catalytic active species on the B---P---O(S) was suggested to be a highly dispersed boron oxide supported on BPO₄ crystals. The mixed oxide prepared from BPO₄, H₃BO₃(7atom%), and P₂O₅(3atom%) showed the highest yield of CH₃CHO.