5,8‐Dihydroxy‐9,12,15(Z,Z,Z)‐Octadecatrienoic Acid Production by Recombinant Cells Expressing Aspergillus nidulans Diol Synthase

Whole cells of recombinant Escherichia coli expressing diol synthase from Aspergillus nidulans produced 5,8‐dihydroxy‐9,12,15(Z,Z,Z)‐octadecatrienoic acid from α‐linolenic acid via 8‐hydroperoxy‐9,12,15(Z,Z,Z)‐octadecatrienoic acid as an intermediate. The optimal conditions for 5,8‐dihydroxy‐9,12,15(Z,Z,Z)‐octadecatrienoic acid production using whole recombinant cells were exhibited at pH 7.0, 40 °C, and 250 rpm with 40 g/L cells, 12 g/L, α‐linolenic acid, and 5 % (v/v) dimethyl sulfoxide in a 250‐mL baffled flask containing 50 mL reaction solution. Under these conditions, whole recombinant cells produced 9.1 g/L 5,8‐dihydroxy‐9,12,15(Z,Z,Z)‐octadecatrienoic acid for 100 min, with a conversion yield of 75 % (w/w), a volumetric productivity of 5.5 g/L/h, and specific productivity of 137 mg/g‐cells/h. As an intermediate, 8‐hydroperoxy‐9,12,15(Z,Z,Z)‐octadecatrienoic acid was observed at approximately 1.4 g/L after 100 min. With regard to dihydroxy fatty acid production, this is the highest reported volumetric and specific productivities thus far. This is the first report on the biotechnological production of 5,8‐dihydroxy‐9,12,15(Z,Z,Z)‐octadecatrienoic acid.     

13‐Oxo‐9(Z),11(E),15(Z)‐octadecatrienoic Acid Activates Peroxisome Proliferator‐Activated Receptor γ in Adipocytes

Peroxisome proliferator‐activated receptor (PPAR)γ is expressed in adipose tissue and plays a key role in the regulation of adipogenesis. PPARγ activators are known to have potent antihyperglycemic activity and are used to treat insulin resistance associated with diabetes. Therefore, many natural and synthetic agonists of PPARγ are used in the treatment of glucose disorders. In the present study, we found that 13‐oxo‐9(Z),11(E),15(Z)‐octadecatrienoic acid (13‐oxo‐OTA), a linolenic acid derivative, is present in the extract of tomato (Solanum lycopersicum), Mandarin orange (Citrus reticulata), and bitter gourd (Momordica charantia). We also found that 13‐oxo‐OTA activated PPARγ and induced the mRNA expression of PPARγ target genes in adipocytes, thereby promoting differentiation. Furthermore, 13‐oxo‐OTA induced secretion of adiponectin and stimulated glucose uptake in adipocytes. To our knowledge, this is the first study to report that 13‐oxo‐OTA induces adipogenesis through PPARγ activation and to present 13‐oxo‐OTA as a valuable food‐derived compound that may be applied in the management of glucose metabolism disorders.     

Responses of MAC‐T Cells to Inhibited Stearoyl‐CoA Desaturase 1 during cis‐9, trans‐11 Conjugated Linoleic Acid Synthesis

Cis‐9‐conjugated, trans‐11‐conjugated linoleic acid (CLA) is known for its positive activities on human health. The synthesis of cis‐9, trans‐11 CLA in mammary glands is generally thought to be catalyzed by stearoyl‐CoA desaturase 1 (SCD1), but this has not been rigorously established. In this study, we hypothesized that the inhibition of SCD1 (by CAY10566) would block the synthesis of cis‐9, trans‐11 CLA in bovine mammary alveolar cells (MAC‐T) cells. Results showed that MAC‐T cells incubated with 10 nM CAY10566 for 12 h (CAY) produced less cis‐9, trans‐11 CLA (p < 0.01), lower 14:1/(14:1 + 14:0)% (p < 0.01), more trans‐11 18:1 (TVA) accumulation (p < 0.01), and reduced SCD1 mRNA levels (p < 0.01) compared with the control group (CON). Moreover, the mRNA abundances of sterol regulatory element‐binding protein 1 [SREBPF1], acyl‐CoA synthetase short‐chain family member 2 [ACSS2], and lipin 1 [LPIN1] were significantly elevated when SCD1 was inhibited in the CAY group (p < 0.05). Taken together, CAY10566 inhibition of SCD1 resulted in lower cis‐9, trans‐11 CLA synthesis ability, and SREBF1, ACSSS2, and LPIN1 were negatively associated with SCD1. These findings not only provide the direct evidence that cis‐9, trans‐11 CLA synthesis is catalyzed by SCD1, but also help us understand the responses of MAC‐T cells to SCD1 inhibition.     

Enantioselective formation of an α, β-epoxy alcohol by reaction of methyl 13(S)-hydroperoxy-9(Z), 11(E)-octadecadienoate with titanium isopropoxide

Methyl 11(R), 12(R)-epoxy-13(S)-hydroxy-9(Z)-octadecenoate (threo isomer) was generated from linoleic acid by the sequential action of an enzyme and two chemical reagents. Linoleic acid was treated with lipoxygenase to yield its corresponding hydroperoxide [13(S)-hydroperoxy-9(Z), 11(E)-octadecadienoic acid]. After methylation with CH₂N₂, the hydroperoxide was treated with titanium (IV) isopropoxide [Ti(O-i-Pr)₄] at 5°C for 1 h. The products were separated by normal-phase high-performance liquid chromatography and characterized with gas chromatography-mass spectrometry, infrared spectroscopy, and nuclear magnetic resonance spectroscopy. Approximately 30% of the product was methyl 13(S)-hydroxy-9(Z), 11(E)-octadecadienoate. Over 60% of the isolated product was methyl 11(R), 12(R)-epoxy-13(S)-hydroxy-9(Z)-octadecenoate. After quenching Ti(O-i-Pr)₄ with water, the spent catalyst could be removed from the fatty products by partitioning between CH₂Cl₂ and water. These results demonstrate that Ti(O-i-Pr)₄ selectively promotes the formation of an α-epoxide with the threo configuration. It was critically important to start with dry methyl 13(S)-hydroperoxy-9(Z),11(E)-octadecadienoate because the presence of small amounts of water in the reaction medium resulted in the complete hydrolysis of epoxy alcohol to trihydroxy products.     

Fibrous long‐chain organic acid cellulose esters and their characterization by diffuse reflectance FTIR spectroscopy,solid‐state CP/MAS 13C‐NMR,and X‐ray diffraction

Unsaturated and saturated organic acids with 11 and 18 carbon atoms, respectively, were used in a heterogeneous esterification reaction in the pyridine/toluene sulfonyl chloride system to prepare fibrous cellulose esters with different degrees of substitution. Highly bleached sulfite cellulose fibers were esterified during a 1‐ or 2‐h reaction time with the following organic acids: undecylenic acid, undecanoic acid, oleic acid, and stearic acid. In all cases, the heterogeneous esterification yielded partially substituted cellulose esters retaining their fibrous structure. The substitution reaction was confirmed by diffuse reflectance infrared spectroscopy and the chemical structures of cellulose esters were identified by solid‐state CP/MAS ¹³C‐NMR (75.3 MHz). X‐ray diffraction analyses showed broadening of the diffraction peaks with a higher degree of substitution of cellulose esters, which suggests structural changes within the cellulose fibers. Because the broadening peaks of X‐ray spectra or the unassigned C‐4 region of substituted cellulose chains in NMR spectra do not allow the calculation of dimensional changes of cellulose crystallites in cellulose esters, the lateral dimensions of crystallites in only cellulose fibers were calculated. The value derived from NMR (4.6 nm) differs by about 11% when compared with the value calculated from X‐ray diffraction data (4.1 nm). © 2000 John Wiley & Sons, Inc. J Appl Polym Sci 78: 1354–1365, 2000     

Short‐Chain Fatty Acids Enhance the Lipid Accumulation of 3T3‐L1 Cells by Modulating the Expression of Enzymes of Fatty Acid Metabolism

Short‐chain fatty acids (SCFA) such as acetic acid, propionic acid, and butyric acid are produced by fermentation by gut microbiota. In this paper, we investigate the effects of SCFA on 3T3‐L1 cells and the underlying molecular mechanisms. The cells were treated with acetic acid, propionic acid, or butyric acid when cells were induced to differentiate into adipocytes. MTT assay was employed to detect the viability of 3T3‐L1 cells. Oil Red O staining was used to visualize the lipid content in 3T3‐L1 cells. A triglyceride assay kit was used to detect the triacylglycerol content in 3T3‐L1 cells. qRT‐PCR and Western blot were used to evaluate the expression of metabolic enzymes. MTT results showed that safe concentrations of acetic acid, propionic acid, and butyric acid were less than 6.4, 3.2, and 0.8 mM, respectively. Oil Red O staining and triacylglycerols detection results showed that treatment with acetic acid, propionic acid, and butyric acid accelerated the 3T3‐L1 adipocyte differentiation. qRT‐PCR and Western blot results showed that the expressions of lipoprotein lipase (LPL), adipocyte fatty acid binding protein 4 (FABP4), fatty acid transporter protein 4 (FATP4), and fatty acid synthase (FAS) were significantly increased by acetic acid, propionic acid, and butyric acid treatment during adipose differentiation (p < 0.05). In conclusion, SCFA promoted lipid accumulation by modulating the expression of enzymes of fatty acid metabolism.