Partitioning of predominant lipophilic bioactives (squalene, α-tocopherol and β-sitosterol) during olive oil processing

The partitioning of predominant lipophilic olive bioactives (squalene, β-sitosterol and α-tocopherol) in olive oil processing was studied for the first time using common integrated olive milling plant. 7% of the oil and 5% of the squalene present in the olives was lost in the last pomace. β-sitosterol and α-tocopherol, which are mainly concentrated in the seed of the olive fruit, was recovered in virgin olive oil to a lesser extent (66%, 67%) while 10% were lost in last pomace. Thus, the loss of β-sitosterol and α-tocopherol was higher than that of squalene and oil, due to nonrecovered bound forms. 19% of the α-tocopherol and 21% of the β-sitosterol loss was unaccounted for, which can be attributed to degradation of α-tocopherol and incomplete recovery of sterols from the olive and pomace matrices. This study provides a basis for process development studies.     

Engineering of Saccharomyces cerevisiae for the production of (+)-ambrein

The triterpenoid (+)-ambrein is the major component of ambergris, a coprolite of the sperm whale that can only be rarely found on shores. Upon oxidative degradation of (+)-ambrein, several fragrance molecules are formed, amongst them (−)-ambrox, one of the highest valued compounds in the perfume industry. In order to generate a Saccharomyces cerevisiae whole-cell biocatalyst for the production of (+)-ambrein, intracellular supply of the squalene was enhanced by overexpression of two central enzymes in the mevalonate and sterol biosynthesis pathway, namely the N-terminally truncated 3-hydroxy-3-methylglutaryl-CoA reductase 1 (tHMG) and the squalene synthase (ERG9). In addition, another key enzyme in sterol biosynthesis, squalene epoxidase (ERG1) was inhibited by an experimentally defined amount of the inhibitor terbinafine in order to reduce flux of squalene towards ergosterol biosynthesis while retaining sufficient activity to maintain cell viability and growth. Heterologous expression of a promiscuous variant of Bacillus megaterium tetraprenyl-β-curcumene cyclase (BmeTC-D373C), which has been shown to be able to catalyse the conversion of squalene to 3-deoxyachillol and then further to (+)-ambrein resulted in production of these triterpenoids in S. cerevisiae for the first time. Triterpenoid yields are comparable with the best microbial production chassis described in literature so far, the methylotrophic yeast Pichia pastoris. Consequently, we discuss similarities and differences of these two yeast species when applied for whole-cell (+)-ambrein production.     

Associations between Oil Yield and Profile of Fatty Acids,Sterols, Squalene,Carotenoids, and Tocopherols in Seed Oil of Selected Japanese Quince Genotypes during Fruit Development

Tocopherols, phytosterols, carotenoids, and squalene are present in mature seeds of Japanese quince. Yet, little is known about the relationship between these compounds and oil yield during fruit and seed development. The profile change of lipophilic compounds during fruit and seed development in Japanese quince cultivars “Darius,” “Rondo,” and “Rasa” is investigated. It is shown here that during fruit and seed development, there is a significant reduction, three‐ to over tenfold, in the concentration of minor bioactive compounds in seed oil. It is recorded that delay between synthesis of tocopherols and oil in Japanese quince seeds during the fruit development results in a logarithmic relationship between the oil content and tocopherols concentration in the seed oil (R² = 0.980). Similar trends are observed between oil yield and phytosterols, and carotenoids (R² = 0.927 and R² = 0.959, respectively). The profile of fatty acids during the development of the seeds significantly is changed. The reduction of linoleic, palmitic, and gondoic acids levels and increment of oleic acid is noted. The oil content, profile of fatty acids, and concentration of bioactive compounds in all three genotypes of Japanese quince do not change significantly statistically during the last month of fruit development. Practical Applications: Some fruits are harvested at different degrees of maturity mainly due to a logistic issue and uneven ripening of fruits, which affects the chemical composition of whole fruit including seeds. Therefore, it would be good to know how the chemical composition is changing in plant material during development especially in the last month before harvest. Production of Japanese quince continues to rise year to year and with it the volume of generated by‐products such as seeds. This study demonstrates how it changes the oil content, profile of fatty acid, and concentration of tocopherols, squalene, phytosterols, and carotenoids in the seeds and seed oil of three Japanese quince cultivars “Rondo,” “Darius,” and “Rasa” during plant development. The provided information can be very useful for the manufactories oriented on the processing of by‐products, mainly seeds, generated by other branches of industry, for instance, fruit‐processing.     

Synthesis,Computational Studies and Assessment of in Vitro Activity of Squalene Derivatives as Carbonic Anhydrase Inhibitors

We report novel molecules incorporating the nontoxic squalene scaffold and different carbonic anhydrase inhibitors (CAIs). Potent inhibitory action, in the low-nanomolar range, was detected against isoforms hCA II for sulfonamide derivatives, which proved to be selective against this isoform over the tumor-associate hCA IX and XII isoforms. On the other hand, coumarin derivatives showed weak potency but high selectivity against the tumor-associated isoform CA IX. These compounds are interesting candidates for preclinical evaluation in glaucoma or various tumors in which the two enzymes are involved. In addition, an in silico study of inhibitor-bound hCA II revealed extensive interactions with the hydrophobic pocket of the active site and provided molecular insights into the binding properties of these new inhibitors.     

甜瓜种子性状及甜瓜籽油中角鲨烯含量研究

采用气相色谱法等对黄色马泡瓜、包瓜等30个甜瓜品种角鲨烯含量进行测定。种子形态特征及相关性分析得知百粒重与面积比和角鲨烯含量均成极显著相关水平,但百粒重与面积比的极显著关系为负相关,即面积比越大,百粒重越小,而角鲨烯含量却随着百粒重的增加有所增加。这30个品种甜瓜籽油中角鲨烯的含量较高(641.7~2 858.3 mg/kg),经主成分分析表明,共有4个主成分对其贡献率较大。在此基础上进行聚类分析可以将其分为4类,其中第一、二类群,面积比和单个重量较小,但角鲨烯含量较高,第三、四类群则长/宽均值较大,角鲨烯含量较高。     

Capillary supercritical fluid chromatographic analysis of shark liver oils

The liver oils of six different shark species have been analyzed by supercritical fluid chromatography (SFC). The liver oils were from the species Pseudotriakis microdon (False catshark), Centroscymnus coelepsis (Portuguese dogfish), Centrophorus squanosus (Leafscalp gulper shark), Deanea calceus (Birdbeak dogfish), Etmopterus princips (Greater lantern shark), and Centroscymnus crepidater (Longnose velvet dogfish). The method was capable of direct quantitation of squalene and cholesterol, while quantitation of triacylglycerols, cholesterol esters, and diacylglycerol ethers required thin-layer chromatographic fractionation prior to SFC analysis. The iodine values of the liver oil samples gave a linear correlation when plotted against the squalene content found by SFC. The variation of squalene content within one shark species is large, and there are large differences in squalene content from species to species. The squalene contents varied between 0.22 and 82.54 wt%. The identity of the glycerol ethers was investigated by SFC of the unsaponifiable matter. The major glycerol ethers contained chimyl, batyl, and selachyl alcohol.     

提取方法对茶油中活性成分角鲨烯含量的影响

采用不同的工艺和溶剂从油茶籽中提取茶油,并对经甲酯化处理的茶油进行气相色谱分析、GC-MS联用分析、标准物质共同色谱比对等方法,研究了不同的提取方法对茶油中生物活性物质角鲨烯含量的影响。结果表明,茶油中含有比较多的角鲨烯,其含量随提取方法而不同,直接压榨法、己烷提取法、石油醚提取法、丙酮提取法和三氯甲烷提取法等所得到的茶油中角鲨烯的含量分别为2.98%、7.62%、0.94%、0.29%和0.54%。     

A Novel Function of the Triterpene Squalene in a Tritrophic System

Changes in apple leaf chemistry after infestation by leafminers and their effect on both host location and host habitat location of the generalist parasitoid Pholetesor bicolor were investigated. Chemical analysis of leaf solvent extracts from healthy and leafminer-damaged leaves revealed that herbivory increased the amount of the triterpene squalene (C₃₀H₅₀), whereas quantities of all other identified compounds were similar in both plant treatments. To assess the response of parasitoids to host location cues, contact bioassays were conducted with naïve females. Results showed that parasitoids performed a characteristic ovipositional probing more often on the mine-damaged than on the healthy leaf. This behavior was triggered by a hexane extract of the mine-damaged leaf, but not by a healthy leaf extract. A synthetic mixture of the compounds identified in the extract triggered a similar response. A mixture devoid of squalene was not active, whereas squalene alone elicited the probing behavior. To assess the use of the identified compounds in habitat location, Y-tube olfactometer experiments were conducted with naïve and experienced females. Results showed that squalene is not involved in habitat location and has no priming effect on P. bicolor. While other triterpenes are known to mediate habitat location of parasitoids, this is the first report in which a plant triterpene is shown to mediate host location of a parasitoid. The biological and ecological functions of squalene on all three trophic levels are discussed.     

Extraction of Teucrium manghuaense and Evaluation of the Bioactivity of Its Extract

The ethanol extract of Teucrium manghuaense grown in Hainan province (China) was analysed by GC and GC/MS. Of the constituents 84–96% were identified on the basis of their GC retention times and their mass spectra in regard to authentic compounds. The results revealed that it contained 9-methyl-9-azabicyclo[4.2.1]nonane (7.43%), 2-methylpyrrolidine (19.42%), 3,7,11,15-tetramethyl-2-hexadecen-1-ol (10.84%), and squalene (28.55%), as major components, constituting 66.24% of the extract. The chemical characterization of the ethanol extract by GC/MS also allowed identification of 1-octen-3-one (3.41%), 2-pentyl-piperidine (2.25%), 1-(2-methyl-1-propenyl)-piperidine (4.63%), 2,2′-diethoxy-5,5′-bi-1-pyrroline (0.41%), (Z,Z,Z)-9,12,15-octadecatrieniic acid, 2,3-dihydroxypropyl ester (1.56%), vitamin E (2.95%) and stigmasterol (1.39%). Finally, the antioxidant and anti-tumor activities of the ethanol extract have been evaluated. Results show that the extract of Teucrium manghuaense leaf possesses strong DPPH⁻, hydroxyl radical scavenging activity and anti-tumor activity.