Hybrid <Emphasis Type="Italic">Hibiscus</Emphasis> seed oil compositions

The seeds of cultivated Hibiscus spp. were extracted with supercritical carbon dioxide, and the resulting extracts were analyzed to identify the major TG components as the corresponding FAME. The seed oils were composed predominantly of oleic and linoleic FA (69.6–83.4%) with lesser amounts of palmitic acid (14.8–27.0%). Minor amounts of C14, C18, and C20 saturated FA were also detected. The oil content of the seeds was determined to be between 11.8 and 22.1 wt% for hybrid varieties and between 8.9 and 29.5 wt% for the native species from which the hybrid varieties were developed. The protein content of the defatted seed meal averaged 20% for the hybrid varieties. The composition of the extracted hibiscus seed oils suggests potential edible applications.     

Fatty acids of the seeds of <Emphasis Type="Italic">Origanum onites</Emphasis> L. and <Emphasis Type="Italic">O. vulgare</Emphasis> L.

Seed oils of Origanum onites L. from the Antalya and Mugla regions and O. vulgare L. from the Kirklareli region of Turkey were extracted with hexane in a Soxhlet apparatus. The oil yields were 14.1–20.0 and 18.5%, respectively. FA compositions of the seed oils were determined by GC and GC/MS. Twenty FA were identified in both O. onites and O. vulgare seeds. The major FA of both species were linolenic (56.3–57.0%; 61.8%), linoleic (21.5–21.7%; 18.8%), oleic (8.7–8.9%; 5.9%), palmitic (5.9–6.5%; 5.5%), stearic (2.1–2.4%; 2.1%), and (Z)-11-octadecenoic (0.6–0.8%; 0.5%), respectively.     

Fatty Acid Composition and Oil Yield in Fruits of Five <Emphasis Type="Italic">Arecaceae</Emphasis> Species Grown in Cuba

The present study targeted the whole-fruit oil yield and fatty acid composition from five of the most abundant Arecaceae species grown in Cuba. The oil yields (% dry weight), determined by the Soxhlet extraction technique with hexane, were 25.5, 5.3, 6.9, 5.4, and 6.4% for Roystonea regia, Colpothrinax wrightii, Sabal maritima, Sabal palmetto and Thrinax radiata, respectively. The free fatty acid (FFA) content varied from 2.7 to 6.8%. Fatty acid (FA) profiles of the oils indicated that lauric acid (13.7–44.4%), myristic acid (9.4–22.4%) and palmitic acid (9.2–17.1%) as major saturated FA; whereas oleic acid (9.6–42.7%) and linoleic acid (9.3–17.0%) as major unsaturated FA. R. regia fruit seemed the most promising among Arecaceae grown in Cuba because of its high oil yield and low oil FFA content.     

A comparative study of fatty acid profiles of <Emphasis Type="Italic">Passiflora</Emphasis> seed oils from Uganda

A comparative study is presented of the FA composition (FAC) of the seed oils from the yellow passion fruit Passiflora edulis Sims var. flavicarpa (I), the purple fruit Passiflora edulis Sims var. edulis (II), the purple Kawanda hybrid, which is a cross between I and II (III), and the light-yellow apple passion fruit Passiflora maliformis L. (IV) grown in Uganda. Oil yields from the four varieties were between 18.5 and 28.3%. A GC analysis of the oils showed the most dominant FA to be linoleic (67.8–74.3%), oleic (13.6–16.9%), palmitic (8.8–11.0%), stearic (2.2–3.1%), and α-linolenic (0.3–0.4%) acids. The unsaturated FA content in the oils was high (85.4–88.6%). Iodine values of the seed oils of I, II, III, and IV calculated from the FAC were 133, 141, 133, and 138, respectively. The FAC and the iodine value of the seed oil in III are distinctly closer to the rootstock (I) than the scion (II). This indicates that the rootstock influence on the FAC of passion fruit seeds is graft-transmissible. The study further confirms that passion fruit seed oils represent a good source of essential unsaturated FA.     

Fatty Acid Composition of Turkish <Emphasis Type="Italic">Rhododendron</Emphasis> Species

This study describes work aimed at the rapid evaluation of the fatty acid (FA) composition of Turkish Rhododendron species, particularly the leaves and the flowers of the toxic plants, R. ponticum and R. luteum. The FA profiles of the available parts of three other nonpoisonous Rhododendron species were also investigated. Subtotal extracts obtained (using n-hexane, chloroform and methanol) from total chloroform:methanol (1:1) extracts were analyzed and compared to each other. Palmitic acid was found to be the most abundant FA in almost all Rhododendron extracts, and the majority of leaf and flower extracts contained significant portions of C18 unsaturated FAs (18:1n-9, 18:2n-6, 18:3n-3). The n-hexane extracts of R. ponticum leaves and R. luteum flowers were unique, as they contained an unusual series of even-chain iso FAs (C16–C24). Especially the n-hexane extracts were found to comprise uncommon FAs with odd-numbered carbons (C13–C29). Overall, n-hexane proved to be the best solvent by representing the richest FA profile, whereas chloroform or methanol appeared less suitable for FA analyses. Appreciable intra-species variations in FA compositions among the leaves as well as other anatomical parts examined were observed. This study highlights the chemotaxonomical importance of the FAs for the genus Rhododendron.     

Interprovenance variation in the composition of <Emphasis Type="Italic">Moringa oleifera</Emphasis> oilseeds from Pakistan

Interprovenance variation was examined in the composition of Moringa oleifera oilseeds from Pakistan. The hexane-extracted oil content of M. oleifera seeds harvested in the vicinity of the University of Agriculture, Faisalabad (Punjab, Pakistan), Bahauddin Zakariya University (Multan, Pakistan), and the University of Sindh, Jamshoro (Sindh, Pakistan), ranged from 33.23 to 40.90%. Protein, fiber, moisture, and ash contents were found to be 28.52–34.00, 6.52–7.50, 5.90–7.00, and 6.52–7.50%, respectively. The physical and chemical parameters of the extracted M. oleifera oils were as follows: iodine value, 67.20–71.00; refractive index (40°C), 1.4570–1.4637; density (24°C), 0.9012–0.9052 mg/mL; saponification value, 177.29–184.10; unsaponifiable matter, 0.60–0.83%; color (1-in. cell), 1.00–1.50 R+20.00–30.00Y; smoke point, 198–202°C; and acidity (% as oleic acid), 0.50–0.74. Tocopherols (α, γ, and δ) accounted for 114.50–140.42, 58.05–86.70, and 54.20–75.16 mg/kg, respectively, of the oils. The induction periods (Rancimat, 20 L/h, 120°C) of the crude oils were 9.64–10.66 h and were reduced to 8.29–9.10 h after degumming. Specific extinctions at 232 and 270 nm were 1.80–2.50 and 0.54–1.00, respectively. The major sterol fractions of the oils were campesterol (14.13–17.00%), stigmasterol (15.88–19.00%), β-sitosterol (45.30–53.20%), and ͤ⁵-avenasterol (8.84, 11.05%). The Moringa oils were found to contain high levels of oleic acid (up to 76.00%), followed by palmitic, stearic, behenic, and arachidic acids up to levels of 6.54, 6.00, 7.00, and 4.00%, respectively. Most of the parameters of M. oleifera oils indigenous to different agroclimatic regions of Pakistan were comparable to those of typical Moringa seed oils reported in the literature. The results of the present analytical study, compared with those for different vegetable oils, showed M. oleifera to be a potentially valuable oilseed crop.     

Acorn (<Emphasis Type="Italic">Quercus</Emphasis> spp.) fruit lipids: Saponifiable and unsaponifiable fractions: A detailed study

The composition of the oils extracted from the acorn fruit of three species of Mediterranean oaks, Quercus ilex L., Q. suber L., and Q. faginea L., was characterized. Both major and minor components, including FA, TG, sterols, methyl sterols, triterpenic and aliphatic alcohols, tocopherols, and hydrocarbons, were identified by standard methods and MS. High-resolution GLC and HPLC were used for quantification. The FA profile, together with the equivalent carbon numbers and TG carbon numbers, was compared with data for other edible vegetable oils. Oil yield, expressed as wet weight, was 5% (w/w). Sterol content was remarkable for the three species (8,563–11,420 mg/kg), with β-sitosterol being the most abundant (80%). Oils were also high in tocopherol, with a wide variation between species (165–456 mg/kg) but with γ-tocopherol predominating in all three oils (90% of the total tocopherol content). Also, high terpenic alcohol contents were found (1527–2984 mg/kg), with dammaradienol and β-amyrin being the most abundant (33–60% of the total alcohol content). Bioactive properties and industrial applications of this underutilized native product are also discussed.     

Use of <Emphasis Type="Italic">Rhizopus delemar</Emphasis> lipase as compared with other lipases for determination of <Emphasis Type="Italic">sn</Emphasis>-2 fatty acids in triacylglycerol

The FA composition in the sn-2 position of TAG is routinely determined after porcine pancreatic lipase hydrolysis. However, the content of saturated FA increased when a pancreatic lipase preparation with higher specific activity was used. Lipase from Rhizopus delemar was selected as a potential replacement lipase for the following reasons: (i) The FA specificity is nearly equivalent in hydrolysis activity toward FA such as lauric, myristic, palmitic, palmitoleic, stearic, oleic, linoleic, and α-linolenic acids; and (ii) lipase from R. delemar hydrolyzes fatty acyl residues at the sn-1,3 positions of TAG. Acyl migration products were present at less than 0.8% in lipase hydrolysates containing 6–14% of sn-2 MAG. A reproducibility CV of less than 5% was obtained in a collaborative study in which the compositions of the main FA at the sn-2 position in olive oil were determined using lipase from R. delemar. This article was presented in part at the Biocatalysis Symposium, 94th AOCS Annual Meeting & Expo, Kansas City, Missouri, May 2003.     

Natural estolides produced by <Emphasis Type="Italic">Pseudomonas</Emphasis> sp. 42A2 grown on oleic acid: Production and characterization

Estolides are a group of FA polyesters resulting from ester bond formation between a hydroxyl or olefinic group of one FA and the terminal carboxyl group of a second FA. These products are commonly found in trace amounts, forming tetraglycerides in several oil seed plants, and have been produced by acid clay and enzymatic catalysis in vitro. In this study, natural estolides produced by a bacterial culture are presented for the first time. Pseudomonas sp. 42A2 produced (E)-10-hydroxy-8-octadecenoic acid and (E)-7,10-dihydroxy-8-octadecenoic acid when grown on oleic acid. It is suggested that these FA were polymerized in culture by a lipase produced by the bacterial strain, resulting in a mixture of estolides. These compounds amounted to 3.8 g/L after 72 h of incubation. LC-MS analysis indicated that the types of estolides formed were dimers (m/z 560–610), trimers (m/z 845–906), tetramers (m/z 1122–1202), pentamers (m/z 1328–1424), and hexamers (m/z 1554–1788), with a relative abundance of 27.5, 19.4, 15, 9.7, and 11%, respectively. This is the first report in which hexamers were detected in a bacterial culture.     

Characterization and compositional studies of the oils from some legume cultivars, <Emphasis Type="Italic">Phaseolus vulgaris</Emphasis>, grown in Southern Africa

Seed oils from six legume cultivars of Phaseolus vulgaris, grown in the Kingdom of Lesotho, were extracted and their physicochemical properties and FA compositions were determined in order to compare their dietary lipids with those in P. vulgaris cultivars grown in other parts of the world. The oil content of the beans was very low, ranging from 1.5 to 2.0% (w/w). The acid values ranged from 11.0 to 19.2 mg KOH/g, whereas a combination of the PV and the p-anisidine values in Holm's equation gave oxidation values that ranged from 11.0 to 15.0. Thus, considerable enzymatic hydrolysis and oxidation had taken place in the beans during storage. Iodine values ranged from 80.5 to 92.3 (Wijs method), indicating moderate unsaturation in the oils. However, capillary GC analysis, supported by proton NMR analysis of the FAME, gave a total unsaturation range from 79.67 to 84.24%. The dominant FA were α-linolenic acid (36.47–48.81%) and linoleic acid (20.96–36.10%), with appreciable amounts of palmitic acid (14.33–18.23%). This FA composition pattern is quite similar to the FA distribution reported for low oil-bearing legume seeds. Thus, notwithstanding the different climatic and soil conditions, the general properties of lipids in the southern African legume cultivars were quite similar to those of lipids in P. vulgaris cultivars grown in other parts of the world. The high content of α-linolenic acid in the cultivars of P. vulgaris could very likely play a beneficial role in reducing the risk of coronary heart disease among the large populations consuming them in the southern African region.     

Lipid Classes,Fatty Acid Distributions and Triacylglycerol Molecular Species of Broad Beans (<Emphasis Type="Italic">Vicia faba</Emphasis>)

Seed oils from four legume cultivars of Vicia faba, grown in Japan, were extracted and classified by thin-layer chromatography (TLC) into eight fractions. The major lipid components were triacylglycerols (TAG: 48.8–50.1%) and phospholipids (PL: 47.5–50.5%), while hydrocarbons (HC), steryl esters (SE), free fatty acids (FFA), diacylglycerols (1,3- and 1,2-DAG) and monoacylglycerols (MAG) were present in minor proportions (1.8–2.4%). All lipid samples had high amounts of total unsaturated FA, representing 79.7–82.8% and 77.6–79.7% for TAG and PL, respectively. Molecular species and FA distributions of TAG, isolated from the total lipids in the broad beans, were analyzed by a combination of argentation-TLC and GC. Fourteen different molecular species were detected. With a few exceptions, the main TAG components were S₂D (6.1–8.9%), SD₂ (7.8–10.5%), SMT (6.3–8.5%), M₂D (4.5–6.2%), MD₂ (18.9–21.8%), D₃ (21.0–23.9%) and MDT (8.1–10.2%) (where S, M, D, and T denote a saturated fatty acid, a monoene, a diene, and a triene, respectively). These results suggest that the lipid classes, FA distributions and TAG molecular species of broad beans are not dependent on the cultivation areas during the growing season.     

Reaction selectivity of <Emphasis Type="Italic">Burkholderia cepacia</Emphasis> (PS-30) lipase as influenced by monoacylation of <Emphasis Type="Italic">sn</Emphasis>-glycerol

Reaction selectivities were determined in multicompetitive reactions mediated by Burkholderia cepacia lipase (Amano PS-30) at a water activity of 0.19 in hexane. Saturated FA (C4–C18 even chain) and oleic acid (C18∶1) were reacted with a single alcohol, glycerol, α-or β-MAG, containing C4, C10, C16, or C18∶1 individually as alcohol cosubstrate. Similar odrinal patterns of FA selectivity, with C8, C10, and C16 preferred over others, were generally observed for incorporation of FA into specific acylglycerol (AG) pools of the 24 specific cases evaluated. The three exceptions were enrichment of C14 and C18 in the MAG pool with α-C16-MAG, substrate, and a general suppression of >C8 incorporation into the TAG pool for reactions with α-C10- and α-C16-MAG. PS-30 lipase selectivity toward MAG was in descending order: α/β-C4-MAG>β-C10-MAG>β-C16-MAG>α/β-C18∶1-MAG>α-C10-MAG>α-C16-MAG. Selectivity in channeling CX of the original CX-MAG substrates into higher AG species was in descending order: α-C10-MAG∼α-C16-MAG>α-C18∶1-MAG>β-C10-MAG∼β-C16-MAG∼β-C18∶1-MAG >α/β-C4-MAG. Generally, MAG were better acyl donors than FA for esterification reactions leading to DAG formation. These observations are relevant to the design of biocatalytic processes intended to yield specifically structured TAG.     

Chemical characterization of oils and meals from wild sunflower (<Emphasis Type="Italic">helianthus petiolaris</Emphasis> nutt)

Chemical characterizations of oils and meals from the wild sunflower species Helianthus petiolaris Nutt and their comparison with those from cultivated sunflower (H. annuus) were performed. Seeds from indigenous populations of H. petiolaris were harvested in Argentina in different years. The analytical parameters studied were as follows: (i) FA profile, PV, p-anisidine value, oxidative stability, phosphorus and phospholipid content, tocopherols, polar compounds, and waxes in the extracted oils; and (ii) moisture, ash, crude fiber, metals, sugars, urease activity, starch, protein, available lysine, neutral detergent fiber, acid detergent fiber, lignin, gross energy, and amino acid content in the residual meals. The products from wild sunflower seed, which yielded 27–30% oil by solvent extraction, showed some characteristics similar to the commercial products. Nevertheless, the oil had lower quality and stability owing to the high unsaturation levels and lower concentrations of antioxidant components, and the meal had a lower protein content. The phospholipid content was significantly lower than in industrial crude sunflower oils. Most of the important parameters in the meal such as available lysine, gross energy, and digestibility compared favorably with those for cultivated sunflower meals. The results showed the potential for using these meals for animal feed.     

Profiles of Lipid Components,Fatty Acid Compositions and Triacylglycerol Molecular Species of Adzuki Beans (<Emphasis Type="Italic">Vigna angularis</Emphasis>)

Fatty acid (FA) compositions and molecular species of triacylglycerols (TAG) isolated from total lipids extracted from adzuki beans (Vigna angularis) were determined with a combination of AgNO₃-TLC and GC, and were compared in relation to the content of endogenous antioxidants analyzed by HPLC. δ-Tocopherol was present in the highest concentration (53.7–89.3 mg/kg), and γ-tocopherol in small amounts (11.2–14.8 mg/kg). The main lipid components were phospholipids (72.2–73.4%) and TAG (20.6–21.9%), whilst other components were also present in minor proportions (0.1–3.4%). Eighteen different TAG molecular species were identified and quantified by successive applications of AgNO₃-TLC and GC. The main components were SMD (4.6–5.0%), S₂T (13.4–16.4%), SD₂ (11.8–14.3%), SMT (7.3–8.3%), SDT (9.9–10.6%), D₃ (6.9–7.9%), MT₂ (5.2–6.3%), D₂T (7.0–11.2%), DT₂ (7.4–7.6%) and T₃ (6.2–7.2%) (where S, M, D, and T denote a saturated FA, a monoene, a diene, and a triene, respectively). No marked difference (P > 0.05) in the molecular species composition could be observed among the five cultivars. The results could be useful to both consumers and producers for manufacturing traditional adzuki confectionaries in Japan and elsewhere.     

Fatty acid (FA) composition and contents of <Emphasis Type="Italic">trans</Emphasis> unsaturated FA in hydrogenated vegetable oils and blended fats from Pakistan

This study presents the FA composition and trans FA (TFA) contents of different hydrogenated vegetable oils and blended fats marketed in Pakistan. Thirty-four vanaspati (vegetable ghee), 11 shortenings, and 11 margarines were analyzed. The contents of saturated FA, cis monounsaturated FA, and cis PUFA were in the following ranges: vanaspati 27.8–49.5, 22.2–27.5, 9.3–13.1%; vegetable shortenings 37.1–55.5, 15.8–36.0, 2.7–7.0%; and margarines 44.2–55.8, 21.7–39.9, 2.9–20.5%, respectively. Results showed significantly higher amounts of TFA in vanaspati samples, from 14.2 to 34.3%. Shortenings contained TFA proportions of 7.3–31.7%. The contents of TFA in hard-type margarines were in the range of 1.6–23.1%, whereas soft margarines contained less than 4.1% TFA.     

<Emphasis Type="Italic">Trans</Emphasis> FA in sunflower oil at different steps of refining

The contents of total trans FA of sunflower oils at different stages of refining processes were determined by capillary GLC. The contents of 18∶1, 18∶2, and 18∶3 trans acids were 0.22±0.03, 2.31±0.23, and 0.03±0.01%, respectively, in physically refined sunflower oils, and 0.05±0.01, 0.69±0.26, and 0.02±0.01%, respectively, in chemically refined sunflower oils. The total trans FA contents drastically increased at the end of the physical refining process. The total trans FA contents of chemically refined sunflower oils were <1%. Because of the high temperature applied in the last stage of physical refining, the content of total trans FA was higher than in chemically refined sunflower oils. The last-stage conditions should be carefully evaluated to reduce the formation of trans FA during physical refining.     

Isomers of hexadecenoic and hexadecadienoic acids in <Emphasis Type="Italic">Androsace septentrionalis</Emphasis> (Primulaceae) seed oil

Seeds of Androsace septentrionalis of the genus Androsace (tribus Primuleae) from the plant family Primulaceae were studied for their oil content and FA composition. The seed oil of A. septentrionalis was found to contain two unusual FA rarely occurring in plants: 11-cis-hexadecenoic acid (16∶1Δ11c or 16∶1n−5) and 9-cis,12-cis-hexadecadienoic acid (16∶2Δ9c,12c or 16∶2n−4). It also contained an unusually high amount (21.4%) of 9-cis-hexadecenoic acid (palmitoleic acid; 16∶1Δ9c or 16∶1n−7), i.e., at a level higher than that of oleic acid, in addition to common FA. Compared with most plant seed oils, at 3.8% the level of 18∶1Δ11c (or 18∶1n−7) also was elevated. The nonidentity of the Androsace 16∶2-acid with the 16∶2-acid, which is very typical for Ranunculus spp., as well as its identity with the 16∶2-acid typically found in Asclepiadaceae was established by co-chromatography. The structure and composition of the constituent FA of A. septentrionalis were also determined by various chromatographic methods (TLC, Ag⁺-TLC, capillary GLC) and spectroscopic methods (IR, GC-MS). The significant deviation of the Androsace FA pattern from that of other Primuleae, indicating a separate phylogenetic position of Androsace, is discussed.     

Cyanolipid-rich seed oils from <Emphasis Type="Italic">Allophylus natalensis</Emphasis> and <Emphasis Type="Italic">A. dregeanus</Emphasis>

As a continuation of our study on plants of the Sapindaceae, the chemical composition of the oil extracted from seeds of Allophylus natalensis (Sonder) De Winter and of A. dregeanus (Sonder) De Winter has been investigated. The oil from both species contained approximately equal amounts of TAG and type I cyanolipids (CL), 1-cyano-2-hydroxymethylprop-2-en-1-oldiesters, with minor amounts of type III CL, 1-cyano-2-hydroxymethylprop-1-en-3-ol-diesters. Structural investigation of the oil components was accomplished by chemical, chromatographic (TLC, CC, GC, and GC-MS), and spectroscopic (IR, NMR) means. GC and GC-MS analysis showed that C₂₀ FA were dominant in the CL components of the oil from the two species (44–80% vs. 21–26% in TAG), with cis-11-eicosenoic acid (36–46%) and cis 13-eicosenoic acid (paullinic acid, 23–37%) as the major esterified fatty acyl chains in A. natalensis and A. dregeanus, respectively. cis-Vaccenic acid was particularly abundant (11–31%) in the CL from A. dregeanus, whereas eicosanoic acid (10–22%) was also a major component of CL in both species.     

Effects of Geographical Origin on the Conjugated Linolenic Acid of <Emphasis Type="Italic">Trichosanthes kirilowii</Emphasis> Maxim Seed Oil

Trichosanthes kirilowii Maxim (T. kirilowii) seeds from four geographical locations (Changxing, Quzhou, Yuexi, Dongzhi) contained 26.15–49.41% oil and 28.68–37.90% protein. The seed oil was distinguished by the conjugated linolenic acids, punicic acid (PA) and α-eleostearic acid (α-ESA). The main fatty acids in T. kirilowii seed oils were ranked in the following order: punicic acid (33.09–39.15%), linolenic acid (33.77–38.66%), oleic acid (15.15–24.88%), palmitic acid (2.36–4.86%). PA was the main isomer of CLNA (33.09–39.15%). No significant differences were found either in PA content or in α-ESA content of T. kirilowii seed from these geographical locations. Little difference was observed in the quantitative composition of the lipid contents of seeds from different geographical locations. The α-tocopherol content of T. kirilowii seed ranged from 6.34 to 31.74 mg/100 g, with the highest levels in Changxing seeds. The present results showed that T. kirilowii seeds were especially rich in PA, and their contents were not influenced by the geographical locations. Variation in some proximate compositions by geographical locations may be caused by ecological conditions, temperature, climate condition, technical and cultural conditions.     

Biosynthesis of tetrahydrofuranyl fatty acids from linoleic acid by <Emphasis Type="Italic">clavibacter</Emphasis> sp. ALA2

Clavibacter sp. ALA2 converts linoleic acid into many novel oxygenated products including hydroxy FA and tetrahydrofuranyl unsaturated FA (THFA). One of them was tentatively identified by GC-MS as 12,13,16-trihydroxy-9(Z)-octadecenoic acid (12,13,16-THOA) (Hou, C.T., H.W. Gardner, and W. Brown, J Am. Oil Chem. Soc. 78∶1167–1169, 2001). We have separated and purified 12,13,16-THOA from its isomer, 12,13,17-THOA, by silica gel column chromatography and by preparative TLC. Its structure was then confirmed by proton and ¹³C NMR analyses. Purified 12,13,16-THOA was used as a substrate to study the biosynthesis of THFA. Within 24 h of incubation, cells of strain ALA2 converted 12,13,16-THOA to both 12-hydroxy-13,16-epoxy-9(Z)-octadecenoic acid (12-hydroxy-THFA) and 7,12-dihydroxy-13,16-epoxy-9(Z)-octadecenoic acid (7,12-dihydroxy-THFA). The relative abundance of 7,12-dihydroxy-THFA increased with incubation time, whereas that of 12,13,16-THOA and of 12-hydroxy-THFA decreased. Therefore, the biosynthetic pathway of THFA from linoleic acid by strain ALA2 is as follows: linoleic acid→12,13-dihydroxy-9(Z)-octadecenoic acid→12,13,16-THOA→12-hydroxy-THEA→7,12-dihydroxy-THFA.