Trihydroxy Fatty Acids Research Articles

Storability and Brewing Quality of Corn Grits: Impacts of Storage Temperature, Particle Size and Key Compositional Factors

Across international brewing operations corn grits from different suppliers vary substantially in terms of their physical properties and chemical composition. Present research aimed to highlight the key factors determining the brewing quality of stored corn grits. Six grits samples were sourced with different particle sizes (that ranged from 0% to 74% of particles ≤0.355 mm) and stored at 10 °C, 20 °C, or 30 °C for 0, 2, 4, 8, 12, and 16 months. Samples were analysed for moisture, extract, lipid, trihydroxy fatty acids (THFA) and pro-oxidant metal ion contents. Particle size range was measured using sieve analysis. Increasing levels of THFA occurred on storage due to lipid oxidation and are negative indicators of brewing quality. PLS-R modelling of THFA levels (R 2 = 0.83) revealed particle size (hence surface area for oxidation) was the most important factor (∼60% of the total R 2) followed by storage time (∼25%) and temperature (∼7%). Metal ion content and lipid content of grits were minor contributors. Results showed that coarser milling of grits offers protection against lipid oxidation even after 8 months at 30 °C. These insights into key factors determining the rate of lipid oxidation in stored corn grits will help to develop better supply chain management and storage practices with which to assure their brewing quality.

Okeanic acid–A, a trihydroxy fatty acid from the Okinawan cyanobacterium Okeania hirsuta

Trihydroxy fatty acids are oxidative metabolites of polyunsaturated fatty acids isolated from plants, bacteria, fungi, and microalgae and have a variety of biological activities. In this study, a new trihydroxy fatty acid, okeanic acid–A (1), was isolated together with malyngic acid (2) and 15,16-dihydromalyngic acid (3) from the cyanobacterium Okeania hirsuta collected in Okinawa, Japan. The planar structure of 1 was elucidated by detailed analyses using high-resolution ESI-MS and 1D and 2D NMR spectroscopy. The absolute configurations of the hydroxy groups in 1 were determined unambiguously by chemical derivatisation and a modified Mosher’s method. These cyanobacterial trihydroxy fatty acids (1–3) have identical configurations at their respective trihydroxy parts. Okeanic acid–A (1) showed mild growth-inhibitory activity against the marine diatom Nitzschia amabilis.

Chemical Constituents of Ulmus minor subsp. minor Fruits Used in the Italian Phytoalimurgic Tradition and Their Anti-inflammatory Activity Evaluation.

The phytochemical investigation of Ulmus minor subsp. minor samaras EtOAc and n-BuOH extracts is reported in this work for the first time, resulting in the isolation and characterization of twenty compounds (1: - 20: ) including one new flavan-3-ol (1: ), one new trihydroxy fatty acid (2: ), and two glycosylated flavonoids (6: - 7: ) whose NMR data are not available in the literature. Structure elucidation of the isolated compounds was obtained by 1D and 2D NMR and HRESIMS data. Prior to further pharmacological investigations, the extracts (100 - 6.25 µg/mL) and compounds 1: - 12: (50 - 5 µM) were tested for their influence on viability of a murine macrophage cell line (J774A.1). Subsequently, extracts and compounds that did not impede viability, were studied for their inhibitory effect on some mediators of inflammation in J774A.1 cells stimulated with lipopolysaccharide of Escherichia coli (LPS). The NO release and the expression of iNOS and COX-2 were then evaluated and both extracts (50 - 6.25 µg/mL) and compounds (20 - 5 µM) significantly inhibited NO release as well as iNOS and COX-2 expression in macrophages. These data highlight the anti-inflammatory properties of several isolated compounds from U. minor samaras supporting their possible alimentary use.

Identification of Non-Volatile Compounds That Impact Flavor Disliking of Whole Wheat Bread Made with Aged Flours.

Whole wheat flour has a shorter shelf life than refined wheat flour due to off-flavor development. An untargeted liquid chromatography/mass spectrometry (LC/MS) flavoromics approach was applied to identify compounds that negatively impact the flavor liking in whole wheat bread made from aged flours. The chemical profiles of thirteen breads made from aged flours were obtained using LC/MS and modeled by orthogonal partial least squares (OPLS) to predict flavor liking. Top predictive chemical features (negatively correlated) were identified as pinellic acid (9S,12S,13S-trihydroxy-10E-octadecenoic acid), 12,13-dihydroxy-9Z-octadecenoic acid, and 1-(9Z,12Z-octadecadienoyl)-sn-glycero-3-phosphocholine. The sensory analysis confirmed the three compounds increased the bitterness intensity of the bread samples. The formation of the trihydroxy fatty acid bitter compound, pinellic acid (9S,12S,13S-trihydroxy-10E-octadecenoic acid), was impacted by the lipoxygenase activity of the flour; however, there was no influence on the formation of 12,13-dihydroxy-9Z-octadecenoic acid or 1-(9Z,12Z-octadecadienoyl)-sn-glycero-3-phosphocholine. Additionally, the concentrations of all bitter compounds were significantly higher in bread made from aged flour versus non-aged flour.

Highly efficient oxidation of plant oils to C18 trihydroxy fatty acids by Escherichia coli co-expressing lipoxygenase and epoxide hydrolase

C18 trihydroxy fatty acids (THFAs), plant oxylipins, are used as antifungal agents and vaccine adjuvants.

Impacts of Adjunct Incorporation on Flavor Stability Metrics at Early Stages of Beer Production

The fresh flavor of a beer brand is one of its most important quality parameters and should be retained as long as possible through shelf-life. Early stages of beer production can have significant impacts on beer flavor stability. The use of cereal adjuncts – either malted or unmalted – is now widespread practice in brewing. This laboratory mashing study investigated the impacts of incorporating common solid adjuncts (corn and rice) at increasing grist percentages (0-50%) on flavor stability indicators measured in wort. Sweet worts were analysed for metal ion levels (ICP-MS), thiol content (RP-HPLC), oxidative stability (EPR spectroscopy), staling aldehydes (HS-SPME-GC-MS), trihydroxy fatty acids (GC-FID), t-2-nonenal potential (GC-MS), color and thiobarbituric acid (TBI). Mashing conditions for each formulation were adjusted in order to achieve target FAN values. Unmalted rice or corn adjuncts significantly reduced the total metal ion content, color, TBI and staling aldehydes in sweet worts at 50% adjunct incorporation (p < 0.05), relative in each case to the respective all-malt control mash. Our findings showed that unmalted rice or corn adjunct incorporation generally improved flavor stability metrics in sweet worts when similar mashing conditions were applied, whilst increasing the length of the ‘protein stand’ in mashing to reach target FAN in general had a negative impact on flavor stability indicators – e.g., wort T150 values significantly decreased in each recipe at 25% and 35% adjunct relative to the 12.5% adjunct brew but increased again at 50% adjunct, presumably due to the prolonged proteolytic stand employed to achieve target FAN content.

Identification of non-volatile compounds that negatively impact whole wheat bread flavor liking

Untargeted LC–MS flavoromic analysis was utilized to identify chemical compounds that impact consumer liking of whole wheat bread. Chemical fingerprints of thirteen whole wheat breads were modeled against consumer flavor liking scores by orthogonal partial least squares (OPLS) with good fit (R2Y = 0.98) and predictive ability (Q2 = 0.95). The four most predictive features (negatively correlated) were identified as 9S,12S,13S-trihydroxy-octadec-10E-enoic acid (pinellic acid), 9S,12S,13S-trihydroxy-octadeca-10E,15Z-dienoic acid, 8R*,9R*,10S*-trihydroxy-octadec-6Z-enoic acid, and 1-(octadeca-9Z,12Z-dienoyl)-sn-glycero-3-phosphocholine. Sensory validation studies including bitter threshold determination and recombination tests confirmed the contribution of these compounds to the perceived bitterness intensity of the bread samples and the overall negative impact on flavor liking. Lipoxygenase activity of the flour was reported to have a significant impact on the formation of the three bitter compounds (trihydroxy fatty acids) in the bread samples.

Microbial Synthesis of Linoleate 9 S-Lipoxygenase Derived Plant C18 Oxylipins from C18 Polyunsaturated Fatty Acids.

Plant oxylipins, including hydroxy fatty acids, epoxy hydroxy fatty acids, and trihydroxy fatty acids, which are biosynthesized from C18 polyunsaturated fatty acids (PUFAs), are involved in pathogen-specific defense mechanisms against fungal infections. However, their quantitative biotransformation by plant enzymes has not been reported. A few bacteria produce C18 trihydroxy fatty acids, but the enzymes and pathways related to the biosynthesis of plant oxylipins in bacteria have not been reported. In this study, we first report the biotransformation of C18 PUFAs into plant C18 oxylipins by expressing linoleate 9 S-lipoxygenase with and without epoxide hydrolase from the proteobacterium Myxococcus xanthus in recombinant Escherichia coli. Among the nine types of plant oxylipins, 12,13-epoxy-14-hydroxy- cis, cis-9,15-octadecadienoic acid was identified as a new compound by NMR analysis, and 9,10,11-hydroxy- cis, cis-6,12-octadecadienoic acid and 12,13,14-trihydroxy- cis, cis-9,15-octadecadienoic were suggested as new compounds by LC-MS/MS analysis. This study shows that bioactive plant oxylipins can be produced by microbial enzymes.

DIETHANOLAMIDE DERIVATIVES AS A POTENTIAL ENHANCED OIL RECOVERY AGENT FROM INDONESIAN CASTOR OIL AND USED FRYING OIL: ISOLATION, SYNTHESIS, AND EVALUATION AS NONIONIC BIOSURFACTANTS

Mono-, di- and trihydroxy fatty acids were reacted in a free-solvent amidation reaction with diethanolamine to obtain the derivative of dialkanolamides.The ricinoleic acid and 9,10,12-trihydroxystearic acid were isolated and prepared from Indonesian castor oil, while 9,10-dihydroxystearic acid was synthesized from the used frying oil. Amidation reaction of methyl ricinoleate, 9,10-dihydroxystearic acid and 9,10,12-trihydroxystearic acid gave the dialkanolamide derivatives in 93.10, 49.66, and 38.78% yield, respectively. The chemical structure of the synthesized products was elucidated by FTIR and GC-MS through silylation method. The prepared compounds were evaluated for their physicochemical properties as nonionic biosurfactant such as Hydrophilic-Lyophilic Balance value, interfacial tension, critical micelle concentration, foam stability, emulsification index, and emulsion stability. It was found that more hydroxyl groups contribute to a higher Hydrophilic-Lyophilic Balance value, foam stability, emulsification index, and lower interfacial tension.

Root herbivore performance suppressed when feeding on a jasmonate‐induced pasture grass

1. Plants defend themselves from insect herbivore attack using a range of physical and chemical defences which are in many cases regulated by phytohormones such as jasmonates. While much more is known about how jasmonates regulate defence against above‐ground herbivores (e.g. herbivores of leaves), there is increasing interest in how they influence below‐ground defences.2. For the Poaceae, most below‐ground studies focus on highly domesticated cereals. Here it is demonstrated how exogenous application of methyl jasmonate (MeJA) to the leaf blades of a non‐domesticated pasture grass (Microlaena stipoides) caused a more than two‐fold decrease in relative growth rate (RGR) of a root‐feeding chafer (Dermolepida albohirtum). MeJA treatment did not affect root consumption rates, but substantially reduced the efficiency of conversion of ingested food to body mass.3. Non‐targeted metabolomics identified significant changes in the metabolome of MeJA‐induced plants, with three compounds (a galactolipid, a trihydroxy fatty acid and a lysophospholipid) found to be correlated with herbivore RGR, although their roles in herbivore defence remain uncertain.4. This study suggests that an important Australian pasture grass can become better defended against root herbivores via enhanced jasmonate activity.

Identification of the molecular species of acylglycerols containing hydroxy fatty acids in wild edible mushroom Ganoderma lucidum

Abstract Edible Philippine mushrooms including Ganoderma lucidum have many health benefits. Seventy-two molecular species of triacylglycerols and five molecular species of diacylglycerols containing hydroxy fatty acids (FA) in the lipid extract of this mushroom were identified by HPLC and MS. The mono-, di- and tri-hydroxy FA constituents of the molecular species of acylglycerols were identified by various degrees of dehydration of [FA + Li] + . Many odd carbon numbered hydroxy FA constituents of acylglycerols were also identified. The contents of the 77 molecular species of acylglyerols and their constituent hydroxy FA in the lipid extract were estimated by the relative ion signal intensities of the molecular species of acylglycerols and the HPLC peak area % (ELSD). The contents (%) of the molecular species of acylglycerols containing hydroxy FA in the lipid extract were as: P-19:0-2OH18:2, 0.69%; 2OH18:2-2OH18:2-2OH18:2, 0.60%; P-OH19:1-OH19:1, 0.48%; S-OH19:1-OH19:1, 0.43%; 19:0-19:0-2OH18:2, 0.27% and the total of about 5.8%. “2OH18:2” stands for dihydroxy FA with 18 carbon atoms and two double bonds. The mushroom lipid extract was 1.67% of freeze-dried mycelium. The contents (%) of individual hydroxy FA in the mushroom lipid extract in decreasing order were: 2OH18:2, 1.4%; OH19:1, 1.1%; OH18:1, 0.38%; 2OH19:0, 0.19%; 2OH18:0, 0.13% and the total of about 3.5%. The odd carbon numbered hydroxy FA was about 1.44% of the mushroom lipid extract. The high contents of acylglycerols containing hydroxy FA, odd carbon numbered FA and odd carbon numbered hydroxy FA in edible mushroom might be related to one or more of its health benefits. As far as we are aware of, this mushroom contains the highest amounts of hydroxy FA and odd carbon numbered FA among the human foods.

Advances in Our Understanding of Oxylipins Derived from Dietary PUFAs.

Oxylipins formed from polyunsaturated fatty acids (PUFAs) are the main mediators of PUFA effects in the body. They are formed via cyclooxygenase, lipoxygenase, and cytochrome P450 pathways, resulting in the formation of prostaglandins, thromboxanes, mono-, di-, and tri-hydroxy fatty acids (FAs), epoxy FAs, lipoxins, eoxins, hepoxilins, resolvins, protectins (also called neuroprotectins in the brain), and maresins. In addition to the well-known eicosanoids derived from arachidonic acid, recent developments in lipidomic methodologies have raised awareness of and interest in the large number of oxylipins formed from other PUFAs, including those from the essential FAs and the longer-chain n-3 (ω-3) PUFAs. Oxylipins have essential roles in normal physiology and function, but can also have detrimental effects. Compared with the oxylipins derived from n-3 PUFAs, oxylipins from n-6 PUFAs generally have greater activity and more inflammatory, vasoconstrictory, and proliferative effects, although there are notable exceptions. Because PUFA composition does not necessarily reflect oxylipin composition, comprehensive analysis of the oxylipin profile is necessary to understand the overall physiologic effects of PUFAs mediated through their oxylipins. These analyses should include oxylipins derived from linoleic and α-linolenic acids, because these largely unexplored bioactive oxylipins constitute more than one-half of oxylipins present in tissues. Because collated information on oxylipins formed from different PUFAs is currently unavailable, this review provides a detailed compilation of the main oxylipins formed from PUFAs and describes their functions. Much remains to be elucidated in this emerging field, including the discovery of more oxylipins, and the understanding of the differing biological potencies, kinetics, and isomer-specific activities of these novel PUFA metabolites.

Spatiotemporal changes in the Content and Metabolism of 9, 12, 13 – Trihydorxy-10(E)-Octadecenoic Acid in Tomato (&lt;i&gt;Solanum Lycopersicum&lt;/i&gt; L. CV Balkonsar) fruits

In a previous study, we showed that 9,12,13-trihydroxy-10(E)-octadecenoic acid which is probably involved in plant defense responses is synthesized in tomato fruits and subjected to metabolism. Its catabolism or conversion was thus further characterized. The endogenous level of 9,12,13-trihydroxy-10(E)-octadecenoic acid decreased in the mesocarp and endocarp during the maturation of tomato fruits. The level of this trihydroxy fatty acid in the epicarp declined till the mature-green stage and then increased during the colour change. Injection of buffered solutions of unlabeled 9,12,13-trihydroxy-10(E)-octadecenoic acid into two weeks old fruits while they were still attached to the plants led to the alteration of contents of ten metabolites. After the infiltration of buffered solutions of isotopically labeled 9,12,13-trihydroxy-10(E)-octadecenoic acid into green unripe tomato fruits which were still attached to the plant, the highest radioactivity was found in compounds much more polar than 9,12,13-trihydroxy-10(E)-octadecenoic acid and whose signals in HPLC-UV-chromatogram coincided with the citric acid peak. These results indicated that 9,12,13-trihydroxy-10(E)-octadecenoic acid could be implicated in the control of tomato fruit maturation and β-oxidation might be one of its degradation pathways Keywords : Ethylene production, fruit maturation, ripening, β-oxidation, pinellic acid, Solanum lycopersicum L. cv Balkonstar

Identification of TAG and DAG and their FA Constituents in Lesquerella (Physaria fendleri) Oil by HPLC and MS

AbstractCastor oil has many industrial uses because of its high content (90 %) of the hydroxy fatty acid, ricinoleic acid (OH1218:19). Lesquerella oil containing lesquerolic acid (Ls, OH1420:111) is potentially useful in industry. Ten molecular species of diacylglycerols and 74 molecular species of triacylglycerols in lesquerella (Physaria fendleri) oil were identified by electrospray ionization mass spectrometry as lithium adducts of acylglycerols in the HPLC fractions of lesquerella oil. Among them were: LsLsO, LsLsLn, LsLsL, LsLn–OH20:2, LsO–OH20:2 and LsL–OH20:2. The structures of the four new hydroxy fatty acid constituents of acylglycerols were proposed by the MS of the lithium adducts of fatty acids as (comparing to those in castor oil): OH1218:29,14 (OH1218:29,13 in castor oil), OH1218:39,14,16 (OH18:3 not detected in castor oil), diOH12,1318:29,14 (diOH11,1218:29,13 in castor oil) and diOH13,1420:111 (diOH20:1 not detected in castor oil, diOH11,1218:19 in castor oil). Trihydroxy fatty acids were not detected in lesquerella oil. The differences in the structures of these C18 hydroxy fatty acids between lesquerella and castor oils indicated that the polyhydroxy fatty acids were biosynthesized and were not the result of autoxidation products.

Inhibition of Prostaglandin D2 Production by Trihydroxy Fatty Acids Isolated from Ulmus davidiana var. japonica

The stem and root barks of Ulmus davidiana var. japonica (Ulmaceae) have been used to treat inflammatory diseases including mastitis, rhinitis, sinusitis, and enteritis. In an ongoing study focused on the discovery of natural anti-inflammatory compounds from natural products, a methanol extract of the stem and root barks of U. davidiana var. japonica showed anti-inflammatory activities. Activity-guided fractionation of the methanol extract yielded a new trihydroxy fatty acid, 9,12,13-trihydroxyoctadeca-10(Z),15(Z)-dienoic acid (1), and a known compound, pinellic acid (2). These two trihydroxy fatty acids 1 and 2 inhibited prostaglandin D₂ production with IC₅₀ values of 25.8 and 40.8 μM, respectively. These results suggest that 9,12,13-trihydroxyoctadeca-10(Z),15(Z)-dienoic acid (1) and pinellic acid (2) are among the anti-inflammatory principles in this medicinal plant.

Overexpression of hydroperoxide lyase, peroxygenase and epoxide hydrolase in tobacco for the biotechnological production of flavours and polymer precursors

Plants produce short-chain aldehydes and hydroxy fatty acids, which are important industrial materials, through the lipoxygenase pathway. Based on the information that lipoxygenase activity is up-regulated in tobacco leaves upon infection with tobacco mosaic virus (TMV), we introduced a melon hydroperoxide lyase (CmHPL) gene, a tomato peroxygenase (SlPXG) gene and a potato epoxide hydrolase (StEH) into tobacco leaves using a TMV-based viral vector system to afford aldehyde and hydroxy fatty acid production. Ten days after infiltration, tobacco leaves infiltrated with CmHPL displayed high enzyme activities of 9-LOX and 9-HPL, which could efficiently transform linoleic acid into C(9) aldehydes. Protein extracts prepared from 1 g of CmHPL-infiltrated tobacco leaves (fresh weight) in combination with protein extracts prepared from 1 g of control vector-infiltrated tobacco leaves (as an additional 9-LOX source) produced 758 ± 75 μg total C(9) aldehydes in 30 min. The yield of C(9) aldehydes from linoleic acid was 60%. Besides, leaves infiltrated with SlPXG and StEH showed considerable enzyme activities of 9-LOX/PXG and 9-LOX/EH, respectively, enabling the production of 9,12,13-trihydroxy-10(E)-octadecenoic acid from linoleic acid. Protein extracts prepared from 1 g of SlPXG-infiltrated tobacco leaves (fresh weight) in combination with protein extracts prepared from 1 g of StEH-infiltrated tobacco leaves produced 1738 ± 27 μg total 9,12,13-trihydroxy-10(E)-octadecenoic acid isomers in 30 min. The yield of trihydroxyoctadecenoic acids from linoleic acid was 58%. C(9) aldehydes and trihydroxy fatty acids could likely be produced on a larger scale using this expression system with many advantages including easy handling, time-saving and low production cost.

Ratios of Regioisomers of Minor Acylglycerols Less Polar than Triricinolein in Castor Oil Estimated by Mass Spectrometry

AbstractWe have recently reported the identification of forty new minor molecular species of acylglycerols containing hydroxy fatty acids less polar than triricinolein by electrospray ionization mass spectrometry of the lithium adducts. The ratios of regioisomers of triacylglycerols (ABC and AAB types) and tetraacylglycerols (AAAB type) identified were estimated by the relative abundances of the fragment ions from the neutral losses of fatty acids as α,β‐unsaturated fatty acids at the sn‐2 position. The order of the contents of regioisomers of triacylglycerols with the fatty acids at the sn‐2 position are: nonhydroxy fatty acids &gt; monohydroxy fatty acids &gt; dihydroxy fatty acids &gt; trihydroxy fatty acids. For tetraacylglycerols (AAAB type) such as ricinoleoylricinoleoyl–ricinoleoyl–oleoyl–glycerol (RRRO), ricinoleoylricinoleoyl chain was predominately at the sn‐2 position, while ricinoleate was not detected at the sn‐2 position.

Production of a Novel 9,12-Dihydroxy-10(E)-eicosenoic Acid from Eicosenoic Acid by Pseudomonas aeruginosa PR3

Microbial conversions of unsaturated fatty acids often generate polyhydroxy fatty acids, giving them new properties such as higher viscosity and reactivity. A bacterial strain Pseudomonas aeruginosa (PR3) has been intensively studied to produce mono-, di-, and trihydroxy fatty acids from different 9-cis-monoenoic fatty acids such as oleic acid, ricinoleic acid, and palmitoleic acid. However, from the results and the postulated similar metabolic pathways involved in these transformations, it was assumed that the enzyme system involved in transformation of the monoenoic fatty acid by strain PR3 could utilize fatty acids with different chain lengths and locations of the double bond. In this study was used as a substrate for bioconversion by strain PR3 eicosenoic acid (C20:1, ω-9) containing a singular cis double bond at different positions from the carboxyl end as oleic acid, and it was confirmed that PR3 could produce a novel 9,12-dihydroxy-10(E)-eicosenoic acid (DED) with 6.2% yield from eicosenoic acid. The structure of DED was confirmed using GC-MS, FTIR, and NMR analyses. DED production was maximized at 72 h after the substrate was added to the 24 h culture. Some other nutritional factors were also studied for optimal production of DED.

A new trihydroxy fatty acid ester from the root of Isatis indigotica

From the EtOAC extract of the root of Isatis indigotica, a new trihydroxylated monounsaturated fatty acid ester 1, together with two known fatty acids 2–3 that were separated for the first time, has been isolated. The structure of this new linoleic acid-derived metabolite was established as methyl-8,11,12-trihydroxy-9Zoctadecenoic acid on the basis of spectroscopic data.

Thermostable lipoxygenase is a key enzyme in the conversion of linoleic acid to trihydroxy-octadecenoic acid by Pseudomonas aeruginosa PR3

Lipoxygenases (LOXs) constitute a family of lipid-peroxidizing enzymes that catalyze the oxidation of unsaturated fatty acid containing a (1Z,4Z)-pentadiene structural unit, leading to formation of conjugated (Z,E)-hydroperoxydienoic acid. LOXs are known to be widely distributed in plants and animals. Recently, several microbial LOXs were reported to be involved in the production of hydroperoxy fatty acids. Among the microorganisms that produce hydroxy fatty acids, Pseudomonas aeruginosa PR3 is known to convert linoleic acid to trihydroxy fatty acid, which suggests the involvement of a LOX enzyme. Based on these reports, we identified a novel thermostable LOX from P. aeruginosa PR3 strain. The protein was purified 34.3-fold with a recovery rate of 5.14%. The Km and Vmax values of the purified enzyme were 3.57 mM and 0.73 μmol/min//mg, respectively. Heat stability of the purified enzyme was unexpectedly high with an LD50 of 90 min at 80°C, although P. aeruginosa PR3 is known as a mesophilic bacterium. Substrate specificity of the purified enzyme was restricted only to unsaturated fatty acids carrying a (1Z,4Z)-pentadiene unit.