1294

lions. The kinetic studies showed that the esterase and thicesterase activi-ties of the His-lagged enzyme were similar to those of the wild type, Onthe other hand. the catalytic efficiency of chymouypsin-like activity of theHis-tagged protein was two times higher than that of the wild type.JAOeS 74, J37H376 (1997).

ABSTRACTS FROM AOCS JOURNALS

Biocalalyllc Synthesis of Some Chiral Drug Intermediates by Oxi-doreductllSe$. Rarnesh N. Patel·, Ronald L. Hanson, Amil Baneljee. andLaszlo J. Szarka. Departmem of Microbial Technology, Bristol-MyersSquibb Pharmaceutical Research Institute. New Brunswick. New Jersey08903.

Chiral intermediates were prepared by biccatalytic processes with oxi-doreductases for Ihe chemical synthesis of some pharmaceutical drug can-dictates. These include: (i) the microbial reduction of [-(4-fluorophenyl}4-[4-(5-f1uoro-2-pyrimidinyl} l-piperuinyl]-I-tJluanolle (I) to R-( +)-1-(4-f1uorophenyl)-4-14-(5-f1uoro-2-pyrimidinyl)-I-piperazinyl]-l-butanoI (1)[R-{+)-BMY 14&02], an antipsychotic agent; (ii) the reduction of N-4-(I-oxo-z-chtoroaceryl ethyl) phenyl methane sulfonamide (3) to the corre-spending chiral alcohol (4). an intermediate for 0-(+)-N4-{I-hydroxy-2-[(-methylethyl)aminolethyl}phenyl methanesulfonamide [0-(+) soralol], aIJ-blocker with class III antiarrhythmic properties; (iii) biotransformationof NE-curbobenzoxy (CBZ)-1.-lysine (7) to NE-CBZ-1.-oxylysine (5). anintermediate needed for synthesis of (S)-J.[6-umino-2-([hydroxy{4-phenyl butyl)phosphinyl]oxy) l-oxohexyll-t-proline (ceronapril). a newangiotensin convening enzyme inhibitor (6) and (iv) enzymatic synthesisof L-j3-hydroxyvaline (9) from u-keto-jl-bydroxyisovalerate (16). 1.-13-Hydroxyvaline (9) is a key chiral intermediate nceded for the synthesis ofS-(Z)-{{I-(2-amin0-4-thiazolyl)- 2- ( [2.2-<1imethyI4-oxo-l-(sulfooxy)- 3-azetidinyl] aminoJ-2-oxocthylidene]aminoloxyacetic acid (tigemonam)(10). an orally active monobactam.JAOCS 74. 134S-1360{l997).

Chemoemymatjc Approaches to SCH 56592, A New Azole Antifun-gal. Brian Morgana.*. Brent R. StockweW". David R. Doddss. David R.Andrewsb• Anantha R. Sudhakarb, Christopher M. Nielsenb. IngridMergelsbergv, and Arne Zumbachv. aSchering-Plough Research Institute,Bio\Tansformations Group, K-15-1I1800. Kenilworth. New Jersey 07033-0539. bChemical Process R&D, Union. New Jersey, 07083, and"Wenhenstein Cbemie AG, CH-6i05 Schachen. Switzerland.

Chemocnzymatic approaches 10 the synthesis of two tey chiral pre-cursors of a new uole antifungal agent. SCH 56592, are described. Inparticular. the enzymatic dasrereosetecnve acylation of 2-benzyloxy-3-peruanol (7) was developed to produce (2S.3R)-7 in >97% diastereomericexcess (de) from otherwise unusable mixtures of (2S.3R)/(2S.3S)-7(40-80% de). The selectivity and reactivity of commercially availableCandido "'sosa and Mucor miehei liposes are compared for the acylationof 7 and the hydrolysis of the corresponding butyrate 16a. Of the 17 Crugosa enzyme preparations that were examined for acylation of 7. twopurified enzyme preparations showed no reactivity, five enzymes showedhigh diastereoselectivity with preference for the (2S.3R)-isomer. andseven showed a slight preference for the (2S.3Sj-isomer.JAOCS 74. 1361-1370 (1997).

Facile Puriftcatioo of a C-Terminal Extended His- Tagged Vibrio mim-icus Arylesterase and Characterization or the Purified Enzyme. Ya-Lin Lee". Rey-Chang Changb. and Jei-Fu Shaw"*, alnstitute of Botany.Academia Sinica. Nankang. Taipei. Taiwan 11529. and bDepanment ofSea-Food Technology. China College of Murine Technology, Taipei. Tai-wan Ill.

Vibrio mimicus arylesterase. a 20 kDa protein. is a multifunctional en-zyme with rntoesierase and chymotrypsin-like activities. Because an affin-ity His-tag (six consecutive histidine affinity tag) directly to the proteincaused the loss of enzyme activity_ a bexadecapeptide with His-tag, ADP-NSSSVDKLAAALEHHHHHH encoded from vector pET-20b(+) wasconstructed to extend from the carboxyl terminus of the erylesrerese. ThisHis-tagged protein retained enzyme functions. Thermal unfolding behaviorof both pr(ltcins was almost identical. and their Tm values were near 54"Cas monitored by circular dichroism. Tryptic cleavage of the functional His-tagged enzyme produced tWOsmaller proteins. which still possessed en-zyme activity and which suggested that the ad<.lilional peptide extended onthe pr(ltein surface. The spacing peptide between His-tag and arylesterasesuccessfully prevented the interference of the His-tag to the enzyme func-

Purincalion and Characterization or a Cotd-Acuve Protease rromPsychrotrophic Serreuia marcescens AP3801. Yasutaka Moritua, KenjiKondoh'", Quamrul Hasanb. Toshifumi Sakaguchiv. Yuji Murakami".Kenji Yokoyamav. and Eiichi Tamiya(l·*, "School of Materials Science.Japan Advanced Institute of Science and Technology. Tutsunckuchi,Ishikawa 923-12. Japan. aod bResearch and Development Division, Ana-lytical. Technology and Research Depanmenl, Procter & Gamble FarEast, Inc., Higashinada-ku. Kobe 658.lapan.

Protease activity was detected in the culture medium of Serratiamarcescens AP380i grown atW"C, which was isolated from soil collectedfrom the top of a mountain. The enzyme. designated as CP-58 protease,was purified to homogeneity from the culture broth by ion exchange andgel filtration chromatographies. The molecular mass of the protease was58 kDa. and its isoelectnc point was close to 6.0. Maximal activity towardezocesein was observed at 40'C and from pH 6.5 to 8.0. The activity wasstrongly inhibited by I,IO-phennnthroline. suggesting that the enzyme is ametalloprotease. The N-terminal amino acid sequence was Ser-Leu-Asn-Gly-Lys- Thr-Asn-Gly- Trp-Asp-Ser- Val-Am- Asp-Leu-Leu-Asn-Tyr-H isAsn-Arg-Gly-Asn (or Asp)-Gl y-Thr-Ile-Asn-Asn-Lys- Pro-Ser-Phe-Asp-lie-Ala. A search through databases for sequence homology aligned CP-58 protease with meLalioprotease. llIe result of the cleavage pallem of oxi-dized insulin B-chain suggests that CP-58 protease has a broader speci-ficity than other proeases against the peptide substrate.JAOCS 74. 1377-1383 (1991).

znenuosereeuve Formalion of an u, IJ·Epoxy Alcohol by Reaction ofMethyl 13(S)-Hydroperoxy-9(Z),II(E)-octadecadienoate with Titanl-um Isoprcpcxlde. George J. Piazza". Thomas A. Foglia. and AlbertoNuhez , ERRC, ARS. USDA. Wyndmoor, Pennsylvania 19038.

Methyl II(R).12(R)-epoxy-13{S)-hydroxy-9(Z)-octadecenoate (threQisomer) was generated from linoleic acid by the sequential action of anenzyme arKIrwo chemical reagents. Linoleic acid was treated with llpoxy-genase to yield its corresponding hydroperoxide [13(S)-hydroperollY-9(Z).II{E)-octadecadienoic acid]. After methylation with CH2N2. thehydroperoxlde was treated with titanium (IV) lsopropcxide [li(O-i-Pr)4]at 5°C for 1 h. The products were separated by normal-phase high-perfor-mance liquid chromatography and characterized with gas chromatogra-phy-mass spectrometry, infrared spectroscopy, and nuclear magnetic reso-nance spectroscopy. Approximately 30% of the product was methyl 13(S)-hydroxy-9(Z),11(E)-octadecadienoate. Over 60% of the isolated productWIl5 methyl II(R).12(R)-epoxy-13(S)-hydroxy-9(Z)-octadecenoatc. Aftcrquenching li(O-i-Pr)4 with water, the spent catalyst could be removedfrom the fatty products by partitioning between CH2Ci2 and water. Theseresults demonstrate that li(O-i-Pr)4 selectively promotes the formation ofan c-epoxtde with the thrro configuration. It was critically important tostan with dry methyl 13(S)-hydroperoxy-9(Z),II(E)-octadecadienoatebecause the presence of small amounts of water in the reaction mediumresulted in the complete hydrolysis of epoxy alcohol to trihydroxy prod-ucts.JAOCS74,1385-1390(1997).

Characterization of New Yeast Llpuses. Ching T. Hou". Oil ChemicalResearch. NCAUR, ARS, USDA. Peoria. Illinois 61604.

Previously, we used a simple. sensitive agar plate method to screenlipase activity from 1229 selected cultures. including 508 bacteria, 479yeasts. 230 actinomycetes and 12 fungi. that covered many genera andspecies. About 25% of the cultures tested were lipase-positive. We alsoexpanded our screening methOtl to focus specifically on the pH depee-dence and tbermostabilhy of these lipase activities. In this report. we havecharacterized 25 yeast Iipeses. obtained from our screening program, onthe basis of their positional specificity against uiglycerides. Lipase wasproduced by growing cultures on nutrient medium in the presence of ves-etable oil at 25°C for 4 d. Of the 25 new yeast llpases analyzed, 19showed 1.3-positional specificity Hnd 6 showed random specificity. No 2-positional specific bpases were found. Among those cultures with highestlipase activity are: Candida sil ..icola NRRL YB-2846 (random): Cimdid(l

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sp. 55 (random); Clint/ida sp. 125 (random): Pichio Q~rica1Ul NRRL y-21S6 (I.3·specilic): P. muscic% NRRL Y·1()(J:5 (random); P. peremlniiNRRL YB-3808 (l.3-specific); and YarmlO'ja IipolytiCQ NRRL YB-423(random). CharacteriUltion or Canditk! sp. strain 55 lipase on ns substratepreference showed Ihllt (hi) enzyme llydTOlyud soybean oil triglyceridespecies LLLn. LlL. LLO, and LLP TnOrI: readily than roo. LOP. 000.LOS, and roo, where L • linoleic, Ln :0 linolenic, 0 '" oleic, P ..palmitic. and 5 _ stearin.lAOCS 74,1391-1394 (1997).

Changes in Hydrolysis Specificities or Lipase rrom Rhi:omucormidei 10 PoI)"unsatunlttd FaUy Ac)"1 Eth)"] Esters in Different Aggre-piton Slalrs. YmhilSUgu Kosugia.·. Qing-Iong ClIangb. Kenji Kanau-wao, and Hiroshi Nakanishi':!. °N.liona] Institute of Bioscience andHuman-Technology, T5ulrJ.a~ 305, Japan. and b:tnstitutc of Cbemical Met-allurgy_ Chinese Academy of Science. Beijing. P.R. China.

Hydrolysis specificities of lipase from Rhizomucor miCMi were com-pared for various fany acyl ethyl esters. Activity yields of immobili:ted Ii-pases, me:asu~ with I mM subslnte. were more than 1()()Il,. Differencesin hydrolysis rate and amnity for the subsuares between lipase prepara-tions were also typically higl1cr durinll hydrolysis of substratcs at 100 mMthan at I mM, indk:ating better mass transfer effects for l-mM substrates.The IIDlive lipase showed higher affinity for polyun~tunted fany acid sub-strates all mM than at 100 mM. Hydrolysis eaes for l·mM substrateswere observed with immcbitized llpases. fixed on anion exchange resinwith glutaraldehyde and on cation exchange carrier with carbodiimide. andsUlLgested some: modification of the bask: amino acid related to the lid ofR. miehei lipase. Activation with these bifunctional reagerns was not 0b-served for 1000mM SUbstrates, indicltinll !hat interfacial activation alwaysoccurred becaU5Cof aggrellalion of lOO-mM substrates. These resuns showthat lipase from R. miehei ~nizes molecular aggregation of lipids. andthat various changes occur in the hydrolysis specificities for fally acids.JAOCS 74. J39S-1399 (1991)

Additive Effects or Aeyl·Blndlng Site Mulations on the Fatly AddSeleclh'lty or Rhi~opus dc1emor Lipase. Roben R. Klein". GregoryKing, Rebert A. Moreau, Gerald P. McNeill. Pierre Villeneuve, andMichael J. Haas, ERRC, ARS, USDA, Wyndmoor, Pennsylvania [9038.

The falty acid specificity and pH dependence of triacylglycerol hydrol-ysis by the RhillJPus dc/cmar lipase acyl·binding site mutant Val206Thr +PIle95Asp (Val, valine; Thr, threonine; PhI', phenylalanine; Asp, asparticacid) were characterized, The activity of the double mutant prolipase wasreduced by as much as IO-fold. compared to the wild-type prolipase. How-ever, the fatty acid specificity profile of the enzyme was markedly sharp-ened and was dependent on the pH of the substrate emulsion. At neutralpH. strong preference (IO-fold or greater) for hydrolysis oftriacylglycerolsof mediurn-ebainlength fany adds (C8:0 to CI4:0) was displayed by thevariant prolipase. with no hydrolysis ortrtecylglycerols of shon-chain fanyacids (C4:0 to C6:0) and little activity manifested towlltd fauy acids with16 or more carbons. At acidic pH values. the fauy acid setectiviry profileof the double mutant prolipa!>e expanded to include short-chain triacyl-glycerols (C4:00 C6:0)' When assayed against a triacylglycerol mi:lture oftribulyrin. tricaprytin and triolein. the Va1206Thr + Phc95Asp prolipasedisplayed a high selectivity for caprylic acid and released this fauy acid atIC8.I't2j·fold more efficiently than the others present in the substrate mix-ture. When presented a mixture of nint Iany acid methyl esters. the wild-type proupase sho .....ed a broad Substlllte specificity profile. hydroly~ingthe various methyl CStcn to a ~imililf extent. Contrastingly. the double mu-tanl prolipase displayed a narro .....ed substrate specificity profile. hydrolyz-ing caprylic methyl cster at nearly wild·type levels, while its activityagainst the other me:thyl esters examined was 25· 10 5·fold lo.....er than !hatobserved for the wild-lYpe enzyme.JAOCS 74, 1401-1401 (1991).

Enzymatic Synthesis of P05ltlon·Sp«tnc Lo .....-Calorie StructuredLipids. Casimir C. Akoo* ~nd Lisa N. Yee. Depanmcnt of Food Scienceand Technology. Food Science Building. The Uni"ersity of Georgia,Athens. Geofxi. 30602.

An immobili:ted sn-I,3'5pecific lipase from Rhi:omuCQr mithei (1M60) was used 10catalyze the inteI"CStcriflCuion of tristearin (C 18:0> and tri-caprin (CIO:O) to produce low-<:alone structured lipids (SL). AcceptablepnxIuct yields were obtained from. 1:1 mole ntio ofboth triacylglyccrols

with I~ (wlw of reactants) of 1M 60 in 3 mL hexane. Tbe SL molecularspecies. based on total carbon number. were 44.2'1. C41 and 40.5'1. C49'with 3.8 and I 1.5'lounreacted tristearin C51 and tricaprin C21. respec·tively. remaining in the product mixture. The best yield of C41 species(44.3"') was obtained with uro added water. Tricaprylin (C8:0) was alsosua:essfully intereslerified with tri5learin in good yields at 1:1 mole ratio.ProdOCIS were analyzed by reverse-phase high- performance liquid chro-matography with an evaporative light·scattering detector. Reaction peram-erers, weh as substraie mole rauo. enzyme: load. time: course. added water,reaction media, and enzyme reuse, were also investigated, Hydrolysis bypancreatic lipase revealed the specific falty acids present at the slI·I.3 po-sitions of SLJAOCS74,14C»-1413(1991).

Water Acth·\ty-AdJusltd Enzymatic hr1ial IIfdrolysis of Phospho-lipids to Concentrate Pol"ullSDluraled Fauy Acids. MaSlyo Ono".Masashi Hosoklwlo. Yoshiklzu Inoueb. and Koretaro Takahashio,-,lfJ...aboratcwy of Food Biochemistry Faculty of Fisheries. Hokkaido Uni-va1ity. Hakodate 041. Japan. and bBiun Chemical Co .. Lrd.. Okayama109·m, Japan.

Selective panial hydroly!loes of egg yolk phospholipid and squid skinphospholipid were carried OUt. By kcc:ping the water aClivily (a",) ofLipozyme: 1M at an intermediate level. it was easy to concentrate eccose-hexacnok: acid (DHA). It was also possible to concentrate both DHA andarachidonic acid (AA) simultaneously to a cenain level under this a",range. However. it was impossible to concentrate AA alone when DHAwas present. Though there is a limitation in concentrating AA exclusi~ly.the proposed aw-adjusted hydrolytic reaction is a promising way rorpreparing phospholipids rich in DHA.JltOCS 74, 141S-1411 (1997).

The Prepanllon or Coneenlrates or Elcosapentaenoic Acid andDocosahexatnoic Acid by Lipasc--Catalyud TrMnSC'Sterificallon orFish Oil wlth Ethanol. Gudmundur G. Haraldssona.-, BjOrnKrhtinuona, Ragnheidur Si:\urdardottirO• Gudmundur G.Gudmuntissona. and Harald Breivik . aScience Institute. Uni\'ersityofIceland, Reykjavik, Iceland. and bNonk Hydro ASA. Research CentrePorsgrunn. Poesgrunn. Norway.

The objective of this study was to investigate the use of lipases as cata-lysts for producing concentrates of eiccsaperuaenoic acid (EPA) and do-ccsahexaenclc acid (DHA) from fish oil as an alternative to con"entionalchemical procedures. Transcsterification of fish oil with ethanol was con-ducted under anhydrous solvent-free conditions with a stoichiometricamount of ethanol. Among the 11lipa!>es tested. the results showed thatPseudomonas1ipll5tS had the highest activity toward the saturated and m0-

nounsaturated fauy acids in the fish oil. much lower activity toward EPAand DHA and. Pt the same time, good tolerance toward the anhydrous el-coholic conditions. With 10 wt'll of lipase. based on weight of the fish oiltriacylglycerol substrate (15'" EPA and 9 ... DliA initial content). a 50tJbconversion into ethyl esters was obtained in 24 h at 20"C. in which timethe bulk of the saturated and monounsaturated fatty acids reacted, leavingthe long-chain n-) polyunsaturated fatty acids unreacred in the residualmixture as mono-. di-. and triacylglycerols. This miJHure comprised ap-proximately.5O'l> EPA + DIiA. Total recovery of DHA and EPA was high,over 8a... for DHA and more than 9O'lo for EPA. The observed fany acidselectivity. favoring DHA as a subMrate. was most unusual because mostlipases favor EPA.JltOCS 74, 1419-1424 (1991).

Preparation or Highly Purined Concentrates or EirnsapentaenokAcid and Docosahuaenoie Acid. Harald Breiviko,-. Gudmundur G.Harnldssonb, and BjOrn Kristinssooh. uNorsk Hydro ASA. Research Cen-tre Porsgrunn, N-3901 Porsgrunn, Norway, and bscience Institute. Uni-venity of Iceland. Dunhaga 3. 15-101 Reykjavik. Iceland.

Because of the complexity of marine lipids. polyun~lUrated fany acid(PUFA) derivatives in hiibly purified form ere not easily prepared by anysingle fntCtionation technique. The products are usually prepared as theethyl esters by esterification of the body oil of fat fish species and subse-quent physicochemical purification processes. including sllon-path dislil·lettcn. urea fntClionation. and preparative chromatography. Lipase-cat-alyzed transoesteriflCation has been shown to be an excellent alternative 10tnlditional esterification and shan·path distillation for concentrating the

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ABSTRACTS FROM AOCS JOURNALS

combined PUFA-<:ontent in fish oils. At room temperature in the presenceof PU:uMmorUlS sp- lipase and a stoichiometric amount of ethaool withoutany solvent. efficient transesterification of fish oil was obtained. At 52%conversion, a concentrate of 46% eicosapentaencic acid (EPA) plus do-oosaheucnoic acid (DHA) was obtained in excellent recovery as a mi:ttureof mcoo-, di-, and triacylglycero\s. The latter can be easily separated fromthe saturated and monounsaturated ethyl esters and convened into ethylesters either by conventional chemical means or enzymatically by immo-bilized Candido. (J.fllarr:tic(J lipase. Urea-fractionation of such an interme-diary product can give an EPA + DHA cement of approximately 85%.JAOeS 74. 1425--1429 (1997),

Production or High Yields or Dccesabexeenoic Add by SchiZiJChytri·um sp. Strain SR21. T. Yaguchiu .• , S. Tanakab, T. Yokochic• T. Naka-harac, and T. Higashiharav. aSuntory Lid .. Shimamoto-cho. Osaka 618.Japan, bNagase Biochemicals Ltd., Fulmchiyama, Kyoto 620, Japan. andCNational Institute of Bioscience and Human-Technology, Tsukuba,lbaraki 305, Japan.

The culture conditions for high-yield production of docosahexaenoicacid (DHA) by SchizochYlrium sp. strain SR21 were investigated in a fer-menrer. With increasing carbon (glucose) and nitrogen (com steep liquorand ammonium sulfate) sources (up to 12% glucose) in the medium. DHAproductivity increa<;ed without a decrease in growth rate, i.e., 2.0. 2.7, and3.3 g DHAll.Jd with 6, 10, and 12% glucose, respectively. Eventually, 48.1g dry cells/L and 13.3 g DHAIL were produced in 4 d with 12% glucose.DHA productivity was decreased with 15% glucose, i.e., 3.1 g/Ud. With12% glucose. the lipid content was 77.5% of dry cells. and DHA contentwas 35.6% of total fatty acids. The lipid was composed of about 95% neu-tralIipid and 5% polar lipid. In polar lipids, the contents of phospbarldyl-choline (PC). phosphatidylethanolamine, and phcsphatldyllnoslto! were74. II. and 5%, respectively. The PC profile was simple, 70% nf PCmolecules were l-palmitoyl-2-DHA-PC and 1.2-di-DHA-PC. These re-sults indicate that Sch;ZQchy/rium sp. strain 21 is an excellent source formicrobial DHA production. including not only the acid form of DHA butalso 2-DHA-PC.JADeS 74, 1431-1434 (1997).

Industrial Uigh_Perlorman~ Liquid Chromatography Purification ofDecesahexaenclc Acid Ethyl Ester and Docosapentaenoic Acid EthylEster Irum Single-Ceil 011. R. Yamamura and Y. Shimomura·. YMCCo., Ltd .. 61-] Tal-Sbinarami, Kumiyama-Cho Kuse-Gun, Kyoto. 613Japan.

The use of polyunsaturated fatty acids (PUFA) as medicine or in func-tiona] diets requires high purity. An industrial purification method forPUFA from Schizochytrium sp. SR21 oil was investigated. This oil con-tains fewer unwanted components than fish oils. Docosahexaenoic acidand docosapentaenoic acid ethyl esters (DUA-E and DPA-E) were pre-pared by treatment of this oil with ethanol and I N potassium hydroxide inhexane. DHA-E and DPA-E were purified by an industrial high-perfor-mance liquid chromatography (UPLC) plant. The separation plant consistsof two columns (400 mm i.d., ].000 mmL) with temperature-controlledwater jackets and double-plunger (four heads) injection and eluent pumps.This plant was computer-controlled and equipped with an explosion-pre-vention system. The packed material was octadecylsilica (reverse-phaseODS), and the eluent was methyl a1cohoiJwater (98:2). DHA-E and DPA-E from single-cell oil were highly purified by this industrial HPLC methodin a one-step process. The DHA-E and DPA-E obtained were better than99% purity.JAOeS 74,1435--1440 (1997).

Purinution nr Docosahexaenoic Acid Irem Tuna Oil by a Two-StepEnzymatic Method: UydroltSis and Selective Esterification. Yu~ Shi-madaa,., Kuuaki Maruyama, Aldo Sugiharaa, Shigeru Moriyama, andYoshio Tominaga'", aOsaka Municipal Technical Research Institute.Osaka 536, Japan. and i>Maruha Corp., Ibaraki JOO..42. Japan.

Purification of docosahexaenolc acid (DHA) was allempted by a two-Step enzymatic method that consisted of hydrolysis of tuna oil and selectiveesterification of !he resulting free fany acids (FFA). When more than 60%of tuna oil was hydrolyzed with Pseudomonas sp. lipase (Lipase~AK), theDHA COntent in the FFA fraction coincided with its content in the originaltuna oil. This lipase showed stronger activity on the DHA ester than on theeicosapenlaenoic acid ester and was suitable for preparation of FFA rich in

DHA. When a mixture of 2.5 g tuna oil, 2.5 g water, and 500 units (U) ofLipase-AK per 1 g of the reaction mixture was stirred at 400c for 48 h,83% of DHA in tuna oil was recovered in the FFA fraction at 79% hydrol-ysis. These fatty acids were named tuna-FFA-Ps_ Selective esterificationwas then conducted at JOOc for 20 h by stirring a mixture of 4.0 g of tune-FFA-Ps/lauryl alcohol (1:2. moVmol), 1.0 g water. and 1,000 U of Rhiw-pus delemnr lipase. As a result. the DHA content in the unesteritled FFAfraction could be raised from 24 to 72 wt% in an 83% yield. To elevate theDHA content further, the FFA were extracted from the reaction mixturewith a-hexane and esterified again under the same conditions. The DHAcontent was raised to 91 wt% in 88% yield by the repeated esterification.Because selective esterification of fauy acids with lauryl alcohol proceededmost efficiently in a mixture that contained 20% water. simultaneous reac-tions during the esterification were analyzed qualitatively. The fauy acidlauryl esters (L-FA) generated by the esterification were not hydrolyzed.In addition. L-FA were acidolyzed with linoleic acid. but not with DHA.These results suggest tharlauryl DHA was generated only by esterification.JADeS 74,1441-]446 (]997).

Carica papaya Latex-Catalyzed Synthesis of Structured Trlacylglyc-erois. T.A. Foglia" and P. Villeneuve, USDA, ARS, ERRC. Wyndmoor,Pennsylvania 19038.

One impediment to the industrial use of enzymes in fat and oil trans-formations is the higher cost often associated with an enzymatic processcompared with the corresponding chemical process. Processes that utilizeplant enzymes, however. may have advantages because of their lower costand ready availability. One example of such a plant-derived enzyme isCarica papaya latex (CPL). the principal source of the protease papain.Recently, it has been shown that this latex also catalyzes the lipolysis oftriacylglycerols and that this latex lipase has a selectivity for short-chainacyl groups as well as a I ,3-glycerol selectivity. These selectivities can beused in the synthesis of structured rriacylglycerols. In this paper we de-scribe the utility of CPL in lipase-catalyzed reactions, specifically the syn-thesis of low-calorie triacylglycerol analogs.JADCS 74. 1447-1450 (1997).

bnprovement or Palm Oil Through Breedtngaed Biotechnology, B.S.Jalani'", S.c. Cheah. N. Rajanaidu, and A. Darns. Palm Oil Research insti-tute of Malaysia, 50720 Kuala Lumpur, Malaysia.

lbe oil palm Elaeis guineensis is the highest oil-yielding crop and hasthe potential to become the major supplier of both edible oil and renewableindustrial feedstock. The oil yield from wild groves is presently less than0.5 tlhaly. However. through breeding and selection. the oil yield of com-mercial plantations could reach as much as 8 tlhaly. New planting materialsalso have the capability of better oil yields with high iodine value (IV).slow height increment, and larger kernels. The oil also contains consider-able amounts of carotenoids (500-700 ppm), vitamin E (600-1000 ppm).and sterols (250--620 ppm). The oil yield of another oil palm species. E.alei/era. is approximately 0.5 tlhaly with high contents of carcrencids(700--1500 ppm). vitamin E (700--1500 ppm). and sterols (3500-4000ppm).lbe above traits could be improved through breeding and biotech-nology. Biotechnological efforts at the Palm Oil Institute of Malaysia aredirected toward the production of oil with high IV and high mO[\O\lnsatu-rated fatly acids for edible purposes and industrial uses. Isolation and ma-nipulation of the genes involved in the biosynthesis of Iauy acids are themain focus. lbe aim is to increase the efficiency of conversion of palmitate(CI6:0) to oleate (CI8:1)' Levels of palmilate and oleate are controlled bythe enzymes acyl-acyl carrier protein (ACP) rhioesterase and ~-keto acylACP synthase II. The chain termination reactions of C16:0 and CIS: I areindependent. thus paving the way for the possibility of reducing palmitatelevels by switching off the palmitoyl ACP tntcesterese gene.JAOCS 74.1451-1455 (1997).

KiMtiC!l or Transesterilication or Soybean Oil. H. Noureddini· and D.Zhu, Department of Chemical Engineering. University of Nebraska. Lin-coln. Nebraska 68588-0126.

Transesterificati<:m of soybean oil with methanol was investigated.Three stepwise and reversible reeeueos are believed to occur. The effect ofvariations in mixing intensity (Reynolds number = 3.100 to 12.400) andtemperature (30 to 70°C) on the rate of reaction were studied while themolar ratio of alcohol to triglycercl (6:1) and the concentration of catalyst(0.20 wt% based on soybean oil) wen: held constant. The variations in mix-

INFORM. Vol. 8. no. 12 (December 1997)

ing intensity appear to effect the reletion parallel to the variations in teen-perature. A reaction mechanism consisting of an initial mass transfer-con-trolled region followed by a kinetically controlled region is proposed. Theexperimental data for the lauer region appear to be a good fit into a second-Ofder kinetic mechanism. The reaction rate constants and the activation en-ergies were determined for all the forward and reverse reactions.JAOCS 74.1457-1463 (1997).

PurincalJon of 'l"Linolenle Add from BoI'1lif' Oil by a Two-Step Enzy-malic Method, Yuji Shimadaa,-, Akio Sugiharaa, MasahiroSlIibahirakib• Hiroyuki Fujitac• Hirorumi Nakand'. Tosllihiro Nagactl.Tadarnua Teraib, and Yoshio Tominagaa, aOsaka Municipal TechnicalRese:arell Institute. Osaka 536 Japan. bOsaka Institute of Technology.Department of Applied Chemistry. Osaka 535. Japan. and l"fbe NipponSynthetic Chemical Indusuy Co .. Led.. Osaka 567, Japan..

'l"Linoienic acid (GLA) was purified from borage oil by a two-step en-zymatic method. TIle first step involved hydrolysis of borage oil (GLAcontent, 22.2 WI%) with lipase:. Ps<!udomatull 5p. enzyme (LIPOSAM). Amixture of 31 borage oil, 21 watcr. and SOOO units (U) LlPOSAM was in-cubated at 350C with stirring at:KlO rpm. The reaction was 91.5':1>com-plete after 24 h. TIle resulting free fally acids (FFA) were extracted fromthe reaction mixture with a-hexane (GLA content. 22.5 wt':l>; recovery ofGl.A. 92.7%). The second step involved selective esterifICation of borage-FFA with lauryl alcohol by using Rhiwpus d<!/<!mDr lipase. A mixture con-taining 4 g bonge-FFA/lluryl alcohol (I :2. mol/mol). I g water. and !OOOU lipase was incubated It JOOc for 20 h with stirring at 500 rpm. Underthese: conditions. 74.4% ofborage·FFA was esterified. and the GLA con-tent in the FFA fraction was enriched from 22.5 10 70.2 wt'I> with a recov-ery of75.1 % of the initial content. To further elevate the GLA content. un-csterified fauy acids WCTC extracted, and esterified again in the same man-ner. 8y this repeated esterification. GLA was purified 1093.7 WI% with arecovery of 67.5% of its initial content.JAOCS 74,1465-1470 (1997).

Synthesis or (Z)-J-Huen·J·yl Butyrate In Hexane and Solvent-FreeMedium Using Mucor mltht; lind Candida antarctica Lipases. S.Bourg-Garros. N. Razafindramboa. and A.A. Pavia". Laboratoire d' Agro-biologic et Chirnle du v~g~tal, Facult~ des Sciences. 84000 Avignon.France.

(Z)-3-He~en-l-yl butyrate is an important flavor and fragrance com-pound as it represents the model of II natural herbaceous (green) note. Twoimmobilized llpases from Mucor michel (Lipozym 1M) and from Crmdicroamarctlca (Novoeym 435) were investigated for their use in the synthesisof (Z)-3-hexen-l-yl butyrate by direct esterification in e-hexane. To deter-mine optimal conditions for esterification. we examined the following pa-rameters: temperature. amount of lipase. ectdratccnct ratio. and absence ofsolvent. In a-hexane, bloconverston yields reached 95 (after 4 h) and 92%(after 6 h) for. respectively, Upozym 1M [17 (wlw reactants)] andNovczym 435 [2% (wlw reactants»). In the absence of solvent. at 6Ooe,Novozym 435-catalyz.ed esterification afforded the title compound in 80%yield. Up to 250 g (in hexane) and 160 g (without solvent) of ester wereeasily prepared. in a single operation. at a laboratory scale. in few hours.using 2% (wlw reactants) lipase,JAOCS 74. 1471-1475 (1997).

Synthesis or Coooa Butter Equi\'llient by Lipase-Catalyzed Inleresler-ificatJon In Supen:ritical Carbon Dioxide. Kuan-Ju Liu'l. Hong-MingChengi'. Rey-Cbang Changc:'. and Jei-Fu Sha.,..a,-. °lnstitute of BOlIIny.Academia Sinica. Nankang. Taipei. Taiwan 11529. bDepanment ofMarine Food Science. National Taiwan Ocean Uni~ersil)'. Keclung. Tai-wan 20224. and COcpanmcnt of See-Peed Technology. China Junior Col-lege of Marine Technology. Taipei. Taiwan III.

With supcrcritical carbon dioxide as I reaction medium. the synthesc:sof cocoa butter equivalent by interesterification with various lipases WCTCmvesdgared. Tbe study ihowcd that among \ho§e five lipases tested. lipase:IM-20 from Mucor mithei was the most effective and specific in synthe-sizing this cocoa butter equivalent product by imeresterificatioe. 1bc yieldsof cocoa butter equivalent are affected by pressure. substrate oil composi-tion. solubilily and co-solvent. The best reaction conditions ..were: reactionpmisurc at 1500 psi. uiglyceride with high content of pop (P. palmitate;0, oleate) and POQ, reaction medium with 5.0% water. and reaction rem-perature at 5O"C. 1bc major component of cocoa butter. !'OS (S. stCartlte).can be increased by 6.0% by adding I small amount of carbon dioxide. 1bcyield and melting point of the purified cocoa huller equivalent are 53.0'l0and 34.3OC. respectively.JAOCS 74. 14TI-1482 (1997).

Lipase-Catalyzed Alcoholy~is wilh Supercrili(lll Carbon DioxideE:llrlction I: rnnueece or Flow Rate. Helga Gurmleugsdottir'" and8jOrn Slvlk, Department of Food Technology. Lund Uni\·ersity. 22100Lund. Sweden.

A combined process of lipase: (E.C. 3.1.1.3) catalysis and extraction ofproduct with supercrirical carbon dioxide was studied. 'The effect of diffcr-em flow rates of the extraction fluid on the selective removal of the ethylesters (EE) synthesized in a lipase-catalyzed alcoholysis of cod liver oilwith ethanol was investigated. The faster the flow rate, the faster the ex-traction rate and the higher the recovery of EE. For example. after a 270-min extraction. the total recovery of EE was 1520 mg fOf a flow rare of 0.3liter carbon dioxide at atmospheric pressure and room temperature/min(NUmin) as compared to 250 mg when 0.Ql5 NUmin was used. The con-centration of EE in the carbon dioxide was found to decrease with increas-ing flow rate. which indicates that the rate of diffusion of EE limits theirextraction at fast flow rates. A high flow rate was found \0 result in a moreselective extraction of EE. i.e., less amounts of other lipid components pre-sent in the reaction mixture were coexuecred with the EE. Further, by in-creasing the flow rare, the equilibrium of the reaction was shifted slightlytoward ester synthesis. An increase in the flow rate from 0.015 to 0.075NUmin resulted in an approximately 10% increase in total conversion(from 73 to 82%), whereas only a negligible increase was obtained whenthe flow rate was increased further 10 O.15 NUmin.JAOCS 74, 1483-1490 (1997).

Lipase-Catalyzed AlcoholysIs with SopercritJcal Carbon DioxideExtraction 2: Phase Behavior. Helga GunnlaugsdottirD.·. Anders A.Karlssonb• and 8jOrn SivikCl• <Depanmem of Fond Technology. LundUniversity. 221 00 Lund. Sweden, and hchemical Ecology and Ecotcxr-cology, Department of Ecology. Lund University. 223 62 Lund. Sweden,

The phase behavior of systems containing ethanol plus lipid sampleswith differentllpld compositions plus carbon dioxide was studied visuallyat 4O"C and pressures ofO.OJ. 9. 15. and 23 MPa by means ofa high-pres·sure sapphire cell, The systems studied represent the main componentspresent in a lipase-catalyzed alcoholysis reaction of cod liver oil in super-critical carbon dioxide. Two phases. I vapor and a liquid phase:. were ce-served in all systems studied at supcn:ritical conditions.JAOCS 74, 1491-1494 (1997).

INFORM. VOl. 8, no. 12 (December 1997)

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1298

ABSTRACTS FROM AOCS JOURNALS

The Errect or Dietary DocO:!lahuaenoic Acid on Platelet Function,Platelet Flilly Add Composition, and Blood Coagulation in Human.s.OJ. Nelsona, P.S. Schmidra··, G.L. Bartolinia, 0.5. KelleyD. and DavidKyleb, swesem Human Nutrition Research Center, ARS. USDA. SanFrancisco, California 94129. and br.1artek Corporntion. Columbia, Macy-land 2104j.

TIle effect of dietary docosahexaenoic acid (DHA) in thc: absence ofefcosapemaenctc acid (EPA) has been stlldied infrequently in humansunder controlled conditions. This 120-d siudy followed hcallhy. adul!male volunteers who lived in the metabolic research unit (MRU) of theWestern Human Nutrition Research Center for the entire study. TIle basal(low·DHA) diet consisted of natural foods (30 en'l> fat. 15 en'l> protein.and SS en" carbohydrate). containing <SO mgld of DHA. and mel therecommended daily intake for all essential nutrients. The high·DHA (in-tervention) diet was similar except that 6 gld of DHA in the fonn of atriglyceride cootaining 4()'l, DHA replaced an equal amount of saffloweroil in the tHlsal diet. The subjects (ages 20 to 39) were ....ithin -10 to+2<I'k of idc:al body weight. nonsmoking. and DOl allowed alcohol in the:MRU. Their exercise level was constant. and their body weights weremaintained within 2'>' of entry level. They were initially fed the tow-DHA diet for 30 d. On day 31. six subjects (intervemion. group A) wereplaced on the high·DHA diet; the other foor subjects (controls, group B)remained on the 10w·DHA diet. Platelet aggregation in platelet-richplasma was determined using ADP. collagen. and arachidonic acid. Nostatistical differences could be detected between the amount of agonistrequired to produce 50% aggregation of platelet-rich plasma before andafter the subjects consumed the high·DHA diet. The prothrombin time.activated partial thromboplastin time, and the antithrornbin-Hl levels inthe subjects were determined. and. again. there were no statistically sig·nificant differences in these three parameters when their values werecompared before and after the subjects consumed the hlgh-Dl+A diet. Inaddition. the ill vivo bleeding times did not show any significant differ-ence before and after the subjects consumed the high-Dl+A diet (9.4:t3.1 min before and 8.0:t 3.4 min after). Platelets from the volunteers ex-hibited more than a threefold increase in their DHA content from 1.54 :t0.16 to ~.48:t 1.21 (wt%) during the DHA feeding period. The EPA con-tent of the subjects' platelets increased from 0.34 :t 0.12 to 2.67 :t 0.91(wt%) during the high-DHA diet despite the absence of EPA in the sub-jects' diets. The results from this study on blood clotting parameters andin vitro platelet aggregation suggest that adding 6 g/d of dietary DHA for90 d to a typical Western diet containing less tban 50 mgfd of DHA pro-duces no observable physiological changes in blood coagulation. plateletfunction. or thrombotic tendencies in healthy, adult males.Upids 31. 1129-1136 (1991).

The Err«t or l)ietary DocosabuHnolc Acid on Plasma Lipoproteinsand TIssue Fatty Acid Composition In Humans. GJ. Nelsotf',·. P.e.Schmidr", G.L. Banolinio. D.S. Kelley<!, and David Kyleb. QWestemHuman Nutrition Rcsc:art:h Center. ARS. USDA. San Francisco, Calif or-nia 94129. and ~artck Corporation, Columbia, Maryland 21()4S.

Nonnal, bealthy male vclumeers (II = 6) wen: fed diets [high ecccse-hcxac:noic acid·DHA] cootaining 6 gfd ofDHA for 90 d. The stabilization(low·DHA) diet contained less than 50 mg/d of DHA. A control group(n .. 4) remained on the low·DHA diet for the duration of the study (120d). Blood samples were dra ....n on study days 30 (end of the stabilizationperiod). 75 (midpoint of the intervention period), and 120 (end of the in-tervention period). Adipose tissue (An samples wen: taken on days 30 and120. The plasma cholesterol (C), low density lipoprotein (LDL)·C andapolipoproteins (apo) [AI. B. and lipoprotein (a)] were um:hanged afler 90d, but the triglycerides (TAG) wen: reduced from a mean value of 76.67:t24.32 to 63.83 :t 16.99 mg/dL (n = 6. P < 0.007 using a pain:d r-test) andthe high density lipoprotein (HDL}C incn:ascd from 34.83:t 4.38 mg/dLto 37.83:t 3.32 mgfdL (n = 6, P < om 7 using a paired I·test). The controlgroup showed no significant reduction in plasma TAG levels. Apo-E, how·

ever. shewed a marked increase in the volunteers' plasma after 90 d on thehigh·DHA diet. from 7.06:t 4.47 mg/dL on study day 30 10 12.01 :t 4.96mg/dL on study day 120 (P < 0.002 using a paired r-tesr). The control sub-jeeu showed no significant change in the apo-E in their plasma (8.46:t2.90 on day 30 VI. 8.59:t 2.97 on day 120). The weight percentage ofplasma DHA rose from 1.83:t 0.22 to 8.12 :t 0.76 after 90 d on the high-DHA diet. Although these volunteers were eating a diet free of eicosepen-taenolc acid (EPA). plasma EPA levels rose from 0.38:t 0.05 to 3.39:t 0.52(wt%) after consuming the high·DHA diet. The fatty acid composition ofplasma lipid Iractions-c-cbelesterol esters. TAG. and phospholipid-showed marked similarity in the enrichment of DHA. about 10%, after thesubjects consumed the high·DHA diet. The DHA content of these plasmalipid fractions varied from less than 1% (TAG) to 3.5% (phospholipids) atbaseline. study day 30. EPA also increased in all plasma lipid fractionsafter the subjects coosumed the high·DHA diel. There .....ere no changes inthe plasma DHA or EPA levels in the control group. Consumption of DHAalso caused an increase in AT levels of DHA. from 0.10:t 0.02 to 0.31 :t0.07 (WI"') (" • 6, P < 0.001 using a paired "test), but the amount of EPAin their AT did not change. Thus. dietary DHA ....illiower plasma TAGwitnout EPA, and DHA is retrcconverted 10 EPA in significant amounts.Dietary DHA Ippean 10 enhance apo-E synthesis in the liver. Itlppearsthat DHA can be. safe and perilaps beneficial supplc:mcnllO human diets.Upids 31, 1137-\146 (1997).

Panenalie 811e Salt·Dependent Lipase Activity in Serum or Nor-moIlpidemk Patients. Nathalie Caillol., Eric Pasqualini, Eric Mas, Annevsteue, Allin Verine, and Dominique Lombardo". INSERM U·260,13385 Manc:iI1e ~ 5. France.

Bile saJt-dependent lipase (BSDL, E.C. 3.\.\.-) is a digestive enzymesecreted by the pancreatic acinar cell. Once in the duodenum, the enzyme,upon activation by primary bile salts. hydrolyzes dietary lipid es~rs suchas cholesteryl esters and Hpld-soluble vitamin esters. This enzyme is par-tially transferred from the duodenum or pancreas 10 the circulation whereit has been postulated to exert a systemic action on atheroma-generatingoxidized·low density lipoprotein (LDL). In the pres-ent study, sera from40 healthy normolipidemic volunteers were used to investigate the possiblelinkage between circulating BSDL. lipids. and lipoproteins. We showed,firstly, Ihut pancreatic-like BSDL activity can be detected in these serums.Secondly, BSDL activity increased significantly with the level of LDL-cholesterol and was also positively linked to tbe serum concentration ofApo B 100 and Apo A·1. Thirdly. we also established that BSDL was esse-ciated with LDL. in pan by a specific interaction with Apo B 100, while nointeraction was found with Apo A·1. No linkage with other recorded pa-rameters (triglycerides, phospholipids, and high density lipoprotein-cholesterol) was detected. Because an increase in LDL-cholesterol repre-scots an important risk factor for atheroma. the concomitant increase inBSDL, which can metabolize atherogenic LDL, suggests for the first timethnt this circulming enzyme may exert a positive effect against atheroscle-rosis.Upids 32,1147-1153 (1997).

Epicoproslanol Found In Adipocere rrom Five Human Autopsies.Junko Adachill, •. Yasuhiro uenos, Atsukc Miwall. Migi ....a Asanoll,Akiyoshi Nishimurab, and Yoshitsugu Tatsunoll. lIDc:parunent of LegalMedicine, Kobe University School of Medicine. Kobe 650, Japan. andbShiga University of Medical Science, Dhtsu 520-21. Japan.

Adipocere formation is wellll:nown as a la~r postmortem change. Wecollected adipocere from five male victims which had been submergedunder the sea Of f~sh water fOf I moo 10 4 yr. Fresh subcutaneous fat of.male victim who died from a cerebral contusion was used as the control.The samples wen: homogenized, and the lipids were extracted with chlo-roform and methanol follo .....ed by injection into a gas chromatograph anda gas chromatograph-mass spectrometer. We detected hydroxy fany acids(lO-hydroxyoctadecanoic acid and IO-hydroxyhexadecaooic acid) as wellas 100ketooctadecanoic acid in adipocere. but not in the control. In addi-tion. we found fOf the first time a cholesterol-related peak with a molecularion of 388 in adipocere and identified it as epicoprostanol. suggesting notonly oxidation but also reduction had occurred during the formation ofadipocere. In addition. we showed the nme-cccrse of epicoprostanol accu-mulation. The relationship between the time of adipocere formation andthe chllnlCterislic lipid composition is discussed.Upids 32. 115}-1160 (1997).

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1299

Preventlen or Ischemia-Induced Cardiac Sudden Death by n-3Polyunsaturated Fally Acids In Dogs. George E. Billmanu, Jing X.Kangt'. and Alexander Leati'··, uDepanmenl of Physiology, The OhioState University, Columbus. Ohio 45210. and bocpanmenlS of Medicine.MassachuscIIs Gcna1.1 Hospital. BosIOl'l. Massachuscns 02114 and Har-vard Medical School, Boston, Massachusetts 02115.

The objective of this s!lId)' was to obtain funclional infonnation as-sociated with the prevention by n·) poJyunsawllIled fany acids (PUFA)of ischemia-induced Iatal cardiac ventricular arrhythmias in the intact.conscious. exercising dog. Thirteen dog! susceptible 10 ischemia-in-duced ventricular fibrillation were prepared surgically by ligation oftheir anterior descending lefl coronary artery and placement of an in-flatable cuff around their left circumflex artery. After 4 wk of recovery.exercise-plus-ischemia tests were performed without Ind then with In

intravenous infusion of an emulsion of free n-3 PUFA just prior to oc-cluding the left circumflex artery while the animals were ronning on Itreadmill. One week liter the exercise-plus-ischemia test WIS repeatedbut with a control infusion replacing the emulsion of n-3 PUFA. 1lIc: in-fusion of ttle free n-3 PUFA in quantities of 1.0 to 10 g prevented ven-tricular fibrillation in 10 of the 13 dogs tested (P < OJl(5). apparentlywithout estcrification of the PUFA into membrane phospholipids. Theantiarrhythmic effect of ttle n-3 PUFA was associated with slowing ofthe hean rite, shortening of the QT-interval (electrical action potentialdUrition). reduction of left ventricular 'Y5tolic pressure. and prolonga-tion of ttle electnx:ardiographic atrial-ventricular conduction time (P-Rinterval). These effccts are comparlble with those we have reported instudies with cultured neonatal ral cardiac myocytes.Upids 32,1161-1168 (1997).

Biliary Excrftlon of DoUchols and ~-Huosamlnldase-Errect ofEtha~ and Glucagon, Kari Humaloja, Mikko Salaspuro·, and Risto P.Roine, Research Unit of Alcohol Diseases. Helsinki Uni\'ersity CentralHospital, Helsinki, Finland.

Aleohol has been reported 10 increase the urinary excretion ofdolichols, and urinary dolichols are suggested 10 be deri\'ed from the lyse-somes of the renal eeus. In the present study we examined the effects ofa1eohol and glucagon on the biliary excretion of dolichols in rats. Chronicethanol treatment decreased both biliary dolichol and ~-hexosaminidaseexcretion. 1lIc: absolute amount of dolichol excreted into the bile correlatedhighly significantly with the absolute amount of biliary ~llCJiosaminidase.Our results indicate that biliary dolichols are-at least in part-<leri\'edfrom hepatic lysosome!. Decreased biliary dolichol output during chronicalcohol administration suggests that urinary and biliary dolichol excretionsare regulated independently of each other.Up/tis 32. 1169-1172 (1997).

Effect of Curcumtn and Capsaicin on Arachidonic Acid Metabolismand Lysosomal Enzyme secreuee by Rat Peritoneal Macrophages.Bina Joe and B.R. Lokesh·, Department of Biochemistry and Nutrition.Central Food Technological Resean:h Institute. Mysore-570 Oil. India.

The inflammatory mediators secreted by macroph.age$ play an impor-tant role in autclmmcne diseases. Spice components. such as curcumlnfrom turmeric and capsaicin from red pepper, are shown to exhibit antiin-flammatory properties. The influence of these spice components on arachi-donic acid metabolism and secretion of lysosomal enzymes bymacrophages was in\·estigated. Rat peritoneal macrophages preirw::ubatedwith 10 ~ curcumin or capsaicin for I h inhibited the incorporation ofarachidonic acid into membrane lipids by 82 and 76~: prostaglandin qby 45 and 48~; leukOlriene B4 by 61 and 46~. and leukouienc: C4 by 34and 48~, respectively, but did not affect the release of arachidonic acidfrom macropbges stimulated by phorbol myristate acetate. However. thesecretion of 6-keto PG FIQ. was enhanced by 40 and 29% frommacrophlge5 prelncubated with 10]lM cun;:umin or capsaicin. respec-ti,·ely. as compared to those produced by control cells. Curcumin and cap-picin also inhibited the secretion of collagenase. elastase. andhyaluronidase to the maximum extent of 57, 61, 66~, and 46. 69. 67~, re-specti\'ely. These results demonstrated !hat curcumin and capsaicin cancontrol the: release of innammatory mediators such as ekosanoids and hy-drolytic enzymes secreted by macrophages and thereby may exhibit anti-inflammatory propertie$.Upitis 32, 1173-1180 (1997).

Generation and Remodeling or Highly Polyunsaturatoo MolecularSpecies or Rat Hepatocyte Phospholipids. Patricia C. Schmid, IlonaSpimro va, and Harald H.O. Schmid·, 1lIc: Hormc:llnstitute. Uni"cnity ofMinnesota. Austin. Minnesota.

Freshly isolated rat hcpatocytes were incubated foc 20 min with [U-14C)glycerol in the presence or absence of unlabeled linoleic (18:2n-6).I1*:hidonic (20:4n-6). or docosaheXlICnoic (22:6n-3) acid, added as albu·min complex in l~ ethanol. Most of the radiOlCli\'ity (-95~) recoveredin hepatocyte lipids was present in phosphatidylclloline (PC), phos-phatidylethanolamine (PE). and triacylglycerol (TAG). The presence ofexogenous fatty acids resulted in 0) highet- incorporation of [U-I4cJglyc-ercl. (ii) higher percentage of label in TAG. and (iii) enhanced formationof PC and FE molecullr species bearing the exogenous fatty acid at boththe Sri-I and srI-2 positions of glycerol. In each case. these molecularspecies contained 60 to 7~ of the label in that lipid class. Further incuba-tion of the cells for 40 and 80 min in the absence of labelfd substrate andexogenous fally acids resulted in a redistribulion of label among PC andPE molecular species due to deacylatioo-reacylation at the sn-I positionof glycerol.Upitis 12.1181-1187 (1997).

Anal)'SIs of the Sftd Oil of Htisltritlsifvanii (Olacaceae}-A RlchSoun:e ora No~el CI8 Am,'1mic Fatty Acid. Volker Spittef'1-·, WernerTombergb, Rudolf Hartmannb. and ReiDC1'AkhholzC. QFacuity of Phar-macy, Federal University of Rio Grande do Sui (UFRGS), 90.6\0.000Porto AlegrelRS. Brazil. bJ>bysioIOSisch-Chemiscnes lnstinn. Uni\'cnititBonn, 53115 Bonn, Germany, and CNO\'Irtis Pharma AG. Research. COleTechnology ArealAnalytics, CH-4002 Basel. Switzerland.

Besides some usull fallY acids (FA). two conjugated ene-yneacetylenlc FA [lrons-IO-heptadecen-8-ynoic acid (pyrolic acid) (7.4~).Ind Irorls-11-octadecen-9-ynoic acid (ximenynic acid) (3.5~»), a novelene-yne-ene acetylenic FA [ds- 7. Irons-II-octadecadiene-9-ynoic acid(beistcric acid) (22.6'1>)), and 9,IQ-epoxysteark acid (0.6'1» could be iden-tified in the seed oil of Heijluia $iI"anii (Olacaceae). Two further conju-gated ecetylenic FA [9,II-octadecadiynoic acid (0.1 '1» and 13-octadecene-9,1 l-diynoic acid (0.4'1>)] .....ere identified tentatively by their mass spectra.Tbe FA mixture has been analyzed by gas chromatographylmass spectrom-etry (GClMS) of their methyl ester and 4.4-dime!hyloxazoline derivatives.The structure of belsterio acid was elucidated after isolation ";0 preparati\'esilver ion thin-layer chromatography and by various spectroscopic methods[ultraviolet: infl'lU't'd; I H. 13C nuclear magnetic resonance (NMR): I H_I Hand I H_13C correlation spectroscopy). To determine the position of theconjugated ene-yee-eee system. the NMR spectra were also measured afteraddition of the lanthanide shift reagent Resclve-Al EuFODni. Further-more. the triyglyceride mixture was analyzed by high-temperature GC andhigh-temperature GC coupled with negative chemical ionization MS. Aglass capillary column coated with a metboxy-rermineted 500%-diphenyl-5O%-dimethylpolysiloune was used for the separation of the triacylglyc-erol (TAG) species. No evidence of decomposition oflbe TAG species con-taining conjugated ene-yne-ene FA was observed. Twenty-six species ofthe:separated TAG were identified by means ofthe:ir abundant quasi molec-ular ion [M - H)-and thc:ir corresponding carboxylate anions (RCOOr ofthe: fatty acids, respecuvely. 1lIc: major molecular species of the: TAG werefound to be 16:0118: 1/18: I, 16:0118:1/18:3 (heisteric acid), 17:2 Ipyrulicacid)llg:1I18:1. 18:1118:1118:3 (heisteric acid). The TAG containingacetylenic FA !JIoIo,'ed an unexpected increase of the: retention time in com-parison 10 the TAG containing usual FA. thus making the prediction of theelution order of lipid $IJIIples contlining acetylenic FA difficultUp/tis 32. 1189-1200 (1997).

Lipid Spedf\clty and LoeatkHi of the Sterol Caniu Protein-2 FattyAdd-Binding Site: A Fluol"e5«l\C't DIsplacement and Energy Trans-fu Study. Andrey Frolovt'. Kimberly Miller'. Jeffrey T. Billheimerb.Tae-Hyeon CtKf'. and Friedheim Schroeder"-·, QDepamnem of Physiolo-gy and Pharmacology. Texas A.t.M University. 1VMC. College Station,TexIS 77843-4466. Ind bCardiovlscular Department. DuPont MerckPharmaceutical Company Experimental Station 400-3231. Wilmington.Delaware 19898-0400.

Althoup it was recently recognized that sterol carrier protein-2 (SCP-2) interacts with fatty acids. little is known regarding the specificity ofSCP-2 for long-chain flny acids or br1Inched-chain fally-acid-likemolt'alles. Ukcwisc the location of the fany-acid binding site within SCP-

INFORM. \,bI. 8, no. 12 (December 199n

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Evaluating Add and Base Catalysts in Ihe Methylation of Milk lindRumen Fatty Acids with Special Emphasis on Conjugated Dienes andTotal trans Falty Adds. John K.G. KramerD,·, vivek Fellner', MichaelE.R. DuganO, Frank D. Sauer". Magdi M. Mossobah, and Martin P.Yuraweczb, aCenter for Food and Animal Research. Research Branch.Agriculture and Agri-Food Canada. Ouawa. Ontario, Canada, KIA OC6,and eus. FDA. Center for Food Safety and Applied Nutrition. Washing-ton. OC 20204.

Milk analysis is receiving increased enennon. Milk contains conju-gated octadecadienoic acids (] 8:2) purported 10 be anucercincgemc. lowlevels of essential fany acids. and trons fatty adds that increase when es-sential fatty acids are increased in dairy rations. Milk and rumen fatty addmethyl esters (FAME) were prepared using several acid- (He1. SF3. acetylchloride. H2S04,l or base-catalysts (NaOCH3' tetramethylguanidine. dia-zomemaee), or combinations thereof. All acid-catalyzed procedures re-sulted in decreased cis/trans (69c.lll-18:2) and increased lrons/rrans(691.111-18:2) conjugated dienes and the production of allyllc methoxy ar-tifacts. The meihoxy artifacts were identified by gas-liquid chromatogra-phy {GLC}mass speceosccpyrte base-catalyzcd procedures gave no iso-merization of conjugated dienes and no methoxy ertlfacts. but they did nottransesterify N-acyl lipids such as sphingomyelin. and NaOCH3 did notmethylate free fatty acids. In addition. reaction with tetramethylguanidineccextrected material with hexane that interfered with the determination ofthe short-cham FAME by GLe. Acid-catalyzed methylation resulted in theloss of about 12% total conjugated dienes. 42% recovery of the 69c.lll·18:2 isomer. a fourfold increase in 691.111-18:2. and the formation ofmethcxy artifacts, compared with the base-catalyzed reactions. Total milkFAME showed significant infrared (IR) absorption due to conjugated di-enes at 985 and 948 cm-I. The IR determination of total Irons content ofmilk FAME was not fully satisfactory because the 966 cm-[ trans bandoverlapped with the conjugated diene bands. IR accuracy was limited bythe fact that the absorptivity of methyl elaidate. used as calibration stan-dard. was different from those of the other minor lraltS fally adds (e.g .• di-enes) found in milk. In addition. acid-catalyzed reactions produced inter-fering material that absorbed extensively in the Irons IR region. No singlemethod or combination of methods could adequately prepare FAME fromall lipid classes in milk or rumen lipids. and not affect the conjugated di-enes. The best compromise for milk fatty acids was obtained withNaOCH3 followed by HCI or BF3. or diazomethane followed byNaOCH3. being aware that sphingomyelins are ignored. For rumen sam-ples. the best method was diazomethane followed by NaOCH3'Lipids 32. 1219--1228 (1997).

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ABSTRACTS FROM AOCS JOURNALS

2 is unresolved. A fluorescent cis-parinaric acid displacement assay wasused to show that SCP-2 optimally interacted with 14-22 carbon chain li-pidic molecules: polyunsaturated fatty acids> monounsaturated. saturated> branched-chain isoprenoids> brunched-chain phytol-derived fatty acids.In contrast, the other major fatty-acid binding protein in liver. fatty-acidbinding protein (L-FABP). displayed a much narrower carbon chain pref-erence in general: polyunsaturated fatty acids> branched-Chain phytol-de-rived fatty acids> 14- and 16-carbon saturated > branched-chain iso-prenoids. However. both SCP-2 and L-FABP displayed a very similar un-saturated fatty-acid specjflciry profile. The presence and location of tbeSCP-2lipid binding site were investigated by fluorescence energy transfer.The distance between the SCP-2 Trp50 and bound cis-parinaric acid wasdetermined to be 40 A. Thus. the SCP-2 fatty-acid binding site appeared tobe located on the opposite side of the SCP-2 Trp50. These findings notonly contribute to our understanding of the SCP-2 ligand binding site butalso provide evidence suggesting a potential role for SCP-2 and/or L-FABPin metsbofistn of branched -chain fauy acids and isoprenoids.Upids 32. 1201-1209 (1997).

Elution Faetera of Synthetic Oxotrlacylglycerols as an Aid in Identin_cation of Peroxidi:ted Natural Trtecylglycercls by Reverse-PhaseHigh-Performance Liquid Chromatography with Eleclrospray MassSpectrometry. Olli SjOvsllo•b. Amis Kultsistl·-. Lajns Maraia. and JohnJ. Myher". aBanting and Best Department of Medical Research. Universi-ty of Toronto. Toronto. Canada M5G 1L6. and bDcpanment of Biochem-istry and Food Chemistry. University of Turku, F1N-20014 Turku. Fin-tand.

Selected elution factors were determined for model oxoutscytgtycerotsas an aid in identification of the percxidation products of natural triacyl-glycerols by reverse-phase high-performance liquid chromatography(HPLC) with electrcspray mass spectrometry (LClESfMS). For this pur-pose synthetic triacylglycercls of known structure were convened to hy-droperoxides. hydroxides. epoxides. and core aldehydes and their dinitro-phenylhydrazones by published procedures. The oxorriacylgtycercls wereresolved by normal-phase thin-layer chromatography and reverse-phaseHPLC. and the identities of the oxotriacylglycercls confirmed byLClESfMS. Elution factors of oxotriacylglycerols were determined in re-lation to a homologous series of saturated rriacylglycerols. ranging from24 to 54 acyl carbons. and analyzed by reverse-phase HPLC, using a gra-dient of 20-80% isopropanol in methanol as eluting solvent and an evapo-rative light-scattering detector. It was shown that the elution times variedwith the nature of the functional group and its regiclocauon in the macyj-glycerol molecule. A total of 31 incremental elution factors were calcu-lated from chromatography of 33 oxygenated and oonoxygenated macyl-glycerol species. ranging in carbon number from 36 to 54 and in double-bond number from 0 to 6.Upids 32. 1211-1218 (1997).

Effects of High Pressure and Temperature on Micelle Formatinn orSodium Deoxycholate and Sodium nocecytsutrate. Michiko Kate,Shogo Ozawa. and Rikimaru Hayashi", Laboratory of BiomacromolecularChemistry, Graduate School of AgriCUlture, Kyoto University. Kyoto 606-01. Japan.Lipids 32. 1229-1230.

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INFORM, Vol. 8, no. 12 (December 1997)