214

BIOTECHNOLOGY

Steroid transformations by microbial systemsA great deal of work has been car-

fiiied QUIon production of fatswith many yeasts and molds. At pres-ent, however, industrial applicationshave been limited. Nonetheless. sincethe worldwide demand for oils isincreasing faster than production fromconventional plant sources. the dis-'tinct possibility exists thai more defi-nite commercialization of microbialsystems will be undertaken.

Yeasts and molds may also proveto be useful commercial sources forsterols. While the conditions for opti-mal production have 10 be established,there have been considerable develop-ments on microbial transformations ofindividual steroid subsrrares into use-ful steroid drugs and hormones ortheir useful intermediates. A variety ofsteroids are widely used as antiinffam-matory, diuretic. anabolic, contracep-tive. anti androgenic. prosgesrauonaland anticancer agents as well as inother applications.

The importance of microbial trans-formations was realized for the firsttime in 1952 when Murray and Peter-son of Upjohn Company patented theprocess of I l u-hydrexylation of pro-gesterone by a Rhizopus species. Thechemical methods for preparation ofprogesterone from widely availableplant steroids, e.g. diosgenin (from theMexican yam) and stigmasterol (fromsoybeans), were well standardized. Inaddition. conversion of progesteroneto corticosteroids requires introduc-tion of a specific oxygen functionwhich can be achieved by a combina-tion of chemical methods only withgreat difficulty and at a high cost. Assuch the single-step high-yieldingmicrobial process immediately wasconsidered for industrial exploitation.Since then microbial reactions for thetransformation of steroids have prolif-erated and specific microbial transfor-mation steps have been incorporatedinto numerous paruat syntheses ofnew steroids for evaluation as drugsand hormones. The synthetic protocolof a steroid drug or hormone, whichgenerally comprises chemicals as wellas microbial steps. is so designed thatthe microbial transformation is con-

"Processes for the commercial conversion ofsterols to 17-ketosteroids by mutants ofMycobacteria and Corynbacteria have beendeveloped and are in practical use."

typical microorganisms that are indus-trially used for I tu-hydroxylauon areRhizopus nigricans and Aspergillusniger whereas strains of Cunning-hamella blakesleena and Curvularialunata are employed for II~-hydroxy-tenon. e.g. in the transformation ofSubstance S to hydrocortisone. 16a-Hydroxylation. which has becomeimportant for production of the steroidhormone analogue triamcinolone. isbest achieved commercially by Strep-tomyces roseocbromogrnes. Scrutinyof the structural features of the widelyused corticosteroids reveals that all ofthem contain the .11-double bond. Thisdehydrogenation step is best achievedby microbial biotechnology and thefrequently used microorganism forthis purpose is Arthrobaaer simplex.

Although the efficient removal ofthe side chain of cholesterol by achemical method was reported in 1975.a more promising approach employingmicroorganisms has been applied tovarious sterols. Processes for the com-mercial conversion of sterols to 17-

kerosterotds by mutants of Mycobacte-ria and Corynbacteria have beendeveloped and are in practical use. The17-ketOSleroids viz androst-4-ene-3.l7-dione (androst-enedione) or andros-ta-I.4-diene-3.I7-dione are usefulproducts for chemical modification tofinished products or intermediates.Moreover, chemical conversion ofthese l7-ketosteroids to pregnane ana-logues by reconstruction of the preg-nane side chain has replaced partiallythe use of diosgenin and solasodine asbase materials. A partial side-chaindegradation of sterols to 2O-carboxylicacid structures also has been developedand is in commercial use. Selectivecleavage of the acetyl side chain ofpregnanes by microorganisms to the C-19 steroids also is important commer-cially as 16-dehydropregnenolone

The general uses of microbial sys-tems in biotechnology are well rec-ognized but specific applications inthe oils and fats industry have beenlimited. AI the request of INFORMAssociate EditorJ.B.M. Rattray. S.B.Mahato of the Indian tnstitute ofChemicaf Biology. 4, Raja S.C. Mul-lick Road, Jadavpur, Calcutta 700032, India, has prepared the follow-ing article to illustrate the great ver-satility of microbial systems in theproduction of commercially \'aluablesteroids for the pharmaceuticalindustry.

ducted at the terminal or near terminalstep. According to a present estimate,the production of steroid drugs andhormones to the value of $400 millioninvolves biotechnological procedures.

Biotechnology applications dealingwith steroids may be classified broad-ly into two categories: (a) processingraw materials into useful intermedi-ates for general steroid production,

e.g , microbial hydrolysis of plantsteroid saponins to aglycones andmicrobial degradation of widelyoccurring sterols to useful steroidintermediates; and (b) transformationof specific steroid intermediates todesired products, e.g. microbial con-version of hydrocortisone to its .1'-dehydrogenated analogue. pred-nisolone, a highly selective antiin-Ilammatory commercial product.

Industrial applicationsThe ttc-. Ilfj- and 16a-hydroxyla-lions. 6'-tlehydrogenation and side-chain degradations are now exclusive-ly achieved in the steroid industry bymicrobial transformations. The threehydroxylations and d'-dehydrogena-tion are used in manufacture of coni-cosrerolds and their analogues. The

INFORM. \obi. 2. no. 3 (March 1991)

215

BIOTECHNOLOGY

acetate (l6-DPA) or progesterone easi-ly is derived chemically from widelyavailable steroid sapogenins or stig-rnasterol. The synthesis of AI-dehy-drcresrolactcne, a useful anticancerdrug. is best achieved by microbialdegradation of progesterone usingCylindrocorpon radicicola, Whiledegradation of the phytosterol sidechain proceeds via Il-oxidolion. thecleavage of the acetyl side chain ofprogesterone as well as the laetonationhave been shown to involve Baeyer-Villigcr-type oxidation.

Some contraceptives are producedby total chemical synthesis. However,a microbial stereospecific reduction ofthe 17-keto function is advantageouslyperformed for the introduction of thenecessary chirality. This microbialketone reduction also is applied forthe production of testosterone fromendrosr-e-ene-a.t r-dione by Saccha-romyces sp. Another microbial pro-cess which is used in the Soviet Unionand by Sobering AG is the .o.>-3j}-alco-hoi dehydrogenation.

Besides the above microbial pro-cesses which are in industrial use.other potential microbial methodshave been developed in recent yearswhich have potential for use as alter-native procedures. Microbial 90.·hydroxylation of androst-a-ene-S.t r-dione has opened up another means ofpreparation of .o.91I1J·intermediateswhich are subsequently transformedchemically to 9a.-fluorocorticoidswithout the traditional microbial II-hydroxylations. Biodegradation of thesterol fractions obtained as industrialby-products to the intermediate prod-ucts of steroid drug synthesis isimportant commercially. Mutants ofMycobacteria recently have beenshown to convert the sterol fraction ofa deodorizer condensate and the sterolfraction from tall oils from paper pulpmanufacture to 22-hydroxy-23.24-bis·norchola-t.a-dien-S-one and endrosta-I .a-dtene-S, l7-dione respectively inappreciable yields. The sterol fractionof the deodorizing condensate pro-duced from the wastes of fat industrycontained sitosterol, campesterol.brassicasterol, stigmasterol as well ascontaminated cholesterol while that ofthe tall oil was a mixture of sitosteroland campesterol.

Stigmasterol

O~

Microbialdegradation

•I ChomI<a't·...

Progesterone-» 0

Chom." . r+\steps~

o

DIosgenin

O~HOI Miaoblalt hydroxylation

11a..()H.Prognlerone

~

_ 0 Chemical

"...--o

"_.~OHdehydro.- HOgeoation--o

Chemical

"'"I Chemicalt steps

Pntc:Inlsolone

_~OHdeh~ HO ._00

""''''''-o

FIG. 1. Microorganism involvements in steroid production.

An efficient conversion of choles-terol to l7-hydroxysteroids by the useof an external proton donor has beendeveloped. The method involves incu-bation of cholesterol with a Mycobac-terium sp. in the presence of oneequivalent of hydroquinone or oneequivalent of ascorbic acid to yield thedesired product.

and intact microbial cells and immobi-lized cells. Much of the recent interestcenters on the development of pro-cesses using immobilized cells forindustrial exploitation. The industrial-ly used microbial steroid transforma-tions are based on enzymes with lowstability and as such the immobiliza-tion technique is employed for immo-bilization of intact cells. Microbialcells can be immobilized by covalentattachment to or adsorption on solidsupports. entrapment in polymericgels. or by encapsulation and cross-linking. The perfonnance of the wholecells in steroid transformation canoften be improved by using immobi-lization techniques.

Several encouraging results havebeen obtained. Thus, sequential con-version of ccrtexclone to prednisoloneby immobilized mycelia of Curvutar-ia lunata and immobilized cells ofArthrobacter simplex has been report-

Auernative processesThe commercial success of microbialsteroid rranstormeuons so far has beenachieved by nourishing vegetative cellcultures. However, developmentalstudies on several alternative process-es have been conducted for the pur-pose of improving product yield, con-trolling side reactions. simplifyingworking procedures and therebyreducing product cost. These enema-rive processes include employment ofcell-free enzymes. immobilizedenzymes. spores. immobilized spores,

INFORM, Vol. 2, no. 3 (March 1991)

216

BIOTECHNOLOGY

'There is continuing interest/or improvement inthe yields of desired metabolites in.microbialtransformation. by the use of enzyme inhibitors."

ed. Immobilized living mycelia of C.lunata having a high 11~·hydro:c.yla-tion activity were prepared by in situgermination of spores entrapped inphoro-crosstinked resin gels of a suit-able network structure. Acetone driedcells of A. simplex having an inducedsteroid 6'-dehydrogenase activity aJsowere entrapped with photo-crosslink-able resin polymers and used for 0'-dehydrogenation of hydrocortisone 10prednisolone. The production of pred-nisolone from conexotcne wasachieved convenienliy by the ccmbi-nation of sequential steps, Ill!-hydrcxylatton and subsequent .1'-dehydrogenation. Each immobilized

microbial cell system was stable andcould be used repeatedly for thesequential reactions. A new techniqueinvolving the use of an airlift loopreactor for the transformation ofsteroid by immobilized cells has beenpublished. The pcrfonnance of immo-bilized Arthrobacter simplex for the,6.1-dehydrogenation of hydrocortisonein the airlift reactor was comparable tothat in a stirred-tank reactor. A tech-nique of immobilization of cells byadhesion on glass either by coating thesupport with colloidal particles ofhydrous alumina or by pretreating thecells with aluminum has beendescribed. The use of glass slides on amodel support has shown that a sin-gle. dense and regular layer of immo-bilized cells is achieved. The quantityof immobilized cells was estimated tobe approximately 7 x 101 cells/em'.Immobilization on glass beads orglass wool packed as a bed in a col-umn also was successful. The use of amicroreactor with the immobilizedbacteria as biocatalyst demonstratedthe feasibility of repeated use of themicroorganism.

NC\4'er techniquesA novel method for improved steroid,6.'-dehydrogenation using heat-driedbacterial cells has been reponed. The

INFORM. Vol. 2, no. 3 (March 1991)

steroid .1,1-dehydrogenase was stableat high temperature and other enzymeswere inactivated. Thus .1,1-steroidswere produced by heat-driedArthrobacter simplex cells. The high-est rates of ,6.'-dehydrogenation(>97%) were observed for cells driedat 40 - 6O"C. The method of nanspo-son mutagenesis has been used for theproduction of therapeutically usefulsteroids in good yield from bile acids.The high yields are obtained by inacti-vation of steroid degrading enzymesby the introduced mutation. There iscontinuing interest for improvement inthe yields of desired metabolites inmicrobial transformation by the use of

New steroid metabolitesAlthough the introduction of newsteroids into commerce is now limit-ed, there are continuing reports onproduction of new steroids by micro-bial transformation as this techniquecan conveniently be exploited forpreparation of steroid analogueswhich are otherwise inaccessible. Anovel microbial transformation ofdehydrochlolic acid by Streptomycesgelaticus into steroid dimers contain-ing a benzene ring has been reponed.The microbial production of a novelandrostane analogue. l6a-methoxy-l7~-hydroxyandrosta-1 ,4-dien-3-oneby fermentation of 16-dehydropreg-nenolone with a strain of Arthrobactersimplex has been described. A newbile acid metabolite, 3a,15p-dihy-droxy-5~-cholanic acid was producedin good yield by transformation oflithocholic acid by the fungus Cun-ninghamella btakesteena, Hydro-phobicity measurement and choles-terol solubilization studies showedthat this new metabolite was as effec-tive as ursodeoxycholic acid in choles-terol solubilization. Two new preg-nane analogues, 14-hydroxy-5a-preg-nene-3.6,20-trione and Ila,14-dihy-droxy-4-pregnene-3,20-dione. ofpotential therapeutical interest havebeen reponed to have been producedas metabolites of progesterone by aBacillus species. Fennentation of 17-hydroxyprogesterone with the sameorganisms yielded a new pregnaneanalogue, 6 p, 17.20 u-rrthydrox-ypregn-4-en- 3-one.

The isolation on methyl 3-hydroxy-9-oxo-9, 10-seco-23,24-dinor-l, 3.5( 10)-cholatrienoate by microbialdegradation of cholesterol and sisros-terot by a mutant of Mycobacterium[ortuitum has been described. Thepreparation of the interesting metabo-lites 9-hydroxy-3-oxo 23,24-dinor-4,17(20)-choladien-22-al and 9.22-dihy-droxy-23.24-dinor-4-cholen-3-onefrom a sitosterol bioconversion alsohas been achieved.

enzyme inhibitors. Improvement inthe yield of prednisolone from hydro-cortisone by A. simplex usinginhibitors has been reponed recently.Apart from .1,1-dehydrogenase. A.simplex contains another enzyme. 20~-ketoreductase. which can reducehydrocortisone and prednisolone inthe culture. The use of iodoacelate andp-chloromercuric benzoic acid inhibit-ed the 20 j3-keloreductase withoutaffecting the .1,1-dehydrogenase activi-ty. The yield of prednisolone wasimproved with the addition ofinhibitors and selection of medium. Avery high degree of steroid transfor-mation (>98%) by Flavobacteriumdehydrogenans could be reached in atwo-liquid-phase system. The prcduc-tion of androst-a-ene-Lt't-dicne bythe microorganism from androsten-olone acetate in octane culture(vel/vel = 1:1) two-liquid-phase sys-tem was compared with the conver-sion rate in a Tween 80 containingmedium commonly used in industryand in a culture where no organic sol-vent or detergent was added. The rateof production of androst-4-ene-3.17-dione at the optimal cell density in thetwo-liquid-phase system and Tween80 containing medium was about 6and 1.5 times as high, respectively. asin an aqueous medium.

Future possibilitiesRemarkable progress already has beenmade on the applications of microbialsystems to steroid transformations.Further developmental activities relat-ing 10 controlled multiple transforma-

217

BIOTECHNOLOGY

Biotechnology meetings, literature update

lions for the purpose of cost reductionare expected. Encouraging results inordered sequential fennentations withvegetative cells already have beenobtained. There is the distinct possi-bility of further developments on con-ducting two or more microbial steps inone fermentation using mixed cul-tures. In mixed fermentation it isessential that the two microorganisms

To aid INFORM readers seeking 10keep up-to-date on the various aspectsof biotechnology of fats, oils andrelated materials, INFORM AssociateEditor James Rattray has compiled thefollowing listing. An earlier listappeared in the October 1990 issue ofINFORM.

Conferences, meetingsSecond Annual Conference-Fat and

Cholesterol Reduced Foods, March14-16.1991, Atlanta. Georgia.Information: IBC U.S.A. Confer-ences Inc., 8D Pleasant St., SouthNatick, MA 01760.

Bioexpo 91, April 9-12. 1991. Paris.France. Information: Bioexpo/Sepfi, 8 rue de la Mtcbodiere.75002 Paris, France.

American Chemical Society NationalMeeting, April 14-19, 1991,Atlanta, Georgia. Information:Department of Meetings and Divi-sional Activities. ACS, J 155·16thSI. NW, Washington. DC 20036.

Advances in Biotechnology 2: Biore-actors in Industry, April 14-25.1991. Edinburgh. Scotland. In for-mation: Courses Department. TheBritish Council, 65 Davies SI.,London WIY 2AA. United King-dom.

Expoflore International Exhibition ofPlant Biotechnology and New veri-eties of Plants, April 18-21, 1991,Geneva, Switzerland. Information:Expoflore. 8 rue du 31-Decembre,CH 1207 Geneva, Switzerland.

American Society for MicrobiologyAnnual meeting, May 5-9. 1991.Dallas, Texas. Information: MelissaSordyl, Meetings Department,ASM, 1325 Massachusetts Ave.

are capable of inducing the desiredenzymes in one another's presence.Development of new steroids of use-ful biological activities requiring newbiotechnology may receive greaterattention, Continuing attempts areexpected for utilization of sterol frac-tions obtained as industrial by-prod-ucts in the production of useful steroidintermediates. Greater emphasis on

the application of the newer conceptsof genetic engineering of microorgan-isms for their improvement as steroidtransforming agents or as agents forde 110\'0 steroid biosynthesis is antici-pated. In fact, as already mentioned. afew applications of transposon muta-genesis for improvement of microbialstrains already have been initiated.

NW. Washington. DC 20005,8th International Symposium on

Preparative Chromatography. May13-15. 1991, Arlington, Virginia.Information: Janet Cunningham.Barr Enterprises, PO Box 279.Walkersville. MD 21793.

Anachema 91. June 9-15. 1991,Frankfurt. Germany. Information:Decheme. Postfach 97 01 46. D-6000 Frankfurt am Main 97. Ger-many.

5th Annual Seminar on AnalyticalBiotechnology, June 10-13. 1991,San Francisco. California. Informa-tion: Janet Cunningham. BarrEnterprises. PO Box 279, Walk-ersville. MD 21793.

1991 World Congress on Celt and Tis-sue Culture. June 16-21, 1991,Anaheim. California. lnformution:1991 World Congress on Cell andTissue Culture. PO Box 73230,Baltimore, MD 21273.

Molecular Biology Biotechnology: ALaboratory-Based Introduction,July 22-26, 1991. Hatfield. Eng-land. Information: L. Hart. RoyalSociety of Chemistry, BurlingtonHouse, Piccadilly. London WI VOBN. United Kingdom.

BCCB First Conference and Exhibi-tion, Sept. 24-27, 1991, Leeds.England. Information: BiotechU.K .. c/o SCI. 14 Belgrave Square,London. WCIX 8PS, United King-dom.

8th World Congress of Food Scienceand Technology. Sept. 29-Oc1. 4,1991, Toronto, Canada. Informa-tion: Jean Allen, 3340 OrlandoDrive. Mlsstssauga. ON. CanadaLAV IC?

Third International Congress of Plant

Molecular Biology, OCI. 6-12,1991, Tucson, Arizona. Informa-tion: ISPMB. Woo Wester Confer-ence, Consultants, 2934 1(2. Bever-Iy Glen Circle, Suite 383. LosAngeles. CA 90077.

Advances in Genetic Technology:Feeding the World, Jan. 17-24,1992, Miami Beach, Florida. lnfor-rnaucn. The 1992 Miami Bio/Tech-nology Winter Symposium, PO Box016129, Miami. FL 33101-6129.

BooksAgricultural Biotechnology: Opponu-

nities for tntemationat Develop-ment. edited by G.J. Persley,C.A.B. International, Wallingford,Oxcn.. England, and Tucson, Ari-zona, 1990.

Rapeseed-s-Chemistry and Technolo-gy. H. Niewiadomski, Elsevier,Amsterdam, The Netherlands andNew York, 1990.

Plant Gene Transfer. edited by C.J.Lomb and R.N, Beachy, Wiley-Liss, New York, 1990.

Plant Cell and Tissue Culture, editedby J.w. Pollard and J.M. Walker,Humana, Clifton. New Jersey,1990.

Herbicides and Plant Metabolism,edited by A.D. Dodge. CambridgeUniversity Press. Cambridge. Eng-land, and New York, 1990.

Yeasts and Yeast-like Organisms, AKockova-Kratochvflova, VCH,Weinheim, Germany. and NewYork, 1990.

Yeast Technology, edited by J,F.T,Spencer and D.M. Spencer.Springer- Verlag. Berlin, Germanyand New York. 1990.

Biocatalysis. edited by D.A. Abramo-

INFORM. Vol. 2, no. 3 (March 1991)

218

BIOTECHNOLOGY

wicz, Van Nostrand Reinhold. NewYork. 1990.

Enzymes in Industry, edited by W.Gerhartz, VCH, Weinheim. Ger-many. and New York. J 990.

New journalsAgBiouch News and Information.

C.A.B. International, wallingford,Onon .• England. and New York,1989.

Bio-Organic & Medicinal Chemistryteners, Pergamon, Oxford, Eng-land. and Elmsford, New York.1990.

Btoseparauon, Kluwver, Dordrecht,The Netherlands, and Hingham,Massachuseus, 1990.

Trends in Food Science and Technolo-gy. Elsevier, Barking. Essex, Eng-land,199O.

Journal articlesBrowse. J .• B. Lemieux. D. Guerra

and C. Somerville. UCLA Symp.Mol. Cell. Bioi., New Ser. 129:301(1990). Strategies for ModifyingPlant Lipid Composition.

Branch, W.O., T. Nakayama and M.S.Chinnan, J. Am. Oil Chern. Soc.67:591 (1990), Fatty Acid Varia-tion among U.S. Runner-typePeanut Cunivars.

Sperling, P.. U. Hammer. W. Friedtand E. Heinz, Z. Naturforscn., C:Biosci. 45:166 (1990), High-oleicSunflower: Studies on Composi-tion and Desaturation of AcylGroups in Different Lipids andOrgans.

Yang. C. Y.S., and A.C. Peng, 1. Am.Oil Chern. Soc. 67:594 (1990),Analysis of Soybean Oil fromOhio.

Bhatty, R.S., and G.G. Rowland, 1.Am. Oil Chem. Soc. 67:364 (1990),Measurement of a-Linolenic Acidin the Development of Edible OilFlax.

Plelnes. S., and W. Friedl. Theor.Appl. Genet. 78:793 (1989), Genet-ic Control of Linolenic Acid Con-cemration in Seed Oil of Rapeseed(Brassica napus L.).

Cbrtsrou. P., D.E. McCabe. BJ. Mar-tinel l and W.F. Swain, TrendsBiotechnol, 8:145 (1990). SoybeanGenetic Engineering--CommercialProduction of Transgenic Plants.

Fehling. E .. and K.D. Mukherjee,Phytochemistry 29: 1525 (1990),Biosynthesis of TriacylglycerolsContaining Very-Iong-chain Mono-unsaturated Fatty Acids in Seeds ofLunaria annULI.

Bernerth, R., and M. Frenuen. PlantSci. 67:21 (1990). Utilization ofEurocyl-CoA by Acyltransferasesfrom Developing Seeds of Brassi-co napus (L.) Involved in Triacyl-glycerol Biosynthesis.

Saxena, P.K.. D. Williams and J. King,Plant Sci. 69:231 (1990). TheSelection of Chlorsulfuron-resis-rent Cell Lines of Independent Ori-gin from an Embryogenic Cell Sus-pension Culture of Brassica nopusL.

Cho, S.Y., 'Lw, Kwon and S.H. Yoon,1. Am. Oil Chern. Soc. 67:558(1990), Selective Removal of FreeParty Acids in Oils Using a Micro-organism.

Jeme]c. K., A. Cimerman. M. Ven-dramin and A. Perdih , Appl.Microbial. Btotechnol. 32:699(1990), Lipids of Citric Acid-pro-ducing Aspergillus niger StrainGrown in Copper- and in Man-ganese-supplemented Media.

AI-Hasan. R.H .. A.M. Ali and S.S.Radwan, Mar. Bioi. 105:9 (1990),Lipids, and Their Constituent FattyAcids. of Phaeocystis from theArabian Gulf.

Parks, W.S .. P.A. Murphy and B.A.Glatz, Can. 1. Microbial. 36:318(1990). Lipid Metabolism and CellComposition of the OleaginousYeast Apiotrichum curvatumGrown at Different Carbon toNitrogen Ratios.

Turcotte, G .• and N. Kosaric, Adv.Biochem. Eng.lBiotechnol. 40:13(1989). Lipid Biosynthesis inOleaginous Yeasts.

Davies. RJ .• J.E. Holdsworth and S.L.Reader. Appl. Microbial. Biotech-not. 33:569 (1990). The Effect ofLow Oxygen Uptake Rate on theFatly Acid Profile of the Oleagi-nous Yeast Apiotrichum cllnatum.

Singh, B.. G.K. Oberoi and S.c. Shar-ma, Indian 1. Exp, Bioi. 28:430(1990), Effect of pH Stress onLipid Composition of Saccha-romyces cerevisiae,

Tsuzuki, M., E. Ohnuma. N. Saw. T.

Takaku and A. Kawaguchi. PlantPhysiol. 93:851 (1990), Effects ofCarbon Dioxide Concentration dur-ing Growth on Fatty Acid Compo-sition in Microalgae.

Dedyukhina. E.G .• R.1. Nugumanova.O.A. Reshetnik and V.K. Eroshin,Prikl. Biokhim. Mjkrobiol. 26:67(1990); English translation: Appl.Biochem. Microbial. 26:52 (1990),Effect of the Water-soluble Antiox-idant MannichBase Acetate on meTrichosporon pullulans Growthand Lipid Synthesis (-single celloil production).

Hirano. M .. H. Mori, Y. Miura. N.Matsunaga, N. Nakamura and T.Matsunaga. Appl. Biochem.Biotechnol. 24-25:183 (1990). "(-Linolenic Acid Production byMicroalgae.

Sajbidor. J .. S. Dobrodova and M.Certfk, Biotechnol, Leu. 12:455(1990). Arachidonic Acid Produc-lion by Mortierella sp. 5-17. Influ-ence of C/N Ratio.

Rezanka, T.. M. Yu. Sokolov and I.Viden, FEMS Microbiol. Ecol.73:231 (1990), Unusual and Very-long-chain Fatty Acids in Desulfo-tomacutum, a Sulfate-reducingBacterium.

Sriharan, S.• D. Bagga and T.P. Sriha-ran, Appl. Biochem, Biotechnol,24-25:309 (1990), Effects of Nutri-ents and Temperature on Lipid andFatly Acid Production in theDiatom Honuschia DI-60.

Huehne. K .• M. Schweizer and E.Schweizer. Fat Sci. Technol.92:232 (1990). Genetic Manipula-tion of Fatty Acid Chain LengthPattern in Yeast.

Spitzer, V., F. Marx. J.G.S. Maia andK. Pfeitsticker. Fette Wiss. Techno!.92:165 (1990). Curupira tefeensis(Olacaceae)-a Rich Source ofVery Long Chain Fatty Acids.

Carlson. K.D., A. Chaudhry, R.E.Peterson and M.O. Bagby, 1. Am.Oil Chern. Soc. 67:495 (1990).Preparative Chromatographic Iso-teuon of Hydroxy Acids from us-cueretta [endleri and L gordoniiSeed Oils.

Harris, P.L.. S.L. Cuppett and L.B.Bullerman. 1. Food Prot. 53:176(1990), A Technique Comparisonof Isolation of Lipolytic Bacteria.

INFORM, Vol. 2, no. 3 (March 1991)

r------------- - -- ----

219

John. B.N., J.D. Alurralde and J.Klein, App/. Microbial. Biotechnoi.33:367 (1990), Lipase Productionby Free and Immobilized Proto-ptasts of Sporotrichum (Cnryso-sporium) thermophile Apinis.

Kamaly, L.M., K. Takayama and E. H.Marth, J. Dairy Sci. 73:280 (1990),AcyJglyceroJ AcylhydroJase(Lipase) Activities of Streptococ-cus lactis, Streptococcus cremonsand Their Mutants.

Feller, G .. M. Thiry, J.L. Arpigny, M.Mergeay and C. Gerday, FEMSMicrobial. Lett. 66:239 (1990),Lipases from Psychrophilic Antarc-tic Bacteria.

lizumi, T.. K. Nakamura and T.Fukase, Agrie. Bioi. Chern.54:1253 (1990), Purification andCharacterization of a ThermostableLipase from Newly Isolated Pseu-domonas sp. KWI-56.

Vicente, M.L.e .. M.R. Aires-Barrosand J.M.S. Cabral, Biotechnol.Tech. 4:137 (1990), Purification ofChromobacterium viscosum Lipas-es Using Reversed Micelles.

Vogel, R.F., W. Gaier and W.P.Hammes, F£MS Microbiol. Leu,69:289 (1990), Expression of theLipase Gene from Staphylococcushyicus in Lactobacillus curvatusKc2-c.

Arnold, F.H .• Trends Biotechnol.8:244 (1990), EngineeringEnzymes for Nonaqueous Solvents.

Chekrabant, S., S. Chanda and S.Matai, Indian 1. Microbial.30:75-77 (1990), Influence of Cand N Sources on Lipase andBiomass Production by a SoilStrepsomycete.

Alhir, S.• P. Markakis, and R.C. Chan-dan, 1. Agric. Food Chem. 38:598(1990), Lipase or Penicillium case-icalum,

Nakashima, T., S. Kyctani, E. Izumo-to and H. Fukuda, 1. Ferment. Bio-engin. 70:85 (1990). Cell Aggrega-tion as a Trigger for Enhancementof Intracellular Lipase Productionby a Rhizopus Species.

veeraragavan. K.• T. Colpitts and B.F.Gibbs, Biochim, Biophys. Acta L1044:26 (1990), Purification andCharacterization of Two DistinctLipases from Geotrichum can-didum,

BIOTECHNOLOGY

(1990), Enzyme Catalysis inMicroemulsions: Enzyme Reuseand Product Recovery.

Holmberg, E., P. Szmulik, T. Norinand K. Huh. Biocotalysis 2 :217(1989). Hydrolysis and Esterifica-tion with Lipase from Candidacytindracea. Influence of the Reac-tion Conditions and Acid Moietyon the Enantiomeric Excess.

Kery. V.• J. Haplova, K. Tihlarik andS. Schmidt, J. Chem. Technol.Biotechnot, 48:201 (1990), FactorsInfluencing the Activity and Ther-mcstabiluy of Immobilized PorcinePancreatic Lipase (-immobiliza-ticn on cellulose bead support).

Loose, S .. D. Meusel. A. Muschrerand B. Ruthe, Nahrung 34:37(1990), Water Binding of Adsorp-live Immobilized Lipases.

lnada, Y.. A. Matsushima, K. Taka-hashi and Y. Saito, Biocatatysis3:317 (1990), Polyethylene Glycol-modified Lipase Soluble andActive in Organic Solvents.

Khurana, A.L., and C.T. Ho, 1. Liq.Chromatogr. 13:715 (1990), Immo-bilization of Porcine-pancreaticLipase on Silica Support To StudyLipolysis and Reverse HydrolysisReaction.

Hayashi, T.• and Y. lkada, Biotechnol.Bioengin. 36:593 (1990), Lipopro-tein Lipase Immobilization ontoPolyacrolein Microspheres.

Wood. L.L .. C.S. Cobbs. L. Lantz II.L. Peng and G.J. Calton, 1.Biotechnol. 13:305 (1990), Immo-bilization of Enzymes withPolyaziridines (-lipase).

Kosugt, Y., H. Tanaka and N. Tomizu-ka, Biotechnol, Biaengin, 36:617(1990), Continuous Hydrolysis ofOil by Immobilized Lipase in aCountercurrent Reactor.

Iso, M.. H. Matsumoto, S. Urushiya-rna and S. Omi. 1. Microencopsu-toston 7: 167 (1990), Application ofEncapsulated Enzyme Dispersed ina Continuous Stirred Tank-Reactor(-lipase).

van der Padt. A., M.J. Edema, J.J.W.Sewalt and K. van't Riet, 1. Am.Oil Chem. Soc. 67 ..347 (1990).Enzymatic Acylglycerol Synthesisin a Membrane Bicreactor;

Bloomer, S., P. Adlercreutz and 8.Mattiasson. 1. Am. Oil Chem. Soc.

Shimada, Y.. A. Sugihara, T. lizumiand Y. Tominaga, 1. Biochem,107:703 (1990). cDNA Cloningand Characterization of Geotri-chum candidum.

Sanz, L.C.. and J.M. Olias, FoodChem. 37:221 (1990), Characteri-zation of Lupine Seed Lipase.

Rae, K.V.S.A., M.M. Paulose and G.Lakshminarayana, Biotechnol. Lett.12:377 (1990), In Situ Lipolysis ofCastor Oil in Homogenized CastorSeeds.

Jensen, R.G., O.R. Galluzzo and V.J.Bush; Biocatalysis 3:307 (1990),Selectivity is an Important Charac-teristic of Liposes (Acyl-glycerolHydrolases).

Kannappan, V., Anal. Biochem.186:301 (1990), A Simple andSensitive Method for the Estima-tion of Microbial Lipase Activity.

Hochkoeppler. A .. and S. Palmier,Biocatalysis 3:357 (1990). A Spec-trophotometric Lipase ActivityAssay by Coupled Lipase-Lipoxy-genase in Reverse Micelles.

Wu. S.H., Z.W. Guo and C.J. Sih,l.Am. Chern. Soc. 112:1990 (1990),Enhancing the Enantio Selectivityof Candida Lipase Catalyzed EsterHydrolysis via NoncovalentEnzyme Modification.

Hoshino, T .• T. Yamane and S.Shimizu, Agrie. Bioi. Chem.54:1459 (1990). Selective Hydrol-ysis of Fish Oil by Lipase To Con-centrate n-3 Polyunsaturated FattyAcids.

Hills, M.J., I. Kiewitt and K.D.Mukherjee, 1. Am. Oil Chern. Soc.67:561 (1990), Enzymatic Frac-tionation of Fatty Acids: Enrich-ment of v-Ltnotenlc Acid andDocosahexaenoic Acid by Selec-tive Esterification Catalyzed byLipases.

Halling, P.J.. Fat Sci. Technol, 92:74(1990), Lipase-catalyzed Modifica-tion of Oils and Fats in OrganicTwo-phase Systems.

Ferreira, G .. and J.S. Patton. 1. LipidRes. 31:889 (1990), Inhibition ofLipolysis by Hydrocarbons andFatty Alcohols (-response of pan-creatic lipase),

Larsson, K.M., P. Adlercreutz, B.Mattiasson and W. Olsson,Bioteclmol, Bioengin. 36:135

1NFORM, Vol. 2, no. 3 (March 1991)

220

BIOTECHNOLOGY

67:519 (1990), Triglyceride Inter-esterification by Lipases. I. CocoaButter Equivalents from a Fractionof Palm Oil.

Holmberg. E., and K. Hult, Biocataiy-sis 3:243 (1990). Transesreriflca-tion with Candida cylindraceaLipase in II Biphasic Aqueous-organic System. Dependence of theEnantiomeric Ratio and the Reac-tion Rate on the Proportions ofwerer and Cyclohexane.

Hirata. H .. K. Higuchi and T.Yamnshina, J. Biatechnol, 14:157(1990). Lipase-catalyzed Transes-terlflcanon in Organic Solvent:Effects of Water and Solvent, Ther-mal Stability and Some Applica-tions.

Goldberg, M., D. Thomas and M.D.Legoy, £ur. 1. 8iochem. 190:603(1990). The Conuol of Lipase-cal-alyzed Transesterif'icarion andEsterification Reaction Rates.Effects of Substrate Polarity. WaterActivity and Water Molecules onEnzymatic Activity.

Boyer, 1.L., B. Gilot and R. Guiraud,Appl. Biochem. Biotechnoi. 33:372(1990). Heterogeneous EnzymaticEsterification: Analysis of theEffect of Water (-fatty acid.ferryalcohol esterification).

Boyer. 1.L .. B. Gilot and R. Guiraud,Appl. Biochem, Biotechnol.24-25:201 (1990). Esterificationwith Solid Enzymes: Water andSolvent Influence (-studies onMucor miehei lipase).

Rosuup-Nielscn. T .. L.S. Pedersenand J. Villadsen. J. Ctiem. Technol.Biotecbnot, 48:467 (1990), Ther-modynamics and Kinetics ofLipase Catalyzed Hydrolysis ofOleyl Oleate.

Ota. T.. S. Takano and T. Hasegawa.Agric. Bioi. Chem. 54:1571 (1990).Synthesis of C18'Fauy Acid Estersin Organic Solvent by Lipase fromCandida cylindracea.

Laumann. R.. O. Ghisalba, D. Gyga,H.P. Schaer and E. Schmidt, Bio-catalysis 3:137 (1990). Screeningand Applications of MicrobialEstcrases for the EnantioselectiveSynthesis of Chiral GlycerolDerivatives (-use of Mucorjavanicus lipase).

Liu. K.C., K. Nezaki and C.H. Wong.

Biocatalysis 3:169 (1990). Prob-lems of Acyl Migration in Lipase-catalyzed Enantioselective Trans-formation of rneso-I, 3-diol Sys-tems.

Knez. Z.•M. Leitgeb, D. Zavrsnik andD. Lavric , Fette Wiss. Technol.92:169 (1990), Synthesis of OleicAcid Esters with ImmobilizedLipase.

Ramaswamy. S.• B. Morgan and A.C.Oehlschlager, Tetrahedron Lett.31 :3405 (1990), Porcine PancreaticLipase Mediated Selective Acyta-tion of Primary Alcohols in Organ-ic Solvents.

Langrand, G .. N. Rondot, C. Trianta-phylides and 1. Baratti, Biatechnot,Lett. 12:581 (1990), Short ChainFlavour Ester Synthesis by Micro-bial Lipases.

Feichter. C.. K. Faber and H. Griengl.Biocatalysis 3:145 (1990). Cherne-enzymatic Preparation of OpticallyActive Long-Chain 3-Hydroxy-al kanoares (-use of Candidacylindracea lipase).

Ducret. A .. M. Pina, D. Ornnrer and J.Graille. Oteagineux 44:603 (1989),Enzymic Deacidification of Hyper-acid Oils.

Svensson. I.. P. Aldercreutz and B.Mantasson, Appl. Microbial.Biotechnol, 33:255 (1990). Inter-esterification of Phosphat idyl-choline with Lipases in OrganicMedia.

Hirashima. Y., A.A. Farooqui. E.J.Murphy and L.A. Horrocks, Lipids25:344 (1990). Purification ofPlasmalogens Using Rhi zopusdelemar Lipase and Naja naja nojaPhospholipase A2. •

Sada, E.. S. Katoh and A. Kheirclo-moom, Appl. Biochem. Biotechnol.22:247 (1989), Effects of Arachi-donic Acid Concentration onProstaglandin Biosynthesis andFeasibility of Semi batch Process.

Babiak, K.S., 1.S. Ng. 1.H. Dygos.C'L. Weyker. Y.F. Wang and C.H.Wong. J. Org. Chem. 55:3317(1990), Lipase-catalyzed lrre-versible Transesterification UsingEnol Esters: Resolution ofProstaglandin Synthons 4-Hydrox s- 2-al k y 1-2-c yctopen-tencnes and Inversion of the 4SEnantiomer to the 4R Enantiomer.

Ghosh. D .• E. Mukherjee and 1. Duna,Indian 1. Biachem, Biophys. 27:7(1990). Stabilization of Prosta-glandin-synthetase by Immcbiliza-tion of Goat Seminal Microsomeson Silica Gel-G.

Kikuchi. S., and E. Kolauukudy,Agric. BioI. Chern. 54:1411 (1990).Immobilization of Fatly Acid Syn-thetase from Mycobacterium smeg-matis by Radiation-induced Poly-merizmion.

Arnezder. C., and W.A. Hampel.Biotechnol, Lett. 12:277 (1990),Influence of Growth Rate on theAccumulation of Ergosterol inYeast Cells.

Sz ykula 1.• C. Hebda and 1.Orptszewskt. Biotechnol, Lett.12:505 (1990). Isolation and Iden-tification of a New Metabolite ofMicrobial Conversion of UpgradedNeutral Fraction of Polish Tall Oilby Means of Four Strains ofMycobacterium sp. (-5a-andro-stane-3,6,17-trione).

Ramelmeier, R.A., and H.W. Blanch.Btocatatysis 2 :97 (1989). MassTransfer and Cholesterol-oxidaseKinetics in a Liquid-liquid Two-phase System (-use of Strepto-myces sp. enzyme).

Johnson. T.L .• and G.A. Somkuti, 1.Food Prot. 53:332 (1990). Proper.ties of Cholesterol Dissimilation byRhodococcus equi (-treatment ofegg yolk preparations).

Lee, K.M., and J.F. Biellmann,Biochimie 72:285 (1990). 13-Hydroxysteroid-dehydrogenase:Activity in Microemulsion andExtraction from Pseudomonastestosteroni Cells with Microemul-slcn.

Seidel. S.. W. Kreis and E. Reinhard.Plant Cell Rep. 8: 62 (1990). llj·3p-Hydroxysteroid Dehydroge-nase/llj-ll"-Ketosteroid Isomerase(3P-HSD), a Possible Enzyme ofCardiac Glycoside Biosynthesis inCell Cultures and Plants of Digi-talis lanata EHRH.

Prabha, V., M. Gupta and K.G. Gupta.Can. J. Microbiol. 35:1076 (1989).Kinetic Properties of 7a-Hydroxy-steroid Dehydrogenase fromEscherichia coli 080 (-bile acidoxidation).

Hudnik-Plevnik, T., and B. Cresnar, J.

INFORM. Vol. 2. 00. 3 (March 1991)

221

BIOTECHNOLOGY

Steroid Biochem. 35:749 (1990),Factors Affecting the Induction ofI l c-Hydroxylese of Progesteronein the Filamentous Fungus Rhizo-pus nigricans.

Zakelj-Mavric, M., A. Plemenitas, R.Kamel and I. Belie, J. SteroidBiochem. 35:627 (1990), 1113-Hydroxylation of Steroids byCochliobolus Iunatus.

Chen, K-C., and H-C. WeYt EnzymeMicrob. Technol. 12:616 (1990),Dissolution-enzyme Kinetics oflip-Hydroxylation of Conexoloneby Curvularia lunata.

Paper. D.H., and G. Franz, Plant CellReports 8:651 (1990), Biotransfor-mation of 5p-H-Pregnan-3p-ol-20-one and Cardenolides in Cell Sus-pension Cultures of Nerium olean-derL.

Kawaguchi, K., M. Hirotani, T.Yoshikawa and T. Furuya, Phyto-chemistry 29:837 (1990), Biotrans-formation of Digitoxigenin by Gin-seng Hairy Root Cultures.

Greidziak, N., and B. Diettrich, Plan-fa Med. 56:175 (1990), Balch Cul-tures of Somatic Embryos of Digi-talis tanata in Gas-lift Perrnenrors,Development and CardenolideAccumulation.

Saito, K., M. Yamazaki, K. Shimomu-fa. K. Yoshimatsu and I. Mura-koshi, Plant Cell Reports 9:121(1990), Genetic Transformation ofFoxglove (Digirafis purpnrea) byChimeric Foreign Genes and Pro-duction of Cardioactive Glycosides(-saponin production).

Ward. O.P., and e.S. Young, EnzymeMicrob. Technol. 12:482 (1990),Reductive Transformations ofOrganic Compounds by Cells orEnzymes of YeaSL

Kise, S., and M. Hayashida. J.Biotechnol. 14.'221 (1990). Two-phase System Membrane Reactorwith Cofactor Recycling.

Ouotine. G., G. Carrea, S. Riva andA.F. Buckmann, Enzyme Mierob.Technol. 12:596 (1990). Coenzy-matic Properties of Low Molecu-lar-weight and Macromolecular N6_Derivatives and NAO' and NADP"with Dehydrogenases of Interestfor Organic Synthesis.

Graff, G., L.A. Anderson and i.w.Jaques, Anal. Biochem. 188:38

(1990), Preparation and Purifica-tion of Soybean Lipcxygenase-derived Unsaturated Hydroperoxyand Hydroxy Fatty Acids andDetennination of Molar Absorptiv-ides of Hydroxy Fatty Acids.

Apre. S.S., N. Weber and H.K. Man-gold, FEBS Lerr. 265:104 (1990),Biologically Active Ether Lipids:Biotransformation of rac-I(3)-0-Alkylglycerols in Cell SuspensionCultures of Rape and Semisynthe-sis of I-O-Alkyl-2-palmitoyl-sn-g Iycero- 3 -phospho-(N -palmi toy I)ethanolamines, Potent Anti-tumorAgents.

Berger, R.G., Z. Akkan and F. Draw-en, Z. Naturforsch. C 45:187(1990), The Essential Oil ofColeonema album (Rutaceae) andof a Photomixotrophic Cell CultureDerived Thereof.

Gbolade, A.A., and G.B. Lockwood,J. Plant Physiol. 136:198 (1990),Metabolic Studies of Volatile Con-stituents in Tissue Cultures of Pet-roselinum crispum (Mill) Nyman.

Spencer, A., J.D. Hamill and M.J.C.Rhodes, Plant Cell Reports 8:601(1990), Production of Terpenes byDifferentiated Shoot Cultures ofMentha citrata Transformed withAgrobacterium tumefaciens T37(-products for flavor industry).

Toyota, M., and K. Hcsteumenn, Phy-tochemistry 29.'1485 (1990), Anti-fungal Diterpenic Esters from theMushroom Boletinus cavipes.

Banthorpe, D.Y., J.T. Brown and G.S.Morris, Phytochemistry 29.'2145(1990), Accumulation of the Anti-fungal Diterpene Selareol by CellCultures of Salvia sctarea andNicotiana glutinosa,

Gbolade, A.A., and G.B. Lockwood,Z. Naturforsch. C 45:245 (1990),Biotransformation of Monoter-penes by Polyurethane FoamImmobilized Cells of Petroselinumcrtspom (Mill) Nyman.

Hook, I., R. Lecky, B. McKenna andH. Sheridan, Phytochemistry29:2143 (1990), Biotransfonnationof Linalyl Acetate by SuspensionCultures of Papaver bracteauon.

Takaichi. $., K. Shimada and J-1.Ishidsu, Arch. Microbial. 153:118(1990), Carotenoids from the Aero-bic Photosynthetic Bacterium Ery-

throbacter longus.' (3-Carotene andIts Hydroxyl Derivatives.

An, Gj-l., and E.A. Johnson, AnionicLeeuwenhoek 1. Microbial. 57.'191(1990), Influence of Light onGrowth and Pigmentation of theYeast Pha/fia rhodozyma (-car-otenoid production).

Lers, A .• Y. Biener and A. Zamir,Plant Physial. 93.'389 (1990), Pho-toinduction of Massive I3-CaroteneAccumulation by the Alga Ouna-liella bardawil.

Ben-Amou, A., and M. Avron, TrendsBiotechnol, 8.'121 (1990), TheBiotechnology of Cultivating theHalotoleram Alga Dunalielta (-13-carotene, glycerol production).

Lardet, L., F. Enjalric and M.P. Car-ron, C.R. Seances Acad. Sci. Ser.3310: 195 (1990), Hevea brasilien-sis (Mull. Arg.) Apex Cultures.Influence of the MorphologicalStage and Explant Age (-rubbershoot culture).

Takeuchi, K., K. Koike and S. Ito, J.Biotechnol. /4:179 (1990), Produc-tion of cis-Unsaturated Hydrocar-bons by a Strain of Rhodococcus inRepeated Batch Culture with aPhase Inversion. Hollow Fiber Sys-tem.

Morgan, P., and R.J. Watkinson.Water Sci. Technol, 22:63 (1990).Assessment of the Potential for tnSitu Biotreatment of Hydrocarbon-contaminated Soils.

Sorkhoh, N.A., M.A. Ghannoum, A.S.Ibrahim, R.J. Streuon and S.S.Radwan, Environ. Pollut. 65.'1(1990), Crude Oil and Hydrocar-bon-degrading Strains of Rhado-coccus rhodochrous Isolated fromSoil and Marine Environments inKuwait.

Shafeeq, M., D. Kolub, Z.M. Khalid,A.M. Khan and K. Malik. 1.Appl.Microbial. Biotechnol. 5:505(1990), Degradation of DifferentHydrocarbons and Production ofBiosurfactant by Pseudomonasaeruglnosa Isolated from CoastalWater.

Bryant, F. 0 .. App/. Environ. Microbi-01. 56:1494 (1990), ImprovedMethod for the Isolation of Blosur-factant Glycolipids from Rhoda-COCCIIS sp. Strain H 13A.

Matsuyama, T.. K. Kaneda, I. Ishizu-

INFORM, Vol. 2, no. 3 (March 1991)

222

BIOTECHNOLOGY

ka. T. Toida and I. Yano, J. Bacte-rial. 172:3015 (1990), Surface-active Novel Glycolipid andLinked 3-Hydroxy Fatly AcidsProduced by Serratia rubidaea.

Kim, 1.S.• M. Powa.lla, S. Lang, F.Wagner, H. luensdorf and V. Wray,Folia Microbial. 34:398 (1989).Microbial Glycolipid Productionunder Nitrogen Limitation andResting Cell Conditions. .

Kondrat'eva, E.N., and E.N. Krasil'nikova, Prikl, Biokhim, Mikrobiol.25:785 (1989); English translation:APIJI. Biochem. Microbial. 25: 665(1990), Phototrophic Bacteria asProducers of Pcly-jf-Hydroxybu-tyrate.

Ramsay, B.A., K. Lomaliza, C.Chavarie, B. Dube, P. Bataille andJ.A. Ramsay, Appl. Environ.Microbial. 56:2093 (1990). Pro-duction of Poly(-Il-hydroxybulyric-coop-hydroxy valerie) Acids(-bacterial production).

Timm, A., D. Byrom and A. Stein-bilchel, Appl. Microbiol. Biotech-nolo 33:296 (1990), Formation ofBlends of Various Poly(3-hydrox-yalkanoic acids) by a RecombinantStrain of Pseudomonas aleovarans.

Ramsay, J.A., E. Berger. B.A. Ramsayand C. Chavarie, Biotechnol. Tech.4: 221 (1990), Recovery of Poly-3-Hydroxyalkanoic Acid Granules bya Surfactant-Hypochlorite Treat-ment (-production by Alcaligeneseutrophus}:

Doi, Y.. A. Segawa and M. Kunioka,1111.1. Bioi. Macromol. 12:106(1990), Biosynthesis and Charac-terization of Poly(3-hydroxy-butyrate-co-4-hydroxybutyrate) inAlcaligenes eutrophus.

Pries. A .. A. Steinbiichel and H.G.Schlegel, Appl. Microbiol. Biotech-nol. 33:410 (1990). Lactose- andGalactose-utilizing Strains of Poly(hydroxyalkanoic aeid)-accumulat-ing Alcaligenes eutrophus andPseudomonas saccharophila.Obtained by Recombinant DNATechnology.

Tal. S .. P. Smimoff and Y. Okon. 1.Gen. Microbiol. 136:1191 (1990),The Regulation of Pcly-B-Hydrox-ybutyrate Metabolism in Azospiril-tum brasilense during BalancedGrowth and Starvation.

Bitar, A., and S. Underhill, Biotech-nol. Lett. 12:563 (1990). Effect ofAmmonium Supplementation onProduction of Pcty-BiHydroxybu-tyrate Acid by Alcaligenes ellfro-phus in Batch Culture.

Garcia Lillo, J .. and F. Rodriguez-Valera, Appl. Environ. Microbiol.56:2517 (1990), Effects of CultureConditions on Poly (~-hydroxybu-tyric acid) Production by Halofer-ax mediterranei.

Fritzsche, K., R. W. Lenz and R.C.Fuller, 1111.1. Bioi. Macromol.12:85 (1990), Production of Unsat-urated Polyesters by Pseudomonasoleovorans.

Ziokarnik, M .• Chem. Eng. Prog.86:62 (1990), Trends and Needs inBioprocess Engineering.

Frommer. w., Food Biotechnal, 4:613(1990), Safety Aspects in Biotech-nology.

Umehara, K.. Y. Masago. T. Mukaiya-rna and O. Okumura. Yukagakll39:32 (1990), Behavior of AlkalineLipase on Detergency (-inEnglish).

PatentsInstitute of Microbiological Produc-

tion (L. Romanova, M.V. Zalashkoand I.N. Stigailo), U.S.S.R. Patent1,541,249 (1990), Lipomycesstarkeyi as Lipid Source.

Suntory (K. Akirnotc. Y. Shinmen, H.Yamada and A. Shimizu). JapanPatent 1,304,892 (1989). MicrobialManufacture of PolyunsaturatedFatty Acid-enriched Fat or Oil.

Idemitsu Petrochemical (T. Shin-auchi), Japan Patent 1,300,897(1989), Manufacture of .t.\6.9.u.u_Ocradecaterraenoic Acid-contain-ing Lipids with Bacteria.

Suntory (K. Akimoto. Y. Shinmen, H.Yamada and S. Shimizu), EuropePatent 355,972 (1990). Process forProduction of Fatty Acids HavingHigh Degree of Unsaturation(-formation of j-linclenic acidand arachidonic acid by Echi-nosporangium traversalisy.

Nippon Oils and Fats (M. Kaji and T.Funada), Japan Patent 2.086,789(1990), Manufacture of Arachidon-ic Acid-containing Fats and Oilswith Emomophthoro Species.

Tosch Corporation: Sagami Chemical

(A. Oda, G. Uematsu. K. Yazawa.K. wetabe and S. Kondo), JapanPatent 2,145,191 (1990), Separa-tions of Highly Unsaturated FattyAcids from Marine Microorgan-isms.

Agency of Industrial Sciences andTechnology (Y. lkushima, I. Sate,K. Harada, S. Ito. T. Asano and T.Goto), Japan Patent 2,008,298(1990). Btcosapenraenotc Acid andDocosahexaenoic Acid Isolationand Purification from the Visceraof Cuttlefish.

Nippon Oils and Fats (H. Okuyama),Japan Patent 2,113.850 (1990).Anticancer Fat and Oil Composi-tions Containing (X-Linolenic Acid.

Sagami Chemical (K. Yazawa, K.Araki, C. Ishikawa and S. Kondo),Japan Patent 2,097.393 (1990).Manufacture of EicosapentaenoicAcid-containing PhospholipidCompositions by Marine Microor-ganisms.

Idemitsu Petrochemical (M. Sugano,H. Kikuji. K. Amano and T. Sano).Japan Patent 2.053,726 (1990),Bishomo-y-Linolenic Acid-con-taining Phospholipids and Anti-cholesteremics containing thePhospholipids and/or j-LtnotenfcAcid-containing Phospholipids(-manufactured by Conidiabotusheterosporus).

Kanegafuchi Chemical (T. Nakajimaand H. Fukuda), Japan Patent2.049.582 (1990). Cultivation ofFilamentous Fungi and Bacteria ToProduce High Levels of Lipase.

United States Department of Agricul-ture (T.R. Berka and M.J. Haas).U.S.A. Patent 369.975 (1989).cDNA Cloning of the Gene for theTriglyceride Lipase of Rhizopusdelemar.

Showa Denko (M. Nishioka. K. Joko,M. Takama and R. Matsui). JapanPatent 2,092,281 (1990), LipaseManufacture with Candida for Usein Detergents.

Agency of Industrial Sciences andTechnology (Y. Kosugi, H. Tanakaand M. Shiraki), Japan Patent2,092.282(1990). Immobilizationof Lipase on Powdered Anion-exchange Resins.

Kao (H. Yokomichi. T. Yasumasu andK. Nakamura), Japan Patent

INFORM. Vol. 2. no. 3 (March 1991)

223

BIOTECHNOLOGY

1.262.795 (1989). Immobilizationof Lipid-hydrolyzing Enzymes onMacroporous Chelate Resins.

Agency of Industrial Sciences andTechnology (M. Hayashida and S.Kise), Japan Patent 2.092,290(1990), Immobilization of Enzymeon Hydrophobic Porous Carrier inPresence of Inorganic Salt andwater-soluble Polymer (-lipase).

Shiseido (T. Kaminuma and K. Tomi-ta), Japan Patent 1.273,579 (1989),Degradation of Lipids with Immo-bilized Enzyme and ApparatusThereof (~Iive oil lipolysis).

National Research Council of Canada(F. Ergan, M. Trani and G. Andre),Europe Patent 354,850 (1990),Preparation of Immobilized Lipas-es and Their Use in the Synthesisof Glycerides.

Amano Pharmaceutical (T. Mase),Japan Patent 2,086,787 (1990),Enzymic Hydrolysis of Fats andOils (-use of Candida cylin-dracea lipase).

Itoh Oil (c. Yamaguchi. A. Toyama,S. Asaoka and F. Ogata). JapanPatent 2.013,389 (1990). Hydroly-sis of Castor Oil by Lipase (c-cseof fungal and yeast enzymes).

Shokuhin Sangyo Bioreactor System(T. Yamada, K. Osaro, Y. Osadaand K. Tanaka), Japan Patent2,142.484 (1990). Manufacture ofTriglycerides with Low Water Con-tent with Immobilized Lipase.

Novo Industri (8. Huge-Jensen), Dan-ish Patent 156,584 (1989). Immo-bilized Mucor mienei Lipase-basedPreparations for the Transesterifi-cation of Fats.

Enztech (A. Zaks and A.T. Gross).WO (Patent Cooperation Treaty)90,004,033 (1990), Production ofMonoglyceride by EnzymaticTransestenfication.

Sapporo Breweries (T. Nishio. K.Achinami, T. Konno and M.Kamimura), Japan Patent1,252,293 (1989), Optically ActiveGlycerol Monoesters and TheirEnzymic Preparation.

Mitsubishi Kasei (Y. Ora and H.Machida). Japan Patent 2.660,591(1990), Manufacture of SucroseFatty Acid Esters with Lipase.

Sapporo Breweries (T. Nishio, T.Konno. M. Kamimura. K. Achina-

mi, Y. Terao and M. Murata). JapanPatent 1.309,694 (1989), Manufac-ture of Optically Active Alcoholsby Enzymic Transesterification.

Agency of Industrial Sciences andTechnology (H. Hirata, H. Higuchi.T. Yamashina and K. Ishikawa),Japan Patent 2,086,797 (1990).Manufacture of Optically Active 2-Alkanol Esters with Lipase.

Genzyme (A.E. Walts and E.M. Fox),U_S.A_ Patent 4.923.810 (1990). ALipase Fraction for Resolution ofGlycidyl Esters of High Enan-tiomeric Excess.

BASF (0. Urban, W. Ladner. A. Pauland A. Sanner). German Patent3.819.467 (1989). Immobilizationof Swine Pancreatic Lipase forResolution of Racemic Ester.

Showa Sangyo (H. Takahashi), JapanPatent 2,049,593 (1990), EnzymicManufacture of Lysolecithins.

Fuji Chemicals (M. Shima, T.Takeuchi and M. Yuya), JapanPatent 2,027,996 (1990). Produc-tion of Optically Active Cyclopen-tenol Derivatives (-use of lipasefor prostaglandin precursor prepa-ration).

Iowa State University (D.C. Beitz,J.W. Young and S.S. Dehal),U.S.A. Patent 4.921.710 (1990).Plant Cholesterol Reductases forLowering Cholesterol Content ofFoods of Animal Origin.

Johns Hopkins University (D.W.Payne and P. Talalay). U.S.A.Patent 4,874,696 (1989), A NovelAlcaligenes Producing Steroid-transfonning Enzymes and Purifi-cation of the Enzymes.

Agency of Industrial Sciences andTechnology (Y. Kosugi. H. Tanaka,N. Tomizuka, K. Akeboshi, Y. Mat-sufune and S. Yoshikawa), JapanPatent 1,218.593 (1989). EnzymicManufacture of Sterol Fatty AcidEster.

Agency of Industrial Sciences andTechnology (M. Hayashida and S.Kise), Japan Patent 2,092.283(1990), Improving the Efficiencyof Immobilization of Enzymes onPorous Hydrophobic Carriers(-application to 3a-hydroxydehy-drogenase).

Snow Brand (M. lida. M. Yoshihamaand N. Miyala). Japan Patent

2,060,584 (1990), f3,-Hydroxy-steroid Oxidase and Its Manufac-ture with Rhodocaccus equi andCharacterization.

Snow Brand (M- Nakakoshi, Y. Tamu-ra. M. Yoshihama and N. Miyata).Japan Patent 2,101,096 (1990),Aromatase Inhibiting 7a, IIa-Dihydroxy-androsr-! ,4-diene-3, 17-dione from Acremonium strictum(-preparation of anticanceragent).

Tukovy Prumysl Koncem (1. Novak.B. KraJova. J. Tolman. D. Rysava,J. Smidrkal, V. Lopata, et 01.),Europe Patent 355,228 (1990),Enzymic Agent Based on Lipasesand Oxidoreductases for Washing.Degreasing and Water Recondi-tioning.

Lion (K. Fukano and S. Abe), JapanPatent 2,077.498 and 2,077,499(1990), Lipase-containing Deter-gent Compositions for Dishwash-ing Machines.

S.T. Chemical (M. Minoguchi and T.Muneyuki). Japan Patent 1,285,188(1989). Immobilization of Lipaseon Po1yacryJic Acid and ILSUse inDetergents.

Igene Biotechnology (E.A. Johnson,D. Schreiber, K.P. Ho, W.T. Hall.H.H. Yang and B. Geldiay-Tuncer).U.S.A. Patent 229,536 (1990).Phaffia Yeast with EnhancedAsraxaorhin Content.

Showa Denkc (H. Sato and K. Fukui).Japan Patent 2,086,790 (1990).Manufacture of Tocopherol byPlant Tissue Culture.

Yamanouchi Pharmaceutical (H. Imai,H. Yazawa, K. Nagai, T. Saito andS.F. Liang), WO (Patent Coopera-lion Treaty) 89,009.769 (1989).Substance Q-2819 and Its Prepara-tion from Chrysosporium (--use insuppression of lipid peroxide for-mation).

Pemod-Ricard (R. Cardillo, C. Fugan-ti, G. Sacerdote. M. Barbeni, P.Cabella and F. Squarcia), EuropePatent 356.291 (1990). Microbio-logical Production of (R)-y-Decalactone and (R)-y-Octalactone(-production by Aspergillus niger.Pichia etcneltsu. erc.).

Mitsui Petrochemical (K. Kamisakoand K. Isoi}. Japan Patent2.107.194 (1990), Manufacture of

INFORM, Vol. 2, no. 3 (March 1991)

224

BIOTECHNOLOGY

Scowcroft, BTCI Group of Calgary,Alberta, and R.F. Wilson of North Car-olina State University in Raleigh,North Carolina, will be chairpersons.Scheduled papers include speakersfrom Australia, Canada and the UnitedStales. A complementary session onnew industrial and nutrition uses fornux and linseed oils will feature speak-ers from Canada and the United States.

Two sessions have been organizedby David J. Kyle of Martek Corpora-tion in Columbia, Maryland, on indus-trial applications of single-cell oils.Participating will be speakers fromEngland, Israel, Japan, New Zealand,The Netherlands. Canada and theUnited States.

P.E. Sonnet of the USDA EasternRegional Research Center in Philadel-phia, Pennsylvania. has organized asession on enzyme chemistry for thefats and oils industries. with speakersfrom Canada, Denmark, South Koreaand the United States. A session onbioengineering of fats and oils willhave speakers from England, Japan,The Netherlands and the United States.George Abraham of the USDA South-ern Regional Research Center in NewOrleans. Louisiana, is chairperson.

Finally, there will be a basic tutori-

Triterpencids by Plant Tissue Cul-ture.

Henkel (J. Kretschmann. F.J. Carduck,W.O. Deckwer and C. Tag), Ger-man Patent 3,829,618 (1990), Fer-mentative Manufacture of 1,3-Propanediol from Glycerol.

Nippon Synthetic Chemical (M.Hasegawa and H. Honda), JapanPatent 2,086,786 (1990), MicrobialManufacture of 3(R)-Hydroxybu-tynues.

BASF (H. Rettenmaier, A. Kreimeyer,J. Perner and P. Diessel). GennanPatent 3,834,550 (1990), A Ther-mostable Alkaline Protease fromBacillus for Use in Laundry Deter-gents.

Biotechnology to bemajor topic in ChicagoThe practices and processes ofbiotechnology will receive consider-able attention during the 1991 AOCSAnnual Meeting and Exposition to beheld in Chicago from May 12-15.

A major symposium has beenarranged on biotechnological im-provemeru of flax and linseed oil. WR.

al in plant biotechnology by A.K.Weissinger of North Carolina StateUniversity in Raleigh. R.E Wilson.who arranged for the program,describes Weissinger as "an accom-plished scientist and a dynamicspeaker."

Plant biotechnology encompassesmany scientific disciplines thai have orwill contribute to breakthroughs in theoilseeds industries, Wilson said, andthe area of molecular genetics plays afundamental role in these efforts.

"However, the concepts, tenninolo-gy and experimental approach used inthis science are often foreign to thosetrained in other disciplines," he said."Dr. Weissinger will address, in twoconsecutive 1.5 hour presentations, anintroduction to the vocabulary andprinciples of molecular genetics,applications of RFLP technology, con-cepts in tissue CUlture. in vitro setec-lion processes, genetic rransformanon,and safety issues involved in the over-all development of plums with geneti-cally engineered gene systems."

In addition, a special demonstra-tion of the "gene-gun" ballistic tech-nology developed by DuPonl fOT genetransfer will be exhibited in conjunc-tion with the discussions.

CLEAN COOLING WATER ~

-( - 'u,:~a, ~-Q-

\ i~l~~GI/DEODORIZER I- RECOY~roWER

1t.J..- 1- >-

-~?y ~ \!II ~ ~.~. PU"P DIIAIH

( \!Jro __~.~

TO OIBTILL.ATE IITORAQE

Clean water systems eliminate plant odorsThis deodorizer vacuum system, devel- Get .more information and a quote on

oped b Nash-Kinema for edible-oil proc- upgrading your d~odonzer system. ToeSSing: costs about 28% less 10 o~ate ~_ts3:~~~~~e~A~t~fL~~4~O.d ~~than the greasy-water vacuum systemure- , i76places. Barometric condensers. drop legs, w~ite Nash-Kinema, Inc., P.O. Box ,and hotwellsareeliminated. You solve the Elizabeth, PA., 15037, U.S.A.greasy-waterwaste-mll:"agem~ntproble~. NAS H'Best of all odor-causing residue doesn tcontaminat~yourplant.anditneverreaches I-Ilnemayour cooling tower. EtfGlHEEf'£O \IIICUUM SYSTE/oIS