簡介：PHYSICSPROCEDIA2520122190–219618753892?2012PUBLISHEDBYELSEVIERBVSELECTIONAND/ORPEERREVIEWUNDERRESPONSIBILITYOFGARRYLEEDOI101016/JPHPRO2012033692012INTERNATIONALCONFERENCEONSOLIDSTATEDEVICESANDMATERIALSSCIENCEDATAACQUISITIONANDTRANSMISSIONSYSTEMBASEDONSHORTMESSAGESERVICESUGWONKIM,SLCOCKCROFTABSTRACTTHEDATACOLLECTIONANDTRANSMISSIONSYSTEMBASEDONSINGLECHIPMICROCONTROLLERSCMUSESTHELCDSCREENTODISPLAYTHEREALTIMEDATA,ANDEMPLOYSTHEBUTTONMATRIXASTHEINTERACTIVEINTERFACETOENTERTHEPRESETALARMVALUE,THETARGETCELLPHONENUMBERSANDTHECONTROLSIGNALSTC35IISUSEDASTHEGSMMODULETOTRANSMITDATAANDCONTROLINFORMATIONTHEALARMSHORTMESSAGESMSWILLBESENTAUTOMATICALLYWHENTHEDATAEXCEEDSTHEPRESETALARMVALUEANDUSERALSOCANORDERTHESCMTOSENDTHEREALTIMEDATAANDCOMPLETETHEREMOTECONTROLBYSENDINGASMSOFSPECIFICFORMAT?2011PUBLISHEDBYELSEVIERLTDSELECTIONAND/ORPEERREVIEWUNDERRESPONSIBILITYOFNAMEORGANIZERKEYWORDSSHORTMESSAGESERVICEWIRELESSDATACOLLECTIONTRANSMISSIONSYSTEMWIRELESSMONITORING1INTRODUCTIONATPRESENT,RCDRESIDUALCURRENTDEVICEWIDELYUSEDDOMESTICISTHEELECTRONICONESOFFAULTCURRENTOPERATEDTYPETHEIROPERATIONDEPENDSONTHEDETECTIONANDJUDGMENTOFTHEELECTRICALPARAMETERSOFTHEPROTECTEDCIRCUITBECAUSEOFTHECOMPLICATEDAPPLICATIONCONDITIONOFRCD,THEREAREALOTOFPOTENTIALSAFETYHAZARDSFORTHEPOWERLINESANDABNORMALOPERATIONSARELIKELYTOCAUSEEQUIPMENTDAMAGEANDPROPERTYLOSSESTHEREFORE,AUTOMATICCONTROLANDREMOTEMONITORINGAREREQUIREDTHEOPERATORCANREADILYACQUIRETHEOPERATINGSTATUSOFEQUIPMENTSANDCONTROLTHEMWITHTHECONTINUOUSDEVELOPMENTOFWIRELESSCOMMUNICATIONTECHNOLOGY,THEREISINCREASINGDEMANDSOFINDUSTRIALCONTROLFORTHEWIRELESSMONITORINGWIRELESSTRANSMISSIONTECHNOLOGIESWIDELYUSEDTODAYINCLUDEMICROWAVE,BLUETOOTH,INFRARED,ETCTHETRANSMISSIONDISTANCEOFMICROWAVEISDEPENDENTONTHEWEATHERANDOTHERCONDITIONS,ANDTHECOSTOFESTABLISHINGTHEIROWNBASESTATIONISQUITEHIGHTHUSITSAPPLICATIONONTHEWIRELESSMONITORINGISSUGWONKIM,,WOMAN19646SCHOOLOFINFORMATIONENGINEERING,PROFESSORMASTERINSTRUCTOR,RESEARCHDIRECTIONINTELLIGENTAPPLIANCES,INTELLIGENTCONTROLOFMCUAVAILABLEONLINEATWWWSCIENCEDIRECTCOM?2012PUBLISHEDBYELSEVIERBVSELECTIONAND/ORPEERREVIEWUNDERRESPONSIBILITYOFGARRYLEEOPENACCESSUNDERCCBYNCNDLICENSEOPENACCESSUNDERCCBYNCNDLICENSEADIVISIONOFADVANCEDMATERIALSENGINEERING,RCIT,CHONBUKNATIONALUNIVERSITY,JEONJU561756,REPUBLICOFKOREABDEPARTMENTOFMATERIALSENGINEERING,UNIVERSITYOFBRITISHCOLUMBIA,VANCOUVER,CANADA2192SKIMETALPHYSICSPROCEDIA2520122190–2196FIG2DATAACQUISITIONCIRCUITBGSMMODULETHESIEMENSTC35IISUSEDTOSENDANDRECEIVESHORTMESSAGESASAGSMMODULE,TC35ISERIESARETHEFIRSTCHOICEOFINDUSTRIALCONTROLLEVELFORTHEIRCOSTEFFECTIVEANDUPGRADINGTOGPRSMODULEEASILYTHISMODULECANBECONNECTEDDIRECTLYWITHTHESCMANDALSOSUPPORTSRS232SERIALCOMMUNICATIONTHETESTFORMATANDTODAY’SMAINSTREAMPDUPROTOCOLDATAUNITFORMATCANBEUSEDTOSENDANDRECEIVESMSTHROUGHTHISMODULEINTHISDESIGN,INTERFACESCONNECTEDWITHSCMMAINLYINCLUDEIGTFIREWIRE,RXDANDTXDSIMCARDPINSAREDIRECTLYCONNECTEDWITHTHETC35IPINSOFTHESAMENAMEVIASIMCARDREADERCCINPINOFTHEZIFBLOCKCONNECTORISUSEDTODETECTWHETHERTHESIMCARDISINSERTEDPROPERLYTHELEVELOFCCINPINISHIGHIFTHECONNECTIONISCORRECT,OTHERWISELOWINADDITION,THEPOWERSUPPLYOFTHEMODULESHOULDALSOBEPAIDATTENTIONTOITWILLPOWEROFFAUTOMATICALLYIFTHEVOLTAGEISBELOW33VSINCETHEPOWERWILLBECOMELARGERWHENSENDINGSMS,THEPOWERSUPPLYFORTC35IISGENERALLYSEPARATEDANDTHECURRENTOFTHEPOWERSUPPLYMUSTBEHIGHERTHAN2ATHESCHEMATICDIAGRAMOFTC35IMODULEISSHOWNINFIGURE3FIG3TC35ICIRCUIT

簡介：SEEDISCUSSIONS,STATS,ANDAUTHORPROFILESFORTHISPUBLICATIONATHTTPS//WWWRESEARCHGATENET/PUBLICATION/3780423ANACCONVERTERWITHASMALLDCLINKCAPACITORFORA15KWPERMANENTMAGNETSYNCHRONOUSINTEGRALMOTORCONFERENCEPAPEROCTOBER1998DOI101049/CP19980597SOURCEIEEEXPLORECITATIONS14READS7312AUTHORS,INCLUDINGHPNEEKTHROYALINSTITUTEOFTECHNOLOGY177PUBLICATIONS2,656CITATIONSSEEPROFILEAVAILABLEFROMHPNEERETRIEVEDON27JANUARY2016ROYALINSTITUTEOFTECHNOLOGYCONV10UFSYNTESTCREATEDLASTMODIFIEDPRINTEDONJUNE26,1996ES9545MARCH17,1998KARSTENMARCH17,1998KARSTENSS1GGDD10DDDDDD111100E6100E6100E6000100010001ABCGGDDGDGD000240002400024025025025ABCPHASEFREQMAGSIN7224009ABCOMPARATORABCOMPARATOR100E9100E9ABCOMPARATORABCOMPARATORPHASEFREQMAGSIN1207209ABCOMPARATORABCOMPARATORPHASEFREQMAGSIN72090FIGURE1SCHEMATICDIAGRAMOFTHECONSIDEREDSYSTEM400450500550600650700V01601650170175018SFIGURE2TYPICALSIMULATEDINSTANTANEOUSVALUEOFTHEDCLINKVOLTAGEFSW6KHZ,FOUT72HZASUL?LM√3√2UDC20612MUDC2ATYPICALSIMULATEDINSTANTANEOUSVALUEOFTHEDCLINKVOLTAGEISSHOWNINFIGURE2THERIPPLECONSISTSOFTWOCOMPONENTS–ONEPRODUCEDBYTHERECTIFIERANDONEPRODUCEDBYTHEINVERTERWHICHWILLBEDISCUSSEDINTHENEXTSECTIONIFTHESIZEOFTHECAPACITORISCHANGED,THERIPPLEINTHEDCLINKVOLTAGEWILLBECHANGEDTHEMAXIMUMPEAKTOPEAKVOLTAGEHASASUBSTANTIALDECREASEBETWEENTHECAPACITORSIZES1ΜF(xiàn)AND10ΜF(xiàn)ANDREMAINSALMOSTCONSTANTFORLARGERCAPACITORVALUES,WHICHCANBESEENINFIGURE5USINGACAPACITOROF10ΜF(xiàn)LEADSTOAPEAKTOPEAKVOLTAGERIPPLEOF?UDC290VTHEINVERTEROUTPUTCURRENTISVERYCLOSETOTHESINUSOIDALWAVEFORM,EVENTHOUGHNOCLOSEDLOOPCONTROLWASUSEDATYPICALWAVEFORMOFTHISCURRENTINONEPHASEISSHOWNISFIGURE3ITCONSISTSMAINLYOFTHEDESIREDFUNDAMENTALFREQUENCYFOUT72HZ,SUPERIMPOSEDWITHSMALLRIPPLESCAUSEDBYTHEHIGHFREQUENCYINVERTERPULSES502502550A01601670174018101880195SFIGURE3INVERTEROUTPUTCURRENTANALYTICALEVALUATIONOFTHEDCLINKVOLTAGETHEBASICIDEAOFTHISANALYSISORIGINATEDFROMOBSERVATIONOFTHEINSTANTANEOUSDCLINKVOLTAGEFIGURE2OBTAINEDFROMTHESIMULATIONS4THEVOLTAGEISACOMPOSITIONOFTHERECTIFIEROUTPUTVOLTAGEHAVINGADOMINANTFREQUENCYOF650300HZ,SUPERIMPOSEDWITHTHERIPPLECAUSEDBYTHEPULSEWIDTHMODULATIONOFTHEINVERTERTHEPEAKTOPEAKRIPPLEINTHEDCLINKVOLTAGECAUSEDBYTHERECTIFIERCANBECALCULATEDFROM,?UR√2U??32U3ASSUMINGTHATUISTHELINETOLINERECTIFIERINPUTVOLTAGE5TOGETANANALYTICALEXPRESSIONOFTHEDCLINKVOLTAGERIPPLECAUSEDBYTHEINVERTER,THEINSTANTANEOUSPOWERFLOWINGINTOTHEDCLINKWASCOMPAREDWITHTHEINSTANTANEOUSPOWERFLOWINGOUTOFITDURINGONESWITCHINGINTERVALTHEPHASEOUTPUTVOLTAGESOFTHEINVERTERCONSISTONLYOFTWODISTINCTLEVELS,EITHERUDC2OR?UDC2,WITHRESPECTTOTHEMIDPOINTOFTHEDCLINKVOLTAGEUDISTHEMEANDCLINKVOLTAGE,WHICHISASSUMEDTOBECONSTANTINTHISANALYSISTHEDURATIONOFTHEPULSE,WHERETHEPHASEVOLTAGEHASTHENEGATIVEVALUE?UDC2,NAMEDTP,DEPENDSONTHEINSTANTANEOUSVALUEOFTHESINUSOIDALREFERENCEWAVEFOREACHPHASETHEANALYTICALRELATIONBETWEENTPANDTHEDESIREDINSTANTANEOUSVALUEOFACERTAINOUTPUTPHASEVOLTAGE,UI,CANBEWRITTENASTPITSW2?1?2UIUDC?4FROMTHISPOINTISISPOSSIBLETOCALCULATETHEENERGYDRAWNFROMTHECAPACITORFOREACHSECTIONOFONESWITCHINGINTERVALTSWCALCULATINGTHEENERGIESWIOVERTHEWHOLESWITCHINGINTERVALANDDIVIDINGBYTSW,YIELDSANAVERAGEPOWER,WHICHGIVESRISETOANEQUIVALENTCONTINUOUSCURRENTDRAWNFROMTHECAPACITORTHISPARTISTHEDCPARTOFTHEINSTANTANEOUS

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簡介：2011INTERNATIONALCONFERENCEONELECTRONICSANDOPTOELECTRONICSICEOE2011BASEDONSINGLECHIPMICROCOMPUTERTEMPERATUREANDHUMIDITYDATAACQUISITIONSYSTEMDESIGNXUYANGUOTAOZHULIEKEYLABORATORYOFINSTRUMENTATIONSCIENCEC1,C2ANDC2VALUE,ASSHOWNINTABLEIANDOUTPUTHUMIDITYDATAABOUTMEDIANTABLEIHUMIDITYTRANSFORMCOEFFICIENTSORHC1C2C312BIT400405281068BIT4064872I04°WHENMEASURINGTEMPERATUREWITH25CLARGEDIFFERENCE,INORDERTOCOMPENSATEFORTEMPERATUREOFHUMIDITYMEASUREMENTIMPACT,USETYPE2CORRECTIONOUTPUTVALUERHTRUET25TLT2SORHRH12AMONGTHEMFORRHTRUERH1COMPENSATIONTEMPERATUREINFLUENCEAFTERTHEHUMIDITYVALUE,TLANDT2VALUESASSHOWNINCHART2,ANDOUTPUTHUMIDITYDATADIGITSARECONCERNEDTABLEIITEMPERATURECOMPENSANONCOEFFICIENTSSORHTT212BIT0010000088BIT001000128SHT75BYGAPMATERIALSPTATISPROPORTIONALTOTHEABSOLUTETEMPERATURERDOFTEMPERATURESENSORHASEXCELLENTLINEARITYTHEFOLLOWINGFORMULAWILLBEAVAILABLESHT75DIGITALOUTPUTCONVERSIONFORTEMPERATURETEMPERATUREDLD2SOTAMONGTHEMFORSHT75SOTOUTPUT12OR14TEMPERATUREMEASURINGVALUESD1ANDSHT75WORKINGVOLTAGERELATED,D2ANDOUTPUTOFTEMPERATUREDATADIGITSABOUTSUCHTABLE3SHOWSTABLEIIITEMPERATURECONVERSIONCOEFFICIENTDLVDDD°CDOF5V40404V3975395035V396639353V3960392825V39553923D2DIGITSD￡CD￡F14BIT001001812BIT0040072DTEMPERATUREANDHUMIDITYACQUISITIONSOFTWAREPARTSINTHISPAPERTHESOFTWAREUSEDFORSILICONLABORATORIESIDEPROGRAMISMAINLYTOTHESENSORSOPERATION,THECOLLECTIONTEMPERATUREANDHUMIDITYDATAPROCESSINGANDTHECANCOMMUNICATIONSANDHUMIDITYSENSORRECEIVESPCTOSENDDATATRANSMISSIONCOMMAND,BEGANEXTRACTINGTEMPERATUREANDHUMIDITYCOLLECTEDTOSTOREINC8051F060RAMTHETEMPERATUREANDHUMIDITYOFTHEDATA,THENSTARTTHECANTRANSMISSION,UPWARDPLACEMACHINETRANSMITDATA61SILICONLABSCANWORKBITRATECANREACH1MBIT/SEC,ACTUALRATEMAYBEAFFECTEDBYTHECANBUSONSELECTEDDATATRANSFERPHYSICALLIMITATIONSTHECANPROCESSOR32NEWSOBJECTTHATCANBECONFIGUREDTOSENDORRECEIVEDATAINPUTDATA,MESSAGEOBJECTANDITSLOGOMASKSTOREDINRAMCANMESSAGEALLTHESENDINGANDRECEIVINGDATAFILTERINGAGREEMENTBYCANHANDLEALLFINISHED,NEEDNOTCIPCONTROLLER51INTERVENTION,THISWOULDMAKEFORTHECPUCANCOMMUNICATIONBANDWIDTHISTHESMALLESTCIP51THROUGHSPECIALFUNCTIONREGISTERSCONFIGURATIONCANCONTROLLER,READTHERECEIVEDDATAANDWRITINGREADYTOSENDDATATHECANCONTROLLERCLOCKEQUALSCIP51MCUCLOCKSYSCLKIVTESTANDANALYSISOFTHERESULTSTHEPAPERINTRODUCEDEBUGGINGGOODSYSTEMHASBEENCONDUCTED,ANDSEPARATELYCARRIEDONTHESTATICANDDYNAMICTESTS,COLLECTEDEXPERIMENTALDATAFROMTHEMICROCONTROLLERMEMORYREADOUTDATAFIG3SHOWS,THISSYSTEMCANV2312

簡介：JOURNALOFWINDENGINEERINGANDINDUSTRIALAERODYNAMICS9120031613–1625QUASISTATICWINDLOADCOMBINATIONSFORLOWANDMIDDLERISEBUILDINGSYTAMURAA,,HKIKUCHIB,KHIBIBATOKYOPOLYTECHNICUNIVERSITY,1583IIYAMA,ATSUGI,KANAGAWA2430297,JAPANBRESEARCHINSTITUTEOFTECHNOLOGY,SHIMIZUCORPORATION,3417,ETCHUJIMA,KOTOKU,TOKYO1358530,JAPANABSTRACTFLUCTUATINGSURFACEPRESSUREMEASUREMENTSAREMADEFORFOURFLATROOFEDBUILDINGMODELSWITHSQUAREORRECTANGULARPLANSINORDERTOEXAMINEWINDLOADCOMBINATIONSTHEFLUCTUATINGPRESSURESAREINTEGRATEDOVERALLTHEIRSURFACESANDRESULTSAREOBTAINEDOFALONGWINDFORCE,ACROSSWINDFORCE,VERTICALFORCE,ALONGWINDOVERTURNINGMOMENT,ACROSSWINDOVERTURNINGMOMENTANDTORSIONALMOMENTONTHEIRFRAMESTHEMAXIMUMWINDFORCECOMPONENTANDOTHERSIMULTANEOUSLYOBSERVEDWINDFORCECOMPONENTSAREEXAMINED,ANDSOMEINTERESTINGFACTSREGARDINGTHEWINDFORCECOMBINATIONSAREDISCUSSEDONTHEBASISOFABSOLUTEVALUECORRELATIONS,PHASEPLANEEXPRESSIONSANDSOONTHEMAXIMUMNORMALSTRESSESINTHECOLUMNMEMBERSOFSIMPLEFRAMEMODELSARETHENEXAMINEDINORDERTODIRECTLYCHECKTHEWINDLOADCOMBINATIONEFFECTSR2003ELSEVIERLTDALLRIGHTSRESERVEDKEYWORDSLOWRISEBUILDINGQUASISTATICWINDFORCEALONGWINDFORCEACROSSWINDFORCETORSIONALMOMENTCOMBINEDLOADEFFECTSWINDFORCECORRELATIONINTERNALFORCENORMALSTRESS1INTRODUCTIONTHENECESSITYOFCAPTURINGTHEMAXIMUMWINDFORCESWASFIRSTINTRODUCEDBYDAVENPORT1ASAGUSTFACTORBASEDONTHECONCEPTOFTHEMAXIMUMWINDLOADEFFECTS,DAVENPORT2FIRSTDISCUSSEDTHERELIABILITYOFWINDLOADINGONLOWRISEBUILDINGS,ANDSUGGESTEDSOMEIMPORTANTFACTORSFORITSASSESSMENTHISPAPEREMPHASIZEDTHEIMPORTANCEOFMORESOPHISTICATEDWINDLOADESTIMATIONFORLOWRISEARTICLEINPRESSCORRESPONDINGAUTHORFAX81462429547EMAILADDRESSYUKIOARCHTKOUGEIACJPYTAMURA01676105/SEEFRONTMATTERR2003ELSEVIERLTDALLRIGHTSRESERVEDDOI101016/JJWEIA200309020SIMULTANEOUSLYSAMPLEDEVERY000128SFORTHELOWRISEBUILDINGMODELS,154AND110SAMPLESOF10MINLENGTHINFULLSCALECONVERSIONWEREANALYZEDFORA?14AND16FLOW,RESPECTIVELYFORTHEMIDDLERISEBUILDINGMODELS,28SAMPLESAND7SAMPLESWEREANALYZEDTHETUBINGEFFECTSWERENUMERICALLYCOMPENSATEDBYTHEGAINANDPHASESHIFTCHARACTERISTICSOFTHEPRESSUREMEASURINGSYSTEM10INCIDENTALLY,AHUGENUMBEROFSAMPLESWEREANALYZEDFORTHELOWRISEBUILDINGMODELS,BECAUSEITWASALSOINTENDEDTOUSETHEMTOEXAMINETHEENSEMBLEAVERAGEDINSTANTANEOUSPRESSUREPATTERNSCAUSINGEXTREMEWINDLOADEFFECTS113QUASISTATICWINDLOADFORCESTHEFLUCTUATINGPRESSURESWEREINTEGRATEDTOOBTAINQUASISTATICWINDFORCESFDANDFLVERTICALLIFTFTBASEOVERTURNINGMOMENTSMDANDMLANDBASETORSIONALMOMENTMTTHEYAREEXPRESSEDINNONDIMENSIONALFORMSBASEDONTHEMEANVELOCITYPRESSUREQHATROOFHEIGHT,EG,ALONGWINDFORCECOEFFICIENTCD?FDQHBHACROSSWINDFORCECOEFFICIENTCL?FLQHBHANDTORSIONALMOMENTCOEFFICIENTCMT?MTQHBHRFORWINDDIRECTION0?,WHERER?DB2TD2T122WHENASIMPLEFRAMEWITHACOLUMNATEACHCORNERISASSUMED,THEABOVEWINDFORCECOEFFICIENTSCDCLANDCMTAREPROPORTIONALTOVECTORIALLOADEFFECTSSHEARFORCES,BENDINGMOMENTSINACORNERCOLUMNFORTHEALONGWINDDIRECTION,THEACROSSWINDDIRECTIONANDNORMALTOADIAGONAL,RESPECTIVELYTHUS,THESETHREEQUANTITIESCANBEARTICLEINPRESSTABLE1WINDTUNNELMODELSANDFLOWSMODELBMMDMMHMMLOWRISE,SQUAREPLANLS20020050LOWRISE,RECTANGULARPLANLR17012050MIDDLERISE,SQUAREPLANMS200200200MIDDLERISE,RECTANGULARPLANMR200100200FLOWSCALEPOWERLAWINDEX1/250A?1614FIG1WINDTUNNELMODELYTAMURAETAL/JWINDENGINDAERODYN9120031613–16251615

簡介：ABCDABCD

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簡介：JOURNALOFMATERIALSPROCESSINGTECHNOLOGY1822007418–426THEUSEOFTAGUCHIMETHODINTHEDESIGNOFPLASTICINJECTIONMOULDFORREDUCINGWARPAGESHTANG?,YJTAN,SMSAPUAN,SSULAIMAN,NISMAIL,RSAMINDEPARTMENTOFMECHANICALANDMANUFACTURINGENGINEERING,UNIVERSITIPUTRAMALAYSIA,43400SERDANG,SELANGOR,MALAYSIARECEIVED3SEPTEMBER2004RECEIVEDINREVISEDFORM27JULY2006ACCEPTED10AUGUST2006ABSTRACTPLASTICINJECTIONMOULDINGISONEOFTHEMOSTIMPORTANTPOLYMERPROCESSINGOPERATIONSINTHEPLASTICINDUSTRYTODAYHOWEVER,LACKOFSKILLINMOULDMAKINGANDINJECTIONMOULDINGMACHINECONTROLWILLLEADTODEFECTIVEPLASTICPRODUCTWARPAGEISONETYPEOFDEFECTTHATUSUALLYAPPEARSINPRODUCTSWITHTHICKNESSLESSTHAN1MMTHISPROJECTISGOINGTOFABRICATEAMOULDTHATPRODUCEDATHINPLATEWITHDIMENSION120MM50MM1MMTHETHINPLATEWILLBEUSEDFORWARPAGETESTINGINMOULDFABRICATION,THEMOULDBASETHATPURCHASEWILLBEMACHINEDANDASSEMBLEDAFTERTHAT,THEMOULDISFIXEDONTHEINJECTIONMOULDINGMACHINETHEMACHINESETTINGSHOULDBEMADETOPRODUCETHEPRODUCTTHEN,THEPRODUCTWILLBEUSEDFORTESTINGONTHEEFFECTIVEFACTORSINWARPAGEPROBLEMBYAPPLYINGTHEEXPERIMENTALDESIGNOFTAGUCHIMETHODFROMTHERESULTS,ITSHOWSTHATTHEMOSTEFFECTIVEFACTORONTHEWARPAGEISMELTTEMPERATURETHEFILLINGTIMEONLYSLIGHTLYINFLUENCEDONTHEWARPAGETHEOPTIMUMPARAMETERSTHATCANMINIMIZETHEWARPAGEDEFECTAREMELTTEMPERATURE240?C,FILLINGTIME05S,PACKINGPRESSURE90ANDPACKINGTIME06S?2006ELSEVIERBVALLRIGHTSRESERVEDKEYWORDSPLASTICINJECTIONMOULDTAGUCHIMETHODSEXPERIMENTALDESIGNWARPAGE1INTRODUCTIONMOULDMAKINGISANIMPORTANTSUPPORTINGINDUSTRYBECAUSETHEIRRELATEDPRODUCTSREPRESENTMORETHAN70AMONGTHECOMPONENTSINCONSUMERPRODUCTSTHEHIGHDEMANDFORSHORTERDESIGNANDMANUFACTURINGLEADTIMES,GOODDIMENSIONALITYANDOVERALLQUALITY,ANDRAPIDDESIGNCHANGESHASBECOMETHEBOTTLENECKSINMOULDINDUSTRIES1ITISACOMPLICATEDPROCESS,ANDREQUIREDSKILLEDANDEXPERIENCEDMOULDMAKERGENERALLY,INJECTIONMOULDINGISONEOFTHEMOSTIMPORTANTPOLYMERPROCESSINGOPERATIONSINTHEPLASTICINDUSTRYTODAYAPPROXIMATELYONETHIRDOFALLPLASTICSARECONVERTEDINTOPARTSUSINGINJECTIONMOULDING2THISISONEOFTHEPROCESSESTHATAREGREATLYPREFERREDINMANUFACTURINGINDUSTRYBECAUSEITCANPRODUCECOMPLEXSHAPEPLASTICPARTSWITHGOODDIMENSIONALACCURACYANDVERYSHORTCYCLETIMES3TYPICALEXAMPLESARECASINGSANDHOUSINGSOFTHEPRODUCTSSUCHASCOMPUTERMONITORANDMOBILETELEPHONE,WHICHHAVEATHINSHELLFEATURETHESE?CORRESPONDINGAUTHOREMAILADDRESSSAIHONGENGUPMEDUMYSHTANGPRODUCTSTENDTOBECOMELIGHTER,THINNERANDSMALLERHENCE,THEINTERNALCOMPONENTSOFPRODUCTSHAVETOBEPACKEDINTOHOUSING,WHICHHASSMALLERVOLUMEONEWAYTOINCREASETHESPACEOFHOUSINGPARTSISTOREDUCETHEWALLTHICKNESSHOWEVER,THEINJECTIONMOULDINGOPERATIONBECOMESMOREDIFFICULTASTHEWALLTHICKNESSOFPLASTICPARTSBECOMESTHINNER4THISISBECAUSETHESIGNIFICANTWARPAGEDEFECTWILLBEAPPEAREDTOREDUCETHISSIGNIFICANTDEFECT,TESTINGPROCEDUREREGARDINGTOTHEEFFECTIVEFACTORSISREQUIREDATHINPLATEWITHDIMENSION120MM50MMAND1MMTHICKNESSWILLBEPRODUCEDITISUSEFORTESTINGONTHEEFFECTIVEFACTORSTOMINIMIZETHEWARPAGEDEFECTFIRSTLY,FABRICATINGTHEPLASTICINJECTIONMOULDISNEEDEDAFTERTHAT,THEMOULDISGOINGTOBEASSEMBLEDONTHEINJECTIONMOULDINGMACHINEWHENTHETHINPLATESHAVEBEENPRODUCED,THEYWILLBEUSEDFORTESTINGONTHEEFFECTIVEFACTORSINWARPAGEPROBLEMBYAPPLYINGTHEEXPERIMENTALDESIGNOFTAGUCHIMETHOD2PREPARATIONINFABRICATINGTHEMOULD,SOMEPREPARATIONSARENEEDEDTHECAPABILITYOFTHEMACHINETHATAREAVAILABLEINFACULTYARE09240136/–SEEFRONTMATTER?2006ELSEVIERBVALLRIGHTSRESERVEDDOI101016/JJMATPROTEC200608025420SHTANGETAL/JOURNALOFMATERIALSPROCESSINGTECHNOLOGY1822007418–426FIG3CONFIGURATIONOFCOOLINGCHANNELFORCOREPLATE4MOULDFABRICATIONTHEREARESOMEPARTSINTHEMOULDTHATNEEDTOBEMACHINEDITINVOLVEDHIGHPRECISEANDACCURACYPROCESSHENCE,ITISREQUIREDSKILLEDMOULDMAKERFOLLOWINGISTHEMACHININGPROCESSFOREVERYPARTOFTHEMOULD41TOPCLAMPINGPLATEPART1REAMTHROUGHHOLEWITH16MMDIAMETERDRILLINGMACHINEOPERATIONDESCRIPTIONSTOOLSSTEP1MARKINGTHEPOINTHEIGHTGAUGESTEP2CLAMPINGTHEPLATEONTHEMACHINETABLESTEP3CENTERINGATTHEMARKINGPOINTCENTERDRILLSTEP4DRILLINGTHROUGHHOLE?155DRILLSTEP5REAMINGTHEHOLE?16REAMERTOOL,HIGHSPEEDSTEELSPINDLESPEED,330RPMPART2TAPM6HOLESDRILLINGMACHINEOPERATIONDESCRIPTIONSTOOLSSTEP1MARKINGTHEPOINTHEIGHTGAUGESTEP2CLAMPINGTHEPLATEONTHEMACHINETABLESTEP3CENTERINGATTHEMARKINGPOINTCENTERDRILLSTEP4DRILLING2HOLESWITH12MMDEPTH?5DRILLSTEP5TAPPINGTHEHOLESM6TAPPERTOOL,HIGHSPEEDSTEELSPINDLESPEED,1000RPMPART3ENLARGEHOLETO40MMDIAMETERWITH148MMDEPTHMILLINGMACHINEOPERATIONDESCRIPTIONSTOOLSSTEP1MARKING?35AND?40CIRCLESCOMPASSSTEP2CLAMPINGTHEPLATEONTHEMACHINETABLESTEP3MILLINGTHEAREAAT?35CIRCLEWITH148MMDEPTH?10ENDMILLSTEP4BORINGUNTIL?40CIRCLEWITH148MMDEPTHBOREBITTOOL,HIGHSPEEDSTEELSPINDLESPEED,820RPMDEPTHOFCUT,2MMFORROUGHINGAND08MMFORFINISHINGFIG4SHOWSTHEPARTSOFTHETOPCLAMPINGPLATETHATNEEDTOBEMACHINEDNOTETHATTHEFIRSTSTEPINPART1,PART2ANDPART3FIG4PARTSOFTHETOPCLAMPINGPLATETHATNEEDTOMACHINEWHICHAREMARKINGPOINTANDMARKINGCIRCLEISDONEATTHESAMETIMEBEFOREPROCEEDTONEXTSTEP42CAVITYPLATEPART1REAMTHROUGHHOLEWITH16MMDIAMETERDRILLINGMACHINEOPERATIONDESCRIPTIONSTOOLSSTEP1MARKINGTHEPOINTHEIGHTGAUGESTEP2CLAMPINGTHEPLATEONTHEMACHINETABLESTEP3CENTERINGATTHEMARKINGPOINTCENTERDRILLSTEP4DRILLINGTHROUGHHOLE?155DRILLSTEP5REAMINGTHEHOLE?16REAMERTOOL,HIGHS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簡介：AFRICANJOURNALOFBIOTECHNOLOGYVOL821,PP58075813,2NOVEMBER,2009AVAILABLEONLINEATHTTP//WWWACADEMICJOURNALSORG/AJBISSN1684–5315?2009ACADEMICJOURNALSFULLLENGTHRESEARCHPAPERSEPARATIONANDRECOVERYOFORGANICACIDSFROMFERMENTEDKITCHENWASTEBYANINTEGRATEDPROCESSFARAHNADIAOMAR,NOR’AINIABDULRAHMAN,HALIMATUNSAADIAHHAFID,PHANGLAIYEEANDMOHDALIHASSANDEPARTMENTOFBIOPROCESSTECHNOLOGY,FACULTYOFBIOTECHNOLOGYANDBIOMOLECULARSCIENCES,UNIVERSITYPUTRAMALAYSIA,43400SERDANG,SELANGOR,MALAYSIAACCEPTED4SEPTEMBER,2009ORGANICACIDSPRODUCEDFROMANAEROBICDIGESTIONOFKITCHENWASTEWERERECOVEREDUSINGANEWINTEGRATEDMETHODWHICHCONSISTEDOFFREEZINGANDTHAWING,CENTRIFUGATION,FILTRATIONANDEVAPORATIONTHEMAINORGANICACIDPRODUCEDWASLACTICACID98AFTERTHEFREEZINGANDTHAWINGPROCESS,73OFTHETOTALSUSPENDEDSOLIDSWEREREMOVEDANDTHEORGANICACIDSWEREELEVATEDFROM590TO70G/LTHEEVAPORATIONTECHNIQUEWASUSEDTOFURTHERCONCENTRATETHEORGANICACIDSUPTO224G/LUSINGTHEINTEGRATEDRECOVERYMETHOD,THEREDUCTIONOFTHETOTALSUSPENDEDSOLIDSINTHESOLUTIONACHIEVEDWASABOUT93THEMATERIALBALANCEFORTHERECOVERYPROCESSWASALSOPRESENTEDKEYWORDSANAEROBICDIGESTION,KITCHENWASTE,ORGANICACIDS,RECOVERYINTRODUCTIONTHEREMARKABLEGROWTHECONOMYINMALAYSIAHASBROUGHTTREMENDOUSPOPULATIONGROWTHWHICHTHENRESULTEDINAHUGEAMOUNTOFMUNICIPALSOLIDWASTEMSWBEINGGENERATEDTHROUGHOUTTHEYEARMSWISUSUALLYDUMPEDINLANDFILLSANDDEGRADEDINTOSIMPLERCOMPOUNDSHOWEVER,DUETOTHESCARCITYOFLANDANDTHECONSTRAINTOFLANDFILLAREAS,ALTERNATIVEMETHODSSHOULDBECONSIDEREDINORDERTOREDUCETHESPACENEEDEDFORLANDFILLSINCINERATIONISALSOANOPTIONOHKOUCHIANDINOUE,2006ASITISANEASYFINALDISPOSALMETHODBUTLARGEAMOUNTSOFENERGYWILLBEWASTEDINTHEBURNINGPROCESSTSAI,2008SINCE2254OFMSWCONSISTSOFFOODANDORGANICWASTESKATHIRVALEETAL,2003,BIOCONVERSIONOFMSWINTOUSEFULPRODUCTSISFEASIBLEANDITSTREATMENTCOSTCANBEREDUCEDTOOFOODWASTECONTAINSABOUT80OFMOISTUREWHICHISEASILYBIODEGRADABLEANDISRICHINNUTRIENTSANDMICROFLORAWANGETAL,2001DUETOTHESECHARACTERISTICS,BIOLOGICALTREATMENTSAREPREFERREDONEOFTHEMOSTPRACTICALBIOLOGICALTREATMENTSISANAEROBICDIGESTIONITISAGREATTOOLTOSTABILIZEALARGEVOLUMEOFWASTEMATERIALSECONOMICALLYANDEFFECTIVELYSAKAIETAL,2000ITALSOCORRESPONDINGAUTHOREMAILNOR_AINIBIOTECHUPMEDUMYTEL60389466669FAX60389467510PRODUCESLOWBIOMASSANDISABLETODESTROYPATHOGENSBANARJEEETAL,1999FOODWASTECOULDBEANAEROBICALLYFERMENTEDTOPRODUCEVOLATILEFATTYACIDS,ALDEHYDES,ALCOHOLSANDCARBONDIOXIDESTABNIKOVAETAL,2006RATHERTHANBEINGDEGRADEDANDUNEXPLOITEDINTHELANDFILLS,FOODWASTECANBEUTILIZEDFORMANYAPPLICATIONSSUCHASTHEPRODUCTIONOFBIOPLASTICSSAKAIETAL,2004,BIOGASZHUETAL,2008,BIOHYDROGENANDETHANOLPRODUCTIONSKIMETAL,2004,COMPOSTINGTSAI,2008ADHIKARIETAL,2008ANDORGANICACIDSPRODUCTIONOHKOUCHIANDINOUE,2007ZHANGETAL,2008ORGANICACIDSAREORGANICCOMPOUNDSWHICHHAVEBEENEXTENSIVELYUSEDINTHEFOODANDBEVERAGESINDUSTRIES,PHARMACEUTICALINDUSTRIES,COSMETICS,ANDDETERGENTSANDRECENTLYTHEYAREEXTENSIVELYUSEDFORBIOPOLYMERPRODUCTIONSAUERETAL,2008THEREARETWOWAYSTOPRODUCEORGANICACIDSWHICHARECHEMICALSYNTHESISORBIOLOGICALSYNTHESISALTAFETAL,2007WHETHERITISCHEMICALLYORBIOLOGICALLYSYNTHESIZED,THECRITICALSTAGEOFTHEPROCESSISPRODUCTRECOVERYDOWNSTREAMPROCESSESINCLUDINGSEPARATION,EXTRACTIONANDPURIFICATIONOFORGANICACIDSWHICHCONTRIBUTEMORETHAN60OFTHEOVERALLPRODUCTIONCOSTHANETAL,2000SEVERALMETHODSHAVEBEENREPORTEDFOREXTRACTINGORGANICACIDSFROMTHEFERMENTATIONBROTHSUCHASELECTRODIALYSISHUANGETAL,2007,MEMBRANEFILTRATIONKANGANDCHANG,2005,SOLVENTEXTRACTIONHARINGTONOMARETAL5809FIGURE2ORGANICACIDSPROFILEIN50LBIOREACTORTOTALORGANICACIDS?,LACTICACID?,ACETICACID?ANDFORMICACID?RESULTSAREMEANSFORDUPLICATEEXPERIMENTSFIGURE3PROFILEOFPH?ANDBACTERIADISTRIBUTION?DURINGKITCHENWASTEFERMENTATIONRESULTSAREMEANSFORDUPLICATEEXPERIMENTSSHOWNAFTERSEVENDAYSOFFERMENTATION,TOTALSUSPENDEDSOLIDSWEREREDUCEDAT62ORGANICACIDSRECOVERYANDTOTALSUSPENDEDSOLIDSREDUCTIONFIGURE4SHOWSTHEORGANICACIDS,TOTALSUSPENDEDSOLIDSANDTOTALKJELDAHLNITROGENOBTAINEDATEACHUNITOPERATIONOFTHERECOVERYPROCESSTHERECOVERYPROCESSINCLUDESPHYSICALSEPARATIONTHATISFREEZINGANDTHAWING,CENTRIFUGATIONANDFILTRATIONWHILETHEEVAPORATIONSTEPISAMETHODUSEDTOCONCENTRATETHERECOVEREDORGANICACIDSBYELIMINATINGEXCESSWATERTHEORGANICACIDSCONTENTINCREASEDBYABOUT56WHILETHETOTALSUSPENDEDSOLIDSDECREASEDBYABOUT622AFTERTHEFERMENTATIONBYTHEFREEZINGANDTHAWINGPROCESS,THEORGANICACIDSCONTENTINCREASEDBY16WITH66REMOVALOFTOTALSUSPENDEDSOLIDSTHECENTRIFUGATIONANDFILTRATIONPROCESSDONOTHAVEANEFFECTONTHECONCENTRATIONOFTHEORGANICACIDSTHETOTALSUSPENDEDSOLIDSDECREASEDFROM1789TO1227G/LWATERWASEVAPORATEDAT50°CINVACUUMANDTHEREMAININGORGANICACIDSANDSOMESUSPENDEDSOLIDSWERECONCENTRATEDAFTERTHEEVAPORATIONSTEP,BOTHTHEORGANICACIDSANDTHETOTALSUSPENDEDSOLIDSCONTENTSWEREINCREASEDBY69AND83,RESPECTIVELYTHENITROGENCONTENTOFTHEFERMENTATIONBROTHANDRECOVEREDLIQUIDMEDIUMWEREANALYZEDINTHISSTUDYTHE

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簡介：ABSTRACTTHEASYMMETRICREDUCTIONOFETHYL4CHLORO3OXOBUTANOATECOBETOETHYLS4CHLORO3HYDROXYBUTANOATESCHBEWASINVESTIGATEDESCHERICHIACOLICELLSEXPRESSINGBOTHTHECARBONYLREDUCTASES1GENEFROMCANDIDAMAGNOLIAEANDTHEGLUCOSEDEHYDROGENASEGDHGENEFROMBACILLUSMEGATERIUMWEREUSEDASTHECATALYSTINANORGANICSOLVENTWATERTWOPHASESYSTEM,SCHBEFORMEDINTHEORGANICPHASEAMOUNTEDTO258M430G/L,THEMOLARYIELDBEING85ECOLITRANSFORMANTCELLSCOPRODUCINGS1ANDGDHACCUMULATED125M208G/LSCHBEINANAQUEOUSMONOPHASESYSTEMBYCONTINUOUSLYFEEDINGONCOBE,WHICHISUNSTABLEINANAQUEOUSSOLUTIONINTHISCASE,THECALCULATEDTURNOVEROFNADPTHEOXIDIZEDFORMOFNICOTINAMIDEADENINEDINUCLEOTIDEPHOSPHATETOCHBEWAS21,600MOL/MOLTHEOPTICALPURITYOFTHESCHBEFORMEDWAS100ENANTIOMERICEXCESSINBOTHSYSTEMSTHEAQUEOUSSYSTEMUSEDFORTHEREDUCTIONREACTIONINVOLVINGECOLIHB101CELLSCARRYINGAPLASMIDCONTAININGTHES1ANDGDHGENESASACATALYSTISSIMPLEFURTHERMORE,THESYSTEMDOESNOTREQUIRETHEADDITIONOFCOMMERCIALLYAVAILABLEGDHORANORGANICSOLVENTTHEREFORETHISSYSTEMISHIGHLYADVANTAGEOUSFORTHEPRACTICALSYNTHESISOFOPTICALLYPURESCHBEINTRODUCTIONOPTICALLYACTIVE4CHLORO3HYDROXYBUTANOICACIDESTERSAREUSEFULCHIRALBUILDINGBLOCKSFORTHESYNTHESISOFPHARMACEUTICALSTHERENANTIOMERISAPRECURSOROFLCARNITINEZHOUETAL1983,ANDSENANTIOMERISANIMPORTANTSTARTINGMATERIALFORHYDROXYMETHYLGLUTARYLCOAHMGCOAREDUCTASEINHIBITORSKARANEWSKYETAL1990MANYSTUDIESHAVEDESCRIBEDTHEMICROBIALORENZYMATICASYMMETRICREDUCTIONOF4CHLORO3OXOBUTANOICACIDESTERSARAGOZZINIANDVALENTI1992BAREETAL1991HALLINANETAL1995PATELETAL1992SHIMIZUETAL1990WONGETAL1985BASEDONTHEREDUCTIONBYBAKER’SYEASTZHOUETAL1983WEHAVEPREVIOUSLYSHOWEDTHATCANDIDAMAGNOLIAEAKU4643CELLSREDUCEDETHYL4CHLORO3OXOBUTANOATECOBETOSCHBEWITHANOPTICALPURITYOF96ENANTIOMERICEXCESSEEYASOHARAETAL1999ASTHISYEASTHASATLEASTTHREEDIFFERENTSTEREOSELECTIVEREDUCTASESWADAETAL1998,1999A,B,THESCHBEPRODUCEDBYTHISYEASTWASNOTOPTICALLYPUREFROMAMONGTHESETHREEENZYMES,ANNADPHDEPENDENTCARBONYLREDUCTASE,DESIGNATEDASS1,WASPURIFIEDANDCHARACTERIZEDINSOMEDETAILWADAETAL1998WECLONEDANDSEQUENCEDTHEGENEENCODINGS1ANDOVEREXPRESSEDITINESCHERICHIACOLICELLSTHISECOLITRANSFORMANTREDUCEDCOBETOOPTICALLYPURESCHBEINTHEPRESENCEOFGLUCOSE,NADP,ANDCOMMERCIALLYAVAILABLEGLUCOSEDEHYDROGENASEGDHASACOFACTORGENERATORYASOHARAETAL2000HERE,WEDESCRIBETHECONSTRUCTIONOFTHREEECOLITRANSFORMANTSCOEXPRESSINGTHES1FROMCMAGNOLIAEANDGDHFROMBACILLUSMEGATERIUMGENESANDANALYZETHEREDUCTIONOFCOBECATALYZEDBYTHESESTRAINSPREVIOUSREPORTSONTHEENZYMATICREDUCTIONOFCOBETORCHBEWITHANOPTICALPURITYOF92EEKATAOKAETAL1999SHIMIZUETAL1990RECOMMENDEDANORGANICSOLVENTTWOPHASESYSTEMREACTIONFORANENZYMATICORMICROBIALREDUCTION,BECAUSETHESUBSTRATECOBEISUNSTABLEINANAQUEOUSSOLVENTANDINACTIVATESENZYMESWEEXAMINEDTHEREDUCTIONOFCOBETOOPTICALLYPURENKIZAKI?YYASOHARAJHASEGAWAFINECHEMICALSRESEARCHLABORATORIES,KANEKACORPORATION,1–8MIYAMAE,TAKASAGO,HYOGO676–8688,JAPANEMAILNORIYUKIKIZAKIKANEKACOJPTEL81794452415,FAX81794452668MWADAMKATAOKASSHIMIZUDIVISIONOFAPPLIEDLIFESCIENCES,GRADUATESCHOOLOFAGRICULTURE,KYOTOUNIVERSITY,KITASHIRAKAWA,SAKYOKU,KYOTO616–8502,JAPANPRESENTADDRESSMWADA,DEPARTMENTOFBIOSCIENCE,FUKUIPREFECTURALUNIVERSITY,4–11KENJYOJIMA,MATSUOKACHO,FUKUI910–1195,JAPANAPPLMICROBIOLBIOTECHNOL200155590–595DOI101007/S002530100599ORIGINALPAPERNKIZAKIYYASOHARAJHASEGAWAMWADAMKATAOKASSHIMIZUSYNTHESISOFOPTICALLYPUREETHYLS4CHLORO3HYDROXYBUTANOATEBYESCHERICHIACOLITRANSFORMANTCELLSCOEXPRESSINGTHECARBONYLREDUCTASEANDGLUCOSEDEHYDROGENASEGENESRECEIVED7JUNE2000/RECEIVEDREVISION11NOVEMBER2000/ACCEPTED8DECEMBER2000/PUBLISHEDONLINE7APRIL2001?SPRINGERVERLAG2001592VARIOUSCONCENTRATIONSOFCHBE,WASINCUBATEDAT30°CFOR24HINORDERTOSTUDYTHEENZYME’SSTABILITYINTHEPRESENCEOFCHBETHEREMAININGENZYMEACTIVITIESWEREASSAYEDASDESCRIBEDABOVECOBEREDUCTIONWITHECOLICELLSEXPRESSINGTHES1GENEANDECOLICELLSEXPRESSINGGDHGENESINATWOPHASESYSTEMREACTIONTHEREACTIONMIXTURECOMPRISED15MLCULTUREBROTHOFECOLIHB101CARRYINGPNTG,17MLCULTUREBROTHOFECOLIHB101CARRYINGPNTS1,16MGNADP,4GGLUCOSE,25GCOBE,25MLNBUTYLACETATE,ANDABOUT25MGTRITONX100THEPHOFTHEREACTIONMIXTUREWASCONTROLLEDAT65WITH5MSODIUMHYDROXIDEAT2H,125GCOBEAND25GGLUCOSEWEREADDEDTOTHEREACTIONMIXTURETOCOMPARETHEREACTIONBYECOLITRANSFORMANTCOEXPRESSINGTHEGDHANDS1GENES,30MLCULTUREBROTHOFECOLIHB101CARRYINGPNTS1GWASUSEDINSTEADOFCULTUREBROTHOFECOLIHB101CARRYINGPNTGANDECOLIHB101CARRYINGPNTS1OTHERCOMPONENTSANDTHEPROCEDUREWERETHESAMEASDESCRIBEDABOVECOBEREDUCTIONTOSCHBEINATWOPHASESYSTEMREACTIONTHEREACTIONMIXTURECONTAINED50MLOFCULTUREBROTHOFANECOLIHB101TRANSFORMANT,32MGNADP,11GGLUCOSE,10GCOBE,50MLNBUTYLACETATE,ANDABOUT50MGTRITONX100THEREACTIONMIXTUREWASSTIRREDAT30°C,ANDTHEPHWASCONTROLLEDAT65WITH5MSODIUMHYDROXIDEFIVEGRAMSOFCOBE/55GGLUCOSEAND10GCOBE/11GGLUCOSEWEREADDEDTOTHEREACTIONMIXTUREAT3HAND7H,RESPECTIVELY32MGNADPWASADDEDAT26HCOBEREDUCTIONTOSCHBEINANAQUEOUSSYSTEMREACTIONTHEREACTIONMIXTUREWASMADEUPOF50MLOFCULTUREBROTHOFANECOLIHB101TRANSFORMANT,31MGNADP,11GGLUCOSE,ANDABOUT50MGTRITONX100THEREACTIONMIXTUREWASSTIRREDAT30°CFIFTEENGRAMSOFCOBEWASFEDCONTINUOUSLYBYMEANSOFAMICROFEEDINGMACHINEATARATEOFABOUT002G/MINFORABOUT12HTHEPHOFTHEREACTIONMIXTUREWASCONTROLLEDAT65WITH5MSODIUMHYDROXIDETHEREACTIONMIXTUREWASEXTRACTEDWITH100MLETHYLACETATETHEORGANICLAYERWASDRIEDOVERANHYDROUSSODIUMSULFATEANDTHENEVAPORATEDINVACUOANALYSISTHEORGANICLAYERWASOBTAINEDONCENTRIFUGATIONOFTHEREACTIONMIXTUREANDWASASSAYEDFORCHBEANDCOBEBYGASCHROMATOGRAPHYOPTICALPURITYOFCHBEWASANALYZEDBYHIGHPERFORMANCELIQUIDCHROMATOGRAPHYHPLC,ASDESCRIBEDPREVIOUSLYYASOHARAETAL1999ENZYMESANDCHEMICALSRESTRICTIONENZYMESANDDNAPOLYMERASEWEREPURCHASEDFROMTAKARASHUZOJAPANCOBEMOLECULARWEIGHT16459WASPURCHASEDFROMTOKYOKASEIKOGYOJAPANRACEMICCHBEMOLECULARWEIGHT16660WASSYNTHESIZEDBYREDUCTIONOFCOBEWITHNABH4ALLOTHERCHEMICALSUSEDWEREOFANALYTICALGRADEANDCOMMERCIALLYAVAILABLERESULTSCONSTRUCTIONOFECOLITRANSFORMANTSOVERPRODUCINGS1ANDGDHTOEXPRESSTHECARBONYLREDUCTASES1ANDGDHGENESINTHESAMEECOLICELLS,FOUREXPRESSIONVECTORSWERECONSTRUCTEDFIG1PLASMIDSPNTS1GANDPNTGS1CONTAINTHES1GENEFROMCMAGNOLIAE,THEGDHGENEFROMBMEGATERIUM,THELACPROMOTERDERIVEDFROMPUC19,ANDTHETERMINATORDERIVEDFROMPTRC99APLASMIDPNTS1CONTAINSTHES1GENE,THELACPROMOTERDERIVEDFROMPUC19,ANDTHETERMINATORDERIVEDFROMPTRC99ATHEENZYMEACTIVITIESINCELLFREEEXTRACTSOFTHEECOLITRANSFORMANTSARESHOWNINTABLE1ECOLIHB101CELLSCARRYINGTHEVECTORPLASMIDPUCNTHADNODETECTABLES1ORGDHACTIVITYECOLIHB101CARRYINGEITHERPNTS1GORPNTGS1SHOWEDS1ANDGDHACTIVITYWITHOUTISOPROPYLΒDTHIOGALACTOPYRANOSIDEIPTGINDUCTIONTHES1ACTIVITIESOFTHESETWOTRANSFORMANTSWERELOWERTHANTHEGDHACTIVITIESTOOBTAINATRANSFORMANTWHOSES1ACTIVITYWASEQUALTOORGREATERTHANTHELEVELOFGDHACTIVITY,WEUSEDALOWERCOPYVECTOR,PSTV28HOMMAETAL1995TAKAHASHIETAL1995,TOEXPRESSTHEGDHGENEITMAYBEPOSSIBLETORAISETHES1ACTIVITYBYLOWERINGTHEGDHACTIVITYPLASMIDPSTVGCONTAINSTHEGDHGENE,THELACPROMOTER,THECHLORAMPHENICOLRESISTANCEGENE,ANDTHEREPLICATIVEORIGINDERIVEDFROMPACYC184FORCOMPATIBILITYWITHTHEPLASMIDPNTS1INECOLIHB101CARRYINGPNTS1ANDPSTVG,THES1ACTIVITYWASHIGHERTHANTHEGDHACTIVITY,BUTTHISGDHLEVELMAYBETOOLOWTOREGENERATEINACOBEREDUCTIONREACTIONASDESCRIBEDBELOWSTUDYOFENZYMESTABILITYTHESTABILITYOFS1INTHEPRESENCEOFVARIOUSORGANICSOLVENTSISSHOWNINTABLE2S1WASSTABLEINTHEPRESENCEOFNBUTYLACETATE,1OCTANOL,ANDISOPROPYLETHERGDHTABLE1S1ANDGDHACTIVITIESINCELLFREEEXTRACTSOFECOLITRANSFORMANTSPLASMIDTRANSFORMEDSPECIFICACTIVITYU/MGS1GDHPUCNT001001PNTS1160001PNTG001121PNTS1G115772PNTGS1490117PNTS1ANDPSTVG135160TABLE2STABILITYOFS1INVARIOUSORGANICSOLVENTSFOROTHERCONDITIONS,SEETHETEXTORGANICSOLVENTRELATIVEREMAININGACTIVITYNONE771ETHYLACETATE22NBUTYLACETATE3111OCTANOL560CHLOROFORM0DICHLOROMETHANE0ISOPROPYLETHER348TOLUENE0