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1、2005年12月ABSTRACTAtpresent,thebiologicaltreatmenttechnologyisinthetransitionalperiodfrommixedculturetopureculture．Inthispaper,twostrainstodegradephenolwithhighactivitywe犯isolatedandidentifiedfromacclimatedactivatedsludgebyusingthetraditionalmicroorganismbreedingtechnology．Inbetween,theyeastCa
2、ndidatropicaliswascompletelyinvestigatedforitscharactersofphenoliccompoundsbiodegradation．InordertoenhancethecapacitiesofCtropwalistodegradephenoliccompounds，He-NelaserWasutilizedtoirradiatewild-typeCtropicalis．Theoptimalirradiationparameterswei'eobtainedthroughalotofparallelexperimentsasfol
3、lowing：10roWl0—15min,12．5mWl0-15minandl5roWlorain．Correspondingly,highpositivemutationfrequencywasobtainedinthisrange．Inphenolbiodegradationtests，70positivemutantswithhi【ghphenolbiodegradationpotentialWerescreened．PhenolbiodegradationstabilitieswereinvestigatedandCTM2wasattainedandusedinthef
4、ollowingstudiesforitsforemostbiodagradationpotentialandstability．Becauseofthedefinitiveeffectsofphenolhydroxylaseandcatecholdioxygenaseonphenolmetabolism，theactivitiesofthesetwoerlzymeswereinvestigated．ItshowedtheactivitiesofthesetwoenzymeswerehigherinmutantstrainCTM2thaninthewildstrain,whic
5、hwaspresumablyduet0thelaserirradiation．Thus，thefull-lengthsequenceofphenolhydroxylasegeneWassucecssfullyclonedbyPCRmethodinwildandmutantCtropicalis．respectively．Thecomparisonofaminoacidofphenolhydroxylasebetweenwild-typeslrainandmutantstrainCTM岔illustratedfouraminoacidsweremutatedafterthestr
6、ainWasirradiatedbyHe-Nelaser．Inaddition,heterogenousexpressionofphenolhydroxylasevalidatedthetruththattheenzymeactivityvariedaftercellirradiatedbylaser．Thecharactersofphenol，m-cresoland4一chlorophenolbiodegradationwerecontrastivelystudiedbetweenmutantstrainCTM2anditsparentstrain．Theirmaximump
7、henolbiodegradationswere2000and2600rng／L，andthemaximumspecificgrowthrateswereO．48and0．54I／b,respectively．Besides．thebiodegradationofthewildandmutantstrainswerealSOstudiedindual．substratesystem。respectively．Theexperimentsshowedthattheinhibitionstoee
