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Accumulation and Degradation of Polyphosphate in Ac1netobacter Sp. 0000 0284 4245

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Accumulation and Degradation of Polyphosphate in Ac1netobacter Sp. 0000 0284 4245 ACCUMULATION AND DEGRADATION OF POLYPHOSPHATE IN AC1NETOBACTER SP. CENTRALE LANDBOUWCATALOGUS 0000 0284 4245 Promotor: dr A.J.B. Zehnder, hoogleraar in de microbiologic Co-promotor: dr M.H. Deinema, universitair docent ^uo$to\, il'ZT- J.W. van Groenestijn ACCUMULATION AND DEGRADATION OF POLYPHOSPHATE IN ACINETOBACTER SP. Proefschrift ter verkrijging van de graad van doctor in de landbouwwetenschappen, op gezag van de rector magnificus, dr C.C. Oosterlee, in net openbaar te verdedigen op vrijdag 17 juni 1988 des namiddags te vier uur in de aula van de Landbouwuniversiteit te Wageningen BIBLIOTHEEK. LANDBOUWUNIVERSITEIT .WAGENINGEN JO/OO^O'I^^ STELLINGEN 1. Debiochemisch emodelle nva nhe tmechanism eva nd ebio - logischedefosfaterin g vanafvalwater ,zoal sgepresenteer d ophe tinternational econgre sove rdi tproces ,gehoude nva n 28t/ m 30Septembe r 1987t eRome ,kunne nnie tverklare n waaromhe tPhostri pproce sz ogoe dwerkt . -R .Ramador i(ed. )Biologica lphosphat eremova lfro mwast e waters. Proceedingso fa nIAWPR Cspecialize dconference , Rome,28-3 0Sept .1987 .Adv .Wat. Poll .Control .Pergamo n Press. 2. Integenstellin gto td ebewerin gva nKulae vhoef td epro - ductieva nAT Pui textracellulai radenosin ee nintracel ­ lular:polyfosfaa tdoo rafstervend ecelle nva nCorynebac - teriumsp .nie tt eduide no pee npolyfosfaat:AM Pfosfo - transfersaseactiviteit . -V.D . Butukhanov, M.A. Bobyk, V.Zh. Tsyrenov and I.S. Kulaev (1979)Possibl e role of high-molecular-weight polyphosphate inAT Psynthesi sfro mexogenou sadenin eb y acultur eo fCorynebacteriu mspecies ,strai nVSTI-301 . Biokhimiya44:1321-1327 . - I.S.Kulae van dV.M .Vagabo v(1983 )Polyphosphat emeta ­ bolism inmicroorganisms .Adv .Microbiol .Physiol .24:83 - 171. 3. Deconclusi eva nNeijsse le nTempes tda tglucos egelimi - teerdegroe iva nKlebsiell a aerogenesNCT C 418koolsto f gelimiteerdmoe tzij nspreek td ehypothes eva nLinto ne n Stephensontegen . -J.D . Linton and R.J. Stephenson (1978)A preliminary studyo ngrowt h yields in relation to the carbon and energyconten to fvariou sorgani cgrowt hsubstrates .FEM S microbiol.Lett .3:95-98 . -O.M .Neijsse lan dD.W .Tempes t (1976)Th erol eo fenergy - spillingreaction si nth egrowt ho fKlebsiell aaerogene s NCTC41 8i naerobi cchemosta tculture .Arch .Microbiol . 110:305-311. 4. Hethomogenisere nva naktie fsli bmet ee nvortex ,da ti n sommigepublicatie sword tbeschreve nal senig evoorbehan - deling ten behoeve van celtellingen op platen,ka n tot foutieveconclusie s leiden omtrentd e aantallen cellen in ditslib . 5. Zolanggrot ehoeveelhede nfosfaa taanwezi gblijve ni nd e effluentenva nafvalwate rzuiveringsinstallaties , isd e verwijdering vand e laatsteppm' sNH 4 enorganisch ekool - stofverbindingenui tafvalwate rvaa kee nzinloz ebezigheid . Deintroducti eva nfosfaa ti nd eheffingsformul evoo rd e lozingva neffluen ti she tgewenst epolitiek einstrumen t datdez evaa ktoegepast ehandelswijz eka nvoorkomen . 6. Doorhe t introduceren van fosfaatvrije wasmiddelen zal biologischedefosfaterin gva nafvalwate rsteed sgunstige r wordent.o.v .chemisch edefosfatering . 7. Dehoudin gva nregeringe ninzak ed eemissi eva nafgasse ni s eentypisch e'na-mij-de-zonvloed-politiek' . 8. Detijdelijk evolksverontwaardigin g tijdensd eintroducti e vand eeerst etreine ni nEuropa ,word tvaa kte nonrecht e aangehaaldal se rwee ree ncontroversiel etechnologisch e innovatiegeintroduceer dmoe tworden . 9. Natuurtalenten zijnvaa k slechtedocente nomdat zi jnie t inzienwaaro mee nleerlin gd esto fnie tbegrijpt . 10. Vooree nwetenschappe rme tee nachternaa mva nvie rletter - grepenwaari n 'oe'e n 'ij' voorkomen zal internationale bekendheidnie tsne lzij nweggelegd . Stellingen behorende bij het proefschrift "Accumulation and degradationo fpolyphosphat ei nAcinetobacte rsp. "va nJ.W .va n Groenestijn. Wageningen,1 7jun i198 8 VOORWOORD Voordatu i ndi tproefschrif tgaa tleze nmoe tu beseffe nda t de verschillende activiteiten die hebben geleid tot de tot- standkomingva ndi tboe knie the twer ki sgewees tva nee nenke - ling,d eauteur ,maa ri ngrot emat eondersteun dzij ngewees tdoo r anderen.Dez emense nwi li khie rbedanken : Prof.Zehnde re nMi aDeinem avoo rhe tmogelij kmake nva ndi t onderzoek,voo rnu nwetenschappelijk ebegeleiding ,he tkritisc h doorlezenva nhe tmanuscrip te nhu nstimulerend einvloed . De studentenJacque sva nd eWorp ,Gerar dVlekke ,Martin aZuide - ma, Peter Okkerman, Michael Bentvelsen, DesireaAnink , Eric Boschker,Pir nVis ,The oDemmers ,Harri ePeters ,Roelo fRongen , Antoinetteva nMilaan ,Dorin eSijpken se nHedwi gSleiderink ,di e eendirect eo f indirectebijdrag ehebbe ngelever daa nd e inhoud vandi tproefschrift , enwaarme ei kmet vee lplezie rsame nhe b gewerkt. AnsBroersma-D eHaa nvoo rhe ttypwerk . NeesSlotboo mvoo rd etekeningen . FritsLa pvoo rd eelectro-technisch eondersteuning . WimRoelofse nvoo rd eanalytisch eassistentie . Chris Schouten voor de assistentie in de bacteriologische keuken. Hetpersonee lva nd eTFD Lvoo rd eEM-opnamen . Dr.A.J.W.G .Visse r enJ.S .Santem ava nd evakgroe p Biochemie voorhe tmogelij kmake nva nd ebioluminescenti emetingen . Jo Jacobs en Johannes van der Laan van de vakgroep Water- zuiveringvoo rhu nassistenti eme td eAAS-metingen . Envel eander emedewerker sva nd evakgroepe nMicrobiologi ee n Waterzuiveringdi eme traa de ndaa dhebbe nbijgedrage naa nd e totstandkoming van dit proefschrift. Hun gezelschap en be- langstellingvormde nee nhartverwarmend estimulan se nee nbe - langrijkesteunpilaa ronde rhe tsoirt sz ozwar epromotie-werk . Ook mijnouder s komtall edan ktoe :zi jhebbe nmi jgemotiveer d engestimuleer do mt egaa nstuderen . CONTENTS page Chapter1 : Introduction 1 Chapter2 : Regulationo fpolyphosphat eaccumulatio ni n Acinetobactersp . 27 Chapter3 : Therol eo fcation si nth eaccumulatio nan d releaseo fphosphat ei nAcinetobacte r strain210 A 45 Chapter4 : ATPproductio nfro mpolyphosphat ei nAcineto ­ bacterstrai n210 A 63 Chapter5 : Polyphosphatea sa nenerg yreserv ei nviv o inAcinetobacte rstrai n21O A 77 Chapter6 : Polyphosphatedegradin genzyme si nAcineto ­ bactersp .an dactivate dsludg e 93 Chapter7 : Summary+ Samenvattin g 107 CHAPTER 1 INTRODUCTION PREAMBLE A study on polyphosphate accumulation and degradation by Acinetobacter sp.i s important forth e following tworeasons : (1)Polyphosphat e accumulating strains of Acinetobacter play an important role in the biological phosphate removal from waste water, a process that is performed in a number of wastewater treatment plants and results in activated sludge enriched with polyphosphate accumulating bacteria. (2)Acinetobacte r sp. are able to accumulate exceptionally high amounts of phosphorus (up to 100 mg per g dry biomass), mainlyi nth efor m of polyphosphate, and this property makes this bacterium very suitable for research on the reserve functions of polyphosphate. The role of inorganic polyphosphate in biology has not yet been elucidatedver ywell ,especiall y itsrol ea sa n energy reserve invivo ,whic h stillremain s tob ea controversia l subject.I n this chapter the available information about both the appli­ cation of polyphosphate accumulating bacteria in wastewater treatment and the physiological role of polyphosphate in micro­ organisms are summarized. THE ROLE OF PHOSPHATE IN EUTROPHICATION OF SURFACE WATERS Inth e early ninetheenseventies the awareness for water pol­ lution grew exponentially and wastewater treatment plants were introduced on a large scale in the industrialized countries. Thesetreatmen t plants were designed toreduc eth e biological oxygen consuming potential of the waste waters, i.e. organic carbon and reduced nitrogen. The effluents of these plants still may contain largeamount s ofphosphat e and nitrate,whic h are both the major compounds responsible for the eutrophication of surfacewaters .Eutrophication ,a noverloa d of surface waters with nutrients, has become a growing problem, and many negative effects have been reported. Examples are:alga l blooms, decrease of ecological quality and organism diversity in surface waters, and livestock deaths due to ingestion of toxic algae. The treat­ ment of eutrophied water inconventiona lpotabl ewate r treatment plants also presents problems. They include penetration of fil- tersb y algae,filte rclogging ,taste ,odour ,colour ,turbidit y andth eformatio no ftrihalomethane s from organic compounds releasedb yalga ea sa resul to fchlorination .Deat ho falga e canlea dt oanaerobi ccondition si nth ewate rcausin gfis hkil l andodou rnuisance .Excretio no fprotein sb yalga ei nth ese a cancaus e foam whichma y interferwit h fishery.Th e flow of irrigationwate ri ncanal sca nb einterferre db yth eabundan t developmento ffilamentou salga e(Wiecher san dBes t 1985). Ina majorityo fsurfac ewaters ,phosphoru si sth elimitin gnutrient . Itsadditio n tothi swater s will cause eutrophication (Vollen- weider 1985).Accordin gt oVa nDiere ne tal .(1985 )th eeutro ­ phication policy in The Netherlands should consist of four tracks:(1 )phosphat eba nfro mdetergents ,(2 )wate rtreatment (phosphateremova lfro mwast ewater ,dredgin go fsediment scon ­ taining phosphate), (3)restrictio n ofphosphat eemissio nb y agriculture (e.g.runoff )an d (4)reductio no f foreigninflu xo f phosphate (byrivers) . Only an integral approachwil l reduce algalgrowth .Muc hattentio ni sgive nt oth eremova lo fphos ­ phatefro m municipalwast ewater ,whic h iscurrentl y inth e Netherlandsth emos timportan t sourceo fphosphat ea sa pol ­ lutant (Olsthoorn1986) . CHEMICALPHOSPHAT EREMOVA LFRO MWASTEWATE R Theconventiona
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