VPLYV VYBRANÝCH FAKTOROV NA BIOPRÍSTUPNOSŤ ORGANICKÝCH POLUTANTOV A ICH BIODEGRADÁCIU V PÔDNYCH SYSTÉMOCH
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1 ACTA ENVIRONMENTALICA UNIVERSITATIS COMENIANAE (BRATISLAVA) Vol. 15, 2(2007): ISSN VPLYV VYBRANÝCH FAKTOROV NA BIOPRÍSTUPNOSŤ ORGANICKÝCH POLUTANTOV A ICH BIODEGRADÁCIU V PÔDNYCH SYSTÉMOCH Martin Urík 1, Jaroslav Ševc 1, Slavomír Čerňanský 2, Pavol Littera 2, Marek Kolenčík 1 Univerzita Komenského v Bratislave, Prírodovedecká fakulta, 1 Geologický ústav, 2 Katedra ekosozológie a fyziotaktiky, Mlynská dolina, Bratislava, urik@fns.uniba.sk Abstract: Influence of different factors on bioavailability of organic pollutants and their biodegradation in solid substrates. In this work we have reviewed some of the factors affecting bioavailability of organic pollutants in soil systems in connection with their biodegradation, especially sorption of organic pollutants, micropore exclusion of microorganisms and solubility of pollutants. Kľúčové slová: bioprístupnosť, biodegradácia, organické polutanty ÚVOD Bioprístupnosť je jedným z najdôležitejších faktorov, ktorý limituje rozsah a rýchlosť biodegradácie organických polutantov, vrátane chlórovaných uhľovodíkov, aromatických zlúčenín alebo organických látok ropného pôvodu. Napriek tomu, že celkový obsah polutantu v tuhých substrátoch môže byť vysoký, jeho bioprístupný obsah je natoľko nízky, že neaktivuje biodegradačnú aktivitu mikroorganizmov. V prípade, že je polutant pre mikroorganizmy hlavným energetickým zdrojom, predlžuje jeho nedostupnosť lag fázu mikrobiálnych spoločenstiev. Obmedzená bioprístupnosť polutantu je jedným z možných dôvodov zlyhania efektívnych, laboratórne overených bioremediačných technológií. Cieľom príspevku je prezentovať základné zhodnotenie faktorov limitujúcich bioprístupnosť organických polutantov v pôdnych systémoch. Sú to predovšetkým sorpčné reakcie, rozpustnosť polutantu a mikropórová exklúzia mikroorganizmov. VPLYV SORPCIE NA BIODEGRADÁCIU Organické polutanty môžu byť v pôdnych systémoch rozpustené v pôdnom roztoku alebo sorbované v pôdnych časticiach (Shen 1999). Sorpcia 115
2 zahŕňa rôzne medzifázové transportné procesy polutantov (absorpcia, adsorpcia) v pôde alebo vodonosnom horizonte. Pôdne častice sú tvorené anorganickou a organickou frakciou. Anorganickú frakciu tvoria rôzne minerály, ktoré sú viazané v agregátoch polyvalentnými katiónmi alebo organickými látkami (Barra et al. 1998). Pôdne minerály nesú na svojom povrchu pozitívny alebo negatívny náboj. Preto prioritne reagujú s látkami polárneho charakteru alebo s ionizovanými funkčnými skupinami polutantu. Pôdne organické látky sú veľmi pestrou zložkou s rôznou polaritou, kyslosťou, aromatickosťou a stupňom kondenzácie v závislosti od ich zdroja a stupňa degradácie (Pettersson 1994; Mobed et al. 1996). Sorpciu polutantov na anorganickú alebo organickú frakciu ovplyvňuje jeho náboj alebo polarita. Neutrálne molekuly polutantu sa primárne sorbujú na organické frakcie, zatiaľ čo nabité alebo vysoko polárne molekuly sa viažu na povrchy minerálov opačného náboja alebo polarity. Sorpcia slabo polárnych a nepolárnych organických polutantov je preto závislá najmä od podielu organických látok v pôde (Iraqi et Iraqi 2000; Berglöf et al. 2002; Spark et Swift 2002), ale aj od sily väzby polutantu so sorpčnými pozíciami organických látok (Crecchio, Stotzky 2001; Meier et al. 2001). Miera hydrofóbonosti polutantu je teda jednou zo základných chemických charakteristík, ktorá ovplyvňuje jeho sorpciu. Sorpcia polutantov je navyše výrazne ovplyvnená množstvom vody v pôdnych systémoch. Voda sa pomerne pevne viaže na anorganické materiály a v tejto väzbe jej môžu v nasýtených systémoch konkurovať len relatívne polárne organické molekuly. Preto je aj pre polutanty s nižšou polaritou reaktívnou frakciou predovšetkým organická frakcia. Vo vodou nenasýtených pôdnych systémoch sa však sorpcia na minerálne povrchy alebo polárne oblasti organických látok môže stať dominantou aj pre nepolárne polutanty (Pennell et al. 1992). Vo všeobecnosti sorpcia polutantov redukuje rýchlosť biodegradácie znížením ich bioprístupnosti pre mikroorganizmy (Knezovich et al. 1987; Burgos et al. 1999). Napriek tomu bol zaznamenaný aj pozitívny vplyv sorpcie na biodegradáciu v prípade, keď sorpcia polutantu znížila jeho toxické účinky na mikroorganizmy (Earhardt, Rehm 1985). Bioprístupnosť sorbovaného polutantu je však závislá aj od druhu organizmu s biodegradačnou aktivitou (Guerin, Boyd 1997). Sorpcia na organickú frakciu má na biodegradáciu rôzny vplyv. Napríklad humínové látky majú na degradáciu polutantov aktivačný (Piccolo et al. 2001), ale aj inhibičný účinok (Thiele et al. 2002). Vplyv organickej frakcie závisí predovšetkým na sile vzniknutej väzby (polutant-organická frakcia), chemickej štruktúre organických látok, obsahu funkčných skupín a molekulovej hmotnosti. S rastúcim obsahom rozpustných organických látok v prítomnosti mikroorganizmov navyše rastie aj mobilita polutantu v prostredí (Kim et al. 2003; Kögel-Knabner, Totsche 1998). 116
3 Bioprístupnosť organických polutantov v prítomnosti ílových minerálov je závislá od pomeru ílu a roztoku, typu ílu a chemizmu polutantu. Sorpcia polutantov na expandovateľné ílové minerály znižuje bioprístupnosť organického kontaminantu (Magadliniuk et al. 1995), čo je spôsobené jeho migráciou do vnútorných vrstiev minerálu (Knaebel et al. 1996). Polutanty, ktroré majú záporný náboj alebo ich vedľajšie reťazce a veľkosť molekuly zabraňuje ich migrácii do vnútorných vrstiev, prítomnosť ílov neovplyvňuje. MIKROPÓROVÁ EXKLÚZIA Mikropórová exklúzia vzniká pri migrácii polutantov do oblasti mikropórov, ktoré fyzikálne znemožňujú prístup pre polutant degradujúce mikroorganizmy (Barra et al. 1998). Mikroorganizmy sú v pôdach distribuované medzi pevnou a kvapalnou fázou ako voľné alebo viazané na pevné substráty. Vo všeobecnosti nemôžu preniknúť do vnútorných oblastí pôdnych zŕn a mikropórov s priemerom menším ako 0,2 μm (Hassink et al. 1993). Polutanty sorbované v mikropóroch týchto rozmerov sa musia najskôr desorbovať a difundovať von z pórov. Mechanizmus transportu polutantu difúziou z mikropórov má určujúci vplyv na kinetiku biodegaradácie polutantu a je v poróznych systémoch limitujúcim faktorom biodegradácie. Ďalším možným mechanizmom degradácie takto viazaného polutantu je jeho degradácia exoenzýmom (Bertrand et al. 1993). Tento mechanizmus ale vo všeobecnosti nie je pre mikroorganizmus energeticky výhodný. Exoenzým musí navyše difundovať do póru a stéricky vhodne reagovať so substrátom. Metabolity reakcie musia následne difundovať von z pórov, aby boli prijaté bunkou mikroorganizmu (Hossain, Corapcioqlu 1996). ROZPUSTNOSŤ POLUTANTOV A JEJ VPLYV NA BIOPRÍSTUPNOSŤ V prípade polutantu s nízkou rozpustnosťou vo vode umožňuje afinita mikroorganizmov k hydrofóbnym povrchom naviazať sa na hydrofóbny substrát (Jones et al. 1991; Bunt et al. 1993) a priamo polutant absorbovať. Reverzibilné viazanie mikroorganizmu eliminuje problém obmedzeného prechodu pevnej fázy substrátu do roztoku. Mikroorganizmy sa na povrchy viažu špecifickým alebo nešpecifickým mechanizmami (Dufrěne et al. 1996), na čo majú vplyv najmä (1) elektrostatické a Van der Waalsove interakcie (2) a interakcie stérického charakteru medzi makromolekulami bunkového povrchu a substrátom (Rijnaarts et al. 1994). Veľkosť kontaktného povrchu substrátu s mikroorganizmom je faktor obmedzujúci túto stratégiu. Len minimálna časť bunkového povrchu je v priamom kontakte s adsorbovaným polutantom. Priamy príjem polutantu z povrchu pevnej fázy teda nie je dominantným mechanizmom. Bioprístupnosť polutantu vyžaduje, aby bol substrát rozpustený vo vodnej fáze obklopujúcej 117
4 bunku mikroorganizmu. V závislosti od chemicko-fyzikálnych vlastností systému, zahrňuje tento prenos desorpciu z pevnej fázy alebo oddelenie od fázy nevodných roztokov, ktorá môže byť zvýšená pôsobením biosurfaktantov. Biosurfaktanty môžu ovplyvniť biodegradáciu hydrofóbnych látok viacerými spôsobmi. Napríklad môžu zvýšiť ich bioprístupnosť prostredníctvom rozpúšťania molekúl do vodnej fázy (Bai et al. 1997), čo môže viesť k zvýšenej intenzite degradácie (Rahman et al. 2003; Uysal et Tűrkman 2005). Zhang et Miller (1994) napriek tomu uvádzajú, že biodegradácia sa po prídavku biosurfaktantu znížila. Účinok biosurfaktantov je teda pravdepodobne oveľa komplikovanejší, ako je ich vplyv na rozpustnosť hydrofóbnej látky. Môžu napríklad ovplyvniť hydrofóbnosť bunkového povrchu a tým väzbu bunky na substráty (Bai et al. 1997, Al-Tahhan et al. 2000). Je teda naďalej otázne, či rozpustenie polutantu skutočne musí predchádzať príjmu látky a jej biodegradáciu. ZÁVER V práci sme zhrnuli niektoré faktory ovplyvňujúce bioprístupnosť organických polutantov v pôdnych systémoch v súvislosti s možnosťou ich biodegradácie. Jednotlivé zložky pôdneho systému (ílové minerály, organické látky, mikroorganizmy, vodné a nevodné roztoky) a ich vlastnosti (napr. porozita, chemické zloženie) ovplyvňujú biodegradáciu polutantu mikroorganizmami v prírodných podmienkach rôznymi spôsobmi. POĎAKOVANIE Práca bola finančne podporená grantovými úlohami VEGA 1/3462/06 a VEGA 1/4361/07. LITERATÚRA Al-Tahhan R. A., Sandrin T. R., Bodour A. A., Maier R. M Rhamnolipidinduced removal lipopolysaccharide from Pseudomonas aeruginosa: Effect on cell surface properties and interaction with hydrophobic substrates. Appl. Environ. Microbiol. 66, pp Bai G., Brusseau M. L., Miller R. M Biosurfactant-enhanced removal of residual hydrocarbon from soil. J. Contam. Hydrol. 25, pp Barra M. T., Arias M., Guérif J Effect of iron and organic matter on porosity and structural stability of soil aggregates. Soil and Tillage Research 46, pp Berglöf T., Dung T.V., Kylin H., Nilsson I Carbendazim sorption-desorption in Vietnamese soils. Chemosphere 48, pp Bertrand J. C., Bianchi M., Mallah M. A., Acqaviva M., Mille G Hydrocarbon biodegradation and hydrocarbonoclastic bacterial communities composition grown 118
5 in seawater as a function of sodium chloride concentration. J. Exp. Mar. Biol. Ecol. 168, pp Bunt C. R., Jones D. S., Tucker I. G The effects of ph, ionic strength and organic phase on the bacterial adhesion to hydrocarobns (BATH) test. Int. J. Pharm. 99, pp Burgos W. D., Berry D. F., Bhandari A., Novak J. T Impact of soil-chemical interactions on the bioavailability of naphthalene and 1-naphthol. Water. Res. 33, pp Crecchio C., Stotzky G Biodegradation and insecticidal activity of the toxin from Bacillus thuringiensis subsp. kurstaki bound on complexes of montmorillonitehumic acids-al hydroxypolymers. Soil. Biol. Biochem. 33, pp Dufrěne Y. F., Boonaert C. J. P., Roxhet P. G Adhesion of Azospirillium brasilense: Role of proteins et the cell support interface. Colloids and Surfaces B: Biointerfaces 7, pp Ehrhardt H. M., Rehm H. J Phenol degradation by microorganisms adsorbed on activated carbon. Appl. Microbiol. Biotechnol. 21, pp Guerin W. F., Boyd S. A Biovailability of naphthalene associated with natural and synthetic sorbents. Water. Res. 31, pp Hassink J., Bouwman L. A., Zwart K. B., Brussaard L Relationship between habitable pore space, soil biota and mineralization rates in grassland soils. Soil. Biol. Biochem. 25, pp Hossain A., Corapcioqlu M. Y Modeling primary substrate controlled biotransformation and transport of halogenated aliphatics in porous media. Transport Porous Media 24, pp Iraqi S. M. U., Iraqi E Sorption of the pesticide endosulphan on two Indian soils. J. Colloid. Interface. Sci., 224, pp Jones D. S., Gorman S. P., McCafferty D. F., Woolfson A. D The effects of three non-antibiotic, antibacterial agents on the surface hydrophobicity of certain microorganisms evaluated by different methods. J. Appl. Bacteriol. 71, pp Kim S. B., Corapcioglu M. Y., Kim D. J Effect of dissolved organic matter and bacteria on contaminant transport in riverbank filtration. J. Contam. Hydrol. 66, pp Knaebel D. B., Federle T. W., McAwoy D. C., Vestal J. R Microbial mineralization of organic compounds in an acidic agricultural soil: effect of preadsorption to various soil contents. Environ. Toxicol. Chem. 15, pp Knezovich J. P., Harrison F. L., Wilhelm R. G The bioavailability of sedimentsorbed organic chemicals: A review. Water Air Soil Pollut. 32, pp Kögel-Knabner I., Totsche K. U Influence of dissolved and colloidal phase humic substances on the transport of the hydrophobic organic contaminants. Physics and Chemistry of the Earth 23, pp Magdaliniuk S., Block J. C., Leyval C., Bottero J. Y., Villemin G., Babut M Biodegradation of naphthalene in monmorillonite/polyacrimade suspension. Water. Sci. Tech. 31, pp Meier L. P., Nueesch R., Madsen F. T Organic pillared clays. J. Colloid Interface Sci. 238, pp
6 Mobed J. J., Hemminqsen S. L., Autry J. L., Mcqown L. B Fluorescence characterization of IHSS humic substances: Total luminiscence spectra with absorbance correction. Environ. Sci. Tech. 30, pp Pennel K. D., Rhue R. D., Rao P. S. C., Johnston C. T Vapor-phase sorption of p-xylene and water on soils and clay minerals. Environ. Sci. Tech Pettersson C., Ephraim J., Allard B On the composition and properties of humic substances isoltaed from deep groundwater and surface waters. Org. Geochem. 21, pp Piccolo A., Conte P., Cozzolino A., Paci M Combined effects of an oxidative enzyme and dissolved humic substances on 13C-labelled 2,4-D herbicide as revealed by high-resolution 13C NMR spectroscopy. J. Ind. Microbiol. Biotechnol. 26, pp Rahman K. S. M., Rahman T. J., Kourkoutas Y., Petsas I., Marchant R., Banat I. M Enhanced bioremediation of n-alkane in petroleum sludge using bacterial consortium amended with rhamnolipid and micronutrients. Bioresource Technology 90, pp Rijnaarts H. H. M., Norde W., Bouwer E. J., Lyklema J., Zehnder A. J. B Reversibility and mechanism of bacterial adhesion. Colloid and Surfaces B: Biointerfaces 4, pp Shenn Y. H Sorption of natural dissolved organic matter on soil. Chemosphere 38, pp Spark K. M., Swift R. S Effect of soil composition and dissolved organic matter on pesticide sorption. Sci. Total. Environ. 298, pp Thiele S., Fernandes E., Bollag J. M Enzymatic transformation and binding labeled 2,4,6-trinitrotoluene to humic substances during an anaerobic/aerobic incubation. J. Environ. Qual. 31, pp Uysal A., Tűrkman A Effect of biosurfactant on 2,4-dichlorphenol biodegradation in an activated sludge bioreactor. Process. Biochem. 40, pp Zhang Y., Miller R. M Effect of Pseudomonas rhamnolipid biosurfactant on cell hydrophobicity and biodegradation of octadecane. Appl. Environ. Microbiol. 60, pp
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