Optimisation of propionibacterial ECP production and the influence of propionibacteria on the UASB granulation process

Joubert, Hannarine (2000-12)

Thesis (MSc)--Stellenbosch University, 2000.

Thesis

ENGLISH ABSTRACT: The "classical" propionibacteria are used in a variety of natural dairy fermentations where they produce natural preservatives (propionic and acetic acids and bacteriocins) and large amounts of vitamin B12. The extracellular polysaccharide (ECP) producing ability of these bacteria also make them of special interest to the food and waste water management industries as the ECP has been illustrated to playa role in the initial granule formation in upflow anaerobic bioreactor systems. There is little known on the ECP production by propionibacteria and in this study different environmental conditions that influence ECP production were studied. Nineteen different Propionibacterium strains were examined in terms of ECP production and Propionibacterium strain 278 was identified as the best ECP producer. Further studies were only done on this strain because of its high ECP production and because it was originally isolated from an anaerobic digester. The influence of temperature, pH and sucrose concentration was determined through the measurement of ECP production and medium viscosity. It was found that more ECP was produced at temperatures lower than the optimum for growth with the optimum being between 22° and 25°C. Lower initial pH conditions of the growth medium (below pH 7.0) were found to inhibit ECP production and the influence when the initial pH values were between 7.0 and 8.5, was not significant. A higher carbon: nitrogen ratio, when 8% sucrose was added, was also found to enhance the ECP production. The upflow anaerobic sludge bed (UASB) bioreactor process depends on the upward movement of soluble matter through a blanket of active methanogenic granular sludge. The long start-up times as a result of the slow granulation process, as well as the need for a speedy replacement of granules once they have been washed out of the system, are limitations that restrict the general application of this excellent waste water treatment technology. Full exploitation of this biomass immobilisation technique can thus not be realised until the granule formation conditions are defined and optimised. The precise nature of the mechanisms involved in the formation of granules and the reason for their stability, is still not fully understood. It was hypothised by Britz et al. in 1999 that, through the implementation of environmental 'stress' conditions, a shift in the population dynamics of the anaerobic community can be obtained. This results in a concurrent increase in ECP formation that appears to enhance aggregate formation. In the second study it was found that, when 'stress' conditions were applied to already formed granules, the Gram-positive lactate-utilising acidogenic population gained an advantage and more propionic acid producing bacteria were present. The propionic and acetic acid concentrations were also found to increase, and concurrently, a decrease in the growth medium pH occurred. This confirms part of the granulation hypothesis that, when granules are 'stressed', the acidogenic population dynamics change and the lactate-utilising population responds to the gradual decrease in pH and the more acid-tolerant propionic acid producing bacteria gain a competitive advantage resulting in the increase in the propionic acid concentration. When propionibacteria were added to raw sludge during the granule production process, the granules were found to be more active than when nopropionibacteria had been added. This was probably due to the ECP formation by the propionibacteria that enhances the aggregation of the granules. Enhanced granulation was thus found in the batch systems with the fatty acids formed in correlation with the model for granulation. A good correlation was evident between the hypothesis and the experimental data and the hypothesis was partially verified in this study.

AFRIKAANSE OPSOMMING: Die "klassieke" propionibakterieë word in 'n verskeidenheid van natuurlike suiwel fermantasies gebruik waarin hulle verantwoordelik in vir die produksie van natuurlike voedsel preserveermiddels (propioonsuur, asynsuur en bakteriosiene) en groot hoeveelhede vitamiene B12. Die Ekstra Sellulêre Pollisakkaried (ESP) produserende eienskap van hierdie groep bakterieë maak hulle ook van belang in die voedsel en afvoerwater beheer industrieë, aangesien gevind is dat ESP 'n rol speel in die aanvanklike granule formasie in anaerobiese bioreaktor sisteme. Daar is nog baie min bekend oor die ESP produksie van propionibakterieë en in hierdie studie is verskeie omgewings faktore wat die ESP produksie beïnvloed, bestudeer. Negentien verskillende Propionibakterium stamme was bestudeer in terme van ESP produksie en Propionibakterium stam 278 was geïdentifiseer as die stam wat die meeste ESP produseer. Verdere studies was op hierdie stam gedoen na aanleiding van sy hoë ESP produksie en omdat dit oorspronklik uit 'n anaerobiese verteerder geisoleer is. Die invloed van termperatuur, pH en sukrose konsentrasie was bepaal deur die meting van die ESP produksie en die medium viskositeit. Dit was gevind dat meer ESP geproduseer was by temperature laer as die optimum vir groei, met die optimum temperatuur tussen 22° en 25°C. Dit is ook gevind dat laer aanvangs groei-medium pH (laer as pH 7.0), ESP produksie inhibeer. Die invloed van die aanvangs groei-medium pH tussen 7.0 en 8.5 was egter nie betekenisvol nie. Dit is ook gevind dat 'n hoër koolstof tot stikstof verhouding, verkry deur die byvoeging van 8% sukrose, die ESP produksie verhoog. Die "upflow anaerobic sludge blanket" (UASB) proses vind plaas as gevolg van die opwaarste beweging van opgeloste organiese materiaal deur 'n granule bed van aktiewe metanogeniese granulêre slyk. Die lang 'start-up' tye as gevolg van die stadige granulasie proses, en die nodigheid om 'n vinnige verplasing van granules te hê nadat dit uit die sisteem gewas is, is beperkings wat die algemene toepassing van hierdie fantastiese afvoerwater tegnologie, strem. Volle implementering van hierdie biomassa immobilisereings tegniek kan dus nie plaasvind voordat die granule formasie gedefinieer en geoptimiseer is nie. Die presiese eienskappe van die meganismes betrokke en die formasie van die granules en die rede vir hul stabiliteit word egter nog nie ten volle verstaan nie. Volgens 'n hipotese deur Britz et al. (1999), vind 'n verskuiwing in die populasie dinamika van die anaerobiese gemeenskap plaas tydens die implementasie van omgewings 'stress' toestande. Die resultaat is 'n verhoging in ESP produksie en 'n gevolglike verbetering in die granulasie proses. In die tweede studie was dit gevind dat, wanneer 'stress' toestande op die reeds gevormde granulasie toegepas word, die Gram-positiewe laktaat-benuttende asetogeniese populasie voordeel geniet en meer propioonsuur produserende bakterieë was teenwoordig. Die propioonsuur en asynsuur konsentrasies het ook verhoog en met 'n gevolglike daling in die groei-medium se pH. Dit bevestig 'n gedeelte van die hipotese dat, wanneer die granules onder 'stress' geplaas word, die asetogeniese populasie dinamika verander en die laktaat-benuttende populasie reageer tot die gedeeltelike afname in pH. Die meer suur-tolerante propioonsuur produserende bakterieë verkry 'n kompeterende voordeel en gevolglik is daar 'n verhoging in propioonsuur konsentrasie. Propionibakterieë was gevoeg by die onbehandelde slyk gedurende die granule produksie proses, en daar is gevind dat meer aktiewe granules gevorm word as andersins. Dit is moontlik as gevolg van die die ESP produksie van propionibakterieë wat die granulasie versnel het. Verbeterde granulasie was dus verkry in die sisteme waar propionibakterieë bygevoeg is. Vetsuur analises het gedui dat die gevormde vetsure ook in korrelasie was met die model van granulasie. Goeie korrelasie was dus verkry tussen die hipotese en die eksperimentele data en die hipotese is gedeeltelik bewys in hierdie studie.

Please refer to this item in SUNScholar by using the following persistent URL: http://hdl.handle.net/10019.1/51866
This item appears in the following collections: