Biological Pretreatment Process Research Articles

Performances of full-scale biological nitrogen pre-treatment process for drinking water source: Seasonal variations and microbial community

This study investigated the performance of the full-scale unit over a two-year period to enhance nitrification efficiency and provide operational strategies. Results indicated that raw water quality from Donggan River was notably influenced by seasonal variations, particularly during dry and wet seasons, impacting the nitrification efficiency of the biological pretreatment process. Factors such as influent concentrations of ammonia and total Kjeldahl nitrogen were found to have significant effects on nitrification, with temperature and conductivity also showing correlations. The specific rate of ammonia removal was calculated to be approximately 0.1 kg-N/m3/d under the existing operational setup. Moreover, elevating dissolved oxygen levels above 4 mg/L was proposed to potentially boost ammonia oxidation based on findings from experiments conducted in lab-scale bioreactors. In times of increased influent ammonia levels, the elimination of about 1–3 mg-N/L of total nitrogen signified the activation of denitrification processes. This observation was corroborated by results from next-generation sequencing techniques, verifying the existence of denitrifying microorganisms. The real-time PCR analysis results indicated that the abundance of comammox amoA gene was comparable with the abundance of the AOB amoA gene, indicating the presence of comammox Nitrospira and their potential role on nitrification in the system.

Efficient production of cellooligosaccharides and xylooligosaccharides by combined biological pretreatment and enzymatic hydrolysis process

Efficient production of cellooligosaccharides and xylooligosaccharides by combined biological pretreatment and enzymatic hydrolysis process

Exploring the cellulolytic and hemicellulolytic activities of manganese peroxidase for lignocellulose deconstruction

BackgroundA cost-effective pretreatment and saccharification process is a necessary prerequisite for utilizing lignocellulosic biomass (LCB) in biofuel and biomaterials production. Utilizing a multifunctional enzyme with both pretreatment and saccharification functions in a single step for simultaneous biological pretreatment and saccharification process (SPS) will be a green method of low cost and high efficiency. Manganese peroxidase (MnP, EC 1.11.1.13), a well-known lignin-degrading peroxidase, is generally preferred for the biological pretreatment of biomass. However, exploring the role and performance of MnP in LCB conversion will promote the application of MnP for lignocellulose-based biorefineries.ResultsIn this study, we explored the ability of an MnP from Moniliophthora roreri, MrMnP, in LCB degradation. With Mn2+ and H2O2, MrMnP decomposed 5.0 g/L carboxymethyl cellulose to 0.14 mM of reducing sugar with a conversion yield of 5.0 mg/g, including 40 μM cellobiose, 70 μM cellotriose, 20 μM cellotetraose, and 10 μM cellohexaose, and degraded 1.0 g/L mannohexaose to 0.33 μM mannose, 4.08 μM mannotriose, and 4.35 μM mannopentaose. Meanwhile, MrMnP decomposed 5.0 g/L lichenan to 0.85 mM of reducing sugar with a conversion yield of 30.6 mg/g, including 10 μM cellotriose, 20 μM cellotetraose, and 80 μM cellohexose independently of Mn2+ and H2O2. Moreover, the versatility of MrMnP in LCB deconstruction was further verified by decomposing locust bean gum and wheat bran into reducing sugars with a conversion yield of 54.4 mg/g and 29.5 mg/g, respectively, including oligosaccharides such as di- and tri-saccharides. The catalytic mechanism underlying MrMnP degraded lignocellulose was proposed as that with H2O2, MrMnP oxidizes Mn2+ to Mn3+. Subsequently, it forms a complex with malonate, facilitating the degradation of CMC and mannohexaose into reducing sugars. Without H2O2, MrMnP directly oxidizes malonate to hydroperoxyl acetic acid radical to form compound I, which then attacks the glucosidic bond of lichenan.ConclusionThis study identified a new function of MrMnP in the hydrolysis of cellulose and hemicellulose, suggesting that MrMnP exhibits its versatility in the pretreatment and saccharification of LCB. The results will lead to an in-depth understanding of biocatalytic saccharification and contribute to forming new enzymatic systems for using lignocellulose resources to produce sustainable and economically viable products and the long-term development of biorefinery, thereby increasing the productivity of LCB as a green resource.

The Summary of Nitritation Process in Mainstream Wastewater Treatment

The application of the mainstream partial nitritation/anammox (PN/A) process is promising due to the huge cost reduction compared to traditional biological nitrogen removal. However, the nitrite production rate (NPR) of a biological nitritation pre-treatment process is relatively lower than the nitrite consumption rate in a pure anammox reactor with a high nitrogen loading rate (NLR). Thus, the NPR is the rate-limiting step for operating the PN/A process with a higher NLR. Various studies have attempted to improve mainstream NPR. A comprehensive review of these processes is needed for the actual application of the PN/A process. This study focuses on: (1) various nitrite production processes that have emerged in recent years; (2) the main microbial species and characteristics involved in biological nitritation; (3) the existing problems and the N2O emission problem of these processes; and (4) a proposed novel and promising PN/A process facilitated with photocatalyst oxidation. This review is expected to provide references and a basis for the research on the nitritation step of the application of the mainstream PN/A process.

Recyclable choline chloride-based deep eutectic solvent pretreatment for accelerated enzymatic digestibility of Triarrhena lutarioriparia

Choline chloride (ChCl) is the most common hydrogen acceptor (HBA) used in deep eutectic solvents (DES) due to its low toxicity and biodegradability. In this study, seven types of ChCl-based DESs were used for the pretreatment of Triarrhena lutarioriparia (TL), and the residues after pretreatment were subjected to enzymatic hydrolysis. Characterization techniques, including X-ray diffraction (XRD), scanning electron microscopy (SEM), and Fourier transformed infrared (FTIR) spectroscopy were used to determine the physical and chemical properties of TL pretreated with DESs. The results demonstrated that ChCl-oxalic acid dihydrate (molar ratio of 2:1; CO2:1) exhibited a good pretreatment ability among the DESs types examined. The optimal TL pretreatment process with CO2:1 was obtained by uniform design (UD) experiments and comprised a solid-liquid ratio of 1:25, temperature of 110 ℃, and time of 5 h, which achieved an enzymatic digestibility of 83.71 %. Furthermore, the cellulose reservation, hemicelluloses removal, and lignin removal still remained over 86 %, 80 % and 67 %, respectively, after being recycled five times. Overall, the CO2:1 pretreatment process was effective for the component separation and enzymatic hydrolysis of TL and will promote the development of a green biological refinery pretreatment process.

Greener Pretreatment Approaches for the Valorisation of Natural Fibre Biomass into Bioproducts.

The utilization of lignocellulosic biomass in various applications has a promising potential as advanced technology progresses due to its renowned advantages as cheap and abundant feedstock. The main drawback in the utilization of this type of biomass is the essential requirement for the pretreatment process. The most common pretreatment process applied is chemical pretreatment. However, it is a non-eco-friendly process. Therefore, this review aims to bring into light several greener pretreatment processes as an alternative approach for the current chemical pretreatment. The main processes for each physical and biological pretreatment process are reviewed and highlighted. Additionally, recent advances in the effect of different non-chemical pretreatment approaches for the natural fibres are also critically discussed with a focus on bioproducts conversion.

Biological Characterization and Instrumental Analytical Comparison of Two Biorefining Pretreatments for Water Hyacinth (Eichhornia crassipes) Biomass Hydrolysis

Water hyacinth is a rapidly growing troublesome aquatic weed plant, which causes eutrophication in water bodies and irreversible damage to the ecological system. In this work, we have investigated the water hyacinth biomass (WHB) hydrolysis efficacy of dilute alkaline (DA) pretreatment followed by biological pretreatment with white-rot fungus Alternaria alternata strain AKJK-2. The effectiveness of the dilute alkaline (DA) and biological pretreatment process on WHB was confirmed by using X-ray Diffraction (XRD) and Fourier Transform Infrared Spectrophotometer (FTIR), and was further visualized by Scanning Electron Microscope (SEM) and Confocal Laser Scanning Microscopy (CLSM). XRD spectra showed the increase in the crystallinity of pretreated samples, attributed to the elimination of amorphous components as lignin and hemicellulose. FTIR peak analysis of pre-treated WHB showed substantial changes in the absorption of cellulose functional groups and the elimination of lignin signals. Scanning electron microscopy (SEM) images showed firm, compact, highly ordered, and rigid fibril structures without degradation in the untreated WHB sample, while the pretreated samples exhibited loose, dispersed, and distorted structures. XRD indices (Segal, Landis, and Faneite), and FTIR indices [Hydrogen bond intensity (HBI); Total crystallinity index (TCI); and Lateral order crystallinity (LOI)] results were similar to the aforementioned results, and also showed an increase in the crystallinity both in alkaline and biological pretreatments. Alkaline pretreated WHB, with these indices, also showed the highest crystallinity and a crystalline allomorphs mixture of cellulose I (native) and cellulose II. These results were further validated by the CLSM, wherein fluorescent signals were lost after the pretreatment of WHB over control. Overall, these findings showed the significant potential of integrated assessment tools with chemical and biological pretreatment for large-scale utilization and bioconversion of this potential aquatic weed for bioenergy production.

Biological pretreatment of corn straw for enhancing degradation efficiency and biogas production

ABSTRACT In order to explore the effect of pretreatment on corn straw degradation and biogas production, corn straw was pretreated with mixed microbes and composting at 30°C for 14 days. The characteristics of material were measured and analyzed in the pretreatment process. Then, the pretreated and untreated corn straw was digested by anaerobic fermentation. Gas production and methane content of corn straw were analyzed. The results showed that the biological pretreatment process with mixed microbes could accelerate the degradation rate of straw and increase the degradation efficiency of lignin. The pH value of material was more stable, and the content of organic matter in the material was higher in the pretreatment process of corn straw with mixed microbes. The Scanning Electron Microscope (SEM) images showed that the structure of the lignocellulose was changed by mixed microbes, increasing the exposed area of cellulose and hemicellulose, which was beneficial to improve the utilization efficiency of straw. The degradation rates of hemicellulose, cellulose and lignin were 44.4%, 34.9% and 39.2%, respectively, after the pretreatment process with mixed microbes. Pretreatment was more helpful to increase the methane content in the anaerobic fermentation process of corn straw pretreated with mixed microbes, and could also shorten the fermentation period.

Enzymatic hydrolysis of biologically pretreated sorghum husk for bioethanol production

Biological pretreatment of lignocellulosic biomass is considered to be energy-efficient and cost-effective. In the present study, sorghum husk was biologically pretreated with a white-rot fungus Phanerochaete chrysosporium (MTCC 4955) under submerged static condition. Ligninolytic enzymes like lignin peroxidase (0.843 U/mL) and manganese peroxidase (0.389 U/mL) played an important role in the biological pretreatment of sorghum husk. Activities of different hydrolytic enzymes such as endoglucanase (57.25 U/mL), exoglucanase (4.76 U/mL), filter paperase (0.580 U/mL), glucoamylase (153.38 U/mL), and xylanase (88.14 U/mL) during biological pretreatment of sorghum husk by P. chrysosporium were evaluated. Enzymatic hydrolysis of untreated sorghum husk and biologically pretreated sorghum husk produced 20.07 and 103.0 mg/g reducing sugars, respectively. This result showed a significant increase in reducing sugar production in the biologically pretreated sorghum husk as compared to its untreated counterpart. Biologically pretreated sorghum husk hydrolysate was further fermented for 48 h using Saccharomyces cerevisiae (KCTC 7296), Pachysolen tannophilus (MTCC 1077), and their co-culture resulting in ethanol yields of 2.113, 1.095, and 2.348%, respectively. The surface characteristics of the substrate were evaluated after the delignification and hydrolysis, using FTIR, XRD, and SEM, confirming the effectiveness of the biological pretreatment process.

Analysis of biological pretreatment of rapeseed straw with white rot fungi for enzymatic hydrolysis

The effect of biological pretreatment on the rapeseed straw was evaluated by solid-state cultivation of white rot fungi Phanerochaete chrysosporium. P.chrysosporium degraded the lignin during the pretreatment, with enzymatic hydrolysis ratios increasing in the pretreated straw (3-fold after a 15 day pretreatment). The samples were identified by XRD, FTIR and SEM. X-ray analysis showed that pretreated samples had higher crystallinity than untreated samples (39.47% for a pretreated sample compared to 33.17% for untreated samples) and FTIR spectroscopy demonstrated that the content of lignocellulose also decreased during the biological pretreatment process. The surface characterization study showed morphological changes in pretreated samples. Moreover, the biological pretreatment slowed a plunge in hydrolysis rate during enzymatic hydrolysis.

Performance improvement of raw water pretreatment process with pre-inoculation biofilm: feasibility and limiting factors.

The initial formation of biofilm and the removal performance of pollutants in biological pretreatment process forpolluted raw water were limited due to the oligotrophic niche in raw water. In this study, the feasibility of using pre-inoculation biofilm formed under nutrients enhanced condition for polluted raw water treatment was analyzed in nine batch reactors. Results showed that the pollutants removal performance of biofilm was improved under nutrients enhanced conditions. Ammonia oxidation rate (AOR) was exponentially increased with the increasing in NH4+-N levels, and organic matter removal rate (ORR) was positively related to the initial total organic carbon (TOC) concentration. The biofilm formation and microbial diversity were further improved via adding more substrates, seeding river sediment and feeding effluent from a mature biofilm reactor. However, the biofilm formed under higher substrate conditions had higher half-saturation constant values (K S) to both NH4+-N and TOC, which decreased AOR and ORR values when it was used to treat polluted raw water. The reduction percentage of AOR and ORR showed logarithmic growth modes with the increase in K S values. Fortunately, improvement of nutrients flux via accelerating influent replacement could enhance the start-up performance effectively and decrease the operation risk introduced by the changes in substrate affinity.

A new biological process for short-chain fatty acid generation from waste activated sludge improved by Clostridiales enhancement.

Short-chain fatty acids (SCFAs), the carbon source of biological nutrient removal, can be produced by waste activated sludge (WAS) anaerobic fermentation. To get more SCFAs from sludge, most studies in literature focused on the mechanical process control or the structure of microbial community; little attention has been paid to the key microorganisms and their function related to SCFA generation. In this study, a different sludge pretreated method, i.e., pretreating sludge by proteinase K for 2days followed by pretreating at pH 10 for 4days, is reported, by which the proportion of Clostridiales was increased and SCFA generation was enhanced. First, the effects of different proteinase K concentrations and initial pH on sludge hydrolysis and SCFA generation were investigated. The optimal conditions showed the highest SCFA generation (352.91mg COD per gram of volatile suspended solids), which was 2.89-fold of the blank (un-pretreated). Further, the new biological pretreatment process led to the conversion of other SCFAs to acetic acid. Acetic acid accounted for 60.8% of total SCFAs with the new biological pretreatment process compared with 44.9% in the blank test. Then, the investigation on the key microorganisms related to SCFA production with 16S rRNA gene clone library and fluorescence in situ hybridization (FISH) indicated that there were much greater active Clostridiales when SCFAs were generated with the proteinase K and pH 10 pretreated sludge. Further, the mechanisms for the optimal conditions significantly enhancing SCFA generation were investigated. It was found that pretreating sludge by proteinase K and pH 10 caused the greatest key enzyme activities, organic consumption, and inhibition of methane generation. Graphical abstract A new biological process for short-chain fatty acid generation from waste activated sludge improved by Clostridiales enhancement.

Bio oil synthesis by coupling biological biomass pretreatment and catalytic hydroliquefaction process

The bio-oil synthesis from a mixture of wastes (7wt.% straw, 38wt.% wood, and 45wt.% grass) was carried out by direct liquefaction reaction using Raney Nickel as catalyst and tetralin as solvent. The green wastes were biologically degraded during 3 months. Longer the destructuration time; higher the yield into oil is. Biological pretreatment of green wastes promotes the liquefaction process. Among the components of degraded biomass, Humin, the major fraction (60-80wt.%) that was favored by the biological treatment, yields to a bio oil extremely energetic with a HHV close to biopetroleum (40MJ kg(-1)), contrariwise, Fulvic acids (2-12wt.%), the minor fraction is refractory to liquefaction reaction.

Evaluation of Biological Pretreatment of Rubberwood with White Rot Fungi for Enzymatic Hydrolysis.

The effects of biological pretreatment on the rubberwood (Hevea brasiliensis), was evaluated after cultivation of white rot fungi Ceriporiopsis subvermispora, Trametes versicolor, and a mixed culture of C. subvermispora and T. versicolor. The analysis of chemical compositions indicated that C. subvermispora had greater selectivity for lignin degradation with the highest lignin and hemicellulose loss at 45.06% and 42.08%, respectively, and lowest cellulose loss (9.50%) after 90 days among the tested samples. X-ray analysis showed that pretreated samples had a higher crystallinity than untreated samples. The sample pretreated by C. subvermispora presented the highest crystallinity of all the samples which might be caused by the selective degradation of amorphous components. Fourier transform infrared (FT-IR) spectroscopy demonstrated that the content of lignin and hemicellulose decreased during the biological pretreatment process. A study on hydrolysis of rubberwood treated with C. subvermispora, T. versicolor, and mixed culture for 90 days resulted in an increased sugar yield of about 27.67%, 16.23%, and 14.20%, respectively, as compared with untreated rubberwood (2.88%). The results obtained demonstrate that rubberwood is a potential raw material for industrial applications and white rot fungus C. subevermispora provides an effective method for improving the enzymatic hydrolysis of rubberwood.

Biodegradation of Lignocellulose by White-Rot Fungi: Structural Characterization of Water-Soluble Hemicelluloses

In the biological pretreatment process, white-rot fungi are mostly used to degrade lignin and carbohydrates in lignocellulosic biomass. In this study, water-soluble hemicelluloses were recovered from birch wood (Betula alnoides) decayed by white-rot fungi (Ganoderma lucidum C7016) for different durations up to 16 weeks. Accordingly, the dimethyl sulfoxide (DMSO)-soluble hemicelluloses were isolated from the untreated birch wood as a comparison. Results showed that the fungal-degraded polysaccharides were acidic hemicelluloses having a high content of uronic acids ranging from 20.6 to 22.5 %. Gel permeation chromatography analysis demonstrated that the recovered water-soluble hemicelluloses had a lower average molecular weight (M w, 15,990–27,560 g mol−1) than that of the DMSO-soluble hemicelluloses (M w , 33,960 g mol−1). Fourier transform infrared spectroscopy, scanning electron microscopy, one- and two-dimensional nuclear magnetic resonance spectroscopy also revealed significantly changes between those of fungal degraded and DMSO-soluble hemicelluloses. It was proposed that the hemicelluloses with low molecular weights were easily removed from wood by fungal degradation. This research revealed the changes of hemicelluloses in fungal degradation in the natural environment, which may enable the exploration of novel methods in bioconversion of lignocellulosic biomass for the production of biofuels and biopolymers, in addition to the development of new and better ways to protect wood from biodegradation by microorganisms.

The promoting effect of byproducts from Irpex lacteus on subsequent enzymatic hydrolysis of bio-pretreated cornstalks

BackgroundIrpex lacteus, a versatile lignin-degrading fungus with various extracellular enzymes, has been widely used for biological pretreatment. However, most studies have focused on the change of substrate structure after biological pretreatment, and the effect of these changes on the enzymatic hydrolysis, but the effect of byproducts from biological pretreatment process on subsequent enzymatic hydrolysis is not well understood.MethodsWe developed a biological pretreatment process with I. lacteus that can produce stimulatory byproducts that enhance the enzymatic hydrolysis of cornstalks.ResultsThe maximum hydrolysis yield of glucan (82%) was obtained after pretreatment for 28 days. The maximum reducing sugar yield decreased from 313.5 to 200.1 mg/g raw cornstalks after water-soluble byproducts of biological pretreatment were removed from pretreated cornstalks. The effect of byproducts on enzymatic hydrolysis was also investigated. We found that the hydrolysis efficiency of commercial cellulase preparation on cornstalks could be improved by water extracts from bio-pretreated cornstalks with hydrolytic enzyme activity and iron-reducing activity.ConclusionThe key finding suggested that byproducts from biological pretreatment play important roles in enhancing downstream hydrolysis, which might be attributable to hydrolytic enzymes and iron-reducing compounds produced by I. lacteus.

Biological Pretreatment of Lignocellulosic Substrates for Enhanced Delignification and Enzymatic Digestibility

Sheer enormity of lignocellulosics makes them potential feedstock for biofuel production but, their conversion into fermentable sugars is a major hurdle. They have to be pretreated physically, chemically, or biologically to be used by fermenting organisms for production of ethanol. Each lignocellulosic substrate is a complex mix of cellulose, hemicellulose and lignin, bound in a matrix. While cellulose and hemicellulose yield fermentable sugars, lignin is the most recalcitrant polymer, consisting of phenyl-propanoid units. Many microorganisms in nature are able to attack and degrade lignin, thus making access to cellulose easy. Such organisms are abundantly found in forest leaf litter/composts and especially include the wood rotting fungi, actinomycetes and bacteria. These microorganisms possess enzyme systems to attack, depolymerize and degrade the polymers in lignocellulosic substrates. Current pretreatment research is targeted towards developing processes which are mild, economical and environment friendly facilitating subsequent saccharification of cellulose and its fermentation to ethanol. Besides being the critical step, pretreatment is also cost intensive. Biological treatments with white rot fungi and Streptomyces have been studied for delignification of pulp, increasing digestibility of lignocellulosics for animal feed and for bioremediation of paper mill effluents. Such lignocellulolytic organisms can prove extremely useful in production of bioethanol when used for removal of lignin from lignocellulosic substrate and also for cellulase production. Our studies on treatment of hardwood and softwood residues with Streptomyces griseus isolated from leaf litter showed that it enhanced the mild alkaline solubilisation of lignins and also produced high levels of the cellulase complex when growing on wood substrates. Lignin loss (Klason lignin) observed was 10.5 and 23.5% in case of soft wood and hard wood, respectively. Thus, biological pretreatment process for lignocellulosic substrate using lignolytic organisms such as actinomycetes and white rot fungi can be developed for facilitating efficient enzymatic digestibility of cellulose.

Minimization of membrane organic fouling and haloacetic acids formation by controlling amino sugars and/or polysaccharide-like substances included in colloidal NOM

The objective of this study was to evaluate the effect of colloidal natural organic matter (NOM) on the disinfection by-products (DBPs) formation potentials and membrane organic fouling. From various analyses of a NOM-fouled ultrafiltration membrane surface, large amounts of colloidal NOM were found in the analyzed foulants. From FTIR spectra and pyrolysis analyses, colloidal NOMs were found to include amino sugars and polysaccharides, as indicated by N-acetyl groups in the FTIR spectra and from pyrolysis GC-mass analyses. Colloidal NOMs have two problematic aspects for membrane applications, which may induce bio-fouling due to their low biostabilities, and relatively high DBPs reactivities. The amino sugars and polysaccharides can be utilized by heterotrophic bacteria, and are identified as colloidal NOM with high fractions of biodegradable dissolved organic carbon (approximately 39%), so, their removal is expected to involve some form of biological pre-treatment process. Colloidal NOM also exhibit relatively high formation potentials of DBPs, especially haloacetic acids.

Biotreatment of an industrial waste oil condensate

The biotreatment of an industrial waste oil condensate has been investigated. The waste is an oily emulsion resulting from chemical processing and condensation of grease trap wastes and industrial waste oils. The oil consists of a complex mix of hydrocarbons with significant fuel oil and lube oil fractions. Currently this waste is disposed of by incineration. The feasibility of using a biological pretreatment process to remove a significant proportion of the hydrocarbons has been investigated. Enrichment cultures produced a stable bacterial consortium. Flask cultures of this enrichment culture were capable of rapid emulsification of the oil. Within 10 days, 40–50% of the oil waste was degraded. Degradation was monitored using gas chromatographic analysis with flame ionisation detector (GC-FID) and by assessment of microbial dehydrogenase activity using triphenyl tetrazolium chloride (TTC) dye reduction. The enrichment culture consisted of 9 component organisms, 7 Gram negative and one Gram positive organisms. Their degradative abilities in monoculture have been investigated. Degradation of the waste using monocultures was monitored using GC-FID analysis of the Pristane:C17 ratio in the waste. The degradation capability of each of the component organisms in pure culture was similar to that of the consortium.

Biotreatment of an industrial waste oil condensate

The biotreatment of an industrial waste oil condensate has been investigated. The waste is an oily emulsion resulting from chemical processing and condensation of grease trap wastes and industrial waste oils. The oil consists of a complex mix of hydrocarbons with significant fuel oil and lube oil fractions. Currently this waste is disposed of by incineration. The feasibility of using a biological pretreatment process to remove a significant proportion of the hydrocarbons has been investigated.Enrichment cultures produced a stable bacterial consortium. Flask cultures of this enrichment culture were capable of rapid emulsification of the oil. Within 10 days, 40–50% of the oil waste was degraded. Degradation was monitored using gas chromatographic analysis with flame ionisation detector (GC-FID) and by assessment of microbial dehydrogenase activity using triphenyl tetrazolium chloride (TTC) dye reduction. The enrichment culture consisted of 9 component organisms, 7 Gram negative and one Gram positive organisms. Their degradative abilities in monoculture have been investigated. Degradation of the waste using monocultures was monitored using GC-FID analysis of the Pristane:C17 ratio in the waste. The degradation capability of each of the component organisms in pure culture was similar to that of the consortium.