Biosurfactants are surface-active compounds produced by different microorganisms. The aim of this study was to introduce palm kernel cake (PKC) as a novel substrate for biosurfactant production using a potent bacterial strain under liquid state fermentation. This study was primarily based on the isolation and identification of biosurfactant-producing bacteria that could utilize palm kernel cake as a new major substrate. Potential bacterial strains were isolated from degraded PKC and screened for biosurfactant production with the help of the drop collapse assay and by analyzing the surface tension activity. From the screened isolates, a new strain, SM03, showed the best and most consistent results, and was therefore selected as the most potent biosurfactant-producing bacterial strain. The new strain was identified as Providencia alcalifaciens SM03 using the Gen III MicroPlate Biolog Microbial Identification System. The yield of the produced biosurfactant was 8.3 g/L.
A two-level fractional factorial design (FFD) was used to determine the effects of six factors, i.e. substrate (domestic wastewater sludge - DWS) and co-substrate concentration (wheat flour - WF), temperature, initial pH, inoculum size and agitation rate on the production of cellulase enzyme by Trichoderma harzianum in liquid state bioconversion. On statistical analysis of the results from the experimental studies, optimum process conditions were found to be temperature 32.5 degrees C, substrate concentration (DWS) 0.75% (w/w), co-substrate (WF) concentration 2% (w/w), initial pH 5, inoculum size 2% (v/w) and agitation 175 rpm. Analysis of variance (ANOVA) showed a high coefficient of determination (R2) of 0.975. Cellulase activity reached 10.2 FPU/ml at day 3 during the fermentation process which indicated about 1.5-fold increase in production compared to the cellulase activity obtained from the results of design of experiment (6.9 FPU/ml). Biodegradation of DWS was also evaluated to verify the efficiency of the bioconversion process as a waste management method.
Activated carbons derived from oil palm empty fruit bunches (EFB) were investigated to find the suitability of its application for removal of phenol in aqueous solution through adsorption process. Two types of activation namely; thermal activation at 300, 500 and 800 degrees C and physical activation at 150 degrees C (boiling treatment) were used for the production of the activated carbons. A control (untreated EFB) was used to compare the adsorption capacity of the activated carbons produced from these processes. The results indicated that the activated carbon derived at the temperature of 800 degrees C showed maximum absorption capacity in the aqueous solution of phenol. Batch adsorption studies showed an equilibrium time of 6 h for the activated carbon at 800 degrees C. It was observed that the adsorption capacity was higher at lower values of pH (2-3) and higher value of initial concentration of phenol (200-300 mg/L). The equilibrium data fitted better with the Freundlich adsorption isotherm compared to the Langmuir. Kinetic studies of phenol adsorption onto activated carbons were also studied to evaluate the adsorption rate. The estimated cost for production of activated carbon from EFB was shown in lower price (USD 0.50/kg of activated carbon) compared the activated carbon from other sources and processes.
The study was attempted to produce activated carbons from palm oil mill effluent (POME) sludge. The adsorption capacity of the activated carbons produced was evaluated in aqueous solution of phenol. Two types of activation were followed, namely, thermal activation at 300, 500 and 800 degrees C, and physical activation at 15 degrees C (boiling treatment). A control (raw POME sludge) was used to compare the adsorption capacity of the activated carbons produced. The results indicated that the activation temperature of 800 degrees C showed maximum absorption capacity by the activated carbon (POME 800) in aqueous solution of phenol. Batch adsorption studies showed an equilibrium time of 6 h for the activated carbon of POME 800. It was observed that the adsorption capacity was higher at lower values of pH (2-3) and higher value of initial concentration of phenol (200-300 mg/L). The equilibrium data were fitted by the Langmuir and Freundlich adsorption isotherms. The adsorption of phenol onto the activated carbon POME 800 was studied in terms of pseudo- first and second order kinetics to predict the rate constant and equilibrium capacity with the effect of initial phenol concentrations. The rate of adsorption was found to be better correlation for the pseudo-second order kinetics compared to the first order kinetics.
In this study, biodiesel was produced from sludge palm oil (SPO) using tolune-4-sulfonic monohydrate acid (PTSA) as an acid catalyst in different dosages in the presence of methanol to convert free fatty acid (FFA) to fatty acid methyl ester (FAME), followed by a transesterification process using an alkaline catalyst. In the first step, acid catalyzed esterification reduced the high FFA content of SPO to less than 2% with the different dosages of PTSA. The optimum conditions for pretreatment process by esterification were 0.75% (w/w) dosage of PTSA to SPO, 10:1 M ratio, 60 °C temperature, 60 min reaction time and 400 rpm stirrer speed. The highest yield of biodiesel after transesterification and purification processes was 76.62% with 0.07% FFA and 96% ester content. The biodiesel produced was favorable as compared to EN 14214 and ASTM 6751 standard. This study shows a potential exploitation of SPO as a new feedstock for the production of biodiesel.
This study enhanced the production of thermostable organic solvent-tolerant (TS-OST) lipase by locally isolated thermotolerant Rhizopus sp. strain using solid-state fermentation (SSF) of palm kernel cake (PKC). The optimum conditions were achieved using a series of statistical approaches. The cultivation parameters, which include fermentation time, moisture content, temperature, pH, inoculum size, various carbon and nitrogen sources, as well as other supplements, were initially screened by the definitive screening design, and one-factor-at-a-time using PKC as the basal medium. Three significant factors (olive oil concentration, pH, and inoculum size) were further optimized using face-centred central composite design. The results indicated a successful and significant improvement of lipase activity by almost two-fold compared to the initial screening production. The findings showed that the optimal conditions were 2% (v/w) inoculum size, 2% (v/w) olive oil, 0.6% (w/w) peptone, 2% (v/w) ethanol, 70% moisture content at initial pH 10.0 and 45 °C within 72 h of fermentation. Process optimization resulted in maximum lipase activity of 58.63 U/gram dry solids (gds). The analysis of variance showed that the statistical model was significant (p value <0.0001) and reliable with a high value of R2 (0.98) and adjusted R2 (0.96). This indicates a better correlation between the actual and predicted responses of lipase production. By considering this study, the low-cost PKC through SSF appears to be promising in the utilization of agro-industrial waste for TS-OST lipase production. This is because satisfactory enzyme activity could be attained that promises industrial applications.
A pilot-scale production of lipase using palm oil mill effluent (POME) as a fermentation basal medium was carried out, and parameters for immobilization of the produced lipase were optimized. Lipase production in a 300-L bioreactor was performed using two proposed strategies, constant power per volume (P/V) and constant tip speed. Moreover, lipase immobilization on different materials was also investigated. Lipase production was performed using liquid-state bioconversion of POME as the medium and Candida cylindracea as the inoculum. The fermentation medium was composed of 1% total suspended solids (TSS) of POME, 0.5% (w/v) peptone, 0.7% (v/v) Tween-80, and 2.2% inoculum. The medium composition was decided on the basis of the medium optimization results of a previous study. The fermentation was carried out for 48 h at 30°C and pH 6. The maximum lipase production was 5.72U/mL and 21.34 U/mL, obtained from the scale-up strategies of constant tip speed and P/V, respectively. Four accessible support materials were screened for their potential use in immobilization. The most suitable support material was found to be activated carbon, with a maximum immobilization of 94%.
A laboratory-scale study of bioconversion of local lignocellulosic material, oil palm biomass (OPB) was conducted by evaluating the enzyme production through microbial treatment in solid state bioconversion (SSB). OPB in the form of empty fruit bunches (EFB) was used as a solid substrate and treated with the white-rot fungus, Phanerochaete chrysosporium, to produce ligninase. The results showed that the highest ligninase activity of 400.27 U/liter was obtained at day 12 of fermentation. While the optimum study indicated the enzyme production of 1472.8 U/liter with moisture content of 50%, 578.7 U/liter with 10% v/w of inoculum size, and 721.8 U/liter with co-substrate concentration of 1% (w/w) at days 9, 9 and 12 of fungal treatment, respectively. The parameters glucosamine and reducing sugar were observed to evaluate the growth and substrate utilization in the experiment.
A laboratory-scale study was undertaken to evaluate the liquid state bioconversion (LSB) in terms of biodegradation of microbially treated domestic wastewater sludge (biosolids) as well as its kinetics. The potential fungal strains and process factors developed from previous studies were used throughout the study. The results presented in this study showed that an effective biodegradation occurred with the biosolids (sludge cake) accumulated. The maximum biosolids (sludge cake) accumulated (93.8 g/kg of liquid sludge) enriched with the biomass protein (30.2 g/kg of dry biosolids), was achieved which improved the effluent quality by enhancing the removal of chemical oxygen demand (COD), reducing sugar (RS), soluble protein (SP), total dissolved solids (TDS), and total suspended solids (TSS). The higher reduction of specific resistance to filtration (SRF) was observed during bioconversion process. The kinetics results showed that the experimental data were better fitted for the biodegradation efficiency, and biosolids accumulation and biodegradation rate.
Bioconversion of higher strength of domestic wastewater biosolids (sludge) (4% w/w of TSS) by mixed fungal culture of Aspergillus niger and Penicillium corylophilum was studied in a laboratory. The effect of potential mixed fungi on domestic wastewater sludge accelerated the liquid state bioconversion (LSB) process. The highest production of dry sludge cake (biosolids) was enriched with fungal biomass to about 85.66 g/kg containing 25.23 g/kg of protein after 8 days of treatment. The results presented in this study revealed that the reduction of chemical oxygen demand (COD), total suspended solid (TSS), and specific resistance to filtration (SRF) of treated sludge were highly influenced by the fungal culture as compared to control (uninnoculated). The maximum removal rates in treated sludge (biosolids) supernatant recorded were 92% of COD and 98.8% of TSS. Lower SRF (1.08 x 10(12) m/kg) was perceived in microbially treated sludge after 6 days of fermentation. The observed parameters were highly influenced after 8 days of treatment. The influence of pH was also studied and presented in the paper.
This study was undertaken to screen the filamentous fungi isolated from its relevant habitats(wastewater, sewage sludge and sludge cake) for the bioconversion of domestic wastewater sludge. A total of 35 fungal strains were tested against wastewater sludge (total suspended solids, TSS 1%-5% w/w) to evaluate its potentiality for enhancing the biodegradability and dewaterability using liquid state bioconversion(LSB) process. The strains were divided into five groups i.e. Penicillium, Aspergillus, Trichoderma, Basidiomycete and Miscellaneous, respectively. The strains WWZP1003, SCahmA103, SCahmT105 and PC-9 among their respective groups of Penicillium, Aspergillus, Trichoderma and Basidiomycete played potential roles in terms of separation (formation of pellets/flocs/filaments), biodegradation(removal of COD) and filtration (filterability) of treated domestic wastewater sludge. The Miscellaneous group was not considered due to its unsatisfactory results as compared to the other groups. The pH value was also influenced by the microbial treatment during fermentation process. The filterability of treated sludge was improved by fungal treatment, and lowest filtration time was recorded for the strain WWZP1003 and SCahmA103 of Penicillium and Aspergillus groups respectively compared with other strains.
The biosolids accumulation and biodegradation of domestic wastewater treatment plant (DWTP) sludge by filamentous fungi have been investigated in a batch fermenter. The filamentous fungi Aspergillus niger and Penicillium corylophilum isolated from wastewater and DWTP sludge was used to evaluate the treatment performance. The optimized mixed inoculum (A. niger and P. corylophilum) and developed process conditions (co-substrate and its concentration, temperature, initial pH, inoculum size, and aeration and agitation rate) were incorporated to accelerate the DWTP sludge treatment process. The results showed that microbial treatment of higher strength of DWTP sludge (4% w/w of TSS) was highly influenced by the liquid state bioconversion (LSB) process. In developed bioconversion processes, 93.8 g/kg of biosolids was enriched with fungal biomass protein of 30 g/kg. Enrichment of nutrients such as nitrogen (N), phosphorous (P), potassium (K) in biosolids was recorded in 6.2% (w/w), 3.1% (w/w) and 0.15% (w/w) from its initial values of 4.8% (w/w), 2.0% (w/w) and 0.08% (w/w) respectively after 10 days of fungal treatment. The biodegradation results revealed that 98.8% of TSS, 98.2% of TDS, 97.3% of turbidity, 80.2% of soluble protein, 98.8% of reducing sugar and 92.7% of COD in treated DWTP sludge supernatant were removed after 8 days of microbial treatment. The specific resistance to filtration (SRF) in treated sludge (1.4x10(12) m/kg) was decreased tremendously by the microbial treatment of DWTP sludge after 6 days of fermentation compared to untreated sample (85x10(12) m/kg).
The degradation potential and ligninolytic enzyme production of two isolated Panus tigrinus strains (M609RQY and M109RQY) were evaluated in this study. These strains were grown on three selected abundant agro-industrial wastes (rice straw; rice husk and cassava peel) under solid-state fermentation conditions. Degradation potential was determined by analyzing the chemical composition of the selected substrates before and after fermentation along with ligninolytic enzyme production. The strain M609RQY led to the highest lignin degradation of 40.81% on cassava peel, 11.25% on rice husk and 67.96% on rice straw. Both strains significantly increased the protein content of cassava peel. Rice husk stimulated maximum laccase (2556 U/L) and lignin peroxidase (24 U/L) production by the strains M109RQY and M609RQY, respectively. Furthermore, cassava peel stimulated maximum manganese-dependent peroxidase (141 U/L) production by the strain M109RQY. The de-lignified rice straw and the nutritionally-improved cassava peel could serve as potential animal feed supplements.
A green technology of biodiesel production focuses on the use of enzymes as the catalyst. In enzymatic biodiesel synthesis, suitable solvent system is very essential to reduce the inhibition effects of the solvent to the enzymes. This study produced ethanol-based biodiesel from a low-cost sludge palm oil (SPO) using locally-produced Candida cylindracea lipase from fermentation of palm oil mill effluent (POME) based medium. The optimum levels of ethanol-to-SPO molar ratio and enzyme loading were found to be 4:1 and 10 U/25 g of SPO respectively with 54.4% w/w SPO yield of biodiesel and 21.7% conversion of free fatty acid (FFA) into biodiesel. Addition of tert-butanol at 2:1 tert-butanol-to-SPO molar ratio into the ethanol-solvent system increased the yield of biodiesel to 71.6% w/w SPO and conversion of FFA into biodiesel to 28.8%. The SPO and ethanol have promising potential for the production of renewable biodiesel using enzymatic-catalyzed esterification and transesterification.
The pretreatment of empty fruit bunch (EFB) was conducted using an integrated system of IL and cellulases (IL-E), with simultaneous fermentation in one vessel. The cellulase mixture (PKC-Cel) was derived from Trichoderma reesei by solid-state fermentation. Choline acetate [Cho]OAc was utilized for the pretreatment due to its biocompatibility and biodegradability. The treated EFB and its hydrolysate were characterized by the Fourier transform infrared spectroscopy, scanning electron microscopy, and chemical analysis. The results showed that there were significant structural changes in EFB after the treatment in IL-E system. The sugar yield after enzymatic hydrolysis by the PKC-Cel was increased from 0.058 g/g of EFB in the crude sample (untreated) to 0.283 and 0.62 ± 06 g/g in IL-E system after 24 and 48 h of treatment, respectively. The EFB hydrolysate showed the eligibility for ethanol production without any supplements where ethanol yield was 0.275 g ethanol/g EFB in the presence of the IL, while lower yield obtained without IL-pretreatment. Moreover, it was demonstrated that furfural and phenolic compounds were not at the level of suppressing the fermentation process.
Lignocellulosic biomasses, exhibit resistance to enzymatic hydrolysis due to the presence of lignin and hemicellulose. Ionic liquids proved their applicability in lignin degradation, however, ionic liquid removal has to be performed to proceed to hydrolysis. Therefore, this study reports an in situ hydrolysis of empty fruit bunches (EFB) that combined an ionic liquid (IL) pretreatment and enzymatic hydrolysis. For enzyme production, palm kernel cake (PKC) was used as the primary media for microbial cellulase (PKC-Cel) from Trichoderma reesei (RUTC30). The obtained enzyme exhibited a promising stability in several ionic liquids. Among few, in choline acetate [Cho]OAc, PKC-Cel retained 63.16 % of the initial activity after 6 h and lost only 10 % of its activity in 10 % IL/buffer mixture. Upon the confirmation of the PKC-Cel stability, EFB was subjected to IL-pretreatment followed by hydrolysis in a single step without further removal of the IL. The findings revealed that choline acetate [Cho]OAc and choline butyrate [Cho]Bu were among the best ILs used in the study since 0.332 ± 0.05 g glucose/g and 0.565 ± 0.08 g total reducing sugar/g EFB were obtained after 24 h of enzymatic hydrolysis. Compared to the untreated EFB, the amount of reducing sugar obtained after enzymatic hydrolysis increased by three-fold in the case of [Cho]OAc and [Cho]Bu, two-fold with [EMIM]OAc and phosphate-based ILs whereas the lowest concentration was obtained in [TBPH]OAc. Pretreatment of EFB with [Cho]OAc and [Cho]Bu showed significant differences in the morphology of EFB samples when observed with SEM. Analysis of the lignin, hemicellulose and hemicellulose showed that the total lignin content from the raw EFB was reduced from 37.8 ± 0.6 to 25.81 ± 0.35 % (w/w) upon employment of [Cho]OAc in the compatible system. The PKC-Cel from T. reesei (RUTC30) exhibited promising characteristics that need to be investigated further towards a single-step process for bioethanol production.
Optimization of decolorization of methylene blue (MB) dye by lignin peroxidase (LiP) enzyme produced by white-rot fungus Phanerochaete chrysosporium using sewage treatment plant (STP) sludge as a major substrate was carried out in the laboratory. Optimization by the one-factor-at-a-time (OFAT) and statistical approach was carried out to determine the process conditions on optimum decolorization of MB dye using LiP enzyme in static mode. The OFAT method indicated that the optimum conditions for decolorization of MB dye (removal: 14-40%) was at temperature 55 degrees C, pH 5.0 with hydrogen peroxide (H(2)O(2)) concentration 4.0mM, MB dye concentration 20mg/L and LiP activity 0.487U/ml. The addition of veratryl alcohol to the reaction mixtures did not contribute any further increases in decolorization. The initial concentration of MB and the activity of LiP enzyme were further optimized using response surface methodology (RSM). The contour and surface plots suggested that the optimum initial concentration of MB and LiP activity predicted were 15mg/L and 0.687U/ml, respectively for the removal of 65%. The validation of the model showed that the decolorization process gave the higher removal of 90% in agitation mode compared to the static mode with 65% for 60min of incubation time by LiP enzyme.
A laboratory-scale study was carried out to produce lignin peroxidase (ligninase) by white rot fungus (Phanerochaete chrysosporium) using sewage-treatment-plant (STP) sludge as the major substrate. The optimization was done using full-factorial design (FFD) with agitation and aeration as the two parameters. Nine experiments indicated by the FFD were fermented in a stirred-tank bioreactor for 3 days. A second-order quadratic model was developed using the regression analysis of the experimental results with the linear, quadratic, and interaction effects of the parameters. Analysis of variance (ANOVA) showed a high coefficient of determination (R (2)) value of 0.972, thus indicating a satisfactory fit of the quadratic model with the experimental data. Using statistical analysis, the optimum aeration and agitation rates were determined to be 2.0 vvm and 200 rpm, respectively, with a maximum activity of 225 U l(-1) in the first 3 days of fermentation. The validation experiment showed the maximum activity of lignin peroxidase was 744 U l(-1) after 5 days of fermentation. The results for the tests of the stability of lignin peroxidase showed that the activity was more than 80% of the maximum for the first 12 h of incubation at an optimum pH of 5 and temperature of 55 degrees C.
The purpose of this study was to evaluate the feasibility of producing bioethanol from palm-oil mill effluent generated by the oil-palm industries through direct bioconversion process. The bioethanol production was carried out through the treatment of compatible mixed cultures such as Thrichoderma harzianum, Phanerochaete chrysosporium, Mucor hiemalis, and yeast, Saccharomyces cerevisiae. Simultaneous inoculation of T. harzianum and S. cerevisiae was found to be the mixed culture that yielded the highest ethanol production (4% v/v or 31.6 g/l). Statistical optimization was carried out to determine the operating conditions of the stirred-tank bioreactor for maximum bioethanol production by a two-level fractional factorial design with a single central point. The factors involved were oxygen saturation level (pO(2)%), temperature, and pH. A polynomial regression model was developed using the experimental data including the linear, quadratic, and interaction effects. Statistical analysis showed that the maximum ethanol production of 4.6% (v/v) or 36.3 g/l was achieved at a temperature of 32 degrees C, pH of 6, and pO(2) of 30%. The results of the model validation test under the developed optimum process conditions indicated that the maximum production was increased from 4.6% (v/v) to 6.5% (v/v) or 51.3 g/l with 89.1% chemical-oxygen-demand removal.