It was found that with replenishment, powdered activated carbon (PAC) in the membrane bioreactor (MBR) would develop biologically activated carbon (BAC) which could enhance filtration performance of a conventional MBR. This paper addresses two issues (i) effect of PAC size on MBR (BAC) performance; and (ii) effect of sludge retention time (SRT) on the MBR performance with and without PAC. To interpret the trends, particle/floc size, concentration of mixed liquor suspended solid (MLSS), total organic carbon (TOC), short-term filtration properties and transmembrane pressure (TMP) versus time are measured. The results showed improved fouling control with fine, rather than coarse, PAC provided the flux did not exceed the deposition flux for the fine PAC. Without PAC, the longer SRT operation gave lower fouling at modest fluxes. With PAC addition, the shorter SRT gave better fouling control, possibly due to greater replenishment of the fresh PAC.
This study was to investigate the mineralization of wastewater containing methyl orange (MO) in integrated anaerobic-aerobic biofilm reactor with coconut fiber as bio-material. Different aeration periods (3h in phase 1 and 2; 3, 6 and 15 h in phase 3; 24 h in phase 4 and 5) in aerobic chamber were studied with different MO concentration 50, 100, 200, 200 and 300 mg/L as influent from phase 1-5. The color removals estimated from the standard curve of dye versus optical density at its maximum absorption wavelength were 97%, 96%, 97%, 97%, and 96% and COD removals were 75%, 72%, 63%, 81%, and 73% in phase 1-5, respectively. The MO decolorization and COD degradation followed first-order kinetic model and second-order kinetic model, respectively. GC-MS analysis indicated the symmetrical cleavage of azo bond and the reduction in aromatic peak ensured the partial mineralization of MO.
Microbial mannanases have become biotechnologically important in industry but their application is limited due to high production cost. In presents study, the extraction of mannanase from fermented Palm Kernel Cake (PKC) in the Solid State Fermentation (SSF) was optimized. Local isolate of Aspergillus terreus SUK-1 was grown on PKC in (SSF) using column bioreactor. The optimum condition were achieved after two washes of fermented PKC by adding of 10% glycerol (v/v) soaked for 10 h at the room temperature with solvent to ratio, 1:5 (w/v).
This study aims at developing a novel effluent removal management tool for septic sludge treatment plants (SSTP) using a clonal selection algorithm (CSA). The proposed CSA articulates the idea of utilizing an artificial immune system (AIS) to identify the behaviour of the SSTP, that is, using a sequence batch reactor (SBR) technology for treatment processes. The novelty of this study is the development of a predictive SSTP model for effluent discharge adopting the human immune system. Septic sludge from the individual septic tanks and package plants will be desuldged and treated in SSTP before discharging the wastewater into a waterway. The Borneo Island of Sarawak is selected as the case study. Currently, there are only two SSTPs in Sarawak, namely the Matang SSTP and the Sibu SSTP, and they are both using SBR technology. Monthly effluent discharges from 2007 to 2011 in the Matang SSTP are used in this study. Cross-validation is performed using data from the Sibu SSTP from April 2011 to July 2012. Both chemical oxygen demand (COD) and total suspended solids (TSS) in the effluent were analysed in this study. The model was validated and tested before forecasting the future effluent performance. The CSA-based SSTP model was simulated using MATLAB 7.10. The root mean square error (RMSE), mean absolute percentage error (MAPE), and correction coefficient (R) were used as performance indexes. In this study, it was found that the proposed prediction model was successful up to 84 months for the COD and 109 months for the TSS. In conclusion, the proposed CSA-based SSTP prediction model is indeed beneficial as an engineering tool to forecast the long-run performance of the SSTP and in turn, prevents infringement of future environmental balance in other towns in Sarawak.
The concept of de novo metabolic engineering through novel synthetic pathways offers new directions for multi-step enzymatic synthesis of complex molecules. This has been complemented by recent progress in performing enzymatic reactions using immobilized enzyme microreactors (IEMR). This work is concerned with the construction of de novo designed enzyme pathways in a microreactor synthesizing chiral molecules. An interesting compound, commonly used as the building block in several pharmaceutical syntheses, is a single diastereoisomer of 2-amino-1,3,4-butanetriol (ABT). This chiral amino alcohol can be synthesized from simple achiral substrates using two enzymes, transketolase (TK) and transaminase (TAm). Here we describe the development of an IEMR using His6-tagged TK and TAm immobilized onto Ni-NTA agarose beads and packed into tubes to enable multi-step enzyme reactions. The kinetic parameters of both enzymes were first determined using single IEMRs evaluated by a kinetic model developed for packed bed reactors. The Km(app) for both enzymes appeared to be flow rate dependent, while the turnover number kcat was reduced 3 fold compared to solution-phase TK and TAm reactions. For the multi-step enzyme reaction, single IEMRs were cascaded in series, whereby the first enzyme, TK, catalyzed a model reaction of lithium-hydroxypyruvate (HPA) and glycolaldehyde (GA) to L-erythrulose (ERY), and the second unit of the IEMR with immobilized TAm converted ERY into ABT using (S)-α-methylbenzylamine (MBA) as amine donor. With initial 60mM (HPA and GA each) and 6mM (MBA) substrate concentration mixture, the coupled reaction reached approximately 83% conversion in 20 min at the lowest flow rate. The ability to synthesize a chiral pharmaceutical intermediate, ABT in relatively short time proves this IEMR system as a powerful tool for construction and evaluation of de novo pathways as well as for determination of enzyme kinetics.
Volumetric mass transfer coefficient (kLa) is an important parameter in bioreactors handling viscous fermentations such as xanthan gum production, as it affects the reactor performance and productivity. Published literatures showed that adding an organic phase such as hydrocarbons or vegetable oil could increase the kLa. The present study opted for palm oil as the organic phase as it is plentiful in Malaysia. Experiments were carried out to study the effect of viscosity, gas holdup, and kLa on the xanthan solution with different palm oil fractions by varying the agitation rate and aeration rate in a 5 L bench-top bioreactor fitted with twin Rushton turbines. Results showed that 10% (v/v) of palm oil raised the kLa of xanthan solution by 1.5 to 3 folds with the highest kLa value of 84.44 h(-1). It was also found that palm oil increased the gas holdup and viscosity of the xanthan solution. The kLa values obtained as a function of power input, superficial gas velocity, and palm oil fraction were validated by two different empirical equations. Similarly, the gas holdup obtained as a function of power input and superficial gas velocity was validated by another empirical equation. All correlations were found to fit well with higher determination coefficients.
Nine aerobic cellulolytic bacterial cultures were obtained from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Culture (DSMZ) and the American Type Culture Collection (ATCC). The objectives of this study were to characterize the cellulolytic bacteria and to determine the optimum moisture ratio required for solid state fermentation (SSF) of palm kernel cake (PKC). The bacteria cultures were grown on reconstituted nutrient broth, incubated at 30°C and agitated at 200 rpm. Carboxymethyl cellulase, xylanase, and mannanase activities were determined using different substrates and after SSF of PKC. The SSF was conducted for 4 and 7 days with inoculum size of 10% (v/w) on different PKC concentration-to-moisture ratios: 1 : 0.2, 1 : 0.3, 1 : 0.4, and 1 : 0.5. Results showed that Bacillus amyloliquefaciens 1067 DSMZ, Bacillus megaterium 9885 ATCC, Paenibacillus curdlanolyticus 10248 DSMZ, and Paenibacillus polymyxa 842 ATCC produced higher enzyme activities as compared to other bacterial cultures grown on different substrates. The cultures mentioned above also produced higher enzyme activities when they were incubated under SSF using PKC as a substrate in different PKC-to-moisture ratios after 4 days of incubation, indicating that these cellulolytic bacteria can be used to degrade and improve the nutrient quality of PKC.
Lipase-catalyzed production of triethanolamine-based esterquat by esterification of oleic acid (OA) with triethanolamine (TEA) in n-hexane was performed in 2 L stirred-tank reactor. A set of experiments was designed by central composite design to process modeling and statistically evaluate the findings. Five independent process variables, including enzyme amount, reaction time, reaction temperature, substrates molar ratio of OA to TEA, and agitation speed, were studied under the given conditions designed by Design Expert software. Experimental data were examined for normality test before data processing stage and skewness and kurtosis indices were determined. The mathematical model developed was found to be adequate and statistically accurate to predict the optimum conversion of product. Response surface methodology with central composite design gave the best performance in this study, and the methodology as a whole has been proven to be adequate for the design and optimization of the enzymatic process.
The study is to identify the extraction of intracellular inulinase (exo- and endoinulinase) and invertase as well as optimization medium composition for maximum productions of intra- and extracellular enzymes from Aspergillus niger ATCC 20611. From two different methods for extraction of intracellular enzymes, ultrasonic method was found more effective. Response surface methodology (RSM) with a five-variable and three-level central composite design (CCD) was employed to optimize the medium composition. The effect of five main reaction parameters including sucrose, yeast extract, NaNO₃, Zn⁺², and Triton X-100 on the production of enzymes was analyzed. A modified quadratic model was fitted to the data with a coefficient of determination (R²) more than 0.90 for all responses. The intra-extracellular inulinase and invertase productions increased in the range from 16 to 8.4 times in the optimized medium (10% (w/v) sucrose, 2.5% (w/v) yeast extract, 2% (w/v) NaNO₃, 1.5 mM (v/v) Zn⁺², and 1% (v/v) Triton X-100) by RSM and from around 1.2 to 1.3 times greater than in the medium optimized by one-factor-at-a-time, respectively. The results of bioprocesses optimization can be useful in the scale-up fermentation and food industry.
The hybridoma 192 was used to produce a monoclonal antibody (MAb) against 17-hydroxyprogesterone (17-OHP), for possible use in screening for congenital adrenal hyperplasia (CAH). The factors influencing the MAb production were screened and optimized in a 2 L stirred bioreactor. The production was then scaled up to a 20 L bioreactor. All of the screened factors (aeration rate, stirring speed, dissolved oxygen concentration, pH, and temperature) were found to significantly affect production. Optimization using the response surface methodology identified the following optimal production conditions: 36.8°C, pH 7.4, stirring speed of 100 rpm, 30% dissolved oxygen concentration, and an aeration rate of 0.09 vvm. Under these conditions, the maximum viable cell density achieved was 1.34 ± 0.21 × 10(6) cells mL(-1) and the specific growth rate was 0.036 ± 0.004 h(-1) . The maximum MAb titer was 11.94 ± 4.81 μg mL(-1) with an average specific MAb production rate of 0.273 ± 0.135 pg cell(-1) h(-1) . A constant impeller tip speed criterion was used for the scale-up. The specific growth rate (0.040 h(-1) ) and the maximum viable cell density (1.89 × 10(6) cells mL(-1) ) at the larger scale were better than the values achieved at the small scale, but the MAb titer in the 20 L bioreactor was 18% lower than in the smaller bioreactor. A change in the culture environment from the static conditions of a T-flask to the stirred bioreactor culture did not affect the specificity of the MAb toward its antigen (17-OHP) and did not compromise the structural integrity of the MAb.
Biological removal of carbon, nitrogen and sulfur is drawing increasing research interest in search for an efficient and cost-effective wastewater treatment. While extensive work on separate removal of nitrogen and sulfur is well documented, investigation on simultaneous denitrifying sulfide removal has only been reported recently. Most of the work on denitrifying sulfide removal has been focusing on bioreactor performance, loading and operating conditions. Nonetheless, underlying principles elucidating the biochemical reactions and the mechanisms of the microbial degradation are yet to be established. In addition, unstable denitrifying sulfide removal which is a major operating problem that hinders practical application of the process, is yet to be resolved. This paper provides a review on the state-of-the-art development of simultaneous biological removal of sulfur, nitrogen and carbon. Research on bioreactor operation and performance, reactor configurations, mechanisms and modeling work including the use of mass balance analysis and artificial neural networks is delineated. An in-depth discussion on the microbial community and functional consortium is also provided. Challenges and future work on simultaneous biological removal of nitrogen-sulfur-carbon are also outlined.
To date, little information is known about the operation of the enhanced biological phosphorus removal (EBPR) process in tropical climates. Along with the global concerns on nutrient pollution and the increasing array of local regulatory requirements, the applicability and compliance accountability of the EBPR process for sewage treatment in tropical climates is being evaluated. A sequencing batch reactor (SBR) inoculated with seed sludge from a conventional activated sludge (CAS) process was successfully acclimatized to EBPR conditions at 28 °C after 13 days' operation. Enrichment of Candidatus Accumulibacter phosphatis in the SBR was confirmed through fluorescence in situ hybridization (FISH). The effects of operational pH and influent C:P ratio on EBPR were then investigated. At pH 7 or pH 8, phosphorus removal rates of the EBPR processes were relatively higher when operated at C:P ratio of 3 than C:P ratio of 10, with 0.019-0.020 and 0.011-0.012 g-P/g-MLVSS•day respectively. One-year operation of the 28 °C EBPR process at C:P ratio of 3 and pH 8 demonstrated stable phosphorus removal rate of 0.020 ± 0.003 g-P/g-MLVSS•day, corresponding to effluent with phosphorus concentration <0.5 mg/L. This study provides the first evidence on good EBPR activity at relatively high temperature, indicating its applicability in a tropical climate.
This study was conducted to determine the total phenolic (TPC) and total flavonoid content (TFC) as well as the antioxidant activity of 50% ethanolic extracts from different parts of Camellia sinensis (shoot, young and matured leaves). Comparison was also made between black (fermented) and green (unfermented) tea. For green tea, the results showed that the shoot contained significantly higher total phenolic content, followed by the young and matured leaves (p
Nipa sap or air nira is a sweet natural beverage obtained from a type of palm tree, Nypa fruticans.
It is readily and spontaneously fermented resulting in the development of alcoholic fermentation products. Objective of this study is to determine the volatile compounds (VOCs) responsible for the aroma in fresh and fermented nipa sap. The sap was left for natural fermentation at 30ºC for 63 days. VOCs of the sap were analysed using static headspace gas chromatography-mass spectrometry (GC-MS). Fresh nipa sap contained ethanol (83.43%), diacetyl (0.59%), and esters
(15.97%). Fermented nipa sap contained alcohols (91.16 – 98.29%), esters (1.18 – 8.14%), acetoin (0.02 – 0.7%), diacetyl (0.04 – 0.06%), and acetic acid (0.13 – 0.68%). Concentration of ethanol in fresh nipa sap increased from 0.11% (v/v) to 6.63% (v/v) during the fermentation, and slightly decreased to 5.73% (v/v) at day 63. No higher alcohols were detected in the fresh nipa sap. Concentration of 1-propanol and 2-methylpropanol were constant throughout the fermentation with average of 0.004 to 0.006% (v/v) and 0.0001 to 0.0009% (v/v), respectively. 3-methylbutanol increased during the fermentation process. The highest concentration (0.001% v/v) was recorded at day 35. This study has shown differences in VOCs types between fresh and fermented nipa sap.
A study was carried out to observe the fermentation process for noni (Morinda citrifolia L.) extract by Saccharomyces cerevisiae. The experiment was based on a central composite rotatable design (CCRD) employing 5 center points with augmented axial and factorial points resulting in 30 runs. The M. citrifolia extract was fermented with different combination of substrate concentration (40, 50, 60, 70 and 80%) (w/v), inoculum size (0, 1.5, 3, 4.5 and 6%) (v/v), temperature (30, 33.5, 37, 40.5 and 44oC) and fermentation time (0, 1.5, 3, 4.5 and 6 days). Five physico-chemical characteristics which include pH, titratable acidity, turbidity, total soluble solids and total polyphenol content were measured. Results showed that all the responses could be well represented using statistical models. For pH, only fermentation time was found to be not significant, while for titratable acidity and total polyphenol content, the effects of substrate concentration and fermentation time were significant. The effects of inoculum size and temperature level were found to be significant for turbidity. For total soluble solids, only the effect of substrate concentration and inoculum size were found to be significant.
Megabiodiversity of Malaysian’s flora and fauna which include microorganism could be conserved and served as alternative source indigenous yeast, the leavening agent of commercial bread making. This study was conducted in attempt to exploit the potential of Saccharomyces cerevisiae strains isolated from 30 different local fruits and plant parts as a leavening agent in bread making. The enrichment was carried out by fermenting the plant samples in medium containing Grape Must at 25°C for 10 days following by isolation of tentative yeasts at 30°C for 3 to 5 days. 20 out of 30 samples tested showed the presence of yeasts was then selected for identification of S. cerevisiae strains through biochemical and physiological tests. Of the 20 yeast strains examined, 13 strains were identified as S. cerevisiae and potentially used as leavening agent in bread making where 5 strains namely SN3, SMK9, SDB10, SRB11 and SS12 showed better fermentative performance compared to commercial strains. Thus, indicated that the local fruits and plant parts could be the potential source of indigenous S. cerevisiae strains for leavening agent in bread making.
Sustainable, environmental friendly, and safe disposal of sewage treatment plant (STP) sludge is a global expectation. Bioremediation performance was examined at different hydraulic retention times (HRT) in 3-10 days and organic loading rates (OLR) at 0.66-7.81 g chemical oxygen demand (COD) per liter per day, with mixed filamentous fungal (Aspergillus niger and Penicillium corylophilum) inoculation by liquid-state bioconversion (LSB) technique as a continuous process in large-scale bioreactor. Encouraging results were monitored in treated sludge by LSB continuous process. The highest removal of total suspended solid (TSS), turbidity, and COD were achieved at 98, 99, and 93%, respectively, at 10 days HRT compared to control. The minimum volatile suspended solid/suspended solid implies the quality of water, which was recorded 0.59 at 10 days and 0.72 at 3 days of HRT. In treated supernatant with 88% protein removal at 10 days of HRT indicates a higher magnitude of purification of treated sludge. The specific resistance to filtration (SRF) quantifies the performance of dewaterability; it was recorded minimum 0.049 × 10(12) m kg(-1) at 10 days of HRT, which was equivalent to 97% decrease of SRF. The lower OLR and higher HRT directly influenced the bioremediation and dewaterability of STP sludge in LSB process. The obtained findings imply encouraging message in continuing treatment of STP sludge, i.e., bioremediation of wastewater for environmental friendly disposal in near future.
The feasibility of using dried attached-growth biomass from the polyurethane (PU) foam cubes as a solid carbon source to enhance the denitrification process in the intermittently aerated moving bed sequencing batch reactor (IA-MBSBR) during the treatment of low COD/N containing wastewater was investigated. By packing the IA-MBSBR with 8% (v/v) of 8-mL PU foam cubes saturated with dried attached-growth biomass, total nitrogen removal efficiency of 80% could be achieved for 10 consecutive cycles of operation when the intermittent aeration strategy of consecutive 1 h of aeration followed by 2 h of non-aeration period during the REACT period of the IA-MBSBR was adopted. Negligible release of ammonium nitrogen (NH4(+)-N) and slow-release of COD from the dried biomass would ensure that the use of this solid carbon source would not further burden the treatment system. The slow-releasing COD was found to have no effect in promoting the assimilation process and would also allow the carbon source to be used for many cycles of operation. The 'carbon-spent' PU foam cubes could be reused by merely drying at 60 °C at the end of the operational mode. Thus, the dried attached-growth biomass formed on the PU foam cubes could be exploited as an alternative solid carbon source for the enhancement of denitrification process in the IA-MBSBR.
Recent years, great interest has been devoted to the conversion of biomass-derived carbohydrate into sugars, such as glucose, mannose and fructose. These are important versatile intermediate products that are easily processed into high value-added biofuels. In this work, microwave-assisted dilute sulfuric acid hydrolysis of deproteinated palm kernel cake (DPKC) was systematically studied using Response Surface Methodology. The highest mannose yield (92.11%) was achieved at the optimized condition of 148°C, 0.75N H2SO4, 10min 31s and substrate to solvent (SS) ratio (w/v) of 1:49.69. Besides that, total fermentable sugars yield (77.11%), was obtained at 170°C, 0.181N H2SO4, 6min 6s and SS ratio (w/v) of 1:40. Ridge analysis was employed to further verify the optimum conditions. Thus, this work provides fundamental data of the practical use of DPKC as low cost, high yield and environmental-friendly material for the production of mannose and other sugars.
Production of extracellular laccase by the white-rot fungus Pycnoporus sanguineus was examined in batch submerged cultures in shake flasks, baffled shake flasks and a stirred tank bioreactor. The biomass growth in the various culture systems closely followed a logistic growth model. The production of laccase followed a Luedeking-Piret model. A modified Luedeking-Piret model incorporating logistic growth effectively described the consumption of glucose. Biomass productivity, enzyme productivity and substrate consumption were enhanced in baffled shake flasks relative to the cases for the conventional shake flasks. This was associated with improved oxygen transfer in the presence of the baffles. The best results were obtained in the stirred tank bioreactor. At 28 °C, pH 4.5, an agitation speed of 600 rpm and a dissolved oxygen concentration of ~25 % of air saturation, the laccase productivity in the bioreactor exceeded 19 U L(-1 )days(-1), or 1.5-fold better than the best case for the baffled shake flask. The final concentration of the enzyme was about 325 U L(-1).