Assessment of microbial activity by CO2 production during heating oil storage

Microbial activity is the driving force of the carbon cycle, including the digestion of biomass in the soil, oceans, and oil deposits. This natural diversity of microbial carbon sources poses challenges for humans. Contamination monitoring can be difficult in oil tanks and similar settings. To assess microbial activity in such industrial settings, off‐gas analysis can be employed by considering growth and non‐growth‐associated metabolic activity. In this work, we describe the monitoring of CO₂ as a method for measuring microbial activity. We revealed that the CO₂ signal corresponds to classical growth curves, exemplified by Pseudomonas fluorescens, Yarrowia lipolytica, and Penicillium chrysogenum. Deviations of the CO₂ signal from the growth curves occurred when the yield of biomass on the substrate changed (i.e., the non‐growth‐associated metabolic activities). We monitored CO₂ to track the onset of microbial contamination in an oil tank. This experimental setup was applied to determine the susceptibility of heating oil and biodiesel to microbial contamination long before the formation of problematic biofilms. In summary, the measurement of CO₂ production by bacteria, yeasts, and molds allowed the permanent monitoring of microbial activity under oil storage conditions without invasive sampling.     

Start-up of an UASB-septic tank for community on-site treatment of strong domestic sewage

Two community on-site UASB-septic tanks were operated in parallel over a six months period under two different hydraulic retention times (HRT) of 2 days for R1 and 4 days for R2 at mean sewage temperature of 24 degrees C. The sewage was characterised by a high COD(tot) concentration of 1189 mg/L, with a large fraction of COD(sus), viz. 54%. The achieved removal efficiencies in R1 and R2 for COD(tot), COD(sus), BOD5 and TSS were "56%, 87%, 59% and 81%" and "58%, 90%, 60% and 82%" for both systems, respectively. R2 achieved a marginal but significant (p<0.05) better removal efficiencies of those parameters as compared to R1. The COD(col) and COD(dis) removals in R1 and R2 were respectively 31% and 20%, and 34% and 22%. The sludge accumulation was very low suggesting that the desludging frequency will be of several years. Accordingly, the reactor can be adequately designed at 2 days HRT.     

Microbial oil production from rice straw hydrolysate by Trichosporon fermentans

Microbial oil production from sulphuric acid treated rice straw hydrolysate (SARSH) by Trichosporon fermentans was performed for the first time. Fermentation of SARSH without detoxification gave a poor lipid yield of 1.7 g/l, which was much lower than the result with glucose or xylose as the single carbon source (13.6 g/l or 9.9 g/l). The detoxification pretreatment, including overliming, concentration, and adsorption by Amberlite XAD-4 improved the fermentability of SARSH significantly by removing the inhibitors in SARSH. A total biomass of 28.6 g/l with a lipid content of 40.1% (corresponding to a lipid yield of 11.5 g/l) could be achieved after cultivation of T. fermentans on the detoxified SARSH for 8 days. Moreover, besides SARSH, T. fermentans could also utilize mannose, galactose, or cellobiose, in hydrolysates of other natural lignocellulosic materials as the single carbon source to grow and accumulate lipid with a high yield (at least 10.4 g/l). Hence, it is a promising strain for microbial oil production and thus biodiesel preparation from agro-industrial residues, especially lignocellulosic materials.     

Evaluation of single cell oil from Aureobasidium pullulans var. melanogenum P10 isolated from mangrove ecosystems for biodiesel production

In this study, the yeast strain P10 which was identified to be a member of Aureobasidium pullulans var. melanogenum isolated from the mangrove ecosystems was found to be able to accumulate high content of oil in its cells. After optimization of the medium for lipid production and cell growth by the yeast strain P10, it was found that 8.0 g of glucose per 100 ml, 0.02 g of yeast extract per 100 ml, 0.02 g of ammonium sulfate per 100 ml, pH 6.0 in the medium were the most suitable for lipid production. During 10-l fermentation, a titer was 66.3 g oil per 100 g of cell dry weight, cell mass was 1.3 g per 100 ml, a yield was 0.11 g of oil per g of consumed sugar and a productivity was 0.0009 g of oil per g of consumed sugar per h within 120 h. At the same time, only 0.07 g of reducing sugar per 100 ml was left in the fermented medium. The compositions of the fatty acids produced were C_16:0 (26.7%), C_16:1(1.7%), C_18:0 (6.1%), C_18:1 (44.5%), and C_18:2 (21.0%). The biodiesel produced from the extracted lipid could be burnt well.     

Silica-bonded N-propyl sulfamic acid used as a heterogeneous catalyst for transesterification of soybean oil with methanol

The transesterification of soybean oil with methanol was carried out, to produce biodiesel, over silica-bonded N-propyl sulfamic acid in a heterogeneous manner. Results showed that a maximum conversion of 90.5% was achieved using a 1:20 M ratio of soybean oil to methanol and a catalyst amount of 7.5 wt.% at 423 K for 60 h. It was found that the free fatty acid (FFA) and water present in the feedstock had no significant influence on the catalytic activity to the transesterification reaction. Besides, the catalyst also showed activities towards the esterification reaction of FFAs, in terms of the FFA conversion of 95.6% at 423 K for 30 h. Furthermore, the catalyst could be recovered with a better reusability.     

Novozym 435 for production of biodiesel from unrefined palm oil: Comparison of methanolysis methods

Biodiesel (BD) is commonly produced from refined vegetable oils by alkali-catalyzed methanolysis. Unrefined vegetable oils are economically attractive but not suitable for alkali catalysis because of their high content of free fatty acids (FFAs). Novozym 435 (immobilized Candida antarctica lipase B), which accepts both FFA and oil as substrates, was, therefore, employed to convert unrefined palm oil to BD. Three different methanolysis methods, namely, t-butanol mediated system (method-1), LiCl solution based controlled release system for methanol (method-2) and solvent-free system with three successive additions of methanol (method-3), were compared. The optimal methanol to oil molar ratios in the method-1, -2 and -3 are 6:1, 3:1 and 3:1, respectively. BD yield at an optimal methanol concentration reaches 91–92% after 10, 20 and 24 h in the method-1, -2 and -3, respectively. BD yield remains the same over five repeated cycles in the method-1, while it drops to 68 and 71% by the fifth cycle in the method-2 and -3, respectively. The results show that the method-1 is the most effective for production of BD from a low cost feedstock like unrefined palm oil.     

Ethanesulfonic acid-based esterification of industrial acidic crude palm oil for biodiesel production

An industrial grade acidic crude palm oil (ACPO) pre-treatment process was carried out using ethanesulfonic acid (ESA) as a catalyst in the esterification reaction. ESA was used in different dosages to reduce free fatty acid (FFA) to a minimum level for the second stage of biodiesel production via alkaline transesterification reaction. Different process operating conditions were optimized such as ESA dosage (0.25-3.5% wt/wt), methanol to ACPO molar ratio (1:1-20:1), reaction temperature (40-70 °C), and reaction time (3-150 min). This study revealed the potential use of abundant quantities of ACPO from oil palm mills for biodiesel production. The lab scale results showed the effectiveness of the pre-treatment process using ESA catalyst. Three consecutive catalyst recycling runs were achieved without significant degradation in its performance. Second and third reuse runs needed more reaction time to achieve the target level of FFA content. Esterification and transesterification using ESA and KOH respectively is proposed for biodiesel industrial scale production. The produced biodiesel meets the international standards specifications for biodiesel fuel (EN 14214 and ASTM D6751).     

Homogeneous,heterogeneous and enzymatic catalysis for transesterification of high free fatty acid oil (waste cooking oil) to biodiesel: A review

In the last few years, biodiesel has emerged as one of the most potential renewable energy to replace current petrol-derived diesel. It is a renewable, biodegradable and non-toxic fuel which can be easily produced through transesterification reaction. However, current commercial usage of refined vegetable oils for biodiesel production is impractical and uneconomical due to high feedstock cost and priority as food resources. Low-grade oil, typically waste cooking oil can be a better alternative; however, the high free fatty acids (FFA) content in waste cooking oil has become the main drawback for this potential feedstock. Therefore, this review paper is aimed to give an overview on the current status of biodiesel production and the potential of waste cooking oil as an alternative feedstock. Advantages and limitations of using homogeneous, heterogeneous and enzymatic transesterification on oil with high FFA (mostly waste cooking oil) are discussed in detail. It was found that using heterogeneous acid catalyst and enzyme are the best option to produce biodiesel from oil with high FFA as compared to the current commercial homogeneous base-catalyzed process. However, these heterogeneous acid and enzyme catalyze system still suffers from serious mass transfer limitation problems and therefore are not favorable for industrial application. Nevertheless, towards the end of this review paper, a few latest technological developments that have the potential to overcome the mass transfer limitation problem such as oscillatory flow reactor (OFR), ultrasonication, microwave reactor and co-solvent are reviewed. With proper research focus and development, waste cooking oil can indeed become the next ideal feedstock for biodiesel.     

Biochemical and kinetic evaluation of lipase and biosurfactant assisted ex novo synthesis of microbial oil for biodiesel production by Yarrowia lipolytica utilizing chicken tallow

The present study explores the production of biodiesel, a sustainable replacement for depleting fossil fuel by utilizing microbial oil, which was procured from Yarrowia lipolytica employing chicken tallow as the carbon substrate. Chicken tallow, yeast extract, and MgSO₄·7H₂O were screened for biomass production through Plackett–Burman design. Further, Box–Behnken design analysis was performed, and the optimal concentration of the medium variables was found to be 20 g/L of chicken tallow, 7.0 g/L of yeast extract, and 0.45 g/L of MgSO₄·7H₂O.The various parameters viz., pH (6), temperature (30 °C), RPM (150), inoculum volume (5%, v/v), and C/N ratio (100) were optimized for maximal biomass and lipid yield, and lipid content. Nile red-stained cells were observed for intracellular lipid bodies using fluorescence microscopy, and its fluorescence intensity was measured bythe flow cytometer. The dimorphic transition and substrate assimilation of Y. lipolytica were analyzed using scanning electron microscopy (SEM), and Fourier transform infrared spectroscopy (FT-IR). Batch kinetic studies revealed the concomitant synthesis of microbial lipid (4.16 g/L), lipase (43 U/mL), and biosurfactant (1.41 g/L). The GC-MS analysis of microbial oil presented the fatty acid profile as oleic acid (49.15%), palmitic acid (29.83%), stearic acid (11.43%), linoleic acid (3.83%), palmitoleic acid (3.77%), and myristic acid (1.32%).     

Preparation of cross-linked lipase-coated micro-crystals for biodiesel production from waste cooking oil

A dual modification procedure composed of cross-linking and protein coating with K₂SO₄ was employed to modify Geotrichum sp. lipase for catalyzing biodiesel production from waste cooking oil. Compared to single modification of protein coating with K₂SO₄, the dual modification of cross-linking and lipase coating improved catalytic properties in terms of thermostable stability, organic solvent tolerance, pH stability and operational stability in biodiesel production process, although biodiesel yield and initial reaction rate for CLPCMCs were not improved. After five successive batch reactions, CLPCMCs could still maintain 80% of relative biodiesel yield. CLPCMCs retained 64% of relative biodiesel yield after incubation in a pH range of 4-6 for 4 h, and 85% of relative biodiesel yield after incubation in a range of 45-50 °C for 4 h. CLPCMCs still maintained 83% of relative biodiesel yield after both treated in polar organic solvent and non-polar organic solvent for 4 h.     

Microbial consortia for the aerobic degradation of aromatic compounds in olive oil mill effluent

Aerobic consortia that grow on olive oil mill effluent (OOME) were obtained by enrichment. Several cultures were capable of metabolizing monoaromatic compounds, supplied as the sole carbon source at 2 g L^–1. Some consortia degraded mixtures of seven aromatics (4 g L^–1) after 1 week of incubation at 32°C. The consortia were also active against monoaromatics of the undiluted OOME. This reduced the inhibitory effect of phenolic compounds prior to the anaerobic digestion of OOME at batch scale. No inhibition of the anaerobic microbial populations was noticed with treated OOME. From the most active consortium, nine different bacterial strains were isolated and shown to grow on simple aromatic compounds. Removal of 50% of the initial chemical oxygen demand and degradation of almost all of the simple aromatics in undiluted OOME was obtained with reconstituted bacterial mixtures. A slight reduction in colouration was due to adsorption of coloured compounds to bacterial cells. Presumably, the consortia could not reduce and degrade the coloured compounds in OOME.     

Projected climate change effects on Alberta's boreal forests imply future challenges for oil sands reclamation

Climate change will drive significant changes in vegetation cover and also impact efforts to restore ecosystems that have been disturbed by human activities. Bitumen mining in the Alberta oil sands region of western Canada requires reclamation to “equivalent land capability,” implying establishment of vegetation similar to undisturbed boreal ecosystems. However, there is consensus that this region will be exposed to relatively severe climate warming, causing increased occurrence of drought and wildfire, which threaten the persistence of both natural and reclaimed ecosystems. We used a landscape model, LANDIS‐II, to simulate plant responses to climate change and disturbances, forecasting changes to boreal forests within the oil sands region. Under the most severe climate forcing scenarios (representative concentration pathway [RCP] 8.5) the model projected substantial decreases in forest biomass, with the future forest being dominated by drought‐ and fire‐tolerant species characteristic of parkland or prairie ecosystems. In contrast, less extreme climate forcing scenarios (RCPs 2.6 and 4.5) had relatively minor effects on forest composition and biomass with boreal conifers continuing to dominate the landscape. If the climate continues to change along a trajectory similar to those simulated by climate models for the RCP 8.5 forcing scenario, current reclamation goals to reestablish spruce‐dominated boreal forest will likely be difficult to achieve. Results from scenario modeling studies such as ours, and continued monitoring of change in the boreal forest, will help inform reclamation practices, which could include establishment of species better adapted to warmer and drier conditions.     

Screening Botryosphaeria species for lipases: Production of lipase by Botryosphaeria ribis EC-01 grown on soybean oil and other carbon sources

Nine isolates of Botryosphaeria spp. were screened for lipases when cultivated on eight different plant seed oils and glycerol, and all produced lipases. Botryosphaeria ribis EC-01 produced highest lipase titres on soybean oil and glycerol, while eight isolates of Botryosphaeria rhodina produced significantly lower enzyme titres. B. ribis EC-01 produced lipase when grown on different fatty acids, surfactants, carbohydrates and triacylglycerols, with highest enzyme titres produced on Triton X-100-emulsified stearic (316.7 U/mL), palmitic (283.5 U/mL) and oleic (247.4 U/mg) acids, and soybean oil (105.6 U/mL), as well as castor oil (191.2 U/mg); an enhancement of 9-fold over soybean oil-grown cultures. Glycerol was also a good substrate for lipase production. The crude lipase extract was optimally active at pH 8.0 and 55 °C, stable between 30 and 55 °C and pH 1–10, and tolerant to 50% (v/v) glycerol, methanol and ethanol. The crude lipase showed affinity for substrates of short, average and long-chain fatty acids (different esters of p-nitrophenol and triacylglycerols). Zymograms developed with 4-methylumbelliferyl-butyrate showed two bands of lipolytic activity at 45 and 15 kDa. This is the first report on the production of lipases by B. ribis grown on these different carbon sources.