Energetic assessment of fixation of CO<Subscript>2</Subscript> and subsequent biofuel production using <Emphasis Type="Italic">B. cereus</Emphasis> SM1 isolated from sewage treatment plant

The ongoing work on global warming resulting from green house gases (GHGs) has led to explore the possibility of bacterial strains which can fix carbon dioxide (CO₂) and can generate value-added products. The present work is an effort in this direction and has carried out an exhaustive batch experiments for the fixation of CO₂ using B. Cereus SM1 isolated from sewage treatment plant (STP). The work has incorporated 5-day batch run for gaseous phase inlet CO₂ concentration of 13 ± 1 % (%v/v). 84.6 (±5.76) % of CO₂ removal was obtained in the gaseous phase at mentioned CO₂ concentration (%v/v). Energetic requirement for CO₂ fixation was assessed by varying Fe[II] ion concentration (0–200 ppm) on the per-day basis. The cell lysate obtained from CO₂ fixation studies (Fe[II] ion = 100 ppm) was analyzed using Fourier transformation infrared spectroscopy (FTIR) and gas chromatography-mass spectroscopy (GC–MS). This analysis confirmed the presence of fatty acids and hydrocarbon as valuable products. The hydrocarbons were found in the range of C₁₁–C₂₂ which is equivalent to light oil. The obtained fatty acids were found in the range of C₁₁–C₁₉. The possibility of fatty acid conversion to biodiesel was explored by carrying out the transesterification reaction. The yield of biodiesel was obtained as 86.5 (±0.048) % under the transesterification reaction conditions. Results of this research work can provide the valuable information in the implementation of biomitigation of CO₂ at real scenario.     

Improvement of cell-bound lipase from Rhodotorula mucilaginosa P11I89 for use as a methanol-tolerant, whole-cell biocatalyst for production of palm-oil biodiesel

Rhodotorula mucilaginosa P11I89, isolated from oil-contaminated soil, was effectively used as the methanol-tolerant, whole-cell lipase for the synthesis of fatty acid methyl ester (FAME) via transesterification reaction in the presence of palm oil and methanol substrates at a 1:6 mole ratio. A combination of Taguchi experimental design and response surface methodology (RSM) were applied to systemically enhance transesterification activity of the whole-cell lipase or cell-bound lipase (CBL) from R. mucilaginosa P11I89 in a solvent-free system. The significant impacts of four factors including carbon sources, nitrogen sources, surfactants and pH on hydrolysis activity of extracellular and cell-bound lipases, and on the transesterification activity of CBL were evaluated using Taguchi design. Gum Arabic was the most significant component for high transesterification activity, whereas soybean oil was the most influential factor for the hydrolysis activity. Maximal CBL production of 272.72 U/L was obtained in the cultivation medium containing 2.1 % palm oil, 0.2 % NH₄NO₃ , and 0.45 % Gum Arabic, with initial pH 5.0 under shaking speed of 200 rpm at a temperature of 30?±?2 °C after 60 h incubation using Central Composite Design (CCD). Yeast cells grown under such conditions increased FAME yield from 84.0 to 92.98 % when the transesterification reaction was carried out, in comparison to those cultivated in the initial medium.     

A packed bed membrane reactor for production of biodiesel using activated carbon supported catalyst

In this study, a novel continuous reactor has been developed to produce high quality methyl esters (biodiesel) from palm oil. A microporous TiO₂/Al₂O₃ membrane was packed with potassium hydroxide catalyst supported on palm shell activated carbon. The central composite design (CCD) of response surface methodology (RSM) was employed to investigate the effects of reaction temperature, catalyst amount and cross flow circulation velocity on the production of biodiesel in the packed bed membrane reactor. The highest conversion of palm oil to biodiesel in the reactor was obtained at 70 °C employing 157.04 g catalyst per unit volume of the reactor and 0.21 cm/s cross flow circulation velocity. The physical and chemical properties of the produced biodiesel were determined and compared with the standard specifications. High quality palm oil biodiesel was produced by combination of heterogeneous alkali transesterification and separation processes in the packed bed membrane reactor.     

Potential for optimized production and use of rapeseed biodiesel. Based on a comprehensive real-time LCA case study in Denmark with multiple pathways

Purpose

Several factors contribute to the current increased focus on alternative fuels such as biodiesel, including an increasing awareness of the environmental impact of petrochemical (PC) oil products such as PC diesel, the continuously increasing price of PC oil, and the depletion of PC oil. For these reasons, the European Union has enacted a directive requiring each member state to ensure that the share of energy from renewable sources in transport be at least 10 % of the final consumption of energy by 2020 (The European Parliament and the Council 2009). This LCA study assesses the specific environmental impacts from the production and use of biodiesel as it is today (real-time), based on rapeseed oil and different types of alcohols, and using technologies that are currently available or will be available shortly. Different options are evaluated for the environmental improvement of production methods. The modeling of the LCA is based on a specific Danish biodiesel production facility.

Methods

The functional unit is “1,000 km transportation for a standard passenger car.” All relevant process stages are included, such as rapeseed production including carbon sequestration and N₂O balances, and transportation of products used in the life cycle of biodiesel. System expansion has been used to handle allocation issues.

Results and discussion

The climate change potential from the production and use of biodiesel today is 57 kg CO₂-eq/1,000 km, while PC diesel is 214 kg CO₂-eq/1,000 km. Options for improvement include the increased use of residual straw from rapeseed fields for combustion in a power plant where carbon sequestration is considered, and a change in transesterification from a conventional process to an enzymatic process when using bioethanol instead of PC methanol. This research also evaluates results for land use, respiratory inorganics potential, human toxicity (carc) potential, ecotoxicity (freshwater) potential, and aquatic eutrophication (N) potential. Different sources for uncertainty are evaluated, and the largest drivers for uncertainty are the assumptions embedded into the substitution effects. The results presented should not be interpreted as a blueprint for the increased production of biodiesel but rather as a benchmarking point for the present, actual impact in a well-to-wheels perspective of biodiesel, with options for improving production and use.

Conclusions

Based on this analysis, we recommend investigating additional options and incentives regarding the increased use of rape straw, particularly considering the carbon sequestration issues (from the perspective of potential climate change) of using bioalcohol instead of PC alcohol for the transesterification process.     

Batch and fed-batch growth of Pichia pastoris under increased air pressure

Pichia pastoris CBS 2612 behavior under air pressures of 1, 3 and 5 bar in culture media of glycerol (pure and crude) and methanol was studied. Generally, the increase in oxygen transfer rate due to the increase of total pressure improved cellular growth for all carbon sources and for batch and fed-batch processes with different feeding rate strategies. In batch cultures, 1.4-, 1.2-, and 1.5-fold improvement in biomass production was obtained with the increase of air pressure up to 5 bar, using methanol, pure glycerol, and crude glycerol, respectively. The increase of air pressure to 5 bar using exponential feeding rate led to 1.4-fold improvement in biomass yield per glycerol mass consumed, for crude and pure glycerol. The current low cost of crude glycerol from the biodiesel production together with the present results shows the possibility of improving cell mass production of P. pastoris using increased air pressure.     

Testing Refrigeration Trucks for the Emergency Evacuation of Companion Animals

The purpose of this study was to quantify the changes in oxygen (O₂) and carbon dioxide (CO₂) in sealed refrigerator trucks scheduled to be used for transporting companion animals (dogs and cats) during an emergency evacuation. A total of 122 nonhuman animals (total weight = 1,248 kg) housed in individual crates were loaded into a 16-m refrigeration truck. Once they were loaded, the doors were closed and the percentages of O₂ and CO₂ were measured every 5 min by O₂ and CO₂ analyzers, and they were used to quantify the changes in gas pressure in the sealed truck. CO₂ had a much higher-than-predicted increase, and O₂ had a higher-than-predicted decrease. These 2 pressures in combination with the functionality of the respiratory system will limit the animal's ability to load O₂, and over time, they will initiate asphyxia or suffocation. Over time, the partial pressure of oxygen (P_O2) in the sealed truck will decrease, causing hypoxia, and the partial pressure of carbon dioxide (P_CO2) will increase, causing hypercapnia.     

Efficient solvothermal wet in situ transesterification of Nannochloropsis gaditana for biodiesel production

In situ transesterification of wet microalgae is a promising, simplified alternative biodiesel production process that replaces multiple operations of cell drying, extraction, and transesterification reaction. This study addresses enhanced biodiesel production from Nannochloropsis gaditana at elevated temperatures. Compared with the previously reported in situ transesterification process of conducting the reaction at a temperature ranging from 95 to 125 °C, the present work employs higher temperatures of at least 150 °C. This relatively harsh condition allows much less acid catalyst with or without co-solvent to be used during this single extraction-conversion process. Without any co-solvent, 0.58% (v/v) of H₂SO₄ in the reaction medium can achieve 90 wt% of the total lipid conversion to biodiesel at 170 °C when the moisture content of wet algal paste is 80 wt%. Here, the effects of temperature, acid catalyst, and co-solvent on the FAEE yield and specification were scrutinized, and the reaction kinetic was investigated to understand the solvothermal in situ transesterification reaction at the high temperature. Having a biphasic system (water/chloroform) during the reaction also helped to meet biodiesel quality standard EN 14214, as Na⁺, K⁺, Ca²⁺, Mg²⁺ cations and phosphorus were detected only below 5 ppm. With highlights on the economic feasibility, wet in situ transesterification at the high temperature can contribute to sustainable production of biodiesel from microalgae by reducing the chemical input and relieve the burden of extensive post purification process, therefore a step towards green process.

     

The effects of supercritical carbon dioxide extraction and cold-pressed hexane extraction on the chemical composition and feeding value of rapeseed meal for broiler chickens

ABSTRACT

The chemical characteristics of rapeseed meal (RSM) produced from two cultivars of UK-grown rapeseed, by both supercritical carbon dioxide extraction (ScCO₂) and cold-pressed hexane extraction (CpHe) were examined. Their nutritional value, with and without protease, was then assessed in a broiler digestibility trial. Basal feed was substituted with one of four RSM batches (200 g/kg) following adjustments for dry matter (DM) and ether extract (EE) content. Half of each diet was supplemented with a mono-component protease derived from Bacillus subtilis (Axtra®PRO, Danisco Animal Nutrition, Malborough, UK) giving a total of eight test diets. Two control diets, with and without protease were also fed. At 13 d age male Ross 308 broilers were randomly allocated to seven replicate pens (five birds per pen) and assigned to one of 10 diets. Total excreta were collected from 17 to 21 d age and feed intake was recorded. Pre-caecal protein digestibility (pcPd) was determined using TiO₂ as an indigestible marker. Colourimetrically CpHe RSM was substantially darker than ScCO₂ counterparts. The influence of oil recovery method (ORM) was also evident in DM, EE, ash free neutral detergent fibre (aNDFom), neutral detergent insoluble crude protein (NDICP) and glucosinolate content (GLS). The content of DM, EE and GLS was higher in ScCO₂ RSM whereas aNDFom and NDICP levels were greater in CpHe RSM. Protein solubility in KOH was greater in ScCO₂ RSM whilst levels of NDICP were lower. Collectively these results suggest that less heat damage was incurred to the RSM during ScCO₂ extraction. There was no significant main effect of cultivar nor were any significant interactions observed between treatment factors. Rapeseed meal ScCO₂ produced greater metabolisable energy, pcPd, nitrogen retention and energy metabolisability (p < 0.05). Protease supplementation increased pcPd (p < 0.05) irrespective of ORM and cultivar. The key implications of these findings are that by adopting oil recovery methods that minimise the exposure of RSM to thermal treatments and by adding a compatible protease there is scope to increase the nutritional value of RSM for broilers and increase its utilisation in modern poultry production.     

Continuous enzymatic biodiesel production from coconut oil in two-stage packed-bed reactor incorporating an extracting column to remove glycerol formed as by-product

The transesterification of coconut oil with ethanol catalyzed by Burkholderia cepacia lipase immobilized on polysiloxane–polyvinyl alcohol was performed in a continuous flow. The experimental design consisted of a two-stage packed-bed reactor incorporating a column with cationic resin (Lewatit GF 202) to remove the glycerol formed as by-product and the reactor performance was quantified for three different flow rates corresponding to space-times from 10 to 14 h. The influence of space-time on the ethyl ester (FAEE) concentrations, yields and productivities was determined. The reactor operation was demonstrated for space-time of 14 h attaining FAEE concentrations of 58.5 ± 0.87 wt%, FAEE yields of 97.3 ± 1.9 % and productivities of 41.6  ± 1.0 mg_ester g _medium ^?1  h^?1. Biodiesel purified samples showed average kinematic viscosity values of 5.5 ± 0.3 mm² s^?1 that meet the criteria established by the American National Standard ASTM (D6751). The immobilized lipase was found to be stable regarding its morphological and catalytic characteristics, showing half-life time (t _1/2) around 1540 h. The continuous packed-bed reactor connected in series with simultaneous glycerol removal has a great potential to attain high level of transesterification yields, raising biodiesel productivity.     

Carbon dioxide utilisation of Dunaliella tertiolecta for carbon bio-mitigation in a semicontinuous photobioreactor

Bio-fixation of carbon dioxide (CO₂) by microalgae has been recognised as an attractive approach to offset anthropogenic emissions. Biological carbon mitigation is the process whereby autotrophic organisms, such as microalgae, convert CO₂ into organic carbon and O₂ through photosynthesis; this process through respiration produces biomass. In this study Dunaliella tertiolecta was cultivated in a semicontinuous culture to investigate the carbon mitigation rate of the system. The algae were produced in 1.2-L Roux bottles with a working volume of 1 L while semicontinuous production commenced on day 4 of cultivation when the carbon mitigation rate was found to be at a maximum for D. tertiolecta. The reduction in CO₂ between input and output gases was monitored to predict carbon fixation rates while biomass production and microalgal carbon content are used to calculate the actual carbon mitigation potential of D. tertiolecta. A renewal rate of 45 % of flask volume was utilised to maintain the culture in exponential growth with an average daily productivity of 0.07 g L^?1 day^?1. The results showed that 0.74 g L^?1 of biomass could be achieved after 7 days of semicontinuous production while a total carbon mitigation of 0.37 g L^?1 was achieved. This represented an increase of 0.18 g L^?1 in carbon mitigation rate compared to batch production of D. tertiolecta over the same cultivation period.     

Biodiesel production from rice bran oil and supercritical methanol

In this study, production of biodiesel from low cost raw materials, such as rice bran and dewaxed-degummed rice bran oil (DDRBO), under supercritical condition was carried out. Carbon dioxide (CO₂) was employed as co-solvent to decrease the supercritical temperature and pressure of methanol. The effects of different raw materials on the yield of biodiesel production were investigated. In situ transesterification of rice bran with supercritical methanol at 30 MPa and 300 °C for 5 min was not a promising way to produce biodiesel because the purity and yield of fatty acid methyl esters (FAMEs) obtained were 52.52% and 51.28%, respectively. When DDRBO was reacted, the purity and yield were 89.25% and 94.84%, respectively. Trans-FAMEs, which constituted about 16% of biodiesel, were found. They were identified as methyl elaidate [trans-9], methyl linoleaidate [trans-9, trans-12], methyl linoleaidate [cis-9, trans-12], and methyl linoleaidate [trans-9, cis-12]. Hydrocarbons, which constituted about 3% of the reaction product, were also detected.     

Sonication-assisted production of biodiesel using soybean oil and supercritical methanol

High temperature and pressure are generally required to produce biodiesel using supercritical methanol. We reduced the harsh reaction conditions by means of sonicating the reaction mixture prior to transesterification using supercritical methanol. Soybean oil was selected as the raw material for transesterification. As soybean oil contains more unsaturated fatty acid triglycerides, the biodiesel degraded more at high temperature. The reactants were sonicated for 60 min at 35 °C prior to transesterification to avoid degradation of the product and to enhance biodiesel yield at temperatures <300 °C. The process parameters were optimized using central composite design. The variables selected for optimization were temperature, time, and the oil to methanol molar ratio. The temperature and oil to methanol molar ratios were varied from 250 to 280 °C and 1:40–1:50, respectively. The reaction time was tested between 4 and 12 min. The biodiesel was analyzed for any possible degradation by gas chromatography–mass spectroscopy and for the wt% of fatty acid methyl esters (FAME) obtained. The maximum FAME yield (84.2 wt%) was obtained at a temperature of 265.7 °C, an oil to alcohol molar ratio of 1:44.7, and a time of 8.8 min. The optimum yield was obtained at a pressure of 1,500 psi. The pressure and optimum temperature used to obtain the maximum yield were the lowest reported so far without the use of a co-solvent. Thus, the severity of the supercritical reactions was reduced by adding sonication prior to the reaction.     

INVESTIGATION OF THE GIBBERELLIC ACID OPTIMIZATION WITH A STATISTICAL TOOL FROM Penicillium variable IN BATCH REACTOR

The culture conditions for gibberellic acid (GA₃) production by the fungus Penicillium variable (P. variable) was optimized using a statistical tool, response surface methodology (RSM). Interactions of culture conditions and optimization of the system were studied using Box–Behnken design (BBD) with three levels of three variables in a batch flask reactor. Experimentation showed that the model developed based on RSM and BBD had predicted GA₃ production with R ²  = 0.987. The predicted GA₃ production was optimum (31.57 mg GA₃/kg substrate) when the culture conditions were at 7 days of incubation period, 21% v/w of inoculum size, and 2% v/w of olive oil concentration as a natural precursor. The results indicated that RSM and BBD methods were effective for optimizing the culture conditions of GA₃ production by P. variable mycelia.     

Continuous Bioconversion of the Lipids of Corn,Wheat, and Triticale Distiller’s Dried Grains with Solubles to Biodiesel in Supercritical Carbon Dioxide and Characterization of the Products

Lipids of corn, wheat, and triticale distiller’s dried grains with solubles (DDGS) were extracted with supercritical carbon dioxide (SC-CO₂) and converted to biodiesel (fatty acid methyl esters; FAME) in a continuous SC-CO₂ bioreactor using immobilized lipase. Different properties of the products obtained were characterized. FAME content of the products ranged between 87.1 % (corn DDGS lipid biodiesel; CB) and 92.0 % (triticale DDGS lipid biodiesel; TB). CB had a higher viscosity (6.24 mm²/s) than those of wheat DDGS lipid biodiesel (WB) (5.97 mm²/s) and TB (5.67 mm²/s). Cloud point of CB, WB, and TB were ?8.48, ?6.52, and ?5.04 °C, respectively. Oxidative stability of WB and TB was higher than that of CB. Heat of combustion of products was 39.4–39.6 kJ/g. Continuous bioconversion of DDGS lipids to biodiesel in SC-CO₂ is a simple, efficient, and “green” alternative to the conventional process and targets new opportunities for the biodiesel industry.     

Kinetics of enzymatic transesterification and thermal deactivation using immobilized Burkholderia lipase as catalyst

The most effective way of enzymatic synthesis of biodiesel is through lipase-catalyzed transesterification, while its performance and economic feasibility should still be improved. In this study, lipase produced by an isolated Burkholderia sp. was immobilized on microsize Celite materials functionally modified with long alkyl groups. The specific activity of the immobilized lipase was 1,154 U/g. The methanolysis of olive oil catalyzed by the immobilized lipase obeyed Ping Pong Bi Bi model with an estimated V _max, K _m,TG, K _m,M and K _i,M value of 0.61 mol/(L min), 7.93 mol/L, 1.01 mol/L, and 0.24 mol/L, respectively. The activation energy of the enzymatic reaction is estimated as 15.51 kJ/mol. The immobilized lipase exhibits high thermal stability with thermal deactivation energy of 83 kJ/mol and a long half-life. The enthalpy, Gibb’s free energy, and entropy of the immobilized lipase were in the range of 80.02–80.35 kJ/mol, 88.35–90.13 kJ/mol, and ?28.22 to ?25.11 J/(mol K), respectively.     

Rapid transesterification of micro-amount of lipids from microalgae via a micro-mixer reactor

Background

Fatty acid composition is an important physiological parameter of microalgae, which is taken as the third generation alternative resource of biodiesel. To boost microalgal research and applications, a convenient, rapid, and acid-catalyzed transesterification procedure that satisfies the demand for the analysis of the fatty acid composition of lipids with micro-scale samples in the high-throughput screening of microalgal strains is needed, along with the evaluation of the physiological status of microalgae in response to nutrient stress.

Results

The reaction conditions of transesterification via a micro-mixer reactor were optimized as follows: 90 °C reaction temperature, 20 min reaction time, 6:1 volume ratio of H₂SO₄-methanol to lipid-in-hexane, and a Y-type micro-mixer with a 20-m-long extended loop that has a 0.3 mm diameter. The minimum amount of sample was decreased to 30 µg lipids. The new approach was successfully applied to the fatty acid composition analysis of soybean oil and microalgal lipids. Definitely, it could be applied to acyl related oils from different sources.

Conclusion

Here, we have developed a simple and rapid method for the analysis of the fatty acid composition of lipids. The new method requires less than 20 min for transesterification and a minimum of only 30 µg lipid sample. Furthermore, a high-throughput process can be easily realized by numbering up the micro-mixer reactors. The micro-mixer reactor has great potential for applications not only in large-scale biodiesel production but also for the micro-scale analysis of microalgae fatty acid compositions.

     

Synthesis of DHA/EPA-rich phosphatidylcholine by immobilized phospholipase A<Subscript>1:</Subscript> effect of water addition and vacuum condition

DHA/EPA-rich phosphatidylcholine (PC) was successfully synthesized by immobilized phospholipase A₁ (PLA₁)-catalyzed transesterification of PC and DHA/EPA-rich ethyl esters in a solvent-free system. Effects of reaction temperature, water addition and substrate mass ratio on the incorporation of DHA/EPA were evaluated using response surface methods (RSM). Water addition had most significant effect on the incorporation. Reaction temperature and substrate mass ratio, however, had no significant effect on the incorporation. The maximal incorporation was 19.09 % (24 h) under the following conditions: temperature 55.7 °C, water addition 1.1 wt % and substrate mass ratio (ethyl esters/PC) 6.8:1. Furthermore, effects of water addition (from 0 to 1.25 wt %) on DHA/EPA incorporation and the composition of products were further investigated. The immobilized PLA₁ was more active when water addition was above 0.5 wt %. By monitoring the reaction processes with different water addition, a possible reaction scheme was proposed for transesterification of PC with DHA/EPA-rich ethyl esters. In summary, PC and sn2-lysophosphatidylocholine (LPC) were predominant in the mixtures at early stages of reaction, whereas sn1-LPC and glycerophosphocholine (GPC) predominant at later stages. The vacuum employed after 24 h significantly increased the incorporation of DHA/EPA and the composition of PC, and the highest incorporation (30.31 %) of DHA/EPA was obtained at 72 h and the yield of PC was 47.2 %.     

Membrane microreactor in biocatalytic transesterification of triolein for biodiesel production

Transesterification is a principal chemical reaction that occurs in biodiesel production. We developed a novel biocatalytic membrane microreactor (BMM) for continuous transesterification by utilizing an asymmetric membrane as an enzyme-carrier for immobilization. The BMM was developed by pressure driven filtration of lipase from Pseudomonas fluorescens, which is suitable for highly efficient biocatalytic transesterification. Lipase solution was allowed to permeate through an asymmetric membrane with NMWL 300 kDa composed of polyethersulfone. The performances of BMM were studied in biodiesel synthesis via transesterification of triolein with methanol. Transesterification was carried out by passing a solution of triolein and methanol through the asymmetric membrane. The degree of triolein conversion using this microreactor was ca. 80% with a reaction time of 19 min. The BMM system displayed good stability, with no activity decay over a period of 12 day with continuous operation. Results from triolein transesterification clearly demonstrate the potential of an asymmetric membrane as an enzyme carrier material. Enzyme activity (mmol/h·g_lipase) was approximately 3 fold higher than that of native free lipase.     

Enzymatic hydrolysis of steryl glucosides,major contaminants of vegetable oil-derived biodiesel

Biodiesels are mostly produced from lipid transesterification of vegetable oils, including those from soybean, jatropha, palm, rapeseed, sunflower, and others. Unfortunately, transesterification of oil produces various unwanted side products, including steryl glucosides (SG), which precipitate and need to be removed to avoid clogging of filters and engine failures. So far, efficient and cost-effective methods to remove SGs from biodiesel are not available. Here we describe for the first time the identification, characterization and heterologous production of an enzyme capable of hydrolyzing SGs. A synthetic codon-optimized version of the lacS gene from Sulfolobus solfataricus was efficiently expressed and purified from Escherichia coli, and used to treat soybean derived biodiesel containing 100 ppm of SGs. After optimizing different variables, we found that at pH 5.5 and 87 °C, and in the presence of 0.9 % of the emulsifier polyglycerol polyricinoleate, 81 % of the total amount of SGs present in biodiesel were hydrolyzed by the enzyme. This remarkable reduction in SGs suggests a path for the removal of these contaminants from biodiesel on industrial scale using an environmentally friendly enzymatic process.     

Application of multi-component reaction for covalent immobilization of two lipases on aldehyde-functionalized magnetic nanoparticles; production of biodiesel from waste cooking oil

Lipase from Rhizomucor miehei (RML) and Thermomyces lanuginosa lipase (TLL) were immobilized on silica core-shell magnetic nanoparticles (Fe₃O₄@SiO₂) produced by coating Fe₃O₄ core with silica shell. The nanoparticles were functionalized with aldehyde groups followed by immobilization of RML and TLL by using a multi-component reaction in an extremely mild condition. Rapid immobilization of both enzymes (1.5−12 h) with high immobilization yields (81–100%) was observed. The maximum loading capacity of the support was determined to be 81 mg for RML and 97 mg for TLL. The thermal stability of the immobilized derivatives of RML and TLL were greatly improved by retaining 54 and 97 % of their initial activities at 65 °C, respectively. The immobilized preparations were used to produce biodiesel by transesterification of waste cooking oil. In an optimization study, Response Surface Methodology (RSM) and a central composite rotatable design (CCRD) were used to study the effect of amount of biocatalyst, temperature, reaction time, water adsorbent (wt.%) and ratio of t-butanol to oil (wt.%) on the yield of biodiesel production. Biodiesel production yield by immobilized TLL reached 93.1 % under optimal conditions while the maximum yield for RML was 57.5 %. Both immobilized derivatives showed high reusability after 5 cycles of the reaction.