Optimization of lipase-catalyzed biodiesel by response surface methodology

Biodiesel prepared by catalyzed mild transesterification has become of much current interest for bioenergy. The ability of a commercial immobilized lipase (Novo Industries--Bagsvaerd, Denmark) from Rhizomucor miehei (Lipozyme IM-77) to catalyze the transesterification of soybean oil and methanol was investigated in this study. Response surface methodology and 5-level-5-factor central composite rotatable design were employed to evaluate the effects on reaction time, temperature, enzyme amount, molar ratio of methanol to soybean oil, and added water content on percentage weight conversion to soybean oil methyl ester by transesterification. Based on ridge max analysis, the optimum synthesis conditions giving 92.2% weight conversion were: reaction time 6.3 h, temperature 36.5 degrees C, enzyme amount 0.9 BAUN (Batch Acidolysis Units NOVO), substrate molar ratio 3.4:1, and added water 5.8%.     

Direct preparation of biodiesel from rapeseed oil leached by two-phase solvent extraction

A new method which coupled the two-phase solvent extraction (TSE) with the synthesis of biodiesel was studied. Investigations were carried out on transesterification of methanol with oil-hexane solution coming from TSE process in the presence of sodium hydroxide as the catalyst. Biodiesel (fatty acid methyl esters) were the products of transesterification. The influential factors of transesterification, such as reaction time, catalyst concentration, mole ratio of methanol to oil and reaction temperature were optimized. The results showed that the optimal reaction parameters were sodium hydroxide concentration 1.1% by weight of rapeseed oil, mole ratio of methanol to oil 9:1, reaction time 120 min, and reaction temperature 55-60 degrees C. Under these conditions, the TG conversion would rise up to 98.2%. Based on the new method, biodiesel production process could be simplified and the biodiesel cost could be reduced.     

In situ alkaline transesterification of cottonseed oil for production of biodiesel and nontoxic cottonseed meal

The production of fatty acid methyl ester (FAME) by direct in situ alkaline-catalyzed transesterification of the triglycerides (TG) in cottonseeds was examined. The experimental results showed that the amount of cottonseed oil dissolved in methanol was approximately 99% of the total oil and the conversion of this oil could achieve 98% under the following conditions: less than 2% moisture content in cottonseed flours, 0.3-0.335mm particle size, 0.1mol/L NaOH concentration in methanol, 135:1 methanol/oil mole ratio, 40 degrees C reaction temperature and 3h reaction time. Further, the effects of co-solvent petroleum ether and methanol recycling on the cottonseed oil extraction and conversion were also investigated. The use of alkaline methanol as extraction and reaction solvent, which would be useful for extraction oil and gossypol, would reduce the gossypol content in the cottonseed meal. The free and total gossypol contents in the cottonseed meal obtained from in situ alkaline transesterification were far below the FAO standard. And the nontoxic cottonseed meal could be used as animal protein feed resources.     

Production of biodiesel and lactic acid from rapeseed oil using sodium silicate as catalyst

Biodiesel and lactic acid from rapeseed oil was produced using sodium silicate as catalyst. The transesterification in the presence of the catalyst proceeded with a maximum yield of 99.6% under optimized conditions [3% (w/w) sodium silicate, methanol/oil molar ratio 9/1, reaction time 60 min, reaction temperature 60 °C, and stirring rate 250 rpm]. After six consecutive transesterification reactions, the catalyst was collected and used for catalysis of the conversion of glycerol to lactic acid. A maximum yield of 80.5% was achieved when the reaction was carried out at a temperature of 300 °C for 90 min. Thus, sodium silicate is an effective catalyst for transesterification and lactic acid production from the biodiesel by-product, glycerol.     

Lipase-catalysed production of biodiesel fuel from some Nigerian lauric oils

Fatty acids esters were produced from two Nigerian lauric oils, palm kernel oil and coconut oil, by transesterification of the oils with different alcohols using PS30 lipase as a catalyst. In the conversion of palm kernel oil to alkyl esters (biodiesel), ethanol gave the highest conversion of 72%, t-butanol 62%, 1-butanol 42%, n-propanol 42% and iso-propanol 24%, while only 15% methyl ester was observed with methanol. With coconut oil, 1-butanol and iso-butanol achieved 40% conversion, 1-propanol 16% and ethanol 35%, while only traces of methyl esters were observed using methanol. Studies on some fuel properties of palm kernel oil and its biodiesel showed that palm kernel oil had a viscosity of 32.40 mm2/s, a cloud point of 28 degrees C and a pour point of 22 degrees C, while its biodiesel fuel had a viscosity of 9.33 mm2/s, a cloud point of 12 degrees C and a pour point of 8 degrees C. Coconut oil had a viscosity of 28.58 mm(2)/s, a cloud point of 27 degrees C and a pour point of 20 degrees C, while its biodiesel fuel had a viscosity of 7.34 mm2/s, a cloud point of 5 degrees C and a pour point of -8 degrees C. Some of the fuel properties compared favourably with international biodiesel specifications.     

Acceleration of catalytic activity of calcium oxide for biodiesel production

This research was aimed at studying the acceleration of the catalytic activity of calcium oxide (CaO) for developing an effective heterogeneous catalyst for biodiesel production by the transesterification of plant oil with methanol. CaO was activated by pretreatment with methanol and was used for the transesterification reaction. The activation and transesterification reaction conditions were examined. The obtained optimal reaction conditions were 0.1-g CaO, 3.9-g methanol, 15-g rapeseed oil, and 1.5-h activation time at room temperature that provided methyl ester in approximately 90% yield within a reaction time of 3h at 60 degrees C. The activation mechanism was also investigated, and the proposed mechanism is as follows. By pretreatment with methanol, a small amount of CaO gets converted into Ca(OCH(3))(2) that acts as an initiating reagent for the transesterification reaction and produces glycerin as a by-product. Subsequently, a calcium-glycerin complex, formed due to the reaction of CaO with glycerin, functions as the main catalyst and accelerates the transesterification reaction.     

Biodiesel production from heterotrophic microalgal oil

The present study introduced an integrated method for the production of biodiesel from microalgal oil. Heterotrophic growth of Chlorella protothecoides resulted in the accumulation of high lipid content (55%) in cells. Large amount of microalgal oil was efficiently extracted from these heterotrophic cells by using n-hexane. Biodiesel comparable to conventional diesel was obtained from heterotrophic microalgal oil by acidic transesterification. The best process combination was 100% catalyst quantity (based on oil weight) with 56:1 molar ratio of methanol to oil at temperature of 30 degrees C, which reduced product specific gravity from an initial value of 0.912 to a final value of 0.8637 in about 4h of reaction time. The results suggested that the new process, which combined bioengineering and transesterification, was a feasible and effective method for the production of high quality biodiesel from microalgal oil.     

Preparation,application, and optimization of Zn/Al complex oxides for biodiesel production under sub-critical conditions

Biodiesel produced by transesterification is a promising green fuel in the future. A new heterogeneous catalyst, Zn/Al complex oxide, was prepared for biodiesel production. The results showed that the catalyst derived from a hydrotalcite-like precursor with a zinc/aluminum atom ratio of 3.74:1 and calcined at 450 °C gave the highest conversion of 84.25%. Analysis of XRD, XPS, FI-IF, TG-DTA, BET and alkalinity tests demonstrated that it is the unique structure of hydrotalcite-like compound precursor that gave the catalyst a high alkalinity greater than 11.1. The optimal reaction condition for Zn/Al complex oxide was under methanol sub-critical condition: 200 °C, 2.5 MPa, 1.4% (wt) catalyst dosage, and 24:1 methanol to oil ratio. Under these conditions, the conversion reached 84.25% after 90 min, which was better than Mg/Al complex oxides. The excellent tolerance to water and free fatty acid was exhibited when the oil feed had fewer than 6% FFA or 10% water content with a conversion greater than 80%.     

Lipase catalyzed methanolysis to produce biodiesel: Optimization of the biodiesel production

A lipase from Candida sp., suitable for transesterification of fats and oils to produce fatty acid methyl ester (FAME), was immobilized on a cheap cotton membrane, in this paper. The conversion ratio of salad oil to biodiesel could reach up to 96% with the optimal reaction conditions. Continuous reaction in a fixed bed reactor was also investigated. A three-step transesterification with methanol (methanolysis) of oil was conducted by using a series of nine columns packed with immobilized Candida sp. 99–125 lipase. As substrate of the first reaction step, plant or waste oil was used together with 1/3 molar equivalent of methanol against total fatty acids in the oil. Mixtures of the first- and second-step eluates and 1/3 molar equivalent of methanol were used for the second- and third-reaction steps. A hydrocyclone was used in order to on-line separate the by-product glycerol after every 1/3 molar equivalent of methanol was added. Petroleum ether was used as solvent (3/2, v/v of oil) and the pump was operated with a flow rate of 15 L/h giving an annual throughput of 100 t. The final conversion ratio of the FAME from plant oil and waste oil under the optimal condition was 90% and 92%, respectively. The life of the immobilized lipase was more than 10 days. This new technique has many strongpoints such as low pollution, environmentally friendly, and low energy costs.     

Development of an amphiphilic matrix for immobilization of <Emphasis Type="Italic">Candida antartica</Emphasis> lipase B for biodiesel production

Biodiesel has been greatly interested as an alternative fuel and is produced by a transesterification reaction of oil with alcohol. Recently, microbial lipases have been used for biodiesel production. Among the microbial lipase, immobilized Candida antartica lipase B (CALB) is the most widely used. However, CALB is unstable and shows low catalytic efficiency in the reaction media because the reaction media contains a high concentration of methanol and the lipase is also inhibited by the by-product glycerol. In this study, to overcome these limitations, we developed an amphiphilic matrix to immobilize CALB. The immobilized lipase in an amphiphilic matrix with 80% ethyltrimethoxysilane (ETMS) in tetramethoxysilane (TMOS) and pretreated with oil showed the highest specific activity and biodiesel conversion ratio; about 90% biodiesel conversion in 24 h at an initial molar ratio of 1: 1 (oil: methanol) with stepwise methanol feeding in order to adjust the net molar ratio to be 1: 3.     

Influence of fatty acid composition of raw materials on biodiesel properties

The aim of this work was the study of the influence of the raw material composition on biodiesel quality, using a transesterification reaction. Thus, ten refined vegetable oils were transesterificated using potassium methoxide as catalyst and standard reaction conditions (reaction time, 1h; weight of catalyst, 1 wt.% of initial oil weight; molar ratio methanol/oil, 6/1; reaction temperature, 60 degrees C). Biodiesel quality was tested according to the standard [UNE-EN 14214, 2003. Automotive fuels. Fatty acid methyl esters (FAME) for diesel engines. Requirements and test methods]. Some critical parameters like oxidation stability, cetane number, iodine value and cold filter plugging point were correlated with the methyl ester composition of each biodiesel, according to two parameters: degree of unsaturation and long chain saturated factor. Finally, a triangular graph based on the composition in monounsaturated, polyunsaturated and saturated methyl esters was built in order to predict the critical parameters of European standard for whatever biodiesel, known its composition.     

Development of heterogeneous base catalysts for biodiesel production

Investigations were conducted on heterogeneous base catalysts for the transesterification of oil aimed at effective production of biodiesel. Thirteen different kinds of metal oxides containing calcium, barium, magnesium, or lanthanum were prepared as catalysts. Their catalytic activities were tested for transesterification at 60 degrees C with a 6:1 molar ratio of methanol to oil and a reaction time of 10h. The calcium-containing catalysts - CaTiO3, CaMnO3, Ca2Fe2O5, CaZrO3, and CaO-CeO2 - showed high activities and approximately 90% yields of methyl ester. Furthermore, catalytic durability tests were performed by repeating the transesterification reaction several times with the calcium-containing catalysts recovered from the previous reaction mixture. It was found that CaZrO3 and CaO-CeO2 show high durability and have the potential to be used in biodiesel production processes as heterogeneous base catalysts.     

Lipase-catalyzed biodiesel production with methyl acetate as acyl acceptor

Biodiesel is an alternative diesel fuel made from renewable biological resources. During the process of biodiesel production, lipase-catalyzed transesterification is a crucial step. However, current techniques using methanol as acyl acceptor have lower enzymatic activity; this limits the application of such techniques in large-scale biodiesel production. Furthermore, the lipid feedstock of currently available techniques is limited. In this paper, the technique of lipase-catalyzed transesterification of five different oils for biodiesel production with methyl acetate as acyl acceptor was investigated, and the transesterification reaction conditions were optimized. The operation stability of lipase under the obtained optimal conditions was further examined. The results showed that under optimal transesterification conditions, both plant oils and animal fats led to high yields of methyl ester: cotton-seed oil, 98%; rapeseed oil, 95%; soybean oil, 91%; tea-seed oil, 92%; and lard, 95%. Crude and refined cottonseed oil or lard made no significant difference in yields of methyl ester. No loss of enzymatic activity was detected for lipase after being repeatedly used for 40 cycles (ca. 800 h), which indicates that the operational stability of lipase was fairly good under these conditions. Our results suggest that cotton-seed oil, rape-seed oil and lard might substitute soybean oil as suitable lipid feedstock for biodiesel production. Our results also show that our technique is fit for various lipid feedstocks both from plants and animals, and presents a very promising way for the large-scale biodiesel production.     

Transesterification of palm oil to methyl ester on activated carbon supported calcium oxide catalyst

In this study, methyl ester (ME) was produced by transesterification of palm oil (CPO) (cooking grade) using activated carbon supported calcium oxide as a solid base catalyst (CaO/AC). Response surface methodology (RSM) based on central composite design (CCD) was used to optimize the effect of reaction time, molar ratio of methanol to oil, reaction temperature and catalyst amount on the transesterification process. The optimum condition for CPO transesterification to methyl ester was obtained at 5.5 wt.% catalyst amount, 190 °C temperature, 15:1 methanol to oil molar ratio and 1 h 21 min reaction time. At the optimum condition, the ME content was 80.98%, which is well within the predicted value of the model. Catalyst regeneration studies indicate that the catalyst performance is sustained after two cycles.     

The effect of the acidity of rapeseed oil on its transesterification

The aim of this work is to study the transesterification of vegetable oil with a high acid number at unchanged reaction conditions. Rapeseed oil was used as the raw material and its acid number was changed by the addition of oleic acid (from 0.89 to 12.25 mg KOH/g). Methanol was used for transesterification (molar ratio of oil to methanol 1:6) and potassium hydroxide was used as a catalyst. After the reaction time, the residue of the catalyst was neutralised by gaseous carbon dioxide and the methanol excess was removed. After the separation of two phases, each of them was analyzed (in the ester phase: yield, content of methyl ester and acid number; in the glycerol phase: yield, density, viscosity, content of glycerol, soaps, methyl ester, potassium carbonate and hydrogen carbonate). The obtained data was compared with theoretical material balances and the effect on the saponification of oil was discussed. The results show that the yield of methyl ester (biodiesel) is significantly affected by a higher acid number, as well as enhanced soap formation. On the other hand, the conversion of the oil and acid number of the ester phase remain at constant values in studied borders.     

Biodiesel production from high acid value waste frying oil catalyzed by superacid heteropolyacid

Transesterification of waste cooking oil with high acid value and high water contents using heteropolyacid H3PW12O40 x 6H2O (PW12) as catalyst was investigated. The hexahydrate form of PW(12) was found to be the most promising catalyst which exhibited highest ester yield 87% for transesterification of waste cooking oil and ester yield 97% for esterification of long-chain palmitic acid, respectively. The PW12 acid catalyst shows higher activity under the optimized reaction conditions compared with conventional homogeneous catalyst sulfuric acid, and can easily be separated from the products by distillation of the excess methanol and can be reused more times. The most important feature of this catalyst is that the catalytic activity is not affected by the content of free fatty acids (FFAs) and the content of water in the waste cooking oil and the transesterification can occur at a lower temperature (65 degrees C), a lower methanol oil ratio (70:1) and be finished within a shorter time. The results illustrate that PW12 acid is an excellent water-tolerant and environmentally benign acid catalyst for production of biodiesel from waste cooking oil.     

Lipase-mediated Transformation of Vegetable Oils into Biodiesel using Propan-2-ol as Acyl Acceptor

Propan-2-ol was used as an acyl acceptor for immobilized lipase-catalyzed preparation of biodiesel. The optimum conditions for transesterification of crude jatropha (Jatropha curcas), karanj (Pongamia pinnata) and sunflower (Helianthus annuus) oils were 10% Novozym-435 (immobilized Candida antarctica lipase B) based on oil weight, alcohol to oil molar ratio of 4:1 at 50 °C for 8 h. The maximum conversions achieved using propan-2-ol were 92.8, 91.7 and 93.4% from crude jatropha, karanj and sunflower oils, respectively. Reusability of the lipase was maintained over 12 repeated cycles with propan-2-ol while it reached to zero by 7^th cycle when methanol was used as an acyl acceptor, under standard reaction conditions. Revisions requested 22 December 2005; Revisions received 26 January 2006     

Enzymatic transesterification of olive oil and its precursors

The effect of different solvents and three different acyl acceptors on the transesterification of triolein (as a model compound) was investigated. The yield of biodiesel (methyl or ethyl ester) was monitored as a function of time. The yield of the product was also determined in a solvent-free system for two different modes of stirring. The results indicate that the highest yield is obtained in a solvent-free system with mechanical stirring. Methyl acetate is also effective as a solvent and acyl acceptor. Biodiesel was also produced by transesterification of triglycerides (triolein) present in olive oil with methanol and Novozym® 435. The effect of the molar ratio of methanol to triolein, mode of methanol addition, enzyme activity and reaction temperature on overall conversion and yield was determined. The final conversion and yield of biodiesel after a reaction time of 24 h were unaffected by changes in these parameters over the range studied. Preliminary findings indicate that the results obtained from small scale reactors and fresh oil can be extended to larger reactors and used oil.     

Soybean oil methyl esters preparation using NaX zeolites loaded with KOH as a heterogeneous catalyst

The transesterification of soybean oil with methanol to methyl esters was carried out using NaX zeolites loaded with KOH as a solid base catalyst. Best result was obtained with NaX zeolite loaded with 10% KOH, followed by heating at 393 K for 3 h. When the transesterification reaction was carried out at reflux of methanol (338 K), with a 10:1 molar ratio of methanol to soybean oil, a reaction time of 8 h and a catalyst amount of 3 wt.%, the conversion of soybean oil was 85.6%.     

Enzymatic transesterification of olive oil and its precursors

The effect of different solvents and three different acyl acceptors on the transesterification of triolein (as a model compound) was investigated. The yield of biodiesel (methyl or ethyl ester) was monitored as a function of time. The yield of the product was also determined in a solvent-free system for two different modes of stirring. The results indicate that the highest yield is obtained in a solvent-free system with mechanical stirring. Methyl acetate is also effective as a solvent and acyl acceptor. Biodiesel was also produced by transesterification of triglycerides (triolein) present in olive oil with methanol and Novozym® 435. The effect of the molar ratio of methanol to triolein, mode of methanol addition, enzyme activity and reaction temperature on overall conversion and yield was determined. The final conversion and yield of biodiesel after a reaction time of 24 h were unaffected by changes in these parameters over the range studied. Preliminary findings indicate that the results obtained from small scale reactors and fresh oil can be extended to larger reactors and used oil.