Biodiesel production using enzymatic transesterification - Current state and perspectives

Biodiesel has attracted considerable interest in recent years as an alternative, biodegradable and nonpolluting transportation fuel. Conventional alkaline process for biodiesel production are energy-consuming and generate undesirable by-products such as soaps, that make difficult the separation and purification of biodiesel.Particular attention has been dedicated to the use of lipases as biocatalysts for biodiesel production due to their favorable conversion rate obtained in gentle conditions and relatively simple downstream processing steps for the purification of biodiesel and by-products. However, comparatively to conventional chemical processes, the major obstacles for enzymatic production of biodiesel remain the cost of lipases, the relatively slower reaction rate and lipases inactivation caused by methanol and glycerol.This review evaluates the current status and perspectives for enzymatic biodiesel production and indicates the key operational variables that influence lipase activity and stability together with the technological solutions for industrial implementation of enzymatic process.     

Biodiesel production from mixed soybean oil and rapeseed oil

The biodiesel (fatty acid methyl esters, FAME) was prepared by transesterification of the mixed oil (soybean oil and rapeseed oil) with sodium hydroxide (NaOH) as catalyst. The effects of mole ratio of methanol to oil, reaction temperature, catalyst amount and reaction time on the yield were studied. In order to decrease the operational temperature, a co-solvent (hexane) was added into the reactants and the conversion efficiency of the reaction was improved. The optimal reaction conditions were obtained by this experiment: methanol/oil mole ratio 5.0:1, reaction temperature 55 °C, catalyst amount 0.8 wt.% and reaction time 2.0 h. Under the optimum conditions, a 94% yield of methyl esters was reached ∼94%. The structure of the biodiesel was characterized by FT-IR spectroscopy. The sulfur content of biodiesel was determined by Inductively Coupled Plasma emission spectrometer (ICP), and the satisfied result was obtained. The properties of obtained biodiesel from mixed oil are close to commercial diesel fuel and is rated as a realistic fuel as an alternative to diesel. Production of biodiesel has positive impact on the utilization of agricultural and forestry products.     

Biodiesel production from castor oil in Brazil: A difficult reality

The Brazilian National Program for Production and Use of Biodiesel (PNPB in Portuguese) has created a huge demand for biodiesel in Brazil. The PNPB is strongly based on social development through the inclusion of family farmers in projects integrated with biodiesel power plants. Among the various oilseeds, castor bean (Ricinus communis L.) was identified as the ideal one to promote social development in the semi-arid region. However, although promising, the mechanisms of the federal program are still insufficient to promote the effective participation of family farmers. This research shows that companies are facing huge problems in implementing contracts with family farmers. It describes and analyzes the functioning dynamics of this agro-production chain. This paper addresses the identification and the discussion of these obstacles, in order to increase the competitiveness of the biodiesel agribusiness chain, based on castor oil social projects in Brazil.     

Biodiesel production from waste cooking oil over sulfonated catalysts

Biodiesel production from waste cooking oil with methanol was carried out in the presence of poly(vinyl alcohol) with sulfonic acid groups (PVA-SO₃H) and polystyrene with sulfonic acid groups (PS-SO₃H), at 60°C. The PVA-SO₃H catalyst showed higher catalytic activity than the PS-SO₃H one. In order to optimize the reaction conditions, different parameters were studied. An increase of waste cooking oil conversion into fatty acid methyl esters with the amount of PVA-SO₃H was observed. When the transesterification and esterification of WCO was carried out with ethanol over PVA-SO₃H, at 60°C, a decrease of biodiesel production was also observed. The WCO conversion into fatty acid ethyl ester increased when the temperature was increased from 60 to 80°C. When different amounts of free fatty acids were added to the reaction mixture, a slight increase on the conversion was observed. The PVA-SO₃H catalyst was reused and recycled with negligible loss in the activity.     

Biodiesel production from crude rice bran oil and properties as fuel

This research reported on the successfully production of biodiesel by transesterification of crude rice bran oil (RBO). The process included three-steps. Firstly, the acid value of RBO was reduced to below 1 mg KOH/g by two-steps pretreatment process in the presence of sulfuric acid catalyst. Secondly, the product prepared from the first process was carried out esterification with an alkaline catalyst. The influence of four variables on conversion efficiency to methyl ester, i.e., methanol/RBO molar ratio, catalyst amount, reaction temperature and reaction time, was studied at this stage. The content of methyl ester was analyzed by chromatographic analysis. Through orthogonal analysis of parameters in a four-factor and three-level test, the optimum reaction conditions for the transesterification were obtained: methanol/RBO molar ratio 6:1, usage amount of KOH 0.9% w/w, reaction temperature 60 °C and reaction time 60 min. In the third step, methyl ester prepared from the second processing step was refined to become biodiesel. Fuel properties of RBO biodiesel were studied and compared according to ASTM D6751-02 and DIN V51606 standards for biodiesel. Most fuel properties complied with the limits prescribed in the aforementioned standards. The consequent engine test showed a similar power output compared with regular diesel but consumption rate was slightly higher. Emission tests showed a marked decrease in CO, HC and PM, however, with a slight increase in NO_X.     

Biodiesel production from olive–pomace oil of steam-treated alperujo

Recently interest has been revived in the use of plant-derived waste oils as renewable replacements for fossil diesel fuel. Olive–pomace oil (OPO) extracted from alperujo (by-product of processed olives for olive oil extraction), and produced it in considerable quantities throughout the Mediterranean countries, can be used for biodiesel production. A steam treatment of alperujo is being implemented in OPO extraction industry. This steam treatment improves the solid–liquid separation by centrifugation and facilitates the drying for further extraction of OPO. It has been verified that the steam treatment of this by-product also increases the concentration of OPO in the resulting treated solid, a key factor from an economic point of view. In the present work, crude OPO from steam-treated alperujo was found to be good source for producing biodiesel. Oil enrichment, acidity, biodiesel yield and fatty acid methyl ester composition were evaluated and compared with the results of the untreated samples. Yields and some general physicochemical properties of the quality of biodiesel were also compared to those obtained with other oils commonly used in biodiesel production. As for biodiesel yield no differences were observed. A transesterification process which included two steps was used (acid esterification followed by alkali transesterification). The maximum biodiesel yield was obtained using molar ratio methanol/triglycerides 6:1 in presence of sodium hydroxide at a concentration of 1% (w/w), reaction temperature 60 °C and reaction time 80 min. Under these conditions the process gave yields of about 95%, of the same order as other feedstock using similar production conditions.     

Biodiesel from different oil using fixed-bed and plug-flow reactors

In this study, a pilot scale of 100 t/year biodiesel production system, mainly consisting of a fixed-bed and a down-stream plug-flow reactors, was setup to test different feedstock oils, especially a kind of high-acidified oil, trap grease, for their feasibility as biodiesel feedstock in China. The tested oils include three kinds of typical oil from Guangdong Province, China: rapeseed oil, Chinese wood oil, and trap grease. At the same time the optimum residence time for a plug-flow reactor to perform transesterification reaction was investigated in this study. At the temperature of 65 °C, methanol/oil molar ratio of 6:1 and KOH load of 1.2 wt% of oil, the optimum residence time was found to be 19 min. A type of ion-exchange resin was used to fill in the fixed-bed reactor and used as the esterification catalyst for pretreating on the high-acidified oil. For the fresh catalyst, the acid value of trap grease could be reduced from 114 mg KOH/g to about 2 mg KOH/g after 13 h at temperature 75 °C, catalyst load of 15 wt% of oil, methanol addition of 20 wt% of oil. The lifetime test for the catalyst indicated that its life is over 30 days. The quality of biodiesel derived from three feedstock oils is compared with newly published China BD100 standard of GB/T20828-2007. A comparison of the results reveals that the biodiesel generated through this system could satisfactorily meet China BD100 standard. It indicates that the designed process in this system has a good adaptability for different kinds of oil.     

Biodiesel production from refined sunflower vegetable oil over KOH/ZSM5 catalysts

ZSM5 zeolite was impregnated with different KOH loadings (15 wt.%, 25 wt.% and 35 wt.%) to prepare a series of KOH/ZSM5 catalysts. The catalysts were calcined at 500 °C for 3 h and then characterized by N₂ adsorption–desorption and X-ray diffraction (XRD) techniques. The catalysts were tested in the transesterification reaction in a batch reactor at 60 °C and under atmospheric pressure. It was found that KOH/ZSM5 with 35 wt.% loading showed the best catalytic performance. The best reaction conditions in the presence of KOH/ZSM5 (35 wt.%) were determined while modifying the catalyst to oil ratio and the reaction time. The highest methyl ester yield (>95%) was obtained for a reaction time of 24 h, a catalyst to oil ratio of 18 wt.%, and a methanol to oil molar ratio of 12:1. The properties of produced biodiesel complied with the ASTM specifications. The catalytic stability test showed that 35KOH/ZSM5 was stable for 3 consecutive runs. Characterization of the spent catalyst indicated that a slight deactivation might be due to the leaching of potassium oxides active sites.     

Lipase immobilization and production of fatty acid methyl esters from canola oil using immobilized lipase

Lipase enzyme from Aspergillus oryzae (EC 3.1.1.3) was immobilized onto a micro porous polymeric matrix which contains aldehyde functional groups and methyl esters of long chain fatty acids (biodiesel) were synthesized by transesterification of crude canola oil using immobilized lipase. Micro porous polymeric matrix was synthesized from styrene-divinylbenzene (STY-DVB) copolymers by using high internal phase emulsion technique and two different lipases, Lipozyme TL-100L^® and Novozym 388^®, were used for immobilization by both physical adsorption and covalent attachment. Biodiesel production was carried out with semi-continuous operation. Methanol was added into the reactor by three successive additions of 1:4 M equivalent of methanol to avoid enzyme inhibition. The transesterification reaction conditions were as follows: oil/alcohol molar ratio 1:4; temperature 40 °C and total reaction time 6 h. Lipozyme TL-100L^® lipase provided the highest yield of fatty acid methyl esters as 92%. Operational stability was determined with immobilized lipase and it indicated that a small enzyme deactivation occurred after used repeatedly for 10 consecutive batches with each of 24 h. Since the process is yet effective and enzyme does not leak out from the polymer, the method can be proposed for industrial applications.     

Biodiesel production from Cedrus deodara oil in different types of ultrasonic reactors and energy analysis

This paper deals with the production of biodiesel using vegetable oil, extracted from Deodar (Cedrus deodara) in various types of ultrasonic reactors. The biodiesel so produced is tested for its property and stability. Biodiesel yield is optimized as a function of reaction time for various ultrasonic reactors. The biodiesel production through the triple-frequency flow cell ultrasonic reactor is found the most energy efficient when compared to other types of ultrasonic reactors. Biodiesel so produced from deodar oil is stable under atmospheric conditions with its various physicochemical properties within the range of acceptable limits of the diesel engine.     

Biodiesel production from waste cooking oil using heterogeneous catalysts and its operational characteristics on variable compression ratio CI engine

In the present work, the optimum biodiesel conversion from waste cooking oil to biodiesel through transesterification method was investigated. The base catalyzed transesterification under different reactant proportions such as the molar ratio of alcohol to oil and mass ratio of catalyst to oil was studied for optimum production of biodiesel. The optimum condition for base catalyzed transesterification of waste cooking oil was determined to be 12:1 and 5 wt% of zinc doped calcium oxide. The fuel properties of the produced biodiesel such as the calorific value, flash point and density were examined and compared to conventional diesel. The properties of produced biodiesel and their blend for different ratios (B20, B40, B60, B80 and B100) were comparable with properties of diesel oil and ASTM biodiesel standards. Tests have been conducted on CI engine which runs at a constant speed of 1500 rpm, injection pressure of 200 bar, compression ratio 15:1 and 17.5, and varying engine load. The performance parameters include brake thermal efficiency, brake specific energy consumption and emissions parameters such as Carbon monoxide (CO), Hydrocarbon (HC), Oxides of Nitrogen (NOx) and smoke opacity varying with engine load (BP). Diesel engine's thermal performance and emission parameters such as CO, HC, and NOx on different biodiesel blends demonstrate that biodiesel produced from waste cooking oil using heterogeneous catalyst was suitable to be used as diesel oil blends and had lesser emissions as compared to conventional diesel.     

The influence of free fatty acid intermediate on biodiesel production from soybean oil by whole cell biocatalyst

The whole cell of lipase-producing Rhizopus oryzae was employed as biocatalyst for transesterification of soybean oil containing oleic acid. The free fatty acid (FFA) intermediate, playing an important role in the kinetics of transesterification of soybean oil, was thoroughly investigated and characterized. The conversion was more than 97% at the initial FFA content of 5.5%. A high content of FFA could protect the lipase from denaturation. The 34.6 percent of FFA with the optimal 26-mg mL⁻¹ methanol resulted in a specific reaction rate of 420 mg h⁻¹g-dry cell⁻¹. In addition, the methanol/FFA ratio at 0.83-1.7 provides a good indication of the fatty acid methyl esters conversions for different initial FFA contents. In the transesterification process, more FFA intermediate present would become beneficial to conversion of retrograde feedstock to biodiesel. The immediately generated and original FFA content become the major rate-determining factor in the FFA-mixed transesterification process.     

Biodiesel production from Nerium oleander (Thevetia peruviana) oil through conventional and ultrasonic irradiation methods

In the present research work, Nerium oleander oil has been used as raw material for producing biodiesel using both ultrasonic transesterification and a magnetic stirrer method. A two-step transesterification process was carried out for optimum condition of 0.40% V/V methanol to oil ratio, 1% V/V H₂SO₄ catalyst, 55°C temperature, and 60 min reaction time followed by treatment with 0.2% V/V methanol to oil ratio, 1% V/W KOH alkaline catalyst, 55°C temperature, and 60 min reaction time. The process is repeated with an ultrasonic method at the frequency of 28 kHz using ultrasonic horn type reactor (50 W) for about 10–15 min. Biodiesel obtained from ultrasonic method and magnetic stirrer was then compared for their percentage yield and physiochemical properties. Ultrasonic transesterification process gave a maximum yield of 97% by weight of oleander biodiesel along with improved physiochemical characteristics. Therefore, it is concluded that ultrasonic method is the most effective method for converting crude oleander oil into biodiesel.     

Accurate modeling of biodiesel production from castor oil using ANFIS

Research for finding alternative fuel sources has been concluded that the renewable fuels such as biodiesel can be used as an alternative to fossil fuels because of the energy security reasons and environmental benefits. In this contribution, transesterification of castor oil with methanol to form biodiesel has been modeled by using artificial neural network fuzzy interference system (ANFIS) approach. Methanol to oil molar ratio, catalyst amount (C), temperature (T), and time (S) were used as input parameters and fatty acid methyl ester yield was used as output parameter for modeling the efficiency of biodiesel production from castor oil. Obtaining low value of absolute deviation (2.2391), high value of R-squared (0.98704), and other modeling results proves that ANFIS modeling is an effective approach for biodiesel production from castor oil. In conclusion, comparison between our model and other previous predictive models reported in open literature indicates the priority of our model.     

Optimization of biodiesel production from camelina oil using orthogonal experiment

Camelina oil is a low-cost feedstock for biodiesel production that has received a great deal of attention in recent years. This paper describes an optimization study on the production of biodiesel from camelina seed oil using alkaline transesterification. The optimization was based on sixteen well-planned orthogonal experiments (OA₁₆ matrix). Four main process conditions in the transesterification reaction for obtaining the maximum biodiesel production yield (i.e. methanol quantity, reaction time, reaction temperature and catalyst concentration) were investigated. It was found that the order of significant factors for biodiesel production is catalyst concentration > reaction time > reaction temperature > methanol to oil ratio. Based on the results of the range analysis and analysis of variance (ANOVA), the maximum biodiesel yield was found at a molar ratio of methanol to oil of 8:1, a reaction time of 70 min, a reaction temperature of 50 °C, and a catalyst concentration of 1 wt.%. The product and FAME yields of biodiesel under optimal conditions reached 95.8% and 98.4%, respectively. The properties of the optimized biodiesel, including density, kinematic viscosity, acid value, etc., were determined and compared with those produced from other oil feedstocks. The optimized biodiesel from camelina oil meets the relevant ASTM D6571 and EN 14214 biodiesel standards and can be used as a qualified fuel for diesel engines.     

Biodiesel production from mahua (Madhuca indica) oil having high free fatty acids

A technique to produce biodiesel from mahua oil (Madhuca indica) having high free fatty acids (19% FFA) has been developed. The high FFA level of mahua oil was reduced to less than 1% by a two-step pretreatment process. Each step was carried out with 0.30–0.35 v/v methanol-to-oil ratio in the presence of 1% v/v H₂SO₄ as an acid catalyst in 1-hour reaction at 60°C. After the reaction, the mixture was allowed to settle for an hour and methanol–water mixture that separated at the top was removed. The second step product at the bottom was transesterified using 0.25 v/v methanol and 0.7% w/v KOH as alkaline catalyst to produce biodiesel. The fuel properties of mahua biodiesel were found to be comparable to those of diesel and conforming to both the American and European standards.     

Eco-friendly biodiesel production from olive oil waste using solar energy

This study was carried out to produce biodiesel from olive oil waste by transesterification reaction. Several important reaction variables (the weight ratio of oil to methanol, the temperature, and reaction time) were evaluated to obtain a high quality of biodiesel fuel that meets authentic standards. Solar energy was applied for the transesterification reaction and electricity generated by photovoltaic panels was used to power a motor for mixing the reaction solution.     

Biodiesel production from degummed Jatropha curcas oil using constant-temperature ultrasonic water bath

This paper studied tri-basic potassium phosphate for transesterification process with degummed crude Jatropha curcas oil using constant-temperature, ultrasonic water bath generating low-intensity pulses with good energy distribution converting the maximum amount of biodiesel. Tri-basic potassium phosphate is suitable for J. curcas oil when the free fatty acid (FFA) content is less than 2%. The optimal reaction levels are catalyst 1.0 wt%, temperature of 50°C, and methanol-to-oil molar ratio of 12:1. The yield is 98% after 45 min, at 20 kHz frequency. The catalytic activity is found similar to potassium hydroxide and the catalyst solubility is only 4.27 ppm.     

Enzymatic transesterification of soybean oil with ethanol using lipases immobilized on highly crystalline PVA microspheres

Polyvinyl alcohol (PVA) microspheres with different degree of crystallinity were used as solid supports for Rhizomucor miehei lipase immobilization, and the enzyme-PVA complexes were used as biocatalysts for the transesterification of soybean oil to fatty acid ethyl esters (FAEE). The amounts of immobilized enzyme on the polymeric supports were similar for both the amorphous microspheres (PVA4) and the high crystalline microspheres (PVA25). However, the enzymatic activity of the immobilized enzymes was depended on the crystallinity degree of the PVA microspheres: enzymes immobilized on the PVA4 microspheres have shown low enzymatic activity (6.13 U mg⁻¹), in comparison with enzymes immobilized on the high crystalline PVA25 microspheres (149.15 U mg⁻¹). A synergistic effect was observed for the enzyme-PVA25 complex during the transesterification reaction of soybean oil to FAEE: transesterification reactions with free enzyme with the equivalent amount of enzyme that were immobilized onto the PVA25 microspheres (5.4 U) have yielded only 20% of FAEE, reactions with the pure highly crystalline microsphere PVA25 have not yielded FAEE, however reactions with the enzyme-PVA25 complexes have yielded 66.3% of FAEE. This synergistic effect of an immobilized enzyme on a polymeric support has not been observed before for transesterification reaction of triacylglycerides into FAEE. Based on ATR-FTIR, ²³Na- and ¹³C-NMR-MAS spectroscopic data and the interaction of the polymeric network intermolecular hydrogen bonds with the lipases residual amino acids a possible explanation for this synergistic effect is provided.     

Biodiesel production from waste sunflower oil by using Amberlyst 46 as a catalyst

In this study, simultaneous transesterification and esterification of high acid value sunflower oil to fatty acid methyl esters was studied using Amberlyst 46 as a heterogeneous catalyst. The influence of reaction conditions such as molar ratio of methanol/oil, reaction time, and reaction temperature was investigated. The highest fatty acids methyl esters yield of 75.8% was obtained in presence of 6 wt% oleic acid content under reaction conditions of 20 wt% Amberlyst 46 catalyst amount, 6/1 methanol/oil molar ratio, reaction temperature of 130°C, and reaction time of 10 h.