Enzymatic large‐scale production of biodiesel

Enzymatic production of biodiesel has been the subject of intensive research over the last 5‐10 years. When comparing the economy of enzymatic and chemical produced biodiesel important factors include yield, flexibility in feedstock, value of by‐products, recovery costs for alcohol as well as the cost of enzyme, enzyme lifetime, reaction time are important factors. By 2009 large‐scale enzymatic biodiesel was considered to be cost effective [1]. Since then development work has further improved the technology which is now ready for the market. We have developed a full enzymatic two‐step process for converting oils to biodiesel, and a pretreatment unit which can be installed in an existing chemical biodiesel plant to enable it to process high free fatty acid feedstocks.     

Enzymes in lipid production and modification – Current knowledge and future perspectives

Microbial enzymes are today used in a range of lipid processing from refining through to fat modification. The range of options is increasing following new research into enzyme types and functionalities. Within the near future, improved methods of enzymatic degumming will be developed as well as alternative enzymes for interesterification and synthesis of nutritionally interesting fats. An enzymatic route to Biod‐diesel is also a distinct possibility focussing on the waste oils and fats which are today difficult to process through chemical esterification.     

Production of Biodiesel Using Liquid Lipase Formulations

Looking back at the literature for enzymatic biodiesel, it is evident that the research has been focused on using immobilized lipase to enable re‐use of the enzyme due to price constraints on lipases used for catalyzing the transesterification process. The use of liquid formulations of lipase for biodiesel has recently been implemented in the industry. Technology for using liquid formulated lipases for enzymatic biodiesel production is new and, since enzyme prices have been reduced, it is now possible to simplify the process considerably and apply it for very low‐quality oils. In this paper, the use of liquid lipase formulations for enzymatic biodiesel will be described along with a general proposal for an industrial‐scale enzymatic biodiesel process with >95 % yield.     

The application of biotechnological methods for the synthesis of biodiesel

Synthesis of biodiesel is performed mainly by chemical catalysis, but can also be performed by enzymatic or microbial methods, and these might play an important role in future substitution of petroleum‐based diesel. To discover sustainable, economically attractive biotechnological processes for biodiesel synthesis, close cooperation between different disciplines is needed. Currently, lipases are the enzymes of choice for the synthesis of fatty acid esters (FAE) from fats and oils, yielding biodiesel with the methyl esters (FAME) as the most important product. More recently, the direct production of FAE using engineered whole cell microorganism has also been described (MicroDiesel). Current enzymatic processes are still hampered by the high costs of the biocatalyst, but significant progress has recently been made leading to the first industrial enzymatic biodiesel production. Enzymatic biodiesel production is mostly attractive because of the starting materials (waste frying oils, oils with high water content, etc.), for which conventional chemical interesterification can hardly be applied.     

Enzymatic transesterification for production of biodiesel using yeast lipases: An overview

Biodiesel has provided an eco-friendly solution to fuel crisis, as it is renewable, biodegradable and a non-toxic fuel that can be easily produced through enzymatic transesterification of vegetable oils and animal fats. Enzymatic production of biodiesel has many advantages over the conventional methods as high yields can be obtained at low reaction temperatures with easy recovery of glycerol. Microbial lipases are powerful biocatalysts for industrial applications including biodiesel production at lower costs due to its potential in hydrolyzing waste industrial materials. Among them, lipases from yeasts, Candida antarctica, Candida rugosa, Cryptococcus sp., Trichosporon asahii and Yarrowia lipolytica are known to catalyze such reactions. Moreover, stepwise addition of methanol in a three step, two step and single step reactions have been developed using yeast lipases to minimize the inhibitory effects of methanol. The latest trend in biodiesel production is the use of whole-cell as biocatalysts, since the process requires no downstream processing of the enzyme. Synthesis of value added products from the byproduct glycerol further reduces the production cost of biodiesel. This review aims at compiling the information on various yeast lipase catalyzed transesterification reactions for greener production of biodiesel.     

Versuchsplanerische Untersuchung der Umesterung von Sojaöl mit Ethanol im Mikroreaktor

Base‐catalyzed transesterification of fats and oils with primary alcohols in discontinuous operation is an established batch process for the biodiesel production. However, the application of microreaction technology and continuous flow process lead to an increase of process intensification. The ethanol/soy bean oil ratio at low flow rates as well as the reactor geometry have the most evident effects on the fatty acid ethyl ester yield of KOH‐catalyzed ethanolysis of soy bean oil in microreactors. The influence of the catalyst concentration is of a lower importance.     

Interesterification,chemical or enzymatic catalysis

The interesterification process is one of the oil modification processes the refiner can use to alter the physical properties of oils and fats, The reaction requires a catalyst to proceed. This can be a base or a lipase enzyme. In the currently accepted mechanism of the base‐catalysed interesterification reaction, two anionic intermediates are involved: the enolate anion and the glycerolate anion. The presence of the enolate anion explains why an equivalent amount of FAMEs are formed when sodium methanolate is added to oil and why FFAs are formed when the catalysts is inactivated with water. Based on this insight, process development can aim at avoiding these by‐products and thereby increase the cost advantage of the chemical process over the enzymatic process even further. The chemical process is also more flexible than the solely continuous enzymatic process, which latter requires extensive purification of the oil to be interesterified.     

Lipase-Catalyzed Production of Biodiesel by Hydrolysis of Waste Cooking Oil Followed by Esterification of Free Fatty Acids

Biodiesel is conventionally produced by alkaline‐catalyzed transesterification, which requires high‐purity oils. However, low‐quality oils can be used as feedstocks for the production of biodiesel by enzyme‐catalyzed reactions. The use of enzymes has several advantages, such as the absence of saponification side reactions, production of high‐purity glycerol co‐product, and low‐cost downstream processing. In this work, biodiesel was produced from lipase‐catalyzed hydrolysis of waste cooking oil (WCO) followed by esterification of the hydrolyzed WCO (HWCO). The hydrolysis of acylglycerols was carried out at 30 °C in salt‐free water (WCO/water ratio of 1:4, v/v) and the esterification of HWCO was carried out at 40 °C with ethanol in a solvent‐free medium (HWCO/ethanol molar ratio of 1:7). The hydrolysis and esterification steps were carried out using immobilized Thermomyces lanuginosus lipase (TLL/WCO ratio of 1:5.6, w/w) and immobilized Candida antarctica lipase B (10 wt%, CALB/HWCO) as biocatalysts, respectively. The hydrolysis of acylglycerols was almost complete after 12 h (ca. 94 %), and in the esterification step, the conversion was around 90 % after 6 h. The purified biodiesel had 91.8 wt% of fatty acid ethyl esters, 0.53 wt% of acylglycerols, 0.003 wt% of free glycerol, viscosity of 4.59 cP, and acid value of 10.88 mg KOH/g. Reuse hydrolysis and esterification assays showed that the immobilized enzymes could be recycled five times in 10‐h batches, under the conditions described above. TLL was greatly inactivated under the assay conditions, whereas CALB remained fully active. The results showed that WCO is a promising feedstock for use in the production of biodiesel.     

Optimal integration for biodiesel production using bioethanol

The production of biodiesel from algae is optimized using bioethanol following four different transesterification paths: alkali, enzymatic, and heterogeneous catalysts and supercritical conditions. The reactors are modeled using response surface methodology based on experimental results from the literature. These reactor models are implemented together with short‐cut methods for the other equipment (distillation columns, gravity separators, etc.) in order to recover the ethanol, separate the polar and nonpolar phases, and purify the glycerol and biodiesel produced to formulate the problem as a superstructure of alternatives. The aim is to simultaneously optimize and heat integrate the production of biodiesel using ethanol in terms of the reaction technology and the operating conditions. The optimal conditions in the reactors differ from the ones traditionally used because these results take the separation stages into account. In terms of the optimal process, the alkali catalyzed process is the most profitable, while the enzymatic one is also promising due to the lower consumption of energy and water, although it requires significant enzyme cost. © 2012 American Institute of Chemical Engineers AIChE J, 59: 834–844, 2013     

Catalytic chemistry of preparation of hydrocarbon fuels from vegetable oils and fats

The problem of preparing engine fuels from renewable feedstocks via the catalytic processing of inedible vegetable oils and fats is considered. Different types of inedible feedstocks are described, including algae, inedible plants, wood processing products, and waste fats and oils. Catalytic processes are considered for preparing the second generation biodiesel through the hydrodeoxygenation and deoxygenation of triglycerides and fatty acids, and of their derivatives. Brief information on catalysts for the deoxygenation of fatty acids is given. Special attention is given to analyzing the mechanism and kinetics of the deoxygenation reaction. Based on conducted kinetic and quantum-chemical investigations and using the literature data, a deoxygenation mechanism is proposed by the authors that explains the observed dependences of decarboxylation and decarbonylation contributions on the reaction conditions (the stearic acid, water, and catalyst concentrations, the hydrogen and CO pressures, and the temperature). Examples of the application of hydrocarbon biodiesel in transport are presented.     

Fast analysis of fats,oils and biodiesel by FT‐IR

Historically, the determination of chemical information or traits which describe the quality of fats and oils required analysis using wet chemistry, chromatography, and spectroscopic methods. Unfortunately, each of these methods is time‐consuming and requires a well equipped laboratory staffed with trained chemists. In contrast, Fourier Transform Infrared (FT‐IR) Spectroscopy is a fast analytical technique, but requires expertise for method development and operation. To overcome these difficulties, we have successfully developed a FT‐IR system which utilizes remote/local FT‐IR data collection, Internet communication to a central database, FT‐IR based chemometric analysis, and re‐communication of the analysis results to the remote unit. This Internet‐enabled quality trait analysis (QTA®) FT‐IR system has been thoroughly field tested. It has been found to be a rugged, fast, and accurate technique for the analysis of fats and oils and biodiesel. In addition, the Internet‐enabled QTA® FT‐IR system requires minimal skills for operation. Examples using the Internet‐enabled QTA® FT‐IR system for the analysis of fats and oils and biodiesel are described in this article.     

Oxidation stability of biodiesel fuel as prepared by supercritical methanol

A non-catalytic supercritical methanol method is an attractive process to convert various oils/fats efficiently into biodiesel. To evaluate oxidation stability of biodiesel, biodiesel produced by alkali-catalyzed method was exposed to supercritical methanol at several temperatures for 30 min. As a result, it was found that the tocopherol in biodiesel is not stable at a temperature higher than 300 °C. After the supercritical methanol treatment, hydroperoxides were greatly reduced for biodiesel with initially high in peroxide value, while the tocopherol slightly decreased in its content. As a result, the biodiesel prepared by the supercritical methanol method was enhanced for oxidation stability when compared with that prepared by alkali-catalyzed method from waste oil. Therefore, supercritical methanol method is useful especially for oils/fats having higher peroxide values.     

动植物油生产清洁燃料和低碳烯烃的替代加工工艺

Since the production cost of biodiesel is now the main hurdle limiting their applicability in some areas, catalytic cracking reactions represent an alternative route to utilization of vegetable oils and animal fats. Hence, catalytic transformation of oils and fats was carried out in a laboratory-scale two-stage riser fluid catalytic cracking （TSRFCC） unit in this work. The results show that oils and fats can be used as FCC feed singly or co-feeding with vacuum gas oil （VGO）, which can give high yield （by mass）of liquefied petroleum gas （LPG）, C2-C4 oletms, tor example 45% LPG, 47% C2-C4 olefins, and 77.6% total liquid yield produced with palm oil cracking. Co-feeding with VGO gives a high yield of LPG （39.1%） and propylene （18.1%）. And oxygen element content is very low （about 0.5%） in liquid products, hence, oxygen is removed in the form of H2O, CO and CO2. At the same time, high concentration of aromatics （C7-C9 aromatics predominantly） in the gasoline fraction is obtained after TSRFCC reaction of palm oil, as a result of large amount of hydrogen-transfer, cyclization and aromatization reactions, Additionally, most of properties of produced gasoline and diesel oil fuel meet the requirements of national standards, containing little sulfur. So TSRFCC technology is thought to be an alternative processing technology leading to production of clean fuels and light olefins.     

Biotechnological valorization of biodiesel derived glycerol waste through production of single cell oil and citric acid by Yarrowia lipolytica

Continuous energy crises and increasing demand for conventional fuels has resulted in the need for biofuels on a commercial scale. Transesterification of oils to yield biodiesel – one of the principal biofuels currently produced in large‐scale operations – is coupled with significant production of a glycerol‐rich water (so‐called “crude” or “raw” glycerol), as an important side‐product of the process. The increasing demand for biodiesel leads to abundant quantities of this glycerol‐rich material on the market. Therefore, glycerol valorization has much to offer in the cost reduction of biodiesel production. To this end, various chemical or biotechnological strategies have been developed to obtain added‐value products using crude glycerol as substrate. This review combines an account of our attempts to achieve a biotechnological valorization of raw glycerol with a review of appropriate literature.     

Application of a microassay method to study enzymatic hydrolysis of pretreated wheat straw

BACKGROUND: The conversion of lignocellulosic biomass to ethanol includes a disruptive pretreatment process followed by enzyme‐catalyzed hydrolysis of the cellulose and hemicellulose components to fermentable sugars. As the cost and hydrolytic efficiency of enzymes are major factors that restrict the commercialization of biomass conversion processes, significant efforts are made nowadays to improve the enzymatic mixtures and make the process cost‐effective. RESULTS: In this work, enzymatic microassays have been developed and validated to test new different enzymatic formulations on real lignocellulosic substrates. Homogeneous handsheets from steam pretreated wheat straw were elaborated to be used as substrate. The microassay was adapted to test both water‐insoluble solids and the whole slurry as substrates. Results in hydrolysis microassays were comparable with those obtained in standard flask assays using pretreated wheat straw. Moreover, using the enzymatic microassays, two novel preparations have been evaluated, demonstrating the ability of microassays to discriminate between different enzymatic mixtures. CONCLUSIONS: This enzymatic microassay represents a rapid method to test the performance of new selected cellulase enzymes on real pretreated lignocellulosic substrates. This microassay will enable evaluation of enzyme components separately, or optimized mixtures. Copyright © 2010 Society of Chemical Industry     

A review of conventional and renewable biodiesel production

The need for sustainable fuels has resulted in the production of renewables from a wide range of sources, in particular organic fats and oils. The use of biofuel is becoming more widespread as a result of environmental and economic considerations. Several efforts have been made to substitute fossil fuels with green fuels. Ester molecules extracted from processed animal fats and organic plant materials are considered alternatives for the use in modern engine technologies. Two different methods have been adopted for converting esters in vegetable oils/animal fats into compounds consistent with petroleum products, namely the transesterification and the hydro-processing of ester bonds for the production of biodiesel. This review paper primarily focuses on conventional and renewable biodiesel feedstocks, the catalyst used and reaction kinetics of the production process.     

Experimental Design Applied for Cost and Efficiency of Antioxidants in Biodiesel

Oxidation stability is a parameter of great importance for biodiesel quality control to both producers and subsequent consumers. To maintain the quality of biodiesel, currently the most effective and economical method is the addition of antioxidants that prevent or retard the biofuel oxidation reaction. In this study, efficiency and cost of synthetic antioxidants added to B100 biodiesel from soybean oil and pork fat were evaluated, using butylhydroxyanisole (BHA), butylhydroxytoluene (BHT) and tert-butylhydroquinone (TBHQ), in pure form or in mixtures, according to a simplex-centroid mixture experimental design. Results demonstrate an increased induction period (IP) in all trials when compared to the control sample, and TBHQ was the only antioxidant alone that met all the specification standards, while BHT and BHA alone met only the American standard specifications. The antioxidant mixture that presented the highest synergistic effect was that of TBHQ and BHA. Multi-response optimization indicated an optimum formulation containing 75 % TBHQ and 25 % BHA with an IP of 7.27 h at 110 °C and the antioxidant mixture cost of 31.31 USD, to be added for a ton of biodiesel. This simplex-centroid mixture experimental design shows an ability to be applied in the biodiesel, oils and fats industry to evaluate the oxidation stability and the occurrence of synergism between different mixtures of synthetic or natural antioxidants and their costs.     

Biodiesel production from high FFA rubber seed oil

Currently, most of the biodiesel is produced from the refined/edible type oils using methanol and an alkaline catalyst. However, large amount of non-edible type oils and fats are available. The difficulty with alkaline-esterification of these oils is that they often contain large amounts of free fatty acids (FFA). These free fatty acids quickly react with the alkaline catalyst to produce soaps that inhibit the separation of the ester and glycerin. A two-step transesterification process is developed to convert the high FFA oils to its mono-esters. The first step, acid catalyzed esterification reduces the FFA content of the oil to less than 2%. The second step, alkaline catalyzed transesterification process converts the products of the first step to its mono-esters and glycerol. The major factors affect the conversion efficiency of the process such as molar ratio, amount of catalyst, reaction temperature and reaction duration is analyzed. The two-step esterification procedure converts rubber seed oil to its methyl esters. The viscosity of biodiesel oil is nearer to that of diesel and the calorific value is about 14% less than that of diesel. The important properties of biodiesel such as specific gravity, flash point, cloud point and pour point are found out and compared with that of diesel. This study supports the production of biodiesel from unrefined rubber seed oil as a viable alternative to the diesel fuel.     

Bleaching of edible fats and oils

Almost all fats and oils are subjected to so‐called bleaching during processing. Originally bleaching was only used to reduce the colour. Today, however, the bleaching step is used mainly to remove or convert undesired by‐products to harmless ones from fats and oils. This will guarantee that such compounds do not interfere with the processing and that the requirements for human food are being met.     

Risk assessment for prion protein reduction under the conditions of the biodiesel production process

Due to the increased demand for biofuels, all different feedstocks from oils and fats have to be considered for biodiesel production. Animal fats have proved to be excellent sources for biodiesel due to their high cetane number and good stability. Large amounts of fat from so‐called high‐risk material, possibly contaminated with infectious prions, are available for biodiesel production. In this paper, the grade of destruction of prions during the biodiesel production process, including pre‐esterification with conc. sulfuric acid followed by KOH‐catalyzed transesterification, was studied. The starting material of the different production steps was spiked with purified and highly infectious prion rods, and the destruction of these prions was determined by gel electrophoresis (SDS‐PAGE) and Western blot. The pre‐esterification step led to a destruction factor of at least 100, the transesterification led to a factor of at least 250, and the distillation of the final biodiesel showed a destruction factor of at least 1000. During all experiments, no traces of prions could be detected after the different reaction steps. Based on these data, a complete and unequivocal risk assessment regarding the industrial process of biodiesel production was carried out, leading to a calculated overall risk of 5.8×10^?15 ID₅₀ units/person and year, which means that a hypothetical BSE contamination from biodiesel is more than 10⁹ times lower than the background risk.