Effect of room temperature ionic liquid structure on the enzymatic acylation of flavonoids

Enzymatic acylation reactions of flavonoids (rutin, esculin) with long chain fatty acids (palmitic, oleic acids) were carried out in 14 different ionic liquid media containing a range of cation and anion structures. Classification of RTILs according to flavonoid solubility (using COSMO-RS) was the basis for structural selection. Overall, anion selection had a far greater influence on lipase activity than choice of cationic moiety. RTILs containing TF₂N⁻, PF₆⁻ and BF₄⁻ anions were most successful as reaction media while RTILs containing anions with stronger solvating properties (i.e. H-bonding ability) resulted in decreased yields, likely due to increased interactions with the protein structure of the lipase. Biosynthesis of rutin proceeded much slower than of esculin. All-in-all, judicious selection of RTILs was central to achieving high yields (>98% after 6 days for TOMA·TF₂N) since a balance must be struck that maximized flavonoid solubility with minimum negative impact on lipase activity. The process also benefitted from an increased reaction temperature which may have helped to reduced mass transfer limitations.     

Studies on Guizotia abyssinica L. oil: Biodiesel synthesis and process optimization

Guizotia abyssinica seeds, a common bird feedstock, have been explored for the potential of biodiesel synthesis. The oil was extracted from the seeds by solvent extraction and composition of G. abyssinica oil was examined. The reaction parameters for biodiesel synthesis have been optimized. Temperature, oil: methanol ratio, catalyst type and catalyst concentration were found to have significant role on ester conversion. According to this study, the maximum yield of ester (98.7%) can be obtained with optimized sodium methoxide catalyst dosage (0.6%) at an operational temperature of 65 °C. Methyl ester of G. abyssinica oil was also studied for its oxidation stability and low temperature properties. Further, the synthesized product was blended in diesel at 5–20% ratios and evaluated for physico-chemical properties.     

Isolation and purification of lactoferrin and immunoglobulin G from bovine colostrum with serial cation-anion exchange chromatography

High-value dairy proteins such as lactoferrin (LF) and immunoglobulin (IgG) were separated from bovine colostrum. The whey was initially adjusted to pH 6.8 with 1 mol/L NaOH and then went through centrifugation, precipitation, and filtration to eliminate the fat and caseins in bovine colostrum. The treated whey was further ultra-filtrated to partially remove both other proteins and carbohydrates under 50 kD molecular weight. Then the ultra-filtrated whey was passed through cation and anion exchange columns in series. The LF and IgG were adsorbed on cation and anion exchanger, respectively, due to their different pI. Both the cation and anion exchange columns were washed with de-ionized water followed by successive elution with sodium chloride solutions of increasing molarities (0.27 and 0.85 mol/L; 17 and 51 mmol/L) in a stepwise manner, respectively. After desalted, the elution was freeze-dried. Finally, the LF and IgG with respective purities of 95.0% and 96.6% were obtained.     

Enhancing biodiesel production by immobilized whole cells by optimizing reaction conditions and adding glycerol and water

In this study, several methods were devised and evaluated to enhance biodiesel production by whole cells immobilized onto the polyurethane foam coated with activated carbon. Biodiesel conversion was increased to 76.4% with the increase in the number of polyurethane foam until it occupied 18.0 or 2.4% of reaction mixture based on apparent or actual volume of supports, respectively. Stepwise methanol addition to prevent methanol inhibition on the immobilized whole cells was optimized in terms of number of aliquot and feeding interval. When 4.5 molar ratio of methanol to soybean oil was divided into 4 equal aliquots and each aliquot was fed to the reaction mixture every 24 h, the highest final biodiesel conversion of 82.4% was achieved. Chemical treatment of the immobilized cells with 0.1% of chloroform for 2 h enhanced biodiesel conversion to 90.5%. The initial addition of 5% glycerol in the fresh reaction mixture increased biodiesel conversion to 90.3% while the removal of glycerol during biodiesel production barely increased biodiesel conversion. The biodiesel conversion was increased with the increase of initial water content in the fresh reaction mixture and the highest value was 92.7% at 3.0% of water content, but decreased thereafter. The effects of co-addition of glycerol and water on biodiesel production were also investigated, and the co-addition of 3.125% of glycerol and 1.875% of water relative to soybean oil substantially increased biodiesel conversion to 95.0%. By these optimization of reaction conditions and co-adding glycerol and water, initial biodiesel production rate and final biodiesel conversion were remarkably enhanced by 26.8 and 24.1%, respectively.     

Statistical analysis and optimization of biodiesel production from waste coffee grounds by a two-step process

Biodiesel was produced using waste coffee grounds (WCGs) via a two-step process comprising lipid extraction and subsequent transesterification steps. Each step was statistically analyzed, and optimum conditions for each step were suggested. WCGs were found to have 16.4% lipid content with 1.9% free fatty acid (FFA) content. The liquid-solid ratio (LSR) significantly influenced lipid extraction from WCGs, while extraction time and temperature did not; 92.7% of lipid extraction efficiency was achieved at 13.7 mL-hexane/g-WCGs, 30 min of extraction time, and 25°C. Owing to the relatively low FFA content, an alkaline catalyst (NaOH) reaction was used that requires less amount of catalyst, methanol, and shorter reaction time compared to an acid catalyst reaction. Reaction time and temperature were the major factors affecting biodiesel conversion, and 94.0% of biodiesel conversion was obtained at optimum conditions for transesterification: 0.5% catalyst, 1.5 mL-methanol/g-lipid, 45°C, and 9 h of reaction time. With the use of statistical analysis tools, high lipid extraction efficiency and biodiesel conversion were achieved at relatively mild conditions, which would reduce biodiesel production cost substantially.     

Biodiesel production by simultaneous extraction and conversion of total lipids from microalgae, cyanobacteria, and wild mixed-cultures

Microalgae have been identified as a potential biodiesel feedstock due to their high lipid productivity and potential for cultivation on marginal land. One of the challenges in utilizing microalgae to make biodiesel is the complexities of extracting the lipids using organic solvents followed by transesterification of the extracts to biodiesel. In the present work, reaction conditions were optimized that allow a single step extraction and conversion to biodiesel in high yield from microalgae. From the optimized conditions, it is demonstrated that quantitative conversion of triglycerides from several different microalgae and cyanobacteria could be achieved, including from mixed microbial biomass collected from a municipal wastewater lagoon. Evidence is presented that for some samples, significantly more biodiesel can be produced than would be expected from available triglycerides, indicating conversion of fatty acids contained in other molecules (e.g., phospholipids) using this approach. The effectiveness of the approach on wet algae is also reported.     

Penicillium expansum lipase-catalyzed production of biodiesel in ionic liquids

Penicillium expansum lipase (PEL) was used to catalyze biodiesel production from corn oil in [BMIm][PF₆]¹ (an ionic liquid, IL) and tert-butanol. Both systems were optimized in terms of MeOH/oil molar ratio, reaction temperature, enzyme loading, solvent volume, and water content. The high conversion obtained in the IL (86%) as compared to that in tert-butanol (52%) demonstrates that the IL is a superior solvent for PEL-catalyzed biodiesel production. Poor yields were obtained in a series of hydrophilic ILs. Addition of salt hydrates affected biodiesel production predominantly through the specific ion (Hofmeister) effect. The impact of methanol on both activity and stability of PEL in the IL and in hexane was investigated, in comparison to the results obtained by two commonly used lipases, Novozym 435 and Lipozyme TLIM. The results substantiate that while different lipases show different resistance to methanol in different reaction systems, PEL is tolerant to methanol in both systems.     

Structures of ionic liquids dictate the conversion and selectivity of enzymatic glycerolysis: theoretical characterization by COSMO-RS

Lipase-catalyzed glycerolysis of triolein has been examined using a group of tetraammonium-based ionic liquids (ILs) as media, specifically with functional groups in cation part. The results demonstrated that the reaction evolution and profile specificity of respective IL system could be quantitatively associated with the structural characteristics of the IL by means of quantum chemical and COSMO-RS calculation. Misfit interaction, Van der Waals interaction and chemical potential, etc. derived from COSMO-RS calculation are shown to be effective measures to delineate multiple interactions of ILs and then can be used to understand the effects of ILs on reactions. The hydrophobic substituents in the cation are found to contribute to the increase of triolein solubility and enhancement of initial reaction rate; while strong polar anion and polyethoxyl and free hydroxyl groups in the cation part dictate improved product selectivity through reducing activity coefficients of monoglycerides. Integration of these structures into the same molecule constitutes a promising group of ILs that could produce over 90% monoglyceride with almost 100% triglyceride conversion, as well as bulky productivity, of particular potential for industrial applications. Overall, this work has presented a first attempt to characterize the IL structure-dependency of reaction specificity by associating structural variations of ILs with thermodynamic property changes of resided compounds and subsequent effects on reaction specificity. This might be of general value to help to understand the multiple solvation interaction among IL reaction systems at molecular level and promote the application of IL-mediated reactions to practical interests.     

Biodiesel production in packed-bed reactors using lipase-nanoparticle biocomposite

The development of appropriate reactors is crucial for the production of biodiesel. In this study, a packed-bed reactor system using lipase-Fe(3)O(4) nanoparticle biocomposite catalyst was successfully developed for biodiesel production based on soybean oil methanolysis. Emulsification before methanolysis improved the reaction rate. The lipase-nanoparticle biocomposite showed high activity and stability in the single-packed-bed reactor at an optimal flow rate (0.25 mL min(-1)). After 240 h of reaction, the conversion rate was sustained as high as 45%. The conversion rate and stability achieved using the four-packed-bed reactor were much higher than those achieved using the single-packed-bed reactor. The conversion of biodiesel was maintained at a high rate of over 88% for 192 h, and it only slightly declined to approximately 75% after 240 h of reaction. The packed-bed reactor system, therefore, has a great potential for achieving the design and operation of enzymatic biodiesel production on the industrial scale.     

Optimization of a Novel Two-step Process Comprising Re-esterification and Transesterification in a Single Reactor for Biodiesel Production Using Waste Cooking Oil

Waste cooking oil (WCO) has attracted attention as a non-edible feedstock for biodiesel. Although an alkali catalyst has several advantages over an acid catalyst in biodiesel production, biodiesel conversion from WCO is only 5.2% when using an alkali catalyst (NaOH), owing to its high free fatty acid (FFA) content of 4.2%. In this study, a novel two-step process in a single reactor, comprised of re-esterification of the FFAs with crude glycerol, using a Tin (II) chloride (SnCl₂) catalyst, and subsequent transesterification with methanol, using an alkali catalyst, was adopted, and each step was optimized. This study revealed that the FFA content after re-esterification should be approximately 1.5%, not only to save glycerol and the catalyst involved in the re-esterification, but also to achieve high biodiesel conversion during the transesterification. An alkaline catalyst was successfully used to produce biodiesel in the second step, and a 92.8% conversion to biodiesel was achieved under the optimized conditions (0.6% catalyst relative to WCO, 0.2mL-methanol/WCO, 70ºC, 3 h). Overall, this novel two-step process achieved highly enhanced biodiesel conversion (4.0% to 92.8%) with significantly reduced reaction time (12 h to 4 h) and methanol requirements (15 mL/g-WCO to 0.2 mL/g-WCO).     

Biodiesel production from triolein and short chain alcohols through biocatalysis

Oleic acid alkyl esters (biodiesel) were synthesised by biocatalysis in solvent-free conditions. Different commercial immobilised lipases, namely Candida antarctica B, Rizhomucor miehei, and Pseudomonas cepacia, were tested towards the reaction between triolein and butanol to produce butyl oleate. Pseudomonas cepacia lipase resulted to be the most active enzyme reaching 100% of conversion after 6h. Different operative conditions such as reaction temperature, water activity, and reagent stoichiometric ratio were investigated and optimised. These conditions were then used to investigate the effect of linear and branched short chain alcohols. Methanol and 2-butanol were the worst alcohols: the former, probably, due to its low miscibility with the oil and the latter because secondary alcohols usually are less reactive than primary alcohols. Conversely, linear and branched primary alcohols with short alkyl chains (C(2)--C(4)) showed high reaction rate and conversion. A mixture of linear and branched short chain alcohols that mimics the residual of ethanol distillation (fusel oil) was successfully used for oleic acid ester synthesis. These compounds are important in biodiesel mixtures since they improve low temperature properties.     

Fractionation and lipase-catalyzed conversion of microalgal lipids to biodiesel

This study was conducted to evaluate the lipid fractionation and purification procedures of lipase-catalyzed conversion of neutral lipids to microalgal biodiesel. Microalgae lipids were efficiently recovered and purified by a combined extraction method and crude lipid extracts were separated into neutral lipids, glycolipids, and phospholipids by solid-phase extraction. The high purity of the neutral lipids fraction was confirmed by its low concentration of phosphorous (< 2.0 ppm). Transesterification was catalyzed by immobilized Candida antarctica lipase for 72 h with stepwise addition of methanol. The reaction displayed Michaelis–Menten kinetics and produced high yields of microalgal biodiesel (91.2% in the case of Dunaliella salina) with a high content of unsaturated fatty acids (81.5%). Neutral lipids were converted to biodiesel by three-step transesterification, while the removal of polar lipids maintained the activity of the immobilized lipase by reducing both reaction mixture viscosity and contamination risk.     

Asymmetric alkene epoxidation by Mn(III)salen catalyst in ionic liquids

The enantioselective epoxidation of 6-cyano-2,2-dimethylchromene (Chrom) catalysed by the Jacobsen catalyst, using sodium hypochlorite (NaOCl) as oxygen source, at room temperature, was performed in a series of 1,3-dialkylimidazolium and tetra-alkyl-dimethylguanidium based ionic liquids. All the room temperature ionic liquids (RTILs) could be used as reaction media for the enantioselective epoxidation of the alkene giving, generally, moderate to good epoxide yields and enantiomeric excesses (ee%).For the series of ionic liquids derived from the 1,3-dialkylimidazolium cation, it was observed some relationship between the RTILs physical properties and the catalytic reaction parameters, exemplified by linear correlations between (i) the ee% and the α Kamlet-Taft parameter (hydrogen bond acidity of the solvent) for CH₂Cl₂ and [C₄m_nim][BF₄] ionic liquids (n = 1 or 2), and (ii) the ee% and the β Kamlet-Taft parameter (hydrogen bond basicity of the solvent) for CH₂Cl₂ and [C₄mim][X] ionic liquids (X = PF₆, NTf₂ or BF₄).All the RTILs could be reused in further catalytic cycles, with the exception of [C₈mim][PF₆]. The reutilisation of the Jacobsen catalyst for four times generally led to a decrease in the epoxide yield and to a slight decrease in the enantioselectivity. The recycling of the catalyst could be improved by imparting an ionic character to the complex through abstraction of the axially coordinated chloride anion (Cat 2). Other oxygen sources, such as iodosylbenzene, hydrogen peroxide and urea-hydrogen peroxide adduct, were also tested coupled with Jacobsen catalyst, but the best results were achieved with NaOCl.     

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.     

Process for biodiesel production from Cryptococcus curvatus

The objective of the current report is process optimization for economical production of lipids by the well known oleaginous yeast Cryptococcus curvatus and conversion of the lipids to biodiesel. A high cell density fed-batch cultivation on low cost substrate viz. crude glycerol resulted in a dry biomass and oil yield of up to 69 g/L and 48% (w/w), respectively. The process was scaled up easily to 26 L. The oil extraction process was also optimized using environmentally safe solvents. The oil profile indicated a high oleic acid content followed by palmitic acid, stearic acid and linoleic acid. The oil was trans-esterified to biodiesel and thoroughly characterized. This is the first end to end report on production of biodiesel from the C. curvatus oil.     

Microwave assisted alkali-catalyzed transesterification of Pongamia pinnata seed oil for biodiesel production

In this study, microwave assisted transesterification of Pongamia pinnata seed oil was carried out for the production of biodiesel. The experiments were carried out using methanol and two alkali catalysts i.e., sodium hydroxide (NaOH) and potassium hydroxide (KOH). The experiments were carried out at 6:1 alcohol/oil molar ratio and 60 °C reaction temperature. The effect of catalyst concentration and reaction time on the yield and quality of biodiesel was studied. The result of the study suggested that 0.5% sodium hydroxide and 1.0% potassium hydroxide catalyst concentration were optimum for biodiesel production from P. pinnata oil under microwave heating. There was a significant reduction in reaction time for microwave induced transesterification as compared to conventional heating.     

Production of bioethanol and biodiesel using instant noodle waste

Instant noodle manufacturing waste was used as feedstock to convert it into two products, bioethanol and biodiesel. The raw material was pretreated to separate it into two potential feedstocks, starch residues and palm oil, for conversion to bioethanol and biodiesel, respectively. For the production of bioethanol, starch residues were converted into glucose by α-amylase and glucoamylase. To investigate the saccharification process of the pretreated starch residues, the optimal pretreatment conditions were determined. The bioethanol conversion reached 98.5 % of the theoretical maximum by Saccharomyces cerevisiae K35 fermentation after saccharification under optimized pretreatment conditions. Moreover, palm oil, isolated from the instant noodle waste, was converted into valuable biodiesel by use of immobilized lipase (Novozym 435). The effects of four categories of alcohol, oil-to-methanol ratio, reaction time, lipase concentration and water content on the conversion process were investigated. The maximum biodiesel conversion was 95.4 %.     

Influence of vegetable oils fatty acid composition on reaction temperature and glycerides conversion to biodiesel during transesterification

Presence of unreacted glycerides in biodiesel may reduce drastically its quality. This is why conversion of raw material in biodiesel through transesterification needs to readjust reaction parameter values to complete. In the present work, monitoring of glycerides transformation in biodiesel during the transesterification of vegetable oils was carried out. To check the influence of the chemical composition on glycerides conversion, selected vegetable oils covered a wide range of fatty acid composition. Reactions were carried out under alkali-transesterification in the presence of methanol. In addition, a multiple regression model was proposed. Results showed that kinetics depends on chemical and physical properties of the oils. It was found that the optimal reaction temperature depends on both length and unsaturation degree of vegetable oils fatty acid chains. Vegetable oils with higher degree of unsaturation exhibit faster monoglycerides conversion to biodiesel. It can be concluded that fatty acid composition influences reaction parameters and glycerides conversion, hence biodiesel yield and economic viability.     

Optimization of microwave-assisted transesterification of dry algal biomass using response surface methodology

The effect of microwave irradiation on the simultaneous extraction and transesterification (in situ transesterification) of dry algal biomass to biodiesel was investigated. A high degree of oil/lipid extraction from dry algal biomass and an efficient conversion of the oils/lipids to biodiesel were demonstrated in a set of well-designed experimental runs. A response surface methodology (RSM) was used to analyze the influence of the process variables (dry algae to methanol (wt/vol) ratio, catalyst concentration, and reaction time) on the fatty acid methyl ester conversion. Based on the experimental results and RSM analysis, the optimal conditions for this process were determined as: dry algae to methanol (wt/vol) ratio of around 1:12, catalyst concentration about 2 wt.%, and reaction time of 4 min. The algal biodiesel samples were analyzed with GC-MS and thin layer chromatography (TLC) methods. Transmission electron microscopy (TEM) images of the algal biomass samples before and after the extraction/transesterification reaction are also presented.     

Esterification of acidified oil with methanol by SPES/PES catalytic membrane

A sulfonated polyethersulfone (SPES)/polyethersulfone (PES) blend catalytic membrane was prepared and used as a heterogeneous catalyst in the esterification of the acidified oil (acid value 153 mg KOH/g) with methanol for producing biodiesel. The results showed that the free fatty acids conversion reached 97.6% using SPES/PES catalytic membrane under the optimal esterification conditions. Meanwhile, the SPES/PES membrane with 20.3% degree of sulfonation showed a good catalytic stability. A pseudo-homogeneous kinetic model was established. The results indicated that the reaction rate constant increased with increasing methanol/acidified oil molar ratio, the loading of catalytic membrane and reaction temperature. The reaction order was 2 and the activation energy decreased from 74.65 to 21.07 kJ/mol with increasing catalytic membrane loading from 0 to 0.135 meq/g(oil). It implies that the esterification is not diffusively controlled but kinetically controlled. The predicted results were in good agreement with the experimental data.