Showing papers on "Lipase published in 2010"
TL;DR: The current status of biodiesel production with immobilized lipase is reviewed, including various lipases, immobilization methods, various feedstocks, lipase inactivation caused by short chain alcohols and large scale industrialization.
618 citations
TL;DR: The main uses of the lipase from Thermomyces laguginosus will be revised, with special emphasis in the modification of fats and oils, production of biodiesel, resolution of racemic mixtures, enantioselective hydrolysis of prochiral esters and regioselectives process involving sugar preparations.
Abstract: The lipase from Thermomyces laguginosus (formerly Humicola laguginosa) (TLL) is a basophilic and noticeably thermostable enzyme, commercially available in both soluble and immobilized form. Although initially oriented toward the food industry, the enzyme has found applications in many different industrial areas, from biodiesel production to fine chemicals (mainly in enantio and regioselective or specific processes). This review intends to show some of the most relevant aspects of the use of this interesting enzyme. After checking the enzyme features, some of the most efficient methods of TLL immobilization will be commented. Finally, the main uses of the enzyme will be revised, with special emphasis in the modification of fats and oils, production of biodiesel, resolution of racemic mixtures, enantioselective hydrolysis of prochiral esters and regioselective process involving sugar preparations. In many instances, TLL has been compared to other lipases, the advantages or disadvantages of the enzyme will be discussed.
510 citations
TL;DR: In general, this enzyme seems very adequate for esterification reactions due to its high stability in anhydrous media and good esterization activity.
Abstract: The lipase from Rhizomucor miehei (formerly Mucor miehei) (RML) is a commercially available enzyme in both soluble and immobilized form with very high activity and good stability under diverse conditions (anhydrous organic solvents, supercritical fluids, etc.). Although this lipase was initially produced to be used in food industry, in this review we will focus our attention on the application of this enzyme in organic chemistry, from biodiesel production to fine chemicals (mainly in enantio or regioselective or specific processes). After showing the enzyme features, some of the most efficient methods of RML immobilization will be commented (entrapping on reverse micelles, preparation of cross-linked RML aggregates or immobilization on pre-existing solids). Finally, the main uses of the enzyme in organic chemistry will be revised. The use of RML in the production of biodiesel will be analyzed, and compared to the performance of other lipases. The synthesis of esters of carboxylic acids as flavors is other example where RML has been successfully employed. Taking advantage of the wide specificity of the enzyme, mainly a high enantiospecificity, many examples of the use of RML in the resolution of racemic mixtures of chiral carboxylic acids, alcohols or esters will be presented. Special mention requires the use of the regioselectivity of RML, mainly the chemistry of sugars. Finally, more unusual uses of RML will be presented (anomalous substrates, novel uses, etc.). In general, this enzyme seems very adequate for esterification reactions due to its high stability in anhydrous media and good esterification activity.
264 citations
TL;DR: A comprehensive review of the literature on the subject of biodiesel production was carried out in this paper, where the most popular microbes used for their lipases have been filamentous fungi and recombinant bacteria.
Abstract: Problem Statement: The research on the production of biodiesel has increased significantly in recent years because of the need for an alternative fuel which endows with biodegradability, low toxicity and renewability. Plant oils, animal fats, microalgal oils and waste products such as animal rendering, fish processing waste and cooking oils have been employed as feedstocks for biodiesel production. In order to design an economically and environmentally sustainable biodiesel production process, a proper understanding of the factors affecting the process and their relative importance is necessary. Approach: A comprehensive review of the literature on the subject of biodiesel production was carried out. Traditionally biodiesel has been produced using either acid or base catalysts. The multi-step purification of end products, wastewater treatment and energy demand of the conventional process has lead to search for alternative option for production of biodiesel. The use the enzyme lipase as a biocatalyst for the transesterification reaction step in biodiesel production has been extensively investigated. Lipase is produced by all living organisms and can be used intracellularly or extracellularly. Conclusion: To date, the most popular microbes used for their lipases have been filamentous fungi and recombinant bacteria. A summary of lipases used in transesterification and their optimum operating conditions is provided. In addition to the choice of lipase employed, factors which make the transesterification process feasible and ready for commercialization are: enzyme modification, the selection of feedstock and alcohol, use of common solvents, pretreatment of the lipase, alcohol to oil molar ratio, water activity/content and reaction temperature. Optimization of these parameters is necessary in order to reduce the cost of biodiesel production. Use of no/low cost waste materials as feedstocks will have double environmental benefits by reducing the environmental pollution potential of the wastes and producing an environmentally friendly fuel.
246 citations
TL;DR: The enzyme has been the first lipase whose structure was reported and its interfacial activation is very well known, making this enzyme a good template for modeling studies, and seems to be advantageous when compared to other lipases mainly in esterificaton reactions in anhydrous media.
Abstract: The lipase from Rhizomucor miehei (RML), formerly Mucor miehei, is a commercially available enzyme in both soluble and immobilized forms with very high activity and good stability under diverse experimental conditions (anhydrous organic solvents, supercritical fluids, etc.). The uses of the enzyme were initially oriented towards food industry, that way the enzyme has found a broad application in this area. This review intends to show the enzyme features and some of the most relevant aspects of the use of this interesting enzyme in oils and fats modification. The enzyme has been the first lipase whose structure was reported and its interfacial activation is very well known, making this enzyme a good template for modeling studies. Finally, the main uses of the enzyme in fatty acids, oils and fats modification will be revised (hydrolysis of glycerides, transesterification, esterification, acidolysis and interesterification). This lipase seems to be advantageous when compared to other lipases mainly in esterificaton reactions in anhydrous media, while other lipases can be preferred in transesterification reactions.
238 citations
TL;DR: There are strong epidemiologic connections between plasma triglycerides and atherosclerosis, and to what extent this goes back to derangements of the lipoprotein lipase (LPL) system is considered.
Abstract: Lipoprotein lipase is at the center of lipoprotein metabolism, unloading triglycerides for metabolic use in extrahepatic tissues, regulated in concert with energy metabolism by insulin and other factors, and converting the input of large triglyceride-rich lipoproteins to smaller, potentially atherogenic remnants that are either remodeled into LDL and HDL, or quickly cleared. In addition, lipoprotein lipase has turned out to be a multifunctional protein that also acts as a ligand for binding of lipoproteins to proteoglycans and receptors. Hepatic lipase has evolved more recently, and its main role appears to be in the remodeling processes.
211 citations
TL;DR: Pectin extracted from the apple pomace was evaluated for the in vitro inhibition of pancreatic lipase (steapsin) as discussed by the authors, and the effect of the extraction process on the structure of the extracted pectin was evaluated by the physico-chemical parameters and different techniques such as XRD, 13 C NMR, FTIR or Raman spectroscopy.
189 citations
01 Jan 2010
TL;DR: In vitro polyester synthesis catalyzed by a hydrolysis enzyme of lipase is described, most of which has been developed for these two decades, and the polymerization characteristics are of green nature currently demanded for sustainable society, and hence, desirable for conducting ‘green polymer chemistry’.
Abstract: This article is a short comprehensive review describing in vitro polyester synthesis catalyzed by a hydrolysis enzyme of lipase, most of which has been developed for these two decades. Polyesters are prepared by repeated ester bond-formation reactions; they include two major modes, ring-opening polymerization (ROP) of cyclic monomers such as cyclic esters (lactones) and condensation polymerization via the reaction between a carboxylic acid or its ester group and an alcohol group. Polyester synthesis is, therefore, a reaction in reverse way of in vivo lipase catalysis of ester bond-cleavage with hydrolysis. The lipase-catalyzed polymerizations show very high chemo-, regio-, and enantio-selectivities and involve various advantageous characteristics. Lipase is robust and compatible with other chemical catalysts, which allows novel chemo-enzymatic processes. New syntheses of a variety of functional polyesters and a plausible reaction mechanism of lipase catalysis are mentioned. The polymerization characteristics are of green nature currently demanded for sustainable society, and hence, desirable for conducting ‘green polymer chemistry’.
151 citations
TL;DR: In this article, the binding of lipase to magnetic particles was confirmed by enzyme assays, transmission electron microscopy (TEM) and Fourier transform infrared (FT-IR) spectra.
Abstract: Lipase was covalently immobilized onto magnetic Fe3O4 nano-particles by using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDAC) as an activating agent, and the bound lipase was used to catalyze the transesterification of vegetable oils with methanol to produce fatty acid methyl esters. The binding of lipase to magnetic particles was confirmed by enzyme assays, transmission electron microscopy (TEM) and Fourier transform infrared (FT-IR) spectra. It was determined that the immobilized lipase exhibited better resistance to temperature and pH inactivation in comparison to free lipase. Using the immobilized lipase, the major parameters affecting the transesterification reaction, such as the alcohol/oil molar ratio, enzyme loading and free fatty acid present in reactants were investigated to obtain the optimum reaction condition. The conversion of soybean oil to methyl esters reached over 90% in the three-step transesterification when 40% immobilized lipase was used. Moreover, the lipase catalyst could be used for 3 times without significant decrease of the activity.
150 citations
TL;DR: In this article, the effects of pore sizes on lipase distribution, relative activity, kinetic behavior, thermal stability, storage stability, and reusability were investigated in detail.
Abstract: Polystyrene (PST) microspheres are commonly chosen as immobilization carriers due to their unique advantages such as ideal mechanical strength, adjustable particle size, and favorable chemical stability. However, there were few reports on immobilization by using microspheres with large pore sizes (>100 nm). We have successfully prepared the PST microspheres with macropores and gigapores by a novel method. In this study, giga-/macro-/meso-porous PST microspheres (314 nm, 104 nm, and 14.7 nm in pore sizes) were employed to immobilize lipase (from Burkholderia cepacia) by strong hydrophobic interactions, and the effects of pore sizes on lipase distribution, relative activity, kinetic behavior, thermal stability, storage stability, and reusability were also investigated in detail. According to laser scanning confocal microscope (LSCM) observation, lipase penetrated into the center of those giga-/macro-porous microspheres. With regard to the mesoporous microspheres, lipase only adsorbed to the external shell. The relative activities of immobilized lipase were 146%, 126%, and 50.9% for giga-/macro-/meso-porous PST microspheres, respectively. Comparing with the kinetic constants of free lipase (0.441 mM), the K(m) value for mesoporous PST-lipase (0.532 mM) was higher, whereas for giga-/macro-porous PST-lipase (0.402 mM and 0.411 mM), the K(m) values were comparatively lower suggesting the accessibility of substrate to the enzyme active sites was unlimited. The thermal stability, storage stability, and reusability were all improved significantly with the increase of pore sizes. In stimulant system, even after 100 times of recycling, the activity of lipase immobilized on gigaporous and macroporous PST microspheres remained nearly 100% and 93%, respectively, while that of lipase-PST with 14.7 nm pores could only keep 64.1%. In real system, 73.5% and 68.8% activity of gigaporous PST-lipase and macroporous PST-lipase, respectively, were retained after being used 30 times, whereas only 49.4% activity was remained for mesoporous PST-lipase. Therefore, compared with other microspheres, the gigaporous PST microsphere exhibited obvious advantages as a potential enzyme support in industry. (C) 2010 Elsevier B.V. All rights reserved.
149 citations
Patent•
14 Dec 2010
TL;DR: In this article, the authors present compositions and methods relating to a lipase cloned from Thermobifida fusca, polynucleotides encoding the lipase, and methods of use thereof.
Abstract: The present compositions and methods relate to a lipase cloned from Thermobifida fusca, polynucleotides encoding the lipase, and methods of use thereof. The compositions and methods have particular application in detergent cleaning compositions and methods.
TL;DR: The present study suggests that Morus bombycis root extract might be of therapeutic interest with respect to the treatment of obesity.
TL;DR: Chitosan and glutaraldehyde were used as the "bridge unit" and "cross-linking agent", respectively, which were supposed to realize the immobilization of lipase molecules adsorbed on the surface of pores into a mesh-like layer, and the experimental results showed that the activities of the immobilized CRL were much higher than that of free lipase.
TL;DR: The results suggest that the lipid‐lowering effects of the 46‐kDa chitosan may be mediated by increases in faecal fat and/or bile acid excretion resulting from the binding of bile acids, and by a decrease in the absorption of dietary lipids from the small intestine as a result of the inhibition of pancreatic lipase activity.
Abstract: Three low molecular weight chitosans (molecular weight: 21, 46 and 130 kDa) obtained by enzymatic hydrolysis of a high molecular weight chitosan (average molecular weight: 650 kDa) had low viscosity and were water-soluble. The effects of these water-soluble chitosans on pancreatic lipase (in-vitro) and the elevation of plasma triacylglycerol concentration after the oral lipid tolerance test were examined in mice. The water-soluble 46-kDa chitosan was the most effective at inhibiting pancreatic lipase activity (in-vitro) and plasma triacylglycerol elevation after the oral lipid tolerance test. Based on this result, the effects of the 46-kDa chitosan on increases in bodyweight, various white adipose tissue weights, and plasma and liver lipids were examined in mice fed a high-fat diet for 20 weeks. Water-soluble 46-kDa chitosan (300 mg kg - 1 , twice daily) prevented increases in bodyweight, various white adipose tissue weights and liver lipids (cholesterol and triacylglycerol) in mice fed a high-fat diet, and further increased the faecal bile acid and fat. The results suggest that the lipid-lowering effects of the 46-kDa chitosan may be mediated by increases in faecal fat and/or bile acid excretion resulting from the binding of bile acids, and by a decrease in the absorption of dietary lipids (triacylglycerol and cholesterol) from the small intestine as a result of the inhibition of pancreatic lipase activity. Water-soluble 46-kDa chitosan (100 and 300 mg kg - 1 , twice daily) did not cause liver damage with the elevation of glutamic oxaloacetic transaminase and glutamic pyruvic transaminase, or kidney damage with the elevation of blood nitrogen urea. It was concluded that water-soluble 46-kDa chitosan is a safe functional food.
Patent•
16 Dec 2010
TL;DR: In this article, the authors present compositions and methods relating to a lipase cloned from Bacillus subtilis, polynucleotides encoding the lipase, and methods of use thereof.
Abstract: The present compositions and methods relate to a lipase cloned from Bacillus subtilis, polynucleotides encoding the lipase, and methods of use thereof. The compositions and methods have particular application in detergent cleaning compositions and methods.
Book Chapter•
01 Jan 2010
TL;DR: This chapter presents an overview of Yarrowia lipolytica features and its major biotechnological applications.
Abstract: Yarrowia lipolytica is one of the most extensively studied ‘‘non-conventional’’ yeasts, being a strictly aerobic microorganism capable of producing important metabolites and having an intense secretory activity, which justifies efforts to use it in industry (as a biocatalyst), in molecular biology and in genetics studies. Moreover, Y. lipolytica has been considered an adequate model for dimorphism studies in yeasts. Yarrowia lipolytica presents the ability to grow on Olive Mill Wastewater (OMW) as well as to degradate organic compounds, including aliphatic and aromatic hydrocarbons, often accompanied by biosurfactants production. One of the most important products secreted by this microorganism is lipase which can be exploited for several applications in the detergent, food, pharmaceutical, and environmental industries. In addition, Y. lipolytica is able to produce citric acid and aroma from a variety of carbon sources, including sugars, alkanes, plant oils, starch hydrolysates, ethanol, and glycerol. Thus, this chapter presents an overview of Yarrowia lipolytica features and its major biotechnological applications.
TL;DR: The EH28 lipase was used for synthesis of the flavor ester ethyl caprylate in organic solvents, thus providing a concept of application of Acinetobacter sp.
TL;DR: Structural and biochemical characterization have revealed that the hydrophobic microenvironment created by the organo-modified clays induces minor changes on the secondary structure of the enzyme, resulting in enhanced catalytic behaviour in hydrophilic media.
Patent•
14 Dec 2010
TL;DR: In this article, the authors present compositions and methods relating to a lipase cloned from Geobacillus stearothermophilus, polynucleotides encoding the lipase, and methods of use thereof.
Abstract: The present compositions and methods relate to a lipase cloned from Geobacillus stearothermophilus, polynucleotides encoding the lipase, and methods of use thereof. The compositions and methods have particular application in detergent cleaning compositions and methods.
TL;DR: Thermobifida alba strain AHK119 was shown to possess the ability to significantly degrade aliphatic-aromatic copolyester film as well as decreasing the polymer particle sizes when grown at 50°C on LB medium supplemented with polymer particles, yielding terephthalic acid.
Abstract: More than 100 bacterial strains were isolated from composted polyester films and categorized into two groups, Actinomycetes (four genera) and Bacillus (three genera). Of these isolates, Thermobifida alba strain AHK119 (AB298783) was shown to possess the ability to significantly degrade aliphatic-aromatic copolyester film as well as decreasing the polymer particle sizes when grown at 50 degrees C on LB medium supplemented with polymer particles, yielding terephthalic acid. The esterase gene (est119, 903 bp, encoding a signal peptide and a mature protein of 34 and 266 amino acids, respectively) was cloned from AHK119. The Est119 sequence contains a conserved lipase box (-G-X-S-X-G-) and a catalytic triad (Ser129, His207, and Asp175). Furthermore, Tyr59 and Met130 likely form an oxyanion hole. The recombinant enzyme was purified from cell-free extracts of Escherichia coli Rosetta-gami B (DE3) harboring pQE80L-est119. The enzyme is a monomeric protein of ca. 30 kDa, which is active from 20 degrees C to 75 degrees C (with an optimal range of 45 to 55 degrees C) and in a pH range of 5.5 to 7.0 (with an optimal pH of 6.0). Its preferred substrate among the p-nitrophenyl acyl esters (C2 to C8) is p-nitrophenyl hexanoate (C6), indicating that the enzyme is an esterase rather than a lipase.
TL;DR: In this article, Raman spectroscopy, elemental analysis and CHN determination were used to verify the presence of enzymes on supports of chitosan biocatalytic systems.
TL;DR: The objective of this work was to produce an immobilized form of lipase from Burkholderia cepacia with advantageous catalytic properties and stability to be used in the ethanolysis of different feedstocks, mainly babassu oil and tallow beef.
TL;DR: Non-ionic detergents such as Triton X-100, Tween 80 and Tween 20 strongly stimulated lipase activity, whereas SDS inhibited it and the lipase was stable in iso-octane and hexane at 80%.
TL;DR: In this article, the EC50 values for inhibition were 22μg/ml for white tea and 35μg /ml for green tea; both easily achievable from normal infusions of tea.
TL;DR: A variant of Candida antarctica lipase A (CalA) was developed for the hydrolysis of alpha-substituted p-nitrophenyl esters by directed evolution and shows (R)-selectivity, which is reversed compared to the wild type that is (S)-selective for most substrates.
Abstract: A variant of Candida antarctica lipase A (CalA) was developed for the hydrolysis of α-substituted p-nitrophenyl esters by directed evolution. The E values of this variant for 7 different esters was ...
TL;DR: The transesterification activity of the lipase from P. aeruginosa LX1 indicated that it is a potential biocatalyst for biodiesel production and higher stability in the presence of various organic solvents than in the absence of an organic solvent.
Abstract: An organic solvent-tolerant lipase from newly isolated Pseudomonas aeruginosa LX1 has been purified by ammonium sulfate precipitation and ion-exchange chromatography leading to 4.3-fold purification and 41.1% recovery. The purified lipase from P. aeruginosa LX1 was homogeneous as determined by SDS-PAGE, and the molecular mass was estimated to be 56 kDa. The optimum pH and temperature for lipase activity were found to be 7.0 and 40 °C, respectively. The lipase was stable in the pH range 4.5–12.0 and at temperatures below 50 °C. Its hydrolytic activity was found to be highest towards p-nitrophenyl palmitate (C16) among the various p-nitrophenol esters investigated. The lipase displayed higher stability in the presence of various organic solvents, such as n-hexadecane, isooctane, n-hexane, DMSO, and DMF, than in the absence of an organic solvent. The immobilized lipase was more stable in the presence of n-hexadecane, tert-butanol, and acetonitrile. The transesterification activity of the lipase from P. aeruginosa LX1 indicated that it is a potential biocatalyst for biodiesel production.
TL;DR: In this paper, the authors examined the immobilization of lipase B from Candida antarctica on functionalized multi-wall carbon nanotubes (MWCNTs) through physical adsorption.
Abstract: We examine the immobilization of lipase B from Candida antarctica on functionalized multi-wall carbon nanotubes (MWCNTs) through physical adsorption. MWCNTs functionalized with carboxyl-, amine- and ester- terminal groups on their surface are used as immobilization carriers. Dispersion of the nanotubes and the immobilization procedure take place in aqueous and low-water media. High enzyme loadings are attained, up to 25% of the weight of the carbon nanotubes. These novel biomaterials are characterized though FT-IR and Raman spectroscopy. The MWCNT–lipase bioconjugates exhibit high catalytic activity and increased storage and operational stability. The biomaterials retain more than 55% of their initial activity after 6 months at 4 °C, while they retain approximately 25% of their initial activity after 30 d of incubation in hexane at 60 °C. The catalytic behaviour of the immobilized enzyme depends on the terminal group of the carbon nanotubes, the concentration of the enzyme and the immobilization method employed.
Patent•
19 Mar 2010
TL;DR: In this paper, compositions and methods relating to lipase/acyltransferase enzymes identified in prokaryotes and eukaryotes are described, which can be used in such applications as lipid stain removal from fabrics and hard surfaces and chemical synthesis reactions.
Abstract: Compositions and methods relating to lipase/acyltransferase enzymes identified in prokaryotes and eukaryotes are described. These enzymes can be used in such applications as lipid stain removal from fabrics and hard surfaces and chemical synthesis reactions
TL;DR: In this paper, a plant enzyme with high lipase activity was extracted from germinated physic nut seeds, and applied to the production of biodiesel using a new process of enzyme/chemical hydroesterification.
Abstract: The aim of this study was to characterize a new plant enzyme (with high lipase activity) extracted from germinated physic nut seeds, and to apply this lipase to the production of biodiesel using a new process of enzyme/chemical hydroesterification. The lipase activity was detected only in the vegetable enzyme extract from germinated seeds (VEEG). Similar activities were obtained for substrates with different lengths of fatty-acid chains (111 ± 19 U/g for tributyrin (C4:0), 106 ± 49 U/g for tricaprylin (C8:0), and 96 ± 4 U/g for olive oil (C18:1)). The VEEG, obtained by a controlled processes of seed germination, was able to hydrolyze a wide range of biodiesel raw materials (vegetable oils, tallow, and biodiesel waste); of these, soy and physic nut oil showed especially high hydrolysis conversion (97% FFA). The biodiesel (fatty acid methyl esters) was produced by the hydrolysis of the physic nut oil using the VEEG, and subsequent esterification of the generated fatty acids with methanol by heterogeneous acid catalysis (niobic acid in pellets). The resulting biodiesel was of excellent quality, with the following properties: viscosity (5.5 mm 2 /s), ester content (97.1%), total glycerol (0.09 % w/w), max. methanol (0.05 % w/w), and CFPP (0 °C).
TL;DR: The combined use of Lipase AY and LipaseAK gave a higher yield of biodiesel than using Lipase AK alone, and the optimal conditions for biodiesel synthesis using mixed lipases in a batch system were: 2% water content, 10% enzyme dosage and 3:1 molar ratio of ethanol to oil.
Abstract: The enzymatic synthesis of biodiesel from used palm oil and ethanol using immobilized lipases in a solvent-free system was attempted. Five immobilized lipases, Lipase AK from Pseudomonas fluorescens, Lipase PS from Pseudomonas cepacia, Lipase AY from Candida rugosa, Lipozyme TL IM from Thermomyces lanuginosa and Novozym 435 from Candida antarctica, were screened based on their catalytic activities on reactions involved in biodiesel synthesis. The combined use of Lipase AY and Lipase AK gave a higher yield of biodiesel than using Lipase AK alone. The optimal conditions for biodiesel synthesis using mixed lipases in a batch system were: 2% water content, 10% enzyme dosage and 3:1 molar ratio of ethanol to oil. The mixed lipases could be used in 15 replicates with retained relative activity higher than 50%. In a continuous system using mixed lipases packed in packed-bed reactor, >67% of biodiesel was achieved.