Showing papers on "Lipase published in 2015"
TL;DR: It was the flavonols, not the flavanols, which showed the inhibitory activities against α-glucosidase and pancreatic lipase, thus contributing significantly to the control of blood glucose levels and obesity.
327 citations
TL;DR: An overview of recent progresses in improving IL-catalyzed biodiesel production is provided, focusing on mid- and down-stream processing such as immobilization of lipase, bioreactors development, process optimization, simulation and techno-economic evaluation.
Abstract: Transesterification of oil feedstocks using immobilized lipase (IL) is a promising process for biodiesel production. However, the running cost of this process is still higher than that of conversional chemical-catalyzed approaches. To address this challenge, both upstream and downstream processes have to be optimized. This review provides an overview of recent progresses in improving IL-catalyzed biodiesel production, focusing on mid- and down-stream processing such as immobilization of lipase, bioreactors development, process optimization, simulation and techno-economic evaluation. The immobilization of lipase is a costly process. Most of the commercial ILs are prepared by adsorption of free lipase on polymeric materials. However, to further reduce cost, works should be focused on developing cheap carriers and strengthening the interaction between enzyme and carrier but without significant loss of lipase activity. Running cost of lipase also can be reduced by improving its lifetime during transesterification. To achieve this goal, solvents can be used to prevent lipase leaching and eliminate the inhibitive effects of alcohol (usually methanol) and glycerol. Downstream processing includes important units to purify biodiesel products. In this part, works should be focused on minimizing energy consumption and waste effluents. A global process integration and optimization with economic evaluation also should be figured out to improve the economic feasibility of Il-catalyzed production of biodiesel.
300 citations
TL;DR: The purpose of this review is to give an overview of the structural and functional features of lipases and phospholipases, to describe the recent advances in optimization of the production of recombinant lipase and phosphateases, and to summarize the information available relating to their major applications in industrial processes.
Abstract: Lipases and phospholipases are interfacial enzymes that hydrolyze hydrophobic ester linkages of triacylglycerols and phospholipids, respectively. In addition to their role as esterases, these enzymes catalyze a plethora of other reactions; indeed, lipases also catalyze esterification, transesterification and interesterification reactions, and phospholipases also show acyltransferase, transacylase and transphosphatidylation activities. Thus, lipases and phospholipases represent versatile biocatalysts that are widely used in various industrial applications, such as for biodiesels, food, nutraceuticals, oil degumming and detergents; minor applications also include bioremediation, agriculture, cosmetics, leather and paper industries. These enzymes are ubiquitous in most living organisms, across animals, plants, yeasts, fungi and bacteria. For their greater availability and their ease of production, microbial lipases and phospholipases are preferred to those derived from animals and plants. Nevertheless, traditional purification strategies from microbe cultures have a number of disadvantages, which include non-reproducibility and low yields. Moreover, native microbial enzymes are not always suitable for biocatalytic processes. The development of molecular techniques for the production of recombinant heterologous proteins in a host system has overcome these constraints, as this allows high-level protein expression and production of new redesigned enzymes with improved catalytic properties. These can meet the requirements of specific industrial process better than the native enzymes. The purpose of this review is to give an overview of the structural and functional features of lipases and phospholipases, to describe the recent advances in optimization of the production of recombinant lipases and phospholipases, and to summarize the information available relating to their major applications in industrial processes.
243 citations
TL;DR: It is shown that the enzyme lipase from Rhizopus oryzae exhibits exceptionally high thermal stability and high solvent tolerance and even increased activity in acetone when immobilized onto a graphene oxide nanosupport prepared by Staudenmaier and Brodie methods.
Abstract: The thermal and solvent tolerance of enzymes is highly important for their industrial use. We show here that the enzyme lipase from Rhizopus oryzae exhibits exceptionally high thermal stability and high solvent tolerance and even increased activity in acetone when immobilized onto a graphene oxide (GO) nanosupport prepared by Staudenmaier and Brodie methods. We studied various forms of immobilization of the enzyme: by physical adsorption, covalent attachment, and additional crosslinking. The activity recovery was shown to be dependent on the support type, enzyme loading and immobilization procedure. Covalently immobilized lipase showed significantly better resistance to heat inactivation (the activity recovery was 65% at 70 °C) in comparison with the soluble counterpart (the activity recovery was 65% at 40 °C). Physically adsorbed lipase achieved over 100% of the initial activity in a series of organic solvents. These findings, showing enhanced thermal stability and solvent tolerance of graphene oxide immobilized enzyme, will have a profound impact on practical industrial scale uses of enzymes for the conversion of lipids into fuels.
180 citations
TL;DR: This review considers different solvent systems used in lipase-catalyzed reactions and some of the enzymatic properties required for function and gives lipases many potential applications in aqueous and non-aqueous biocatalysis.
150 citations
TL;DR: This article suggests that different mechanisms can lead to inactivation of different lipases, in particular substrate inhibition and protein unfolding, and attempts to improve the performances of methanol sensitive lipases by mutagenesis as well as process engineering approaches are suggested.
Abstract: The biotechnological production of biodiesel is based on transesterification/esterification reactions between a source of fatty acids and a short-chain alcohol, usually methanol, catalysed by enzymes belonging to the class known as lipases. Several lipases used in industrial processes, although stable in the presence of other organic solvents, are inactivated by methanol at or below the concentration optimal for biodiesel production, making it necessary to use stepwise methanol feeding or pre-treatment of the enzyme. In this review article we focus on what is currently know about methanol inactivation of lipases, a phenomenon which is not common to all lipase enzymes, with the goal of improving the biocatalytic process. We suggest that different mechanisms can lead to inactivation of different lipases, in particular substrate inhibition and protein unfolding. Attempts to improve the performances of methanol sensitive lipases by mutagenesis as well as process engineering approaches are also summarized.
134 citations
TL;DR: An updated review of concepts and regulation of white adipocyte lipolysis with a special emphasis in its role in metabolism homeostasis and as a source of important signaling molecules is presented.
Abstract: Lipolysis is defined as the sequential hydrolysis of triacylglycerol (TAG) stored in cell lipid droplets. For many years, it was believed that hormone-sensitive lipase (HSL) and monoacylglycerol lipase (MGL) were the main enzymes catalyzing lipolysis in the white adipose tissue. Since the discovery of adipose triglyceride lipase (ATGL) in 2004, many studies were performed to investigate and characterize the actions of this lipase, as well as of other proteins and possible regulatory mechanisms involved, which reformulated the concept of lipolysis. Novel findings from these studies include the identification of lipolytic products as signaling molecules regulating important metabolic processes in many non-adipose tissues, unveiling a previously underestimated aspect of lipolysis. Thus, we present here an updated review of concepts and regulation of white adipocyte lipolysis with a special emphasis in its role in metabolism homeostasis and as a source of important signaling molecules.
129 citations
TL;DR: While octyl-agarose released enzyme molecules after incubation at high temperatures or in the presence of organic solvents and detergents, the covalently immobilized enzyme remained attached to the support even after boiling the enzyme in SDS, eliminating the risks of product contamination.
Abstract: A new heterofunctional support, octyl-glyoxyl agarose, is proposed in this study. The supports were prepared by simple periodate oxidation of the commercial octyl-agarose, introducing 25 μmol of glyoxyl groups per wet gram of support. This support was assayed with three different lipases (those from Candida antarctica (form B), Thermomyces lanuginosus (TLL) or Rhizomucor miehei) and the artificial phospholipase Lecitase Ultra. Used at pH 7, the new support maintained as first immobilization step the lipase interfacial activation. Thus, it was possible to have the purification and immobilization of the enzyme in one step. Moreover, stabilization of the open form of the lipase was achieved. The covalent enzyme/support bonds cannot be obtained if the immobilized enzyme was not incubated at alkaline pH value. This incubation at pH 10 of the previously immobilized enzymes produced a smaller decrease in enzyme activity when compared to the direct immobilization of the enzymes on glyoxyl-agarose at pH 10, because the immobilization via interfacial activation promoted a stabilization of the lipases. Except in the case of TLL (covalent attachment involved 70% of the enzyme molecules), covalent immobilization yield was over 80%. The non-covalent attached enzyme molecules were discarded by washings with detergent solutions and the new biocatalysts were compared to the octyl-agarose immobilized enzymes. While the stability in thermal and organic solvents inactivations was increased for Lecitase Ultra, CALB and RML, TLL improved its stability in organic media but its thermal stability decreased after covalent attachment of the interfacially activated enzyme. This stabilization resulted in octyl-glyoxyl-lipase preparations which presented higher activity in the presence of organic solvents. Finally, while octyl-agarose released enzyme molecules after incubation at high temperatures or in the presence of organic solvents and detergents, the covalently immobilized enzyme remained attached to the support even after boiling the enzyme in SDS, eliminating the risks of product contamination.
122 citations
TL;DR: This review critically reports and discusses examples from recent literature, concerning the synthesis of enantiomerically pure active pharmaceutical ingredients (APIs) and their intermediates in which the key step involves the action of a lipase.
Abstract: Biocatalysis offers an alternative approach to conventional chemical processes for the production of single-isomer chiral drugs. Lipases are one of the most used enzymes in the synthesis of enantiomerically pure intermediates. The use of this type of enzyme is mainly due to the characteristics of their regio-, chemo- and enantioselectivity in the resolution process of racemates, without the use of cofactors. Moreover, this class of enzymes has generally excellent stability in the presence of organic solvents, facilitating the solubility of the organic substrate to be modified. Further improvements and new applications have been achieved in the syntheses of biologically active compounds catalyzed by lipases. This review critically reports and discusses examples from recent literature (2007 to mid-2015), concerning the synthesis of enantiomerically pure active pharmaceutical ingredients (APIs) and their intermediates in which the key step involves the action of a lipase.
114 citations
TL;DR: The findings explain the lack of interfacial activation of CALB and offer new elements to elucidate this mechanism, with the consequent implications for the catalytic properties and classification of lipases.
113 citations
TL;DR: The results show that chemical inhibition of lipase activity, genetic deficiency of adipose triglyceride lipase and, to a lesser extent, hormone-sensitive lipase blocked aP2 secretion from adipocytes, and support that targeting aP1 or the lipolysis-dependent secretory pathway may present novel mechanistic and translational opportunities in metabolic disease.
TL;DR: The results showed that the immobilization of lipase onto magnetic chitosan nanoparticles by the method of CCEE is an efficient and simple way for preparation of stable lipase.
TL;DR: The results suggest that the combined use of ultrasound and molecular sieves greatly improve esterification reactions by stabilizing the enzyme and increasing yields.
Children's Memorial Hospital1, Stanford University2, Boston Children's Hospital3, University of Amsterdam4, Columbia University5, Charles University in Prague6, University of Geneva7, University of Toronto8, Istituto Giannina Gaslini9, University of Salford10, University of Minnesota11, University of Paris12
TL;DR: This study provides the largest longitudinal case review of patients with Lal D and confirms that LAL D is predominantly a pediatric disease causing early and progressive hepatic dysfunction associated with dyslipidemia that often leads to liver failure and transplantation.
Abstract: To characterize key clinical manifestations of lysosomal acid lipase deficiency (LAL D) in children and adults.
TL;DR: A green and sustainable strategy synthesizes clinical medicine warfarin anticoagulant by using lipase-supported metal-organic framework (MOF) bioreactors (see scheme) and may be beneficial for future studies in the industrial production of chemical, pharmaceutical, and agrochemical precursors.
Abstract: A green and sustainable strategy synthesizes clinical medicine warfarin anticoagulant by using lipase- supported metal-organic framework (MOF) bioreactors (see scheme). These findings may be beneficial for future studies in the industrial production of chemical, pharma- ceutical, and agrochemical precursors.
TL;DR: This work shows that CalB displays an enhanced catalytic rate for large, bulky substrates when adsorbed to a hydrophobic interface composed of densely packed alkyl chains, and attributes this increased activity to a conformational change that yields a more open active site.
Abstract: Lipase immobilization is frequently used for altering the catalytic properties of these industrially used enzymes. Many lipases bind strongly to hydrophobic surfaces where they undergo interfacial activation. Candida antarctica lipase B (CalB), one of the most commonly used biocatalysts, is frequently discussed as an atypical lipase lacking interfacial activation. Here we show that CalB displays an enhanced catalytic rate for large, bulky substrates when adsorbed to a hydrophobic interface composed of densely packed alkyl chains. We attribute this increased activity of more than 7-fold to a conformational change that yields a more open active site. This hypothesis is supported by molecular dynamics simulations that show a high mobility for a small "lid" (helix α5) close to the active site. Molecular docking calculations confirm that a highly open conformation of this helix is required for binding large, bulky substrates and that this conformation is favored in a hydrophobic environment. Taken together, our combined approach provides clear evidence for the interfacial activation of CalB on highly hydrophobic surfaces. In contrast to other lipases, however, the conformational change only affects large, bulky substrates, leading to the conclusion that CalB acts like an esterase for small substrates and as a lipase for substrates with large alcohol substituents.
TL;DR: This review will concentrate on the role that fatty acids liberated from intracellular TAG stores play as signaling molecules, and provides an overview of the transcription factors, which are regulated by fatty acids derived from intrACEllular stores.
Abstract: Until recently, intracellular triacylglycerols (TAG) stored in the form of cytoplasmic lipid droplets have been considered to be only passive “energy conserves”. Nevertheless, degradation of TAG gives rise to a pleiotropic spectrum of bioactive intermediates, which may function as potent co-factors of transcription factors or enzymes and contribute to the regulation of numerous cellular processes. From this point of view, the process of lipolysis not only provides energy-rich equivalents but also acquires a new regulatory function. In this review, we will concentrate on the role that fatty acids liberated from intracellular TAG stores play as signaling molecules. The first part provides an overview of the transcription factors, which are regulated by fatty acids derived from intracellular stores. The second part is devoted to the role of fatty acid signaling in different organs/tissues. The specific contribution of free fatty acids released by particular lipases, hormone-sensitive lipase, adipose triacylglycerol lipase and lysosomal lipase will also be discussed.
TL;DR: The effect of nonionic, anionic, cationic, and zwitterionic detergents on the enzymatic activity and structural stability of Rhizopus niveus lipase is reported to be helpful in understanding detergent–lipase interaction in greater detail.
Abstract: In this study, we have reported the effect of nonionic, anionic, cationic, and zwitterionic detergents on the enzymatic activity and structural stability of Rhizopus niveus lipase Secondary structural changes were monitored by Far-UV CD which shows that surfactant induces helicity in the Rhizopus niveus lipase protein which was maximum in case of CTAB followed by SDS, CHAPS, and Brij-35 Similarly, tertiary structural changes were monitored by tryptophan fluorescence We also carried out enzyme kinetics assays which showed that activity was enhanced by 15- and 11-fold in the presence of CHAPS and Brij-35, respectively Furthermore, there was a decline in activity by 20 and 30 % in case of SDS and CTAB, respectively These studies may be helpful in understanding detergent–lipase interaction in greater detail as lipases are used in many industrial processes
TL;DR: The findings indicate that Lipase GH2 is a useful catalyst for conversion of microalgae oil to FAME or FAEE, and this system provides efficiency and reduced costs in biodiesel production.
TL;DR: The structural basis of Ca2+-enhanced thermostability and activity in PET-degrading cutinase-like enzyme for the first time is explained and it is found that the inactive state of Cut190S226P is activated by a conformational change in the active-site sealing residue, F106.
Abstract: A cutinase-like enzyme from Saccharomonospora viridis AHK190, Cut190, hydrolyzes the inner block of polyethylene terephthalate (PET); this enzyme is a member of the lipase family, which contains an α/β hydrolase fold and a Ser-His-Asp catalytic triad. The thermostability and activity of Cut190 are enhanced by high concentrations of calcium ions, which is essential for the efficient enzymatic hydrolysis of amorphous PET. Although Ca2+-induced thermostabilization and activation of enzymes have been well explored in α-amylases, the mechanism for PET-degrading cutinase-like enzymes remains poorly understood. We focused on the mechanisms by which Ca2+ enhances these properties, and we determined the crystal structures of a Cut190 S226P mutant (Cut190S226P) in the Ca2+-bound and free states at 1.75 and 1.45 A resolution, respectively. Based on the crystallographic data, a Ca2+ ion was coordinated by four residues within loop regions (the Ca2+ site) and two water molecules in a tetragonal bipyramidal array. Furthermore, the binding of Ca2+ to Cut190S226P induced large conformational changes in three loops, which were accompanied by the formation of additional interactions. The binding of Ca2+ not only stabilized a region that is flexible in the Ca2+-free state but also modified the substrate-binding groove by stabilizing an open conformation that allows the substrate to bind easily. Thus, our study explains the structural basis of Ca2+-enhanced thermostability and activity in PET-degrading cutinase-like enzyme for the first time and found that the inactive state of Cut190S226P is activated by a conformational change in the active-site sealing residue, F106.
TL;DR: To elucidate the kinetic mode of PL inhibition, some selected flavonoid and non-flavonoid polyphenol standards were screened for their lipase inhibition potency by their half maximal inhibitory concentration (IC50) followed by inhibition kinetic analysis.
TL;DR: It was found that lipases immobilized on silica provided biocatalyst derivatives with lower cost compared with the cost of commercially available Novozym 435 and also presented a good reusability.
TL;DR: In this article, the adsorption process was studied as a function of pH in terms of percent of adsorbed lipase, enzyme activity and zeta potential of support and enzyme.
TL;DR: The use of imidazolium-based ILs not as phase forming compounds but as adjuvants (5 wt%) in ATPS of polyethylene glycol systems with potassium phosphate buffer at pH 7 is discussed, in the extraction and purification of a lipase produced by submerged fermentation by Bacillus sp.
TL;DR: The lipase activity was strongly correlated with the formation of long-chain fatty acid esters, which could be related to the distinctive organoleptic properties of rice-koji doenjang.
Abstract: The volatile components of rice-koji doenjang produced in association with various Aspergillus species (A. oryzae, A. sojae, and A. kawachii) during 8 weeks of fermentation were compared using gas chromatography–mass spectrometry analysis and multivariate statistical analysis. In addition, the activities of diverse enzymes (α-amylase, protease, lipase, and esterase) were determined to investigate their effects on the formation of volatile compounds. Regardless of the fungi types, carbonyls including 4-methylheptan-2-one, heptan-2-one, (E)-hept-2-enal, and hexanal were found to contribute mainly to early phase fermentation, whereas the contents of ethyl esters of long-chained fatty acids were considerably enhanced in the latter stage of fermentation. With the exception of α-amylase, the activities of the studied enzymes generally increased as fermentation proceeded, and there were significant differences in enzymes activities between the species of fungi. The lipase activity was strongly correlated with th...
TL;DR: Results indicating that ABO blood type-B as well as FUT2 non-secretor status are common population-wide risk factors for developing chronic pancreatitis imply that, even within the reference range, elevated lipase activities may indicate subclinical pancreatic injury in asymptomatic subjects.
Abstract: Objective Serum lipase activities above the threefold upper reference limit indicate acute pancreatitis. We investigated whether high lipase activity—within the reference range and in the absence of pancreatitis—are associated with genetic single nucleotide polymorphisms (SNP), and whether these identified SNPs are also associated with clinical pancreatitis. Methods Genome-wide association studies (GWAS) on phenotypes ‘serum lipase activity’ and ‘high serum lipase activity’ were conducted including 3966 German volunteers from the population-based Study-of-Health-in-Pomerania (SHIP). Lead SNPs associated on a genome-wide significance level were replicated in two cohorts, 1444 blood donors and 1042 pancreatitis patients. Results Initial discovery GWAS detected SNPs within or near genes encoding the ABO blood group specifying transferases A/B ( ABO ), Fucosyltransferase-2 ( FUT2 ), and Chymotrypsinogen-B2 ( CTRB2 ), to be significantly associated with lipase activity levels in asymptomatic subjects. Replication analyses in blood donors confirmed the association of FUT-2 non-secretor status (OR=1.49; p=0.012) and ABO blood-type-B (OR=2.48; p=7.29×10 −8 ) with high lipase activity levels. In pancreatitis patients, significant associations were found for FUT-2 non-secretor status (OR=1.53; p=8.56×10 −4 ) and ABO -B (OR=1.69, p=1.0×10 −4 ) with chronic pancreatitis, but not with acute pancreatitis. Conversely, carriers of blood group O were less frequently affected by chronic pancreatitis (OR=0.62; p=1.22×10 −05) and less likely to have high lipase activity levels (OR=0.59; p=8.14×10 −05 ). Conclusions These are the first results indicating that ABO blood type-B as well as FUT2 non-secretor status are common population-wide risk factors for developing chronic pancreatitis. They also imply that, even within the reference range, elevated lipase activities may indicate subclinical pancreatic injury in asymptomatic subjects.
TL;DR: Maximum loaded Accurel-PS preparation permitted to improve the activity in this kinetic resolution compared to the PS commercial preparation by a 55-fold factor, and compared to Novozym 435 (the most active described in literature for this reaction) by a 23-fold factors.
TL;DR: In this article, a thermomyces lanuginosus lipase on Fe3O4 was studied using different covalent linkage designs, which showed high operational stability and could be simply separated by magnetization and recycled for at least 5 consecutive batches with > 80% activity remaining, suggesting its potential for application in biocatalytic biodiesel synthesis.
TL;DR: A fundamental understanding of the orientation and adsorption mechanism of lipase on four different nanomaterial surfaces with different surface chemistry are explored in detail by a combination of parallel tempering Monte Carlo (PTMC) and molecular dynamics simulations.
Abstract: Candida antarctica lipase B (CalB) is an efficient biocatalyst for hydrolysis and esterification, which plays an important role in the production of biodiesel in the bioenergy industries. The ordered immobilisation of lipases on different supports would be significant for its enzymatic catalysis in some biodiesel production processes; however, the underlying mechanisms and the preferred lipase orientation are not well understood yet. In this work, a fundamental understanding of the orientation and adsorption mechanism of lipase on four different nanomaterial surfaces with different surface chemistry are explored in detail by a combination of parallel tempering Monte Carlo (PTMC) and molecular dynamics (MD) simulations. Simulation results show that lipase is strongly adsorbed onto the hydrophobic graphite surface, as reflected by the large contact area and interaction energy; while the adsorption onto the hydrophilic TiO2 surface is weak due to two strongly adhered water layers; meanwhile lipase undergoes desorption and reorientation processes. For CalB adsorption on positively and negatively charged surfaces (NH2-SAM and COOH-SAM), the orientation distributions of lipase are narrow, and opposite orientations are obtained. CalB adsorbed on NH2-SAM has its catalytic centre oriented towards the surface, which is not conducive to the substrate binding; while the catalytic centre faces toward the solution when it is adsorbed on the COOH-SAM. Besides, the native structures of CalB adsorbed on different surfaces are preserved, which indicates lipase as a robust enzyme. The simulation results will promote our understanding on how surface properties of nanomaterials, such as charge or hydrophobicity, will affect lipase immobilisation, and help us in the rational design and development of immobilised lipase carriers.
TL;DR: Testing the enzymatic transesterification of olive and palm oils, two diverse fatty acid compositions, using standalone or mixtures of three immobilized lipases as biocatalysts showed results showing that they could be used for at least seven cycles keeping higher than 80% of their initial activities.
Abstract: Although reactions of transesterification are generally catalyzed by one specific lipase preparation, the concept of “ combi -lipase” could be better explored for the production of biodiesel, since oils are heterogeneous substrates. In this research, we tested this concept by evaluating the enzymatic transesterification of olive and palm oils, two diverse fatty acid compositions, using standalone or mixtures of three immobilized lipases as biocatalysts: Novozym 435 (CALB), Lipozyme TL-IM (TLL), and Lipozyme RM-IM (RML). For olive oil, the combination of 29.0% of TLL, 12.5% of RML, and 58.5% of CALB was the best, allowing for 95% conversion efficiency in 18 h of reaction, up from 50% for the best individual lipase (CALB). For palm oil, the best enzyme combination was 52.5% of TLL and 47.5% of RML, resulting in 80% of conversion of ethyl esters in 18 h, compared to only 44% when standalone TLL was used. Repeated batches of reaction were carried out in order to test the operational stability of the combi -lipase systems, with results showing that they could be used for at least seven cycles keeping higher than 80% of their initial activities.