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Sample records for hormone-sensitive lipase hsl

  1. Hormone-sensitive lipase (HSL) is also a retinyl ester hydrolase: evidence from mice lacking HSL.

    Science.gov (United States)

    Ström, Kristoffer; Gundersen, Thomas E; Hansson, Ola; Lucas, Stéphanie; Fernandez, Céline; Blomhoff, Rune; Holm, Cecilia

    2009-07-01

    Here, we investigated the importance of hormone-sensitive lipase (HSL) as a retinyl ester hydrolase (REH). REH activity was measured in vitro using recombinant HSL and retinyl palmitate. The expression of retinoic acid (RA)-regulated genes and retinoid metabolites were measured in high-fat diet fed HSL-null mice using real-time quantitative PCR and triple-stage liquid chromatography/tandem mass spectrometry, respectively. Age- and gender-matched wild-type littermates were used as controls. The REH activity of rat HSL was found to be higher than that against the hitherto best known HSL substrate, i.e., diacylglycerols. REH activity in white adipose tissue (WAT) of HSL-null mice was completely blunted and accompanied by increased levels of retinyl esters and decreased levels of retinol, retinaldehyde and all-trans RA. Accordingly, genes known to be positively regulated by RA were down-regulated in HSL-null mice, including pRb and RIP140, key factors promoting differentiation into the white over the brown adipocyte lineage. Dietary RA supplementation partly restored WAT mass and the expression of RA-regulated genes in WAT of HSL-null mice. These findings demonstrate the importance of HSL as an REH of adipose tissue and suggest that HSL via this action provides RA and other retinoids for signaling events that are crucial for adipocyte differentiation and lineage commitment.

  2. Human hormone-sensitive lipase (HSL): expression in white fat corrects the white adipose phenotype of HSL-deficient mice.

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    Fortier, Mélanie; Soni, Krishnakant; Laurin, Nancy; Wang, Shu Pei; Mauriège, Pascale; Jirik, Frank R; Mitchell, Grant A

    2005-09-01

    In white adipose tissue (WAT), hormone-sensitive lipase (HSL) can mediate lipolysis, a central pathway in obesity and diabetes. Gene-targeted HSL-deficient (HSL-/-) mice with no detectable HSL peptide or activity (measured as cholesteryl esterase) have WAT abnormalities, including low mass, marked heterogeneity of cell diameter, increased diacylglycerol content, and low beta-adrenergic stimulation of adipocyte lipolysis. Three transgenic mouse strains preferentially expressing human HSL in WAT were bred to a HSL-/- background. One, HSL-/- N, expresses normal human HSL (41.3 +/- 9.1% of normal activity); two express a serine-to-alanine mutant (S554A) initially hypothesized to be constitutively active: HSL-/- ML, 50.3 +/- 12.3% of normal, and HSL-/- MH, 69.8 +/- 15.8% of normal. In WAT, HSL-/- N mice resembled HSL+/+ controls in WAT mass, histology, diacylglyceride content, and lipolytic response to beta-adrenergic agents. In contrast, HSL-/- ML and HSL-/- MH mice resembled nontransgenic HSL-/- mice, except that diacylglycerol content and perirenal and inguinal WAT masses approached normal in HSL-/- MH mice. Therefore, 1) WAT expression of normal human HSL markedly improves HSL-/- WAT biochemically, physiologically, and morphologically; 2) similar levels of S554A HSL have a low physiological effect despite being active in vitro; and 3) diacylglycerol accumulation is not essential for the development of the characteristic WAT pathology of HSL-/- mice.

  3. Testosterone affects hormone-sensitive lipase (HSL) activity and lipid metabolism in the left ventricle

    DEFF Research Database (Denmark)

    Langfort, Jozef; Jagsz, Slawomir; Dobrzyn, Pawel

    2010-01-01

    Fatty acids, which are the major cardiac fuel, are derived from lipid droplets stored in cardiomyocytes, among other sources. The heart expresses hormone-sensitive lipase (HSL), which regulates triglycerides (TG) breakdown, and the enzyme is under hormonal control. Evidence obtained from adipose...... HSL activity against TG was significantly elevated in intact rats supplemented with testosterone. HSL activity against both TG and diacylglyceride was reduced by orchidectomy, whereas testosterone replacement fully reversed this effect. Moreover, testosterone increased left ventricle free fatty acid...

  4. Quarternary structure and enzymological properties of the different hormone-sensitive lipase (HSL) isoforms

    DEFF Research Database (Denmark)

    Krintel, Christian; Klint, Cecilia; Lindvall, Håkan

    2010-01-01

    Hormone-sensitive lipase (HSL) is a key enzyme in the mobilization of energy in the form of fatty acids from intracellular stores of neutral lipids. The enzyme has been shown to exist in different isoforms with different molecular masses (84 kDa, 89 kDa and 117 kDa) expressed in a tissue-dependen...

  5. Hormone-sensitive lipase (HSL) expression and regulation by epinephrine and exercise in skeletal muscle

    DEFF Research Database (Denmark)

    Ploug, Thorkil; Stallknecht, Bente Merete; Donsmark, Morten

    2002-01-01

    . Therefore, we investigated the expression and the regulation of hormone-sensitive lipase (HSL) in skeletal muscle. This enzyme is a neutral lipase and known as the rate-limiting enzyme of intracellular TG hydrolysis in adipose tissue. The total and the activated form of the neutral lipase are referred...... and contractions were partially additive. In rats training increased epinephrine-stimulated TO activity and HSL concentration in adipose tissue but not in muscle. In humans, at the end of 60 min of exercise muscle, TO activity was increased in healthy, but not in adrenalectomized, subjects. In conclusion, HSL...... in the presence of an anti-HSL antibody. The effect of epinephrine could be blocked by propanolol and mimicked by incubation of a crude supernatant from control muscle with the catalytic subunit of cAMP-dependent protein kinase. The effect of contractions was transient as TO activity declined to basal levels...

  6. Hormone-sensitive lipase (HSL) expression and regulation in skeletal muscle

    DEFF Research Database (Denmark)

    Langfort, J; Ploug, T; Ihlemann, J;

    1998-01-01

    Because the enzymatic regulation of muscle triglyceride metabolism is poorly understood we explored the character and activation of neutral lipase in muscle. Western blotting of isolated rat muscle fibers demonstrated expression of hormone-sensitive lipase (HSL). In incubated soleus muscle...... epinephrine increased neutral lipase activity by beta-adrenergic mechanisms involving cyclic AMP-dependent protein kinase (PKA). The increase was paralleled by an increase in glycogen phosphorylase activity and could be abolished by antiserum against HSL. Electrical stimulation caused a transient increase...... in activity of both neutral lipase and glycogen phosphorylase. The increase in lipase activity during contractions was not influenced by sympathectomy or propranolol. Training diminished the epinephrine induced lipase activation in muscle but enhanced the activation as well as the overall concentration...

  7. Testosterone affects hormone-sensitive lipase (HSL) activity and lipid metabolism in the left ventricle

    DEFF Research Database (Denmark)

    Langfort, Jozef; Jagsz, Slawomir; Dobrzyn, Pawel

    2010-01-01

    Fatty acids, which are the major cardiac fuel, are derived from lipid droplets stored in cardiomyocytes, among other sources. The heart expresses hormone-sensitive lipase (HSL), which regulates triglycerides (TG) breakdown, and the enzyme is under hormonal control. Evidence obtained from adipose...... tissue suggests that testosterone regulates HSL activity. To test whether this is also true in the heart, we measured HSL activity in the left ventricle of sedentary male rats that had been treated with testosterone supplementation or orchidectomy with or without testosterone substitution. Left ventricle...... HSL activity against TG was significantly elevated in intact rats supplemented with testosterone. HSL activity against both TG and diacylglyceride was reduced by orchidectomy, whereas testosterone replacement fully reversed this effect. Moreover, testosterone increased left ventricle free fatty acid...

  8. Expression of human hormone-sensitive lipase (HSL) in postmeiotic germ cells confers normal fertility to HSL-deficient mice.

    Science.gov (United States)

    Wang, Shu Pei; Chung, Shari; Soni, Krishnakant; Bourdages, Hugo; Hermo, Louis; Trasler, Jacquetta; Mitchell, Grant A

    2004-12-01

    Hormone-sensitive lipase (HSL, Lipe, E.C.3.1.1.3) is a multifunctional fatty acyl esterase that is essential for male fertility and spermatogenesis and that also plays important roles in the function of adipocytes, pancreatic beta-cells, and adrenal cortical cells. Gene-targeted HSL-deficient (HSL-/-) male mice are infertile, have a 2-fold reduction in testicular mass, a 2-fold elevation of the ratio of esterified to free cholesterol in testis, and unique morphological abnormalities in round and elongating spermatids. Postmeiotic germ cells in the testis express a specific HSL isoform. We created transgenic mice expressing a normal human testicular HSL cDNA from the mouse protamine-1 promoter, which mediates expression specifically in postmeiotic germ cells. Testicular cholesteryl esterase activity was undetectable in HSL-/- mice, but in HSL-/- males expressing the testicular transgene, activity was 2-fold greater than normal. HSL transgene mRNA became detectable in testes between 19 and 25 days of age, coinciding with the first wave of postmeiotic transcription in round spermatids. In contrast to nontransgenic HSL-/- mice, HSL-/- males expressing the testicular transgene were normal with respect to fertility, testicular mass, testicular esterified/free cholesterol ratio, and testicular histology. Their cauda epididymides contained abundant, normal-appearing spermatozoa. We conclude that human testicular HSL is functional in mouse testis and that the mechanism of infertility in HSL-deficient males is cell autonomous and resides in postmeiotic germ cells, because HSL expression in these cells is in itself sufficient to restore normal fertility.

  9. [Chromosomal localization of the hormone-sensitive lipase gene (Hsl) in rice field eel].

    Science.gov (United States)

    Ji, Fu-Yun; Yu, Qi-Xing; Pan, Pei-Wen

    2003-03-01

    Adipose tissue triacylglycerols are the quantitatively most important source of stored energy in animals. Hormone-sensitive lipase encoded by hormone-sensitive lipase gene (Hsl) is a multifunctional enzyme that catalyzes the hydrolysis of triacylglycerol stored in adipose tissue and cholesterol esters in the adrenals, ovaries, testes and macrophages. Using pig Hsl gene inserted into pBS labeled by the radioactive isotope and the digoxigenin as the probes respectively one band, 11.5kb, has been shown to hybridized with total DNA of rice field eel digested with Pst I by Southern blotting and Hsl gene has been assigned to metaphase chromosome 5, at the position of 78.35+/-1.26 from the centromere in rice field eel by fluorescent in situ hybridization (FISH). The mapping results are corresponding to that of "specific-chromosomal DNA pool" obtained by chromosome microisolation used to map gene and the mapping result is more accurate. The results of the study further illustrate the importance of the presence of Hsl gene in rice field eel genome and provide the first FISH mapping data for rice field eel chromosome 5. The current studies would advance the addition of known genetic markers and the construction of high resolution genetic map in rice field eel genome.

  10. Testosterone affects hormone-sensitive lipase (HSL) activity and lipid metabolism in the left ventricle.

    Science.gov (United States)

    Langfort, Jozef; Jagsz, Slawomir; Dobrzyn, Pawel; Brzezinska, Zofia; Klapcinska, Barbara; Galbo, Henrik; Gorski, Jan

    2010-09-01

    Fatty acids, which are the major cardiac fuel, are derived from lipid droplets stored in cardiomyocytes, among other sources. The heart expresses hormone-sensitive lipase (HSL), which regulates triglycerides (TG) breakdown, and the enzyme is under hormonal control. Evidence obtained from adipose tissue suggests that testosterone regulates HSL activity. To test whether this is also true in the heart, we measured HSL activity in the left ventricle of sedentary male rats that had been treated with testosterone supplementation or orchidectomy with or without testosterone substitution. Left ventricle HSL activity against TG was significantly elevated in intact rats supplemented with testosterone. HSL activity against both TG and diacylglyceride was reduced by orchidectomy, whereas testosterone replacement fully reversed this effect. Moreover, testosterone increased left ventricle free fatty acid levels, caused an inhibitory effect on carbohydrate metabolism in the heart, and elevated left ventricular phosphocreatine and ATP levels as compared to control rats. These data indicate that testosterone is involved in cardiac HSL activity regulation which, in turn, may affect cardiac lipid and carbohydrate metabolism.

  11. Adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (HSL) deficiencies affect expression of lipolytic activities in mouse adipose tissues.

    Science.gov (United States)

    Morak, Maria; Schmidinger, Hannes; Riesenhuber, Gernot; Rechberger, Gerald N; Kollroser, Manfred; Haemmerle, Guenter; Zechner, Rudolf; Kronenberg, Florian; Hermetter, Albin

    2012-12-01

    Adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (HSL) are key enzymes involved in intracellular degradation of triacylglycerols. It was the aim of this study to elucidate how the deficiency in one of these proteins affects the residual lipolytic proteome in adipose tissue. For this purpose, we compared the lipase patterns of brown and white adipose tissue from ATGL (-/-) and HSL (-/-) mice using differential activity-based gel electrophoresis. This method is based on activity-recognition probes possessing the same substrate analogous structure but carrying different fluorophores for specific detection of the enzyme patterns of two different tissues in one electrophoresis gel. We found that ATGL-deficiency in brown adipose tissue had a profound effect on the expression levels of other lipolytic and esterolytic enzymes in this tissue, whereas HSL-deficiency hardly showed any effect in brown adipose tissue. Neither ATGL- nor HSL-deficiency greatly influenced the lipase patterns in white adipose tissue. Enzyme activities of mouse tissues on acylglycerol substrates were analyzed as well, showing that ATGL-and HSL-deficiencies can be compensated for at least in part by other enzymes. The proteins that responded to ATGL-deficiency in brown adipose tissue were overexpressed and their activities on acylglycerols were analyzed. Among these enzymes, Es1, Es10, and Es31-like represent lipase candidates as they catalyze the hydrolysis of long-chain acylglycerols.

  12. Cloning and functional characterization of the 5' regulatory region of ovine Hormone Sensitive Lipase (HSL) gene.

    Science.gov (United States)

    Lampidonis, Antonis D; Stravopodis, Dimitrios J; Voutsinas, Gerassimos E; Messini-Nikolaki, Niki; Stefos, George C; Margaritis, Lukas H; Argyrokastritis, Alexandros; Bizelis, Iosif; Rogdakis, Emmanuel

    2008-12-31

    Hormone Sensitive Lipase (HSL) catalyzes the rate-limiting step in the mobilization of fatty acids from adipose tissue, thus determining the supply of energy substrates in the body. HSL enzymatic activity is increased by adrenergic agonists, such as catecholamines and glucagons, which induce cyclic AMP (cAMP) intracellular production, subsequently followed by the activation of Protein Kinase A (PKA) and its downstream signaling cascade reactions. HSL constitutes the critical enzyme in the modulation of lipid stores and the only component being subjected to hormonal control in terms of the recently identified Adipose Triglyceride Lipase (ATGL). In order to acquire detailed knowledge with regard to the mechanisms regulating ovine HSL (ovHSL) gene transcription activity, we initially isolated and cloned the 5' proximal and distal promoter regions through a genome walking approach, with the utilization of the already characterized ovHSL cDNAs. As evinced by BLAST analysis and a multiple alignment procedure, the isolated genomic fragment of 2.744 kb appeared to contain the already specified 5'-untranslated region (5'-UTR), which was interrupted by a relatively large intron of 1.448 kb. Regarding the upstream remaining part of 1.224 kb, it was demonstrated to represent a TATA-less promoter area, harboring several cis-regulatory elements that could be putatively recognized by relatively more general transcription factors, mainly including Stimulating protein 1 (Sp1), CCAAT-box Binding Factors (CBFs), Activator Protein 2 (AP2) and Glucocorticoid Receptor (GR), as well as other cis-acting regions denominated as Insulin Response Element (IRE), Glucose Response Element (GRE), Fat Specific Element (FSE) and cAMP Response Element (CRE), which could likely function in a nourishment (i.e. glucose)-/hormone-dependent fashion. When different genomic fragments were directionally (5' to 3') cloned into a suitable reporter vector upstream of a promoter-less luciferase gene and

  13. Cloning and functional characterization of the ovine Hormone Sensitive Lipase (HSL) full-length cDNAs: an integrated approach.

    Science.gov (United States)

    Lampidonis, Antonis D; Argyrokastritis, Alexandros; Stravopodis, Dimitrios J; Voutsinas, Gerassimos E; Ntouroupi, Triantafyllia G; Margaritis, Lukas H; Bizelis, Iosif; Rogdakis, Emmanuel

    2008-06-15

    Hormone Sensitive Lipase (HSL) is a highly regulated enzyme that mediates lipolysis in adipocytes. HSL enzymatic activity is increased by adrenergic agonists, such as catecholamines and glucagons, which induce cyclic AMP (cAMP) intracellular production, subsequently followed by the activation of Protein Kinase A (PKA) and its downstream signalling cascade reactions. Since HSL constitutes the key enzyme in the regulation of lipid stores and the only enzyme being subjected to hormonal regulation [in terms of the recently identified Adipose Triglyceride Lipase (ATGL)], the ovine Hormone Sensitive Lipase (ovHSL) full-length cDNA clones were isolated, using a Polymerase Chain Reaction-based (PCR) strategy. The two isolated isoforms ovHSL-A and ovHSL-B contain two highly homologous Open Reading Frame (ORF) regions of 2.089 Kb and 2.086 Kb, respectively, the latter having been missed the 688th triplet coding for glutamine (DeltaQ(688)). The putative 695 and 694 amino acid respective sequences bear strong homologies with other HSL protein family members. Southern blotting analysis revealed that HSL is represented as a single copy gene in the ovine genome, while Reverse Transcription-PCR (RT-PCR) approaches unambiguously dictated its variable transcriptional expression profile in the different tissues examined. Interestingly, as undoubtedly corroborated by both RT-PCR and Western blotting analysis, ovHSL gene expression is notably enhanced in the adipose tissue during the fasting period, when lipolysis is highly increased in ruminant species. Based on the crystal structure of an Archaeoglobus fulgidus enzyme, a three-dimensional (3D) molecular model of the ovHSL putative catalytic domain was constructed, thus providing an inchoative insight into understanding the enzymatic activity and functional regulation mechanisms of the ruminant HSL gene product(s).

  14. Essential role of hormone-sensitive lipase (HSL) in the maintenance of lipid storage in Mycobacterium leprae-infected macrophages.

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    Tanigawa, Kazunari; Degang, Yang; Kawashima, Akira; Akama, Takeshi; Yoshihara, Aya; Ishido, Yuko; Makino, Masahiko; Ishii, Norihisa; Suzuki, Koichi

    2012-05-01

    Mycobacterium leprae (M. leprae), the causative agent of leprosy, parasitizes within the foamy or enlarged phagosome of macrophages where rich lipids accumulate. Although the mechanisms for lipid accumulation in the phagosome have been clarified, it is still unclear how such large amounts of lipids escape degradation. To further explore underlying mechanisms involved in lipid catabolism in M. leprae-infected host cells, we examined the expression of hormone-sensitive lipase (HSL), a key enzyme in fatty acid mobilization and lipolysis, in human macrophage THP-1 cells. We found that infection by live M. leprae significantly suppressed HSL expression levels. This suppression was not observed with dead M. leprae or latex beads. Macrophage activation by peptidoglycan (PGN), the ligand for toll-like receptor 2 (TLR2), increased HSL expression; however, live M. leprae suppressed this increase. HSL expression was abolished in the slit-skin smear specimens from patients with lepromatous and borderline leprosy. In addition, the recovery of HSL expression was observed in patients who experienced a lepra reaction, which is a cell-mediated, delayed-type hypersensitivity immune response, or in patients who were successfully treated with multi-drug therapy. These results suggest that M. leprae suppresses lipid degradation through inhibition of HSL expression, and that the monitoring of HSL mRNA levels in slit-skin smear specimens may be a useful indicator of patient prognosis.

  15. The resurgence of Hormone-Sensitive Lipase (HSL) in mammalian lipolysis.

    Science.gov (United States)

    Lampidonis, Antonis D; Rogdakis, Emmanuel; Voutsinas, Gerassimos E; Stravopodis, Dimitrios J

    2011-05-15

    The ability to store energy in the form of energy-dense triacylglycerol and to mobilize these stores rapidly during periods of low carbohydrate availability or throughout the strong metabolic demand is a highly conserved process, absolutely essential for survival. In the industrialized world the regulation of this pathway is viewed as an important therapeutic target for disease prevention. Adipose tissue lipolysis is a catabolic process leading to the breakdown of triacylglycerols stored in fat cells, and release of fatty acids and glycerol. Mobilization of adipose tissue fat is mediated by the MGL, HSL and ATGL, similarly functioning enzymes. ATGL initiates lipolysis followed by the actions of HSL on diacylglycerol, and MGL on monoacylglycerol. HSL is regulated by reversible phosphorylation on five critical residues. Phosphorylation alone, however, is not enough to activate HSL. Probably, conformational alterations and a translocation from the cytoplasm to lipid droplets are also involved. In accordance, Perilipin functions as a master regulator of lipolysis, protecting or exposing the triacylglycerol core of a lipid droplet to lipases. The prototype processes of hormonal lipolytic control are the β-adrenergic stimulation and suppression by insulin, both of which affect cytoplasmic cyclic AMP levels. Lipolysis in adipocytes is an important process in the management of body energy reserves. Its deregulation may contribute to the symptoms of type 2 diabetes mellitus and other pathological situations. We, herein, discuss the metabolic regulation and function of lipases mediating mammalian lipolysis with a focus on HSL, quoting newly identified members of the lipolytic proteome.

  16. The testicular form of hormone-sensitive lipase HSLtes confers rescue of male infertility in HSL-deficient mice.

    Science.gov (United States)

    Vallet-Erdtmann, Virginie; Tavernier, Geneviève; Contreras, Juan Antonio; Mairal, Aline; Rieu, Cécile; Touzalin, Anne-Marie; Holm, Cecilia; Jégou, Bernard; Langin, Dominique

    2004-10-01

    Inactivation of the hormone-sensitive lipase gene (HSL) confers male sterility with a major defect in spermatogenesis. Several forms of HSL are expressed in testis. HSLtes mRNA and protein are found in early and elongated spermatids, respectively. The other forms are expressed in diploid germ cells and interstitial cells of the testis. To determine whether the absence of the testis-specific form of HSL, HSLtes, was responsible for the infertility in HSL-null mice, we generated transgenic mice expressing HSLtes under the control of its own promoter. The transgenic animals were crossed with HSL-null mice to produce mice deficient in HSL in nongonadal tissues but expressing HSLtes in haploid germ cells. Cholesteryl ester hydrolase activity was almost completely blunted in HSL-deficient testis. Mice with one allele of the transgene showed an increase in enzymatic activity and a small elevation in the production of spermatozoa. The few fertile hemizygous male mice produced litters of very small to small size. The presence of the two alleles led to a doubling in cholesteryl ester hydrolase activity, which represented 25% of the wild type values associated with a qualitatively normal spermatogenesis and a partial restoration of sperm reserves. The fertility of these mice was totally restored with normal litter sizes. In line with the importance of the esterase activity, HSLtes transgene expression reversed the cholesteryl ester accumulation observed in HSL-null mice. Therefore, expression of HSLtes and cognate cholesteryl ester hydrolase activity leads to a rescue of the infertility observed in HSL-deficient male mice.

  17. Variation in the ovine hormone-sensitive lipase gene (HSL) and its association with growth and carcass traits in New Zealand Suffolk sheep.

    Science.gov (United States)

    Yang, Guo; Forrest, Rachel; Zhou, Huitong; Hickford, Jonathan

    2014-01-01

    The hormone-sensitive lipase (HSL) plays an important role in the regulation of lipolysis in adipose tissues, by catalysing a rate-limiting step in triglyceride hydrolysis. Variation within the human HSL gene (HSL) has been associated with an increased risk of obesity. In this study, variation within three regions (exon 3-4, exon 5-6 and exon 9) of ovine HSL was investigated in 538 Suffolk lambs bred from 13 independent sires using PCR-SSCP. Four sequence variants of intron 5 (designated A-D) and two variants of exon 9 (designated a and b) of ovine HSL were detected. No variation was found in exon 3-4 of the gene. The associations of the variation within ovine HSL with post-weaning growth and carcass traits including eye muscle depth (EMD), eye muscle width (EMW) and fat depth above the eye muscle (FDM) were assessed in 262 of the above 538 lambs using general linear mixed-effects models. In the single variant models, the presence of intron 5 A in a lamb's genotype was associated with reduced EMD (P = 0.036) and EMW (P = 0.018), whereas the presence of intron 5 C was associated with increased EMD (P HSL intron 5 variants and post-weaning growth, or between HSL exon 9 variants, post-weaning growth or carcass traits, were found.

  18. Hormone-Sensitive Lipase Knockouts

    Directory of Open Access Journals (Sweden)

    Shen Wen-Jun

    2006-02-01

    Full Text Available Abstract All treatments for obesity, including dietary restriction of carbohydrates, have a goal of reducing the storage of fat in adipocytes. The chief enzyme responsible for the mobilization of FFA from adipose tissue, i.e., lipolysis, is thought to be hormone-sensitive lipase (HSL. Studies of HSL knockouts have provided important insights into the functional significance of HSL and into adipose metabolism in general. Studies have provided evidence that HSL, though possessing triacylglycerol lipase activity, appears to be the rate-limiting enzyme for cholesteryl ester and diacylglycerol hydrolysis in adipose tissue and is essential for complete hormone stimulated lipolysis, but other triacylglycerol lipases are important in mediating triacylglycerol hydrolysis in lipolysis. HSL knockouts are resistant to both high fat diet-induced and genetic obesity, displaying reduced quantities of white with increased amounts of brown adipose tissue, increased numbers of adipose macrophages, and have multiple alterations in the expression of genes involved in adipose differentiation, including transcription factors, markers of adipocyte differentiation, and enzymes of fatty acid and triglyceride synthesis. With disruption of lipolysis by removal of HSL, there is a drastic reduction in lipogenesis and alteration in adipose metabolism.

  19. Adipose TriGlyceride Lipase (ATGL) and Hormone-Sensitive Lipase (HSL) protein expression is decreased in the obese insulin resistant state

    NARCIS (Netherlands)

    Jocken, J.W.; Langin, D.; Smit, E.; Saris, W.H.; Valle, C.; Hul, G.B.J.; Holm, C.; Arner, P.; Blaak, E.E.

    2007-01-01

    Aim/hypothesis: Obesity is associated with increased triacylglycerol (TAG) storage in adipose tissue and insulin resistance. The mobilization of stored TAG is mediated by HSL and the recently discovered ATGL. The aim of the present study was to examine whether ATGL and HSL mRNA and protein expressio

  20. 激素敏感性脂肪酶 HSL 对生殖系统的整合调控%The Integration and Regulation of Hormone-Sensitive Lipase in Reproductive System

    Institute of Scientific and Technical Information of China (English)

    王威仪; 徐国恒

    2015-01-01

    Hormone-sensitive lipase (HSL)has long been considered as a classical rate-limiting enzyme during lipolysis since it was first described in 1 960s.HSL is regulated mainly by catecholamine,inclu-ding adrenalin.Studies in recent years indicated that the substrates for HSL are not only triglycerides, but also diacylglycerol with the catalytic activity is ten times that of triglycerides,glycerol esters and cho-lesterol esters,which overthrow the opinion that HSL is specific to triglyceride.The scientists have gener-ated HSL gene knockout mice and confirmed HSL is widely located in the reproductive system,which in-dicates that HSL may play an important role in the regulation of physiological and pathophysiological process in the reproductive system.Here,we will focus on the features of the HSL gene,mRNA and its protein,and summarize the HSL functions in the reproductive system.%激素敏感性脂肪酶被认为是经典的脂肪分解限速酶,可特异水解甘油三酯,受儿茶酚胺等激素调控。近年来研究表明:HSL 作用底物不仅有甘油三酯,还包括甘油二酯、甘油一酯、胆固醇酯等。然而,脂肪酶在生殖系统的功能并不清楚,基因敲除小鼠为证实 HSL 广泛存在于生殖系统提供良好模型,提示其可能在生殖系统生理及病理生理过程发挥重要调节作用,本文将着重介绍生殖系统中 HSL 基因与蛋白质结构并总结其在生殖系统的功能。

  1. Enhanced activity of hormone sensitive lipase (HSL) in mesenteric but not epididymal fat correlates with higher production of epinephrine in mesenteric adipocytes in rat model of cachectic rheumatoid arthritis.

    Science.gov (United States)

    Stofkova, Andrea; Krskova, Katarina; Vaculin, Simon; Jurcovicova, Jana

    2016-06-01

    Cachectic rheumatoid arthritis, the less frequent form of the disease, is associated with loss of fat mass and often more severe course of the disease. Its experimental model represents rat adjuvant arthritis (AA) characterized by edema, lack of appetite, sharp body weight and fat loss. As individual fat depots display functional differences, here we studied lipolytic activity and sensitivity to lipolytic stimuli of nodeless epididymal fat (eWAT) and perinodal mesenteric fat (mWAT) depots at the peak of AA. We also examined changes in catecholamine and cytokine levels involved in lipolysis in plasma and/or isolated adipocytes from both WATs to identify the contribution of local, adipocyte-based processes and/or systemic events to adiposity loss in cachectic rheumatoid arthritis. AA was induced to male Lewis rats by complete Freund's adjuvant. Groups of ad libitum-fed and pair-fed controls were used to distinguish the effects of food restriction from inflammation-induced cachexia. Adipose triglyceride lipase (ATGL), hormone-sensitive lipase (HSL) and its phosphorylated form (pHSL) were analyzed by western blot. CRP and catecholamine levels in plasma or adipocyte lysates were determined using ELISA kits. Cytokine-induced neutrophil chemoattractant-1 (CINC-1/CXCL1), monocyte chemoattractant protein-1 (MCP-1/CCL2), IL-1β, IL-6, IL-10 and leptin in adipocyte lysate were analyzed by quantitative protein microarray. Plasma glycerol and FFA were measured spectrophotometrically. AA rats developed severe cachexia, with lower adiposity in mWAT compared to normal and pair-fed controls, whereas in eWAT the adiposity was similarly reduced in AA and pair-fed groups. ATGL levels in both WATs were not affected by AA or pair feeding. AA upregulated levels of HSL, pHSL and pHSL/HSL ratio in mWAT, whereas none of these parameters has changed in eWAT of AA rats or in either WATs of pair-fed rats. In AA rats plasma glycerol was elevated, whereas FFA concentration was reduced. Plasma

  2. Sulforaphane induced adipolysis via hormone sensitive lipase activation, regulated by AMPK signaling pathway.

    Science.gov (United States)

    Lee, Ju-Hee; Moon, Myung-Hee; Jeong, Jae-Kyo; Park, Yang-Gyu; Lee, You-Jin; Seol, Jae-Won; Park, Sang-Youel

    2012-10-05

    Sulforaphane, an aliphatic isothiocyanate derived from cruciferous vegetables, is known for its antidiabetic properties. The effects of sulforaphane on lipid metabolism in adipocytes are not clearly understood. Here, we investigated whether sulforaphane stimulates lipolysis. Mature adipocytes were incubated with sulforaphane for 24h and analyzed using a lipolysis assay which quantified glycerol released into the medium. We investigated gene expression of hormone-sensitive lipase (HSL), and levels of HSL phosphorylation and AMP-activated protein kinase on sulforaphane-mediated lipolysis in adipocytes. Sulforaphane promoted lipolysis and increased both HSL gene expression and HSL activation. Sulforaphane suppressed AMPK phosphorylation at Thr-172 in a dose-dependent manner, which was associated with a decrease in HSL phosphorylation at Ser-565, enhancing the phosphorylation of HSL Ser-563. Taken together, these results suggest that sulforaphane promotes lipolysis via hormone sensitive lipase activation mediated by decreasing AMPK signal activation in adipocytes. Copyright © 2012 Elsevier Inc. All rights reserved.

  3. Phosphorylation of hormone-sensitive lipase by protein kinase A in vitro promotes an increase in its hydrophobic surface area

    DEFF Research Database (Denmark)

    Krintel, Christian; Mörgelin, Matthias; Logan, Derek T;

    2009-01-01

    Hormone-sensitive lipase (EC 3.1.1.79; HSL) is a key enzyme in the mobilization of fatty acids from stored triacylglycerols. HSL activity is controlled by phosphorylation of at least four serines. In rat HSL, Ser563, Ser659 and Ser660 are phosphorylated by protein kinase A (PKA) in vitro as well...

  4. [The Integration and Regulation of Hormone-Sensitive Lipase in Reproductive System].

    Science.gov (United States)

    Wang, Wei-yi; Xu, Guo-Heng

    2015-02-01

    Hormone-sensitive lipase (HSL) has long been considered as a classical rate-limiting enzyme during lipolysis since it was first described in 1960s. HSL is regulated mainly by catecholamine, including adrenalin. Studies in recent years indicated that the substrates for HSL are not only triglycerides, but also diacylglycerol with the catalytic activity is ten times that of triglycerides, glycerol esters and cholesterol esters, which overthrow the opinion that HSL is specific to triglyceride. The scientists have generated HSL gene knockout mice and confirmed HSL is widely located in the reproductive system, which indicates that HSL may play an important role in the regulation of physiological and pathophysiological process in the reproductive system. Here, we will focus on the features of the HSL gene, mRNA and its protein, and summarize the HSL functions in the reproductive system.

  5. Muscle triacylglycerol and hormone-sensitive lipase activity in untrained and trained human muscles

    DEFF Research Database (Denmark)

    Helge, Jørn Wulff; Biba, Taus O; Galbo, Henrik

    2006-01-01

    During exercise, triacylglycerol (TG) is recruited in skeletal muscles. We hypothesized that both muscle hormone-sensitive lipase (HSL) activity and TG recruitment would be higher in trained than in untrained subjects in response to prolonged exercise. Healthy male subjects (26 +/- 1 years, body ...

  6. Hormone-sensitive lipase as mediator of lipolysis in contracting skeletal muscle

    DEFF Research Database (Denmark)

    Donsmark, Morten; Langfort, Jozef; Holm, Cecilia;

    2005-01-01

    The authors propose that the enzyme hormone-sensitive lipase (HSL), which is the rate-limiting enzyme for hydrolysis of triacylglycerol in adipocytes, also regulates the intramyocellular triacylglycerol mobilization and is controlled by mechanisms similar to those regulating glycogen phosphorylase...

  7. Sex differences in hormone-sensitive lipase expression, activity, and phosphorylation in skeletal muscle at rest and during exercise

    DEFF Research Database (Denmark)

    Roepstorff, Carsten; Donsmark, Morten; Thiele, Maja

    2006-01-01

    Women have been shown to use more intramuscular triacylglycerol (IMTG) during exercise than men. To investigate whether this could be due to sex-specific regulation of hormone-sensitive lipase (HSL) and to use sex comparison as a model to gain further insight into HSL regulation, nine women and e...

  8. Contractions induce phosphorylation of the AMPK site Ser565 in hormone-sensitive lipase in muscle

    DEFF Research Database (Denmark)

    Donsmark, Morten; Langfort, Jozef; Holm, Cecilia;

    2004-01-01

    Intramyocellular triglyceride is an important energy store which is related to insulin resistance. Mobilization of fatty acids from this pool is probably regulated by hormone-sensitive lipase (HSL), which has recently been shown to exist in muscle and to be activated by epinephrine via PKA...... and by contractions via PKC and ERK. 5' AMP-activated protein kinase (AMPK) is an intracellular fuel gauge which regulates metabolism. In this study we incubated rat soleus muscle to investigate if AMPK influences HSL during 5min of repeated tetanic contractions. An eightfold increase in AMPK activity was accompanied...... by a 2.5-fold increase in phosphorylation of the AMPK-site Ser(565) in HSL (pHSL activation while HSL-Ser(565) phosphorylation was not reduced. The study indicates that during contractions AMPK phosphorylates HSL in Ser(565...

  9. Identification of a novel boronic acid as a potent, selective, and orally active hormone sensitive lipase inhibitor.

    Science.gov (United States)

    Ogiyama, Tomoko; Yamaguchi, Mitsuhiro; Kurikawa, Nobuya; Honzumi, Shoko; Yamamoto, Yuka; Sugiyama, Daisuke; Inoue, Shinichi

    2016-08-15

    Hormone sensitive lipase (HSL) is an attractive therapeutic target of dyslipidemia. We designed and synthesized several compounds as reversible HSL inhibitors with a focus on hydrophobic interactions, which was thought to be effective upon the HSL inhibitory activity. In these efforts, we identified boronated compound 12 showing a potent HSL inhibitory activity with an IC50 value of 7nM and a high selectivity against cholinesterases. Furthermore, compound 12 is the first boron containing HSL inhibitor that has shown an antilipolytic effect in rats after oral administration at 3mg/kg. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. The catalytic function of hormone-sensitive lipase is essential for fertility in male mice.

    Science.gov (United States)

    Wang, Shu Pei; Wu, Jiang Wei; Bourdages, Hugo; Lefebvre, Jean François; Casavant, Stéphanie; Leavitt, Blair R; Labuda, Damian; Trasler, Jacquetta; Smith, Charles E; Hermo, Louis; Mitchell, Grant A

    2014-08-01

    In male mice, deficiency of hormone sensitive lipase (HSL, Lipe gene, E.C.3.1.1.3) causes deficient spermatogenesis, azoospermia, and infertility. Postmeiotic germ cells express a specific HSL isoform that includes a 313 amino acid N-terminus encoded by a testis-specific exon (exon T1). The remainder of testicular HSL is identical to adipocyte HSL. The amino acid sequence of the testis-specific exon is poorly conserved, showing only a 46% amino acid identity with orthologous human and rat sequences, compared with 87% over the remainder of the HSL coding sequence, providing no evidence in favor of a vital functional role for the testis-specific N-terminus of HSL. However, exon T1 is important for Lipe transcription; in mouse testicular mRNA, we identified 3 major Lipe transcription start sites, finding numerous testicular transcription factor binding motifs upstream of the transcription start site. We directly explored two possible mechanisms for the infertility of HSL-deficient mice, using mice that expressed mutant HSL transgenes only in postmeiotic germ cells on a HSL-deficient background. One transgene expressed human HSL lacking enzyme activity but containing the testis-specific N-terminus (HSL-/-muttg mice). The other transgene expressed catalytically inactive HSL with the testis-specific N-terminal peptide (HSL-/-atg mice). HSL-/-muttg mice were infertile, with abnormal histology of the seminiferous epithelium and absence of spermatozoa in the epididymal lumen. In contrast, HSL-/-atg mice had normal fertility and normal testicular morphology. In conclusion, whereas the catalytic function of HSL is necessary for spermatogenesis in mice, the presence of the N-terminal testis-specific fragment is not essential.

  11. Adipose triglyceride lipase and hormone-sensitive lipase are the major enzymes in adipose tissue triacylglycerol catabolism.

    Science.gov (United States)

    Schweiger, Martina; Schreiber, Renate; Haemmerle, Guenter; Lass, Achim; Fledelius, Christian; Jacobsen, Poul; Tornqvist, Hans; Zechner, Rudolf; Zimmermann, Robert

    2006-12-29

    The mobilization of free fatty acids from adipose triacylglycerol (TG) stores requires the activities of triacylglycerol lipases. In this study, we demonstrate that adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (HSL) are the major enzymes contributing to TG breakdown in in vitro assays and in organ cultures of murine white adipose tissue (WAT). To differentiate between ATGL- and HSL-specific activities in cytosolic preparations of WAT and to determine the relative contribution of these TG hydrolases to the lipolytic catabolism of fat, mutant mouse models lacking ATGL or HSL and a mono-specific, small molecule inhibitor for HSL (76-0079) were used. We show that 76-0079 had no effect on TG catabolism in HSL-deficient WAT but, in contrast, essentially abolished free fatty acid mobilization in ATGL-deficient fat. CGI-58, a recently identified coactivator of ATGL, stimulates TG hydrolase activity in wild-type and HSL-deficient WAT but not in ATGL-deficient WAT, suggesting that ATGL is the sole target for CGI-58-mediated activation of adipose lipolysis. Together, ATGL and HSL are responsible for more than 95% of the TG hydrolase activity present in murine WAT. Additional known or unknown lipases appear to play only a quantitatively minor role in fat cell lipolysis.

  12. The effect of exercise training on hormone-sensitive lipase in rat intra-abdominal adipose tissue and muscle

    DEFF Research Database (Denmark)

    Enevoldsen, L H; Stallknecht, B; Langfort, J

    2001-01-01

    1. Adrenaline-stimulated lipolysis in adipose tissue may increase with training. The rate-limiting step in adipose tissue lipolysis is catalysed by the enzyme hormone-sensitive lipase (HSL). We studied the effect of exercise training on the activity of the total and the activated form of HSL......, referred to as HSL (DG) and HSL (TG), respectively, and on the concentration of HSL protein in retroperitoneal (RE) and mesenteric (ME) adipose tissue, and in the extensor digitorum longus (EDL) and soleus muscles in rats. 2. Rats (weighing 96 +/- 1 g, mean +/- S.E.M.) were either swim trained (T, 18 weeks......, n = 12) or sedentary (S, n = 12). Then RE and ME adipose tissue and the EDL and soleus muscles were incubated for 20 min with 4.4 microM adrenaline. 3. HSL enzyme activities in adipose tissue were higher in T compared with S rats. Furthermore, in RE adipose tissue, training also doubled HSL protein...

  13. Attainment of brown adipocyte features in white adipocytes of hormone-sensitive lipase null mice.

    Directory of Open Access Journals (Sweden)

    Kristoffer Ström

    Full Text Available BACKGROUND: Hormone-sensitive lipase (HSL is expressed predominantly in adipose tissue, where it plays an important role in catecholamine-stimulated hydrolysis of stored tri- and diglycerides, thus mobilizing fatty acids. HSL exhibits broad substrate specificity and besides acylglycerides it hydrolyzes cholesteryl esters, retinyl esters and lipoidal esters. Despite its role in fatty acid mobilization, HSL null mice have been shown to be resistant to diet-induced obesity. METHODOLOGY/PRINCIPAL FINDINGS: Following a high-fat diet (HFD regimen, energy expenditure, measured using indirect calorimetry, was increased in HSL null mice. White adipose tissue of HSL null mice was characterized by reduced mass and reduced protein expression of PPARgamma, a key transcription factor in adipogenesis, and stearoyl-CoA desaturase 1, the expression of which is known to be positively correlated to the differentiation state of the adipocyte. The protein expression of uncoupling protein-1 (UCP-1, the highly specific marker of brown adipocytes, was increased 7-fold in white adipose tissue of HSL null mice compared to wildtype littermates. Transmission electron microscopy revealed an increase in the size of mitochondria of white adipocytes of HSL null mice. The mRNA expression of pRb and RIP140 was decreased in isolated white adipocytes, while the expression of UCP-1 and CPT1 was increased in HSL null mice compared to wildtype littermates. Basal oxygen consumption was increased almost 3-fold in white adipose tissue of HSL null mice and was accompanied by increased uncoupling activity. CONCLUSIONS: These data suggest that HSL is involved in the determination of white versus brown adipocytes during adipocyte differentiation The exact mechanism(s underlying this novel role of HSL remains to be elucidated, but it seems clear that HSL is required to sustain normal expression levels of pRb and RIP140, which both promote differentiation into the white, rather than the brown

  14. Hormone-sensitive lipase null mice exhibit signs of impaired insulin sensitivity whereas insulin secretion is intact

    DEFF Research Database (Denmark)

    Mulder, Hindrik; Sörhede-Winzell, Maria; Contreras, Juan Antonio;

    2003-01-01

    Lipid metabolism plays an important role in glucose homeostasis under normal and pathological conditions. In adipocytes, skeletal muscle, and pancreatic beta-cells, lipids are mobilized from acylglycerides by the hormone-sensitive lipase (HSL). Here, the consequences of a targeted disruption...

  15. Ser649 and Ser650 are the major determinants of protein kinase A-mediated activation of human hormone-sensitive lipase against lipid substrates

    DEFF Research Database (Denmark)

    Krintel, Christian; Osmark, Peter; Larsen, Martin Rask;

    2008-01-01

    Hormone-sensitive lipase (HSL) is a key enzyme in the mobilization of fatty acids from stored triacylglycerols. Its activity is regulated by reversible protein phosphorylation. In rat HSL Ser563, Ser659 and Ser660 have been shown to be phosphorylated by protein kinase A (PKA) in vitro as well...

  16. Hormone-sensitive lipase-knockout mice maintain high bone density during aging.

    Science.gov (United States)

    Shen, Wen-Jun; Liu, Li-Fen; Patel, Shailja; Kraemer, Fredric B

    2011-08-01

    We tested the hypothesis that the actions of hormone-sensitive lipase (HSL) affect the microenvironment of the bone marrow and that removal of HSL function by gene deletion maintains high bone mass in aging mice. We compared littermate control wild-type (WT) and HSL(-/-) mice during aging for changes in serum biochemical values, trabecular bone density using micro-computed tomography, bone histomorphometry, and characteristics of primary bone marrow cells and preosteoblasts. There is a regulated expression of HSL and genes involved in lipid metabolism in the bone marrow during aging. HSL(-/-) mice have increased serum levels of insulin and osteocalcin with decreased leptin levels. Compared with the marked adipocyte infiltration in WT bone marrow (65% by area) at 14 mo, HSL(-/-) mice have fewer (16%, PHSL(-/-) mice maintain a higher bone density (bone volume/total volume 6.1%) with age than WT mice (2.6%, PHSL(-/-) mice show increased growth rates and higher osteogenic potential, manifested by increased expression of Runx2 (3.5-fold, PHSL directs cells within the bone marrow toward osteoblast differentiation and favors the maintenance of bone density with aging.

  17. Contraction-induced lipolysis is not impaired by inhibition of hormone-sensitive lipase in skeletal muscle

    DEFF Research Database (Denmark)

    Alsted, Thomas Junker; Ploug, Thorkil; Prats Gavalda, Clara

    2013-01-01

    In skeletal muscle hormone-sensitive lipase (HSL) has long been accepted to be the principal enzyme responsible for lipolysis of intramyocellular triacylglycerol (IMTG) during contractions. However, this notion is based on in vitro lipase activity data, which may not reflect the in vivo lipolytic...... activity. We investigated lipolysis of IMTG in soleus muscles electrically-stimulated to contract ex vivo during acute pharmacological inhibition of HSL in rat muscles and in muscles from HSL-KO mice. Measurements of IMTG are complicated by the presence of adipocytes located between the muscle fibers....... To circumvent the problem with this contamination we analyzed intramyocellular lipid droplet content histochemically. At maximal inhibition of HSL in rat muscles, contraction-induced breakdown of IMTG was identical to that seen in control muscles (p...

  18. Hormone-sensitive lipase deficiency suppresses insulin secretion from pancreatic islets of Lep{sup ob/ob} mice

    Energy Technology Data Exchange (ETDEWEB)

    Sekiya, Motohiro [Department of Metabolic Diseases, Graduate School of Medicine, University of Tokyo, Tokyo 113-8655 (Japan); Yahagi, Naoya, E-mail: nyahagi-tky@umin.ac.jp [Department of Metabolic Diseases, Graduate School of Medicine, University of Tokyo, Tokyo 113-8655 (Japan); Laboratory of Molecular Physiology on Energy Metabolism, Graduate School of Medicine, University of Tokyo, Tokyo 113-8655 (Japan); Tamura, Yoshiaki; Okazaki, Hiroaki; Igarashi, Masaki; Ohta, Keisuke; Takanashi, Mikio; Kumagai, Masayoshi; Takase, Satoru; Nishi, Makiko; Takeuchi, Yoshinori; Izumida, Yoshihiko; Kubota, Midori; Ohashi, Ken; Iizuka, Yoko [Department of Metabolic Diseases, Graduate School of Medicine, University of Tokyo, Tokyo 113-8655 (Japan); Yagyu, Hiroaki [Division of Endocrinology and Metabolism, Department of Medicine, Jichi Medical University, Tochigi 329-0498 (Japan); Gotoda, Takanari [Department of Nephrology and Endocrinology, Graduate School of Medicine, University of Tokyo, Tokyo 113-8655 (Japan); Nagai, Ryozo [Department of Cardiovascular Medicine, Graduate School of Medicine, University of Tokyo, Tokyo 113-8655 (Japan); Shimano, Hitoshi; Yamada, Nobuhiro [Advanced Biomedical Applications, Graduate School of Comprehensive Human Sciences, University of Tsukuba, Ibaragi 305-8575 (Japan); and others

    2009-09-25

    It has long been a matter of debate whether the hormone-sensitive lipase (HSL)-mediated lipolysis in pancreatic {beta}-cells can affect insulin secretion through the alteration of lipotoxicity. We generated mice lacking both leptin and HSL (Lep{sup ob/ob}/HSL{sup -/-}) and explored the role of HSL in pancreatic {beta}-cells in the setting of obesity. Lep{sup ob/ob}/HSL{sup -/-} developed elevated blood glucose levels and reduced plasma insulin levels compared with Lep{sup ob/ob}/HSL{sup +/+} in a fed state, while the deficiency of HSL did not affect glucose homeostasis in Lep{sup +/+} background. The deficiency of HSL exacerbated the accumulation of triglycerides in Lep{sup ob/ob} islets, leading to reduced glucose-stimulated insulin secretion. The deficiency of HSL also diminished the islet mass in Lep{sup ob/ob} mice due to decreased cell proliferation. In conclusion, HSL affects insulin secretary capacity especially in the setting of obesity.

  19. Contraction-induced lipolysis is not impaired by inhibition of hormone-sensitive lipase in skeletal muscle.

    Science.gov (United States)

    Alsted, Thomas J; Ploug, Thorkil; Prats, Clara; Serup, Annette K; Høeg, Louise; Schjerling, Peter; Holm, Cecilia; Zimmermann, Robert; Fledelius, Christian; Galbo, Henrik; Kiens, Bente

    2013-10-15

    In skeletal muscle hormone-sensitive lipase (HSL) has long been accepted to be the principal enzyme responsible for lipolysis of intramyocellular triacylglycerol (IMTG) during contractions. However, this notion is based on in vitro lipase activity data, which may not reflect the in vivo lipolytic activity. We investigated lipolysis of IMTG in soleus muscles electrically stimulated to contract ex vivo during acute pharmacological inhibition of HSL in rat muscles and in muscles from HSL knockout (HSL-KO) mice. Measurements of IMTG are complicated by the presence of adipocytes located between the muscle fibres. To circumvent the problem with this contamination we analysed intramyocellular lipid droplet content histochemically. At maximal inhibition of HSL in rat muscles, contraction-induced breakdown of IMTG was identical to that seen in control muscles (P contractions IMTG staining decreased significantly in both HSL-KO and WT muscles (P muscle, other TG lipases accordingly being of negligible importance for lipolysis of IMTG. The present study is the first to demonstrate that contraction-induced lipolysis of IMTG occurs in the absence of HSL activity in rat and mouse skeletal muscle. Furthermore, the results suggest that ATGL is activated and plays a major role in lipolysis of IMTG during muscle contractions.

  20. Hormone-sensitive lipase, the rate-limiting enzyme in triglyceride hydrolysis, is expressed and active in beta-cells.

    Science.gov (United States)

    Mulder, H; Holst, L S; Svensson, H; Degerman, E; Sundler, F; Ahrén, B; Rorsman, P; Holm, C

    1999-01-01

    Triglycerides in the beta-cell may be important for stimulus-secretion coupling, through provision of a lipid-derived signal, and for pathogenetic events in NIDDM, where lipids may adversely affect beta-cell function. In adipose tissues, hormone-sensitive lipase (HSL) is rate-limiting in triglyceride hydrolysis. Here, we investigated whether this enzyme is also expressed and active in beta-cells. Northern blot analysis and reverse transcription-polymerase chain reaction demonstrated that HSL is expressed in rat islets and in the clonal beta-cell lines INS-1, RINm5F, and HIT-T15. Western blot analysis identified HSL in mouse and rat islets and the clonal beta-cells. In mouse and rat, immunocytochemistry showed a predominant occurrence of HSL in beta-cells, with a presumed cytoplasmic localization. Lipase activity in homogenates of the rodent islets and clonal beta-cells constituted 2.1 +/- 0.6% of that in adipocytes; this activity was immunoinhibited by use of antibodies to HSL. The established HSL expression and activity in beta-cells offer a mechanism whereby lipids are mobilized from intracellular stores. Because HSL in adipocytes is activated by cAMP-dependent protein kinase (PKA), PKA-regulated triglyceride hydrolysis in beta-cells may participate in the regulation of insulin secretion, possibly by providing a lipid-derived signal, e.g., long-chain acyl-CoA and diacylglycerol.

  1. Hormone-sensitive lipase serine phosphorylation and glycerol exchange across skeletal muscle in lean and obese subjects

    DEFF Research Database (Denmark)

    Jocken, Johan We; Roepstorff, Carsten; Goossens, Gijs H.;

    2008-01-01

    from the vastus lateralis muscle before and during ISO to investigate hormone-sensitive lipase (HSL) protein expression and serine phosphorylation. Results: Baseline total glycerol release across the forearm was significantly blunted in obese compared with lean subjects (P=0.045). This was accompanied...... by lower HSL protein expression (P=0.004), and HSL phosphorylation on PKA sites Ser(563) (P=0.041) and Ser(659) (P=0.09) and on the AMPK site Ser(565) (P=0.007), suggesting a blunted skeletal muscle lipolysis in obesity. Total forearm glycerol uptake during baseline did not differ significantly between...... groups while higher net fatty acid uptake across the forearm was observed in the obese (P=0.064). ISO induced an increase in total glycerol release from SM, which was not significantly different between groups. Interestingly, this was accompanied by an increase in HSL Ser(659) phosphorylation in obese...

  2. Contractions activate hormone-sensitive lipase in rat muscle by protein kinase C and mitogen-activated protein kinase

    DEFF Research Database (Denmark)

    Donsmark, Morten; Langfort, Jozef; Holm, Cecilia

    2003-01-01

    Intramuscular triacylglycerol is an important energy store and is also related to insulin resistance. The mobilization of fatty acids from this pool is probably regulated by hormone-sensitive lipase (HSL), which has recently been shown to exist in muscle and to be activated by both adrenaline...... and contractions. Adrenaline acts via cAMP-dependent protein kinase (PKA). The signalling mediating the effect of contractions is unknown and was explored in this study. Incubated soleus muscles from 70 g male rats were electrically stimulated to perform repeated tetanic contractions for 5 min. The contraction...... of the inhibitors reduced adrenaline-induced HSL activation in soleus muscle. Both phorbol-12-myristate-13-acetate (PMA), which activates PKC and, in turn, ERK, and caffeine, which increases intracellular Ca2+ without eliciting contraction, increased HSL activity. Activated ERK increased HSL activity in supernatant...

  3. Data on hepatic lipolysis, adipose triglyceride lipase, and hormone-sensitive lipase in fasted and non-fasted C57BL/6J female mice

    Directory of Open Access Journals (Sweden)

    Phillip M. Marvyn

    2016-06-01

    Full Text Available Liver homogenates produced from fasted and non-fasted C57BL/6J female mice were assayed for total lipolytic activity measured as hydrolysis of [9,10-3H(N]-triolein into [3H] free fatty acids (FFA. Liver homogenates were also used for immunoblotting to determine levels of the lipolytic enzymes adipose-triglyceride lipase (ATGL and hormone-sensitive lipase (HSL, as well as site specific phosphorylation at the 14-3-3 binding site of ATGL and the serine 565 and serine 660 sites of HSL. Significantly higher triolein hydrolysis activity was observed in fasted liver samples, as well as a significant increase in total ATGL and a significant decrease in HSL phosphorylation at the S565 site.

  4. Expression of human hormone-sensitive lipase in white adipose tissue of transgenic mice increases lipase activity but does not enhance in vitro lipolysis.

    Science.gov (United States)

    Lucas, Stéphanie; Tavernier, Geneviève; Tiraby, Claire; Mairal, Aline; Langin, Dominique

    2003-01-01

    Hormone-sensitive lipase (HSL) catalyzes the hydrolysis of acylglycerols and cholesteryl esters (CEs). The enzyme is highly expressed in adipose tissues (ATs), where it is thought to play an important role in fat mobilization. The purpose of the present work was to study the effect of a physiological increase of HSL expression in vivo. Transgenic mice were produced with a 21 kb human genomic fragment encompassing the exons encoding the adipocyte form of HSL. hHSL mRNA was expressed at 3-fold higher levels than murine HSL mRNA in white adipocytes. Transgene expression was also observed in brown adipose tissue (BAT) and skeletal muscle. The human protein was detected in ATs of transgenic (Tg) mice. The hydrolytic activities against triacylglycerol (TG), diacylglycerol (DG) analog, and CE were increased in transgenic mouse AT. However, cAMP-inducible adipocyte lipolysis was lower in transgenic animals. In the B6CBA genetic background, transgenic mice up to 14 weeks of age showed lower body weight and fat mass. The phenotype was not observed in older animals and in mice fed a high-fat diet (HFD). In the OF1 genetic background, there was no difference in fat mass of mice fed ad libitum. However, transgenic mice became leaner than their wild-type (WT) littermates after a 4 day calorie restriction. The data show that overexpression of HSL, despite increased lipase activity, does not lead to enhanced lipolysis.

  5. Different effects of growth hormone and fasting on the induction patterns of two hormone-sensitive lipase genes in red seabream Pagrus major.

    Science.gov (United States)

    Khieokhajonkhet, Anurak; Kaneko, Gen; Hirano, Yuki; Wang, Lu; Ushio, Hideki

    2016-09-15

    Growth hormone (GH) increases phosphorylation and mRNA levels of hormone-sensitive lipase (HSL) in the livers of some marine teleosts. The hepatic GH-HSL axis appears to play important roles in fasting-induced lipolysis. However, it is not known whether GH exerts similar effects on HSL in fish adipose tissues. Functional differentiation of two fish-specific HSL isoforms (HSL1 and HSL2) also remains unclear. The present study seeks to address two unanswered questions about fish lipolysis using red seabream (Pagrus major): (1) Does GH increase phosphorylation and mRNA levels of HSL in adipose tissue? (2) How do GH and fasting affect mRNA levels of two HSL isoform genes in the liver and adipose tissue? To this end, we first cloned HSL1 and HSL2 cDNAs and investigated their tissue distribution. Transcripts of both HSLs and HSL1 proteins were abundant in the visceral adipose tissue, gonads, and liver, suggesting the important role of HSL in adipose tissue lipolysis. HSL2 transcript levels were 20-65% those of HSL1 except in the skin, and HSL2 proteins were not detected by our in-house antisera. Ex vivo administration of GH increased HSL1 phosphorylation, non-esterified fatty acid (NEFA) release, and levels of HSL1 and HSL2 mRNA in both the liver and visceral adipose tissue. Hepatic HSL2 mRNA was particularly sensitive to GH administration and sometimes exceeded HSL1 mRNA levels with up to 13-fold induction. In contrast, fasting for 4 and 7d increased HSL1 mRNA levels, but had only marginal effects on HSL2 mRNA levels in both adipose tissue or liver. We concluded that GH would increase HSL mRNAs during adipose tissue lipolysis in red seabream; however, GH and fasting result in different induction ratio of two HSL isoform genes, suggesting that other hormone(s) also contributes to fasting-induced lipolysis. Copyright © 2016 Elsevier Inc. All rights reserved.

  6. Regulation and role of hormone-sensitive lipase in rat skeletal muscle

    DEFF Research Database (Denmark)

    Donsmark, Morten; Langfort, Jozef; Holm, Cecilia

    2004-01-01

    fibre types, being higher in oxidative fibres than in glycolytic fibres. When analysed under conditions optimal for HSL, neutral lipase activity in muscle can be stimulated by adrenaline as well as by contractions. These increases are abolished by the presence of anti-HSL antibody during analysis....... Moreover, immunoprecipitation with affinity-purified anti-HSL antibody causes similar reductions in muscle HSL protein concentration and in measured neutral lipase responses to contractions. The immunoreactive HSL in muscle is stimulated by adrenaline via beta-adrenergic activation of c...... of ERK is not necessary for stimulation of HSL by adrenaline. However, contraction-induced HSL activation is mediated by PKC, at least partly via the ERK pathway. In fat cells ERK is known to phosphorylate HSL at Ser(600). Hence, phosphorylation of different sites may explain the finding that in muscle...

  7. Mechanisms of action of hormone-sensitive lipase in mouse Leydig cells: its role in the regulation of the steroidogenic acute regulatory protein.

    Science.gov (United States)

    Manna, Pulak R; Cohen-Tannoudji, Joëlle; Counis, Raymond; Garner, Charles W; Huhtaniemi, Ilpo; Kraemer, Fredric B; Stocco, Douglas M

    2013-03-22

    Hormone-sensitive lipase (HSL) catalyzes the hydrolysis of cholesteryl esters in steroidogenic tissues and, thus, facilitates cholesterol availability for steroidogenesis. The steroidogenic acute regulatory protein (StAR) controls the rate-limiting step in steroid biosynthesis. However, the modes of action of HSL in the regulation of StAR expression remain obscure. We demonstrate in MA-10 mouse Leydig cells that activation of the protein kinase A (PKA) pathway, by a cAMP analog Bt2cAMP, enhanced expression of HSL and its phosphorylation (P) at Ser-660 and Ser-563, but not at Ser-565, concomitant with increased HSL activity. Phosphorylation and activation of HSL coincided with increases in StAR, P-StAR (Ser-194), and progesterone levels. Inhibition of HSL activity by CAY10499 effectively suppressed Bt2cAMP-induced StAR expression and progesterone synthesis. Targeted silencing of endogenous HSL, with siRNAs, resulted in increased cholesteryl ester levels and decreased cholesterol content in MA-10 cells. Depletion of HSL affected lipoprotein-derived cellular cholesterol influx, diminished the supply of cholesterol to the mitochondria, and resulted in the repression of StAR and P-StAR levels. Cells overexpressing HSL increased the efficacy of liver X receptor (LXR) ligands on StAR expression and steroid synthesis, suggesting HSL-mediated steroidogenesis entails enhanced oxysterol production. Conversely, cells deficient in LXRs exhibited decreased HSL responsiveness. Furthermore, an increase in HSL was correlated with the LXR target genes, steroid receptor element-binding protein 1c and ATP binding cassette transporter A1, demonstrating HSL-dependent regulation of steroidogenesis predominantly involves LXR signaling. LXRs interact/cooperate with RXRs and result in the activation of StAR gene transcription. These findings provide novel insight and demonstrate the molecular events by which HSL acts to drive cAMP/PKA-mediated regulation of StAR expression and

  8. Structural and functional analysis of a novel EstE5 belonging to the subfamily of hormone-sensitive lipase.

    Science.gov (United States)

    Nam, Ki Hyun; Kim, Min-Young; Kim, Soo-Jin; Priyadarshi, Amit; Lee, Won Ho; Hwang, Kwang Yeon

    2009-02-06

    Hormone-sensitive lipase (HSL) plays an important role in the regulation of rodent fat cell lipolysis. It is regarded as an adipose tissue-specific enzyme whose sole metabolic role is the catalysis of hormone-stimulated lipolysis in mammalian cells. In this report we describe the functional and structural analysis of an EstE5 protein from a soil metagenome library. Function analysis results indicated that EstE5 preferentially hydrolyzes short-chain ester compounds, and our kinetic studies revealed the optimal pH and temperature. Based on the structural analysis, we defined the active site and the binding pocket. Structurally, EstE5 belongs to the HSL family and these structural studies may have applications in the production of value-added products, including pharmaceuticals.

  9. Phenytoin alters transcript levels of hormone-sensitive lipase in muscle from horses with hyperkalemic periodic paralysis.

    Science.gov (United States)

    Yudkowsky, M L; Beech, J; Fletcher, J E

    1998-10-15

    In equine hyperkalemic periodic paralysis (HyperPP), there is evidence suggesting that the primary defect in the sodium channel is associated with a secondary alteration in triacylglycerol-associated fatty acid metabolism (TAFAM) in skeletal muscle. Furthermore, TAFAM may be involved in the therapeutic action of phenytoin. The effects of phenytoin treatment on the transcript levels of three key proteins in TAFAM, hormone sensitive lipase (HSL), carnitine palmitoyltransferase (CPT), and fatty acid binding protein (FABP), were examined. These transcripts were quantitated by competitive reverse transcription polymerase chain reaction in undifferentiated and differentiated primary cultures of equine skeletal muscle from control, heterozygous HyperPP, and homozygous-affected HyperPP horses. There was a 10-fold lower level of HSL transcript in both undifferentiated and differentiated cultures from homozygous-affected horses than from horses of the other genotypes. Phenytoin selectively increased the HSL transcript in homozygous-affected differentiated cultures to levels similar to those of the other genotypes. The levels of CPT and FABP transcripts were unaffected by genotype, differentiation, and phenytoin treatment. These results suggest that the primary defect in HyperPP may secondarily decrease HSL transcript levels and that the therapeutic action of phenytoin may include regulation of mRNA transcripts in skeletal muscle. Copyright 1998 Academic Press.

  10. Reactive oxygen species facilitate translocation of hormone sensitive lipase to the lipid droplet during lipolysis in human differentiated adipocytes.

    Directory of Open Access Journals (Sweden)

    Sarah A Krawczyk

    Full Text Available In obesity, there is an increase in reactive oxygen species (ROS within adipose tissue caused by increases in inflammation and overnutrition. Hormone sensitive lipase (HSL is part of the canonical lipolytic pathway and critical for complete lipolysis. This study hypothesizes that ROS is a signal that integrates regulation of lipolysis by targeting HSL. Experiments were performed with human differentiated adipocytes from the subcutaneous depot. Antioxidants were employed as a tool to decrease ROS, and it was found that scavenging ROS with diphenyliodonium, N-acetyl cysteine, or resveratrol decreased lipolysis in adipocytes. HSL phosphorylation of a key serine residue, Ser552, as well as translocation of this enzyme from the cytosol to the lipid droplet upon lipolytic stimulation were both abrogated by scavenging ROS. The phosphorylation status of other serine residues on HSL were not affected. These findings are significant because they document that ROS contributes to the physiological regulation of lipolysis via an effect on translocation. Such regulation could be useful in developing new obesity therapies.

  11. Reactive oxygen species facilitate translocation of hormone sensitive lipase to the lipid droplet during lipolysis in human differentiated adipocytes.

    Science.gov (United States)

    Krawczyk, Sarah A; Haller, Jorge F; Ferrante, Tom; Zoeller, Raphael A; Corkey, Barbara E

    2012-01-01

    In obesity, there is an increase in reactive oxygen species (ROS) within adipose tissue caused by increases in inflammation and overnutrition. Hormone sensitive lipase (HSL) is part of the canonical lipolytic pathway and critical for complete lipolysis. This study hypothesizes that ROS is a signal that integrates regulation of lipolysis by targeting HSL. Experiments were performed with human differentiated adipocytes from the subcutaneous depot. Antioxidants were employed as a tool to decrease ROS, and it was found that scavenging ROS with diphenyliodonium, N-acetyl cysteine, or resveratrol decreased lipolysis in adipocytes. HSL phosphorylation of a key serine residue, Ser552, as well as translocation of this enzyme from the cytosol to the lipid droplet upon lipolytic stimulation were both abrogated by scavenging ROS. The phosphorylation status of other serine residues on HSL were not affected. These findings are significant because they document that ROS contributes to the physiological regulation of lipolysis via an effect on translocation. Such regulation could be useful in developing new obesity therapies.

  12. Inhibition of hormone-sensitive lipase gene expression by cAMP and phorbol esters in 3T3-F442A and BFC-1 adipocytes.

    Science.gov (United States)

    Plée-Gautier, E; Grober, J; Duplus, E; Langin, D; Forest, C

    1996-09-15

    Hormone-sensitive lipase (HSL) catalyses the rate-limiting step in adipocyte lipolysis. Short-term hormonal regulation of HSL activity is well characterized, whereas little is known about the control of HSL gene expression. We have measured HSL mRNA content of 3T3-F442A and BFC-1 adipocytes in response to the cAMP analogue 8-(4-chlorophenylthio)-cAMP (8-CPT-cAMP) and to the phorbol ester phorbol 12-myristate 13-acetate (PMA) by Northern blot, using a specific mouse cDNA fragment. Treatment of the cells for 12 or 6 h with, respectively, 0.5 mM 8-CPT-cAMP or 1 microM PMA produced a maximal decrease of about 60% in HSL mRNA. These effects were unaffected by the protein-synthesis inhibitor anisomycin, suggesting that cAMP and PMA actions were direct. The reduction in HSL mRNA was accompanied by a reduction in HSL total activity. The intracellular routes that cAMP and PMA follow for inducing such an effect seemed clearly independent. (i) After desensitization of the protein kinase C regulation pathway by a 24 h treatment of the cells with 1 microM PMA, PMA action was abolished whereas cAMP was still fully active. (ii) Treatment with saturating concentrations of both agents produced an additive effect. (iii) The synthetic glucocorticoid dexamethasone had no proper effect on HSL gene expression but potentiated cAMP action without affecting PMA action. cAMP inhibitory action on HSL is unexpected. Indeed, the second messenger of catecholamines is the main activator of HSL by phosphorylation. We envision that a long-term cAMP treatment of adipocytes induces a counter-regulatory process that reduces HSL content and, ultimately, limits fatty acid depletion from stored triacylglycerols.

  13. Parathyroid hormone induces adipocyte lipolysis via PKA-mediated phosphorylation of hormone-sensitive lipase.

    Science.gov (United States)

    Larsson, Sara; Jones, Helena A; Göransson, Olga; Degerman, Eva; Holm, Cecilia

    2016-03-01

    Parathyroid hormone (PTH) is secreted from the parathyroid glands in response to low plasma calcium levels. Besides its classical actions on bone and kidney, PTH may have other important effects, including metabolic effects, as suggested for instance by increased prevalence of insulin resistance and type 2 diabetes in patients with primary hyperparathyroidism. Moreover, secondary hyperparathyroidism may contribute to the metabolic derangements that characterize states of vitamin D deficiency. PTH has been shown to induce adipose tissue lipolysis, but the details of the lipolytic action of PTH have not been described. Here we used primary mouse adipocytes to show that intact PTH (1-84) as well as the N-terminal fragment (1-37) acutely stimulated lipolysis in a dose-dependent manner, whereas the C-terminal fragment (38-84) was without lipolytic effect. The lipolytic action of PTH was paralleled by phosphorylation of known protein kinase A (PKA) substrates, i.e. hormone-sensitive lipase (HSL) and perilipin. The phosphorylation of HSL in response to PTH occurred at the known PKA sites S563 and S660, but not at the non-PKA site S565. PTH-induced lipolysis, as well as phosphorylation of HSL at S563 and S660, was blocked by both the PKA-inhibitor H89 and the adenylate cyclase inhibitor MDL-12330A, whereas inhibitors of extracellular-regulated kinase (ERK), protein kinase B (PKB), AMP-activated protein kinase (AMPK) and Ca(2+)/calmodulin-dependent protein kinase (CaMK) had little or no effect. Inhibition of phosphodiesterase 4 (PDE4) strongly potentiated the lipolytic action of PTH, whereas inhibition of PDE3 had no effect. Our results show that the lipolytic action of PTH is mediated by the PKA signaling pathway with no or minor contribution of other signaling pathways and, furthermore, that the lipolytic action of PTH is limited by simultaneous activation of PDE4. Knowledge of the signaling pathways involved in the lipolytic action of PTH is important for our

  14. Pancreatic beta-cell lipotoxicity induced by overexpression of hormone-sensitive lipase

    DEFF Research Database (Denmark)

    Winzell, Maria Sörhede; Svensson, Håkan; Enerbäck, Sven

    2003-01-01

    Lipid perturbations associated with triglyceride overstorage in beta-cells impair insulin secretion, a process termed lipotoxicity. To assess the role of hormone-sensitive lipase, which is expressed and enzymatically active in beta-cells, in the development of lipotoxicity, we generated transgenic...... mice overexpressing hormone-sensitive lipase specifically in beta-cells. Transgenic mice developed glucose intolerance and severely blunted glucose-stimulated insulin secretion when challenged with a high-fat diet. As expected, both lipase activity and forskolin-stimulated lipolysis was increased...... results highlight the importance of mobilization of the islet triglyceride pool in the development of beta-cell lipotoxicity. We propose that hormone-sensitive lipase is involved in mediating beta-cell lipotoxicity by providing ligands for peroxisome proliferator-activated receptors and other lipid...

  15. Depot-specific differences in perilipin and hormone-sensitive lipase expression in lean and obese

    Directory of Open Access Journals (Sweden)

    Beylot Michel

    2009-12-01

    Full Text Available Abstract Background Mainly dependent on hormone-sensitive lipase, lipolysis is differently impaired between fat depots in human obesity. Perilipin A expression is a critical element in adipocyte lipolysis. The present study aimed at comparing expression and subcellular distribution of perilipin and hormone-sensitive lipase in two abdominal adipose tissues of lean and obese women. We examined whether regional differences in perilipin expression contribute to impaired lipolytic rates. Methods Abdominal subcutaneous and omental adipose tissues were obtained from six lean and ten obese women. We measured total protein content and relative distribution of hormone-sensitive lipase and perilipin proteins between lipid and non-lipid fractions in tissue homogenates. Hormone-sensitive lipase and perilipin mRNA levels, adipocyte size, basal (non-stimulated and noradrenaline-stimulated lipolysis in isolated adipocytes were determined. Results Adipocytes were significantly larger in the obese versus the lean women and in subcutaneous versus omental fat. Expressed as a function of cell number, basal lipolysis and noradrenaline responsiveness were higher in subcutaneous versus omental adipocytes from the obese women (P Conclusion In both fat depots, a reduced perilipin protein expression was observed in women obesity. Perilipin protein level may contribute to differences in basal lipolysis and in adipocyte size between fat depots and may regulate lipid accumulation in adipocytes. Differences in hormone-sensitive lipase subcellular distribution were reported between fat depots in the obese women.

  16. The molecular mechanism of human hormone-sensitive lipase inhibition by substituted 3-phenyl-5-alkoxy-1,3,4-oxadiazol-2-ones.

    Science.gov (United States)

    Ali, Yassine Ben; Verger, Robert; Carrière, Frédéric; Petry, Stefan; Muller, Günter; Abousalham, Abdelkarim

    2012-01-01

    Hormone-sensitive lipase (HSL) plays an important role in the mobilization of free fatty acids (FFA) from adipocytes. The inhibition of HSL may offer a pharmacological approach to reduce FFA levels in plasma and diminish peripheral insulin resistance in type 2 diabetes. In this work, the inhibition of HSL by substituted 3-phenyl-5-alkoxy-1,3,4-oxadiazol-2-ones has been studied in vitro. 5-methoxy-3-(3-phenoxyphenyl)-1,3,4-oxadiazol-2(3H)-one (compound 7600) and 5-methoxy-3-(3-methyl-4-phenylacetamidophenyl)-1,3,4-oxadiazol-2(3H)-one (compound 9368) were selected as the most potent HSL inhibitors. HSL is inhibited after few minutes of incubation with compound 7600, at a molar excess of 20. This inhibition is reversed in the presence of an emulsion of lipid substrate. The reactivation phenomenon is hardly observed when incubating HSL with compound 9368. The molecular mechanism underlying the reversible inhibition of HSL by compound 7600 was investigated using high performance liquid chromatography and tandem mass spectrometry. The stoichiometry of the inhibition reaction revealed that specifically one molecule of inhibitor was bound per enzyme molecule. The inhibition by compound 7600 involves a nucleophilic attack by the hydroxy group of the catalytic Ser of the enzyme on the carbon atom of the carbonyl moiety of the oxadiazolone ring of the inhibitor, leading to the formation of covalent enzyme-inhibitor intermediate. This covalent intermediate is subsequently hydrolyzed, releasing an oxadiazolone decomposition product, carbon dioxide and the active HSL form. On the basis of this study, a kinetic model is proposed to describe the inhibition of HSL by compound 7600 in the aqueous phase as well as its partial reactivation at the lipid-water interface.

  17. Substrate specificity and kinetic properties of enzymes belonging to the hormone-sensitive lipase family: comparison with non-lipolytic and lipolytic carboxylesterases.

    Science.gov (United States)

    Chahinian, Henri; Ali, Yassine Ben; Abousalham, Abdelkarim; Petry, Stefan; Mandrich, Luigi; Manco, Guiseppe; Canaan, Stephane; Sarda, Louis

    2005-12-30

    We have studied the kinetics of hydrolysis of triacylglycerols, vinyl esters and p-nitrophenyl butyrate by four carboxylesterases of the HSL family, namely recombinant human hormone-sensitive lipase (HSL), EST2 from Alicyclobacillus acidocaldarius, AFEST from Archeoglobus fulgidus, and protein RV1399C from Mycobacterium tuberculosis. The kinetic properties of enzymes of the HSL family have been compared to those of a series of lipolytic and non-lipolytic carboxylesterases including human pancreatic lipase, guinea pig pancreatic lipase related protein 2, lipases from Mucor miehei and Thermomyces lanuginosus, cutinase from Fusarium solani, LipA from Bacillus subtilis, porcine liver esterase and Esterase A from Aspergilus niger. Results indicate that human HSL, together with other lipolytic carboxylesterases, are active on short chain esters and hydrolyze water insoluble trioctanoin, vinyl laurate and olive oil, whereas the action of EST2, AFEST, protein RV1399C and non-lipolytic carboxylesterases is restricted to solutions of short chain substrates. Lipolytic and non-lipolytic carboxylesterases can be differentiated by their respective value of K(0.5) (apparent K(m)) for the hydrolysis of short chain esters. Among lipolytic enzymes, those possessing a lid domain display higher activity on tributyrin, trioctanoin and olive oil suggesting, then, that the lid structure contributes to enzyme binding to triacylglycerols. Progress reaction curves of the hydrolysis of p-nitrophenyl butyrate by lipolytic carboxylesterases with lid domain show a latency phase which is not observed with human HSL, non-lipolytic carboxylesterases, and lipolytic enzymes devoid of a lid structure as cutinase.

  18. Pancreatic beta-cell lipotoxicity induced by overexpression of hormone-sensitive lipase

    DEFF Research Database (Denmark)

    Winzell, Maria Sörhede; Svensson, Håkan; Enerbäck, Sven

    2003-01-01

    Lipid perturbations associated with triglyceride overstorage in beta-cells impair insulin secretion, a process termed lipotoxicity. To assess the role of hormone-sensitive lipase, which is expressed and enzymatically active in beta-cells, in the development of lipotoxicity, we generated transgenic...... mice overexpressing hormone-sensitive lipase specifically in beta-cells. Transgenic mice developed glucose intolerance and severely blunted glucose-stimulated insulin secretion when challenged with a high-fat diet. As expected, both lipase activity and forskolin-stimulated lipolysis was increased......-activated transcription factors, which in turn alter the expression of critical genes. One such gene might be uncoupling protein-2, which was found to be upregulated in transgenic islets, a change that was accompanied by decreased ATP levels....

  19. Endothelin regulates intermittent hypoxia-induced lipolytic remodelling of adipose tissue and phosphorylation of hormone-sensitive lipase.

    Science.gov (United States)

    Briançon-Marjollet, Anne; Monneret, Denis; Henri, Marion; Hazane-Puch, Florence; Pepin, Jean-Louis; Faure, Patrice; Godin-Ribuot, Diane

    2016-03-15

    Obstructive sleep apnoea syndrome is characterized by repetitive episodes of upper airway collapse during sleep resulting in chronic intermittent hypoxia (IH). Obstructive sleep apnoea syndrome, through IH, promotes cardiovascular and metabolic disorders. Endothelin-1 (ET-1) secretion is upregulated by IH, and is able to modulate adipocyte metabolism. Therefore, the present study aimed to characterize the role of ET-1 in the metabolic consequences of IH on adipose tissue in vivo and in vitro. Wistar rats were submitted to 14 days of IH-cycles (30 s of 21% FiO2 and 30 s of 5% FiO2 ; 8 h day(-1) ) or normoxia (air-air cycles) and were treated or not with bosentan, a dual type A and B endothelin receptor (ETA-R and ETB-R) antagonist. Bosentan treatment decreased plasma free fatty acid and triglyceride levels, and inhibited IH-induced lipolysis in adipose tissue. Moreover, IH induced a 2-fold increase in ET-1 transcription and ETA-R expression in adipose tissue that was reversed by bosentan. In 3T3-L1 adipocytes, ET-1 upregulated its own and its ETA-R transcription and this effect was abolished by bosentan. Moreover, ET-1 induced glycerol release and inhibited insulin-induced glucose uptake. Bosentan and BQ123 inhibited these effects. Bosentan also reversed the ET-1-induced phosphorylation of hormone-sensitive lipase (HSL) on Ser(660) . Finally, ET-1-induced lipolysis and HSL phosphorylation were also observed under hypoxia. Altogether, these data suggest that ET-1 is involved in IH-induced lipolysis in Wistar rats, and that upregulation of ET-1 production and ETA-R expression by ET-1 itself under IH could amplify its effects. Moreover, ET-1-induced lipolysis could be mediated through ETA-R and activation of HSL by Ser(660) phosphorylation. © 2015 The Authors. The Journal of Physiology © 2015 The Physiological Society.

  20. Adiponectin Effect of ADPN mRNA,Hormone-sensitive Lipase mRNA Expression on Primary Cultured Bovine Adipocytes%脂联素对体外培养新生牛脂肪细胞ADPN、HSL mRNA表达的影响

    Institute of Scientific and Technical Information of China (English)

    于国健; 刘国文; 李小兵; 王哲; 李鹏; 黄海龙; 张燚

    2011-01-01

    This experiement on adipocyte of calf was adopt to make primary culture, select monolayer growth well adipocyte of calf, then add concentration of the additives was as ADPN: 0,2. 5,5,15,30,50,100 μg/mL, after 12 h cultivation, abstract total RNA and test the effect of ADPN on the fat cell of ADPN mRNA and HSL mRNA expression by FQ-PCR method. With increasing the addition of ADPN, expression level of ADPN mRNA decreased; the expression level of HSL mRNA in fat cell was highest when the concentration of ADPN was 15 μg/mL in cell culture media,then decreased. The results showed as follows: the ADPN mRNA expression was regulated by negative regulator itself in adipocyte culture in vitro, but the appropriate amount concentration of ADPN could stimulate adipocyte HSL mRNA expression.%取单层生长状态良好的犊牛脂肪细胞,分别添加0、2.5、5、15、30、50、100 μg/mL外源牛重组脂联素(adiponectin,ADPN),培养12 h后提取总RNA,采用荧光定量PCR方法检测外源脂联素对脂肪细胞ADPN mRNA和HSL mRNA表达水平的影响.结果显示,随着ADPN添加量的增多,ADPN mRNA相对表达量呈下降趋势;添加ADPN 15 μg/mL时,HSLmRNA的表达量达到峰值,之后呈下降趋势.结果表明体外培养的脂肪细胞中ADPN mRNA表达量受自身的负调节,而添加适量的ADPN可刺激脂肪细胞HSL mRNA的表达.

  1. Reduced lipolysis response to adipose afferent reflex involved in impaired activation of adrenoceptor-cAMP-PKA-hormone sensitive lipase pathway in obesity

    Science.gov (United States)

    Ding, Lei; Zhang, Feng; Zhao, Ming-Xia; Ren, Xing-Sheng; Chen, Qi; Li, Yue-Hua; Kang, Yu-Ming; Zhu, Guo-Qing

    2016-01-01

    Chemical stimulation of white adipose tissue (WAT) causes adipose afferent reflex (AAR) and sympathetic activation. This study is to investigate the effects of AAR on lipolysis and the mechanisms of attenuated lipolysis response to enhanced AAR in obesity. Obesity was caused by high-fat diet for 12 weeks in rats. AAR was induced by injection of capsaicin into inguinal WAT or electrical stimulation of epididymal WAT afferent nerve. AAR caused sympathetic activation, which was enhanced in obesity rats. AAR increased cAMP levels and PKA activity, promoted hormone sensitive lipase (HSL) and perilipin phosphorylation, and increased lipolysis in WAT, which were attenuated in obesity rats. PKA activity, cAMP, perilipin and β-adrenoceptor levels were reduced, while HSL was upregulated in adipocytes from obesity rats. In primary adipocytes, isoproterenol increased cAMP levels and PKA activity, promoted HSL and perilipin phosphorylation, and increased lipolysis, which were attenuated in obesity rats. The attenuated effects of isoproterenol in adipocytes from obesity rats were prevented by a cAMP analogue dbcAMP. The results indicate that reduced lipolysis response to enhanced AAR in obesity is attributed to the impaired activation of β-adrenoceptor-cAMP-PKA-HSL pathway. Increased cAMP level in adipocytes rectifies the attenuated lipolysis in obesity. PMID:27694818

  2. Reduced lipolysis response to adipose afferent reflex involved in impaired activation of adrenoceptor-cAMP-PKA-hormone sensitive lipase pathway in obesity.

    Science.gov (United States)

    Ding, Lei; Zhang, Feng; Zhao, Ming-Xia; Ren, Xing-Sheng; Chen, Qi; Li, Yue-Hua; Kang, Yu-Ming; Zhu, Guo-Qing

    2016-10-03

    Chemical stimulation of white adipose tissue (WAT) causes adipose afferent reflex (AAR) and sympathetic activation. This study is to investigate the effects of AAR on lipolysis and the mechanisms of attenuated lipolysis response to enhanced AAR in obesity. Obesity was caused by high-fat diet for 12 weeks in rats. AAR was induced by injection of capsaicin into inguinal WAT or electrical stimulation of epididymal WAT afferent nerve. AAR caused sympathetic activation, which was enhanced in obesity rats. AAR increased cAMP levels and PKA activity, promoted hormone sensitive lipase (HSL) and perilipin phosphorylation, and increased lipolysis in WAT, which were attenuated in obesity rats. PKA activity, cAMP, perilipin and β-adrenoceptor levels were reduced, while HSL was upregulated in adipocytes from obesity rats. In primary adipocytes, isoproterenol increased cAMP levels and PKA activity, promoted HSL and perilipin phosphorylation, and increased lipolysis, which were attenuated in obesity rats. The attenuated effects of isoproterenol in adipocytes from obesity rats were prevented by a cAMP analogue dbcAMP. The results indicate that reduced lipolysis response to enhanced AAR in obesity is attributed to the impaired activation of β-adrenoceptor-cAMP-PKA-HSL pathway. Increased cAMP level in adipocytes rectifies the attenuated lipolysis in obesity.

  3. Triglyceride with medium-chain fatty acids increases the activity and expression of hormone-sensitive lipase in white adipose tissue of C57BL/6J mice.

    Science.gov (United States)

    Liu, Yinghua; Xue, Changyong; Zhang, Yong; Xu, Qing; Yu, Xiaoming; Zhang, Xinsheng; Wang, Jin; Zhang, Rongxin; Gong, Xue; Guo, Changjiang

    2011-01-01

    We have previously shown that medium-chain triglyceride (MCT) resulted in significantly less body fat mass than long-chain triglyceride (LCT) did in hypertriglyceridimic subjects. The possible mechanism for this was investigated by measuring and analyzing changes in the body fat, blood lipid profile, enzymatic level and activity of hormone-sensitive lipase (HSL) and its mRNA expression, and levels of cyclic adenosine monophosphate (cAMP) and protein kinase A (PKA) in white adipose tissue (WAT) of C57BL/6J mice fed for 16 weeks on an MCT or LCT diet. MCT induced lower body weight and body fat, and an improved blood lipid profile than LCT did. The enzymatic level and activity of HSL and its mRNA expression, and the levels of cAMP and PKA were significantly higher in WAT of mice fed with the MCT diet. No significant differences in the levels of lipoprotein lipase and peroxisome proliferator-activated receptor-γ in WAT were apparent between the effects of MCT and LCT. It is concluded that lipolysis by the increased level and activity of HSL, which was induced by the activation of cAMP-dependent PKA in WAT, was partially responsible for the lower fat accumulation in C57BL/6J mice fed with MCT.

  4. Perilipin Promotes HSL-Mediated Adipocyte Lipolysis via Phosphorylation-dependent and Independent Mechanisms

    Science.gov (United States)

    Hormone-sensitive lipase (HSL) is the predominant lipase effector of catecholamine-stimulated lipolysis in adipocytes. HSL-dependent lipolysis, in response to catecholamines, is mediated by protein kinase A (PKA)-dependent phosphorylation of perilipin A (Peri A), an essential lipid droplet (LD)-ass...

  5. Ser649 and Ser650 are the major determinants of protein kinase A-mediated activation of human hormone-sensitive lipase against lipid substrates.

    Directory of Open Access Journals (Sweden)

    Christian Krintel

    Full Text Available BACKGROUND: Hormone-sensitive lipase (HSL is a key enzyme in the mobilization of fatty acids from stored triacylglycerols. Its activity is regulated by reversible protein phosphorylation. In rat HSL Ser563, Ser659 and Ser660 have been shown to be phosphorylated by protein kinase A (PKA in vitro as well as in vivo. METHODOLOGY/PRINCIPAL FINDINGS: In this study we employed site-directed mutagenesis, in vitro phosphorylation and mass spectrometry to show that in vitro phosphorylation of human HSL by PKA occurs primarily on Ser649 and Ser650 (Ser659 and Ser660 in rat HSL. The wild type enzyme and four mutants were expressed in C-terminally His-tagged form in Sf9 insect cells and purified to homogeneity. HSL variants in which Ser552 and/or Ser554 were mutated to Ala or Glu retained both lipolytic and non-lipolytic activity and were phosphorylated by PKA and activated to a similar extent as the wild type enzyme. (32P-labeling studies revealed that the bulk of the phosphorylation was on the Ser649/Ser650 site, with only a minor phosphorylation of Ser552 and Ser554. MS/MS analysis demonstrated that the peptide containing Ser649 and Ser650 was primarily phosphorylated on Ser650. The mutant lacking all four serines had severely reduced lipolytic activity, but a lesser reduction in non-lipolytic activity, had S(0.5 values for p-nitrophenol butyrate and triolein comparable to those of wild type HSL and was not phosphorylated by PKA. PKA phosphorylation of the wild type enzyme resulted in an increase in both the maximum turnover and S(0,5 using the TO substrate. CONCLUSIONS: Our results demonstrate that PKA activates human HSL against lipid substrates in vitro primarily through phosphorylation of Ser649 and Ser650. In addition the results suggest that Ser649 and Ser650 are located in the vicinity of a lipid binding region and that PKA phosphorylation controls the accessibility of this region.

  6. Quantification of hormone sensitive lipase phosphorylation and colocalization with lipid droplets in murine 3T3L1 and human subcutaneous adipocytes via automated digital microscopy and high-content analysis.

    Science.gov (United States)

    McDonough, Patrick M; Ingermanson, Randall S; Loy, Patricia A; Koon, Erick D; Whittaker, Ross; Laris, Casey A; Hilton, Jeffrey M; Nicoll, James B; Buehrer, Benjamin M; Price, Jeffrey H

    2011-06-01

    Lipolysis in adipocytes is associated with phosphorylation of hormone sensitive lipase (HSL) and translocation of HSL to lipid droplets. In this study, adipocytes were cultured in a high-throughput format (96-well dishes), exposed to lipolytic agents, and then fixed and labeled for nuclei, lipid droplets, and HSL (or HSL phosphorylated on serine 660 [pHSLser660]). The cells were imaged via automated digital fluorescence microscopy, and high-content analysis (HCA) methods were used to quantify HSL phosphorylation and the degree to which HSL (or pHSLser660) colocalizes with the lipid droplets. HSL:lipid droplet colocalization was quantified through use of Pearson's correlation, Mander's M1 Colocalization, and the Tanimoto coefficient. For murine 3T3L1 adipocytes, isoproterenol, Lys-γ3-melanocyte stimulating hormone, and forskolin elicited the appearance and colocalization of pHSLser660, whereas atrial natriuretic peptide (ANP) did not. For human subcutaneous adipocytes, isoproterenol, forskolin, and ANP activated HSL phosphorylation/colocalization, but Lys-γ3-melanocyte stimulating hormone had little or no effect. Since ANP activates guanosine 3',5'-cyclic monophosphate (cGMP)-dependent protein kinase, HSL serine 660 is likely a substrate for cGMP-dependent protein kinase in human adipocytes. For both adipocyte model systems, adipocytes with the greatest lipid content displayed the greatest lipolytic responses. The results for pHSLser660 were consistent with release of glycerol by the cells, a well-established assay of lipolysis, and the HCA methods yielded Z' values >0.50. The results illustrate several key differences between human and murine adipocytes and demonstrate advantages of utilizing HCA techniques to study lipolysis in cultured adipocytes.

  7. Involvement of a cGMP-dependent pathway in the natriuretic peptide-mediated hormone-sensitive lipase phosphorylation in human adipocytes.

    Science.gov (United States)

    Sengenes, Coralie; Bouloumie, Anne; Hauner, Hans; Berlan, Michel; Busse, Rudi; Lafontan, Max; Galitzky, Jean

    2003-12-05

    Our previous studies have demonstrated that natriuretic peptides (NPs), peptide hormones with natriuretic, diuretic, and vasodilating properties, exert a potent control on the lipolysis in human adipocytes via the activation of the type A guanylyl cyclase receptor (1, 2). In the current study we investigated the intracellular mechanisms involved in the NP-stimulated lipolytic effect in human preadipocytes and adipocytes. We demonstrate that the atrial NP (ANP)-induced lipolysis in human adipocytes was associated with an enhanced serine phosphorylation of the hormone-sensitive lipase (HSL). Both ANP-mediated lipolysis and HSL phosphorylation were inhibited in the presence of increasing concentrations of the guanylyl cyclase inhibitor LY-83583. ANP did not modulate the activity of the cAMP-dependent protein kinase (PKA). Moreover, H-89, a PKA inhibitor, did not affect the ANP-induced lipolysis. On primary cultures of human preadipocytes, the ANP-mediated lipolytic effect was dependent on the differentiation process. On differentiated human preadipocytes, ANP-mediated lipolysis, associated with an increased phosphorylation of HSL and of perilipin A, was strongly decreased by treatment with the inhibitor of the cGMP-dependent protein kinase I (cGKI), Rp-8-pCPT-cGMPS. Thus, ANP-induced lipolysis in human adipocytes is a cGMP-dependent pathway that induces the phosphorylation of HSL and perilipin A via the activation of cGKI. The present study shows that lipolysis in human adipocytes can be controlled by an independent cGKI-mediated signaling as well as by the classical cAMP/PKA pathway.

  8. Central melanocortin stimulation increases phosphorylated perilipin A and hormone-sensitive lipase in adipose tissues.

    Science.gov (United States)

    Shrestha, Y B; Vaughan, C H; Smith, B J; Song, C K; Baro, D J; Bartness, T J

    2010-07-01

    Norepinephrine (NE) released from the sympathetic nerves innervating white adipose tissue (WAT) is the principal initiator of lipolysis in mammals. Central WAT sympathetic outflow neurons express melanocortin 4-receptor (MC4-R) mRNA. Single central injection of melanotan II (MTII; MC3/4-R agonist) nonuniformly increases WAT NE turnover (NETO), increases interscapular brown adipose tissue (IBAT) NETO, and increases the circulating lipolytic products glycerol and free fatty acid. The WAT pads that contributed to this lipolysis were inferred from the increases in NETO. Because phosphorylation of perilipin A (p-perilipin A) and hormone-sensitive lipase are necessary for NE-triggered lipolysis, we tested whether MTII would increase these intracellular markers of lipolysis. Male Siberian hamsters received a single 3rd ventricular injection of MTII or saline. Trunk blood was collected at 0.5, 1.0, and 2.0 h postinjection from excised inguinal, retroperitoneal, and epididymal WAT (IWAT, RWAT, and EWAT, respectively) and IBAT pads. MTII increased circulating glycerol concentrations at 0.5 and 1.0 h, whereas free fatty acid concentrations were increased at 1.0 and 2.0 h. Western blot analysis showed that MTII specifically increased p-perilipin A and hormone-sensitive lipase only in fat pads that previously had MTII-induced increases in NETO. Phosphorylation increased in IWAT at all time points and IBAT at 0.5 h, but not RWAT or EWAT at any time point. These results show for the first time in rodents that p-perilipin A can serve as an in vivo, fat pad-specific indictor of lipolysis and extend our previous findings showing that central melanocortin stimulation increases WAT lipolysis.

  9. Combined gene and protein expression of hormone-sensitive lipase and adipose triglyceride lipase, mitochondrial content, and adipocyte size in subcutaneous and visceral adipose tissue of morbidly obese men.

    Science.gov (United States)

    De Naeyer, Hélène; Ouwens, D Margriet; Van Nieuwenhove, Yves; Pattyn, Piet; 't Hart, Leen M; Kaufman, Jean-Marc; Sell, Henrike; Eckel, Juergen; Cuvelier, Claude; Taes, Youri E; Ruige, Johannes B

    2011-01-01

    Lipotoxicity in obesity might be a failure of adipocytes to respond sufficiently adequate to persistent energy surplus. To evaluate the role of lipolytic enzymes or mitochondria in lipotoxicity, we studied expression levels of genes and proteins involved in lipolysis and mitochondrial DNA (mtDNA) content. As differences in lipid metabolism between men and women are extremely complex, we recruited only men (lean and morbidly obese) and collected subcutaneous and visceral adipose tissue during abdominal surgery for real-time PCR gene expression, protein expression, and microscopic study. Although mRNA levels of hormone-sensitive lipase (HSL) and adipose triglyceride lipase (ATGL) were increased in visceral adipose tissue of morbidly obese men, this was not paralleled by alterations in protein expression and phosphorylation of HSL and ATGL. mtDNA content of visceral adipose tissue was increased in morbidly obese men as compared to lean controls (p adipocyte size and serum triacylglycerol (r = 0.6, p adipocyte size and CRP (r = 0.6, p dysregulation of involved lipolytic enzymes or adipocyte mitochondria. Copyright © 2011 S. Karger AG, Basel.

  10. Phospholipase C-related catalytically inactive protein (PRIP regulates lipolysis in adipose tissue by modulating the phosphorylation of hormone-sensitive lipase.

    Directory of Open Access Journals (Sweden)

    Toshiya Okumura

    Full Text Available Phosphorylation of hormone-sensitive lipase (HSL and perilipin by protein kinase A (PKA promotes the hydrolysis of lipids in adipocytes. Although activation of lipolysis by PKA has been well studied, inactivation via protein phosphatases is poorly understood. Here, we investigated whether phospholipase C-related catalytically inactive protein (PRIP, a binding partner for protein phosphatase 1 and protein phosphatase 2A (PP2A, is involved in lipolysis by regulating phosphatase activity. PRIP knockout (PRIP-KO mice displayed reduced body-fat mass as compared with wild-type mice fed with standard chow ad libitum. Most other organs appeared normal, suggesting that mutant mice had aberrant fat metabolism in adipocytes. HSL in PRIP-KO adipose tissue was highly phosphorylated compared to that in wild-type mice. Starvation of wild-type mice or stimulation of adipose tissue explants with the catabolic hormone, adrenaline, translocated both PRIP and PP2A from the cytosol to lipid droplets, but the translocation of PP2A was significantly reduced in PRIP-KO adipocytes. Consistently, the phosphatase activity associated with lipid droplet fraction in PRIP-KO adipocytes was significantly reduced and was independent of adrenaline stimulation. Lipolysis activity, as assessed by measurement of non-esterified fatty acids and glycerol, was higher in PRIP-KO adipocytes. When wild-type adipocytes were treated with a phosphatase inhibitor, they showed a high lipolysis activity at the similar level to PRIP-KO adipocytes. Collectively, these results suggest that PRIP promotes the translocation of phosphatases to lipid droplets to trigger the dephosphorylation of HSL and perilipin A, thus reducing PKA-mediated lipolysis.

  11. Adipose triglyceride lipase and hormone-sensitive lipase are involved in fat loss in JunB-deficient mice.

    Science.gov (United States)

    Pinent, Montserrat; Prokesch, Andreas; Hackl, Hubert; Voshol, Peter J; Klatzer, Ariane; Walenta, Evelyn; Panzenboeck, Ute; Kenner, Lukas; Trajanoski, Zlatko; Hoefler, Gerald; Bogner-Strauss, Juliane G

    2011-07-01

    Proteins of the activator protein-1 family are known to have roles in many physiological processes such as proliferation, apoptosis, and inflammation. However, their role in fat metabolism has yet to be defined in more detail. Here we study the impact of JunB deficiency on the metabolic state of mice. JunB knockout (JunB-KO) mice show markedly decreased weight gain, reduced fat mass, and a low survival rate compared with control mice. If fed a high-fat diet, the weight gain of JunB-KO mice is comparable to control mice and the survival rate improves dramatically. Along with normal expression of adipogenic marker genes in white adipose tissue (WAT) of JunB-KO mice, this suggests that adipogenesis per se is not affected by JunB deficiency. This is supported by in vitro data, because neither JunB-silenced 3T3-L1 cells nor mouse embryonic fibroblasts from JunB-KO mice show a change in adipogenic potential. Interestingly, the key enzymes of lipolysis, adipose triglyceride lipase and hormone-sensitive lipase, were significantly increased in WAT of fasted JunB-KO mice. Concomitantly, the ratio of plasma free fatty acids per gram fat mass was increased, suggesting an elevated lipolytic rate under fasting conditions. Furthermore, up-regulation of TNFα and reduced expression of perilipin indicate that this pathway is also involved in increased lipolytic rate in these mice. Additionally, JunB-KO mice are more insulin sensitive than controls and show up-regulation of lipogenic genes in skeletal muscle, indicating a shuttling of energy substrates from WAT to skeletal muscle. In summary, this study provides valuable insights into the impact of JunB deficiency on the metabolic state of mice.

  12. Fatty Acid Synthase and Hormone-sensitive Lipase Expression in Liver Are Involved in Zinc-α2-glycoprotein-induced Body Fat Loss in Obese Mice

    Institute of Scientific and Technical Information of China (English)

    Feng-ying Gong; Jie-ying Deng; Hui-juan Zhu; Hui Pan; Lin-jie Wang; Hong-bo Yang

    2010-01-01

    Objective To explore the effects of zinc-a2-glycoprotein (ZAG) on body weight and body fat in high-fat-diet (HFD)-induced obesity in mice and the possible mechanism.Methods Thirty-six male mice were fed with standard food (SF) (n=9) and HFD (n=27), respec-tively. Five weeks later, 9 mice fed with HFD were subjected to ZAG expression plasmid DNA transfection by liposome transfection method, and another 9 mice to negative control plasmid transfection. Two weeks later, serum ZAG level in the mice was assayed by Western blot, and the effects of ZAG over-expression on body weight, body fat, serum biochemical indexes, and adipose tissue of obese mice were evaluated. The mRNA expressions of fatty acid synthase (FAS) and hormone-sensitive lipase (HSL) in liver tissue were de-termined by reverse transcription-polymerase chain reaction.Results Serum ZAG level significantly lowered in simple HFD-fed mice in comparison to SF-fed mice (0.51±0.10 AU vs. 0.75±0.07 AU, P<0.01). Further statistical analysis demonstrated that ZAG level was negatively correlated with body weight (r =-0.56, P<0.001), epididymal fat mass (r=-0. 67, P<0.001), percentage of epididymal fat (r=-0.65, P<0.001 ), and increased weight (r=-0.57, P<0.001) in simple SF-and HFD-fed mice. ZAG over-expression in obese mice reduced body weight and the percentage of epididy-mal fat. Furthermore, FAS mRNA expression decreased (P<0.01) and HSL mRNA expression increased (P<0.001) in the liver in ZAG over-expressing mice.Conclusions ZAG is closely related to obesity. Serum ZAG level is inversely correlated with body weight and percentage of body fat. The action of ZAG is associated with reduced FAS expression and in-creased HSL expression in the liver of obese mice.

  13. Activity and tissue-specific expression of lipases and tumor-necrosis factor alpha in lean and obese cats.

    NARCIS (Netherlands)

    Hoenig, M.; McGoldrick, J.B.; Beer, M. de; Demacker, P.N.M.; Ferguson, D.C.

    2006-01-01

    Post-heparin plasma activity of lipoprotein lipase (LPL) and hepatic lipase (HL), and fat and muscle activity of LPL were measured in neutered lean and obese cats. Lipoprotein lipase, hormone-sensitive lipase (HSL), and tumor necrosis factor a (TNF) mRNA were measured in muscle and fat tissue with r

  14. Activity and tissue-specific expression of lipases and tumor-necrosis factor alpha in lean and obese cats.

    NARCIS (Netherlands)

    Hoenig, M.; McGoldrick, J.B.; Beer, M. de; Demacker, P.N.M.; Ferguson, D.C.

    2006-01-01

    Post-heparin plasma activity of lipoprotein lipase (LPL) and hepatic lipase (HL), and fat and muscle activity of LPL were measured in neutered lean and obese cats. Lipoprotein lipase, hormone-sensitive lipase (HSL), and tumor necrosis factor a (TNF) mRNA were measured in muscle and fat tissue with r

  15. Activity and tissue-specific expression of lipases and tumor-necrosis factor alpha in lean and obese cats.

    NARCIS (Netherlands)

    Hoenig, M.; McGoldrick, J.B.; Beer, M. de; Demacker, P.N.M.; Ferguson, D.C.

    2006-01-01

    Post-heparin plasma activity of lipoprotein lipase (LPL) and hepatic lipase (HL), and fat and muscle activity of LPL were measured in neutered lean and obese cats. Lipoprotein lipase, hormone-sensitive lipase (HSL), and tumor necrosis factor a (TNF) mRNA were measured in muscle and fat tissue with

  16. Berberine increases adipose triglyceride lipase in 3T3-L1 adipocytes through the AMPK pathway

    OpenAIRE

    Jiang, Dongqing; Wang, Dianhui; ZHUANG, XIANGHUA; Wang, Zhanqing; Ni, Yihong; Chen, Shihong; Sun, Fudun

    2016-01-01

    Background Obesity is closely related to the metabolism of triacylglycerol (TG) in adipocytes. Adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (HSL) are rate-limiting enzymes that control the hydrolysis of TG. Effects on ATGL and HSL to increase lipolysis may counteract obesity. Berberine (BBR) is a compound derived from the Chinese medicine plant Coptis chinensis. In the present study we show the effects of BBR on ATGL and HSL and explore the potential underlying mechanisms o...

  17. Purification, crystallization and preliminary crystallographic analysis of Est25: a ketoprofen-specific hormone-sensitive lipase

    Energy Technology Data Exchange (ETDEWEB)

    Kim, SeungBum [Department of Biological and Molecular Engineering, College of Engineering, Ajou University, Suwon (Korea, Republic of); Joo, Sangbum [Department of Molecular Cell Biology, Sungkyunkwan University School of Medicine, Suwon (Korea, Republic of); Yoon, Hyun C.; Ryu, Yeonwoo [Department of Biological and Molecular Engineering, College of Engineering, Ajou University, Suwon (Korea, Republic of); Kim, Kyeong Kyu, E-mail: kkim@med.skku.ac.kr [Department of Molecular Cell Biology, Sungkyunkwan University School of Medicine, Suwon (Korea, Republic of); Kim, T. Doohun, E-mail: kkim@med.skku.ac.kr [Department of Biological and Molecular Engineering, College of Engineering, Ajou University, Suwon (Korea, Republic of)

    2007-07-01

    Est25, a ketoprofen-specific hormone-sensitive lipase from a metagenomic library, was crystallized and diffraction data were collected to 1.49 Å resolution. Ketoprofen, a nonsteroidal anti-inflammatory drug, inhibits the synthesis of prostaglandin. A novel hydrolase (Est25) with high ketoprofen specificity has previously been identified using a metagenomic library from environmental samples. Recombinant Est25 protein with a histidine tag at the N-terminus was expressed in Escherichia coli and purified in a homogenous form. Est25 was crystallized from 2.4 M sodium malonate pH 7.0 and X-ray diffraction data were collected to 1.49 Å using synchrotron radiation. The crystals belong to the monoclinic space group C2, with unit-cell parameters a = 197.8, b = 95.2, c = 99.4 Å, β = 97.1°.

  18. Study on the Polymorphism of Porcine Hormone-sensitive Lipase Gene Exon I by PCR-RFLP in Different Breeds%猪品种间HSL基因外显子I PCR-RFLP的研究

    Institute of Scientific and Technical Information of China (English)

    雷明刚; 吴珍芳; 邓昌彦; 熊远著

    2001-01-01

    Hormone-sensitive lipase (HSL) is the key enzyme responsible for the mobilization of free acids from adipose tissue, and it is also the most important enzyme that affects fat deposition. In this paper, the polymorphism of HSL gene in exon I was detected by PCR-RFLP. It revealed a BsaHI-site polymorphism in different pig breeds. There exist two genotypes (AA,AG) in the fat-type Tongcheng pig and Qingping pig with allele frequencies of 0.917A,0.083G and 0.893,0.107,respectively. Duroc has two genotypes (AG,GG) with allele frequencies 0.15A and 0.85G. However, There exists only GG genotype in lean-type Large White and Landrace pig.%激素敏感脂肪酶(Hormone-sensitive lipase,HSL)是负责分解脂肪组织中甘油三酯释放游离脂肪酸的关键酶。本研究利用PCR*.RFLP(polymerase chain reaction-restriction fragment length polymorphism)方法,分析了猪HSL基因外显子I区多态性,发现不同品种猪间存在多态性。瘦肉型大白猪和长白猪全部表现为GG基因型;杜洛克猪表现为AG和GG两种基因型,其等位基因A和G的频率是15%和85%;脂肪型通城猪和清平猪表现为AA和AG两种基因型,其等位基因A、G的频率分别是91.67%、8.33%和89.83%、10.17%。

  19. Agonists of the G protein-coupled receptor 109A-mediated pathway promote antilipolysis by reducing serine residue 563 phosphorylation of hormone-sensitive lipase in bovine adipose tissue explants.

    Science.gov (United States)

    Kenéz, A; Locher, L; Rehage, J; Dänicke, S; Huber, K

    2014-01-01

    A balanced lipolytic regulation in adipose tissues based on fine-tuning of prolipolytic and antilipolytic pathways is of vital importance to maintain the metabolic health in dairy cows. Antilipolytic pathways, such as the G protein-coupled receptor 109A (GPR109A)-mediated pathway and the insulin signaling pathway in bovine adipose tissues may be involved in prohibiting excessive lipomobilization by reducing triglycerol hydrolysis. This study aimed to evaluate the in vitro antilipolytic potential of the mentioned pathways in bovine adipose tissue explants. Therefore, subcutaneous and retroperitoneal adipose tissue samples (approximately 100mg) of German Holstein cows were treated for 90 min ex vivo with nicotinic acid (2, 8, or 32 μM), nicotinamide (2, 8, or 32 μM), β-hydroxybutyrate (0.2, 1, or 5mM), or insulin (12 mU/L), with a concurrent lipolytic challenge provoked with 1 μM isoproterenol. Lipolytic and antilipolytic responses of the adipose tissues were assessed by measuring free glycerol and nonesterified fatty acid release. To identify molecular components of the investigated antilipolytic pathways, protein abundance of GPR109A and the extent of hormone-sensitive lipase (HSL) phosphorylation at serine residue 563 were detected by Western blotting. Treatment with nicotinic acid or β-hydroxybutyrate decreased the lipolytic response in adipose tissue explants and concurrently reduced the extent of HSL phosphorylation, but treatment with nicotinamide or insulin did not. Subcutaneous adipose tissue constitutively expressed more GPR109A protein, but no other depot-specific differences were observed. This study provides evidence that the GPR109A-mediated pathway is functionally existent in bovine adipose tissues, and confirms that HSL phosphorylation at serine residue 563 is also important in antilipolytic regulation in vitro. This antilipolytic pathway may be involved in a balanced lipid mobilization in the dairy cow.

  20. Absence of cardiac lipid accumulation in transgenic mice with heart-specific HSL overexpression.

    Science.gov (United States)

    Suzuki, J; Shen, W J; Nelson, B D; Patel, S; Veerkamp, J H; Selwood, S P; Murphy, G M; Reaven, E; Kraemer, F B

    2001-10-01

    Hormone-sensitive lipase (HSL) hydrolyzes triglyceride (TG) in adipose tissue. HSL is also expressed in heart. To explore the actions of cardiac HSL, heart-specific, tetracycline (Tc)-controlled HSL-overexpressing mice were generated. Tc-responsive element-HSL transgenic (Tg) mice were generated and crossed with myosin heavy chain (MHC)alpha-tTA Tg mice, which express the Tc-responsive transactivator (tTA) in the heart. The double-Tg mice (MHC-HSL) were maintained with doxycycline (Dox) to suppress Tg HSL. Upon removal of Dox, cardiac HSL activity and protein increased 12- and 8-fold, respectively, and the expression was heart specific. Although cardiac TG content increased twofold in control mice after an overnight fast, it did not increase in HSL-induced mice. Electron microscopy showed numerous lipid droplets in the myocardium of fasted control mice, whereas fasted HSL-induced mice showed virtually no droplets. Microarray analysis showed altered expression of cardiac genes for fatty acid oxidation, transcription factors, signaling molecules, cytoskeletal proteins, and histocompatibility antigens in HSL-induced mice. Thus cardiac HSL plays a role in controlling accumulation of triglyceride droplets and can affect the expression of a number of cardiac genes.

  1. Identification of peroxisome-proliferator responsive element in the mouse HSL gene.

    Science.gov (United States)

    Yajima, Hiroaki; Kobayashi, Yumie; Kanaya, Tomoka; Horino, Yoko

    2007-01-12

    Hormone-sensitive lipase (HSL) catalyzes the rate-limiting step of lipolysis in adipose tissue. Several studies suggest that protein phosphorylation regulates the HSL enzymatic activity. On the other hand, the precise mechanism of the transcriptional regulation of the HSL gene remains to be elucidated. Here, we identified a functional peroxisome-proliferator responsive element (PPRE) in the mouse HSL promoter by reporter assay in CV-1 cells using serial deletion and point mutants of the 5'-flanking region. Chromatin immunoprecipitation (ChIP) analysis revealed that both peroxisome-proliferator activated receptor (PPARgamma) and retinoid X receptor (RXRalpha) interacted with the region. Binding of the PPARgamma/RXRalpha heterodimer to the PPRE sequence was also confirmed by electrophoretic mobility shift assay. These results indicate that the HSL gene is transcriptionally regulated by PPARgamma/RXRalpha heterodimer, and suggest that a cis-acting element regulates the HSL gene expression.

  2. Lipolysis and lipases in white adipose tissue – An update

    OpenAIRE

    Bolsoni-Lopes,Andressa; Alonso-Vale, Maria Isabel C.

    2015-01-01

    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 mechanism...

  3. Adequate evaluation of HSL mass and activity in rat adipose tissue in fasting and aging-related obesity.

    Science.gov (United States)

    Tsujita, Takahiro; Sumiyoshi, Maho; Morimoto, Chie; Kameda, Kenji; Okuda, Hiromichi

    2002-04-01

    Adipose tissue is a unique tissue because its mass is readily changed by altering nutritional conditions. Therefore the activity and content of enzyme in the adipose tissue is significantly differed according to the way of their presentation: per g tissue, per whole tissue, or per cell number. In the present study, the effects of the ways of expressing the hormone sensitive lipase (HSL) activity and content were studied in rat by decreasing or increasing adipose tissue. Fasting caused a progressive decline in body weight and in the weight of the epididymal fat pad. When the HSL content was expressed per g of adipose tissue, the lipase activity and immunoreactive HSL protein content in fasting rats were higher than those in fed rats. On the other hand, when they were expressed as per fat pad, the lipase activity and immunoreactive HSL protein in fasting rats were lower than those in fed rats. The opposite results were observed in obesity. When the HSL content was expressed per g of adipose tissue, the lipase activity and immunoreactive HSL protein in obese rats were lower than in control rats. However, when the HSL content was expressed per fat pad, the lipase activity and immunoreactive HSL protein in the obese rats were higher than in the control rats. Therefore we must pay careful attention to the way of presentation of adipose tissue enzyme contents.

  4. Regulation of HSL serine phosphorylation in skeletal muscle and adipose tissue.

    Science.gov (United States)

    Watt, Matthew J; Holmes, Anna G; Pinnamaneni, Srijan K; Garnham, Andrew P; Steinberg, Gregory R; Kemp, Bruce E; Febbraio, Mark A

    2006-03-01

    Hormone-sensitive lipase (HSL) is important for the degradation of triacylglycerol in adipose and muscle tissue, but the tissue-specific regulation of this enzyme is not fully understood. We investigated the effects of adrenergic stimulation and AMPK activation in vitro and in circumstances where AMPK activity and catecholamines are physiologically elevated in humans in vivo (during physical exercise) on HSL activity and phosphorylation at Ser(563) and Ser(660), the PKA regulatory sites, and Ser(565), the AMPK regulatory site. In human experiments, skeletal muscle, subcutaneous adipose and venous blood samples were obtained before, at 15 and 90 min during, and 120 min after exercise. Skeletal muscle HSL activity was increased by approximately 80% at 15 min compared with rest and returned to resting rates at the cessation of and 120 min after exercise. Consistent with changes in plasma epinephrine, skeletal muscle HSL Ser(563) and Ser(660) phosphorylation were increased by 27% at 15 min (P HSL Ser(565) phosphorylation and AMPK signaling were increased at 90 min during, and after, exercise. Phosphorylation of adipose tissue HSL paralleled changes in skeletal muscle in vivo, except HSL Ser(660) was elevated 80% in adipose compared with 35% in skeletal muscle during exercise. Studies in L6 myotubes and 3T3-L1 adipocytes revealed important tissue differences in the regulation of HSL. AMPK inhibited epinephrine-induced HSL activity in L6 myotubes and was associated with reduced HSL Ser(660) but not Ser(563) phosphorylation. HSL activity was reduced in L6 myotubes expressing constitutively active AMPK, confirming the inhibitory effects of AMPK on HSL activity. Conversely, in 3T3-L1 adipocytes, AMPK activation after epinephrine stimulation did not prevent HSL activity or glycerol release, which coincided with maintenance of HSL Ser(660) phosphorylation. Taken together, these data indicate that HSL activity is maintained in the face of AMPK activation as a result of

  5. Structural and kinetic overview of the carboxylesterase EST2 from alicyclobacillus acidocaldarius: a comparison with the other members of the HSL family.

    Science.gov (United States)

    Mandrich, Luigi; Merone, Luigia; Manco, Giuseppe

    2009-01-01

    Thermophilic and hyperthermophilic carboxylesterases (EC 3.1.1.1) are excellent model systems for studying structure function relationships as well as in vitro and in vivo evolution and possible biotechnological applications. In this paper we review the main aspect of one of most studied microbial representative of the hormone sensitive lipase family (HSL), namely carboxylesterase 2 (EST2) from Alicyclobacillus acidocaldarius.

  6. Cholesteryl ester hydrolase activity is abolished in HSL-/- macrophages but unchanged in macrophages lacking KIAA1363.

    Science.gov (United States)

    Buchebner, Marlene; Pfeifer, Thomas; Rathke, Nora; Chandak, Prakash G; Lass, Achim; Schreiber, Renate; Kratzer, Adelheid; Zimmermann, Robert; Sattler, Wolfgang; Koefeler, Harald; Fröhlich, Eleonore; Kostner, Gerhard M; Birner-Gruenberger, Ruth; Chiang, Kyle P; Haemmerle, Guenter; Zechner, Rudolf; Levak-Frank, Sanja; Cravatt, Benjamin; Kratky, Dagmar

    2010-10-01

    Cholesteryl ester (CE) accumulation in macrophages represents a crucial event during foam cell formation, a hallmark of atherogenesis. Here we investigated the role of two previously described CE hydrolases, hormone-sensitive lipase (HSL) and KIAA1363, in macrophage CE hydrolysis. HSL and KIAA1363 exhibited marked differences in their abilities to hydrolyze CE, triacylglycerol (TG), diacylglycerol (DG), and 2-acetyl monoalkylglycerol ether (AcMAGE), a precursor for biosynthesis of platelet-activating factor (PAF). HSL efficiently cleaved all four substrates, whereas KIAA1363 hydrolyzed only AcMAGE. This contradicts previous studies suggesting that KIAA1363 is a neutral CE hydrolase. Macrophages of KIAA1363(-/-) and wild-type mice exhibited identical neutral CE hydrolase activity, which was almost abolished in tissues and macrophages of HSL(-/-) mice. Conversely, AcMAGE hydrolase activity was diminished in macrophages and some tissues of KIAA1363(-/-) but unchanged in HSL(-/-) mice. CE turnover was unaffected in macrophages lacking KIAA1363 and HSL, whereas cAMP-dependent cholesterol efflux was influenced by HSL but not by KIAA1363. Despite decreased CE hydrolase activities, HSL(-/-) macrophages exhibited CE accumulation similar to wild-type (WT) macrophages. We conclude that additional enzymes must exist that cooperate with HSL to regulate CE levels in macrophages. KIAA1363 affects AcMAGE hydrolase activity but is of minor importance as a direct CE hydrolase in macrophages.

  7. Differential effects of high MUFA with high or low P/S ratio (polyunsaturated to saturated fatty acids) on improving hepatic lipolytic enzymes and mediating PPARγ related with lipoprotein lipase and hormone-sensitive lipase of white adipose tissue in diet-induced obese hamster.

    Science.gov (United States)

    Liao, F-H; Liou, T-H; Chiu, W-C; Shieh, M-J; Chien, Y-W

    2010-11-01

    The aim of this study was to assess the relationship between high monounsaturated fatty acids (MUFAs) with different levels of polyunsaturated-to-saturated fatty acid (P/S) ratios and body fat loss in diet-induced obesity (DIO) models. Male Golden Syrian hamsters were randomly assigned to the control group (n=12) and obesity group (n=24) for 4 weeks of the high-fat DIO period; afterward, six hamsters from each group were killed. The remaining control hamsters were still fed a low-fat diet. For an additional 8 weeks, the remaining obesity hamsters were switched to a low-fat diet and subdivided into three subgroups (n=6/group): the obesity-control (ObC) group, high MUFA with high P/S ratio oil (HMHR) group and olive oil (OO) group. Serum insulin and leptin concentrations were measured, and hepatic fatty acid metabolic enzymes and adipose differentiation markers were determined using enzyme activities analysis, western blot and semiquantification reverse-transcription PCR. No difference was observed in the mean energy intake through all study periods. After the DIO period, the obesity group increased in weight gain and epididymal fat weight compared with the control group. DIO hamsters in the HMLR group had significant reductions in white adipose tissue deposition and plasma leptin levels, suppression in adipose peroxisome proliferator-activated receptor-γ (PPARγ) and lipoprotein lipase (LPL) mRNA expressions and increases in hepatic acyl-CoA oxidase and carnitine palmitoyltransferase-I activities and mRNA levels compared with those in the ObC group. The HMHR group had upregulated phosphorylation of hormone-sensitive lipase (HSL) relative to total HSL protein levels compared with the OO group. However, the OO group had significantly elevated hepatic de novo lipogenesis compared with the HMHR group. HMHR seemed to be beneficial in depleting white adipose tissue accumulation by decreasing adipose PPARγ and LPL mRNA expressions and mediating phosphorylation of HSL

  8. Adipocyte lipases and defect of lipolysis in human obesity.

    Science.gov (United States)

    Langin, Dominique; Dicker, Andrea; Tavernier, Geneviève; Hoffstedt, Johan; Mairal, Aline; Rydén, Mikael; Arner, Erik; Sicard, Audrey; Jenkins, Christopher M; Viguerie, Nathalie; van Harmelen, Vanessa; Gross, Richard W; Holm, Cecilia; Arner, Peter

    2005-11-01

    The mobilization of fat stored in adipose tissue is mediated by hormone-sensitive lipase (HSL) and the recently characterized adipose triglyceride lipase (ATGL), yet their relative importance in lipolysis is unknown. We show that a novel potent inhibitor of HSL does not inhibit other lipases. The compound counteracted catecholamine-stimulated lipolysis in mouse adipocytes and had no effect on residual triglyceride hydrolysis and lipolysis in HSL-null mice. In human adipocytes, catecholamine- and natriuretic peptide-induced lipolysis were completely blunted by the HSL inhibitor. When fat cells were not stimulated, glycerol but not fatty acid release was inhibited. HSL and ATGL mRNA levels increased concomitantly during adipocyte differentiation. Abundance of the two transcripts in human adipose tissue was highly correlated in habitual dietary conditions and during a hypocaloric diet, suggesting common regulatory mechanisms for the two genes. Comparison of obese and nonobese subjects showed that obesity was associated with a decrease in catecholamine-induced lipolysis and HSL expression in mature fat cells and in differentiated preadipocytes. In conclusion, HSL is the major lipase for catecholamine- and natriuretic peptide-stimulated lipolysis, whereas ATGL mediates the hydrolysis of triglycerides during basal lipolysis. Decreased catecholamine-induced lipolysis and low HSL expression constitute a possibly primary defect in obesity.

  9. ATGL and HSL are not coordinately regulated in response to fuel partitioning in fasted rats.

    Science.gov (United States)

    Bertile, Fabrice; Raclot, Thierry

    2011-04-01

    Prolonged fasting is characterized by lipid mobilization (Phase 2), followed by protein breakdown (Phase 3). Knowing that body lipids are not exhausted in Phase 3, we investigated whether changes in the metabolic status of prolonged fasted rats are associated with differences in the expression of epididymal adipose tissue proteins involved in lipid mobilization. The final body mass, body lipid content, locomotor activity and metabolite and hormone plasma levels differed between groups. Compared with fed rats, adiposity and epididymal fat mass decreased in Phase 2 (approximately two- to threefold) and Phase 3 (∼4.5-14-fold). Plasma nonesterified fatty acids (NEFA) concentrations were increased in Phase 2 (approximately twofold) and decreased in Phase 3 (approximately twofold). Daily locomotor activity was markedly increased in Phase 3 (∼11-fold). Compared with the fed state, expressions of adipose triglyceride lipase (ATGL; mRNA and protein), hormone-sensitive lipase (HSL; mRNA) and phosphorylated HSL at residue Ser660 (HSL Ser(660)) were increased during Phase 2 (∼1.5-2-fold). HSL (mRNA and protein) and HSL Ser(660) levels were lowered during Phase 3 (∼3-12-fold). Unlike HSL and HSL Ser(660), ATGL expression did not correlate with circulating NEFA, mostly due to data from animals in Phase 3. At this stage, ATGL could play an essential role for maintaining a low mobilization rate of NEFA, possibly to sustain muscle performance and hence increased locomotor activity. We conclude that ATGL and HSL are not coordinately regulated in response to changes in fuel partitioning during prolonged food deprivation, ATGL appearing as the major lipase in late fasting.

  10. Progesterone-induced down-regulation of hormone sensitive lipase (Lipe) and up-regulation of G0/G1 switch 2 (G0s2) genes expression in inguinal adipose tissue of female rats is reflected by diminished rate of lipolysis.

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    Stelmanska, Ewa; Szrok, Sylwia; Swierczynski, Julian

    2015-03-01

    Decreased lipolytic activity in adipose tissue may be one of the reasons behind excess accumulation of body fat during pregnancy. The aim of this study was to analyze the effect of progesterone on the expression of: (a) Lipe (encoding hormone-sensitive lipase, HSL), (b) Pnpla2 (encoding adipose triglyceride lipase, ATGL), (c) abhydrolase domain containing 5 (Abhd5), and (d) G0/G1 switch 2 (G0s2) genes in white adipose tissue (WAT), as potential targets for progesterone action during the course of pregnancy. Administration of progesterone to female rats, which was reflected by approximately 2.5-fold increase in circulating progesterone concentration, is associated with a decrease in Lipe gene expression in the inguinal WAT. The expression of Pnpla2 gene in all main fat depots of females and males remained unchanged after progesterone administration. Administration of progesterone resulted in an increase in the expression of Abhd5 gene (whose product increases ATGL activity) and G0s2 gene (whose product decreases ATGL activity) in the inguinal WAT of female rats. Mifepristone, a selective antagonist of progesterone receptor, abolished the effect of progesterone on Lipe, Abhd5 and G0s2 genes expression in the inguinal WAT. The decrease in Lipe and the increase in Abhd5 and G0s2 genes expression was associated with lower rate of stimulated lipolysis. Administration of progesterone exerted no effect on Lipe, Abhd5 and G0s2 genes expression and stimulated lipolysis in the retroperitoneal WAT of females, as well as in the inguinal, epididymal and retroperitoneal WAT of males. In conclusion, our findings suggest that progesterone decreases the rate of lipolysis in the inguinal WAT of female rats, inhibiting the activity of both ATGL (by stimulating synthesis of G0S2 - specific inhibitor of the enzyme) and HSL (due to inhibition of Lipe gene expression). Copyright © 2014. Published by Elsevier Ltd.

  11. Human adipose triglyceride lipase (PNPLA2) is not regulated by obesity and exhibits low in vitro triglyceride hydrolase activity.

    Science.gov (United States)

    Mairal, A; Langin, D; Arner, P; Hoffstedt, J

    2006-07-01

    The recent identification of murine adipose triglyceride lipase (ATGL, now known as patatin-like phospholipase domain containing 2 [PNPLA2]), gene product of Pnpla2, has questioned the unique role of hormone sensitive lipase (HSL, now known as LIPE), gene product of Lipe, in fat cell lipolysis. Here, we investigated human ATGL and HSL adipose tissue gene expression and in vitro lipase activity. Levels of mRNA in adipose tissue from healthy obese and non-obese subjects were measured and lipase activity and adipocyte lipolytic capacity determined. HSL and ATGL cDNAs were transfected into Cos-7 cells and the relative tri- and diglyceride hydrolase activities were measured. Obesity was associated with a decreased subcutaneous and increased omental adipose tissue level of HSL mRNA. Subcutaneous HSL mRNA content was normalised upon weight reduction. In contrast, ATGL mRNA levels were unaffected by obesity and weight reduction. A high adipose tissue lipase activity was associated with increased maximal lipolysis and increased HSL, but not with ATGL mRNA levels. The in vitro triglyceride hydrolase activity of HSL was markedly higher than that of ATGL and contrary to HSL, ATGL was devoid of diglyceride hydrolase activity. The use of a selective HSL-inhibitor resulted in complete inhibition of HSL-mediated tri- and diglyceride hydrolase activity. The pH profile of human white adipose tissue triolein hydrolase activity was identical to that of HSL but differed from the ATGL profile. HSL, but not ATGL gene expression shows a regulation according to obesity status and is associated with increased adipose tissue lipase activity. Moreover, HSL has a higher capacity than ATGL to hydrolyse triglycerides in vitro.

  12. HSL-knockout mouse testis exhibits class B scavenger receptor upregulation and disrupted lipid raft microdomains.

    Science.gov (United States)

    Casado, María Emilia; Huerta, Lydia; Ortiz, Ana Isabel; Pérez-Crespo, Mirian; Gutiérrez-Adán, Alfonso; Kraemer, Fredric B; Lasunción, Miguel Ángel; Busto, Rebeca; Martín-Hidalgo, Antonia

    2012-12-01

    There is a tight relationship between fertility and changes in cholesterol metabolism during spermatogenesis. In the testis, class B scavenger receptors (SR-B) SR-BI, SR-BII, and LIMP II mediate the selective uptake of cholesterol esters from HDL, which are hydrolyzed to unesterified cholesterol by hormone-sensitive lipase (HSL). HSL is critical because HSL knockout (KO) male mice are sterile. The aim of the present work was to determine the effects of the lack of HSL in testis on the expression of SR-B, lipid raft composition, and related cell signaling pathways. HSL-KO mouse testis presented altered spermatogenesis associated with decreased sperm counts, sperm motility, and infertility. In wild-type (WT) testis, HSL is expressed in elongated spermatids; SR-BI, in Leydig cells and spermatids; SR-BII, in spermatocytes and spermatids but not in Leydig cells; and LIMP II, in Sertoli and Leydig cells. HSL knockout male mice have increased expression of class B scavenger receptors, disrupted caveolin-1 localization in lipid raft plasma membrane microdomains, and activated phospho-ERK, phospho-AKT, and phospho-SRC in the testis, suggesting that class B scavenger receptors are involved in cholesterol ester uptake for steroidogenesis and spermatogenesis in the testis.

  13. The crystal structure of an HSL-homolog EstE5 complex with PMSF reveals a unique configuration that inhibits the nucleophile Ser144 in catalytic triads.

    Science.gov (United States)

    Nam, Ki Hyun; Kim, Soo-Jin; Priyadarshi, Amit; Kim, Hyun Sook; Hwang, Kwang Yeon

    2009-11-13

    The esterase/lipase family (EC 3.1.1.3/EC 3.1.1.1) represents a diverse group of hydrolases that catalyze the cleavage of ester bonds and are widely distributed in animals, plants and microorganisms. Among these enzymes, hormone-sensitive lipases, play a critical role in the regulation of rodent fat cell lipolysis and are regarded as adipose tissue-specific enzymes. Recently, we reported the structural and biological characterization of EstE5 from the metagenome library [K.H. Nam, M.Y. Kim, S.J. Kim, A. Priyadarshi, W.H. Lee, K.Y. Hwang, Structural and functional analysis of a novel EstE5 belonging to the subfamily of hormone-sensitive lipase, Biochem. Biophys. Res. Commun. 379 (2009) 553-556]. The structure of this protein revealed that it belongs to the HSL-family. Here, we report the inhibition of the activity of the HSL-homolog EstE5 protein as determined by the use of esterase/lipase inhibitors. Our results revealed that the EstE5 protein is significantly inhibited by PMSF. In addition, this is the first study to identify the crystal structures of EstE5-PMSF at 2.4 and 2.5A among the HSL-homolog structures. This structural configuration is similar to that adopted when serine proteases are inhibited by PMSF. The results presented here provide valuable information regarding the properties of the HSL-family.

  14. Masoprocol decreases rat lipolytic activity by decreasing the phosphorylation of HSL.

    Science.gov (United States)

    Gowri, M S; Azhar, R K; Kraemer, F B; Reaven, G M; Azhar, S

    2000-09-01

    Masoprocol (nordihydroguaiaretic acid), a lipoxygenase inhibitor isolated from the creosote bush, has been shown to decrease adipose tissue lipolytic activity both in vivo and in vitro. The present study was initiated to test the hypothesis that the decrease in lipolytic activity by masoprocol resulted from modulation of adipose tissue hormone-sensitive lipase (HSL) activity. The results indicate that oral administration of masoprocol to rats with fructose-induced hypertriglyceridemia significantly decreased their serum free fatty acid (FFA; P HSL activity were significantly lower (P HSL protein. Incubation of masoprocol with adipocytes from chow-fed rats significantly inhibited isoproterenol-induced lipolytic activity and HSL activity, associated with a decrease in the ability of isoproterenol to phosphorylate HSL. Masoprocol had no apparent effect on adipose tissue phosphatidylinositol 3-kinase activity, but okadaic acid, a serine/threonine phosphatase inhibitor, blocked the antilipolytic effect of masoprocol. The results of these in vitro and in vivo experiments suggest that the antilipolytic activity of masoprocol is secondary to its ability to inhibit HSL phosphorylation, possibly by increasing phosphatase activity. As a consequence, masoprocol administration results in lower serum FFA and TG concentrations in hypertriglyceridemic rodents.

  15. Adrenergic regulation of HSL serine phosphorylation and activity in human skeletal muscle during the onset of exercise.

    Science.gov (United States)

    Talanian, Jason L; Tunstall, Rebecca J; Watt, Matthew J; Duong, Mylinh; Perry, Christopher G R; Steinberg, Gregory R; Kemp, Bruce E; Heigenhauser, George J F; Spriet, Lawrence L

    2006-10-01

    Skeletal muscle hormone-sensitive lipase (HSL) activity is increased by contractions and increases in blood epinephrine (EPI) concentrations and cyclic AMP activation of the adrenergic pathway during prolonged exercise. To determine the importance of hormonal stimulation of HSL activity during the onset of moderate- and high-intensity exercise, nine men [age 24.3 +/- 1.2 yr, 80.8 +/- 5.0 kg, peak oxygen consumption (VO2 peak) 43.9 +/- 3.6 ml x kg(-1) x min(-1)] cycled for 1 min at approximately 65% VO2 peak, rested for 60 min, and cycled at approximately 90% VO2 peak for 1 min. Skeletal muscle biopsies were taken pre- and postexercise, and arterial blood was sampled throughout exercise. Arterial EPI increased (P HSL activity increased (P HSL Ser660 phosphorylation (approximately 55% increase) and ERK1/2 phosphorylation ( approximately 33% increase) were augmented following exercise at both intensities, whereas HSL Ser563 and Ser565 phosphorylation were not different from rest. The results indicate that increases in arterial EPI concentration during the onset of moderate- and high-intensity exercise increase cyclic AMP content, which results in the phosphorylation of HSL Ser660. This adrenergic stimulation contributes to the increase in HSL activity that occurs in human skeletal muscle in the first minute of exercise at 65% and 90% VO2 peak.

  16. Triglyceride lipases alter fuel metabolism and mitochondrial gene expression.

    Science.gov (United States)

    Watt, Matthew J

    2009-06-01

    Fatty acids derived from the hydrolysis of adipose tissue and skeletal muscle triacylglycerol (TG) are an important energy substrate at rest and during prolonged moderate-intensity exercise. Hormone sensitive lipase (HSL) was long considered to be the rate-limiting enzyme for adipocyte and skeletal muscle TG lipolysis. However, the understanding of TG lipolysis regulation was recently challenged by the finding that adipose TG lipase (ATGL) is the predominant TG lipase in adipose tissue and an important regulator of TG degradation in skeletal muscle. Thus, it is now proposed that ATGL and HSL regulate lipolysis in a serial manner, with ATGL cleaving the first fatty acid and HSL the second fatty acid of TG. Further to this biochemical evaluation, the generation and metabolic characterization of ATGL-/- and HSL-/- mice have revealed distinct phenotypes. ATGL-/- mice are obese, exhibit impaired thermogenesis, oxidize more carbohydrate, and die prematurely due to cardiac dysfunction. Studies in HSL-/- mice report defective beta-adrenergic stimulated lipolysis, protection against high-fat diet-induced obesity, and possible impairments in insulin secretion. This review outlines the current understanding of the cellular regulation of TG lipases, lipolytic regulation, and the functional implications of manipulating ATGL and HSL in vivo.

  17. Relationship between site-specific HSL phosphorylation and adipocyte lipolysis in obese women.

    Science.gov (United States)

    Lorente-Cebrián, Silvia; Kulyté, Agné; Hedén, Per; Näslund, Erik; Arner, Peter; Rydén, Mikael

    2011-01-01

    In fat cells of obese humans, basal lipolysis is increased but catecholamine-stimulated lipolysis is blunted. This is linked to decreased expression of hormone-sensitive lipase (HSL). Upon stimulation by cAMP, HSL is phosphorylated at several serine residues (P-Ser(552), P-Ser(649) and P-Ser(650)) leading to enzymatic activation. In contrast, P-Ser(554) prevents phosphorylation at Ser(552) and is thus considered an inactivating site. We hypothesized that differences in HSL phosphorylation could be linked to disturbed adipocyte lipolysis in obesity. Phosphorylation at Ser(552), Ser(554), Ser(650) as well as total HSL and adipose triglyceride lipase (ATGL) protein expression were assessed by Western blot in subcutaneous adipose tissue samples of 32 obese women. Basal and stimulated lipolysis in isolated fat cells were correlated to phosphorylation levels. While there was no correlation between basal lipolysis and P-Ser(650) or P-Ser(554), there was a negative correlation with P-Ser(552) (r = 0.39; p lipolysis. There were no significant correlations between any measure of lipolysis and total levels of HSL and ATGL. In contrast to total HSL and ATGL levels, phosphorylation at Ser(554) and Ser(552), but not at Ser(650), may differentially predict adipocyte lipolysis in vitro. Posttranslational modifications of HSL may therefore constitute an important regulator of adipocyte lipolysis, at least in adipose tissue of obese women. Whether this is also relevant in lean individuals remains to be demonstrated. Copyright © 2011 S. Karger AG, Basel.

  18. Placental lipases in pregnancies complicated by gestational diabetes mellitus (GDM.

    Directory of Open Access Journals (Sweden)

    Helen L Barrett

    Full Text Available Infants of women with gestational diabetes mellitus (GDM are more likely to be born large for gestational age with a higher percentage body fat. Elevated maternal lipids may contribute to this. Placental lipases such as lipoprotein lipase (LPL, endothelial lipase (EL and hormone sensitive lipase (HSL are involved in transferring lipids from mother to fetus. Previous studies of expression of these lipases in placentae in women with diabetes in pregnancy have reported divergent results. Intracellular lipases such as adipose triglyceride lipase (ATGL, and HSL are central to lipid droplet metabolism. The activities of these lipases are both influenced by Perilipin 1, and ATGL is also activated by a co-factor comparative gene identification-58 (CGI-58 and inhibited by G0/G1 switch gene 2 (GS02. None of these modifying factors or ATGL have been examined previously in placenta. The purpose of this study was therefore to examine the expression of ATGL, HSL, LPL, EL, as well as Perilipin 1, GS02 and CGI-58 in term pregnancies complicated by GDM. mRNA and protein expression of the lipases were measured in placentae from 17 women with GDM and 17 normoglycaemic pregnancies, matched for maternal BMI and gestational age of delivery. ATGL mRNA expression was increased and HSL mRNA expression reduced in placentae from GDM although there was no differences in protein expression of any of the lipases. All lipases were localised to trophoblasts and endothelial cells. The expression of Perilipin 1 and CGI-58 mRNA was increased and GS02 not altered in GDM. These results suggest that there is no difference in expression in these four lipases between GDM and normoglycaemic placentae, and therefore altered lipid transfer via these lipases does not contribute to large for gestational age in infants of women with GDM.

  19. The regulation of HSL and LPL expression by DHT and flutamide in human subcutaneous adipose tissue.

    Science.gov (United States)

    Anderson, L A; McTernan, P G; Harte, A L; Barnett, A H; Kumar, S

    2002-05-01

    Clinical observations suggest a role for testosterone in the accumulation of central adiposity and with an associated increased risk of disease. To date, no human study has analysed the role of dihydrotestosterone (DHT) on adipose tissue mass regulation in vitro. This study investigated the role of DHT and androgen receptors (AR) in the regulation of lipolysis and lipogenesis by examining the key enzymes hormone sensitive lipase (HSL) and lipoprotein lipase (LPL) respectively. Isolated abdominal subcutaneous adipocytes (Scad) (n = 15) were treated with either DHT (10(-7)-10(-9) m), an antiandrogen, flutamide (FLT: 10(-7)-10(-9) m) or a combination of DHT (10(-7)-10(-9) m) with FLT (10(-8) m). Relative protein expression of HSL, LPL and AR was determined. In Scad, DHT inhibited HSL expression maximally at 10(-9) m (0.7 +/- 0.4**; p DHT10(-9) m (2.22 +/- 0.48*; p DHT + FLT compared with DHT alone. Androgen receptor expression studies showed an inverse correlation with DHT, whereas DHT + FLT reduced AR expression. These studies indicate that DHT may alter HSL and LPL expression, whereas only LPL expression appears mediated by AR. These findings suggest a physiological role for DHT in the control of adipose tissue mass in women, and indicate that androgens may also play an important role in regulating lipid metabolism.

  20. Piromelatine decreases triglyceride accumulation in insulin resistant 3T3-L1 adipocytes: role of ATGL and HSL.

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    Wang, Ping-Ping; She, Mei-Hua; He, Ping-Ping; Chen, Wu-Jun; Laudon, Moshe; Xu, Xuan-Xuan; Yin, Wei-Dong

    2013-08-01

    Piromelatine, a novel investigational multimodal sleep medicine, is developed for the treatment of patients with primary and co-morbid insomnia. Piromelatine has been shown to inhibit weight gain and improve insulin sensitivity in high-fat/high-sucrose-fed (HFHS) rats. Considering that piromelatine has also been implicated in lowering of triglyceride levels in HFHS rats, this work elucidated whether this effect involves in the regulation of adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (HSL) in triglyceride (TG) metabolism. In this study, we investigated the effects of piromelatine and MT2 receptors inhibition on TG content, insulin-stimulated glucose uptake, and the expressions of ATGL and HSL in 3T3-L1 adipocytes preincubated in high glucose and high insulin (HGI) conditions. Our results showed that culturing 3T3-L1 adipocytes under HGI conditions increased triglyceride accumulation with concomitant decrease of ATGL and HSL expression, inducing insulin resistance in 3T3-L1 adipocytes. We also found that triglyceride accumulation was significantly inhibited and the levels of ATGL/HSL increased after melatonin or piromelatine treatment. The effects of melatonin/piromelatine (10 nM) were counteracted by pretreatment with the relatively selective MT2 receptor antagonist luzindole (100 nM). In this study, our data demonstrate that piromelatine reverses high glucose and high insulin-induced triglyceride accumulation in 3T3-L1 adipocytes, possibly through up-regulating of ATGL and HSL expression via a melatonin-dependent manner.

  1. Resistance to high-fat diet-induced obesity and altered expression of adipose-specific genes in HSL-deficient mice.

    Science.gov (United States)

    Harada, Kenji; Shen, Wen-Jun; Patel, Shailja; Natu, Vanita; Wang, Jining; Osuga, Jun-ichi; Ishibashi, Shun; Kraemer, Fredric B

    2003-12-01

    To elucidate the role of hormone-sensitive lipase (HSL) in diet-induced obesity, HSL-deficient (HSL-/-) and wild-type mice were fed normal chow or high-fat diets. HSL-/- mice were resistant to diet-induced obesity showing higher core body temperatures. Weight and triacylglycerol contents were decreased in white adipose tissue (WAT) but increased in both brown adipose tissue (BAT) and liver of HSL-/- mice. Serum insulin levels in the fed state and tumor necrosis factor-alpha mRNA levels in adipose tissues were higher, whereas serum levels of adipocyte complement-related protein of 30 kDa (ACRP30)/adiponectin and leptin, as well as mRNA levels of ACRP30/adiponectin, leptin, resistin, and adipsin in WAT, were lower in HSL-/- mice than in controls. Expression of transcription factors associated with adipogenesis (peroxisome proliferator-activated receptor-gamma, CAAT/enhancer-binding protein-alpha) and lipogenesis (carbohydrate response element-binding protein, adipocyte determination- and differentiation-dependent factor-1/sterol regulatory element-binding protein-1c), as well as of adipose differentiation markers (adipocyte lipid-binding protein, perilipin, lipoprotein lipase), lipogenic enzymes (glycerol-3-phosphate acyltransferase, acyl-CoA:diacylglycerol acyltransferase-1 and -2, fatty acid synthase, ATP citrate lyase) and insulin signaling proteins (insulin receptor, insulin receptor substrate-1, GLUT4), was suppressed in WAT but not in BAT of HSL-/- mice. In contrast, expression of genes associated with cholesterol metabolism (sterol-regulatory element-binding protein-2, 3-hydroxy-3-methylglutaryl-CoA reductase, acyl-CoA:cholesterol acyltransferase-1) and thermogenesis (uncoupling protein-2) was upregulated in both WAT and BAT of HSL-/- mice. Our results suggest that impaired lipolysis in HSL deficiency affects lipid metabolism through alterations of adipose differentiation and adipose-derived hormone levels.

  2. Theophylline-Based KMUP-1 Improves Steatohepatitis via MMP-9/IL-10 and Lipolysis via HSL/p-HSL in Obese Mice.

    Science.gov (United States)

    Wu, Bin-Nan; Kuo, Kung-Kai; Chen, Yu-Hsun; Chang, Chain-Ting; Huang, Hung-Tu; Chai, Chee-Yin; Dai, Zen-Kong; Chen, Ing-Jun

    2016-08-17

    KMUP-1 (7-[2-[4-(2-chlorobenzene)piperazinyl]ethyl]-1,3-dimethylxanthine) has been reported to cause hepatic fat loss. However, the action mechanisms of KMUP-1 in obesity-induced steatohepatitis remains unclear. This study elucidated the steatohepatitis via matrix metallopeptidase 9 (MMP-9) and tumor necrosis factor α (TNFα), and related lipolysis via hormone sensitive lipase (HSL) and adipose triglyceride lipase (ATGL) by KMUP-1. KMUP-1 on steatohepatitis-associated HSL/p-HSL/ATGL/MMP-9/TNFα/interleukin-10 (IL-10) and infiltration of M1/M2 macrophages in obese mice were examined. KMUP-1 was administered by oral gavage from weeks 1-14 in high-fat diet (HFD)-supplemented C57BL/6J male mice (protection group) and from weeks 8-14, for 6 weeks, in HFD-induced obese mice (treatment group). Immunohistochemistry (IHC) and hematoxylin and eosin (H&E) staining of tissues, oil globules number and size, infiltration and switching of M1/M2 macrophages were measured to determine the effects on livers. IL-10 and MMP-9 proteins were explored to determine the effects of KMUP-1 on M1/M2 macrophage polarization in HFD-induced steatohepatitis. Long-term administration of KMUP-1 reversed HFD-fed mice increased in body weight, sGOT/sGPT, triglyceride (TG) and glucose. Additionally, KMUP-1 decreased MMP-9 and reactive oxygen species (ROS), and increased HSL/p-HSL and IL-10 in HFD mice livers. In conclusion, KMUP-1, a phosphodiesterase inhibitor (PDEI), was shown to reduce lipid accumulation in liver tissues, suggesting that it could be able to prevent or treat steatohepatitis induced by HFD.

  3. Theophylline-Based KMUP-1 Improves Steatohepatitis via MMP-9/IL-10 and Lipolysis via HSL/p-HSL in Obese Mice

    Directory of Open Access Journals (Sweden)

    Bin-Nan Wu

    2016-08-01

    Full Text Available KMUP-1 (7-[2-[4-(2-chlorobenzenepiperazinyl]ethyl]-1,3-dimethylxanthine has been reported to cause hepatic fat loss. However, the action mechanisms of KMUP-1 in obesity-induced steatohepatitis remains unclear. This study elucidated the steatohepatitis via matrix metallopeptidase 9 (MMP-9 and tumor necrosis factor α (TNFα, and related lipolysis via hormone sensitive lipase (HSL and adipose triglyceride lipase (ATGL by KMUP-1. KMUP-1 on steatohepatitis-associated HSL/p-HSL/ATGL/MMP-9/TNFα/interleukin-10 (IL-10 and infiltration of M1/M2 macrophages in obese mice were examined. KMUP-1 was administered by oral gavage from weeks 1–14 in high-fat diet (HFD-supplemented C57BL/6J male mice (protection group and from weeks 8–14, for 6 weeks, in HFD-induced obese mice (treatment group. Immunohistochemistry (IHC and hematoxylin and eosin (H&E staining of tissues, oil globules number and size, infiltration and switching of M1/M2 macrophages were measured to determine the effects on livers. IL-10 and MMP-9 proteins were explored to determine the effects of KMUP-1 on M1/M2 macrophage polarization in HFD-induced steatohepatitis. Long-term administration of KMUP-1 reversed HFD-fed mice increased in body weight, sGOT/sGPT, triglyceride (TG and glucose. Additionally, KMUP-1 decreased MMP-9 and reactive oxygen species (ROS, and increased HSL/p-HSL and IL-10 in HFD mice livers. In conclusion, KMUP-1, a phosphodiesterase inhibitor (PDEI, was shown to reduce lipid accumulation in liver tissues, suggesting that it could be able to prevent or treat steatohepatitis induced by HFD.

  4. Theophylline-Based KMUP-1 Improves Steatohepatitis via MMP-9/IL-10 and Lipolysis via HSL/p-HSL in Obese Mice

    Science.gov (United States)

    Wu, Bin-Nan; Kuo, Kung-Kai; Chen, Yu-Hsun; Chang, Chain-Ting; Huang, Hung-Tu; Chai, Chee-Yin; Dai, Zen-Kong; Chen, Ing-Jun

    2016-01-01

    KMUP-1 (7-[2-[4-(2-chlorobenzene)piperazinyl]ethyl]-1,3-dimethylxanthine) has been reported to cause hepatic fat loss. However, the action mechanisms of KMUP-1 in obesity-induced steatohepatitis remains unclear. This study elucidated the steatohepatitis via matrix metallopeptidase 9 (MMP-9) and tumor necrosis factor α (TNFα), and related lipolysis via hormone sensitive lipase (HSL) and adipose triglyceride lipase (ATGL) by KMUP-1. KMUP-1 on steatohepatitis-associated HSL/p-HSL/ATGL/MMP-9/TNFα/interleukin-10 (IL-10) and infiltration of M1/M2 macrophages in obese mice were examined. KMUP-1 was administered by oral gavage from weeks 1–14 in high-fat diet (HFD)-supplemented C57BL/6J male mice (protection group) and from weeks 8–14, for 6 weeks, in HFD-induced obese mice (treatment group). Immunohistochemistry (IHC) and hematoxylin and eosin (H&E) staining of tissues, oil globules number and size, infiltration and switching of M1/M2 macrophages were measured to determine the effects on livers. IL-10 and MMP-9 proteins were explored to determine the effects of KMUP-1 on M1/M2 macrophage polarization in HFD-induced steatohepatitis. Long-term administration of KMUP-1 reversed HFD-fed mice increased in body weight, sGOT/sGPT, triglyceride (TG) and glucose. Additionally, KMUP-1 decreased MMP-9 and reactive oxygen species (ROS), and increased HSL/p-HSL and IL-10 in HFD mice livers. In conclusion, KMUP-1, a phosphodiesterase inhibitor (PDEI), was shown to reduce lipid accumulation in liver tissues, suggesting that it could be able to prevent or treat steatohepatitis induced by HFD. PMID:27548140

  5. [Adipose triglyceride lipase regulates adipocyte lipolysis].

    Science.gov (United States)

    Xu, Chong; Xu, Guo-Heng

    2008-01-01

    Obesity, insulin resistance, and type 2 diabetes are associated with elevated concentration of circulating free fatty acids (FFAs), which are critically governed by the process of triglyceride lipolysis in adipocytes. Hormone-sensitive lipase (HSL) and adipose triglyceride lipase (ATGL) are two major enzymes in the control of triacylglycerol hydrolysis in adipose tissue. ATGL expressed predominantly in white adipose tissue specifically initiates triacylglycerol hydrolysis to generate diacylglycerols and FFA, a role distinguished from HSL that mainly hydrolyzes diacylglycerols. The transcription of ATGL is regulated by several factors. ATGL activity is regulated by CGI-58. Under basal conditions, interaction of CGI-58 with a lipid droplet associating protein, perilipin, results in an inactivation of ATGL activity. During PKA-stimulated lipolysis, CGI-58 is released from phosphorylated perilipin and in turn, binds to ATGL. This action facilitates triglyceride lipolysis. This review focuses on the regulation and function of ATGL in adipose lipolysis and metabolism.

  6. Adipose triglyceride lipase contributes to cancer-associated cachexia.

    Science.gov (United States)

    Das, Suman K; Eder, Sandra; Schauer, Silvia; Diwoky, Clemens; Temmel, Hannes; Guertl, Barbara; Gorkiewicz, Gregor; Tamilarasan, Kuppusamy P; Kumari, Pooja; Trauner, Michael; Zimmermann, Robert; Vesely, Paul; Haemmerle, Guenter; Zechner, Rudolf; Hoefler, Gerald

    2011-07-08

    Cachexia is a multifactorial wasting syndrome most common in patients with cancer that is characterized by the uncontrolled loss of adipose and muscle mass. We show that the inhibition of lipolysis through genetic ablation of adipose triglyceride lipase (Atgl) or hormone-sensitive lipase (Hsl) ameliorates certain features of cancer-associated cachexia (CAC). In wild-type C57BL/6 mice, the injection of Lewis lung carcinoma or B16 melanoma cells causes tumor growth, loss of white adipose tissue (WAT), and a marked reduction of gastrocnemius muscle. In contrast, Atgl-deficient mice with tumors resisted increased WAT lipolysis, myocyte apoptosis, and proteasomal muscle degradation and maintained normal adipose and gastrocnemius muscle mass. Hsl-deficient mice with tumors were also protected although to a lesser degree. Thus, functional lipolysis is essential in the pathogenesis of CAC. Pharmacological inhibition of metabolic lipases may help prevent cachexia.

  7. Fat-reducing effects of dehydroepiandrosterone involve upregulation of ATGL and HSL expression, and stimulation of lipolysis in adipose tissue.

    Science.gov (United States)

    Karbowska, Joanna; Kochan, Zdzislaw

    2012-11-01

    Dehydroepiandrosterone (DHEA) reduces body fat in rodents and humans, and increases glycerol release from isolated rat epididymal adipocytes and human visceral adipose tissue explants. It suggests that DHEA stimulates triglyceride hydrolysis in adipose tissue; however, the mechanisms underlying this action are still unclear. We examined the effects of DHEA on the expression of adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (HSL), the key enzymes of lipolysis, in rat epididymal white adipose tissue (eWAT). Male Wistar rats were fed a diet containing 0.6% DHEA for 2 weeks and eWAT was analyzed for mRNA and protein expression of ATGL and HSL, as well as mRNA expression of peroxisome proliferator-activated receptor γ 2 (PPARγ2) and its downstream target fatty acid translocase (FAT). Glycerol release from eWAT explants and serum free fatty acids (FFA) were also measured. Rats that received DHEA gained less weight, had 23% lower eWAT mass and 31% higher serum FFA levels than controls. Cultured explants of eWAT from DHEA-treated rats released 81% more glycerol than those from control rats. DHEA administration upregulated ATGL mRNA (1.62-fold, Padipose tissue by increasing the expression and activity of ATGL and HSL. The effects of DHEA appear to be mediated, at least in part, via PPARγ2 activation, which in turn upregulates ATGL and HSL gene expression. Copyright © 2012 Elsevier Inc. All rights reserved.

  8. Decrease in intramuscular lipid droplets and translocation of HSL in response to muscle contraction and epinephrine.

    Science.gov (United States)

    Prats, Clara; Donsmark, Morten; Qvortrup, Klaus; Londos, Constantine; Sztalryd, Carole; Holm, Cecilia; Galbo, Henrik; Ploug, Thorkil

    2006-11-01

    A better understanding of skeletal muscle lipid metabolism is needed to identify the molecular mechanisms relating intramuscular triglyceride (IMTG) to muscle metabolism and insulin sensitivity. An increasing number of proteins have been reported to be associated with intracellular triglyceride (TG), among them the PAT family members: perilipin, ADRP (for adipocyte differentiation-related protein), and TIP47 (for tail-interacting protein of 47 kDa). Hormone-sensitive lipase (HSL) is thought to be the major enzyme responsible for IMTG hydrolysis in skeletal muscle. In adipocytes, regulation of HSL by intracellular redistribution has been demonstrated. The existence of such regulatory mechanisms in skeletal muscle has long been hypothesized but has never been demonstrated. The aim of this study was to characterize the PAT family proteins associated with IMTG and to investigate the effect of epinephrine stimulation or muscle contraction on skeletal muscle TG content and HSL intracellular distribution. Rat soleus muscles were either incubated with epinephrine or electrically stimulated for 15 min. Single muscle fibers were used for morphological analysis by confocal and transmission electron microscopy. We show a decrease in IMTG in response to both lipolytic stimuli. Furthermore, we identify two PAT family proteins, ADRP and TIP47, associated with IMTG. Finally, we demonstrate HSL translocation to IMTG and ADRP after stimulation with epinephrine or contraction.

  9. Chronic TNFalpha and cAMP pre-treatment of human adipocytes alter HSL, ATGL and perilipin to regulate basal and stimulated lipolysis.

    Science.gov (United States)

    Bézaire, Véronic; Mairal, Aline; Anesia, Rodica; Lefort, Corinne; Langin, Dominique

    2009-09-17

    We examined the effects of chronic TNFalpha and dibutyryl-cAMP (Db-cAMP) pre-treatment on the lipolytic machinery of human hMADS adipocytes. TNFalpha decreased adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (HSL) protein content and triglycerides (TG)-hydrolase activity but increased basal lipolysis due to a marked reduction in perilipin (PLIN) protein content. Conversely, Db-cAMP increased ATGL and HSL protein content but prevented PLIN phosphorylation, the net result being accentuated basal lipolysis. In forskolin-stimulated conditions, TNFalpha and Db-cAMP pre-treatment decreased stimulated TG-hydrolase activity and impaired PLIN phosphorylation. Together, this resulted in a severely attenuated response to forskolin-stimulated lipolysis.

  10. Beta-adrenergic stimulation of skeletal muscle HSL can be overridden by AMPK signaling.

    Science.gov (United States)

    Watt, Matthew J; Steinberg, Gregory R; Chan, Stanley; Garnham, Andrew; Kemp, Bruce E; Febbraio, Mark A

    2004-09-01

    Hormone-sensitive lipase (HSL), an important regulatory enzyme for triacylglycerol hydrolysis within skeletal muscle, is controlled by beta-adrenergic signaling as well as intrinsic factors related to contraction and energy turnover. In the current study, we tested the capacity of 5'AMP-activated protein kinase (AMPK) to suppress beta-adrenergic stimulation of HSL activity. Eight male subjects completed 60 min of cycle exercise at 70% VO2 peak on two occasions: either with normal (CON) or low (LG) pre-exercise muscle glycogen content, which is known to enhance exercise-induced AMPK activity. Muscle samples were obtained before and immediately after exercise. Pre-exercise glycogen averaged 375 +/- 35 and 163 +/- 27 mmol x kg(-1) dm for CON and LG, respectively. AMPK alpha-2 was not different between trials at rest and was increased (3.7-fold, PHSL activity did not differ between trials at rest and increased (0 min: 1.67 +/- 0.13; 60 min: 2.60 +/- 0.26 mmol x min(-1) x kg(-1) dm) in CON. The exercise-induced increase in HSL activity was attenuated by AMPK alpha-2 activation in LG. The attenuated HSL activity during LG occurred despite higher plasma epinephrine levels (60 min: CON, 1.96 +/- 0.29 vs LG, 4.25 +/- 0.60 nM, PHSL activity in LG, IMTG was decreased by exercise (0 min: 27.1 +/- 2.0; 60 min: 22.5 +/- 2.0 mmol x kg(-1) dm, PHSL activity, we performed experiments in muscle cell culture. The epineprine-induced increase in HSL activity was totally attenuated (PHSL activity that can override beta-adrenergic stimulation. However, the increased IMTG degradation in LG suggests factors other than HSL activity are important for IMTG degradation.

  11. Developmental changes of the FAS and HSL mRNA expression and their effects on the content of intramuscular fat in Kazak and Xinjiang sheep.

    Science.gov (United States)

    Qiao, Yong; Huang, Zhiguo; Li, Qifa; Liu, Zhenshan; Hao, Chengli; Shi, Guoqing; Dai, Rong; Xie, Zhuang

    2007-10-01

    Twenty-four male Kazak sheep and 30 Xinjiang fine wool sheep at different ages were selected to investigate the development-dependent expression levels of fatty acid synthase (FAS) gene and hormone-sensitive lipase (HSL) gene in muscle and their effects on the contents of intramuscular fat (IMF). Longissimus dorsal muscle was sampled to measure IMF and total RNA was extracted to determine FAS and HSL mRNA expression levels by real-time PCR. The results showed that: 1) The IMF content increased continuously with growth and showed significant differences (P HSL mRNA expression level had a similar model in two breeds, in Kazak sheep it was the highest on day 0 (P HSL mRNA expression level were both negatively related to IMF content (r = -0.485 (P = 0.02), r = -0.423 (P = 0.05)), and the ratio of FAS/HSL expression exhibited significantly negatively related IMF contents. In male Xinjiang sheep, there were no obvious relationship between FAS and HSL expression and IMF content (P > 0.05).

  12. Resveratrol regulates lipolysis via adipose triglyceride lipase.

    Science.gov (United States)

    Lasa, Arrate; Schweiger, Martina; Kotzbeck, Petra; Churruca, Itziar; Simón, Edurne; Zechner, Rudolf; Portillo, María del Puy

    2012-04-01

    Resveratrol has been reported to increase adrenaline-induced lipolysis in 3T3-L1 adipocytes. The general aim of the present work was to gain more insight concerning the effects of trans-resveratrol on lipid mobilization. The specific purpose was to assess the involvement of the two main lipases: adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (HSL), in the activation of lipolysis induced by this molecule. For lipolysis experiments, 3T3-L1 and human SGBS adipocytes as well as adipose tissue from wild-type, ATGL knockout and HSL knockout mice were used. Moreover, gene and protein expressions of these lipases were analyzed. Resveratrol-induced free fatty acids release but not glycerol release in 3T3-L1 under basal and isoproterenol-stimulating conditions and under isoproterenol-stimulating conditions in SGBS adipocytes. When HSL was blocked by compound 76-0079, free fatty acid release was still induced by resveratrol. By contrast, in the presence of the compound C, an inhibitor of adenosine monophosphate-activated protein kinase, resveratrol effect was totally blunted. Resveratrol increased ATGL gene and protein expressions, an effect that was not observed for HSL. Resveratrol increased fatty acids release in epididymal adipose tissue from wild-type and HSL knockout mice but not in that adipose tissue from ATGL knockout mice. Taking as a whole, the present results provide novel evidence that resveratrol regulates lipolytic activity in human and murine adipocytes, as well as in white adipose tissue from mice, acting mainly on ATGL at transcriptional and posttranscriptional levels. Enzyme activation seems to be induced via adenosine monophosphate-activated protein kinase. Copyright © 2012 Elsevier Inc. All rights reserved.

  13. Lipolysis and lipases in white adipose tissue - An update.

    Science.gov (United States)

    Bolsoni-Lopes, Andressa; Alonso-Vale, Maria Isabel C

    2015-08-01

    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.

  14. Dysregulation of lipolysis and lipid metabolism in visceral and subcutaneous adipocytes by high-fat diet: role of ATGL, HSL, and AMPK.

    Science.gov (United States)

    Gaidhu, Mandeep P; Anthony, Nicole M; Patel, Prital; Hawke, Thomas J; Ceddia, Rolando B

    2010-04-01

    This study investigated the molecular mechanisms by which a high-fat diet (HFD) dysregulates lipolysis and lipid metabolism in mouse epididymal (visceral, VC) and inguinal (subcutaneous, SC) adipocytes. Eight-weeks of HFD feeding increased adipose triglyceride lipase (ATGL) content and comparative gene identification-58 (CGI-58) expression, whereas hormone-sensitive lipase (HSL) phosphorylation and perilipin content were severely reduced. Adipocytes from HFD mice elicited increased basal but blunted epinephrine-stimulated lipolysis and increased diacylglycerol content in both fat depots. Consistent with impaired adrenergic receptor signaling, HFD also increased adipose-specific phospholipase A(2) expression in both fat depots. Inhibition of E-prostanoid 3 receptor increased basal lipolysis in control adipocytes but failed to acutely alter the effects of HFD on lipolysis in both fat depots. In HFD visceral adipocytes, activation of adenylyl cyclases by forskolin increased HSL phosphorylation and surpassed the lipolytic response of control cells. However, in HFD subcutaneous adipocytes, forskolin induced lipolysis without detectable HSL phosphorylation, suggesting activation of an alternative lipase in response to HFD-induced suppression of HSL in VC and SC adipocytes. HFD also powerfully inhibited basal, epinephrine-, and forskolin-induced AMP kinase (AMPK) activation as well peroxisome proliferator-activated receptor gamma coactivator-1alpha expression, citrate synthase activity, and palmitate oxidation in both fat depots. In summary, novel evidence is provided that defective adrenergic receptor signaling combined with upregulation of ATGL and suppression of HSL and AMPK signaling mediate HFD-induced alterations in lipolysis and lipid utilization in VC and SC adipocytes, which may play an important role in defective lipid mobilization and metabolism seen in diet-induced obesity.

  15. Association of HSL gene E1-c.276C>T and E8-c.51C>T mutation with economical traits of Chinese Simmental cattle.

    Science.gov (United States)

    Fang, X B; Zhang, L P; Yu, X Z; Li, J Y; Lu, C Y; Zhao, Z H; Yang, R J

    2014-01-01

    Hormone-sensitive lipase (HSL) is responsible for the decomposition of triglycerides in adipose tissue to release free fatty acids, and it is a key rate-limiting enzyme in the regulation of adipose tissue deposition and decomposition. The objective of this study was to evaluate the association between novel SNPs in the coding region of bovine HSL gene and carcass and meat quality traits of Chinese Simmental-cross steers. Two novel SNPs were genotyped and the 47 traits of carcass and meat quality traits were measured in the population studied. Statistical analysis revealed that the SNPs of HSL gene were associated with the carcass and meat quality traits. The individuals with TT genotypes of E1-276C>T showed significant higher dressing percentage, net meat rate, hind legs circumference, fat coverage rate, mesenteric fat and kidney fat (p T (P17S) also showed a significant association with the pH of beef and fatty acids content in Chinese Simmental cattle (p HSL might be one of important genetic factors that influence carcass yield and meat quality in beef cattle, and it may be a useful marker for meat quality traits in future marker-assisted selection programs in beef cattle breeding and production.

  16. Prolonged AICAR-induced AMP-kinase activation promotes energy dissipation in white adipocytes: novel mechanisms integrating HSL and ATGL.

    Science.gov (United States)

    Gaidhu, Mandeep P; Fediuc, Sergiu; Anthony, Nicole M; So, Mandy; Mirpourian, Mani; Perry, Robert L S; Ceddia, Rolando B

    2009-04-01

    This study was designed to investigate the effects of prolonged activation of AMP-activated protein kinase (AMPK) on lipid partitioning and the potential molecular mechanisms involved in these processes in white adipose tissue (WAT). Rat epididymal adipocytes were incubated with 5'-aminoimidasole-4-carboxamide-1-beta-d-ribofuranoside (AICAR;0.5 mM) for 15 h. Also, epididymal adipocytes were isolated 15 h after AICAR was injected (i.p. 0.7 g/kg body weight) in rats. Adipocytes were utilized for various metabolic assays and for determination of gene expression and protein content. Time-dependent in vivo plasma NEFA concentrations were determined. AICAR treatment significantly increased AMPK activation, inhibited lipogenesis, and increased FA oxidation. This was accompanied by upregulation of peroxisome proliferator-activated receptor (PPAR)alpha, PPARdelta, and PPARgamma-coactivator-1alpha (PGC-1alpha) mRNA levels. Lipolysis was first suppressed, but then increased, both in vitro and in vivo, with prolonged AICAR treatment. Exposure to AICAR increased adipose triglyceride lipase (ATGL) content and FA release, despite inhibition of basal and epinephrine-stimulated hormone-sensitive lipase (HSL) activity. Here, we provide evidence that prolonged AICAR-induced AMPK activation can remodel adipocyte metabolism by upregulating pathways that favor energy dissipation versus lipid storage in WAT. Additionally, we show novel time-dependent effects of AICAR-induced AMPK activation on lipolysis, which involves antagonistic modulation of HSL and ATGL.

  17. Changes in the expression of HSL and OCTN2 in the female reproductive tract of the bat, Scotophilus heathii in relation to sperm storage.

    Science.gov (United States)

    Roy, Vikas Kumar; Krishna, Amitabh

    2012-07-01

    The aim of this study was to compare the changes in the expression of hormone sensitive lipase (HSL) and organic cation transporter 2 (OCTN2) in different regions of the female reproductive tract of bats (Scotophilus heathii) during the period of sperm storage. Western blot analysis revealed that both HSL and OCTN2 showed elevated expression in the utero-tubal junction, the site of sperm storage, compared to the vagina, cervix and uterus. An immunohistochemical study showed localization of HSL and OCTN2 in the cytoplasm of luminal epithelial cells in the utero-tubal junction of S. heathii, which suggests a role of fatty acids metabolism at the site of sperm storage. Furthermore, this study showed the presence OCTN2 immunostaining in the tail of stored sperm, which suggests a direct role of carnitine in sperm physiology. However, the role of carnitine in sperm storage needs further investigation. It is hypothesized that the utero-tubal junction may be finely tuned with fat depletion to support sperm storage in the female genital tract of S. heathii.

  18. Cardiotrophin-1 stimulates lipolysis through the regulation of main adipose tissue lipases[S

    Science.gov (United States)

    López-Yoldi, Miguel; Fernández-Galilea, Marta; Laiglesia, Laura M.; Larequi, Eduardo; Prieto, Jesús; Martínez, J. Alfredo; Bustos, Matilde; Moreno-Aliaga, Maria J.

    2014-01-01

    Cardiotrophin-1 (CT-1) is a cytokine with antiobesity properties and with a role in lipid metabolism regulation and adipose tissue function. The aim of this study was to analyze the molecular mechanisms involved in the lipolytic actions of CT-1 in adipocytes. Recombinant CT-1 (rCT-1) effects on the main proteins and signaling pathways involved in the regulation of lipolysis were evaluated in 3T3-L1 adipocytes and in mice. rCT-1 treatment stimulated basal glycerol release in a concentration- and time-dependent manner in 3T3-L1 adipocytes. rCT-1 (20 ng/ml for 24 h) raised cAMP levels, and in parallel increased protein kinase (PK)A-mediated phosphorylation of perilipin and hormone sensitive lipase (HSL) at Ser660. siRNA knock-down of HSL or PKA, as well as pretreatment with the PKA inhibitor H89, blunted the CT-1-induced lipolysis, suggesting that the lipolytic action of CT-1 in adipocytes is mainly mediated by activation of HSL through the PKA pathway. In ob/ob mice, acute rCT-1 treatment also promoted PKA-mediated phosphorylation of perilipin and HSL at Ser660 and Ser563, and increased adipose triglyceride lipase (desnutrin) content in adipose tissue. These results showed that the ability of CT-1 to regulate the activity of the main lipases underlies the lipolytic action of this cytokine in vitro and in vivo, and could contribute to CT-1 antiobesity effects. PMID:25351614

  19. 基于机器学习方法的激素敏感脂肪酶抑制剂活性预测%Activity Prediction of Hormone-Sensitive Lipase Inhibitors Based on Machine Learning Methods

    Institute of Scientific and Technical Information of China (English)

    吕巍; 薛英

    2010-01-01

    脂肪组织中,激素敏感脂肪酶(HSL)被认为是调节脂肪酸代谢的关键限速酶.HSL在糖尿病的发病过程中起重要作用,抑制HSL活性有助于糖尿病的治疗,因此探索新颖的HSL抑制剂成为当前研究的热门.在激素敏感脂肪酶的作用机制和三维结构缺乏的情况下,需要发展预测HSL抑制剂的方法.本文采用几种机器学习方法(支持向量机(SVM)、k-最近相邻法(k-NN)和C4.5决策树(C4.5 DT))对已知的HSL抑制剂与非抑制剂建市分类预测模型.252个结构多样性化合物(123个HSL抑制剂与129个HSL非抑制剂)被用于测试分类预测系统,并用递归变量消除法选择与HSL抑制剂相关的性质描述符以提高预测精度.本研究对独立验证集的总预测精度为75.0%-80.0%,HSL抑制剂的预测精度为85.7%-90.5%,非HSL抑制剂的预测精度为63.2%-68.4%.支持向量机方法给出最好的总预测精度(80.0%).本研究表明支持向量机等机器学习方法可以有效预测未知数据集中潜在的HSL抑制剂,并有助于发现与其相关的分子描述符.

  20. Leu452His mutation in lipoprotein lipase gene transfer associated with hypertriglyceridemia in mice in vivo.

    Directory of Open Access Journals (Sweden)

    Kaiyue Sun

    Full Text Available Mutated mouse lipoprotein lipase (LPL containing a leucine (L to histidine (H substitution at position 452 was transferred into mouse liver by hydrodynamics-based gene delivery (HD. Mutated-LPL (MLPL gene transfer significantly increased the concentrations of plasma MLPL and triglyceride (TG but significantly decreased the activity of plasma LPL. Moreover, the gene transfer caused adiposis hepatica and significantly increased TG content in mouse liver. To understand the effects of MLPL gene transfer on energy metabolism, we investigated the expression of key functional genes related to energy metabolism in the liver, epididymal fat, and leg muscles. The mRNA contents of hormone-sensitive lipase (HSL, adipose triglyceride lipase (ATGL, fatty acid-binding protein (FABP, and uncoupling protein (UCP were found to be significantly reduced. Furthermore, we investigated the mechanism by which MLPL gene transfer affected fat deposition in the liver, fat tissue, and muscle. The gene expression and protein levels of forkhead Box O3 (FOXO3, AMP-activated protein kinase (AMPK, and peroxisome proliferator-activated receptor-gamma coactivator 1 alpha (PGC-1α were found to be remarkably decreased in the liver, fat and muscle. These results suggest that the Leu452His mutation caused LPL dysfunction and gene transfer of MLPL in vivo produced resistance to the AMPK/PGC-1α signaling pathway in mice.

  1. PPARgamma agonism increases rat adipose tissue lipolysis, expression of glyceride lipases, and the response of lipolysis to hormonal control.

    Science.gov (United States)

    Festuccia, W T; Laplante, M; Berthiaume, M; Gélinas, Y; Deshaies, Y

    2006-10-01

    The aim of this study was to investigate the effect and mechanisms of action of in vivo peroxisome proliferator-activated receptor gamma (PPARgamma) activation on white adipose tissue (WAT) lipolysis and NEFA metabolism. Study rats were treated for 7 days with 15 mg/kg of rosiglitazone per day; control rats were not treated. After a 6-h fast, lipolysis and levels of mRNA for lipases were assessed in explants from various adipose depots. Rosiglitazone markedly increased basal and noradrenaline (norepinephrine)-stimulated glycerol and NEFA release from WAT explants, and amplified their inhibition by insulin. Primary adipocytes isolated from PPARgamma agonist-treated rats were also more responsive to noradrenaline stimulation expressed per cell, ruling out a contribution of an altered number of mature adipocytes in explants. Rosiglitazone concomitantly increased levels of mRNA transcripts for adipose triglyceride lipase (ATGL) and monoglyceride lipase (MGL) in subcutaneous and visceral WAT, and mRNA for hormone-sensitive lipase (HSL) in subcutaneous WAT. Lipase expression increased within 12 h of in vitro exposure of naïve explants to rosiglitazone, suggesting direct transcriptional activation. In parallel, chronic in vivo treatment with rosiglitazone lowered plasma NEFAs and in WAT its expected stimulatory action on glycerol and NEFA recycling, and on the expression of genes involved in NEFA uptake and retention by WAT, such processes counteracting net NEFA export. These findings demonstrate that, in the face of its plasma NEFA-lowering action, PPARgamma agonism stimulates WAT lipolysis, an effect that is compensated by lipid-retaining pathways. The results further suggest that PPARgamma agonism stimulates lipolysis by increasing the lipolytic potential, including the expression levels of the genes encoding adipose triglyceride lipase and monoglyceride lipase.

  2. Analysis of thermal adaptation in the HSL enzyme family.

    Science.gov (United States)

    Mandrich, L; Pezzullo, M; Del Vecchio, P; Barone, G; Rossi, M; Manco, G

    2004-01-01

    The recently solved three-dimensional (3D) structures of two thermostable members of the carboxylesterase/lipase HSL family, namely the Alicyclobacillus (formerly Bacillus) acidocaldarius and Archaeoglobus fulgidus carboxylesterases (EST2 and AFEST, respectively) were compared with that of the mesophilic homologous counterpart Brefeldine A esterase from Bacillus subtilis. Since the 3D homology models of other members of the HSL family were also available, we performed a structural alignment with all these sequences. The resulting alignment was used to assess the amino acid "traffic rule" in the HSL family. Quite surprisingly, the data were in very good agreement with those recently reported from two independent groups and based on the comparison of a huge number of homologous sequences from the genus Bacillus, Methanococcus and Deinococcus/Thermus. Taken as a whole, the data point to the statistical meaning of defined amino acid conversions going from psychrophilic to hyperthermophilic sequences. We identified and mapped several such changes onto the EST2 structure and observed that such mutations were localized mostly in loops regions or alpha-helices and were mostly excluded from the active site. A site-directed mutagenesis of two of the identified residues confirmed they were involved in thermal stability.

  3. Neu-p11 ameliorates insulin resisitance in 3T3-L1 adipocytes based on ATGL/HSL and its underlying mechanism%ATGL/HSL角度下解析Neu-p11改善胰岛素抵抗作用机制

    Institute of Scientific and Technical Information of China (English)

    王平平; 佘美华; Laudon Moshe; 尹卫东

    2013-01-01

    Aim To explore the possible role of adipose tissue triglyceride enzyme ( ATGL ) and hormonesensitive lipase ( HSL ) in high glucose and insulin ( HGI ) - induced insulin resistance in 3T3-L1 adipocytes and the underlying mechanisms. Methods 3T3-L1 adipocytes were administered with HGI for 24 h to induce insulin resistance. Glucose uptake and the quantitative determination of triglycerides were designed for detection indicators. Protein expressions were detected by Western blot. Results HGI incubating resulted in decreased insulin-stimulated glucose uptake and a significant increase in TG content in fat cells, with a concomitant decrease in ATGL and HSL protein expression. The Neu-p11 intervention reversed the effects of HGI on fat cells, while luzindole counteracted the effect of Neu-pll. Conclusions Neu-p11 might inhibit TG deposition in insulin-resistant 3T3-L1 adipocytes via MT2 receptor -dependent manner, at least in part by increasing triglyceride hydrolysis, resulting from enhancing ATGL and HSL levels.%目的 探讨脂肪组织甘油三酯酶(adipose triglyceride lipase,ATGL)及激素敏感性脂肪酶(hormone-sensitive lipase,HSL)在褪黑素非选择性受体激动剂Neu-p11改善高糖高胰岛素(high glucose and insulin,HGI)诱导的3T3-L1脂肪细胞胰岛素抵抗(insulin resistance,IR)中的作用及机制.方法 培养3T3-L1脂肪细胞,HGI诱导IR模型.以葡萄糖消耗量及细胞内甘油三酯(triglyceride,TG)定量测定作为检测指标,Western blot检测蛋白水平的表达情况.结果 HGI孵育减少脂肪细胞葡萄糖摄取,促进细胞内TG积聚,同时伴有ATGL及HSL的蛋白表达下调.Neu-p11干预逆转了HGI对脂肪细胞的作用效应,而MT2竞争性拮抗剂luzindole却拮抗了Neu-p11的上述效应.结论 Neu-p11以MT2受体依赖性方式抑制IR脂肪细胞TG沉积,可能与其上调ATGL、HSL蛋白的表达,促进TG水解相关.

  4. Leishmania donovani HslV does not interact stably with HslU proteins.

    Science.gov (United States)

    Chrobak, Mareike; Förster, Sabine; Meisel, Sarah; Pfefferkorn, Roxana; Förster, Frank; Clos, Joachim

    2012-04-01

    Genes for HslVU-type peptidases are found in bacteria and in a few select Eukaryota, among those such important pathogens as Plasmodium spp. and Leishmania spp. In this study, we performed replacements of all three HslV/HslU gene homologues and found one of those, HslV, to be essential for Leishmania donovani viability. The Leishmania HslV gene can also partially relieve the thermosensitive phenotype of a combined HslVU/Lon/ClpXP knockout mutant of Escherichia coli, indicating a conserved function. However, we found that the role and function of the two Leishmania HslU genes has diverged since neither of those interacts stably with HslV. The latter forms a dodecameric complex by itself and shows a punctate distribution. We conclude that whilst the basic function of HslV may be conserved in Leishmania, its organisation and interaction with its canonical complex partner HslU is not. Nevertheless, given the absence of HslV from the proteome of mammals and its essential role in Leishmania viability, HslV is a promising target for intervention.

  5. Xanthine-based KMUP-1 improves HDL via PPARγ/SR-B1, LDL via LDLRs, and HSL via PKA/PKG for hepatic fat loss.

    Science.gov (United States)

    Kuo, Kung-Kai; Wu, Bin-Nan; Liu, Chung-Pin; Yang, Tzu-Yang; Kao, Li-Pin; Wu, Jiunn-Ren; Lai, Wen-Ter; Chen, Ing-Jun

    2015-11-01

    The phosphodiesterase inhibitor (PDEI)/eNOS enhancer KMUP-1, targeting G-protein coupled receptors (GPCRs), improves dyslipidemia. We compared its lipid-lowering effects with simvastatin and explored hormone-sensitive lipase (HSL) translocation in hepatic fat loss. KMUP-1 HCl (1, 2.5, and 5 mg/kg/day) and simvastatin (5 mg/kg/day) were administered in C57BL/6J male mice fed a high-fat diet (HFD) by gavage for 8 weeks. KMUP-1 inhibited HFD-induced plasma/liver TG, total cholesterol, and LDL; increased HDL/3-hydroxy-3-methylglutaryl-CoA reductase (HMGR)/Rho kinase II (ROCK II)/PPARγ/ABCA1; and decreased liver and body weight. KMUP-1 HCl in drinking water (2.5 mg/200 ml tap water) for 1-14 or 8-14 weeks decreased HFD-induced liver and body weight and scavenger receptor class B type I expression and increased protein kinase A (PKA)/PKG/LDLRs/HSL expression and immunoreactivity. In HepG2 cells incubated with serum or exogenous mevalonate, KMUP-1 (10(-7)∼10(-5) M) reversed HMGR expression by feedback regulation, colocalized expression of ABCA1/apolipoprotein A-I/LXRα/PPARγ, and reduced exogenous geranylgeranyl pyrophosphate/farnesyl pyrophosphate (FPP)-induced RhoA/ROCK II expression. A guanosine 3',5'-cyclic monophosphate (cGMP) antagonist reversed KMUP-1-induced ROCK II reduction, indicating cGMP/eNOS involvement. KMUP-1 inceased PKG and LDLRs surrounded by LDL and restored oxidized LDL-induced PKA expresion. Unlike simvastatin, KMUP-1 could not inhibit (14)C mevalonate formation. KMUP-1 could, but simvastatin could not, decrease ROCK II expression by exogenous FPP/CGPP. KMUP-1 improves HDL via PPARγ/LXRα/ABCA1/Apo-I expression and increases LDLRs/PKA/PKG/HSL expression and immunoreactivity, leading to TG hydrolysis to lower hepatic fat and body weight.

  6. Adipose triglyceride lipase expression and fasting regulation are differently affected by cold exposure in adipose tissues of lean and obese Zucker rats.

    Science.gov (United States)

    Caimari, Antoni; Oliver, Paula; Palou, Andreu

    2012-09-01

    Adipose triglyceride lipase (ATGL) hydrolyzes triacylglycerols to diacylglycerols in the first step of lipolysis, providing substrates for hormone-sensitive lipase (HSL). Here we studied whether ATGL messenger RNA (mRNA) and protein levels were affected by 24-h cold exposure in different white adipose tissue depots and in interscapular brown adipose tissue of lean and obese Zucker rats submitted to feeding and 14-h fasting conditions. HSL mRNA expression was also studied in selected depots. In both lean and obese rats, as a general trend, cold exposure increased ATGL mRNA and protein levels in the different adipose depots, except in the brown adipose tissue of lean animals, where a decrease was observed. In lean rats, cold exposure strongly improved fasting up-regulation of ATGL expression in all the adipose depots. Moreover, in response to fasting, in cold-exposed lean rats, there was a stronger positive correlation between circulating nonesterified fatty acids (NEFA) and ATGL mRNA levels in the adipose depots and a higher percentage increase of circulating NEFA in comparison with control animals not exposed to cold. In obese rats, fasting-induced up-regulation of ATGL was impaired and was not improved by cold. The effects of obesity and cold exposure on HSL mRNA expression were similar to those observed for ATGL, suggesting common regulatory mechanisms for both proteins. Thus, cold exposure increases ATGL expression and improves its fasting-up-regulation in adipose tissue of lean rats. In obese rats, cold exposure also increases ATGL expression but fails to improve its regulation by fasting, which could contribute to the increased difficulty for mobilizing lipids in these animals. Copyright © 2012 Elsevier Inc. All rights reserved.

  7. Lipases and lipid droplet-associated protein expression in subcutaneous white adipose tissue of cachectic patients with cancer.

    Science.gov (United States)

    Silvério, Renata; Lira, Fábio S; Oyama, Lila M; Oller do Nascimento, Cláudia M; Otoch, José P; Alcântara, Paulo S M; Batista, Miguel L; Seelaender, Marília

    2017-08-22

    Cancer cachexia is a multifactorial metabolic syndrome characterized by marked loss of adipose tissue and skeletal muscle. Fat loss from adipose tissue in cancer cachexia is partly the result of increased lipolysis. Despite the growing amount of studies focused on elucidating the mechanisms through which lipolysis-related proteins regulate the lipolytic process, there are scarce data concerning that profile in the adipose tissue of cancer cachectic patients. Considering its fundamental importance, it was our main purpose to characterize the expression of the lipolysis-related proteins in the white adipose tissue of cachectic cancer patients. Patients from the University Hospital were divided into three groups: control, cancer cachexia (CC), and weight-stable cancer patients (WSC). To gain greater insight into adipose tissue wasting during cancer cachexia progression, we have also analyzed an experimental model of cachexia (Walker 256 carcinosarcoma). Animals were divided into: control, intermediate cachexia (IC) and terminal cachexia (TC). Subcutaneous white adipose tissue of patients and epidydimal white adipose tissue of animals were investigated regarding molecular aspects by determining the protein content and gene expression of hormone-sensitive lipase (HSL), adipose triglyceride lipase (ATGL), comparative gene identification-58 (CGI-58), perilipin 1, leptin, adiponectin, visfatin, and tumour necrosis factor alpha (TNF-alpha). We found augmented lipolysis in CC associated with increased HSL expression, as well as upregulation of ATGL expression and reduction in perilipin 1 content. In IC, there was an imbalance in the secretion of pro- and anti-inflammatory factors. The alterations at the end-stage of cachexia were even more profound, and there was a reduction in the expression of almost all proteins analyzed in the animals. Our findings show that cachexia induces important morphological, molecular, and humoral alterations in the white adipose tissue, which

  8. Nieuwe methode maakt HSL-Oost rendabel

    NARCIS (Netherlands)

    H.D. Waveren; L. Bertolini

    2011-01-01

    Een hogesnelheidslijn van Amsterdam via Utrecht en Arnhem naar de Duitse grens kost te veel en levert te weinig op, was het oordeel in 2001. Een nieuwe ontwerpmethode kijkt breder en houdt de aanpassingen in de infrastructuur beperkt. Zo wordt de HSL-Oost opeens wel rendabel.

  9. Effects of rosiglitazone and high fat diet on lipase/esterase expression in adipose tissue.

    Science.gov (United States)

    Shen, Wen-Jun; Patel, Shailja; Yu, Zaixin; Jue, Dyron; Kraemer, Fredric B

    2007-02-01

    A number of intracellular lipase/esterase have been reported in adipose tissue either by functional assays of activity or through proteomic analysis. In the current work, we have studied the relative expression level of 12 members of the lipase/esterase family that are found in white adipose tissue. We found that the relative mRNA levels of ATGL and HSL are the most abundant, being 2-3 fold greater than TGH or ADPN; whereas other intracellular neutral lipase/esterases were expressed at substantially lower levels. High fat feeding did not alter the mRNA expression levels of most lipase/esterases, but did reduce CGI-58 and WBSCR21. Likewise, rosiglitazone treatment did not alter the mRNA expression levels of most lipase/esterases, but did increase ATGL, TGH, CGI-58 and WBSCR21, while reducing ADPN. WAT from HSL-/- mice showed no compensatory increase in any lipase/esterases, rather mRNA levels of most lipase/esterases were reduced. In contrast, BAT from HSL-/- mice showed an increase in ATGL expression, as well as a decrease in ES-1, APEH and WBSCR21. Analysis of the immunoreactive protein levels of some of the lipases confirmed the results seen with mRNA. In conclusion, these data highlight the complexity of the regulation of the expression of intracellular neutral lipase/esterases involved in lipolysis.

  10. Geluidreductie van de pilotmaatregelen aan het HSL spoor in Lansingerland

    NARCIS (Netherlands)

    Dittrich, M.G.; Eisses, A.; Staats, F.; Prevo, R.; Hof, J. van 't

    2014-01-01

    TNO heeft in opdracht van ProRail geluid- en trillingsmetingen uitgevoerd aan de HSL in Lansingerland, ter bepaling van het geluidreducerend effect van raildempers en absorberende platen die in 2013 zijn geïnstalleerd op het Rhedaspoor. De HSL ter plaatse rijdt in een verdiept spoor gedeeltelijk voo

  11. Redefining Hormone Sensitive Disease in Advanced Prostate Cancer

    Directory of Open Access Journals (Sweden)

    Xiaoyu Hou

    2012-01-01

    Full Text Available Prostate cancer is the most common cancer among men in the United States. For decades, the cornerstone of medical treatment for advanced prostate cancer has been hormonal therapy, intended to lower testosterone levels, known as Androgen Deprivation Therapy (ADT. The development of hormone-resistant prostate cancer (now termed castration-resistant prostate cancer:CRPC remains the key roadblock in successful long-term management of prostate cancer. New advancements in medical therapy for prostate cancer have added to the hormonal therapy armamentarium. These new therapeutic agents not only provide a survival benefit but also show potential for reversing hormonal resistance in metastatic CRPC, and thus redefining hormonally sensitive disease.

  12. New HSL and HSV color spaces and applications

    Science.gov (United States)

    Marcu, Gabriel G.; Abe, Satoshi

    1997-02-01

    The color classification applied for large documents requires simple color transformations that enables the implementation of high speed algorithms with reasonable performance. The use of HSV and HSL color spaces in such applications is limited due to their perceptual non- linearity, even if these spaces are attractive due to the simplicity of their definition and transformation. This paper proposes new relationships of the HSL and HSV color spaces based on the replacement of the lightness and brightness definitions, with new relationships based on more perceptual lightness and brightness. The new HSL and HSV color spaces are visualized in three dimensions and the shapes are compared with the old regulate spaces. These spaces remain device dependent as the conventional HSV and HSL spaces, but the distribution of color is more suitable for some application like color clusterization. The new defined color spaces are asymmetric. It is shown how the new HSV and HSL color spaces simplify and improve the accuracy of a clusterization process. The results of clusterization process are compared in the processed image area as well as in the 3D histogram diagram. The color spaces are used for clusterization of colormaps, with application to automatic color classification in large documents (maps, blueprint documents, etc.). Results of clusterization process for blueprint documents are provided for the new introduced HSL and HSV color spaces.

  13. Insights into the molecular evolution of HslU ATPase through biochemical and mutational analyses.

    Directory of Open Access Journals (Sweden)

    Kwang Hoon Sung

    Full Text Available The ATP-dependent HslVU complexes are found in all three biological kingdoms. A single HslV protease exists in each species of prokaryotes, archaea, and eukaryotes, but two HslUs (HslU1 and HslU2 are present in the mitochondria of eukaryotes. Previously, a tyrosine residue at the C-terminal tail of HslU2 has been identified as a key determinant of HslV activation in Trypanosoma brucei and a phenylalanine at the equivalent position to E. coli HslU is found in T. brucei HslU1. Unexpectedly, we found that an F441Y mutation in HslU enhanced the peptidase and caseinolytic activity of HslV in E. coli but it showed partially reduced ATPase and SulA degradation activity. Previously, only the C-terminal tail of HslU has been the focus of HslV activation studies. However, the Pro315 residue interacting with Phe441 in free HslU has also been found to be critical for HslV activation. Hence, our current biochemical analyses explore the importance of the loop region just before Pro315 for HslVU complex functionality. The proline and phenylalanine pair in prokaryotic HslU was replaced with the threonine and tyrosine pair from the functional eukaryotic HslU2. Sequence comparisons between multiple HslUs from three different biological kingdoms in combination with biochemical analysis of E. coli mutants have uncovered important new insights into the molecular evolutionary pathway of HslU.

  14. Bacterial lipases

    NARCIS (Netherlands)

    Jaeger, Karl-Erich; Ransac, Stéphane; Dijkstra, Bauke W.; Colson, Charles; Heuvel, Margreet van; Misset, Onno

    1994-01-01

    Many different bacterial species produce lipases which hydrolyze esters of glycerol with preferably long-chain fatty acids. They act at the interface generated by a hydrophobic lipid substrate in a hydrophilic aqueous medium. A characteristic property of lipases is called interfacial activation, mea

  15. Lipase Test

    Science.gov (United States)

    ... with pancreatic duct obstruction, pancreatic cancer , and other pancreatic diseases as well as with gallbladder inflammation or kidney ... damage to the lipase-producing cells in the pancreas. This can occur in chronic diseases that affect the pancreas such as cystic fibrosis . ^ ...

  16. Human skeletal muscle perilipin 2 and 3 expression varies with insulin sensitivity

    DEFF Research Database (Denmark)

    Vigelsø Hansen, Andreas; Prats Gavalda, Clara; Ploug, Thorkil

    2013-01-01

    was obtained from vastus lateralis, and a two-step sequential euglycaemic-hy- perinsulinaemic clamp was performed. Muscle sam- ples were analyzed by Western blot for expression of perilipin 2, 3, 5, adipose triglyceride lipase (ATGL), hormone-sensitive lipase (HSL), endothelial lipase (EL) and lipoprotein...

  17. Mechanism of azithromycin inhibition of HSL synthesis in Pseudomonas aeruginosa.

    Science.gov (United States)

    Zeng, Jianming; Zhang, Ni; Huang, Bin; Cai, Renxin; Wu, Binning; E, Shunmei; Fang, Chengcai; Chen, Cha

    2016-04-14

    Pseudomonas aeruginosa is an opportunistic pathogen and a leading cause of nosocomial infections. Unfortunately, P. aeruginosa has low antibiotic susceptibility due to several chromosomally encoded antibiotic resistance genes. Hence, we carried out mechanistic studies to determine how azithromycin affects quorum sensing and virulence in P. aeruginosa. lasI and rhlI single and double mutants were constructed. We then undertook a quantitative approach to determine the optimal concentration of azithromycin and culture time that can affect the expression of HSLs. Furthermore, based on the above results, the effect on quorum sensing was analyzed at a transcriptional level. It was found that 2 μg/mL azithromycin caused a 79% decrease in 3-oxo-C12-HSL secretion during cultivation, while C4-HSL secretion was strongly repressed in the early stages. Azithromycin acts on ribosomes; to determine whether this can elicit alternative modes of gene expression, transcriptional regulation of representative virulence genes was analyzed. We propose a new relationship for lasI and rhlI: lasI acts as a cell density sensor, and rhlI functions as a fine-tuning mechanism for coordination between different quorum sensing systems.

  18. Boundary emphasis transfer function generation based on HSL color space

    Science.gov (United States)

    Li, Xiao; Wu, Jianhuang; Luo, Shengzhou; Ma, Xin

    2011-10-01

    Direct volume rendering has been received much attention since it need not to extract geometric primitives for visualization and its performance is generally better than surface rendering. Transfer functions, which are used for mapping scalar field to optical properties, are of vital importance in obtaining a sensible rendering result from volume data. Though traditional color transfer functions are in RGB color space, HSL color space that conveys semantic meanings is more intuitive and user-friendly. In this paper, we present a novel approach aims to emphasize and distinguish strong boundaries between different materials. We achieve it by using data value, gradient magnitude and dimension of the volumetric data to set opacity. Then, through a linear map from data value, gradient magnitude and second derivative to hue, saturation and lightness respectively, a color transfer function is obtained in HSL color space. Experimental tests on real-world datasets indicate that our method could achieve desirable rendering results with revealing important boundaries between different structures and indicating data value's distribution in the volume by using different colors.

  19. HSL Attenuates the Follicular Oxidative Stress and Enhances the Hair Growth in ob/ob Mice

    Directory of Open Access Journals (Sweden)

    Takeo Minematsu, PhD

    2013-10-01

    Full Text Available Summary: We demonstrated enhanced hair regeneration following topical administration of N-(3-oxododecanoyl-L-homoserine lactone (HSL in ob/ob mice. The ob/ob mice showed delayed hair regeneration (more than 6 wk after depilation, which rapidly induced transition to anagen in the hair cycle in wild-type mice. Vehicle and HSL solutions were applied to the depilated dorsal skin of ob/ob mice. The depilated skin of the HSL-treated mice was fully covered with hair, whereas no macroscopic alteration was observed in vehicle-treated group by the fourth week after depilation. Oxidative stress was drastically decreased and the expression of the antioxidative enzymes PON1 and PON3 was increased in the HSL-treated skin with highly proliferative anagen follicles. These results suggest that HSL is a candidate therapeutic agent for alopecia in metabolic syndrome.

  20. Promotion of acute-phase skin wound healing by Pseudomonas aeruginosa C4 -HSL.

    Science.gov (United States)

    Kanno, Emi; Kawakami, Kazuyoshi; Miyairi, Shinichi; Tanno, Hiromasa; Suzuki, Aiko; Kamimatsuno, Rina; Takagi, Naoyuki; Miyasaka, Tomomitsu; Ishii, Keiko; Gotoh, Naomasa; Maruyama, Ryoko; Tachi, Masahiro

    2016-12-01

    A Pseudomonas aeruginosa quorum-sensing system, which produces N-(3-oxododecanoyl)-l-homoserine lactone (3-oxo-C12 -HSL) and N-butanoyl-l-homoserine lactone (C4 -HSL), regulates the virulence factors. In our previous study, 3-oxo-C12 -HSL, encoded by lasI gene, was shown to promote wound healing. However, the effect of C4 -HSL, encoded by rhlI gene, remains to be elucidated. We addressed the effect of C4 -HSL on wounds in P. aeruginosa infection. Wounds were created on the backs of Sprague-Dawley SD rats, and P. aeruginosa PAO1 (PAO1) or its rhlI deletion mutant (ΔrhlI) or lasI deletion mutant (ΔlasI) was inoculated onto the wound. Rats were injected intraperitoneally with anti-C4 -HSL antiserum or treated with C4 -HSL at the wound surface. PAO1 inoculation led to significant acceleration of wound healing, which was associated with neutrophil infiltration and TNF-α synthesis. These responses were reversed, except for TNF-α production, when ΔrhlI was inoculated instead of PAO1 or when rats were co-treated with PAO1 and anti-C4 -HSL antiserum. In contrast, the healing process and neutrophil infiltration, but not TNF-α synthesis, were accelerated when C4 -HSL was administered in the absence of PAO1. This acceleration was not affected by anti-TNF-α antibody. These results suggest that C4 -HSL may be involved in the acceleration of acute wound healing in P. aeruginosa infection by modifying the neutrophilic inflammation. © 2015 Medicalhelplines.com Inc and John Wiley & Sons Ltd.

  1. Recovery of hormone sensitivity after salvage brachytherapy for hormone refractory localized prostate cancer

    OpenAIRE

    Dan Smith; P. Nick Plowman

    2010-01-01

    PURPOSE: Recent work has demonstrated the return of hormone sensitivity after palliative chemotherapy in androgen independent prostate cancer. We wished to establish whether a similar phenomenon existed in patients with no exposure to chemotherapy. MATERIALS AND METHODS: A review of “hormone resistant” patients who had received salvage brachytherapy for localized prostate cancer after previous external beam radiotherapy was undertaken. Three patients with subsequent biochemical re...

  2. Recovery of hormone sensitivity after salvage brachytherapy for hormone refractory localized prostate cancer

    Directory of Open Access Journals (Sweden)

    Dan Smith

    2010-06-01

    Full Text Available PURPOSE: Recent work has demonstrated the return of hormone sensitivity after palliative chemotherapy in androgen independent prostate cancer. We wished to establish whether a similar phenomenon existed in patients with no exposure to chemotherapy. MATERIALS AND METHODS: A review of “hormone resistant” patients who had received salvage brachytherapy for localized prostate cancer after previous external beam radiotherapy was undertaken. Three patients with subsequent biochemical relapse responded to the rechallenge with hormonal treatment. RESULTS: The series of patients presented here demonstrates this phenomenon occurs after salvage brachytherapy with no exposure to chemotherapy. Recovery of sensitivity is demonstrated both to androgen deprivation and to androgen receptor antagonism. The recovery of hormone sensitivity was surprisingly durable, ranging from eight months to over twenty-one months. CONCLUSIONS: Hormone sensitivity may be recovered after salvage brachytherapy. Potential mechanisms underlying these observations are discussed and the likely central role of the activity of the androgen receptor highlighted. The relevance of these findings to the management of advanced prostate cancer is considered including thoughts on the practice of intermittent anti-androgen therapy.

  3. Familial lipoprotein lipase deficiency

    Science.gov (United States)

    ... page: //medlineplus.gov/ency/article/000408.htm Familial lipoprotein lipase deficiency To use the sharing features on this page, please enable JavaScript. Familial lipoprotein lipase deficiency is a group of rare genetic ...

  4. Acid Lipase Disease

    Science.gov (United States)

    ... Page You are here Home » Disorders » All Disorders Acid Lipase Disease Information Page Acid Lipase Disease Information Page What research is being ... research to understand lipid storage diseases such as acid lipase deficiency. Additional research studies hope to identify ...

  5. A simple, rapid, sensitive method detecting homoserine lactone (HSL)-related compounds in microbial extracts.

    Science.gov (United States)

    Singh, Maya Prakash; Greenstein, Michael

    2006-04-01

    A simple, rapid, sensitive microtiter plate method detecting N-acyl homoserine lactone (HSL)-related compounds was established using an Agrobacterium tumefaciens strain harboring a traG::lacZ/traR reporter gene responsive to HSLs. This strain did not produce its own HSL, but the traG::lacZ reporter gene was induced only when its transcription activator TraR detected a cognate exogenous HSL. Therefore, the assay was expected to be highly specific for HSL-related compounds. Induction of the reporter gene, leading to production of beta-galactosidase enzyme, was measured by using two different beta-galactosidase substrates, X-gal and Galacton-Star, for colorimetric and chemiluminometric detection, respectively. The screen was validated in both the 96-well and 384-well plate formats, and extracts derived from 696 different microbial isolates, mostly unidentified actinomycetes isolated from diverse locations, were tested. Crude extracts of 81 (11.64%) cultures tested positive for HSL-related compounds, and an additional 34 (4.8%) crude extracts showed a moderate to weak signal for HSLs. Data from the fractionated samples, however, suggested a much higher prevalence of HSL signals in these extracts. Of 144 crude extracts fractionated into 10 individual samples at a 10x concentration, 72 (50%) cultures tested positive for HSLs. Six cultures were active only in the crude extract, 18 were active both in crude and one or more of their fractions, and an additional 48 were active in just one or more of their fractions. This finding may be the first to suggest such a high prevalence of HSL-signals found in nature, and a large number of actinomycetes in our collection appeared to produce HSL-related compounds.

  6. Effects of IGF-Ⅰ and GLP-Ⅰon activity of HSL in culture bovine adipocyte in vitro%IGF-Ⅰ、GLP-Ⅰ对体外培养脂肪细胞HSL酶活性的影响

    Institute of Scientific and Technical Information of China (English)

    王娅; 邓俊良; 王哲; 左之才; 胡延春

    2013-01-01

    取体外培养单层生长良好脂肪细胞,培养介质中分别添加0、10、20、30、40、50μg/L的胰岛素样生长因子Ⅰ(IGF-Ⅰ),0、100、250、500、750、1 000 nmol/L的胰高血糖素样肽Ⅰ(GLP-Ⅰ),培养24 h后提取总蛋白,每个处理3个重复,分别采用脂肪酶测定试剂盒检测HSL酶活性的影响.结果表明,IGF-Ⅰ、GLP-Ⅰ抑制脂肪细胞内HSL酶活性,存在剂量依赖性.%A test was designed to investigate whether IGF-Ⅰ,GLP-Ⅰ affect the activity of HSL in adipose cells from a newborn Holstan calf.IGF-Ⅰ,GLP-Ⅰ were added to the media with 0、10、20、30、40、50 μg/L,0、100、250、500、750、1 000 nmol/L,respectively.Activity of HSL in adipose cells was determined by the Kit of Lipase Activity Assay.The results showed that the activity of HSL was suppressed in adipose cells treated with IGF-Ⅰ,GLP-Ⅰ respectively.This suppression was dose dependent.

  7. Pseudomonas aeruginosa auto inducer3-oxo-C12-HSL exerts bacteriostatic effect and inhibits Staphylococcus epidermidis biofilm.

    Science.gov (United States)

    Singh, Pradeep Kumar; Yadav, Vivek Kumar; Kalia, Manmohit; Dohare, Suhaga; Sharma, Deepmala; Agarwal, Vishnu

    2017-09-01

    Pseudomonas aeruginosa has evolved the 3-oxo-C12-HSL and C4-HSL based quorum sensing system which is responsible for the regulation of various virulence factors and helps to dominates over other bacterial species. Staphylococcus epidermidis has frequently been reported with P. aeruginosa while the role of C4-HSL and 3-oxo-C12-HSL on the S. epidermidis had widely been unexplored, and as per our knowledge, this is the first report on the impact of C4-HSL and 3-oxo-C12-HSL overS. epidermidis growth and biofilm. We found that among the two AHL molecules; only 3-oxo-C12-HSL was able to exert a significant effect in all the experiments including growth and biofilm of S. epidermidis. 3-oxo-C12-HSL at 100 μM and 200 μM concentrations were able to initiate the apparent transient type of planktonic growth inhibition in S. epidermidis. Microscopic analysis and biofilm quantification assay showed the inhibitory effect of 3-oxo-C12-HSL against S. epidermidis biofilm, initial attachment, and EPS production. The study concludes that P. aeruginosa associated 3-oxo-C12-HSL exerts the inhibitory effect on S. epidermidis growth and biofilms and thus it may also help Pseudomonasto dominate under the co-infection conditions. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Mitochondrial localization of the threonine peptidase PfHslV, a ClpQ ortholog in Plasmodium falciparum.

    Science.gov (United States)

    Tschan, Serena; Kreidenweiss, Andrea; Stierhof, York-Dieter; Sessler, Nicole; Fendel, Rolf; Mordmüller, Benjamin

    2010-11-01

    Plasmodium falciparum belongs to a group of eukaryotes expressing an ortholog of the prokaryotic T1-threonine peptidase, heat shock locus V (HslV). Bacterial HslV is a particularly well studied protease, due to its structural and biochemical similarity to the eukaryotic proteasome. Plasmodium falciparum HslV (PfHslV) is expressed in schizonts and merozoites of the asexual blood stage. Strong sequence conservation between plasmodial species, absence of HslV homologs in the human genome, and availability of specific inhibitors led us to explore its function and potential use as a drug target. In a first step, we investigated localization of PfHslV, using a bioinformatics approach and a transgenic P. falciparum line expressing a PfHslV-enhanced yellow fluorescent protein (EYFP) fusion protein from the endogenous pfhslV locus. PfHslV-EYFP was found in the mitochondrial matrix under fluorescence and immunoelectron microscopy. Endogenous, non-modified PfHslV was present in purified mitochondria and interference with mitochondrial membrane potential by drug treatment led to impairment of PfHslV processing. Import of heterologous EYFP into the plasmodial mitochondrion is mediated by the N-terminal 37 amino acids of PfHslV. PfHslV's targeting sequence is also functional in human cells, demonstrating strong conservation of mitochondrial targeting in eukaryotes. In conclusion, our data shows that PfHslV is located to the plasmodial mitochondrion and presumably has vital function within this organelle which makes it an attractive target for interventions.

  9. Fatty Acid Signaling: The New Function of Intracellular Lipases

    Directory of Open Access Journals (Sweden)

    Zuzana Papackova

    2015-02-01

    Full Text Available 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.

  10. Genome sequencing and systems biology analysis of a lipase-producing bacterial strain.

    Science.gov (United States)

    Li, N; Li, D D; Zhang, Y Z; Yuan, Y Z; Geng, H; Xiong, L; Liu, D L

    2016-03-18

    Lipase-producing bacteria are naturally-occurring, industrially-relevant microorganisms that produce lipases, which can be used to synthesize biodiesel from waste oils. The efficiency of lipase expression varies between various microbial strains. Therefore, strains that can produce lipases with high efficiency must be screened, and the conditions of lipase metabolism and optimization of the production process in a given environment must be thoroughly studied. A high efficiency lipase-producing strain was isolated from the sediments of Jinsha River, identified by 16S rRNA sequence analysis as Serratia marcescens, and designated as HS-L5. A schematic diagram of the genome sequence was constructed by high-throughput genome sequencing. A series of genes related to lipid degradation were identified by functional gene annotation through sequence homology analysis. A genome-scale metabolic model of HS-ML5 was constructed using systems biology techniques. The model consisted of 1722 genes and 1567 metabolic reactions. The topological graph of the genome-scale metabolic model was compared to that of conventional metabolic pathways using a visualization software and KEGG database. The basic components and boundaries of the tributyrin degradation subnetwork were determined, and its flux balance analyzed using Matlab and COBRA Toolbox to simulate the effects of different conditions on the catalytic efficiency of lipases produced by HS-ML5. We proved that the catalytic activity of microbial lipases was closely related to the carbon metabolic pathway. As production and catalytic efficiency of lipases varied greatly with the environment, the catalytic efficiency and environmental adaptability of microbial lipases can be improved by proper control of the production conditions.

  11. Role of the N terminus in enzyme activity, stability and specificity in thermophilic esterases belonging to the HSL family.

    Science.gov (United States)

    Mandrich, Luigi; Merone, Luigia; Pezzullo, Margherita; Cipolla, Laura; Nicotra, Francesco; Rossi, Mosè; Manco, Giuseppe

    2005-01-21

    A superposition between the structures of Alicyclobacillus acidocaldarius esterase 2 (EST2) and Burkholderia cepacia lipase, the latter complexed with a phosphonate inhibitor, allowed us to hypothesize for the EST2 N terminus a role in restricting the access to the active site and therefore in modulating substrate specificity. In order to test this hypothesis we generated by site-directed mutagenesis some truncated versions of EST2 and its double mutant M211S/R215L (S/L) at the N terminus. In parallel, an analysis of the Sulfolobus solfataricus P2 genome allowed us to identify a gene coding for a putative esterase of the HSL family having a natural deletion of the corresponding region. The product of this gene and the above-mentioned EST2 mutants were expressed in Escherichia coli, purified and characterised. These studies support the notion that the N terminus affects substrate specificity other than several other enzyme parameters. Although the deletions afforded a tenfold and 550-fold decrease in catalytic efficiency towards the best substrate pNP-hexanoate at 50 degrees C for EST2 and S/L, respectively, the analysis of the specific activities with different triacylglycerols with respect to pNP-hexanoate showed that their ratios were higher for deleted versus non-deleted enzymes, on all tested substrates. In particular, the above ratios for glyceryl tridecanoate were 30-fold and 14-fold higher in S/L and EST2 deleted forms, respectively, compared with their full-length versions. This behaviour was confirmed by the analysis of the S.solfataricus esterase, which showed similar specific activities on pNP-hexanoate and triacylglycerols; in addition, higher activities on the latter substrates were observed in comparison with EST2, S/L and their deleted forms. Finally, a dramatic effect on thermophilicity and thermostability in the EST2 deleted forms was observed. This is the first report highlighting the importance of the "cap" domain in the HSL family, since the N

  12. The Research on Color Grading of Green Jade Images Based on HSL Chromaticity Analysis

    Directory of Open Access Journals (Sweden)

    Zhang Lili

    2016-01-01

    Full Text Available This study aims to establish a simple and convenient method suitable for the market to grade the jade colors. Based on the HSL color model and with the assistance of the HSL analysis on green jade images acquired by camera flashlights, this study obtains the following six kinds of jade colors: melon green, emerald green, yellow green, bean green, light bean green, and light green, based on which the law governing the values and the value range can be determined, and the respective HSL value ranges ,together with their threshold values, for the seven green jades can be finally found, thus laying an important foundation for further researches on computer-based quick grading. It is also hoped that this method will be widely applied and promoted in the marketplace and realizes our ultimate goal of setting guidelines to commercial jade prices.

  13. Transplantation of rat hepatic stem-like (HSL) cells with collagen matrices.

    Science.gov (United States)

    Ueno, Yasuharu; Nagai, Hirokazu; Watanabe, Go; Ishikawa, Kiyoshi; Yoshikawa, Kiwamu; Koizumi, Yukio; Kameda, Takashi; Sugiyama, Toshihiro

    2005-12-01

    Organ restitution using somatic stem cells is of great clinical interest. Recent advances in the field of tissue engineering have demonstrated that the use of collagen matrices as scaffolds facilitates tissue reconstruction. Here, we examine the efficacy of transplantation of HSL cells, a previously established liver epithelial cell line with a potential for differentiation, using collagen scaffolds. To this end, HSL cells were transplanted into Nagase's analbuminemic rat with spongy or gelatinous type I collagen matrices. Consequently, immunohistochemical analyses and genomic PCR experiments revealed engraftment of the transplanted cells. Furthermore, the levels of serum albumin in recipient rats were found to increase up to 2.5-fold relative to controls after transplantation. These findings suggest that HSL cells are able to differentiate into functional hepatocytes in vivo, and that biodegradable collagen matrices enhance this phenomenon by providing an appropriate microenvironment for hepatocytic repopulation.

  14. Human skeletal muscle perilipin 2 and 3 expression varies with insulin sensitivity

    DEFF Research Database (Denmark)

    Vigelsø Hansen, Andreas; Prats Gavalda, Clara; Ploug, Thorkil;

    2013-01-01

    was obtained from vastus lateralis, and a two-step sequential euglycaemic-hy- perinsulinaemic clamp was performed. Muscle sam- ples were analyzed by Western blot for expression of perilipin 2, 3, 5, adipose triglyceride lipase (ATGL), hormone-sensitive lipase (HSL), endothelial lipase (EL) and lipoprotein...... lipase (LPL). Results: Perilipin 3 expression was higher in T2D compared to CON. Perilipin 2 expression was higher in CON than T2D. We observed no difference in expression of perilipin 5, ATGL, HSL, EL or LPL between the groups. Con- clusions: In the present study the muscle perilipin 3 expression...

  15. Bacterial lipases for biotechnological applications

    NARCIS (Netherlands)

    Jaeger, Karl-Erich; Schneidinger, Bernd; Rosenau, Frank; Werner, Michael; Lang, Dietmar; Dijkstra, Bauke W.; Schimossek, Klaus; Zonta, Albin; Reetz, Manfred T.

    1997-01-01

    Lipase genes originating from the Gram-negative bacteria Serrutiu marcescens and Pseudomonus urruginosa were cloned. S. marcescens lipase was overexpressed in Escherichia coli yielding inclusion bodies which were purified and finally refolded to give enzymatically active lipase. The lipase operon of

  16. De hogesnelheidstrein via Den Haag?: Kritische reactie op de Nieuwe HSL-nota

    NARCIS (Netherlands)

    Lutje Schipholt, L.R.; Schoemaker, T.J.H.; Egeter, B.; Wiggenraad, P.B.L.

    1994-01-01

    Rapport in opdracht van de Gemeente Den Haag. In deze studie is de Nieuwe HSL-nota aan een kritische toetsing onderworpen. Deze toetsing is langs drie hoofdlijnen aangepakt, te weten de toetsing van de visie op de HST in Nederland en de prognose van het aantal te verwachten reizigers, de toetsing v

  17. The Functional State of Hormone-Sensitive Adenylyl Cyclase Signaling System in Diabetes Mellitus

    Directory of Open Access Journals (Sweden)

    Alexander O. Shpakov

    2013-01-01

    Full Text Available Diabetes mellitus (DM induces a large number of diseases of the nervous, cardiovascular, and some other systems of the organism. One of the main causes of the diseases is the changes in the functional activity of hormonal signaling systems which lead to the alterations and abnormalities of the cellular processes and contribute to triggering and developing many DM complications. The key role in the control of physiological and biochemical processes belongs to the adenylyl cyclase (AC signaling system, sensitive to biogenic amines and polypeptide hormones. The review is devoted to the changes in the GPCR-G protein-AC system in the brain, heart, skeletal muscles, liver, and the adipose tissue in experimental and human DM of the types 1 and 2 and also to the role of the changes in AC signaling in the pathogenesis and etiology of DM and its complications. It is shown that the changes of the functional state of hormone-sensitive AC system are dependent to a large extent on the type and duration of DM and in experimental DM on the model of the disease. The degree of alterations and abnormalities of AC signaling pathways correlates very well with the severity of DM and its complications.

  18. 1株脂肪酶产生菌的筛选鉴定及其脂肪酶基因的克隆表达%Isolation, identification of a producing lipase strain and the lipase gene cloning and expression

    Institute of Scientific and Technical Information of China (English)

    刘义; 孟丽君; 楼梦; 姚汉超; 刘修齐; 刘小兰; 俆可瀚; 李娜; 刘德立

    2011-01-01

    A producing lipase strain was isolated from castor-oil plant soil in Wuhan sur-burb. According to morphological characteristics, physiological biochemical test and sequence analysis of 16S rDNA, in addition to the phylogenetic tree constructed, the ho-mology between strain HS-L5 and Serratia marcescens is up to 99%. Therefore, strain HS-L5 is preliminarily identified as Serratia sp. , named Serratia sp. HS-L5. Meanwhile, the lipase gene of HS-L5 was cloned and the whole gene sequence is 1 842 bp long, encoding 614 amino acids. The homology compared with HpA of Serratia marcescens SM6 is close to 98%. The amino acids squences of Serratia sp. HS-L5 contain a lipase consensus sequence-G-X-S-X-G-. The gene of HS-Lip5 was sub-cloned into expression plasmid pET-28a and expressed in E. coli. Nowadays, there are few reports about Serratia marcescens producing lipase isolated from wild environment.%从武汉市郊蓖麻地土壤中分离到1株产脂肪酶菌株,结合形态观察和生理生化鉴定,扩增16S rDNA并测序,运用Blast比对构建了进化树,发现该菌与粘质沙雷氏菌(Serratia marcescens)的同源性高达99%,初步鉴定该菌属于沙雷氏菌属(Serratia),命名为Serratia sp.HS-L5.克隆了脂肪酶基因HS-Lip5,该基因全长1 842 bp,编码氨基酸614个.序列经Blast比对分析发现与Serratiamarcescens SM6的lipA序列的同源性高达98%,经氨基酸序列初步分析,发现Serratia sp.HS-L5的氨基酸序列包含脂肪酶共有序列-G-X-S-X-G-.将HS-Lip5基因克隆到表达载体pET-28a,并在E.coli中获得了表达.

  19. 凌华推出四轴远程运动控制模块HSL-4XMO系列

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    凌华科技推出四轴远程运动控制模块HSL-4XMO-CG-N/P以及HSL-4XMO-CD-N/P-HSL-4XMO可提供四轴脉冲输出,控制伺服马达、步进马达及线性马达,具有远程控制、省配线以及单一系统多轴控制等特点,非常适合在机械自动化领域应用。

  20. Decrease in intramuscular lipid droplets and translocation of HSL in response to muscle contraction and epinephrine

    DEFF Research Database (Denmark)

    Prats, Clara; Donsmark, Morten; Qvortrup, Klaus

    2006-01-01

    A better understanding of skeletal muscle lipid metabolism is needed to identify the molecular mechanisms relating intramuscular triglyceride (IMTG) to muscle metabolism and insulin sensitivity. An increasing number of proteins have been reported to be associated with intracellular triglyceride (...... show a decrease in IMTG in response to both lipolytic stimuli. Furthermore, we identify two PAT family proteins, ADRP and TIP47, associated with IMTG. Finally, we demonstrate HSL translocation to IMTG and ADRP after stimulation with epinephrine or contraction....

  1. Disrupting ER-associated protein degradation suppresses the abscission defect of a weak hae hsl2 mutant in Arabidopsis

    Science.gov (United States)

    Baer, John; Taylor, Isaiah; Walker, John C.

    2016-01-01

    In Arabidopsis thaliana, the process of abscission, or the shedding of unwanted organs, is mediated by two genes, HAESA (HAE) and HAESA-LIKE 2 (HSL2), encoding receptor-like protein kinases (RLKs). The double loss-of-function mutant hae-3 hsl2-3 is completely deficient in floral abscission, but, interestingly, the hae-3 hsl2-9 mutant displays a less severe defect. This mutant was chosen for an ethyl methanesulfonate (EMS) screen to isolate enhancer and suppressor mutants, and two such suppressors are the focus of this study. Pooled DNA from the F2 generation of a parental backcross was analyzed by genome sequencing to reveal candidate genes, two of which complement the suppressor phenotype. These genes, EMS-MUTAGENIZED BRI1 SUPPRESSOR 3 (EBS3) and EBS4, both encode mannosyltransferases involved in endoplasmic reticulum (ER)-associated degradation (ERAD) of proteins. Further analysis of these suppressor lines revealed that suppressor mutations are acting solely on the partially functional hsl2-9 mutant receptor to modify the abscission phenotype. Expressing a hsl2-9–yellow fluorescent protein (YFP) transgene in ebs3 mutants yields a higher fluorescent signal than in EBS3/ebs3, suggesting that these mutants restore abscission by disrupting ERAD to allow accumulation of the hsl2-9 receptor, which probably escapes degradation to be trafficked to the plasma membrane to regain signaling. PMID:27566817

  2. MicroRNA: sex steroids, hormonal carcinogenesis, hormonal sensitivity of tumor tissue

    Directory of Open Access Journals (Sweden)

    A. V. Malek

    2015-01-01

    Full Text Available Sex hormones, regulating normal physiological processes of most tissues and organs, are considered to be one of the key factors in the development and progression of the reproductive system cancer. Recently, the importance of the system for post-transcriptional control of gene expression mediated by short single-stranded RNA molecules (microRNA became evident. This system is involved in regulation of normal physiological processes and in the pathogenesis of many diseases, including cancer. In review we discuss the relationship between the two regulatory systems – sex hormones and microRNAs. The relationship of these systems is considered in the context of two tumors – breast and prostate cancer. In particular, the history of research on the role of sex hormones in the pathogenesis of breast cancer and prostate cancer is briefly covered. Additionally, modern scientific data on the biogenesis and biological role of microRNAs are presented in more detail. In the cells of the hormone-sensitive tissues, sex hormones regulate the microRNA-mediated machinery of gene expression control by two known ways: specifically, affecting the activity of individual microRNA molecules and non-specifically by altering the efficiency of microRNA biogenesis and activity of RNA-induced silencing complex. This downstream regulatory network substantially enhances biological effects of sex hormones at physiological conditions. Malignant transformation leads to a distortion of the regulatory effects of sex hormones that crucially influence the system of microRNA-regulated post-transcriptional control of gene expression. The most established and clinically significant example of such phenomenon is the loss of sensitivity of cells to the regulatory action of these hormones. As a consequence, cancer cells acquire the ability to active proliferation without stimulation with sex hormones. This effect is partly mediated by microRNAs. Also, relevant experimental data

  3. Lipase Induction in Mucor hiemalis.

    Science.gov (United States)

    Akhtar, M W; Mirza, A Q; Chughtai, M I

    1980-08-01

    The influence on lipase induction in Mucor hiemalis of different types of triglycerides containing mainly oleic acid (olive oil), erucic acid (mustard oil), or saturated fatty acids of 8 to 16 carbons (coconut oil) was studied. The fungus was grown in shake flasks in a fermentation medium containing peptone, minerals, and glucose or one of the oils as the carbon source. Maximum lipase was produced when the initial pH of the fermentation medium was kept at 4.0. Addition of Ca to the medium did not increase lipase production. The optimum pH for activity of both the mycelial and extracellular lipases was found to be 7.0. The fungus produced a significant amount of lipase in the presence of glucose, but the lipase activity increased markedly when olive oil was added to the medium at the beginning of the fermentation. Addition of olive oil at a later stage did not induce as much enzyme. Studies with washed mycelia showed that a greater amount of lipase was released when olive oil was present than when glucose was present. Among the various types of triglycerides used as the carbon source, olive oil was found to be most effective in inducing the lipase. Olive oil and mustard oil fatty acids inhibited the lipase more than those of coconut oil. The lipase induced by a particular type of triglyceride did not seem to be specific for the same triglyceride, nor was it inhibited specifically by it. Irrespective of the triglyceride used in the fermentation medium, the lipase produced was most active against coconut oil triglyceride, and this specificity, as shown by lipase activities in an n-heptane system, was not found to be due to a better emulsification of this oil. The lipase of M. hiemalis can be considered to be both constitutive and inducible.

  4. Plant lipases: partial purification of Carica papaya lipase.

    Science.gov (United States)

    Rivera, Ivanna; Mateos-Díaz, Juan Carlos; Sandoval, Georgina

    2012-01-01

    Lipases from plants have very interesting features for application in different fields. This chapter provides an overview on some of the most important aspects of plant lipases, such as sources, applications, physiological functions, and specificities. Lipases from laticifers and particularly Carica papaya lipase (CPL) have emerged as a versatile autoimmobilized biocatalyst. However, to get a better understanding of CPL biocatalytic properties, the isolation and purification of individual C. papaya lipolytic enzymes become necessary. In this chapter, a practical protocol for partial purification of the latex-associated lipolytic activity from C. papaya is given.

  5. Regulation of Lipolysis and Adipose Tissue Signaling during Acute Endotoxin-Induced Inflammation

    DEFF Research Database (Denmark)

    Rittig, Nikolaj; Bach, Ermina; Thomsen, Henrik Holm;

    2016-01-01

    = 0.001) compared with Placebo. These effects were associated with increased phosphorylation of hormone sensitive lipase (pHSL) at ser650 in adipose tissue (p = 0.03), a trend towards elevated pHSL at ser552 (p = 0.09) and cAMP-dependent protein kinase A (PKA) phosphorylation of perilipin 1 (PLIN1) (p...... lipolysis in adipose tissue and is associated with increased pHSL and signs of increased PLIN1 phosphorylation combined with a trend toward decreased insulin signaling. The combination of these mechanisms appear to be the driving forces behind the increased lipolysis observed in the early stages of acute...

  6. Identification of a bacterial-like HslVU protease in the mitochondria of Trypanosoma brucei and its role in mitochondrial DNA replication.

    Directory of Open Access Journals (Sweden)

    Ziyin Li

    2008-04-01

    Full Text Available ATP-dependent protease complexes are present in all living organisms, including the 26S proteasome in eukaryotes, Archaea, and Actinomycetales, and the HslVU protease in eubacteria. The structure of HslVU protease resembles that of the 26S proteasome, and the simultaneous presence of both proteases in one organism was deemed unlikely. However, HslVU homologs have been identified recently in some primordial eukaryotes, though their potential function remains elusive. We characterized the HslVU homolog from Trypanosoma brucei, a eukaryotic protozoan parasite and the causative agent of human sleeping sickness. TbHslVU has ATP-dependent peptidase activity and, like its bacterial counterpart, has essential lysine and N-terminal threonines in the catalytic subunit. By epitope tagging, TbHslVU localizes to mitochondria and is associated with the mitochondrial genome, kinetoplast DNA (kDNA. RNAi of TbHslVU dramatically affects the kDNA by causing over-replication of the minicircle DNA. This leads to defects in kDNA segregation and, subsequently, to continuous network growth to an enormous size. Multiple discrete foci of nicked/gapped minicircles are formed on the periphery of kDNA disc, suggesting a failure in repairing the gaps in the minicircles for kDNA segregation. TbHslVU is a eubacterial protease identified in the mitochondria of a eukaryote. It has a novel function in regulating mitochondrial DNA replication that has never been observed in other organisms.

  7. Physiological regulation of lipoprotein lipase

    NARCIS (Netherlands)

    Kersten, A.H.

    2014-01-01

    The enzyme lipoprotein lipase (LPL), originally identified as the clearing factor lipase, hydrolyzes triglycerides present in the triglyceride-rich lipoproteins VLDL and chylomicrons. LPL is primarily expressed in tissues that oxidize or store fatty acids in large quantities such as the heart, skele

  8. Geluidimmissie van Thalys en Ansaldo V250 treinen op het HSL spoor bij twee woningen in Kaag en Braassem

    NARCIS (Netherlands)

    Hof, J. van 't; Staats, F.; Graafland, F.; Prevo, R.

    2014-01-01

    TNO heeft in opdracht van ProRail de geluidimmissieniveaus gemeten van de Ansaldo V250 en de Thalys hogesnelheidstreinen die op het spoor van de HSL-Zuid met hoge snelheid rijden, bij twee woningen in de gemeente Kaag en Braassem. Tijdens dit onderzoek rijden er treinen van het type Ansaldo V250, Th

  9. Simplified assays of lipolysis enzymes for drug discovery and specificity assessment of known inhibitors.

    Science.gov (United States)

    Iglesias, Jose; Lamontagne, Julien; Erb, Heidi; Gezzar, Sari; Zhao, Shangang; Joly, Erik; Truong, Vouy Linh; Skorey, Kathryn; Crane, Sheldon; Madiraju, S R Murthy; Prentki, Marc

    2016-01-01

    Lipids are used as cellular building blocks and condensed energy stores and also act as signaling molecules. The glycerolipid/ fatty acid cycle, encompassing lipolysis and lipogenesis, generates many lipid signals. Reliable procedures are not available for measuring activities of several lipolytic enzymes for the purposes of drug screening, and this resulted in questionable selectivity of various known lipase inhibitors. We now describe simple assays for lipolytic enzymes, including adipose triglyceride lipase (ATGL), hormone sensitive lipase (HSL), sn-1-diacylglycerol lipase (DAGL), monoacylglycerol lipase, α/β-hydrolase domain 6, and carboxylesterase 1 (CES1) using recombinant human and mouse enzymes either in cell extracts or using purified enzymes. We observed that many of the reported inhibitors lack specificity. Thus, Cay10499 (HSL inhibitor) and RHC20867 (DAGL inhibitor) also inhibit other lipases. Marked differences in the inhibitor sensitivities of human ATGL and HSL compared with the corresponding mouse enzymes was noticed. Thus, ATGListatin inhibited mouse ATGL but not human ATGL, and the HSL inhibitors WWL11 and Compound 13f were effective against mouse enzyme but much less potent against human enzyme. Many of these lipase inhibitors also inhibited human CES1. Results describe reliable assays for measuring lipase activities that are amenable for drug screening and also caution about the specificity of the many earlier described lipase inhibitors.

  10. Jueming prescription and its ingredients, semen cassiae and Rhizoma Curcumae Longae, stimulate lipolysis and enhance the phosphorylation of hormone‑sensitive lipase in cultured rat white adipose tissue.

    Science.gov (United States)

    Zhang, Yue; Li, Jiaojiao; Wen, Xiuying

    2017-08-22

    The present study aimed to investigate the effect of jueming prescription (JMP) and its ingredients, semen cassiae (SC) and Rhizoma Curcumae Longae (RCL), on lipolysis, and to examine their effect on the phosphorylation of hormone‑sensitive lipase (HSL) in cultured rat white adipose tissue (WAT). Retroperitoneal WAT was aseptically excised from adult male Sprague‑Dawley rats, minced into uniform sections and subjected to ex vivo culture for 24 h. The tissue sections were then distributed into a 24‑well culture plate and treated with normal saline (vehicle), isoproterenol (ISO), JMP, SC and RCL. Non‑esterified fatty acid (NEFA) and glycerol release from the intact WAT explants were determined as a measurement of lipolysis, which were measured using NEFA and glycerol assay kits. The phosphorylation of HSL at Ser563 (P‑HSL S563) and 660 residues (P‑HSL S660) were determined using western blot analysis. The size of the adipocytes was visualized using hematoxylin and eosin (H&E) staining. It was found that JMP‑, SC‑ and RCL‑stimulated lipolysis was responsible for increasing the release of NEFAs and glycerol from the intact WAT in vitro. In addition, JMP, SC and RCL increased the levels of P‑HSL Ser563: JMP water (JW) extract, 3.52‑fold; JMP ethanol (JE) extract, 3.38‑fold; SC water (SW) extract, 4.60‑fold; SC ethanol (SE) extract, 4.20‑fold; RCL water (RW) extract, 6.98‑fold; RCL ethanol (RE) extract, 6.60‑fold. JMP, SC and RCL also increased the levels of P‑HSL Ser660: JW extract, 3.16‑fold; JE extract, 2.92‑fold; SW extract, 4.57‑fold; SE extract, 4.13‑fold; RW extract, 5.41‑fold; RE 4.96‑fold) in the WAT. The RW extract had the most marked effect. The HE staining revealed that JMP, SC and RCL reduced the size of adipocytes in the WAT. In conclusion, JMP and its ingredients, SC and RC, stimulated lipolysis and reduced the size of adipocytes, possibly via the phosphorylation of HSL in cultured rat WAT.

  11. Conservation of the abscission signaling peptide IDA during Angiosperm evolution: withstanding genome duplications and gain and loss of the receptors HAE/HSL2

    Directory of Open Access Journals (Sweden)

    Ida M. Stø

    2015-10-01

    Full Text Available The peptide INFLORESCENCE DEFICIENT IN ABSCISSION (IDA, which signals through the leucine-rich repeat receptor-like kinases HAESA (HAE and HAESA-LIKE2 (HSL2, controls different cell separation events in Arabidopsis thaliana. We hypothesize the involvement of this signaling module in abscission processes in other plant species even though they may shed other organs than A. thaliana. As the first step towards testing this hypothesis from an evolutionarily perspective we have identified genes encoding putative orthologues of IDA and its receptors by BLAST searches of publically available protein, nucleotide and genome databases for angiosperms. Genes encoding IDA or IDA-LIKE (IDL peptides and HSL proteins were found in all investigated species, which were selected as to represent each angiosperm order with available genomic sequences. The 12 amino acids representing the bioactive peptide in A. thaliana have virtually been unchanged throughout the evolution of the angiosperms; however, the number of IDL and HSL genes varies between different orders and species. The phylogenetic analyses suggest that IDA, HSL2 and the related HSL1 gene, were present in the species that gave rise to the angiosperms. HAE has arisen from HSL1 after a genome duplication that took place after the monocot - eudicots split. HSL1 has also independently been duplicated in the monocots, while HSL2 has been lost in gingers (Zingiberales and grasses (Poales. IDA has been duplicated in eudicots to give rise to functionally divergent IDL peptides. We postulate that the high number of IDL homologs present in the core eudicots is a result of multiple whole genome duplications. We substantiate the involvement of IDA and HAE/HSL2 homologs in abscission by providing gene expression data of different organ separation events from various species.

  12. Biodiesel production with immobilized lipase: A review.

    Science.gov (United States)

    Tan, Tianwei; Lu, Jike; Nie, Kaili; Deng, Li; Wang, Fang

    2010-01-01

    Fatty acid alkyl esters, also called biodiesel, are environmentally friendly and show great potential as an alternative liquid fuel. Biodiesel is produced by transesterification of oils or fats with chemical catalysts or lipase. Immobilized lipase as the biocatalyst draws high attention because that process is "greener". This article reviews the current status of biodiesel production with immobilized lipase, including various lipases, immobilization methods, various feedstocks, lipase inactivation caused by short chain alcohols and large scale industrialization. Adsorption is still the most widely employed method for lipase immobilization. There are two kinds of lipase used most frequently especially for large scale industrialization. One is Candida antartica lipase immobilized on acrylic resin, and the other is Candida sp. 99-125 lipase immobilized on inexpensive textile membranes. However, to further reduce the cost of biodiesel production, new immobilization techniques with higher activity and stability still need to be explored.

  13. Lipase Improvement: Goals and strategies

    OpenAIRE

    Bassegoda, Arnau; Cesarini, Silvia; Diaz, Pilar

    2012-01-01

    Lipases have received great attention as industrial biocatalysts in areas like oils and fats processing, detergents, baking, cheese making, surface cleaning, or fine chemistry . They can catalyse reactions of insoluble substrates at the lipid-water interface, preserving their catalytic activity in organic solvents. This makes of lipases powerful tools for catalysing not only hydrolysis, but also various reverse reactions such as esterification, transesterification, aminolysis, or thiotransest...

  14. Comparative analyses of lipoprotein lipase, hepatic lipase, and endothelial lipase, and their binding properties with known inhibitors.

    Directory of Open Access Journals (Sweden)

    Ziyun Wang

    Full Text Available The triglyceride lipase gene subfamily plays a central role in lipid and lipoprotein metabolism. There are three members of this subfamily: lipoprotein lipase, hepatic lipase, and endothelial lipase. Although these lipases are implicated in the pathophysiology of hyperlipidemia and atherosclerosis, their structures have not been fully solved. In the current study, we established homology models of these three lipases, and carried out analysis of their activity sites. In addition, we investigated the kinetic characteristics for the catalytic residues using a molecular dynamics simulation strategy. To elucidate the molecular interactions and determine potential key residues involved in the binding to lipase inhibitors, we analyzed the binding pockets and binding poses of known inhibitors of the three lipases. We identified the spatial consensus catalytic triad "Ser-Asp-His", a characteristic motif in all three lipases. Furthermore, we found that the spatial characteristics of the binding pockets of the lipase molecules play a key role in ligand recognition, binding poses, and affinities. To the best of our knowledge, this is the first report that systematically builds homology models of all the triglyceride lipase gene subfamily members. Our data provide novel insights into the molecular structures of lipases and their structure-function relationship, and thus provides groundwork for functional probe design towards lipase-based therapeutic inhibitors for the treatment of hyperlipidemia and atherosclerosis.

  15. Zoledronic acid increases docetaxel cytotoxicity through pMEK and Mcl-1 inhibition in a hormone-sensitive prostate carcinoma cell line

    OpenAIRE

    Silvestrini Rosella; Tesei Anna; Vannini Ivan; Ulivi Paola; Carloni Silvia; Brigliadori Giovanni; Fabbri Francesco; Amadori Dino; Zoli Wainer

    2008-01-01

    Abstract Background In prostate cancer, the identification of drug combinations that could reduce the tumor cell population and rapidly eradicate hormone-resistant cells potentially present would be a remarkable breakthrough in the treatment of this disease. Methods The study was performed on a hormone-sensitive prostate cancer cell line (LNCaP) grown in normal or hormone-deprived charcoal-stripped (c.s.) medium. Cell viability and apoptosis were assessed by SRB assay and Annexin-V/TUNEL assa...

  16. Influence and mechanism of N-(3-oxooxtanoyl)-L-homoserine lactone (C8-oxo-HSL) on biofilm behaviors at early stage

    Institute of Scientific and Technical Information of China (English)

    Siqing Xia; Lijie Zhou; Zhiqiang Zhang; Jixiang Li

    2012-01-01

    N-acyl-homoserines quenching,enzymatic quenching of bacterial quorum sensing,has recently applied to mitigate biofilm in membrane bioreactor.However,the effect of AHLs on the behavior of biofilm formation is still sparse.In this study,Pseudomonas aeruginosa biofilm was formed on ultra-filtration membrane under a series of N-(3-oxooxtanoyl)-L-homoserine lactone (Cs-oxo-HSL)concentrations.Diffusing Cs-oxo-HSL increased the growth rate of cells on biofilm where the concentration of Cs-oxo-HSL was over 10-7 g/L.The C8-oxo-HSL gradient had no observable influence on cell density and extracellular polymeric substances of biofilm with over 10-7 g/L Cs-oxo-HSL.Surprisingly,10-11-10-8 g/L of Cs-oxo-HSL had no effect on cell growth in liquid culture.The cell analysis demonstrated that the quorum sensing system might enhance the growth of neighboring cells in contact with surfaces into biofilm and may influence the structure and organization of biofilm.

  17. Influence and mechanism of N-(3-oxooxtanoyl)-L-homoserine lactone (C8-oxo-HSL) on biofilm behaviors at early stage.

    Science.gov (United States)

    Xia, Siqing; Zhou, Lijie; Zhang, Zhiqiang; Li, Jixiang

    2012-01-01

    N-acyl-homoserines quenching, enzymatic quenching of bacterial quorum sensing, has recently applied to mitigate biofilm in membrane bioreactor. However, the effect of AHLs on the behavior of biofilm formation is still sparse. In this study, Pseudomonas aeruginosa biofilm was formed on ultra-filtration membrane under a series of N-(3-oxooxtanoyl)-L-homoserine lactone (Cs-oxo-HSL) concentrations. Diffusing C8-oxo-HSL increased the growth rate of cells on biofilm where the concentration of C8-oxo-HSL was over 10(-7) g/L. The C8-oxo-HSL gradient had no observable influence on cell density and extracellular polymeric substances of biofilm with over 10(-7) g/L C8-oxo-HSL. Surprisingly, 10(-11)-10(-8) g/L of C8-oxo-HSL had no effect on cell growth in liquid culture. The cell analysis demonstrated that the quorum sensing system might enhance the growth of neighboring cells in contact with surfaces into biofilm and may influence the structure and organization of biofilm.

  18. Gastric Lipase Secretion in Children with Gastritis

    OpenAIRE

    Krystyna Sztefko; Krzysztof Fyderek; Andrzej Zając; Andrzej Wędrychowicz; Iwona Rogatko; Tomasik, Przemyslaw J

    2013-01-01

    Gastric lipase is one of the prepancreatic lipases found in some mammalian species and in humans. Our knowledge of the hormonal regulation of gastric lipase secretion in children and adolescents is still very limited. The aim of this study was to compare the activity of human gastric lipase (HGL) in gastric juice in healthy adolescents and in patients with gastritis. The adolescents were allocated to three groups: the first including patients with Helicobacter pylori gastritis (HPG; n = 10), ...

  19. Biodegradable products by lipase biocatalysis.

    Science.gov (United States)

    Linko, Y Y; Lämsä, M; Wu, X; Uosukainen, E; Seppälä, J; Linko, P

    1998-11-18

    The interest in the applications of biocatalysis in organic syntheses has rapidly increased. In this context, lipases have recently become one of the most studied groups of enzymes. We have demonstrated that lipases can be used as biocatalyst in the production of useful biodegradable compounds. A number of examples are given. 1-Butyl oleate was produced by direct esterification of butanol and oleic acid to decrease the viscosity of biodiesel in winter use. Enzymic alcoholysis of vegetable oils without additional organic solvent has been little investigated. We have shown that a mixture of 2-ethyl-1-hexyl esters can be obtained in a good yield by enzymic transesterification from rapeseed oil fatty acids for use as a solvent. Trimethylolpropane esters were also similarly synthesized as lubricants. Finally, the discovery that lipases can also catalyze ester syntheses and transesterification reactions in organic solvent systems has opened up the possibility of enzyme catalyzed production of biodegradable polyesters. In direct polyesterification of 1,4-butanediol and sebacic acid, polyesters with a mass average molar mass of the order of 56,000 g mol-1 or higher, and a maximum molar mass of about 130,000 g mol-1 were also obtained by using lipase as biocatalyst. Finally, we have demonstrated that also aromatic polyesters can be synthesized by lipase biocatalysis, a higher than 50,000 g mol-1 mass average molar mass of poly(1,6-hexanediyl isophthalate) as an example.

  20. Structure and Function of Lipase

    DEFF Research Database (Denmark)

    Skjold-Jørgensen, Jakob

    Lipases are triacylglycerol hydrolases (EC 3.1.1.3) which are able to act on water-insoluble esters, butdisplay very low activity towards water-soluble, monomeric substrates. This is ascribed to theircharacteristic activation mechanism occurring at the boundary between water and lipid, i.e. the w......Lipases are triacylglycerol hydrolases (EC 3.1.1.3) which are able to act on water-insoluble esters, butdisplay very low activity towards water-soluble, monomeric substrates. This is ascribed to theircharacteristic activation mechanism occurring at the boundary between water and lipid, i.......e. the waterlipidinterface. For Thermomyces lanuginosus lipase (TlL) and related lipases, activation of the enzymeinvolves a rearrangement of a structural domain, called the “lid”, which covers the active site inhomogenous aqueous solution. At the water-lipid interface, the lid is displaced from the active site andmoves...... towards an open conformation enabling the substrate to gain access, thus initiating catalysis.Lipases have been studied for decades and their functional features have drawn much attention withinindustrial applications since their first discovery. However, given that their molecular action takes placeat...

  1. Aspects of the regulation of liver lipase

    NARCIS (Netherlands)

    G.C. Schoonderwoerd (Kees)

    1986-01-01

    textabstractIt is evident that factors that influence the activity of liver lipase could be important because of the role of liver lipase in HDL-cholesterol metabolism. At the start of this study not much was known about the regulation of liver lipase. The activity had been found to be decreased aft

  2. Lipases and Its Application in Food Industry

    Institute of Scientific and Technical Information of China (English)

    WANG Ting; QIN Gang

    2010-01-01

    Lipases(triacylglycerol acylhydrolases,EC 3.1.1.3)occur widely in nature.It catalyze the hydrolysis and the synthesis of esters formed from glycerol and long-chain fatty acids.Lipases are commercially significant,this article discusses the source,structure,character and preparative method,the applications of lipases in food industry are discussed too.

  3. The Breast International Group 1-98 trial: big results for women with hormone-sensitive early breast cancer.

    Science.gov (United States)

    Monnier, Alain M

    2007-05-01

    As there is a risk for relapse in early breast cancer, especially at 1-3 years post surgery, the need for adjuvant therapy is clear. In terms of disease-free survival, aromatase inhibitors have emerged as superior to tamoxifen for the adjuvant treatment of hormone-sensitive breast cancer in several Phase III clinical trials. Of these trials, the Breast International Group (BIG) 1-98 trial stands out as unique in design, as it is the only trial to address whether an aromatase inhibitor is more effective as initial adjuvant therapy or as sequential therapy with an aromatase inhibitor and tamoxifen in either order and in rigor of end points and safety evaluations. When compared with tamoxifen, letrozole has been shown to significantly reduce recurrence risk in the overall population by 19% and also significantly reduced recurrence risk in the patient subgroups at increased risk: node-positive and previously chemotherapy-treated patients. Letrozole is the only aromatase inhibitor to demonstrate a significant 27% reduction in the risk of distant metastases (p = 0.001) in the clinically relevant, hormone receptor-positive population in the initial adjuvant setting. Recent results also suggest that letrozole in particular reduces the risk of distant metastases early on after initial surgery for breast cancer. This is important, as early distant metastatic events compose the majority of early recurrences and are a well-recognized predictor of breast cancer death. Letrozole has been found to be well tolerated in the initial adjuvant treatment setting, and these data have been confirmed by long-term safety data from the monotherapy analysis in the BIG 1-98 study. Thus far, the results from the BIG 1-98 trial provide clear support for the use of letrozole in the initial adjuvant treatment of breast cancer. Future studies will provide the definitive answer to questions of which initial adjuvant therapy is superior (i.e., anastrozole or letrozole) and information as to the

  4. Fasting energy homeostasis in mice with adipose deficiency of desnutrin/adipose triglyceride lipase.

    Science.gov (United States)

    Wu, Jiang Wei; Wang, Shu Pei; Casavant, Stéphanie; Moreau, Alain; Yang, Gong She; Mitchell, Grant A

    2012-05-01

    Adipose triglyceride lipase (ATGL) catalyzes the first step of lipolysis of cytoplasmic triacylglycerols in white adipose tissue (WAT) and several other organs. We created adipose-specific ATGL-deficient (ATGLAKO) mice. In these mice, in vivo lipolysis, measured as the increase of plasma nonesterified fatty acid and glycerol levels after injection of a β3-adrenergic agonist, was undetectable. In isolated ATGLAKO adipocytes, β3-adrenergic-stimulated glycerol release was 10-fold less than in controls. Under fed conditions, ATGLAKO mice had normal viability, mild obesity, low plasma nonesterified fatty acid levels, increased insulin sensitivity, and increased daytime food intake. After 5 h of fasting, ATGLAKO WAT showed phosphorylation of the major protein kinase A-mediated targets hormone-sensitive lipase and perilipin A and ATGLAKO liver showed low glycogen and triacylglycerol contents. During a 48-h fast, ATGLAKO mice developed striking and complex differences from controls: progressive reduction of oxygen consumption, high respiratory exchange ratio, consistent with reduced fatty acid availability for energy production, lethargy, hypothermia, and undiminished fat mass, but greater loss of lean mass than controls. Plasma of 48 h-fasted ATGLAKO mice had a unique pattern: low 3-hydroxybutyrate, insulin, adiponectin, and fibroblast growth factor 21 with elevated leptin and corticosterone. ATGLAKO WAT, liver, skeletal muscle, and heart showed increased levels of mRNA related to autophagy and proteolysis. In murine ATGL deficiency, adipose lipolysis is critical for fasting energy homeostasis, and fasting imposes proteolytic stress on many organs, including heart and skeletal muscle.

  5. Developmental, hormonal, and nutritional regulation of porcine adipose triglyceride lipase (ATGL).

    Science.gov (United States)

    Deiuliis, Jeffrey A; Shin, Jonghyun; Bae, Dongryeoul; Azain, Michael J; Barb, Richard; Lee, Kichoon

    2008-03-01

    Adipose triglyceride lipase (ATGL) is a newly identified lipase. We report for the first time the porcine ATGL sequence and characterize ATGL gene and protein expression in vitro and in vivo. Adult pig tissue expresses ATGL at high levels in the white adipose and muscle tissue relative to other tested tissues. We show that within the white adipose tissue ATGL is expressed at higher levels in the adipocyte than in the stromal-vascular fraction. Additionally, ATGL expression increases dramatically in the subcutaneous adipose during adipose development and maturation, as well as during in vitro adipogenesis. Peroxisome proliferator-activated receptor gamma transcript levels increased concomitant with ATGL gene expression, suggesting a possible role in the regulation of ATGL by adipogenic regulators. In vitro treatment of differentiated primary pig preadipocytes with insulin and forskolin decreased ATGL gene expression in a dose-dependent manner, suggesting ATGL transcript levels are hormone sensitive. In vivo experimentation showed that calorie-restriction in gilts resulted in increased ATGL mRNA and protein levels in subcutaneous and peri-renal fat tissues. Our data demonstrate that ATGL expression reacts to hormonal stimuli and plays a role in catecholamine-induced lipolysis in porcine adipose tissue.

  6. Lipase Induction in Mucor hiemalis

    OpenAIRE

    Akhtar, M. Waheed; Mirza, A. Q.; Chughtai, M. I. D.

    1980-01-01

    The influence on lipase induction in Mucor hiemalis of different types of triglycerides containing mainly oleic acid (olive oil), erucic acid (mustard oil), or saturated fatty acids of 8 to 16 carbons (coconut oil) was studied. The fungus was grown in shake flasks in a fermentation medium containing peptone, minerals, and glucose or one of the oils as the carbon source. Maximum lipase was produced when the initial pH of the fermentation medium was kept at 4.0. Addition of Ca2+ to the medium d...

  7. Lipases as biocatalyst for biodiesel production.

    Science.gov (United States)

    Fan, Xiaohu; Niehus, Xochitl; Sandoval, Georgina

    2012-01-01

    The global shortages of fossil fuels, significant increase in the price of crude oil, and increased environmental concerns have stimulated the rapid growth in biodiesel production. Biodiesel is generally produced through transesterification reaction catalyzed either chemically or enzymatically. Enzymatic transesterification draws high attention because that process shows certain advantages over the chemical catalysis of transesterification and it is "greener." This paper reviews the current status of biodiesel production with lipase-biocatalysis approach, including sources of lipases, kinetics, and reaction mechanism of biodiesel production using lipases, and lipase immobilization techniques. Factors affecting biodiesel production and economic feasibility of biodiesel production using lipases are also covered.

  8. Dissecting adipose tissue lipolysis: molecular regulation and implications for metabolic disease

    DEFF Research Database (Denmark)

    Nielsen, Thomas Svava; Jessen, Niels; Jørgensen, Jens Otto L;

    2014-01-01

    is tightly regulated by hormonal and nutritional factors. Under conditions of negative energy balance such as fasting and exercise, stimulation of lipolysis results in a profound increase in FFA release from adipose tissue. This response is crucial in order to provide the organism with a sufficient supply......Lipolysis is the process by which triglycerides are hydrolyzed to free fatty acids (FFA) and glycerol. In adipocytes, this is achieved by the sequential action of Adipose Triglyceride Lipase (ATGL), Hormone Sensitive Lipase (HSL) and Monoglyceride Lipase (MGL). The activity in the lipolytic pathway...... localization, protein-protein interactions, and protein stability/degradation. Here, we provide an overview of the recent advances in the field of adipose tissue lipolysis with particular focus on the molecular regulation of the two main lipases, ATGL and HSL and the intracellular and extracellular signals...

  9. Molecular and vibrational structure of the extracellular bacterial signal compound N-butyryl-homoserine lactone (C4-HSL)

    DEFF Research Database (Denmark)

    Bak, Jimmy; Spanget-Larsen, Jens

    2009-01-01

    The molecular and vibrational structure of the title compound (C4-HSL) was studied by experimental and theoretical methods. The infrared (IR) absorption spectrum was measured in the solid state and in CCl4 suspension. The observed absorption bands were compared with transitions obtained with B3LYP....../cc-pVTZ density functional theory (DFT) calculations. Two stable molecular conformations were predicted, corresponding to an endo- and an exo-conformer with similar energies. Intermolecular amide-amide hydrogen bonding in the crystal state was approximated by a simple cluster model, leading to excellent agreement...

  10. 胰岛素、胰高血糖素对体外培养的牛脂肪细胞HSL mRNA和ADPN mRNA丰度的影响%Effects of insulin and glucagons on abundance of ADPN mRNA and HSL mRNA in bovine adipocyte

    Institute of Scientific and Technical Information of China (English)

    丛立新; 张才; 车英玉; 刘国文; 王哲

    2007-01-01

    试验在单层培养的新生犊牛脂肪细胞中,分别添加胰岛素(INs)与胰高血糖素(GN),每个激素设置6个梯度3个重复,通过荧光定量PCR技术,观测2种激素对ADPN mRNA与HSL mRNA丰度的影响.结果表明,胰岛素抑制了脂肪细胞ADPN mRNA与HSL mRNA的表达,胰高血糖素促进了ADPN mRNA与HSL InRNA的表达.且ADPNmRNA与HSL mRNA存在相关性.

  11. 肥胖症脂肪组织ATGL和HSL mRNA的表达与胰岛素敏感性的关系

    Institute of Scientific and Technical Information of China (English)

    江榕; 刘小莺; 郑培烝; 陈晓春; 刘礼斌

    2010-01-01

    目的:测定肥胖症患者内脏及皮下脂肪组织三酰甘油水解酶(ATGL)和激素敏感脂肪酶(HSL)mRNA的表达,探讨其与胰岛素敏感性的相关性.方法:应用荧光实时定量PCR法检测皮下、内脏脂肪组织ATGL和HSL mRNA的表达,应用HOMA-IR评估胰岛素敏感性,分析其与人体测量指标和代谢指标的相关性.结果:①肥胖组内脏与皮下脂肪组织ATGL mRNA及HSL mRNA表达量均低于正常对照组皮下脂肪组织;②皮下及内脏脂肪组织ATGL mRNA表达量与体质量指数(BMI)呈负相关;③皮下及内脏脂肪组织HSL mRNA表达量与BMI、腰围呈负相关.结论:①肥胖症脂肪组织ATGL mRNA及HSL mRNA表达降低;②ATGL及HSL与肥胖相关;③ATGL及HSL与胰岛素抵抗可能无明显相关性.

  12. Regulation on expression of toll-like receptors on monocytes after stimulation with the 3-o-C12-HSL molecule from Pseudomonas aeruginosa.

    Science.gov (United States)

    Lu, Qi; Lin, Yujia; Yang, Xiqiang; Liu, Wei; Zhang, Xianhong; Huang, Daochao; Zhong, Haiying

    2012-10-01

    Quorum sensing (QS) is a type of cell-to-cell communication. The Pseudomonas aeruginosa QS molecule N-3-(oxododecanoyl)-L-homoserine lactone (3-o-C12-HSL) has the potential to modulate the immune system of its host. However, the mechanism of that activity is yet to be fully characterized. To be able to understand this activity, we determined whether 3-o-C12-HSL has a direct effect on the immune function and the expression of toll-like receptors (TLRs) in monocytes. Monocytes were cultured with 3-o-C12-HSL at different concentrations (0, 10, 25, 50, and 100 μmol/L) for 12 h; upon exposure to 3-o-C12-HSL, IL-12 production in monocytes was inhibited, monocyte proliferation was blocked, TLR2- and 4-mRNA expressions were reduced, and TLR5-mRNA expression was increased in a dose-dependent manner. Strikingly, 3-o-C12-HSL was able to significantly induce mRNA changes in the monocytes even at the lowest concentration (10 μmol/L, P < 0.05). Interestingly, though TLR2- and 4-protein levels were reduced, TLR5 protein expression was not changed. These findings provide a new perspective toward understanding the persistence of chronic inflammation in P. aeruginosa infections. They also suggest that TLR2, 4, and 5 may not share the same signaling pathways during monocyte activation.

  13. Lipase supplementation therapy: standards, alternatives, and perspectives.

    Science.gov (United States)

    Layer, Peter; Keller, Jutta

    2003-01-01

    Treatment of steatorrhea by lipase supplementation therapy has become more successful in the last decade due to better understanding of the physiology and pathophysiology of the digestive process. Porcine lipase has been the therapeutic standard for several decades and will continue to be the treatment of choice in pancreatic exocrine insufficiency. Modern therapeutic concepts recommend administration of 25,000-40,000 units of porcine lipase per meal using pH-sensitive pancreatin microspheres. In case of treatment failure, the dose should be increased, compliance should be checked, and other reasons for malabsorption should be excluded. Still, in most patients, lipid digestion cannot be completely normalized by current standard therapy, and future developments are needed for optimizing treatment. In this article, pathophysiologic characteristics of pancreatic exocrine insufficiency, prerequisites for use of alternative lipase sources as well as currently available lipases of nonporcine origin, and new developments are discussed. Current literature suggests that bovine lipase products present a theoretical alternative but play no major role in the western world. Fungal lipase has inferior properties compared with conventional products. Bacterial lipase products show promising potential and offer future therapeutic alternatives. Moreover, human pancreatic lipase gene transfer and application of bioengineered human gastric lipase appear on the horizon.

  14. Fucoidan from Marine Brown Algae Inhibits Lipid Accumulation

    Directory of Open Access Journals (Sweden)

    Changhyun Roh

    2011-08-01

    Full Text Available In this study, we elucidated the inhibitory effect of fucoidan from marine brown algae on the lipid accumulation in differentiated 3T3-L1 adipocytes and its mechanism. The treatment of fucoidan in a dose-dependent manner was examined on lipid inhibition in 3T3-L1 cells by using Oil Red O staining. Fucoidan showed high lipid inhibition activity at 200 µg/mL concentration (P < 0.001. Lipolytic activity in adipocytes is highly dependent on hormone sensitive lipase (HSL, which is one of the most important targets of lipolytic regulation. Here, we examined the biological response of fucoidan on the protein level of lipolysis pathway. The expressed protein levels of total hormone sensitive lipase (HSL and its activated form, phosphorylated-HSL were significantly increased at concentration of 200 µg/mL fucoidan. Furthermore, insulin-induced 2-deoxy-D-[3H] glucose uptake was decreased up to 51% in fucoidan-treated cells as compared to control. Since increase of HSL and p-HSL expression and decrease of glucose uptake into adipocytes are known to lead to stimulation of lipolysis, our results suggest that fucoidan reduces lipid accumulation by stimulating lipolysis. Therefore, these results suggest that fucoidan can be useful for the prevention or treatment of obesity due to its stimulatory lipolysis.

  15. HAZELNUT SEED LIPASE: EXTRACTION, PURIFICATION, AND CHARACTERIZATION

    OpenAIRE

    Kılıç, İsmail; Sağıroğlu, Ayten

    2012-01-01

    Interest in lipases has markedly increased to their potential industrial applications. Themost of lipases produced commercially are obtained from animal and microbial sources.Nowadays, also obtained from plant seeds such as sunflower, soybean, peanut, castor bean andhazelnut. Hazelnut is one of the most important foods in majority of the world and Turkey islargest hazelnut producer. In this study, It was aimed that Lipase from hazelnut seed identified asyomra species isolated, purified and ch...

  16. [Lipases in catalytic reactions of organic chemistry].

    Science.gov (United States)

    Bezborodov, A M; Zagustina, N A

    2014-01-01

    Aspects of enzymatic catalysis in lipase-catalyzed reactions of organic synthesis are discussed in the review. The data on modern methods of protein engineering and enzyme modification allowing a broader range of used substrates are briefly summarized. The application of lipase in the preparation of pharmaceuticals and agrochemicals containing no inactive enantiomers and in the synthesis of secondary alcohol enantiomers and optically active amides is demonstrated. The subject of lipase involvement in the C-C bond formation in the Michael reaction is discussed. Data on the enzymatic synthesis of construction materials--polyesters, siloxanes, etc.--are presented. Examples demonstrating the application of lipase enzymatic catalysis in industry are given.

  17. Crystal structure of Proteus mirabilis lipase, a novel lipase from the Proteus/psychrophilic subfamily of lipase family I.1.

    Directory of Open Access Journals (Sweden)

    Tyler P Korman

    Full Text Available Bacterial lipases from family I.1 and I.2 catalyze the hydrolysis of triacylglycerol between 25-45°C and are used extensively as biocatalysts. The lipase from Proteus mirabilis belongs to the Proteus/psychrophilic subfamily of lipase family I.1 and is a promising catalyst for biodiesel production because it can tolerate high amounts of water in the reaction. Here we present the crystal structure of the Proteus mirabilis lipase, a member of the Proteus/psychrophilic subfamily of I.1lipases. The structure of the Proteus mirabilis lipase was solved in the absence and presence of a bound phosphonate inhibitor. Unexpectedly, both the apo and inhibitor bound forms of P. mirabilis lipase were found to be in a closed conformation. The structure reveals a unique oxyanion hole and a wide active site that is solvent accessible even in the closed conformation. A distinct mechanism for Ca²⁺ coordination may explain how these lipases can fold without specific chaperones.

  18. Selectivity of lipases for estolides synthesis

    NARCIS (Netherlands)

    Todea, Anamaria; Frissen, A.E.; Otten, Linda G.; Arends, I.; Peter, F.; Boeriu, C.G.

    2015-01-01

    Lipase-catalyzed synthesis of estolides starting from different saturated (C16 16OH, C18 12OH) and unsaturated (C18:1 9 cis 12-OH) hydroxy-fatty acids was investigated. For this reason, the catalytic efficiency of several native and immobilized lipases in different organic reaction media at temperat

  19. Organic Solvent Tolerant Lipases and Applications

    Directory of Open Access Journals (Sweden)

    Shivika Sharma

    2014-01-01

    Full Text Available Lipases are a group of enzymes naturally endowed with the property of performing reactions in aqueous as well as organic solvents. The esterification reactions using lipase(s could be performed in water-restricted organic media as organic solvent(s not only improve(s the solubility of substrate and reactant in reaction mixture but also permit(s the reaction in the reverse direction, and often it is easy to recover the product in organic phase in two-phase equilibrium systems. The use of organic solvent tolerant lipase in organic media has exhibited many advantages: increased activity and stability, regiospecificity and stereoselectivity, higher solubility of substrate, ease of products recovery, and ability to shift the reaction equilibrium toward synthetic direction. Therefore the search for organic solvent tolerant enzymes has been an extensive area of research. A variety of fatty acid esters are now being produced commercially using immobilized lipase in nonaqueous solvents. This review describes the organic tolerance and industrial application of lipases. The main emphasis is to study the nature of organic solvent tolerant lipases. Also, the potential industrial applications that make lipases the biocatalysts of choice for the present and future have been presented.

  20. 21 CFR 184.1415 - Animal lipase.

    Science.gov (United States)

    2010-04-01

    ... defined in § 170.3(o)(9) of this chapter to hydrolyze fatty acid glycerides. (2) The ingredient is used in... Substances Affirmed as GRAS § 184.1415 Animal lipase. (a) Animal lipase (CAS Reg. No. 9001-62-1) is an enzyme... tissue. The enzyme preparation may be produced as a tissue preparation or as an aqueous extract. Its...

  1. Lipase in milk, curd and cheese

    NARCIS (Netherlands)

    Geurts, T.J.; Lettink, F.J.; Wouters, J.T.M.

    2003-01-01

    Presence of lipase in milk, curd, whey and cheese was studied. A small amount of the product was added to a large volume of lipase-free whole milk that had been made sensitive to lipolysis by homogenization. Increase of the acidity of the fat in the mixture, determined after incubation, was

  2. Lipase in milk, curd and cheese

    NARCIS (Netherlands)

    Geurts, T.J.; Lettink, F.J.; Wouters, J.T.M.

    2003-01-01

    Presence of lipase in milk, curd, whey and cheese was studied. A small amount of the product was added to a large volume of lipase-free whole milk that had been made sensitive to lipolysis by homogenization. Increase of the acidity of the fat in the mixture, determined after incubation, was interpre

  3. Characterization of N-butanoyl-L-homoserine lactone (C4-HSL) deficient clinical isolates of Pseudomonas aeruginosa.

    Science.gov (United States)

    Boşgelmez-Tinaz, Gülgün; Ulusoy, Seyhan

    2008-01-01

    In the opportunistic pathogen Pseudomonas aeruginosa, the production of several virulence factors such as elastase, rhamnolipids and pyocyanin depends on cell-to-cell signaling or quorum sensing (QS) involving N-acylhomoserine lactone (AHL) signal molecules. In vitro studies with laboratory strains and virulence studies in animals with these same strains have demonstrated the contribution of QS to the pathogenesis of P. aeruginosa. However, the importance of P. aeruginosa QS systems in the development of human infections is not clearly known. In order to determine if deficiency within the QS system compromises the ability of P. aeruginosa to cause infections in humans, we collected 50 P. aeruginosa clinical isolates. Phenotypic characterization showed that isolates I-457, I-458, I-459 and I-461 were defective in the production of N-butanoyl-l-homoserine lactone (C4-HSL) signaling molecule and virulence factors elastase, protease, pyocyanin and rhamnolipids. Analysis of the sequences of the lasR, lasI, rhlR and rhlI genes of these four isolates showed that two of the four isolates had mutational defects in both rhlR and rhlI genes while other two isolates were only mutated in the rhlI gene. The combination of rhlR and rhlI mutations or only rhlI mutation probably explains their C4-HSL and virulence factors deficiencies. These observations suggest that QS deficient P. aeruginosa clinical isolates are able to cause infections and that in addition to known virulence factors, factors yet unidentified may contribute to the pathogenesis of P. aeruginosa.

  4. The quorum-sensing molecule N-3-oxododecanoyl homoserine lactone (3OC12-HSL) enhances the host defence by activating human polymorphonuclear neutrophils (PMN).

    Science.gov (United States)

    Wagner, Christof; Zimmermann, Sabine; Brenner-Weiss, Gerald; Hug, Friederike; Prior, Birgit; Obst, Ursula; Hänsch, Gertrud Maria

    2007-01-01

    The P. aeruginosa quorum-sensing molecule N-3-oxododecanoyl homoserine lactone (3OC12-HSL) interacts not only with bacteria, but also with mammalian cells, among others with those of the immune defence system. We focussed on the possible interaction of 3OC12-HSL with human polymorphonuclear neutrophils (PMN), because these cells are the first to enter an infected site. We found that 3OC12-HSL attracts PMN, and up-regulates expression of receptors known to be involved in host defence, including the adhesion proteins CD11b/CD18 and the immunoglobulin receptors CD16 and CD64. Furthermore, the uptake of bacteria (phagocytosis), which is crucial for an efficient defence against infection, was enhanced. Thus, recognising and responding to 3OC12-HSL not only attracts the PMN to the site of a developing biofilm, but also reinforces their defence mechanisms, and hence could be a means to control the infection in an early stage and to prevent biofilm formation.

  5. Heterologous expression systems for lipases: a review.

    Science.gov (United States)

    Valero, Francisco

    2012-01-01

    The production of heterologous lipases is one of the most promising strategies to increase the productivity of the bioprocesses and to reduce costs, with the final objective that more industrial lipase applications could be implemented. In this chapter, an overview of the most common microbial expression systems for the overproduction of microbial lipases is presented. Prokaryotic system as Escherichia coli and eukaryotic systems as Saccharomyces cerevisiae and Pichia pastoris are analyzed and compared in terms of productivity, operational, and downstream processing facilities. Finally, an overview of heterologous Candida rugosa and Rhizopus oryzae lipases, two of the most common lipases used in biocatalysis, is presented. In both cases, P. pastoris has been shown as the most promising host system.

  6. Effect of NPY on HSL and ACC of bovine adipocyte%NPY对犊牛脂肪细胞HSL和ACC的影响

    Institute of Scientific and Technical Information of China (English)

    张才; 杨自军; 王哲; 刘国文

    2011-01-01

    试验探讨NPY在脂肪动员中的作用.体外培养犊牛脂肪细胞,通过添加不同浓度的外源性NPY(0、50、100、200、500、1 000 pg/ml)观察对脂肪细胞脂肪动员关键酶的影响.结果表明:①NPY降低脂肪细胞HSL mRNA的水平和活性,具有一定的剂量依赖性;②NPY可直接上调ACC的表达,并具一定的剂量依赖性;③随着NPY添加时间的延长,脂肪细胞HSL mRNA的水平和活性逐渐降低;④随着NPY添加时间的延长,脂肪细胞ACC的表达也逐渐升高.因此,NPY可在一定程度上促进脂肪合成,抑制脂肪动员的发生.%The focal point of this study is to probe the regulation mechanism of NPY on the above mentioned metabolism. Bovine preadipocyte was cultured by the method of adipocyte culture reported by previous literature. The effect of NPY (0,50,100,200,500,1 000 pg/ml)on mRNA expression of HSL,ACC in adipocyte were detected by fluorescent quantitation PCR. The result showed that: the expression levels of HSL mRNA in fat cell were decreased with increasing concentration of NPY in culture media. Enzyme activity of HSL was also lower than the controls. The level of ACC mRNA in fat cell was increased with increasing concentration of NPY -in culture media. The effect of 200 pg/ml NPY (0, 4, 8, 12, 16, 24 h) on the above parameters were detected by fluorescent quantitation PCR. The result showed that: the expression levels of ACC mRNA in fat cell were increased with delaying the time treated by NPY, and the activity of HSL and the expressing of HSL mRNA were decreased. It was conclued that NPY could decrease fat mobilizationin in adipocyte, and those effect of NPY were dose dependent and time dependence.

  7. Effects of IGF-Ⅰ and GLP-Ⅰon abundance of HSL mRNA of bovine adipocyte cultured in vitro%胰岛素样生长因子Ⅰ和胰高血糖素样肽Ⅰ对体外培养的脂肪细胞HSL mRNA表达的影响

    Institute of Scientific and Technical Information of China (English)

    王娅; 邓俊良; 王哲; 左之才

    2011-01-01

    为阐明胰岛素样生长因子Ⅰ(IGF-Ⅰ)、胰高血糖素样肽Ⅰ(GLP-Ⅰ)在乳牛脂肪代谢过程中的调控作用,应用荧光定量PCR法检测IGF-Ⅰ、GLP-Ⅰ对体外培养的脂肪细胞激素敏感脂肪酶(HSL)mRNA丰度的影响。结果显示,随着IGF-Ⅰ、GLP-Ⅰ添加浓度的升高,脂肪细胞HSL mRNA表达逐渐下降。结果表明,IGF-Ⅰ、GLP-Ⅰ直接调控脂肪细胞HSL mRNA的表达。%In order to investigate the regulation roles of IGF-Ⅰand GLP-Ⅰin the lipometabolism of dairy cows,the effects of them in bovine were observed by the expression level of HSL mRNA of cultured neonatal calf lipocytes using fluorescent quantitative RT-PCR.The expression level of HSL mRNA decreased following with the increasing concentrations of IGF-Ⅰand GLP-Ⅰ in the media.These results indicated that the IGF-Ⅰand GLP-Ⅰcould regulate the mRNA expressions of cultured adipose cells HSL directly.

  8. Lipase catalyzed synthesis of aromatic esters of sugar alcohols

    NARCIS (Netherlands)

    Croitoru, R.; Broek, van den L.A.M.; Frissen, A.E.; Davidescu, C.M.; Peter, F.; Boeriu, C.G.

    2011-01-01

    Commercially available lipases (Candida antarctica lipase B, Novozyme 435, Thermomyces lanuginosus lipase, and Lipozyme TL IM), as well as sol-gel immobilized lipases, have been screened for their ability to acylate regioselectively xylitol, sorbitol, and mannitol with a phenolic ester in a binary m

  9. Cloning, purification and characterisation of Staphylococcus warneri lipase 2

    NARCIS (Netherlands)

    van Kampen, M.D.; Rosenstein, R.; Götz, F.; Egmond, M.R.

    2010-01-01

    A gene encoding an extracellular lipase was identified in Staphylococcus warneri 863. The deduced lipase is organised as a prepro-protein and has significant similarity to other staphylococcal lipases. The mature part of the lipase was expressed with an N-terminal histidine tag in Escherichia coli,

  10. Role of Hsl7 in morphology and pathogenicity and its interaction with other signaling components in the plant pathogen Ustilago maydis.

    Science.gov (United States)

    Lovely, C Ben; Aulakh, Kavita Burman; Perlin, Michael H

    2011-07-01

    The phytopathogenic fungus Ustilago maydis undergoes a dimorphic transition in response to mating pheromone, host, and environmental cues. On a solid medium deficient in ammonium (SLAD [0.17% yeast nitrogen base without ammonium sulfate or amino acids, 2% dextrose, 50 μM ammonium sulfate]), U. maydis produces a filamentous colony morphology, while in liquid SLAD, the cells do not form filaments. The p21-activated protein kinases (PAKs) play a substantial role in regulating the dimorphic transition in fungi. The PAK-like Ste20 homologue Smu1 is required for a normal response to pheromone, via upregulation of pheromone expression, and virulence, and its disruption affects both processes. Our experiments suggest that Smu1 also regulates cell length and the filamentous response on solid SLAD medium. Yeast two-hybrid analysis suggested an Hsl7 homologue as a potential interacting partner of Smu1, and a unique open reading frame for such an arginine methyltransferase was detected in the U. maydis genome sequence. Hsl7 regulates cell length and the filamentous response to solid SLAD in a fashion opposite to that of Smu1, but neither overexpression nor disruption of hsl7 attenuates virulence. Simultaneous disruption of hsl7 and overexpression of smu1 lead to a hyperfilamentous response on solid SLAD. Moreover, only this double mutant strain forms filaments in liquid SLAD. The double mutant strain was also significantly reduced in virulence. A similar filamentous response in both solid and liquid SLAD was observed in strains lacking another PAK-like protein kinase involved in cytokinesis and polar growth, Cla4. Our data suggest that Hsl7 may regulate cell cycle progression, while both Smu1 and Cla4 appear to be involved in the filamentous response in U. maydis.

  11. LIPASES PRODUCED BY YEASTS: POWERFUL BIOCATALYSTS FOR INDUSTRIAL PURPOSES

    Directory of Open Access Journals (Sweden)

    Luiza Lux Lock

    2007-12-01

    Full Text Available The term “lipolytic enzymes” refers to the lipases and carboxylic ester hydrolases. Lipase production is widespread among yeasts, but few are capable of producing lipases with interesting characteristics and in sufficient amounts to be industrially useful. The literature concerning lipases produced by Candida rugosa, Yarrowia (Candida lipolytica, Candida antarctica and other emerging lipase-producing yeasts is reviewed. The use of recombinant lipases is discussed, with emphasis on the utilization of heterologous expression systems and design of chimeras. Finally, the three approaches that aim the improvement of lipase production or the modification of the substrate selectivity of the enzyme (medium engineering, biocatalyst engineering, and protein engineering are discussed.

  12. TLC bioautographic method for detecting lipase inhibitors.

    Science.gov (United States)

    Hassan, Abdel Moniem Sadek

    2012-01-01

    Bioautographic assays using TLC play an important role in the search for active compounds from plants. A TLC bioautographic assay has previously been established for the detection of acetylcholinesterase inhibitors but not for lipases. Development of a TLC bioautographic method for detecting lipase inhibitors in plant extracts. After migration of the plant extracts, the TLC plate was sprayed with α-naphtyl acetate and enzyme solutions before incubation at 37°C for 20 min. Finally, the solution of Fast Blue B salt was sprayed onto the TLC plate giving a purple background colouration. Lipase inhibitors were visualised as white spots on the TLC plates. Orlistat (a known lipase inhibitor) inhibited lipase down to 0.01 µg. Methanolic extracts of Camellia sinensis (L.) kuntz and Rosmarinus officinalis L after migration on TLC gave enzymatic inhibition when applied in amounts of 82 and 56 µg, respectively. On the other hand the methanolic extract of Morus alba leaves did not exhibit any lipase inhibitory activity. The screening test was able to detect lipase inhibition by pure reference substances and by compounds present in complex matrices, such as plant extracts. Copyright © 2011 John Wiley & Sons, Ltd.

  13. Advances in lipase-catalyzed esterification reactions.

    Science.gov (United States)

    Stergiou, Panagiota-Yiolanda; Foukis, Athanasios; Filippou, Michalis; Koukouritaki, Maria; Parapouli, Maria; Theodorou, Leonidas G; Hatziloukas, Efstathios; Afendra, Amalia; Pandey, Ashok; Papamichael, Emmanuel M

    2013-12-01

    Lipase-catalyzed esterification reactions are among the most significant chemical and biochemical processes of industrial relevance. Lipases catalyze hydrolysis as well as esterification reactions. Enzyme-catalyzed esterification has acquired increasing attention in many applications, due to the significance of the derived products. More specifically, the lipase-catalyzed esterification reactions attracted research interest during the past decade, due to an increased use of organic esters in biotechnology and the chemical industry. Lipases, as hydrolyzing agents are active in environments, which contain a minimum of two distinct phases, where all reactants are partitioned between these phases, although their distribution is not fixed and changes as the reaction proceeds. The kinetics of the lipase-catalyzed reactions is governed by a number of factors. This article presents a thorough and descriptive evaluation of the applied trends and perspectives concerning the enzymatic esterification, mainly for biofuel production; an emphasis is given on essential factors, which affect the lipase-catalyzed esterification reaction. Moreover, the art of using bacterial and/or fungal strains for whole cell biocatalysis purposes, as well as carrying out catalysis by various forms of purified lipases from bacterial and fungal sources is also reviewed.

  14. MICROBIAL LIPASES: PRODUCTION OF EXTRACELLULAR LIPASE ENZYME BY ALCALIGENES VISCOSUS (DOGE-1 STRAIN

    Directory of Open Access Journals (Sweden)

    P.Sekhar

    2012-05-01

    Full Text Available Industrially important extracellular lipase enzyme production was explored by utilizingmicrobial strain isolated from dairy effluents. Alcaligenes viscosus DOGE-1 strain isolated from dairywaste waters proved to produce extracellular lipase. Various growth factors were attempted to maximizethe lipase production by this strain. Growth factors like NH4PO4, Peptone, Urea coupled with peptone,KH2PO4, Olive oil and pH were found to be favored the maximum lipase production. This microbialstrain was found to have a high lipolytic activity.

  15. Lipase-catalyzed process for biodiesel production: protein engineering and lipase production.

    Science.gov (United States)

    Hwang, Hyun Tae; Qi, Feng; Yuan, Chongli; Zhao, Xuebing; Ramkrishna, Doraiswami; Liu, Dehua; Varma, Arvind

    2014-04-01

    Biodiesel is an environment-friendly and renewable fuel produced by transesterification of various feedstocks. Although the lipase-catalyzed biodiesel production has many advantages over the conventional alkali catalyzed process, its industrial applications have been limited by high-cost and low-stability of lipase enzymes. This review provides a general overview of the recent advances in lipase engineering, including both protein modification and production. Recent advances in biotechnology such as in protein engineering, recombinant methods and metabolic engineering have been employed but are yet to impact lipase engineering for cost-effective production of biodiesel. A summary of the current challenges and perspectives for potential solutions are also provided.

  16. Reduced plasma FFA availability increases net triacylglycerol degradation, but not GPAT or HSL activity, in human skeletal muscle.

    Science.gov (United States)

    Watt, Matthew J; Holmes, Anna G; Steinberg, Gregory R; Mesa, Jose L; Kemp, Bruce E; Febbraio, Mark A

    2004-07-01

    Intramuscular triacylglycerols (IMTG) are proposed to be an important metabolic substrate for contracting muscle, although this remains controversial. To test the hypothesis that reduced plasma free fatty acid (FFA) availability would increase IMTG degradation during exercise, seven active men cycled for 180 min at 60% peak pulmonary O(2) uptake either without (CON) or with (NA) prior ingestion of nicotinic acid to suppress adipose tissue lipolysis. Skeletal muscle and adipose tissue biopsy samples were obtained before and at 90 and 180 min of exercise. NA ingestion decreased (P HSL) activity (CON: 13.9 +/- 2.5, NA: 9.1 +/- 3.0 nmol.min(-1).mg protein(-1)). NA ingestion resulted in decreased whole body fat oxidation and increased carbohydrate oxidation. Despite the decreased whole body fat oxidation, net IMTG degradation was greater in NA compared with CON (net change: CON, 2.3 +/- 0.8; NA, 6.3 +/- 1.2 mmol/kg dry mass). The increased IMTG degradation did not appear to be due to reduced fatty acid esterification, because glycerol 3-phosphate activity was not different between trials and was unaffected by exercise (rest: 0.21 +/- 0.07; 180 min: 0.17 +/- 0.04 nmol.min(-1).mg protein(-1)). HSL activity was not increased from resting rates during exercise in either trial despite elevated plasma epinephrine, decreased plasma insulin, and increased ERK1/2 phosphorylation. AMP-activated protein kinase (AMPK)alpha1 activity was not affected by exercise or NA, whereas AMPKalpha2 activity was increased (P < 0.05) from rest during exercise in NA and was greater (P < 0.05) than in CON at 180 min. These data suggest that plasma FFA availability is an important mediator of net IMTG degradation, and in the absence of plasma FFA, IMTG degradation cannot maintain total fat oxidation. These changes in IMTG degradation appear to disassociate, however, from the activity of the key enzymes responsible for synthesis and degradation of this substrate.

  17. Photo-controlled deactivation of immobilised lipase

    NARCIS (Netherlands)

    Poloni, Claudia; Szymanski, Wiktor; Feringa, Ben L.

    2014-01-01

    Lipase from Candida rugosa was immobilised on a quartz surface using an azobenzene-containing, bifunctional linker, which allows deactivation of the immobilised enzyme by irradiation with visible light.

  18. Organic Solvent Tolerant Lipases and Applications

    National Research Council Canada - National Science Library

    Sharma, Shivika; Kanwar, Shamsher S

    2014-01-01

    ... the hydrolysis of triacylglycerol to glycerol and fatty acids [2]. Lipases find potential applications in bioprocesses largely due to their availability and stability in organic as well as in aqueo...

  19. Lipase and protease extraction from activated sludge

    DEFF Research Database (Denmark)

    Gessesse, Amare; Dueholm, Thomas; Petersen, Steffen B.

    2003-01-01

    of gentle and efficient enzyme extraction methods from environmental samples is very important. In this study we present a method for the extraction of lipases and proteases from activated sludge using the non-ionic detergent Triton X-100, EDTA, and cation exchange resin (CER), alone or in combination...... for the extraction of lipases and proteases from activated sludge. The sludge was continuously stirred in the presence of either buffer alone or in the presence of detergent and/or chelating agents. In all cases, a marked reduction in floc size was observed upon continuous stirring. However, no lipase activity...... and negligible protease activity was extracted in the presence of buffer alone, indicating that enzyme extraction was not due to shear force alone. The highest lipase activity was extracted using 0.1% Triton X-100 above which the activity was gradually decreasing. For proteases, the highest activity was obtained...

  20. Genetics Home Reference: familial lipoprotein lipase deficiency

    Science.gov (United States)

    ... Rare Disorders (NORD) RareConnect GeneReviews (1 link) Familial Lipoprotein Lipase Deficiency ClinicalTrials.gov (1 link) ClinicalTrials.gov Scientific Articles on PubMed (1 link) PubMed OMIM (1 link) ...

  1. Structure and Function of Lipase

    DEFF Research Database (Denmark)

    Skjold-Jørgensen, Jakob

    out to calculate the energydifference between the open and closed lid conformation for TlL and a selection of lid-variants (PaperIII). Here, a correlation between experimental and theoretical data was discovered supporting the notionlid plays a key role in governing activation at the interface...... towards an open conformation enabling the substrate to gain access, thus initiating catalysis.Lipases have been studied for decades and their functional features have drawn much attention withinindustrial applications since their first discovery. However, given that their molecular action takes placeat...... onthe activation mechanism. From characterization studies of these variants we have shown (Paper I) thatthe lid-region plays a crucial role in governing interfacial activation and enzymatic activity. Specifically,using a combination of spectroscopic and enzymatic activity-based methods we have...

  2. Structural characterization of MAPLE deposited lipase biofilm

    Energy Technology Data Exchange (ETDEWEB)

    Aronne, Antonio [Department of Chemical Engineering, Materials and Industrial Production, Università degli Studi di Napoli Federico II, Piazzale V. Tecchio 80, 80125 Napoli (Italy); Ausanio, Giovanni; Bloisi, Francesco [CNR-SPIN and Department of Physics, Università degli Studi di Napoli Federico II, Piazzale V. Tecchio 80, 80125 Napoli (Italy); Calabria, Raffaela [Istituto Motori-CNR, via G. Marconi 8, 80125 Napoli (Italy); Califano, Valeria, E-mail: v.califano@im.cnr.it [Istituto Motori-CNR, via G. Marconi 8, 80125 Napoli (Italy); Fanelli, Esther [Department of Chemical Engineering, Materials and Industrial Production, Università degli Studi di Napoli Federico II, Piazzale V. Tecchio 80, 80125 Napoli (Italy); Massoli, Patrizio [Istituto Motori-CNR, via G. Marconi 8, 80125 Napoli (Italy); Vicari, Luciano R.M. [CNR-SPIN and Department of Physics, Università degli Studi di Napoli Federico II, Piazzale V. Tecchio 80, 80125 Napoli (Italy)

    2014-11-30

    Highlights: • Lipase from Candida Rugosa was deposited by Matrix Assisted Pulsed Laser Evaporation (MAPLE) on KBr pellets, mica and glass substrate. • The deposited film was characterized morphologically and structurally by optical microscopy, SEM and FTIR analysis. • Results of characterization underlined a phenomenon of aggregation taking place. • The aggregation phenomenon was reversible since lipase showed activity in the transesterification reaction between soybean oil and isopropyl alcohol once detached from the substrate. - Abstract: Lipases (triacylglycerol ester hydrolases) are enzymes used in several industrial applications. Enzymes immobilization can be used to address key issues limiting widespread application at industrial level. Immobilization efficiency is related to the ability to preserve the native conformation of the enzyme. MAPLE (Matrix Assisted Pulsed Laser Evaporation) technique, a laser deposition procedure for treating organic/polymeric/biomaterials, was applied for the deposition of lipase enzyme in an ice matrix, using near infrared laser radiation. Microscopy analysis showed that the deposition occurred in micrometric and submicrometric clusters with a wide size distribution. AFM imaging showed that inter-cluster regions are uniformly covered with smaller aggregates of nanometric size. Fourier transform infrared spectroscopy was used for both recognizing the deposited material and analyzing its secondary structure. Results showed that the protein underwent reversible self-association during the deposition process. Actually, preliminary tests of MAPLE deposited lipase used for soybean oil transesterification with isopropyl alcohol followed by gas chromatography–mass spectrometry gave results consistent with undamaged deposition of lipase.

  3. Lipase-catalyzed reactions at different surfaces.

    Science.gov (United States)

    Reis, P; Holmberg, K; Debeche, T; Folmer, B; Fauconnot, L; Watzke, H

    2006-09-12

    Starting from gold chips, we have tailor-made three surfaces by the self-assembly monolayer technique: one entirely hydrophobic, one hydrophobic with dispersed carboxyl groups, and one hydrophilic, containing hydroxyl groups. Rhizomucor miehei lipase has been adsorbed to the hydrophobic and the hydrophilic surfaces and covalently bound to the surface containing carboxyl groups. The adsorption of two substrates-capric acid (decanoic acid) and monocaprin-on the lipase-covered surfaces was monitored by the surface plasmon resonance (SPR) technique. Biocatalysis was also performed in the SPR instrument by circulating a solution of the substrate, dissolved in an 85:15 water-glycerol mixture at a(w) = 0.81, through the instrument, thus exposing the capric acid or the monocaprin to the lipase-covered surfaces. The product composition was found to depend on the type of surface used. Lipase adsorbed at the hydrophilic surface favored hydrolysis, and capric acid was the main product formed when monocaprin was used as substrate. Lipase adsorbed at a hydrophobic surface and, in particular, lipase covalently bound to a hydrophobic surface favored condensation. More dicaprin than capric acid was formed in experiments with monocaprin as the substrate. Reactions performed outside the SPR instrument showed that small amounts of triglyceride were also formed under these conditions. We believe that this work constitutes the first example of the SPR instrument being used for in-situ biotransformation.

  4. Immobilised lipase for in vitro lipolysis experiments.

    Science.gov (United States)

    Phan, Stephanie; Salentinig, Stefan; Hawley, Adrian; Boyd, Ben J

    2015-04-01

    In vitro lipolysis experiments are used to assess digestion of lipid-based formulations, and probe solubilisation by colloidal phases during digestion. However, proteins and other biological components in the pancreatin often used as the lipase result in high-background scattering when interrogating structures using scattering approaches, complicating the resolution of colloidal structures. In this study, to circumvent this problem, a modified in vitro digestion model employing lipase immobilised on polymer beads, which allows for separation of the lipid digestion components during lipolysis, was investigated. Titration of the fatty acids released during digestion of medium chain triglycerides using pancreatin compared with immobilised lipase, combined with HPLC was used to follow the digestion, and small-angle X-ray scattering was used to determine colloidal structure formation. Digestion of medium chain triglycerides at the same nominal activity revealed that for the immobilised lipase, a longer digestion time was required to achieve the same extent of digestion. However, the same structural endpoint was observed, indicating that structure formation was not affected by the choice of lipase used. Lipolysis with immobilised lipase led to the reduction of parasitic scattering, resulting in clearer and more defined scattering from the structures generated by the lipolysis products. © 2015 Wiley Periodicals, Inc. and the American Pharmacists Association.

  5. Optimization of fermentation conditions for 1,3-propanediol production by marine Klebsiella pneumonia HSL4 using response surface methodology

    Science.gov (United States)

    Li, Lili; Zhou, Sheng; Ji, Huasong; Gao, Ren; Qin, Qiwei

    2014-09-01

    The industrially important organic compound 1,3-propanediol (1,3-PDO) is mainly used as a building block for the production of various polymers. In the present study, response surface methodology protocol was followed to determine and optimize fermentation conditions for the maximum production of 1,3-PDO using marine-derived Klebsiella pneumoniae HSL4. Four nutritional supplements together with three independent culture conditions were optimized as follows: 29.3 g/L glycerol, 8.0 g/L K2 HPO4, 7.6 g/L (NH4)2 SO4, 3.0 g/L KH2 PO4, pH 7.1, cultivation at 35°C for 12 h. Under the optimal conditions, a maximum 1,3-PDO concentration of 14.5 g/L, a productivity of 1.21 g/(L·h) and a conversion of glycerol of 0.49 g/g were obtained. In comparison with the control conditions, fermentation under the optimized conditions achieved an increase of 38.8% in 1,3-PDO concentration, 39.0% in productivity and 25.7% in glycerol conversion in flask. This enhancement trend was further confirmed when the fermentation was conducted in a 5-L fermentor. The optimized fermentation conditions could be an important basis for developing lowcost, large-scale methods for industrial production of 1,3-PDO in the future.

  6. Diacylglycerol synthesis by enzymatic glycerolysis: Screening of commercially available lipases

    DEFF Research Database (Denmark)

    Kristensen, Janni Brogaard; Xu, X.B.; Mu, Huiling

    2005-01-01

    yield (approx. 60 wt%) was achieved with Novozym 435 and Lipase PS-D after 7 h, and an equilibrium was obtained. Stepwise addition of glycerol allowed catalysis with Novozym CALB L (immobilized) to take place in spite of the hydrophilic carrier; however, the DAG yield was only 19 wt%. This result...... suggests that glycerol forms a layer around the hydrophilic lipase particles, limiting contact between the lipases and the hydrophobic oil phase. With glycerol absorbed on silica gel, all lipases catalyzed the glycerolysis reaction. Faster conversion of TAG was obtained with Lipase PS-D, Lipase AK...

  7. 肥胖症脂肪组织ATGL和HSL mRNA的表达与胰岛素敏感性的关系

    Institute of Scientific and Technical Information of China (English)

    江榕; 刘小莺; 郑培烝; 陈晓春; 刘礼斌

    2010-01-01

    目的:测定肥胖症患者内脏及皮下脂肪组织三酰甘油水解酶(ATGL)和激素敏感脂肪酶(HSL)mRNA的表达,探讨其与胰岛素敏感性的相关性。方法:应用荧光实时定量PCR法检测皮下、内脏脂肪组织ATGL和HSL mRNA的表达,应用HOMA-IR评估胰岛素敏感性,分析其与人体测量指标和代谢指标的相关性。结果:①肥胖组内脏与皮下脂肪组织ATGL mRNA及HSL mRNA表达量均低于正常对照组皮下脂肪组织;②皮下及内脏脂肪组织ATGL mRNA表达量与体质量指数(BMI)呈负相关;③皮下及内脏脂肪组织HSL mRNA表达量与BMI、腰围呈负相关。结论:①肥胖症脂肪组织ATGL mRNA及HSL mRNA表达降低;②ATGL及HSL与肥胖相关;③ATGL及HSL与胰岛素抵抗可能无明显相关性。

  8. Increased rate of whole body lipolysis before and after 9 days of bed rest in healthy young men born with low birth weight

    DEFF Research Database (Denmark)

    Alibegovic, Amra Ciric; Hojbjerre, Lise; Sonne, Mette

    2010-01-01

    Background: Individuals born with low birth weight (LBW) are at risk of developing type 2 diabetes mellitus (T2D), which may be precipitated by physical inactivity. Methods: 22 LBW and 23 controls were studied before and after bed rest by the hyperinsulinemic euglycemic clamp combined with indirect...... calorimetry, infusion of stable isotope tracers, preceded by an intravenous glucose tolerance test. Results: LBW subjects had similar BMI, but elevated abdominal obesity compared with controls. The basal rate of whole body lipolysis (WBL) was elevated in LBW subjects with and without correction for abdominal...... obesity before and after bed rest (all P = 0.01). Skeletal muscle hormone sensitive lipase (HSL) protein expression and phosphorylation at ser 565 were similar in the two groups. Bed rest resulted in a decrease in WBL and an increased skeletal muscle HSL Ser565 phosphorylation indicating a decreased HSL...

  9. Regulation of Lipolysis and Adipose Tissue Signaling during Acute Endotoxin-Induced Inflammation: A Human Randomized Crossover Trial

    DEFF Research Database (Denmark)

    Rittig, Nikolaj; Bach, Ermina; Thomsen, Henrik Holm

    2016-01-01

    lipolysis in adipose tissue and is associated with increased pHSL and signs of increased PLIN1 phosphorylation combined with a trend toward decreased insulin signaling. The combination of these mechanisms appear to be the driving forces behind the increased lipolysis observed in the early stages of acute...... = 0.001) compared with Placebo. These effects were associated with increased phosphorylation of hormone sensitive lipase (pHSL) at ser650 in adipose tissue (p = 0.03), a trend towards elevated pHSL at ser552 (p = 0.09) and cAMP-dependent protein kinase A (PKA) phosphorylation of perilipin 1 (PLIN1) (p......BACKGROUND: Lipolysis is accelerated during the acute phase of inflammation, a process being regulated by pro-inflammatory cytokines (e.g. TNF-α), stress-hormones, and insulin. The intracellular mechanisms remain elusive and we therefore measured pro- and anti-lipolytic signaling pathways...

  10. Differential phosphorylation of perilipin 1A at the initiation of lipolysis revealed by novel monoclonal antibodies and high content analysis.

    Science.gov (United States)

    McDonough, Patrick M; Maciejewski-Lenoir, Dominique; Hartig, Sean M; Hanna, Rita A; Whittaker, Ross; Heisel, Andrew; Nicoll, James B; Buehrer, Benjamin M; Christensen, Kurt; Mancini, Maureen G; Mancini, Michael A; Edwards, Dean P; Price, Jeffrey H

    2013-01-01

    Lipolysis in adipocytes is regulated by phosphorylation of lipid droplet-associated proteins, including perilipin 1A and hormone-sensitive lipase (HSL). Perilipin 1A is potentially phosphorylated by cAMP(adenosine 3',5'-cyclic monophosphate)-dependent protein kinase (PKA) on several sites, including conserved C-terminal residues, serine 497 (PKA-site 5) and serine 522 (PKA-site 6). To characterize perilipin 1A phosphorylation, novel monoclonal antibodies were developed, which selectively recognize perilipin 1A phosphorylation at PKA-site 5 and PKA-site 6. Utilizing these novel antibodies, as well as antibodies selectively recognizing HSL phosphorylation at serine 563 or serine 660, we used high content analysis to examine the phosphorylation of perilipin 1A and HSL in adipocytes exposed to lipolytic agents. We found that perilipin PKA-site 5 and HSL-serine 660 were phosphorylated to a similar extent in response to forskolin (FSK) and L-γ-melanocyte stimulating hormone (L-γ-MSH). In contrast, perilipin PKA-site 6 and HSL-serine 563 were phosphorylated more slowly and L-γ-MSH was a stronger agonist for these sites compared to FSK. When a panel of lipolytic agents was tested, including multiple concentrations of isoproterenol, FSK, and L-γ-MSH, the pattern of results was virtually identical for perilipin PKA-site 5 and HSL-serine 660, whereas a distinct pattern was observed for perilipin PKA-site 6 and HSL-serine 563. Notably, perilipin PKA-site 5 and HSL-serine 660 feature two arginine residues upstream from the phospho-acceptor site, which confers high affinity for PKA, whereas perilipin PKA-site 6 and HSL-serine 563 feature only a single arginine. Thus, we suggest perilipin 1A and HSL are differentially phosphorylated in a similar manner at the initiation of lipolysis and arginine residues near the target serines may influence this process.

  11. Differential phosphorylation of perilipin 1A at the initiation of lipolysis revealed by novel monoclonal antibodies and high content analysis.

    Directory of Open Access Journals (Sweden)

    Patrick M McDonough

    Full Text Available Lipolysis in adipocytes is regulated by phosphorylation of lipid droplet-associated proteins, including perilipin 1A and hormone-sensitive lipase (HSL. Perilipin 1A is potentially phosphorylated by cAMP(adenosine 3',5'-cyclic monophosphate-dependent protein kinase (PKA on several sites, including conserved C-terminal residues, serine 497 (PKA-site 5 and serine 522 (PKA-site 6. To characterize perilipin 1A phosphorylation, novel monoclonal antibodies were developed, which selectively recognize perilipin 1A phosphorylation at PKA-site 5 and PKA-site 6. Utilizing these novel antibodies, as well as antibodies selectively recognizing HSL phosphorylation at serine 563 or serine 660, we used high content analysis to examine the phosphorylation of perilipin 1A and HSL in adipocytes exposed to lipolytic agents. We found that perilipin PKA-site 5 and HSL-serine 660 were phosphorylated to a similar extent in response to forskolin (FSK and L-γ-melanocyte stimulating hormone (L-γ-MSH. In contrast, perilipin PKA-site 6 and HSL-serine 563 were phosphorylated more slowly and L-γ-MSH was a stronger agonist for these sites compared to FSK. When a panel of lipolytic agents was tested, including multiple concentrations of isoproterenol, FSK, and L-γ-MSH, the pattern of results was virtually identical for perilipin PKA-site 5 and HSL-serine 660, whereas a distinct pattern was observed for perilipin PKA-site 6 and HSL-serine 563. Notably, perilipin PKA-site 5 and HSL-serine 660 feature two arginine residues upstream from the phospho-acceptor site, which confers high affinity for PKA, whereas perilipin PKA-site 6 and HSL-serine 563 feature only a single arginine. Thus, we suggest perilipin 1A and HSL are differentially phosphorylated in a similar manner at the initiation of lipolysis and arginine residues near the target serines may influence this process.

  12. Differential Phosphorylation of Perilipin 1A at the Initiation of Lipolysis Revealed by Novel Monoclonal Antibodies and High Content Analysis

    Science.gov (United States)

    McDonough, Patrick M.; Hanna, Rita A.; Whittaker, Ross; Heisel, Andrew; Nicoll, James B.; Buehrer, Benjamin M.; Christensen, Kurt; Mancini, Maureen G.; Mancini, Michael A.; Edwards, Dean P.; Price, Jeffrey H.

    2013-01-01

    Lipolysis in adipocytes is regulated by phosphorylation of lipid droplet-associated proteins, including perilipin 1A and hormone-sensitive lipase (HSL). Perilipin 1A is potentially phosphorylated by cAMP(adenosine 3′,5′-cyclic monophosphate)-dependent protein kinase (PKA) on several sites, including conserved C-terminal residues, serine 497 (PKA-site 5) and serine 522 (PKA-site 6). To characterize perilipin 1A phosphorylation, novel monoclonal antibodies were developed, which selectively recognize perilipin 1A phosphorylation at PKA-site 5 and PKA-site 6. Utilizing these novel antibodies, as well as antibodies selectively recognizing HSL phosphorylation at serine 563 or serine 660, we used high content analysis to examine the phosphorylation of perilipin 1A and HSL in adipocytes exposed to lipolytic agents. We found that perilipin PKA-site 5 and HSL-serine 660 were phosphorylated to a similar extent in response to forskolin (FSK) and L-γ-melanocyte stimulating hormone (L-γ-MSH). In contrast, perilipin PKA-site 6 and HSL-serine 563 were phosphorylated more slowly and L-γ-MSH was a stronger agonist for these sites compared to FSK. When a panel of lipolytic agents was tested, including multiple concentrations of isoproterenol, FSK, and L-γ-MSH, the pattern of results was virtually identical for perilipin PKA-site 5 and HSL-serine 660, whereas a distinct pattern was observed for perilipin PKA-site 6 and HSL-serine 563. Notably, perilipin PKA-site 5 and HSL-serine 660 feature two arginine residues upstream from the phospho-acceptor site, which confers high affinity for PKA, whereas perilipin PKA-site 6 and HSL-serine 563 feature only a single arginine. Thus, we suggest perilipin 1A and HSL are differentially phosphorylated in a similar manner at the initiation of lipolysis and arginine residues near the target serines may influence this process. PMID:23405163

  13. Lipoprotein lipase gene variants: Association with acute myocardial ...

    African Journals Online (AJOL)

    Lipoprotein lipase gene variants: Association with acute myocardial infarction and lipid profiles. ... Therefore, genes involved in lipid and lipoprotein metabolism pathways such as lipoprotein lipase (LPL), are proper candidates ... Article Metrics.

  14. Immobilization of Isolated Lipase From Moldy Copra (Aspergillus Oryzae

    Directory of Open Access Journals (Sweden)

    Seniwati Dali

    2011-01-01

    Full Text Available Enzyme immobilization is a recovery technique that has been studied in several years, using support as a media to help enzyme dissolutions to the reaction substrate. Immobilization method used in this study was adsorption method, using specific lipase from Aspergillus oryzae. Lipase was partially purified from the culture supernatant of Aspergillus oryzae. Enzyme was immobilized by adsorbed on silica gel. Studies on free and immobilized lipase systems for determination of optimum pH, optimum temperature, thermal stability and reusability were carried out. The results showed that free lipase had optimum pH 8,2 and optimum temperature 35 °C while the immobilized lipase had optimum 8,2 and optimum temperature 45 °C. The thermal stability of the immobilized lipase, relative to that of the free lipase, was markedly increased. The immobilized lipase can be reused for at least six times.

  15. The modulation of pancreatic lipase activity by alginates

    OpenAIRE

    Wilcox, Matthew D.; Brownlee, Iain A.; Richardson, J. Craig; Dettmar, Peter W.; Jeffrey P. Pearson

    2014-01-01

    Alginates are comprised of mannuronic (M) and guluronic acid (G) and have been shown to inhibit enzyme activity. Pancreatic lipase is important in dietary triacylglycerol breakdown; reducing pancreatic lipase activity would reduce triacylglycerol breakdown resulting in lower amounts being absorbed by the body. Lipase activity in the presence of biopolymers was assessed by enzymatic assay using natural and synthetic substrates. Alginate inhibited pancreatic lipase by a maximum of 72.2% (±4.1) ...

  16. Mechanism of acetaldehyde-induced deactivation of microbial lipases

    OpenAIRE

    Jaeger Karl E; Eggert Thorsten; Franken Benjamin; Pohl Martina

    2011-01-01

    Abstract Background Microbial lipases represent the most important class of biocatalysts used for a wealth of applications in organic synthesis. An often applied reaction is the lipase-catalyzed transesterification of vinyl esters and alcohols resulting in the formation of acetaldehyde which is known to deactivate microbial lipases, presumably by structural changes caused by initial Schiff-base formation at solvent accessible lysine residues. Previous studies showed that several lipases were ...

  17. LIPOLYTIC ACTIVITY IN THE BLOOD AFTER LIPASE INGESTION,

    Science.gov (United States)

    The nature of the enzyme appearing in the blood after oral ingestion of pancreatic lipase has been studied by determining the effect of the presence...bile acid while the enzymic activity of both pancreatic lipase and the enzyme present in the blood after pancreatic lipase ingestion were greatly...enhanced by the presence of cholic acid. Although the rate of fat absorption has been shown to be affected by lipase ingestion , the possibility remains

  18. Lipases as biocatalysts for biodiesel production

    Directory of Open Access Journals (Sweden)

    Ognjanović Nevena D.

    2010-01-01

    Full Text Available Lipases can be used for a variety of biotechnological applications: synthesis of fine chemicals, therapeutics, agrochemicals, cosmetics, flavors, biopolymers and biodiesel. Biodiesel is an alternative fuel for diesel engines that is environmentally acceptable. Conventionally, biodiesel is produced by transesterification of triglycerides and short chain alcohols in the presence of an acid or an alkaline catalyst. There are several problems associated with this kind of production that can be resolved by using lipase as the biocatalyst. The usage of lipases has several advantages over the conventional chemical methods. It is considered as less energy intensive and environmentally friendly. However, there are two main obstacles associated with the effective utilization of lipases in the production of biodiesel. The main one is the cost of the enzyme and its poor stability in the presence of excess alcohol. Several strategies are proposed to overcome these drawbacks: immobilization of lipases, stepwise addition of alcohol, and the usage of novel acyl acceptors and the usage of whole cell biocatalysts.

  19. Stereoselectivity of lipases: esterification reactions of octadecylglycerol.

    Science.gov (United States)

    Meusel, D; Weber, N; Mukherjee, K D

    1992-04-01

    Stereoselectivity of several triacylglycerol lipases (EC 3.1.1.3) has been investigated in the enzymatic esterification of rac-1-O-octadecylglycerol with oleic acid in the presence of organic solvents, such as hexane. X-1(3)-O-Octadecylmonooleoylglycerols were the only products formed with most lipases; considerable proportions of X-1(3)-O-octadecyldioleoylglycerols were also formed with the lipase from Candida cylindracea. The mixtures of unesterified enantiomeric substrates, i.e., X-1(3)-O-octadecylglycerols were converted to their 3,5-dinitrophenylurethane derivatives and subsequently resolved into sn-1 and sn-3 enantiomers by HPLC on a chiral stationary phase (Sumichiral OA 2100). The data on enantiomeric excess (ee) and enantiomeric ratio (E) in the unesterified substrate revealed for the lipases from porcine pancreas, Rhizopus sp., Pseudomonas sp., Candida cylindracea, Chromobacterium viscosum and Penicillium cyclopium a distinct preference for 1-O-octadecyl-sn-glycerol over its enantiomer indicating stereoselectivity for the sn-3 position. For the lipase from Rhizomucor miehei a slight stereoselectivity for the sn-1 position was observed. Solvents, such as diethyl ether and dichloromethane, strongly inhibited the esterification reaction, but the enzymatic activity could be restored upon removal of such solvents by washing with hexane indicating reversible inhibition.

  20. Comparison of lipases for in vitro models of gastric digestion

    DEFF Research Database (Denmark)

    Sassene, P J; Fanø, M; Mu, H;

    2016-01-01

    The aim of this study was to find a lipase suitable as a surrogate for Human Gastric Lipase (HGL), since the development of predictive gastrointestinal lipolysis models are hampered by the lack of a lipase with similar digestive properties as HGL. Three potential surrogates for HGL; Rhizopus Oryzae...

  1. Wild-type and specific mutant androgen receptor mediates transcription via 17β-estradiol in sex hormone-sensitive cancer cells.

    Science.gov (United States)

    Susa, Takao; Ikaga, Reina; Kajitani, Takashi; Iizuka, Masayoshi; Okinaga, Hiroko; Tamamori-Adachi, Mimi; Okazaki, Tomoki

    2015-07-01

    We previously encountered regulatory processes wherein dihydrotestosterone (DHT) exerted its inhibitory effect on parathyroid hormone-related protein (PTHrP) gene repression through the estrogen receptor (ER)α, but not the androgen receptor (AR), in breast cancer MCF-7 cells. Here, we investigated whether such aberrant ligand-nuclear receptor (NR) interaction is present in prostate cancer LNCaP cells. First, we confirmed that LNCaP cells expressed large amounts of AR at negligible levels of ERα/β or progesterone receptor. Both suppression of PTHrP and activation of prostate-specific antigen genes were observed after independent administration of 17β-estradiol (E2), DHT, or R5020. Consistent with the notion that the LNCaP AR lost its ligand specificity due to a mutation (Thr-Ala877), experiments with siRNA targeting the respective NR revealed that the AR monopolized the role of the mediator of shared hormone-dependent regulation, which was invariably associated with nuclear translocation of this mutant AR. Microarray analysis of gene regulation by DHT, E2, or R5020 disclosed that more than half of the genes downstream of the AR (Thr-Ala877) overlapped in the LNCaP cells. Of particular interest, we realized that the AR (wild-type [wt]) and AR (Thr-Ala877) were equally responsible for the E2-AR interactions. Fluorescence microscopy experiments demonstrated that both EGFP-AR (wt) and EGFP-AR (Thr-Ala877) were exclusively localized within the nucleus after E2 or DHT treatment. Furthermore, reporter assays revealed that some other cancer cells exhibited aberrant E2-AR (wt) signaling similar to that in the LNCaP cells. We herein postulate the presence of entangled interactions between wt AR and E2 in certain hormone-sensitive cancer cells.

  2. B56alpha/protein phosphatase 2A inhibits adipose lipolysis in high-fat diet-induced obese mice.

    Science.gov (United States)

    Kinney, Brice P; Qiao, Liping; Levaugh, Justin M; Shao, Jianhua

    2010-08-01

    Lipolysis and lipogenesis are two opposite processes that control lipid storage in adipocytes. Impaired adipose lipolysis has been observed in both obese human subjects and animal models. This study investigated the mechanisms underlying impaired adipose lipolysis in a high-fat diet-induced obese (DIO) mouse model. DIO models were created using male C57BL/6 mice. Our results show that beta3 adrenergic receptor-specific agonist BRL37344 induced adipose lipolysis was significantly blunted in DIO mice. The levels of Ser660 phosphorylation of hormone-sensitive lipase (HSL) were significantly decreased in the epididymal fat of DIO mice. However, protein levels of HSL, adipose triglyceride lipase and its coactivator comparative gene identification-58 were similar between DIO and control mice. It is known that upon lipolytic hormone stimulation, protein kinase A phosphorylates HSL Ser660 and activates HSL, whereas protein phosphatase 2A (PP2A) dephosphorylates and inactivates HSL. Interestingly, our study shows that high-fat feeding did not alter epididymal fat cAMP and protein kinase A protein levels but significantly increased the expression of the alpha-isoform of PP2A regulatory subunit B' (B56alpha). To study the role of B56alpha in obesity-associated lipolytic defect, B56alpha was overexpressed or knocked down by adenovirus-mediated gene transduction in cultured 3T3-L1CARDelta1 adipocytes. Overexpression of B56alpha significantly decreased HSL Ser660 phosphorylation. In contrast, knocking down B56alpha increased hormone-stimulated HSL activation and lipolysis in mature 3T3-L1CARDelta1 adipocytes. These results strongly suggest that elevated B56alpha/PP2A inhibits HSL and lipolysis in white adipose tissue of DIO mice.

  3. Lipoprotein lipase and endothelial lipase in human testis and in germ cell neoplasms

    DEFF Research Database (Denmark)

    Nielsen, J E; Lindegaard, M L; Friis-Hansen, L;

    2009-01-01

    The aim of this study was to investigate endothelial lipase (EL, LIPG) and lipoprotein lipase (LPL) mRNA and protein expression in normal human testis and testicular germ cell tumours (GCT). Both EL and LPL were expressed in normal seminiferous tubules and in the interstitial compartment. EL m....... The results suggest that both EL and LPL participate in the supply of nutrients and steroidogenesis in the testes, and that especially EL may be important for the supply of cholesterol for testosterone production in the Leydig cells. The partial cellular separation of the expression of the two lipases...

  4. Lipase biocatalysis for useful biodegradable products

    Energy Technology Data Exchange (ETDEWEB)

    Linko, Y.Y.; Wang, Zhuo Lin; Uosukainen, E.; Seppaelae, J. [Helsinki Univ. of Technology, Espoo (Finland); Laemsae, M. [Raisio Group Oil Milling Industry, Raisio (Finland)

    1996-12-31

    It was shown that lipases can be used as biocatalysts in the production of useful biodegradable compounds such as 1-butyl oleate by direct esterification of butanol and oleic acid to decrease viscosity of biodiesel in winter use. By enzymic transesterification, a mixture of 2-ethyl-1-hexyl esters from rapeseed oil fatty acids can be obtained in good yields for use as a solvent, and of trimethylolpropane esters for use as a lubricant. Finally, it was demonstrated that polyesters with a mass average molar mass in excess of 75,000 g mol{sup -}1 can be obtained by esterification or transesterification by using lipase as biocatalyst. (author) (3 refs.)

  5. Porcine Pancreatic Lipase Related Protein 2 has High Triglyceride Lipase Activity in the Absence of Colipase

    OpenAIRE

    2013-01-01

    Efficient dietary fat digestion is essential for newborns who consume more dietary fat per body weight than at any other time of life. In many mammalian newborns, pancreatic lipase related protein 2 (PLRP2) is the predominant duodenal lipase. Pigs may be an exception since PLRP2 expression has been documented in the intestine but not in the pancreas. Because of the differences in tissue-specific expression, we hypothesized that the kinetic properties of porcine PLRP2 would differ from those o...

  6. Diet quality determines lipase gene expression and lipase/esterase activity in Daphnia pulex

    Directory of Open Access Journals (Sweden)

    Apostolos-Manuel Koussoroplis

    2017-02-01

    Full Text Available We studied the short- (12 h and long-term (144 h response of Daphnia pulex lipases to quality shifts in diets consisting of different mixtures of the green alga Scenedesmus with the cyanobacterium Synechococcus, two species with contrasting lipid compositions. The lipase/esterase activity in both the gut and the body tissues had fast responses to the diet shift and increased with higher dietary contributions of Synechococcus. When screening the Daphnia genome for TAG lipases, we discovered a large gene-family expansion of these enzymes. We used a subset of eight genes for mRNA expression analyses and distinguished between influences of time and diet on the observed gene expression patterns. We identified five diet-responsive lipases of which three showed a sophisticated short- and long-term pattern of expression in response to small changes in food-quality. Furthermore, the gene expression of one of the lipases was strongly correlated to lipase/esterase activity in the gut suggesting its potentially major role in digestion. These findings demonstrate that the lipid-related enzymatic machinery of D. pulex is finely tuned to diet and might constitute an important mechanism of physiological adaptation in nutritionally complex environments.

  7. Neurohypophyseal hormone-responsive renal adenylate cyclase. IV. A random-hit matrix model for coupline in a hormone-sensitive adenylate cyclase system.

    Science.gov (United States)

    Bergman, R N; Hechter, O

    1978-05-10

    (A), uniformly observed in different membrane preparations: 2) variable hormone concentration-dependent trajectories of approach to the final steady state A:O value (A:Oss) which may be either monophasic or biphasic; 3) the loss of intrinsic adenylate cyclase activity observed in bovine membranes for a series of NHH analogs with progressively diminishing affinity for receptors. The model represents an explicit theory of coupling where a successive series of temporal events are quantitatively related to each other and privide major constraints to any interpretation of the molecular organization of receptors and adenylate cyclase units in membranes. The model excludes a number of mechanistic proposals and suggests a new hypothesis for membrane coupling with features which may be generally applicable to other hormone-sensitive adenylate cyclase systems.

  8. Amylase and lipase values in normal subjects

    NARCIS (Netherlands)

    Riet, H.G. van

    In 146 hospitalized individuals (71 men and 75 women), the simultaneous fasting serum amylase and lipase concentrations and the urinary amylase excretion per 24 h were determined. Patients with pancreatic abnormalities or other diseases which might produce abnormal enzyme values were ruled out from

  9. Microbial lipases: Production, properties and biotechnological applications

    Directory of Open Access Journals (Sweden)

    Josana Maria Messias

    2011-09-01

    Full Text Available Lipases belong to the group of hydrolases that catalyze the hydrolysis of triacylglycerol lipids to free fatty acids and glycerol. They have significant potential biotechnological applications in catalyzing organic synthesis reactions in non-aqueous solvents using simplified procedures resulting in conversions of high yields. Lipase production has conventionally been performed by submerged fermentation; however, solid-state fermentation processes have been prominent when residues are used as substrates because they serve as low-cost nutrient sources. Microbial lipases can be used as additives in foods to modify and enhance organoleptic properties, as well as in detergents to hydrolyse fats in the treatment of oily effluents, and also have value for pharmaceutical, cosmetic, agrochemical, and oil chemical industries. More recently, they are used in transesterification reactions to convert plant seed oils into biodiesel. The objective of this work was to review the published literature on the production, properties and applications of microbial lipases, and its biotechnological role in producing biodiesel.

  10. New extremophilic lipases and esterases from metagenomics.

    Science.gov (United States)

    López-López, Olalla; Cerdán, Maria E; González Siso, Maria I

    2014-01-01

    Lipolytic enzymes catalyze the hydrolysis of ester bonds in the presence of water. In media with low water content or in organic solvents, they can catalyze synthetic reactions such as esterification and transesterification. Lipases and esterases, in particular those from extremophilic origin, are robust enzymes, functional under the harsh conditions of industrial processes owing to their inherent thermostability and resistance towards organic solvents, which combined with their high chemo-, regio- and enantioselectivity make them very attractive biocatalysts for a variety of industrial applications. Likewise, enzymes from extremophile sources can provide additional features such as activity at extreme temperatures, extreme pH values or high salinity levels, which could be interesting for certain purposes. New lipases and esterases have traditionally been discovered by the isolation of microbial strains producing lipolytic activity. The Genome Projects Era allowed genome mining, exploiting homology with known lipases and esterases, to be used in the search for new enzymes. The Metagenomic Era meant a step forward in this field with the study of the metagenome, the pool of genomes in an environmental microbial community. Current molecular biology techniques make it possible to construct total environmental DNA libraries, including the genomes of unculturable organisms, opening a new window to a vast field of unknown enzymes with new and unique properties. Here, we review the latest advances and findings from research into new extremophilic lipases and esterases, using metagenomic approaches, and their potential industrial and biotechnological applications.

  11. Gastric Lipase Secretion in Children with Gastritis

    Directory of Open Access Journals (Sweden)

    Krystyna Sztefko

    2013-07-01

    Full Text Available Gastric lipase is one of the prepancreatic lipases found in some mammalian species and in humans. Our knowledge of the hormonal regulation of gastric lipase secretion in children and adolescents is still very limited. The aim of this study was to compare the activity of human gastric lipase (HGL in gastric juice in healthy adolescents and in patients with gastritis. The adolescents were allocated to three groups: the first including patients with Helicobacter pylori gastritis (HPG; n = 10, the second including patients with superficial gastritis caused by pathogens other than H. pylori (non-HPG; n = 14 and the control group including healthy adolescents (n = 14. Activity of HGL was measured in gastric juice collected during endoscopy. Plasma concentrations of cholecystokinin (CCK, glucagon-like peptide-1 (GLP-1 and glucose-dependent insulinotropic peptide (GIP were measured in all adolescents. Activity of HGL in the non-HPG group was significantly lower than in the HPG group (p < 0.005 and the control group (p < 0.005. Mean plasma GIP levels in the control group were lower than in the non-HPG group (p < 0.003 and the HPG group (p < 0.01. We conclude that the regulation of HGL secretion by GLP-1 and CCK is altered in patients with gastritis. Moreover, GIP is a potent controller of HGL activity, both in healthy subjects and in patients with gastritis.

  12. Gastric lipase secretion in children with gastritis.

    Science.gov (United States)

    Tomasik, Przemyslaw J; Wędrychowicz, Andrzej; Rogatko, Iwona; Zając, Andrzej; Fyderek, Krzysztof; Sztefko, Krystyna

    2013-07-29

    Gastric lipase is one of the prepancreatic lipases found in some mammalian species and in humans. Our knowledge of the hormonal regulation of gastric lipase secretion in children and adolescents is still very limited. The aim of this study was to compare the activity of human gastric lipase (HGL) in gastric juice in healthy adolescents and in patients with gastritis. The adolescents were allocated to three groups: the first including patients with Helicobacter pylori gastritis (HPG; n = 10), the second including patients with superficial gastritis caused by pathogens other than H. pylori (non-HPG; n = 14) and the control group including healthy adolescents (n = 14). Activity of HGL was measured in gastric juice collected during endoscopy. Plasma concentrations of cholecystokinin (CCK), glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic peptide (GIP) were measured in all adolescents. Activity of HGL in the non-HPG group was significantly lower than in the HPG group (p gastritis. Moreover, GIP is a potent controller of HGL activity, both in healthy subjects and in patients with gastritis.

  13. In silico modeling of lipase H

    African Journals Online (AJOL)

    Amara

    2013-04-17

    Apr 17, 2013 ... ... and tertiary structure lipase H gene (LIPH) at molecular level were analyzed in the current study. ... Tertiary structure of LIPH was predicted through homology modeling. Mutations ... absence of axillary hair, and affected males usually have .... color, while in mutated LIPH, amino acid is shown in pink color.

  14. Amylase and lipase values in normal subjects

    NARCIS (Netherlands)

    Riet, H.G. van

    1968-01-01

    In 146 hospitalized individuals (71 men and 75 women), the simultaneous fasting serum amylase and lipase concentrations and the urinary amylase excretion per 24 h were determined. Patients with pancreatic abnormalities or other diseases which might produce abnormal enzyme values were ruled out from

  15. Acrylamide-quenching of Rhizomucor miehei lipase.

    Science.gov (United States)

    Stobiecka, Agnieszka

    2005-07-01

    Steady-state and time-resolved fluorescence-quenching measurements have been performed to study multitryptophan lipase from filamentous fungus Rhizomucor miehei. Using the steady-state acrylamide fluorescence quenching data and the fluorescence-quenching-resolved-spectra (FQRS) method, the total emission spectrum of native ("closed-lid") lipase has been decomposed into two distinct spectral components accessible to acrylamide. According to FQRS analysis, more quenchable component has a maximum of fluorescence emission at about 352 nm whereas less quenchable component emits at about 332 nm. The redder component participates in about 60-64% of the total lipase fluorescence and may be characterized by the dynamic and static quenching constants equal to K(1) = 3.75 M(-1) and V(1) = 1.12 M(-1), respectively. The bluer component is quenchable via dynamic mechanism with K(2) = 1.97 M(-1). Significant difference in the values of acrylamide bimolecular rate quenching constants estimated for redder and bluer component (i.e., k(q) = 1.2 x 10 (9) M(-1)s (-1) vs. k(q) = 4.3 x 10(8) M(-1) s(-1), respectively), suggests that tryptophan residues in fungal lipase are not uniformly exposed to the solvent.

  16. Lipoprotein lipase isoelectric point isoforms in humans

    DEFF Research Database (Denmark)

    Badia-Villanueva, M.; Carulla, P.; Carrascal, M.

    2014-01-01

    Lipoprotein lipase (LPL) hydrolyzes circulating triacylglycerols (TAG) into free fatty acids and glycerol. It is present in almost all tissues and its tissue-specific regulation directs the flow of circulating TAG in the body. We demonstrated in a previous study that, in rat heart and post-hepari...

  17. 日粮类型对猪肌肉组织HSL mRNA和Obese mRNA基因表达的影响%Effects of Diet Type on Expression of HSL mRNA and Obese mRNA in Muscle Tissue of Pigs

    Institute of Scientific and Technical Information of China (English)

    王安娜; 邓蓉; 张定红; 张洁

    2012-01-01

    Twenty Duroc x Landrace x large three crossbred pigs were allocated randomLy into two groups and fed soybean diet and miscellane- ous diet ( double low rapeseed meal and soybean meal). The pigs weight were reached about one hundred kilogram when slaughter test slaughtered and determined lean lean rate, in addition, taked the longissimus dorsi muscle tissue, The levels of HSL mRNA and Obese mRNA were detected by RT - PCR to explore relationship of diet type and gene expression. The results showed that miscellaneous meal with soybean meal in the diets significantly in- creased HSL expression of mRNA, decreased Obese expression of mRNA ( miscellaneous meal with soybean meal in the diets significantly increased HSL expression of mRNA, decreased Obese expression of mRNA ( P 〈 0.05 ) , Between soybean diet and miscellaneous diet on pig lean meat rate has no significant impact, but the actual data showed that pig fed miscellaneous diet showed a tendency to increase 3.5% ( P 〉 0.05 ) , comparisoning of pig fed soybean diet.%选用20 kg左右的"杜×长×大"外三元杂交仔猪20头,随机分为2组,分别饲喂豆粕型和杂粕(双低菜籽粕+豆粕)型2种不同类型的日粮,饲养猪至体重达到100 kg左右时屠宰测定瘦肉率,取背最长肌肌肉组织,采用RT-PCR技术检测分析HSL mRNA和Obese mRNA表达情况,以探索出日粮类型与基因表达的关系。结果显示,杂粕日粮较豆粕日粮可显著增加HSL mRNA表达量、减少Obese mRNA表达量(P〈0.05),2种日粮类型虽对猪瘦肉率无显著性影响,但是从实际值来看,杂粕日粮表现出提高趋势,较豆粕日粮提高3.5%(P〉0.05)。

  18. Infrared study of the bacterial autoinducer N-hexanoyl-homoserine lactone (C6-HSL) in the gas-phase, water, and octanol solutions.

    Science.gov (United States)

    Kim, Jiah; Kim, Seong Keun; Grégoire, Gilles; Manil, Bruno; Schermann, Jean Pierre

    2011-08-25

    The N-hexanoyl-homoserine lactone (C6-HSL) molecule has been investigated by means of infrared multiphoton dissociation (IRMPD) and Fourier-transform infrared spectroscopy (FT-IR) under different conditions in an attempt to mimic biological situations encountered in communication between bacteria for quorum sensing. The protonated molecular ion was studied in the gas-phase that corresponds to a solvent-free situation somewhat analogous to that encountered in the receptor. The simulation of the IRMPD spectrum of the isolated ion was then conducted by means of quantum chemistry calculations in vacuum. In the case of the neutral species, the FT-IR spectra were recorded in D(2)O, mimicking the cytosolic and extracellular media as well as in 1-octanol that is often used for simulation of cell membranes. The interpretation was conducted by considering a C6-HSL molecule in its endo or exo conformation hydrogen-bonded to, respectively, six D(2)O and four 1-octanol molecules. A satisfying agreement with the experimental FT-IR studies conducted in solution at room temperature was obtained as long as a continuum IEFPCM model was added to the explicit solvent environment.

  19. In silico and experimental characterization of chimeric Bacillus thermocatenulatus lipase with the complete conserved pentapeptide of Candida rugosa lipase.

    Science.gov (United States)

    Hosseini, Mostafa; Karkhane, Ali Asghar; Yakhchali, Bagher; Shamsara, Mehdi; Aminzadeh, Saeed; Morshedi, Dena; Haghbeen, Kamahldin; Torktaz, Ibrahim; Karimi, Esmat; Safari, Zahra

    2013-02-01

    Lipases are one of the highest value commercial enzymes as they have broad applications in detergent, food, pharmaceutical, and dairy industries. To provide chimeric Bacillus thermocatenulatus lipase (BTL2), the completely conserved pentapeptide (¹¹²Ala-His-Ser-Gln-Gly¹¹⁶) was replaced with similar sequences (²⁰⁷Gly-Glu-Ser-Ala-Gly²¹¹) of Candida rugosa lipase (CLR) at the nucleophilic elbow region. For this purpose, three mutations including A112G, H113E, and Q115A were inserted in the conserved pentapeptide sequence of btl2 gene. Based on the crystal structures of 2W22, the best structure of opened form of the chimeric lipases were garnered using the MODELLER v9.10 software. The native and chimeric lipases were docked to a set of ligands, and a trial version of Molegro Virtual Docker (MVD) software was used to obtain the energy values. Docking results confirmed chimeric lipase to be better than the native lipase. Following the in silico study, cloning experiments were conducted and expression of native and chimeric btl2 gene in Pichia pastoris was performed. The native and chimeric lipases were purified, and the effect of these mutations on characteristics of chimeric lipase studied and then compared with those of native lipase. Chimeric lipase exhibited 1.6-fold higher activity than the native lipase at 55 °C. The highest percentage of both lipases activity was observed at 60 °C and pH of 8.0. The ion Ca²⁺ slightly inhibited the activity of both lipases, whereas the organic solvent enhanced the lipase stability of chimeric lipase as compared with the native lipase. According to the results, the presence of two glycine residues at the conserved pentapeptide region of this chimeric lipase (¹¹²Gly-Glu-Ser-Ala-Gly¹¹⁶) may increase the flexibility of the nucleophilic elbow region and affect the enzyme activity level.

  20. FRAKSINASI ENZIM LIPASE DARI ENDOSPERM KELAPA DENGAN METODE SALTING OUT (Lipase fractionation of Coconut Endosperm by Salting out Method

    Directory of Open Access Journals (Sweden)

    Moh. Su'i

    2014-02-01

    Full Text Available This research learns about fractionation of lipases activity from coconut endosperm by using ammonium sulphate of 0–15%; 15-30 %, 30–45 %, 45–60 %, 60–75 % and 75–90 %. The results showed that the fractions of 0–15% ; 30–45 %, 45–60 % and 60–75 % have lipase activity. Meanwhile, the highest activity was fractions of 60-75%. fractions of 15-30% and 75-90%  have no lipase enzym activity. Molecule weigh of lipase enzyme was 72 kDa. Keywords: Lipases, endosperm, coconut, fractionation, ammonium sulphate   ABSTRAK Penelitian ini mempelajari fraksinasi enzim lipase dari endosperm kelapa menggunakan ammonium sulfat. fraksinasi dilakukan dengan variasi konsentrasi ammonium sulfat 0–15% ; 15-30%; 30–45 %, 45–60 %, 60–75 % dan 75–90 %. Hasil penelitian menunjukkan bahwa enzim lipase terdapat pada fraksi 0–15% ; 30–45 %, 45–60 % dan fraksi 60–75 % dengan aktivitas enzim tertinggi pada fraksi 60-75%. Sedangkan fraksi 15-30% dan 75-90% tidak ada enzim lipase. Berat molekul enzim lipase pada semua fraksi 72 kDa. Kata kunci: Lipase, endosperm, fraksinasi, ammonium sulfat

  1. Comparative studies of canine colipase and lipases from bovine, porcine, canine, human and rat pancreases.

    Science.gov (United States)

    Lee, P C

    1978-01-01

    1. Colipase was purified from canine pancreatic juice and found to have certain specificity in its reaction with various pancreatic lipases. 2. This colipase will stimulate the lipolytic activities of lipases isolated from canine, bovine and porcine pancreas but not lipases from a fungus, or from human and rat pancreases. 3. Characterization of these lipases showed (a) the molecular dimension of rat lipase is very different from the other lipases; (b) the pIs of canine, porcine and bovine lipases are almost identical but different from the pIs of rat, human and Candida (a fungus) lipases; and (c) the antiserum prepared against canine lipase will also react with lipases from human, hog and cow pancreases but not with rat and Candida lipases. 4. These physical differences can explain partly the difference in reaction between the various lipases and the canine colipase.

  2. Dependency of water concentration on ethanolysis of trioleoylglycerol by lipases

    DEFF Research Database (Denmark)

    Piyatheerawong, W.; Iwasaki, Y; Xu, Xuebing

    2004-01-01

    The effects of water concentration on ethanolysis of trioleoylglycerol catalyzed by four different lipases were studied. The target product of the ethanolysis was 2-monooleoylglycerol (2-MO). Novozym 435 (a commercially available preparation of immobilized Candida antarctica lipase B, CALB......) exhibited both the highest product yield and the reaction rate at very low (less than 1 wt.%) free water concentration. Its catalytic activity did not drop even in dry state, i.e. in the system of dry CALB in dry ethanol (water concentration was ca. 0.1 wt.%). In contrast, other three immobilized lipases...... tested (Rhizomucor miehei lipase, Burkholderia cepacia lipase and Thermomyces lanuginosus lipase) required larger amounts of free water (ca. 7-9 wt.%) for their best performance and exhibited no ethanolysis reaction at low free water concentrations. The CALB's anomalous behavior was also observed...

  3. Porcine pancreatic lipase related protein 2 has high triglyceride lipase activity in the absence of colipase.

    Science.gov (United States)

    Xiao, Xunjun; Ross, Leah E; Sevilla, Wednesday A; Wang, Yan; Lowe, Mark E

    2013-09-01

    Efficient dietary fat digestion is essential for newborns who consume more dietary fat per body weight than at any other time of life. In many mammalian newborns, pancreatic lipase related protein 2 (PLRP2) is the predominant duodenal lipase. Pigs may be an exception since PLRP2 expression has been documented in the intestine but not in the pancreas. Because of the differences in tissue-specific expression, we hypothesized that the kinetic properties of porcine PLRP2 would differ from those of other mammals. To characterize its properties, recombinant porcine PLRP2 was expressed in HEK293T cells and purified to homogeneity. Porcine PLRP2 had activity against tributyrin, trioctanoin and triolein. The activity was not inhibited by bile salts and colipase, which is required for the activity of pancreatic triglyceride lipase (PTL), minimally stimulated PLRP2 activity. Similar to PLRP2 from other species, PLRP2 from pigs had activity against galactolipids and phospholipids. Importantly, porcine PLRP2 hydrolyzed a variety of dietary substrates including pasteurized human mother's milk and infant formula and its activity was comparable to that of PTL. In conclusion, porcine PLRP2 has broad substrate specificity and has high triglyceride lipase activity even in the absence of colipase. The data suggest that porcine PLRP2 would be a suitable lipase for inclusion in recombinant preparations for pancreatic enzyme replacement therapy.

  4. Expression and Mutagenesis studies of Candida antactica lipase B

    OpenAIRE

    Rotticci-Mulder, Johanna C.

    2003-01-01

    Recombinant Candida antarctica lipase B was successfullyproduced in the methylotropic yeast Pichia pastoris. Thespecific activities of Candida antarctica lipase B produced inPichia pastoris and commercial Candida antarctica lipase B fromNovozymes were the same. In shake-flask cultivations theexpression levels were about 25 mg L-1. Production levels couldbe increased to 1.5 g L-1, using a fermentor. A model tosimulate growth and oxygen consumption was described. The highcell density growth cou...

  5. Pressure stability of lipases and their use in different systems

    OpenAIRE

    Knez, Željko; Leitgeb, Maja

    2015-01-01

    For the investigation of the solvent impact on the enzymes, lipases from different sources (Pseudomonas fluorescences, Rhizopus javanicus, Rhizopus niveus, Candida rugose and Porcine pancreas) were used. Stability and activity of these lipases in aqueous medium in supercritical $CO_2$ and liquid propane at 100 bar and 40°C were studied. On the basis of previous results lipases were used for their application in two different systems. The application of the polysulphone membrane in the cont...

  6. Lipase-catalyzed production of lysophospholipids

    Directory of Open Access Journals (Sweden)

    Mnasri Taha

    2017-07-01

    Full Text Available Lysophospholipids, such as lysophosphatidic acid or lysophosphatidylcholine, are important bioactive lipids, involved in various normal and pathological cellular processes. They also have industrial and pharmaceutical uses such as emulsifiers or components of drug delivery systems. Lipases, which natural substrates are long chain triacylglycerols, are important biocatalysts for organic synthesis mainly due to their broad substrate specificity and their ability to display high catalytic activity in organic media. This paper describes the various lipase-catalyzed reactions implemented for the production of lysophospholipids. They include hydrolysis or alcoholysis of phospholipids and acylation of the glycerophosphoryl moiety. Special emphasis is made on our work dealing with the production of lysophospholipids rich in dososahexaenoic acid, an important dietary polyunsaturated fatty acid via the hydrolysis of phospholipids extracted from the microalga Isochrysis galbana.

  7. Lipase and protease extraction from activated sludge

    DEFF Research Database (Denmark)

    Gessesse, Amare; Dueholm, Thomas; Petersen, Steffen B.

    2003-01-01

    In the process of wastewater treatment hydrolysis of polymeric substances is the first and rate-limiting step. A closer study of the enzymes catalysing these reactions is essential for a better understanding of the microbial activity in the wastewater treatment process. Therefore, development...... of gentle and efficient enzyme extraction methods from environmental samples is very important. In this study we present a method for the extraction of lipases and proteases from activated sludge using the non-ionic detergent Triton X-100, EDTA, and cation exchange resin (CER), alone or in combination...... and negligible protease activity was extracted in the presence of buffer alone, indicating that enzyme extraction was not due to shear force alone. The highest lipase activity was extracted using 0.1% Triton X-100 above which the activity was gradually decreasing. For proteases, the highest activity was obtained...

  8. Seed lipases: sources, applications and properties - a review

    Directory of Open Access Journals (Sweden)

    M. Barros

    2010-03-01

    Full Text Available This paper provides an overview regarding the main aspects of seed lipases, such as the reactions catalyzed, physiological functions, specificities, sources and applications. Lipases are ubiquitous in nature and are produced by several plants, animals and microorganisms. These enzymes exhibit several very interesting features, such as low cost and easy purification, which make their commercial exploitation as industrial enzymes a potentially attractive alternative. The applications of lipases in food, detergents, oils and fats, medicines and fine chemistry, effluent treatment, biodiesel production and in the cellulose pulp industry, as well as the main sources of oilseed and cereal seed lipases, are reviewed.

  9. [Synthesis of biodiesel from crude oil by immobilized lipase].

    Science.gov (United States)

    Li, Junkui; Lu, Jike; Wang, Fang; Tan, Tianwei; Deng, Li

    2009-06-01

    We used immobilized lipase from Candida sp. 99-125 to produce fatty acid methyl esters (FAMEs) from crude oil and methanol. We studied the effects of phospholipids on activity of immobilized lipase, reaction velocity, stability of immobilized lipase and the stability of immobilized lipase in crude and refined oil. Results showed that the activity of the lipase immersed in petroleum ether with 1% phospholipids dropped more quickly than the lipase in petroleum ether without phospholipids. When soybean oil was used without phospholipids as material, the FAMEs yield of 15 min was 26.2%, whereas the yield decreased to 12.4% when there were 5% phospholipids in the soybean oil. However when the phospholipids content was below 1%, the stability of the lipase did not change obviously. The lipase was stable when used to catalyze crude soybean oil and crude jatropha oil, after 10 cycles the FAMEs yield was still above 70%. This lipase showed great potential for industrial production of biodiesel from crude oil.

  10. Esterification of phenolic acids catalyzed by lipases immobilized in organogels.

    Science.gov (United States)

    Zoumpanioti, M; Merianou, E; Karandreas, T; Stamatis, H; Xenakis, A

    2010-10-01

    Lipases from Rhizomucor miehei and Candida antarctica B were immobilized in hydroxypropylmethyl cellulose organogels based on surfactant-free microemulsions consisting of n-hexane, 1-propanol and water. Both lipases kept their catalytic activity, catalyzing the esterification reactions of various phenolic acids including cinnamic acid derivatives. High reaction rates and yields (up to 94%) were obtained when lipase from C. antarctica was used. Kinetic studies have been performed and apparent kinetic constants were determined showing that ester synthesis catalyzed by immobilized lipases occurs via the Michaelis-Menten mechanism.

  11. Immobilization of Lipase by Covalent Binding on Crosslinked Ally Dextran

    Institute of Scientific and Technical Information of China (English)

    WangChen; SongGuoqiang; 等

    1998-01-01

    Lipase was immobilized by covalent binding on crosslinked allyl dextran using SESA as coupling agent.It is shown that this immobilization approach is an efficient one for lipase.The activity of the immobilized lipase can reach to 300-450U/g(dry weight).It exhibits good temperature stability,can retain 88% activity after being incubated at 70℃ for 2h.Special effects will be expected from our immobilized lipase in its applications in organic media due to the nature of the support.

  12. Endothelial lipase is a major determinant of HDL level

    Energy Technology Data Exchange (ETDEWEB)

    Ishida, Tatsuro; Choi, Sungshin; Kundu, Ramendra K.; Hirata, Ken-Ichi; Rubin, Edward M.; Cooper, Allen D.; Quertermous, Thomas

    2003-01-30

    For the past three decades, epidemiologic studies have consistently demonstrated an inverse relationship between plasma HDL cholesterol (HDL-C) concentrations and coronary heart disease (CHD). Population-based studies have provided compelling evidence that low HDL-C levels are a risk factor for CHD, and several clinical interventions that increased plasma levels of HDL-C were associated with a reduction in CHD risk. These findings have stimulated extensive investigation into the determinants of plasma HDL-C levels. Turnover studies using radiolabeled apolipoprotein A-I, the major protein component of HDL, suggest that plasma HDL-C concentrations are highly correlated with the rate of clearance of apolipoprotein AI. However, the metabolic mechanisms by which HDL are catabolized have not been fully defined. Previous studies in humans with genetic deficiency of cholesteryl ester transfer protein, and in mice lacking the scavenger receptor BI (SR-BI), have demonstrated that these proteins participate in the removal of cholesterol from HDL, while observations in individuals with mutations in hepatic lipase indicate that this enzyme hydrolyzes HDL triglycerides. In this issue of the JCI, reports from laboratories of Tom Quertermous and Dan Rader now indicate that endothelial lipase (LIPG), a newly identified member of the lipase family, catalyzes the hydrolysis of HDL phospholipids and facilitates the clearance of HDL from the circulation. Endothelial lipase was initially cloned by both of these laboratories using entirely different strategies. Quertermous and his colleagues identified endothelial lipase as a transcript that was upregulated in cultured human umbilical vein endothelial cells undergoing tube formation, whereas the Rader group cloned endothelial lipase as a transcript that was upregulated in the human macrophage-like cell line THP-1 exposed to oxidized LDL. Database searches revealed that endothelial lipase shows strong sequence similarity to lipoprotein

  13. Comparative and functional genomics of lipases in holometabolous insects.

    Science.gov (United States)

    Horne, Irene; Haritos, Victoria S; Oakeshott, John G

    2009-08-01

    Lipases have key roles in insect lipid acquisition, storage and mobilisation and are also fundamental to many physiological processes underpinning insect reproduction, development, defence from pathogens and oxidative stress, and pheromone signalling. We have screened the recently sequenced genomes of five species from four orders of holometabolous insects, the dipterans Drosophila melanogaster and Anopheles gambiae, the hymenopteran Apis mellifera, the moth Bombyx mori and the beetle Tribolium castaneum, for the six major lipase families that are also found in other organisms. The two most numerous families in the insects, the neutral and acid lipases, are also the main families in mammals, albeit not in Caenorhabditis elegans, plants or microbes. Total numbers of the lipases vary two-fold across the five insect species, from numbers similar to those in mammals up to numbers comparable to those seen in C. elegans. Whilst there is a high degree of orthology with mammalian lipases in the other four families, the great majority of the insect neutral and acid lipases have arisen since the insect orders themselves diverged. Intriguingly, about 10% of the insect neutral and acid lipases have lost motifs critical for catalytic function. Examination of the length of lid and loop regions of the neutral lipase sequences suggest that most of the insect lipases lack triacylglycerol (TAG) hydrolysis activity, although the acid lipases all have intact cap domains required for TAG hydrolysis. We have also reviewed the sequence databases and scientific literature for insights into the expression profiles and functions of the insect neutral and acid lipases and the orthologues of the mammalian adipose triglyceride lipase which has a pivotal role in lipid mobilisation. These data suggest that some of the acid and neutral lipase diversity may be due to a requirement for rapid accumulation of dietary lipids. The different roles required of lipases at the four discrete life stages of

  14. Lipoprotein lipase deficiency with visceral xanthomas

    Energy Technology Data Exchange (ETDEWEB)

    Servaes, Sabah; Bellah, Richard [Department of Radiology, Philadelphia, PA (United States); Verma, Ritu [Department of Gastroenterology, Philadelphia, PA (United States); Pawel, Bruce [Department of Pathology, Philadelphia, PA (United States)

    2010-08-15

    Lipoprotein lipase deficiency (LLD) is a rare metabolic disorder that typically presents with skin xanthomas and pancreatitis in childhood. We report a case of LLD in an infant who presented with jaundice caused by a pancreatic head mass. Abdominal imaging also incidentally revealed hyperechoic renal masses caused by renal xanthomas. This appearance of the multiple abdominal masses makes this a unique infantile presentation of LLD. (orig.)

  15. Characterization of Cross-Linked Lipase Aggregates

    DEFF Research Database (Denmark)

    Prabhavathi Devi, Bethala Lakshmi Anu; Guo, Zheng; Xu, Xuebing

    2009-01-01

    Commercially available microbial lipases from different sources were immobilized as cross-linked enzyme aggregates (CLEAs) using different precipitants and glutaraldehyde as cross-linkers. These CLEAs were assayed based on esterification between lauric acid and n-propanol in solvent-free systems...... change upon CLEA formation. This work presents a characterization of CLEAs based on an esterification activity assay, which is useful for exploring the synthetic application potential of CLEA technology with favorable perspectives....

  16. Immobilization of lipase from Fusarium solani FS1 Imobilização de lipase de Fusarium solani FS1

    Directory of Open Access Journals (Sweden)

    Kirsty Knight

    2000-09-01

    Full Text Available Lipase from Fusarium solani FS1 was immobilized by covalent attachment to polyacrylamide beads and onto magnetized Dacron, retaining 12% and 97% of activity, respectively. Lipase was also entrapped within polyacrylamide beads, retaining 53% of activity. Investigations of the kinetic characteristics of the immobilized derivatives using triolein as substrate showed that lipase immobilized onto polyacrilamide beads and Dacron did not follow Michaelis-Menten kinetics.Lipase de Fusarium solani FS1 foi imobilizada por ligação covalente usando esferas de poliacrilamida e Dacron magnetizado, retendo 12%, e 97% de atividade, respectivamente. A lipase foi também enclausurada em esferas de poliacrilamida e reteve 53% de sua atividade específica. Investigações sobre o comportamento cinético usando trioleína como substrato mostraram que as lipases imobilizadas não seguem a cinética de Michaelis-Menten.

  17. Biodiesel production by transesterification using immobilized lipase.

    Science.gov (United States)

    Narwal, Sunil Kumar; Gupta, Reena

    2013-04-01

    Biodiesel can be produced by transesterification of vegetable or waste oil catalysed by lipases. Biodiesel is an alternative energy source to conventional fuel. It combines environmental friendliness with biodegradability, low toxicity and renewability. Biodiesel transesterification reactions can be broadly classified into two categories: chemical and enzymatic. The production of biodiesel using the enzymatic route eliminates the reactions catalysed under acid or alkali conditions by yielding product of very high purity. The modification of lipases can improve their stability, activity and tolerance to alcohol. The cost of lipases and the relatively slower reaction rate remain the major obstacles for enzymatic production of biodiesel. However, this problem can be solved by immobilizing the enzyme on a suitable matrix or support, which increases the chances of re-usability. The main factors affecting biodiesel production are composition of fatty acids, catalyst, solvents, molar ratio of alcohol and oil, temperature, water content, type of alcohol and reactor configuration. Optimization of these parameters is necessary to reduce the cost of biodiesel production.

  18. Lipase production by recombinant strains of Aspergillus niger expressing a lipase-encoding gene from Thermomyces lanuginosus

    DEFF Research Database (Denmark)

    Prathumpai, Wai; Flitter, S.J.; Mcintyre, Mhairi

    2004-01-01

    Two recombinant strains of Aspergillus niger (NW 297-14 and NW297-24) producing a heterologous lipase from Thermomyces lanuginosus were constructed. The heterologous lipase was expressed using the TAKA amylase promoter from Aspergillus oryzae. The production kinetics of the two strains on different...... shows that it is possible to obtain high productivities of heterologous fungal enzymes in A. niger. However, SDS-PAGE analysis showed that most of the produced lipase was bound to the cell wall....

  19. Immobilization of a Commercial Lipase from Penicillium camembertii (Lipase G by Different Strategies

    Directory of Open Access Journals (Sweden)

    Adriano A. Mendes

    2011-01-01

    Full Text Available The objective of this work was to select the most suitable procedure to immobilize lipase from Penicillium camembertii (Lipase G. Different techniques and supports were evaluated, including physical adsorption on hydrophobic supports octyl-agarose, poly(hydroxybutyrate and Amberlite resin XAD-4; ionic adsorption on the anionic exchange resin MANAE-agarose and covalent attachment on glyoxyl-agarose, MANAE-agarose cross-linked with glutaraldehyde, MANAE-agarose-glutaraldehyde, and epoxy-silica-polyvinyl alcohol composite. Among the tested protocols, the highest hydrolytic activity (128.2 ± 8.10 IU·g−1 of support was achieved when the lipase was immobilized on epoxy-SiO2-PVA using hexane as coupling medium. Lipase immobilized by ionic adsorption on MANAE-agarose also gave satisfactory result, attaining 55.6 ± 2.60 IU·g−1 of support. In this procedure, the maximum loading of immobilized enzyme was 9.3 mg·g−1 of gel, and the highest activity (68.8 ± 2.70 IU·g−1 of support was obtained when 20 mg of protein·g−1 was offered. Immobilization carried out in aqueous medium by physical adsorption on hydrophobic supports and covalent attachment on MANAE-agarose-glutaraldehyde and glyoxyl-agarose was shown to be unfeasible for Lipase G. Thermal stability tests revealed that the immobilized derivative on epoxy-SiO2-PVA composite using hexane as coupling medium had a slight higher thermal stability than the free lipase.

  20. Lipase - Catalyzed glycerolysis of sunflower oil to produce partial glycerides.

    Directory of Open Access Journals (Sweden)

    Zaher, F. A.

    1998-12-01

    Full Text Available Partial glycerides were prepared by glycerolysis of sunflower oil in presence of lipase enzyme as catalyst. Six lipases of different origins were used and compared for their catalytic activity. These include Chromobacterium lipase, pancreatic lipase, Rhizopus arrhizus lipase, lyophilized lipase (plant lipase in addition to two lipase preparations derived from Rhizopus japonicas; Lilipase A-10 and Lilipase B-2. Chromobacterium lipase was found to be the most active as glycerolysis catalyst whereas lyophilized lipase; a plant preparation from wheat germ was the least active. The results have also shown that the lipase type affects also the product polarity and hence its field of application as a food emulsifier. Less polar products can be obtained using Chromobacterium lipase whereas the more polar ones using a fungal lipase preparation «Lipase A-10». The product polarity is also influenced by the process temperature but the mode of its effect is different for different lipases.

    Se prepararon glicéridos parciales mediante glicerolisis de aceite de girasol en presencia de lipasa como catalizador. Seis lipasas de orígenes diferentes se utilizaron y compararon en función de su actividad catalítica. Estas incluyeron lipasa de Chromobacterium, lipasa pancreática, lipasa de Rhizopus arrhizus, lipasa liofilizada (lipasa vegetal además de dos preparaciones de lipasa derivadas de Rhizopus japonicus: lilipase A-10 y lilipase B-2. Se encontró que la lipasa de Chromobacterium fue la más activa como catalizador en la glicerolisis mientras que la lipasa liofilizada, preparación vegetal a partir de germen de trigo, fue la menos activa. Los resultados mostraron que los tipos de lipasa afectan también a la polaridad de los productos y por tanto a los rendimientos en su aplicación como emulsificantes alimentarios. Los productos menos polares pueden obtenerse usando lipasa de

  1. Pancreatic lipase-related protein 2 digests fats in human milk and formula in concert with gastric lipase and carboxyl ester lipase

    Science.gov (United States)

    Johnson, Karin; Ross, Leah; Miller, Rita; Xiao, Xunjun; Lowe, Mark E.

    2013-01-01

    INTRODUCTION Dietary fats must be digested into fatty acids and monoacylglycerols prior to absorption. In adults, colipase-dependent pancreatic triglyceride lipase (PTL) contributes significantly to fat digestion. In newborn rodents and humans, the pancreas expresses low levels of PTL. In rodents, a homologue of PTL, pancreatic lipase related protein 2 (PLRP2) and carboxyl ester lipase (CEL) compensate for the lack of PTL. In human newborns, the role for PLRP2 in dietary fat digestion is unclear. To clarify the potential of human PLRP2 to influence dietary fat digestion in newborns, we determined PLRP2 activity against human milk and infant formula. METHODS The activity of purified recombinant PLRP2, gastric lipase and CEL against fats in human milk and formula was measured with each lipase alone and in combination with a standard pH-stat assay. RESULTS Colipase added to human milk stimulated fat digestion. PLRP2 and CEL had activity against human milk and formula. Pre-digestion with gastric lipase increased PLRP2 activity against both substrates. Together, CEL and PLRP2 activity was additive with formula and synergistic with human milk. CONCLUSIONS PLRP2 can digest fats in human milk and formula. PLRP2 acts in concert with CEL and gastric lipase to digest fats in human milk in vitro. PMID:23732775

  2. Long-Term Follow-Up of HLA-A2+ Patients with High-Risk, Hormone-Sensitive Prostate Cancer Vaccinated with the Prostate Specific Antigen Peptide Homologue (PSA146-154

    Directory of Open Access Journals (Sweden)

    Supriya Perambakam

    2010-01-01

    Full Text Available Twenty-eight HLA-A2+ patients with high-risk, locally advanced or metastatic, hormone-sensitive prostate cancer were immunized with a peptide homologue of prostate-specific antigen, PSA146-154, between July 2002 and September 2004 and monitored for clinical and immune responses. Fifty percent of the patients developed strong PSA146-154-peptide-specific delayed-type hypersensitivity skin responses, tetramer and/or IFN-γ responses within one year. Thirteen patients had stable or declining serum levels of PSA one year post-vaccination. A decreased risk of biochemical progression was observed in patients who developed augmented tetramer responses at six months compared to pre-vaccination levels (P=.02. Thirteen patients have died while 15 patients remain alive with a mean overall survival of 60 months (95% CI, 51 to 68 months per Kaplan-Meier analysis. A trend towards greater overall survival was detected in men with high-risk, hormone-sensitive CaP who developed specific T-cell immunity following vaccination with PSA146-154 peptide.

  3. Pycnogenol supplementation promotes lipolysis via activation of cAMP-dependent PKA in ob/ob mice and primary-cultured adipocytes.

    Science.gov (United States)

    Ho, Jin-Nyoung; Kim, Ok-Kyung; Nam, Da-Eun; Jun, Woojin; Lee, Jeongmin

    2014-01-01

    This study investigated the PKA-dependent inhibitory effect of pycnogenol (Pyc) on lipolysis using ob/ob mice and primary mouse adipocytes. Supplementation of Pyc at 30 mg/kg significantly reduced body weight gain and visceral fat mass. The serum and hepatic triglyceride (TG) and total cholesterol (TC) levels were reduced by Pyc supplementation, and high density lipoprotein (HDL)-cholesterol level significantly increased. In addition, hormone sensitive lipase (HSL) and adipose triglyceride lipase (ATGL) mRNA levels increased with Pyc supplementation in adipose tissue of ob/ob mice. The treatment of primary cultured adipocytes with Pyc at 100 μg/mL significantly increased glycerol release, cAMP level by reduction of phosphodiestersae-3B (PDE3B), and HSL levels, but decreased protein levels of perilipin A and fatty acid synthetase (FAS). The PKA inhibitor (H89) clearly blocked the cellular levels of perilipin A and HSL, suggesting that Pyc promotes lipolysis of adipocytes through activation of cAMP-dependent PKA, resulting in induction of HSL and reduction of perilipin A. Therefore, this study may elucidate the possible mechanism of Pyc, which is a candidate for weight loss through stimulation of lipolysis.

  4. The AIS-HSL Optimization:An Artificial Immune System with Heuristic Social Learning#%一种带有启发式社会学习机制的人工免疫系统

    Institute of Scientific and Technical Information of China (English)

    李中华; 贺春辉

    2012-01-01

    This paper proposes an artificial immune system with heuristic social learning (AIS-HSL) for optimization. In the AIS-HSL optimization, the candidate antibodies is separated into two swarms, i.e., the elitist swarm (ES) and the common swarm (CS). Different swarms experience different mutation processes, i.e. , a self-learning strategy is used for the ES while a heuristic social-learning (HSL) mechanism is applied to the CS. In the HSL mechanism, each antibody in CS learns from an selected antibody in ES based on the probability determined by the affinity to avoid falling into the local optima. Some comparative numerical simulations are arranged to evaluate the performance of the proposed AIS-HSL. The results demonstrate that the proposed AIS-HSL outperforms the canonical opt-aiNet, the IA-AIS and the AAIS-2S in convergence speed and solution accuracy.%本文提出一种带有启发式社会学习机制的人工免疫系统优化算法(AIS-HSL)。在AIS-HSL中,候选抗体被分成两个群,即精英群(ES)和普通群(CS)。不同的群执行不同的变异机制,即精英群采用自学习机制,普通群采用启发式社会学习(HSL)机制。在HSL机制中,CS中的每个抗体根据亲和力依概率选择ES中的抗体进行学习,以避免陷入局部最优。通过一些比较性实验,来评估 AIS-HSL 算法的性能。实验结果表明,与传统的opt-aiNet算法和IA-AIS算法相比,本文提出的AIS-HSL算法有着更高的收敛精度和收敛速度。

  5. Properties of an immobilized lipase of Bacillus coagulans BTS-1.

    Science.gov (United States)

    Kanwari, S S; Srivastava, M; Chimni, S S; Ghazi, I A; Kaushal, R K; Joshi, G K

    2004-01-01

    Lipase (EC 3.1.1.3) is a tri-acylglycerol ester hydrolase, catalysing the hydrolysis of tri-, di-, and mono-acylglycerols to glycerol and fatty acids. To study the effect of adsorption of a lipase obtained from Bacillus coagulans BTS-1, its lipase was immobilized on native and activated (alkylated) matrices, i.e. silica and celite. The effect of pH, temperature, detergents, substrates, alcohols, organic solvent etc. on the stability of the immobilized enzyme was evaluated. The gluteraldahyde or formaldehyde (at 1% and 2% concentration, v/v) activated matrix was exposed to the Tris buffered lipase. The enzyme was adsorbed/entrapped more rapidly on to the activated silica than on the activated celite. The immobilized lipase showed optimal activity at 50 degrees C following one-hour incubation. The lipase was specifically more hydrolytic to the medium C-length ester (p-nitro phenyl caprylate than p-nitro phenyl laurate). The immobilization/entrapment enhanced the stability of the lipase at a relatively higher temperature (50 degrees C) and also promoted enzyme activity at an acidic pH (pH 5.5). Moreover, the immobilized lipase was quite resistant to the denaturing effect of SDS.

  6. pH-optima in lipase-catalysed esterification

    NARCIS (Netherlands)

    Buthe, Andreas; Recker, Tobias; Heinemann, Matthias; Hartmeier, Winfried; Büchs, Jochen; Ansorge-Schumacher, Marion B.

    2005-01-01

    Though lipases are frequently applied in ester synthesis, fundamental information on optimal pH or substrate concentration, can almost only be found for the reverse reaction - hydrolysis. This study demonstrates that the pH-optima of lipase-catalysed esterifications differ significantly from the opt

  7. pH-optima in lipase-catalysed esterification

    NARCIS (Netherlands)

    Buthe, Andreas; Recker, Tobias; Heinemann, Matthias; Hartmeier, Winfried; Büchs, Jochen; Ansorge-Schumacher, Marion B.

    2005-01-01

    Though lipases are frequently applied in ester synthesis, fundamental information on optimal pH or substrate concentration, can almost only be found for the reverse reaction - hydrolysis. This study demonstrates that the pH-optima of lipase-catalysed esterifications differ significantly from the

  8. Surfactant-activated lipase hybrid nanoflowers with enhanced enzymatic performance

    Science.gov (United States)

    Cui, Jiandong; Zhao, Yamin; Liu, Ronglin; Zhong, Cheng; Jia, Shiru

    2016-01-01

    Increasing numbers of materials have been extensively used as platforms for enzyme immobilization to improve catalytic performance. However, activity of the most of the enzymes was declined after immobilization. Here, we develop a surfactant-activated lipase-inorganic flowerlike hybrid nanomaterials with rational design based on interfacial activation and self-assembly. The resulting surfactant-activated lipase-inorganic hybird nanoflower (activated hNF-lipase) exhibited 460% and 200% higher activity than native lipase and conventional lipase-inorganic hybird nanoflower (hNF-lipase). Furthermore, the activated hNF-lipase displayed good reusability due to its monodispersity and mechanical properties, and had excellent long-time stability. The superior catalytic performances were attributed to both the conformational modulation of surfactants and hierarchical structure of nanoflowers, which not only anchored lipases in an active form, but also decreased the enzyme-support negative interaction and mass-transfer limitations. This new biocatalytic system is promising to find widespread use in applications related to biomedicine, biosensor, and biodiesel. PMID:27297609

  9. Lipase-Catalyzed Modification of Canola Oil with Caprylic Acid

    DEFF Research Database (Denmark)

    Wang, Yingyao; Luan, Xia; Xu, Xuebing

    Lipase-catalyzed acidolysis of canola oil with caprylic acid was performed to produce structured lipids. Six commercial lipases from different sources were screened for their ability to incorporate the caprylic acid into the canola oil. The positional distribution of FA on the glycerol backbone o...

  10. THE EFFECT OF LIPASE INGESTION UPON BLOOD LIPID LEVELS.

    Science.gov (United States)

    levels. The optical density and hence lipid levels of the blood plasmas were lowered in all subjects when sufficient lipase was ingested . The total...obtain this effect. The blood cholesterol values of normal subjects were not affected, nor was that of a hyperlipemic subject who ingested the lipase

  11. Characteristics of lipase isolated from coconut (Cocos nucifera linn ...

    African Journals Online (AJOL)

    GREGO

    2007-03-19

    Mar 19, 2007 ... Lipase from coconut plant grown under complete nutrient ... exceptionally high lipolytic activity probably in order to ... in chocolate crumb, in improvement of egg white .... fat. Berner and Hammond (1970) found that the lipase from oat .... isolation and purification of total lipids from animal tissues J. Biol. Chem ...

  12. Papaya (Carica papaya) lipase with some distinct acyl and alkyl specificities as compared with microbial lipases.

    Science.gov (United States)

    Gandhi, N N; Mukherjee, K D

    2000-12-01

    Lipase from papaya (Carica papaya) latex (CPL), Candida antarctica lipase B (Novozym 435, NOV) and Rhizomucor miehei lipase (Lipozyme IM 20, LIP) were used as biocatalysts for the esterification of caprylic acid with straight-chain saturated C(4)-C(18) alcohols and unsaturated C(18) alcohols, such as cis-9-octadecenyl (oleyl, C(18:1), n-9), cis-6-octadecenyl (petroselinyl, C(18:1), n-12), cis-9,cis-12-octadecadienyl (linoleyl, C(18:2), n-6), all-cis-9,12,15-octadecatrienyl (alpha-linolenyl, C(18:3), n-3) and all-cis-6,9,12-octadecatrienyl (gamma-linolenyl, C(18:3), n-6) alcohols. With CPL, highest activity was found in the esterification of octanol and decanol, whereas both NOV and LIP showed a broad chain-length-specificity for the alcohols. CPL, as opposed to the microbial lipases, strongly discriminated against all the saturated long-chain ( > C(12)) and unsaturated C(18) alcohols.

  13. Activity and stability of immobilized lipases in lipase-catalyzed modification of peanut oil

    Directory of Open Access Journals (Sweden)

    Soumanou Mohamed M.

    2004-11-01

    Full Text Available Fatty acid release during lipolysis of peanut oil using microbial free and immobilized lipases in aqueous media was developed. Immobilized lipase from Rhizomucor miehei (RML gave the best result from its ability to clive different fatty acids from peanut oil in such media. In organic solvent, interesterification of peanut oil with tricaprylin using immobilized lipases from RML, Chromobacterium viscosum (CVL and Candida rugosa (CRL was performed. The best substrate molar ratio of tricaprylin to peanut oil found was in the range 0.7 to 0.8. Using substrate molar ratio 0.7, high amount of structured triglyceride ST (about 35% MLM, 44% LML triglyceride fractions was obtained with lipase from RML in n-hexane. The results found in solvent free system were in some cases quite similar to that obtained in organic solvent. In nine successive batch interesterification in solvent free medium using immobilized RML and CRL, no significant loss of amount of both produced triacylglycerol fractions until batch 7 was observed with RML.

  14. Process Technology for Immobilized LipaseProcess Technology for Immobilized Lipase-catalyzed

    DEFF Research Database (Denmark)

    Xu, Yuan

    evaluation has been performed for six processes composed of transesterification and product purification for making ‘in-spec’ biodiesel and the conventional chemical process is taken as a bench mark for comparison. The optimal process is a process composed of lipase-catalyzed transesterification with ‘in...

  15. Kinetic model of biodiesel production using immobilized lipase Candida antarctica lipase B

    DEFF Research Database (Denmark)

    Fedosov, Sergey; Brask, Jesper; Pedersen, Anders K.

    2013-01-01

    We have designed a kinetic model of biodiesel production using Novozym 435 (Nz435) with immobilized Candida antarctica lipase B (CALB) as a catalyst. The scheme assumed reversibility of all reaction steps and imitated phase effects by introducing various molecular species of water and methanol...

  16. Endothelial and lipoprotein lipases in human and mouse placenta

    DEFF Research Database (Denmark)

    Lindegaard, Marie L S; Olivecrona, Gunilla; Christoffersen, Christina;

    2005-01-01

    Placenta expresses various lipase activities. However, a detailed characterization of the involved genes and proteins is lacking. In this study, we compared the expression of endothelial lipase (EL) and LPL in human term placenta. When placental protein extracts were separated by heparin......-Sepharose affinity chromatography, the EL protein eluted as a single peak without detectable phospholipid or triglyceride (TG) lipase activity. The major portion of LPL protein eluted slightly after EL. This peak also had no lipase activity and most likely contained monomeric LPL. Fractions eluting at a higher Na......Cl concentration contained small amounts of LPL protein (most likely dimeric LPL) and had substantial TG lipase activity. In situ hybridization studies showed EL mRNA expression in syncytiotrophoblasts and endothelial cells and LPL mRNA in syncytiotrophoblasts. In contrast, immunohistochemistry showed EL and LPL...

  17. Applications of immobilized Thermomyces lanuginosa lipase in interesterification

    DEFF Research Database (Denmark)

    Yang, Tiankui; Fruekilde, Maj-Britt; Xu, Xuebing

    2003-01-01

    The aim of this work was to investigate the catalytic functions of a new immobilized Thermomyces lanuginosa lipase in interesterification and to optimize the conditions of interesterification for the production of human milk fat substitutes (HMFS) containing n-3 PUFA by response surface methodology...... (RSM). Thermomyces lanuginosa lipase had an activity similar to that of immobilized Rhizomucor miehei lipase (Lipozyme RM IM) in the glycerolysis of sunflower oil, but the former had higher activity at a low reaction temperature (5degreesC). Thermomyces lanuginosa lipase was found to have much lower...... catalytic activity than Lipozyme RM IM in the acidolysis of sunflower oil with caprylic acid. However, the activity of T. lanuginosa lipase was only slightly lower than that of Lipozyme RM IM in the ester-ester exchange between tripalmitin (PPP) and the ethyl esters of EPA and DHA (EE). For this reason...

  18. Microwave-assisted rapid characterization of lipase selectivities.

    Science.gov (United States)

    Bradoo, Sapna; Rathi, Pooja; Saxena, R K; Gupta, Rani

    2002-04-18

    A rapid screening procedure for characterization of lipase selectivities using microwaves was developed. The rate of reaction of various commercial lipases (porcine pancreas, Mucor miehei, Candida rugosa, Pseudomonas cepacia) as well as lipases from laboratory isolates-Bacillus stearothermophilus and Burkholderia cepacia RGP-10 for triolein hydrolysis was 7- to 12-fold higher in a microwave oven as compared to that by pH stat. The esterification of sucrose/methanol and ascorbic acid with different fatty acids was also achieved within 30 s in a microwave using porcine pancreas, B. stearothermophilus SB-1 and B. cepacia RGP-10 lipases. The relative rates and selectivity of the lipases both for hydrolytic and synthesis reactions remains unaltered. However, the rate of reaction was dynamically enhanced when exposed to microwaves. Microwave-assisted enzyme catalysis can become an attractive procedure for rapid characterization of large number of enzyme samples and substrates, which otherwise is a cumbersome and time-consuming exercise.

  19. Monoolein production by triglycerides hydrolysis using immobilized Rhizopus oryzae lipase.

    Science.gov (United States)

    Ghattas, Nesrine; Abidi, Ferid; Galai, Said; Marzouki, M Nejib; Salah, Abderraouf Ben

    2014-07-01

    Lipase extracted from Rhizopus oryzae was immobilized in alginate gel beads. The effects of the immobilization conditions, such as, alginate concentration, CaCl2 concentration and amount of initial enzyme on retained activity (specific activity ratio of entrapped active lipase to free lipase) were investigated. The optimal conditions for lipase entrapment were determined: 2% (w/v) alginate concentration, 100mM CaCl2 and enzyme ratio of 2000IU/mL.In such conditions, immobilized lipase by inclusion in alginate showed a highest stability and activity, on olive oil hydrolysis reaction where it could be reused for 10 cycles. After 15min of hydrolysis reaction, the mass composition of monoolein, diolein and triolein were about 78%, 10% and 12%. Hydrolysis' products purification by column chromatography lead to a successful separation of reaction compounds and provide a pure fraction of monoolein which is considered as the widest used emulsifier in food and pharmaceutical industries.

  20. Inhibitory activity of benzophenones from Anemarrhena asphodeloides on pancreatic lipase.

    Science.gov (United States)

    Jo, Yang Hee; Kim, Seon Beom; Ahn, Jong Hoon; Liu, Qing; Hwang, Bang Yeon; Lee, Mi Kyeong

    2013-04-01

    Pancreatic lipase is a key enzyme for lipid absorption by hydrolysis of total dietary fats. Therefore, inhibition of pancreatic lipase is suggested to be an effective therapy in the regulation of obesity. The EtOAc-soluble fraction of Anemarrhena asphodeloides rhizomes significantly inhibited pancreatic lipase activity as assessed using porcine pancreatic lipase as an in vitro assay system. Further fractionation of the EtOAc-soluble fraction of A. asphodeloides led to the isolation of a new benzophenone glycoside, zimoside A (1), together with the eleven known compounds iriflophenone (2), 2,4',6-trihydroxy-4-methoxybenzophenone (3), foliamangiferoside A (4), (2,3-dihydroxy-4-methoxyphenyl)(4-hydroxyphenyl)-methanone (5), 1,4,5,6,-tetrahydroxyxanthone (6), isosakuranetin (7), 4-hydroxybenzoic acid (8), 4-hydroxyacetophenone (9), vanillic acid (10), tyrosol (11) and 5-hydroxymethyl-2-furaldehyde (12). Among the isolated compounds, 3, 5 and 10 showed significant inhibition of pancreatic lipase activity.

  1. Bacterial lipases: A review on purification and characterization.

    Science.gov (United States)

    Javed, Saira; Azeem, Farrukh; Hussain, Sabir; Rasul, Ijaz; Siddique, Muhammad Hussnain; Riaz, Muhammad; Afzal, Muhammad; Kouser, Ambreen; Nadeem, Habibullah

    2017-08-01

    Lipase (E.C.3.1.1.3) belongs to the hydrolases and is also known as fat splitting, glycerol ester hydrolase or triacylglycerol acylhydrolase. Lipase catalyzes the hydrolysis of triglycerides converting them to glycerol and fatty acids in an oil-water interface. These are widely used in food, dairy, flavor, pharmaceuticals, biofuels, leather, cosmetics, detergent, and chemical industries. Lipases are of plant, animal, and microbial origin, but microbial lipases are produced at industrial level and represent the most widely used class of enzymes in biotechnological applications and organic chemistry. Phylogenetic analysis and comparison of residues around GxSxG motif provided an insight to the diversity among bacterial lipases. A variety of para-Nitrophenyl (p-NP) esters having C2 to C16 (p-NP acetate to p-NP palmitate) in their fatty acid side chain can be hydrolyzed by bacterial lipases. Large heterogeneity has been observed in molecular and catalytic characteristics of lipases including molecular mass; 19-96 kDa, Km; 0.0064-16.58 mM, Kcat; 0.1665-1.0 × 10(4) s(-1) and Kcat/Km; 26.02-7377 s(-1)/mM. Optimal conditions of their working temperature and pH have been stated 15-70 °C and 5.0-10.8, respectively and are strongly associated with the type and growth conditions of bacteria. Surface hydrophobicity, enzyme activity, stability in organic solvents and at high temperature, proteolytic resistance and substrate tolerance are the properties of bacterial lipases that have been improved by engineering. Bacterial lipases have been extensively studied during last decade. However, their wider applications demand a detailed review on purification, catalytic characterization and applications of lipases. Copyright © 2017. Published by Elsevier Ltd.

  2. La lipase de Candida rugosa : caractérisation biochimique

    Directory of Open Access Journals (Sweden)

    Mtibaa Hounaida

    2002-01-01

    Full Text Available Les lipases ou triacylglycérols hydrolases (EC 3.1.1.3 sont des enzymes qui agissent en milieu hétérogène. Ces enzymes catalysent l’hydrolyse des liaisons esters des triacylglycérols à l’interface huile/eau [1]. Leur particularité vient du fait que ces enzymes sont plus actives sur les lipides qui sont sous forme agrégée [2]. Les lipases sont présentes dans la plupart des tissus animaux et végétaux ainsi que chez les microorganismes qui constituent une source importante de production de lipases à grande échelle. À ce jour, de nombreuses lipases de microorganismes ont été purifiées et caractérisées et certaines d’entres elles ont été cristallisées (lipase de Pseudomonas glumae [3], lipase de Rhizomucor miehei [4], lipase Geotrichum candidum [5], lipase de Candida rugosa [6].... La lipase de Candida cylindracea (qui est l’ancien nom de Candida rugosa a été cristallisée en présence et en l’absence d’inhibiteurs [7]. Il s’agit d’une alpha/beta hydrolase comprenant 11 brins beta entourés par 8 hélices alpha [6]. La triade catalytique est cachée sous un flap constitué de 26 résidus d’aminoacides. Dans le présent travail, nous avons cherché à étudier quelques caractéristiques biochimiques de la lipase de Candida rugosa (CRL qui a été purifiée dans notre laboratoire à partir de la poudre commercialisée.

  3. Mechanism of acetaldehyde-induced deactivation of microbial lipases

    Directory of Open Access Journals (Sweden)

    Jaeger Karl E

    2011-02-01

    Full Text Available Abstract Background Microbial lipases represent the most important class of biocatalysts used for a wealth of applications in organic synthesis. An often applied reaction is the lipase-catalyzed transesterification of vinyl esters and alcohols resulting in the formation of acetaldehyde which is known to deactivate microbial lipases, presumably by structural changes caused by initial Schiff-base formation at solvent accessible lysine residues. Previous studies showed that several lipases were sensitive toward acetaldehyde deactivation whereas others were insensitive; however, a general explanation of the acetaldehyde-induced inactivation mechanism is missing. Results Based on five microbial lipases from Candida rugosa, Rhizopus oryzae, Pseudomonas fluorescens and Bacillus subtilis we demonstrate that the protonation state of lysine ε-amino groups is decisive for their sensitivity toward acetaldehyde. Analysis of the diverse modification products of Bacillus subtilis lipases in the presence of acetaldehyde revealed several stable products such as α,β-unsaturated polyenals, which result from base and/or amino acid catalyzed aldol condensation of acetaldehyde. Our studies indicate that these products induce the formation of stable Michael-adducts at solvent-accessible amino acids and thus lead to enzyme deactivation. Further, our results indicate Schiff-base formation with acetaldehyde to be involved in crosslinking of lipase molecules. Conclusions Differences in stability observed with various commercially available microbial lipases most probably result from different purification procedures carried out by the respective manufacturers. We observed that the pH of the buffer used prior to lyophilization of the enzyme sample is of utmost importance. The mechanism of acetaldehyde-induced deactivation of microbial lipases involves the generation of α,β-unsaturated polyenals from acetaldehyde which subsequently form stable Michael-adducts with the

  4. A novel organic solvent tolerant lipase from Bacillus sphaericus 205y: extracellular expression of a novel OST-lipase gene.

    Science.gov (United States)

    Sulong, Moohamad Ropaning; Abdul Rahman, Raja Noor Zaliha Raja; Salleh, Abu Bakar; Basri, Mahiran

    2006-10-01

    An organic solvent tolerant (OST) lipase gene from Bacillus sphaericus 205y was successfully expressed extracellularly. The expressed lipase was purified using two steps purification; ultrafiltration and hydrophobic interaction chromatography (HIC) to 8-fold purity and 32% recovery. The purified 205y lipase revealed homogeneity on denaturing gel electrophoresis and the molecular mass was at approximately 30 kDa. The optimum pH for the purified 205y lipase was 7.0-8.0 and its stability showed a broad range of pH value between pH 5.0 to 13.0 at 37 degrees C. The purified 205y lipase exhibited an optimum temperature of 55 degrees C. The activity of the purified lipase was stimulated in the presence of Ca2+ and Mg2+. Ethylenediaminetetraacetic acid (EDTA) has no effect on its activity; however inhibition was observed with phenylmethane sulfonoyl fluoride (PMSF) a serine hydrolase inhibitor. Organic solvents such as dimethylsulfoxide (DMSO), methanol, p-xylene and n-decane enhanced the activity. Studies on the effect of oil showed that the lipase was most active in the presence of tricaprin (C10). The lipase exhibited 1,3 positional specificity. Bacter

  5. Role of Hepatic Lipase and Endothelial Lipase in High-Density Lipoprotein-Mediated Reverse Cholesterol Transport

    NARCIS (Netherlands)

    Annema, Wijtske; Tietge, Uwe J. F.

    2011-01-01

    Reverse cholesterol transport (RCT) constitutes a key part of the atheroprotective properties of high-density lipoproteins (HDL). Hepatic lipase (HL) and endothelial lipase (EL) are negative regulators of plasma HDL cholesterol levels. Although overexpression of EL decreases overall macrophage-to-fe

  6. Enzymatic interesterification of palm stearin and coconut oil by a dual lipase system

    DEFF Research Database (Denmark)

    Ibrahim, Nuzul Amri Bin; Guo, Zheng; Xu, Xuebing

    2008-01-01

    Enzymatic interesterification of palm stearin with coconut oil was conducted by applying a dual lipase system in comparison with individual lipase-catalyzed reactions. The results indicated that a synergistic effect occurred for many lipase combinations, but largely depending on the lipase species...

  7. Interesterification of Milk Fat with Oleic Acid Catalyzed by Immobilized Rhizopus oryzae Lipase

    NARCIS (Netherlands)

    OBA, T; Witholt, B.

    1994-01-01

    Milk fat was interesterified with oleic acid by catalysis of an immobilized lipase in a microaqueous two-phase system. A commercial lipase from Rhizopus oryzae and a controlled pore glass carrier were selected for preparation of an immobilized lipase. The prepared immobilized lipase showed a Michael

  8. INTERESTERIFICATION OF MILK-FAT WITH OLEIC-ACID CATALYZED BY IMMOBILIZED RHIZOPUS-ORYZAE LIPASE

    NARCIS (Netherlands)

    OBA, T; WITHOLT, B

    1994-01-01

    Milk fat was interesterified with oleic acid by catalysis of an immobilized lipase in a microaqueous two-phase system. A commercial lipase from Rhizopus oryzae and a controlled pore glass carrier were selected for preparation of an immobilized lipase. The prepared immobilized lipase showed a Michael

  9. Lipases as Tools in the Synthesis of Prodrugs from Racemic 9-(2,3-Dihydroxypropyladenine

    Directory of Open Access Journals (Sweden)

    Marcela Krečmerová

    2012-11-01

    Full Text Available Lipases from Geotrichum candidum 4013 (extracellular lipase and cell-bound lipase were immobilized by adsorption on chitosan beads. The enzyme preparations were tested in the synthesis of ester prodrugs from racemic 9-(2,3-dihydroxypropyladenine in dimethylformamide with different vinyl esters (acetate, butyrate, decanoate, laurate, palmitate. The transesterification activities of these immobilized enzymes were compared with commercially available lipases (lipase from hog pancreas, Aspergillus niger, Candida antarctica, Pseudomonas fluorescens. Lipase from Candida antarctica was found to be the most efficient enzyme regarding chemical yield of the desired products, while transesterification by lipase from Aspergillus niger resulted in lower yields.

  10. Wnt/β-catenin signaling pathway and lipolysis enzymes participate in methylprednisolone induced fat differential distribution between subcutaneous and visceral adipose tissue.

    Science.gov (United States)

    Xiao, Xinhua; Li, Han; Yang, Jiaojiao; Qi, Xiaoyan; Zu, Xuyu; Yang, Jing; Zhong, Jing; Cao, Renxian; Liu, Jianghua; Wen, Gebo

    2014-06-01

    Glucocorticoids (GCs) are well known to induce fat distribution, which is consistent with the central adiposity phenotype seen in Cushing's syndrome. GCs have been proposed to be both adipogenic and lipolytic in action within adipose tissues. Different adipogenic and lipolytic effects between subcutaneous adipose tissue (SAT) and visceral adipose tissue (VAT) are likely to play a role in GCs induced fat differential distribution. Wnt/β-catenin signaling pathway is one of the most important regulators in adipogenesis. Adipose triglyceride lipase (ATGL) and hormone sensitive lipase (HSL) are the major lipases contributing to lipolysis. In the present study, we measured fat depot masses and the expression of Wnt/β-catenin signaling pathway and lipolytic enzymes of female Sprague-Dawley rats treated with or without methylprednisolone. We assessed the roles of Wnt/β-catenin signaling pathway and lipolytic enzymes in fat differential distribution between SAT and VAT. Our data suggested that methylprednisolone could inhibit Wnt/β-catenin signaling pathway in SAT and VAT, increase the expression of ATGL and HSL in SAT, and decrease the expression of ATGL and HSL in VAT. The differential expression of lipolysis enzymes induced by methylprednisolone between SAT and VAT might play a crucial role in fat distribution. Those findings would offer novel insights into the mechanisms of GCs induced fat distribution. Copyright © 2014 Elsevier Inc. All rights reserved.

  11. Molecular characterization of a proteolysis-resistant lipase from Bacillus pumilus SG2

    Directory of Open Access Journals (Sweden)

    R. Sangeetha

    2014-06-01

    Full Text Available Proteolysis-resistant lipases can be well exploited by industrial processes which employ both lipase and protease as biocatalysts. A proteolysis resistant lipase from Bacillus pumilus SG2 was isolated, purified and characterized earlier. The lipase was resistant to native and commercial proteases. In the present work, we have characterized the lip gene which encodes the proteolysis-resistant lipase from Bacillus pumilus SG2. The parameters and structural details of lipase were analysed. The lip gene consisted of 650 bp. The experimental molecular weight of SG2 lipase was nearly double that of its theoretical molecular weight, thus suggesting the existence of the functional lipase as a covalent dimer. The proteolytic cleavage sites of the lipase would have been made inaccessible by dimerisation, thus rendering the lipase resistant to protease.

  12. IMMOBILIZATION OF LIPASE FROM PORCINE PANCREAS ON POLY (METHYL ACRYLATE)COPOLYMERS

    Institute of Scientific and Technical Information of China (English)

    XuHuixian; LiMinqin; 等

    1994-01-01

    A series of poly(methyl acrylate) copolymers of different pore structures were synthesized and functionalized by polyethylene polyamine.The lipase from porcine pancreas was adsorbed on these polymer carriers. It was found that the proe structure and functional group were basic factors which affected the activity of immobilized lipase,The optimal conditions for adsorbing lipase were studied and the effects of pH,ionic strength and temperature on the immobilized lipase were compared with those on the dissolved lipase.

  13. Culture Conditions of Psychrotrophic Fungus, Penicillium chrysogenum and Its Lipase Characteristics

    OpenAIRE

    Bsncerz, Renata; Ginalska, Grazyna; Leonowicz, Andrzej; Oga, Shoji

    2007-01-01

    Among 97 fungal strains from the soil collected from the high mountain areas in the Jeju Island, Korea, Penicillium chrysogenum 9 was found to be the best lipase producer. Its lipase productivity reached 42 U/ml in the culture medium. Factors affecting lipase production by Penicillium chrysogenum 9 were studied using fermentation media of different chemical compositions. Under optimal conditions we noted a 1.6-fold increase of lipase activity. The maximum lipase activity was 68 U/ml of cultur...

  14. Lysophosphatidylcholine synthesis by lipase-catalyzed ethanolysis.

    Science.gov (United States)

    Yang, Guolong; Yang, Ruoxi; Hu, Jingbo

    2015-01-01

    Lysophosphatidylcholine (LPC) is amphiphilic substance, and possesses excellent physiological functions. In this study, LPC was prepared through ethanolysis of phosphatidylcholine (PC) in n-hexane or solvent free media catalyzed by Novozym 435 (from Candida antarctica), Lipozyme TLIM (from Thermomcyces lanuginosus) and Lipozyme RMIM (from Rhizomucor miehei). The results showed that three immobilized lipases from Candida Antarctica, Thermomcyces lanuginosus and Rhizomucor miehei could catalyze ethanolysis of PC efficiently. In n-hexane, the LPC conversions of ethanolysis of PC catalyzed by Novozyme 435, Lipozyme TLIM and Lipozyme RMIM could reach to 98.5 ± 1.6%, 94.6 ± 1.4% and 93.7 ± 1.8%, respectively. In solvent free media, the highest LPC conversions of ethanolysis of PC catalyzed by Novozyme 435, Lipozyme TL IM and Lipozyme RM IM were 97.7 ± 1.7%, 93.5 ± 1.2% and 93.8 ± 1.9%, respectively. The catalytic efficiencies of the three lipases were in the order of Novozyme 435 > Lipozyme TLIM > Lipozyme RMIM. Furthermore, their catalytic efficiencies in n-hexane were better than those in solvent free media.

  15. Pancreatic Lipase Inhibitory Effects of Mangosteen Pericarps

    Directory of Open Access Journals (Sweden)

    Xinjie Lin

    2014-03-01

    Full Text Available Pancreatic lipase plays a key role in the digestion and absorption of lipids, inhibition of Pancreatic Lipase (PL is considered as a new approach to obesity treatment. The objective of the present study was to find PL inhibitors from natural food sources. Eighteen natural food products sampled from local supermarkets in Zhuhai were tested for PL inhibitory activity using a copper-soap photometric method. Among the samples tested, the crude extracts from mangosteen pericarp, lemon pulp, celery, cucumber and dry longan were found to be able to suppress the PL activity to different extents, while dry red chili, fresh green chili and dry clove exhibited a promotion effect on the PL. Shiitake mushroom, green bell pepper, lemon peel and spices (ginger, oregano leaf, bay leaf, cinnamon and dry tangerine showed no significant influence either on the inhibition or promotion. The crude extract of mangosteen pericarp was further fractioned to trace active fractions. It was found that the n-butanol fraction was the major contributor to the PL-inhibitory effect of mangosteen pericarp and the inhibition rate was 43.9% at the concentration of 1 mg/mL, the IC50 value was 0.918 mg/mL. Mangosteen pericarp is worthy of utilization as functional food constituents for the prevention and treatment of obesity.

  16. Influence of environmental factors on lipase production by Lactobacillus plantarum.

    Science.gov (United States)

    Lopes, M de F; Cunha, A E; Clemente, J J; Carrondo, M J; Crespo, M T

    1999-02-01

    A strain of Lactobacillus plantarum, DSMZ 12028 (Deutsch Sammlung von Mikroorganismen und Zellkulturen), isolated from a Portuguese dry fermented sausage, "chouriço", was found to produce true lipase, producing free fatty acids from triolein (olive oil). This enzymatic activity was found in whole cells, but was negligible in comparison to lipolytic activity in culture supernatant. Therefore, only extracellular activity was studied. The effect of pH, temperature and glucose concentration on extracellular lipase production was studied in continuously stirred tank reactors, the first time this technology has been used to study the production of this enzyme in lactobacilli. Maximum lipase production was achieved at a pH of 5.5 and 30 degrees C and was kept at a significant level over a wide range of dilution rates (0.05-0.4 h-1); the production of lipase was still significant for low pH values, temperature and glucose concentration, conditions that are close to the ones present during chouriço ripening. The effect of glucose concentration was also studied in a batch system. The control of lipase production was found to be related both to glucose concentration in the medium and to the growth rate/dilution rate. Glucose concentration was found to be important for fast lipase production, although it did not influence the maximum lipase activity reached in a batch culture.

  17. Lipase production in lipolytic yeast from Wonorejo mangrove area

    Science.gov (United States)

    Alami, Nur Hidayatul; Nasihah, Liziyatin; Umar, Rurin Luswidya Artaty; Kuswytasari, Nengah Dwianita; Zulaika, Enny; Shovitri, Maya

    2017-06-01

    Lipase is an enzyme that is often used in industry and become a commercial enzyme. One group of microorganisms capable of producing lipase is a yeast. This study aims to screen yeast from Wonorejo mangrove that potential to produce lipase and to optimize the production of these enzymes. Screening test include the measurement of lipolytic index and value of fatty acid. Yeast with the best value of fatty acid will be continued to the measurement of lipase activity. It is affected by several environmental factors, such as pH, temperature, and incubation time. This research was conducted to observe the optimization variation on environmental factors combination to produce lipase. Lipase activity was tested by using p-Nitrophenyl Palmitate (pNPP). Absorbency was measured by spectrofotometer on wavelength of 410 nm. Measurement of the enzyme activity was done by interpolating the absorbance values on the p-nitrophenol standard curve then calculated by the formula. All data were analyzed by using descriptive quantitative method. The results show that the highest lypolityc index was 2.08. The highest value of fatty acid was 0.49 that was reached on 168 hours of incubation. Candida W3.8 expressed the highest lypolylitic potential. The optimum environment to produce lipase by Candida W 3.8 was on 120 hours of incubation time, in temperature range of 27°C - 45°C and pH range of 4,5 - 7.

  18. The perception of gibberellins: clues from receptor structure.

    Science.gov (United States)

    Ueguchi-Tanaka, Miyako; Matsuoka, Makoto

    2010-10-01

    The discovery of GID1, a soluble receptor for gibberellins (GAs), has revealed new insights into how GA is perceived. X-ray analysis has demonstrated similarities in the tertiary structure of GID1 to hormone sensitive lipase (HSL), and the GA-binding pocket of GID1 corresponds to the active site of HSL. X-ray analysis has also revealed the structural basis of the GA-GID1 interaction, and evolutionary aspects of GID1 have been discovered by comparison to GID1 from non-flowering plants. Recent studies have also demonstrated the complexity of GA signaling in Arabidopsis, which is mediated by three GID1 and five DELLA proteins. Finally, mechanistic and structural similarities for hormone signaling are compared for GA, auxin and abscisic acid, three hormones where the receptor protein structure was recently described.

  19. Primary defects in lipolysis and insulin action in skeletal muscle cells from type 2 diabetic individuals

    DEFF Research Database (Denmark)

    Kase, E. T.; Feng, Y. Z.; Badin, P. M.

    2015-01-01

    A decrease in skeletal muscle lipolysis and hormone sensitive-lipase (HSL) expression has been linked to insulin resistance in obesity. The purpose of this study was to identify potential intrinsic defects in lipid turnover and lipolysis in myotubes established from obese and type 2 diabetic...... subjects. Lipid trafficking and lipolysis were measured by pulse-chase assay with radiolabeled substrates in myotubes from non-obese/non-diabetic (lean), obese/non-diabetic (obese) and obese/diabetic (T2D) subjects. Lipolytic protein content and level of Akt phosphorylation were measured by Western blot....... HSL was overexpressed by adenovirus-mediated gene delivery. Myotubes established from obese and T2D subjects had lower lipolysis (-30-40%) when compared to lean, using oleic acid as precursor. Similar observations were also seen for labelled glycerol. Incorporation of oleic acid into diacylglycerol...

  20. A Newly Isolated Thermostable Lipase from Bacillus sp.

    Directory of Open Access Journals (Sweden)

    Abu Bakar Salleh

    2011-05-01

    Full Text Available A thermophilic lipolytic bacterium identified as Bacillus sp. L2 via 16S rDNA was previously isolated from a hot spring in Perak, Malaysia. Bacillus sp. L2 was confirmed to be in Group 5 of bacterial classification, a phylogenically and phenotypically coherent group of thermophilic bacilli displaying very high similarity among their 16S rRNA sequences (98.5–99.2%. Polymerase chain reaction (PCR cloning of L2 lipase gene was conducted by using five different primers. Sequence analysis of the L2 lipase gene revealed an open reading frame (ORF of 1251 bp that codes for 417 amino acids. The signal peptides consist of 28 amino acids. The mature protein is made of 388 amino acid residues. Recombinant lipase was successfully overexpressed with a 178-fold increase in activity compared to crude native L2 lipase. The recombinant L2 lipase (43.2 kDa was purified to homogeneity in a single chromatography step. The purified lipase was found to be reactive at a temperature range of 55–80 °C and at a pH of 6–10. The L2 lipase had a melting temperature (Tm of 59.04 °C when analyzed by circular dichroism (CD spectroscopy studies. The optimum activity was found to be at 70 °C and pH 9. Lipase L2 was strongly inhibited by ethylenediaminetetraacetic acid (EDTA (100%, whereas phenylmethylsulfonyl fluoride (PMSF, pepstatin-A, 2-mercaptoethanol and dithiothreitol (DTT inhibited the enzyme by over 40%. The CD spectra of secondary structure analysis showed that the L2 lipase structure contained 38.6% α-helices, 2.2% ß-strands, 23.6% turns and 35.6% random conformations.

  1. Fsp27 Inhibits Lipolysis by Excluding HSL from Lipid Droplet Surface%Fsp27通过抑制HSL的脂滴定位调控脂肪水解

    Institute of Scientific and Technical Information of China (English)

    李聪; 周林康; 李蓬

    2014-01-01

    Fsp27是CIDE蛋白家族的一员,其特异性地在脂肪组织中表达并定位于脂滴表面,促进脂滴融合增大和脂肪积累.Fsp27敲除小鼠表现出胰岛素敏感性增强,有较高的能量消耗,并且可以抵抗高脂食物引起的肥胖,但Fsp27是否直接参与脂肪水解的调控过程并不清楚.本研究发现,在3T3-L1脂肪细胞中基因沉默Fsp27导致脂肪水解速率上升,并且这种上升是由激素敏感型脂肪酶(HSL)所介导.进一步在3T3-L1前脂肪细胞中过表达Fsp27以及HSL,对其定位的观察结果显示,Fsp27可以显著地抑制HSL在脂滴表面的定位.本研究表明,在脂肪组织中,Fsp27能够直接影响HSL在脂滴表面的定位,进而抑制脂肪水解速率,导致脂类积累.%Fat-specific protein 27 (Fsp27),a member of the CIDE family proteins,has been shown to be a lipid droplet-associated protein that promotes lipid storage in adipose tissue.Fsp27 knock-out mice have increased energy expenditure and are resistant to diet-induced obesity.Fsp27-induced lipid storage is mediated either by enhancing lipid droplet fusion or inhibiting lipolysis.However,the mechanism by which Fsp27 controls lipolysis remains unclear.Here,we observed that increased lipolysis in Fsp27 knockdown adipocytes was significantly abrogated when HSL was knocked down.In addition,we found that when co-expressed with Fsp27,HSL was excluded from lipid droplet surface.Furthermore,HSL exclusion was dependent on the C-terminal region of Fsp27.Our data indicate that Fsp27 regulates lipolysis by inhibiting the lipid droplet association of HSL,providing a novel mechanism for the regulation of lipid storage in adipose tissue and the development of obesity.

  2. Characterization of a highly thermostable extracellular lipase from Lactobacillus plantarum.

    Science.gov (United States)

    Lopes, Maria de Fátima Silva; Leitão, Ana Lúcia; Regalla, Manuela; Marques, J J Figueiredo; Carrondo, Manuel José Teixeira; Crespo, Maria Teresa Barreto

    2002-06-01

    After screening for the presence of lipase activity in lactobacilli isolated from "chouriço", a traditional Portuguese dry fermented sausage, a strain of Lactobacillus plantarum (DSMZ 12028) was chosen for extracellular lipase characterisation and purification. Proteinase K did not significantly affect lipolytic activity, as opposed to trypsin, which completely eliminated this activity. Among NaCl, Ca2+, EDTA, BSA, glycerol, Mn2+ and Mg2+, only Mn2+ and Mg2+ stimulated the lipase. Purification by gel filtration chromatography and gel electrophoresis revealed four bands, between 98 and 45 kDa, all with lipolytic activity against olive oil.

  3. Endothelial and lipoprotein lipases in human and mouse placenta

    DEFF Research Database (Denmark)

    Lindegaard, Marie Louise Skakkebæk; Olivecrona, Gunilla; Christoffersen, Christina;

    2005-01-01

    Placenta expresses various lipase activities. However, a detailed characterization of the involved genes and proteins is lacking. In this study, we compared the expression of endothelial lipase (EL) and LPL in human term placenta. When placental protein extracts were separated by heparin...... protein associated with both cell types. In mouse placentas, lack of LPL expression resulted in increased EL mRNA expression. These results suggest that the cellular expression of EL and LPL in human placenta is different. Nevertheless, the two lipases might have overlapping functions in the mouse...... placenta. Our data also suggest that the major portions of both proteins are stored in an inactive form in human term placenta....

  4. Stability of Surfactant—coated Candida Rugosa Lipase in Isooctane

    Institute of Scientific and Technical Information of China (English)

    宋宝东; 邢爱华; 吴金川; 王世昌

    2003-01-01

    The stability of Candida rugosa lipase coated with glutamic acid didodecyl ester ribitol amide was investigated taking esterification of lauryl alcohol and lauric acid in isooctane as a model reaction.At 30℃,the half-life of the activity of the coated lipase was ca 10h,the enzyme activity became less changed after 12h and the residual activity was 39% of the initial value ,The coated lipase obeyed a first-order deactivation model with a deactivation energy of 29.9 J.mol-1.

  5. Obtaining lipases from byproducts of orange juice processing.

    Science.gov (United States)

    Okino-Delgado, Clarissa Hamaio; Fleuri, Luciana Francisco

    2014-11-15

    The presence of lipases was observed in three byproducts of orange juice processing: peel, core and frit. The enzymes were characterised biochemically over a wide pH range from neutral (6-7) to alkaline (8-9). The optimal temperature for the activity of these byproducts showed wide range at 20°C to 70°C, indicating fairly high thermostability. The activities were monitored on p-NP-butyrate, p-NP-laurate and p-NP-palmitate. For the first time, lipase activity was detected in these residues, reaching 68.5 lipase U/g for the crude extract from fractions called frit.

  6. Maximization of Intracellular Lipase Production in a Lipase-Overproducing Mutant Derivative of Rhizopus oligosporus DGM 31: A Kinetic Study

    Directory of Open Access Journals (Sweden)

    Tehreema Iftikhar

    2008-01-01

    Full Text Available Regulation and maximization of lipase production in a mutant derivative of R. oligosporus has been investigated using different substrates, inoculum sizes, pH of the medium, temperature, and nitrogen sources in shake flask experiments and batch fermentation in a fermentor. The production of intracellular lipase was improved 3 times following medium optimization involving one-at-a-time approach and aeration in the fermentor. Interestingly, intracellular lipase was poorly induced by oils, instead its production was induced by sugars, mainly starch, lactose, sucrose, xylose, glucose and glycerol. Dependent variables studied were cell mass, lipase activity, lipase yield, lipase specific and volumetric rate of formation. It was confirmed that lipase production in the derepressed mutant is sufficiently uncoupled from catabolite repression. The results of average specific productivities at various temperatures worked out according to the Arrhenius equation revealed that mutation decreased the magnitude of enthalpy and entropy demand in the inactivation equilibrium during product formation, suggesting that mutation made the metabolic network of the organism thermally more stable. The highest magnitudes of volumetric productivity (QP=490 IU/(L·h and other product attributes of lipase formation occurring on optimized medium in the fermentor are greater than the values reported by other workers. The purified enzyme is monomeric in nature and exhibits stability up to 80 °C and pH=6.0–8.0. Activation energy, enthalpy and entropy of catalysis at 50 °C, and magnitudes of Gibbs free energy for substrate binding, transition state stabilization and melting point indicated that this lipase is highly thermostable.

  7. In vitro lipolysis tests on lipid nanoparticles: comparison between lipase/co-lipase and pancreatic extract.

    Science.gov (United States)

    Jannin, Vincent; Dellera, Eleonora; Chevrier, Stéphanie; Chavant, Yann; Voutsinas, Christophe; Bonferoni, Cristina; Demarne, Frédéric

    2015-01-01

    Solid lipid nanoparticles (SLNs) and nanostructured lipid carriers (NLC) are lipid nanocarriers aimed to the delivery of drugs characterized by a low bioavailability, such as poorly water-soluble drugs and peptides or proteins. The oral administration of these lipid nanocarriers implies the study of their lipolysis in presence of enzymes that are commonly involved in dietary lipid digestion in the gastrointestinal tract. In this study, a comparison between two methods was performed: on one hand, the lipase/co-lipase assay, commonly described in the literature to study the digestion of lipid nanocarriers, and on the other hand, the lipolysis test using porcine pancreatic extract and the pH-stat apparatus. This pancreatic extract contains both the pancreatic lipase and carboxyl ester hydrolase (CEH) that permit to mimic in a biorelevant manner the duodenal digestive lipolysis. The test was performed by means of a pH-stat apparatus to work at constant pH, 5.5 or 6.25, representing respectively the fasted or fed state pH conditions. The evolution of all acylglycerol entities was monitored during the digestion by sampling the reaction vessel at different time points, until 60 min, and the lipid composition of the digest was analyzed by gas chromatography. SLN and NLC systems obtained with long-chain saturated acylglycerols were rapidly and completely digested by pancreatic enzymes. The pH-stat titration method appears to be a powerful technique to follow the digestibility of these solid lipid-based nanoparticles.

  8. Primary defects in lipolysis and insulin action in skeletal muscle cells from type 2 diabetic individuals.

    Science.gov (United States)

    Kase, Eili T; Feng, Yuan Z; Badin, Pierre-Marie; Bakke, Siril S; Laurens, Claire; Coue, Marine; Langin, Dominique; Gaster, Michael; Thoresen, G Hege; Rustan, Arild C; Moro, Cedric

    2015-09-01

    A decrease in skeletal muscle lipolysis and hormone sensitive-lipase (HSL) expression has been linked to insulin resistance in obesity. The purpose of this study was to identify potential intrinsic defects in lipid turnover and lipolysis in myotubes established from obese and type 2 diabetic subjects. Lipid trafficking and lipolysis were measured by pulse-chase assay with radiolabeled substrates in myotubes from non-obese/non-diabetic (lean), obese/non-diabetic (obese) and obese/diabetic (T2D) subjects. Lipolytic protein content and level of Akt phosphorylation were measured by Western blot. HSL was overexpressed by adenovirus-mediated gene delivery. Myotubes established from obese and T2D subjects had lower lipolysis (-30-40%) when compared to lean, using oleic acid as precursor. Similar observations were also seen for labelled glycerol. Incorporation of oleic acid into diacylglycerol (DAG) and free fatty acid (FFA) level was lower in T2D myotubes, and acetate incorporation into FFA and complex lipids was also lower in obese and/or T2D subjects. Both protein expression of HSL (but not ATGL) and changes in DAG during lipolysis were markedly lower in cells from obese and T2D when compared to lean subjects. Insulin-stimulated glycogen synthesis (-60%) and Akt phosphorylation (-90%) were lower in myotubes from T2D, however, overexpression of HSL in T2D myotubes did not rescue the diabetic phenotype. In conclusion, intrinsic defects in lipolysis and HSL expression co-exist with reduced insulin action in myotubes from obese T2D subjects. Despite reductions in intramyocellular lipolysis and HSL expression, overexpression of HSL did not rescue defects in insulin action in skeletal myotubes from obese T2D subjects.

  9. Clofazimine modulates the expression of lipid metabolism proteins in Mycobacterium leprae-infected macrophages.

    Directory of Open Access Journals (Sweden)

    Yang Degang

    Full Text Available Mycobacterium leprae (M. leprae lives and replicates within macrophages in a foamy, lipid-laden phagosome. The lipids provide essential nutrition for the mycobacteria, and M. leprae infection modulates expression of important host proteins related to lipid metabolism. Thus, M. leprae infection increases the expression of adipophilin/adipose differentiation-related protein (ADRP and decreases hormone-sensitive lipase (HSL, facilitating the accumulation and maintenance of lipid-rich environments suitable for the intracellular survival of M. leprae. HSL levels are not detectable in skin smear specimens taken from leprosy patients, but re-appear shortly after multidrug therapy (MDT. This study examined the effect of MDT components on host lipid metabolism in vitro, and the outcome of rifampicin, dapsone and clofazimine treatment on ADRP and HSL expression in THP-1 cells. Clofazimine attenuated the mRNA and protein levels of ADRP in M. leprae-infected cells, while those of HSL were increased. Rifampicin and dapsone did not show any significant effects on ADRP and HSL expression levels. A transient increase of interferon (IFN-β and IFN-γ mRNA was also observed in cells infected with M. leprae and treated with clofazimine. Lipid droplets accumulated by M. leprae-infection were significantly decreased 48 h after clofazimine treatment. Such effects were not evident in cells without M. leprae infection. In clinical samples, ADRP expression was decreased and HSL expression was increased after treatment. These results suggest that clofazimine modulates lipid metabolism in M. leprae-infected macrophages by modulating the expression of ADRP and HSL. It also induces IFN production in M. leprae-infected cells. The resultant decrease in lipid accumulation, increase in lipolysis, and activation of innate immunity may be some of the key actions of clofazimine.

  10. A lipase with broad temperature range from an alkaliphilic gamma-proteobacterium isolated in Greenland

    DEFF Research Database (Denmark)

    Schmidt, Mariane; Larsen, Dorte Møller; Stougaard, Peter

    2010-01-01

    A gamma-proteobacterium related to the genera Alteromonadales and Pseudomonadales , isolated from a cold and alkaline environment in Greenland, has been shown to produce a lipase active between 5 ° C and 80 ° C, with optimal activity at 55 ° C and pH 8. PCR-based screening of genomic DNA from...... the isolated bacterium, followed by genome walking, resulted in two complete open reading frames, which were predicted to encode a lipase and its helper protein, a lipase foldase. The amino acid sequence derived for the lipase showed resemblance to lipases from Pseudomonas , Rhodoferax, Aeromonas and Vibrio...... . The two genes were cloned into different expression systems in E. coli with or without a putative secretion sequence, but despite the fact that both recombinant lipase and lipase foldase were observed on SDS–PAGE, no recombinant lipase activity was detected. Attempts to refold the recombinant lipase...

  11. Stability of immobilized candida sp. 99-125 Lipase for biodiesel production

    Energy Technology Data Exchange (ETDEWEB)

    Lu, J. [Beijing Bioprocess Key Laboratory, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing (China); Bioengineering Department, Zhengzhou University, Zhengzhou (China); Deng, L.; Nie, K.; Wang, F.; Tan, T. [Beijing Bioprocess Key Laboratory, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing (China)

    2012-12-15

    The stability of the immobilized lipase from Candida sp. 99-125 during biodiesel production was investigated. The lipase was separately incubated in the presence of various reaction components such as soybean oil, oleic acid methyl ester, n-hexane, water, methanol, and glycerol, or the lipase was stored at 60, 80, 100 and 120 C. Thereafter the residual lipase activity was determined by methanolysis reaction. The results showed that the lipase was rather stable in the reaction media, except for methanol and glycerol. The stability study performed in a reciprocal shaker indicated that enzyme desorption from the immobilized lipase mainly contributed to the lipase inactivation in the water system. So the methanol and glycerol contents should be controlled more precisely to avoid lipase inactivation, and the immobilization method should be improved with regard to lipase desorption. (Copyright copyright 2012 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

  12. Selection and optimization of extracellular lipase production using ...

    African Journals Online (AJOL)

    Pedro

    2014-01-22

    Jan 22, 2014 ... in their catalytic properties, and can be obtained by solid- state fermentation or by submerged fermentation. (Annibale et al., 2006; Rigo et al., 2010; ..... A microwave-assisted micro- assay for lipases. Anal. Bioanal. Chem.

  13. Recent developments in lipase-catalyzed synthesis of polyesters.

    Science.gov (United States)

    Kobayashi, Shiro

    2009-02-18

    Polyester synthesis by lipase catalyst involves two major polymerization modes: i) ring-opening polymerization of lactones, and, ii) polycondensation. Ring-opening polymerization includes the finding of lipase catalyst; scope of reactions; polymerization mechanism; ring-opening polymerization reactivity of lactones; enantio-, chemo- and regio-selective polymerizations; and, chemoenzymatic polymerizations. Polycondensation includes polymerizations involving condensation reactions between carboxylic acid and alcohol functional groups to form an ester bond. In most cases, a carboxylic acid group is activated as an ester form, such as a vinyl ester. Many recently developed polymerizations demonstrate lipase catalysis specific to enzymatic polymerization and appear very useful. Also, since lipase-catalyzed polyester synthesis provides a good opportunity for conducting "green polymer chemistry", the importance of this is described.

  14. Lipase-producing fungi for potential wastewater treatment and ...

    African Journals Online (AJOL)

    Lipase-producing fungi for potential wastewater treatment and bioenergy production. ... as well as for the production of biodiesel from oil and residual grease, due to its greater stability, possibility of reuse, and lower cost. ... Article Metrics.

  15. Isolation, purification and properties of lipase from Pseudomonas ...

    African Journals Online (AJOL)

    Isolation, purification and properties of lipase from Pseudomonas aeruginosa. ... medium agar containing Tween 80 or olive oil as the only source of carbon. ... through the purification procedure of ammonium sulphate precipitation and diethyl ...

  16. Lipase Activity among Bacteria Isolated from Amazonian Soils

    Directory of Open Access Journals (Sweden)

    André Luis Willerding

    2011-01-01

    Full Text Available The objective of this study was to select lipase-producing bacteria collected from different counties of the Amazon region. Of the 440 bacteria strains, 181 were selected for the lipase assay in qualitative tests at Petri dishes, being 75 (41% lipase positive. The enzymatic index was determined during fifteen days at different temperatures (30°, 35°, 40°, and 45°C. The highest lipase activity was observed within 72 hours at 30°C. Twelve bacteria strains presented an index equal to or greater than the standard used like reference, demonstrating the potential of microbial resource. After the bioassay in Petri dishes, the selected bacteria strains were analyzed in quantitative tests on p-nitrophenyl palmitate (p-NPP. A group of the strains was selected for other phases of study with the use in oleaginous substrates of the Amazonian flora, aiming for the application in processes like oil biotransformation.

  17. Lipase catalyzed synthesis of epoxy-fatty acids

    Institute of Scientific and Technical Information of China (English)

    CHEN, Qian; LI, Zu-Yi

    2000-01-01

    Lipase catalyzed synthesis of epoxy-fatty acidas from unsaturated carboxylic acids was investigated.Under mild conditions unsaturated arboxylic acids were convcveed to peroxide,then the unsaturated peroxycarboxylic acids epoxidised the C=C bond of themselves

  18. Isolation and characterization of lipase-producing Bacillus strains ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-08-04

    spilled areas of the coconut oil extracting industry in a sterile container for the ... By using different substrates sources such as olive oil, coconut oil and sunflower oil, their ..... characterization and application of lipases. Biotechnol.

  19. Production of extracellular lipase by a new strain Staphylococcus ...

    African Journals Online (AJOL)

    SAM

    2014-07-09

    Jul 9, 2014 ... detrimental effect on microorganisms and so they lose their activity and .... +. Oxidase. -. Urease. -. Nitrate Reduction Test. +. Glucose Fermentation. + ... on lipase production which might be due to the inhibition by fatty acid ...

  20. Lipase Activity in Fermented Oil Seeds of Africa Locust Bean ...

    African Journals Online (AJOL)

    acer

    Castor Seeds (Ricinu Communis) and African Oil Bean (Pentaclethra Macrophylla). A.A. Liman*, P. ... The peak lipase activity for fermented Africa locust bean, Castor seed, and African oil bean were ..... fermented vegetable proteins. World.

  1. KINETICS OF HYDROLYSIS OF TRIBUTYRIN BY LIPASE

    Directory of Open Access Journals (Sweden)

    SULAIMAN AL-ZUHAIR

    2006-06-01

    Full Text Available Kinetics of the enzymatic hydrolysis of tributyrin using lipase has been investigated. The initial rate of reaction was determined experimentally at different substrate concentration by measuring the rate of butyric acid produced. Michaels-Menten kinetic model has been proposed to predict the initial rate of hydrolysis of tributyrin in micro-emulsion system. The kinetic parameters were estimated by fitting the data to the model using three methods, namely, the Lineweaver-Burk, Edie-Hofstee and Hanes methods. The Michaels-Menten model with the constant predicted by Edie-Hofstee and Hanes methods predicted the initial rate of reaction at various substrate concentrations better than the model with the constant predicted Lineweaver-Burk method, especially at high substrate concentrations.

  2. Low-Cost Biodegradation and Detoxification of Textile Azo Dye C.I. Reactive Blue 172 by Providencia rettgeri Strain HSL1

    Directory of Open Access Journals (Sweden)

    Harshad Lade

    2015-01-01

    Full Text Available Present study focuses on exploitation of agricultural waste wheat bran (WB as growth medium for degradation of textile azo dye C.I. Reactive Blue 172 (RB 172 using a single bacterium P. rettgeri strain HSL1 (GenBank accession number JX853768.1. The bacterium was found to completely decolorize 50 mg L−1 of dye RB 172 within 20 h at 30 ± 0.2°C under microaerophilic incubation conditions. Additionally, significant reduction in COD (85% and TOC (52% contents of dye decolorized medium was observed which suggested its mineralization. Induction in the activities of azoreductase (159% and NADH-DCIP reductase (88% provided an evidence for reductive cleavage of dye RB 172. The HPLC, FTIR, and GC-MS analysis of decolorized products confirmed the degradation of dye into various metabolites. The proposed metabolic pathway for biodegradation of RB 172 has been elucidated which showed the formation of 2 intermediate metabolites, namely, 4-(ethenylsulfonyl aniline and 1-amino-1-(4-aminophenyl propan-2-one. The acute and phytotoxicity evaluation of degraded metabolites suggests that bacterial strain favors the detoxification of dye RB 172. Thus, WB could be utilized as a low-cost growth medium for the enrichment of bacteria and their further use for biodegradation of azo dyes and its derivatives containing wastes into nontoxic form.

  3. LIPASE-CATALYZED TRANSESTERIFICATION OF PALM KERNEL OIL WITH DIALKYLCARBONATES

    OpenAIRE

    Tjahjono Herawan; M. Rüsch Gen. Klaas

    2014-01-01

    Lipase-catalyzed transesterifications-especially in a solvent-free medium-are important for industrial applications because such systems would have an enormous advantage by avoiding the problem of separation, toxicity and flammability of organic solvents. However, the organic solvent-free alcoholysis, especially methanolysis, does not give high conversions. The same problem also occurs when ethyl or methyl acetate are used as acyl acceptors. The main problems of lipase-catalyzed organic solve...

  4. Anaerobic biodegradability of dairy wastewater pretreated with porcine pancreas lipase

    OpenAIRE

    2010-01-01

    Lipids-rich wastewater was partial hydrolyzed with porcine pancreas lipase and the efficiency of the enzymatic pretreatment was verified by the comparative biodegradability tests (crude and treated wastewater). Alternatively, simultaneous run was carried out in which hydrolysis and digestion was performed in the same reactor. Wastewater from dairy industries and low cost lipase preparation at two concentrations (0.05 and 0.5% w.v-1) were used. All the samples pretreated with enzyme showed a p...

  5. Investigation of Lipase Production by Milk Isolate Serratia rubidaea

    Directory of Open Access Journals (Sweden)

    Palanichamy Esakkiraj

    2008-01-01

    Full Text Available Production of extracellular lipase in submerged culture of Serratia rubidaea has been investigated. The lipase production was optimized in shake flask experiments. The observed pH and temperature range optimum for maximum lipase production were 7–8 and 30–40 °C, respectively. With a selected nitrogen source, casein ((6.5±0.015 U/mL and soytone ((9.4±0.02 U/mL were suitable substrates for accelerating lipase production. The optimized concentration of casein and soytone was 24 g/L ((9.95±0.02 U/mL and 5 g/L ((14.8±0.03 U/mL, respectively. The effect of carbon source on lipase production indicated that starch was suitable substrate to maximize lipase production ((15.60±0.20 U/mL and the optimum concentration registered was 4 g/L ((17.46±0.20 U/mL. Investigating the effect of lipids and surfactants showed that the gingily oil ((20.52±0.20 U/mL and Tween 20 ((27.10±0.01 U/mL were suitable substrates for maximizing lipase production, and the optimum concentrations registered were 15 mL/L ((23.15±0.24 U/mL and 6 mL/L ((34.20±0.01 U/mL, respectively. Partial purification of lipase indicated that the molecular mass of partially purified enzyme was 54 kDa.

  6. 激素敏感脂肪酶(HSL)及其与代谢性疾病关系的研究

    Institute of Scientific and Technical Information of China (English)

    韩丽红; 李学英; 王大忠

    2005-01-01

    激素敏感脂肪酶(hormone-sensitive lipase,HSL)是动物脂肪分解代谢的限速酶。它通过催化水解储存在脂肪组织中的甘油三酯释放出游离的脂肪酸,以满足机体的能量需要。HSL主要在动物的白色脂肪组织中表达,同时也在肝、骨骼肌、睾丸等组织中有少量的表达。它的活性受体内多种激素调控。在机体正常的生理条件下,HSL发挥着较多的功能,如影响脂肪的代谢、类固醇的合成、精子的生成、胰岛素的分泌等。HSL表达异常或HSL基因(hsl)多态性与人类代谢性疾病如肥胖、胰岛素抵抗、二型糖尿病和高脂血症等之间关系的研究目前已经取得一些进展,现综述如下。

  7. Les lipases immobilisées et leurs applications

    Directory of Open Access Journals (Sweden)

    Thonart P.

    2008-01-01

    Full Text Available Immobilized lipases and their applications. Lipases are able to catalyse the hydrolysis of glyceridic esters in aqueous media and the synthesis of esters in non-aqueous media. They are thus able to catalyse numerous reactions of industrial interest. Whether it is by inclusion, by adsorption or by covalent link, the immobilisation of lipases aims at conferring them a good stability that enables a reuse of the enzymes after a reaction and the development of continuous processes. The reactions of triglycerides hydrolysis constitute main applications for immobilised lipases, however their use in different types of esterification reactions has also arose: there exist processes involving reactions of transesterification, of interesterification or of esters synthesis. The production of structured lipids by interesterification is one example. Although the reaction conditions dissent from those of hydrolysis, the same lipases have been used in both cases. A lipase specifically adapted for esterification though would be a highly capable tool: a series of strategies is in progress in order to reach this goal.

  8. Enzymatic production of biodiesel from canola oil using immobilized lipase

    Energy Technology Data Exchange (ETDEWEB)

    Dizge, Nadir; Keskinler, Buelent [Department of Environmental Engineering, Gebze Institute of Technology, Gebze 41400 (Turkey)

    2008-12-15

    In the present work, a novel method for immobilization of lipase within hydrophilic polyurethane foams using polyglutaraldehyde was developed for the immobilization of Thermomyces lanuginosus lipase to produce biodiesel with canola oil and methanol. The enzyme optimum conditions were not affected by immobilization and the optimum pH for free and immobilized enzyme were 6, resulting in 80% immobilization yield. Using the immobilized lipase T. lanuginosus, the effects of enzyme loading, oil/alcohol molar ratio, water concentration, and temperature in the transesterification reaction were investigated. The optimal conditions for processing 20 g of refined canola oil were: 430 {mu}g lipase, 1:6 oil/methanol molar ratio, 0.1 g water and 40 C for the reactions with methanol. Maximum methyl esters yield was 90% of which enzymatic activity remained after 10 batches, when tert-butanol was adopted to remove by-product glycerol during repeated use of the lipase. The immobilized lipase proved to be stable and lost little activity when was subjected to repeated uses. (author)

  9. Lipases at interfaces: unique interfacial properties as globular proteins.

    Science.gov (United States)

    Reis, P; Miller, R; Krägel, J; Leser, M; Fainerman, V B; Watzke, H; Holmberg, K

    2008-06-01

    The adsorption behavior of two globular proteins, lipase from Rhizomucor miehei and beta-lactoglobulin, at inert oil/water and air/water interfaces was studied by the pendant drop technique. The kinetics and adsorption isotherms were interpreted for both proteins in different environments. It was found that the adopted mathematical models well describe the adsorption behavior of the proteins at the studied interfaces. One of the main findings is that unique interfacial properties were observed for lipase as compared to the reference beta-lactoglobulin. A folded drop with a "skinlike" film was formed for the two proteins after aging followed by compression. This behavior is normally associated with protein unfolding and covalent cross-linking at the interface. Despite this, the lipase activity was not suppressed. By highlighting the unique interfacial properties of lipases, we believe that the presented work contributes to a better understanding of lipase interfacial activation and the mechanisms regulating lipolysis. The results indicate that the understanding of the physical properties of lipases can lead to novel approaches to regulate their activity.

  10. The modulation of pancreatic lipase activity by alginates.

    Science.gov (United States)

    Wilcox, Matthew D; Brownlee, Iain A; Richardson, J Craig; Dettmar, Peter W; Pearson, Jeffrey P

    2014-03-01

    Alginates are comprised of mannuronic (M) and guluronic acid (G) and have been shown to inhibit enzyme activity. Pancreatic lipase is important in dietary triacylglycerol breakdown; reducing pancreatic lipase activity would reduce triacylglycerol breakdown resulting in lower amounts being absorbed by the body. Lipase activity in the presence of biopolymers was assessed by enzymatic assay using natural and synthetic substrates. Alginate inhibited pancreatic lipase by a maximum of 72.2% (±4.1) with synthetic substrate (DGGR) and 58.0% (±9.7) with natural substrate. High-G alginates from Laminaria hyperborea seaweed inhibited pancreatic lipase to a significantly higher degree than High-M alginates from Lessonia nigrescens, showing that inhibition was related to alginate structure. High-G alginates are effective inhibitors of pancreatic lipase and are used in the food industry at low levels. They could be included at higher levels in foods without altering organoleptic qualities, potentially reduce the uptake of dietary triacylglycerol aiding in weight management.

  11. Karakterisasi ekstrak kasar lipase Rhizopus stolonifer UICC 137

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    Sri Sumiarsih

    2001-12-01

    Full Text Available There is an increasing commercial interest in enzymatic production of biologically active component, because there are a number of well-known advantages compared to chemical synthesis. One of the most valuable synthetic features of enzyme is their ability to discriminate between enantiomers of racemic substrates. Lipase have become of great interest to the chemical industries wing their usefulness in both hydrolytic and synthesis reactions. The aim of this work was to study the production of lipase by Rhizopus stolonifer UICC 137, and determine the crude lipase preparation characteristics. The lipolytic activity was determined by titrimetric method toward oil-arabic gum emultion as a substrate. The strain produced lipase at appreciable lipolytic when cultivated for 72 hours in medium containing 3% glucose and 1% olive oil. Our data suggest that the strain produced lipase since the exponential phase of its growth. Lipase with optimum lipolytic activity was obtained at late stationary phase. The optimum condition for lipolytic activity measurement were pH of 7.5 and temperature 37oC, the crude enzyme had a specific activity 20.2 unit/ mg protein, the Vmax was 15.1 mol/ min and KM was 12.5 mg/ ml. The crude enzyme retained 79.9%, 68.0% and 52.6% of its lipolytic activity, when incubated for 90 minutes at temperature of 40, 50, and 60oC respectively.

  12. OPTIMASI ISOLASI LIPASE INDIGENOUS BIJI KAKAO (Theobroma cacao L. The Optimizing of Isolation of Cocoa Bean Indogenous Lipase (Theobroma cacao L.

    Directory of Open Access Journals (Sweden)

    I D. G. Mayun Permana

    2012-05-01

    Full Text Available The aim of the research is to optimize the isolation method of cocoa bean lipase. The research is held by determining the position of lipase on cocoa bean, varying extraction medium and isolation process. The result shows that the lipase of cocoa bean is   cytosolic enzyme. The defatting process do not increase the lipase activity. Polyphenols inhibit the lipase activity, so that removal of the polyphenol will increase the activity. Blocking the polyphenol with polyvinilpolypirrolidone (PVPP will also increase the activity.The optimum consentration of PVPP is 8 %. The lipase activity will reach the highest when homogenized for 10 menit at 10,000 rpm. The best medium extraction for lipase isolation is 0.15 M phosphate buffer pH 7.5 containing sucrose 0.6 M and CaCl  1.0 mM.   ABSTRAK Penelitian ini bertujuan untuk mengoptimasi isolasi lipase indigenous biji kakao. Optimasi diawali dengan menentukan keberadaan lipase kemudian optimasi medium ekstraksi dan proses ekstraksi. Hasil penelitian menunjukkan bahwa lipase berada dalam sitosol. Penghilangan lemak tidak meningkatkan aktivitas lipase. Senyawa polifenol menghambat aktivitas lipase dan penghilangan polifenol dapat meningkatkan aktivitas lipase. Polyvinilpolypirrolidone (PVPP dapat menghambat polifenol sehingga dapat meningkatkan aktivitas lipase. Konsentrasi PVPP optimum adalah 8 % dari berat biji kakao. Proses homogenisasi optimum diperoleh dalam waktu 10 menit pada kecepatan 10.000 rpm. Medium ekstraksi untuk isolasi lipase biji kakao terbaik adalah bufer fosfat 0,15 M  dan pH 7,5 yang mengandung sukrosa 0,6 M dan 1,0 mM CaCl .

  13. Silk-Cocoon Matrix Immobilized Lipase Catalyzed Transesterification of Sunflower Oil for Production of Biodiesel

    OpenAIRE

    Sushovan Chatterjee; Dipti Yadav; Lepakshi Barbora; Pinakeswar Mahanta; Pranab Goswami

    2014-01-01

    Biodiesel from sunflower oil using lipase chemically immobilized on silk-cocoon matrix in a packed-bed bioreactor was investigated. The immobilization was demonstrated by field-emission scanning electron microscopy and activity study. The lipase loading was 738.74 U (~0.01 g lipase powder)/g-lipase-immobilized matrix. The Km (Michaelis-Menten constant) of the free and the immobilized lipase was 451.26 μM and 257.26 μM, respectively. Low Km value of the immobilized lipase is attributed to the ...

  14. Screening of lipase inhibitors from Scutellaria baicalensis extract using lipase immobilized on magnetic nanoparticles and study on the inhibitory mechanism.

    Science.gov (United States)

    Wan, Li-Hong; Jiang, Xiao-Lan; Liu, Yi-Ming; Hu, Jin-Jie; Liang, Jian; Liao, Xun

    2016-03-01

    Scutellaria baicalensis is a traditional Chinese medicinal plant possessing a wide variety of biological activities. In this work, lipase immobilized on magnetic nanoparticles (LMNPs) was used as solid phase extract absorbent for screening of lipase inhibitors from this plant. Three flavonoids were found to bind to LMNPs and were identified as baicalin, wogonin, and oroxylin A by liquid chromatography-mass spectrometry (HPLC-MS). Their IC50 values were determined to be 229.22 ± 12.67, 153.71 ± 9.21, and 56.07 ± 4.90 μM, respectively. Fluorescence spectroscopy and molecular docking were used to probe the interactions between these flavonoids and lipase. All the flavonoids quenched the fluorescence of lipase statically by forming new complexes, implying their affinities with the enzyme. The thermodynamic analysis suggested that van der Waals force and hydrogen bond were the main forces between wogonin and lipase, while hydrophobic force was the main force for the other two flavonoids. The results from a molecular docking study further revealed that all of them could insert into the pocket of lipase binding to a couple of amino acid residues.

  15. Potencial de biocatálise enantiosseletiva de lipases microbianas Potential of enantioselective biocatalysis by microbial lipases

    Directory of Open Access Journals (Sweden)

    Patrícia de O. Carvalho

    2005-08-01

    Full Text Available Microbial lipases have a great potential for commercial applications due to their stability, selectivity and broad substrate specificity because many non-natural acids, alcohols or amines can be used as the substrate. Three microbial lipases isolated from Brazilian soil samples (Aspergillus niger; Geotrichum candidum; Penicillium solitum were compared in terms of their stability and as biocatalysts in the enantioselective esterification using racemic substrates in organic medium. The lipase from Aspergillus niger showed the highest activity (18.2 U/mL and was highly thermostable, retaining 90% and 60% activity at 50 ºC and 60 ºC after 1 hour, respectively. In organic medium, this lipase provided the best results in terms of enantiomeric excess of the (S-active acid (ee = 6.1% and conversion value (c = 20% in the esterification of (R,S-ibuprofen with 1-propanol in isooctane. The esterification reaction of the racemic mixture of (R,S-2-octanol with decanoic acid proceeded with high enantioselectivity when lipase from Aspergillus niger (E = 13.2 and commercial lipase from Candida antarctica (E = 20 were employed.

  16. Simulação de bloqueios periféricos guiados por ultra-som: curva de aprendizado dos residentes de anestesiologia do CET-SMA/HSL Simulacro de bloqueos periféricos guiados por ultrasonido: curva de aprendizaje de los residentes de anestesiología del CET-SMA/HSL Simulation of ultrasound-guided peripheral nerve block: learning curve of CET-SMA/HSL anesthesiology residents

    Directory of Open Access Journals (Sweden)

    Marilia Bonifácio Baranauskas

    2008-04-01

    Full Text Available JUSTIFICATIVA E OBJETIVOS: A técnica de ultra-som tem sido cada vez mais utilizada para a realização de bloqueios de nervos periféricos. Existem poucos relatos na literatura que analisam a curva de aprendizado da técnica de ultra-som. O objetivo do estudo foi avaliar a curva de aprendizado dos residentes de Anestesiologia do CET-SMA/HSL em bloqueios periféricos guiados por ultra-som por meio de modelo experimental de gelatina. MÉTODO: Foi desenvolvido modelo experimental com cuba preenchida de gelatina e azeitona submersa. Nove residentes foram distribuídos aleatoriamente em três grupos (G1, G2 e G3 compostos, cada um, de um R1, um R2 e um R3. Os três grupos receberam explanação teórica. O G1 recebeu duas horas de treinamento prático, o G2 uma hora e o G3 não treinou. Na seqüência, os participantes foram solicitados a posicionar a agulha no ponto médio da parede da azeitona, próximo ao transdutor e reposicionar a agulha entre a azeitona e o fundo da cuba, simulando a injeção perineural do anestésico. Foram avaliadas a velocidade e eficácia das tarefas, além das falhas técnicas. RESULTADOS: O G1 apresentou média de tempo para realização das tarefas de 37,63 segundos, sem falhas técnicas; no G2 observou-se média de 64,40 segundos, ocorrendo duas falhas técnicas e o G3 apresentou média de 93,83 segundos, com 12 falhas técnicas. CONCLUSÕES: O estudo permite concluir que o maior tempo de treinamento em modelo experimental de bloqueios periféricos guiados por ultra-som melhorou a curva de aprendizado na simulação da técnica.JUSTIFICATIVA Y OBJETIVOS: La técnica de ultrasonido ha sido cada vez más utilizada para la realización de bloqueos de nervios periféricos. Existen pocos relatos en la literatura que analizan la curva de aprendizaje de la técnica de ultrasonido. El objetivo del estudio fue evaluar la curva de aprendizaje de los residentes de Anestesiología del CET-SMA/HSL en bloqueos periféricos guiados

  17. Dissecting adipose tissue lipolysis: molecular regulation and implications for metabolic disease.

    Science.gov (United States)

    Nielsen, Thomas Svava; Jessen, Niels; Jørgensen, Jens Otto L; Møller, Niels; Lund, Sten

    2014-06-01

    Lipolysis is the process by which triglycerides (TGs) are hydrolyzed to free fatty acids (FFAs) and glycerol. In adipocytes, this is achieved by sequential action of adipose TG lipase (ATGL), hormone-sensitive lipase (HSL), and monoglyceride lipase. The activity in the lipolytic pathway is tightly regulated by hormonal and nutritional factors. Under conditions of negative energy balance such as fasting and exercise, stimulation of lipolysis results in a profound increase in FFA release from adipose tissue (AT). This response is crucial in order to provide the organism with a sufficient supply of substrate for oxidative metabolism. However, failure to efficiently suppress lipolysis when FFA demands are low can have serious metabolic consequences and is believed to be a key mechanism in the development of type 2 diabetes in obesity. As the discovery of ATGL in 2004, substantial progress has been made in the delineation of the remarkable complexity of the regulatory network controlling adipocyte lipolysis. Notably, regulatory mechanisms have been identified on multiple levels of the lipolytic pathway, including gene transcription and translation, post-translational modifications, intracellular localization, protein-protein interactions, and protein stability/degradation. Here, we provide an overview of the recent advances in the field of AT lipolysis with particular focus on the molecular regulation of the two main lipases, ATGL and HSL, and the intracellular and extracellular signals affecting their activity.

  18. Study the effect of F17S mutation on the chimeric Bacillus thermocatenulatus lipase

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    Seyed Hossein Khaleghinejad

    2016-06-01

    Full Text Available Lipases (triacylglycerol acylhydrolase, EC 3.1.1.3 are one of the highest value commercial enzymes as they have potential applications in biotechnology for detergents, food, pharmaceuticals, leather, textiles, cosmetics, and paper industries; and are currently receiving considerable attention because of their potential applications in biotechnology. Bacillus thermocatenulatus Lipase 2 (BTL2 is one of the most important research targets, because of its potential industrial applications. In this study, the effect of substitution Phe17 with Ser in mutated BTL2 lipase, which conserved pentapeptide (112Ala-His-Ser-Gln-Gly116 was replaced with similar sequences (207Gly-Glu-Ser-Ala-Gly211 of Candida rugosa lipase (CLR at the nucleophilic elbow region. Docking results confirmed the mutated lipase to be better than the chimeric lipase. So, cloning was conducted, and the mutated and chimeric btl2 genes were expressed in Escherichia coli, and then the enzymes were purified by anion exchange chromatography. The mutation increased lipase lipolytic activity against most of the applied substrates, with the exception of tributyrin when compared with chimeric lipase. Further, the mutated lipase exhibited higher activity than the chimeric lipase at all temperatures. Optimum pH of the mutated lipase was obtained at pH 9.5, which was more than the chimeric one. Enzyme activity of the mutated lipase in the presence of organic solvents, detergents, and metal ions was also improved than the chimeric lipase.

  19. Attachment of Lipase on Amino Functionalized Titania Submicrospheres via Covalent Binding

    Institute of Scientific and Technical Information of China (English)

    WU Hong; LIANG Yan-peng; SHI Jia-fu; WANG Xiao-li

    2013-01-01

    A facile and effective method for immobilized lipase was presented.The titania submicrospheres were synthesized via a modified sol-gel method followed by amino functionalization through the chelation between dopamine and titania.Lipase was covalently attached on the functionalized titania surface using glutaraldehyde as the cross linking agent.The loading ratio and relative activity of the immobilized lipase were 230 mg/g titania submicrospheres and 65%,respectively.The kinetic parameters including the Michaelis constant (Km) and the maximum reaction rate (Vmax) changed slightly after immobilization.Compared to free lipase,the immobilized lipase showed favorable pH stability,thermostability,recycling stability and storage stability.The immobilized lipase retained 90% activity after incubation at 50 ℃ for 2 h,while the free lipase retained only 60% activity.The immobilized lipase retained more than 80% activity after 8 batches.

  20. Lipoprotein Lipase mRNA expression in different tissues of farm ...

    African Journals Online (AJOL)

    Lipoprotein Lipase mRNA expression in different tissues of farm animals. ... Lipoprotein lipase (LPL) controls triacylglycerol partitioning between adipose tissues and muscles, so it is important enzyme for ... Article Metrics. Metrics Loading .

  1. Hindiii and S447x polymorphisms of lipoprotein lipase gene and ...

    African Journals Online (AJOL)

    Hindiii and S447x polymorphisms of lipoprotein lipase gene and their relationship to coronary artery disease. ... Lipoprotein lipase is a key enzyme in lipoprotein metabolism and its gene is a major candidate gene for coronary ... Article Metrics.

  2. The effect of interaction between Lipoprotein Lipase and ApoVLDL-II ...

    African Journals Online (AJOL)

    The effect of interaction between Lipoprotein Lipase and ApoVLDL-II genes on fat and serum biochemical levels. ... Single nucleotide polymorphism (SNP) in apoVLDL-II and lipoprotein lipase genes was screened by ... Article Metrics.

  3. Biochemical characterization of the surface-associated lipase of Staphylococcus saprophyticus.

    Science.gov (United States)

    Sakinç, Türkân; Kleine, Britta; Gatermann, Sören G

    2007-09-01

    Staphylococcus saprophyticus, an important cause of urinary tract infections, produces a surface-associated lipase, Ssp. In contrast to other lipases, Ssp is a protein that is present in high amounts on the surface of the bacteria and it was shown that it is a true lipase. Characterization of S. saprophyticus lipase (Ssp) showed that it is more similar to Staphylococcus aureus lipase and Staphylococcus epidermidis lipase than to Staphylococcus hyicus lipase and Staphylococcus simulans lipase. Ssp showed an optimum of lipolytic activity at pH 6 and lost its activity at pH>8 or pH<5. The present results show that Ssp activity is dependent on Ca(2+). Consequently, activity increased c. 10-fold in the presence of 2 mM Ca(2+). Optimal activity was reached at 30 degrees C. It was also observed that the enzymatic activity of Ssp depends strongly on the acyl chain length of the substrate molecule.

  4. Bacterial biocatalysts : Molecular Biology, Three-Dimensional Structures, and Biotechnological Applications of Lipases

    NARCIS (Netherlands)

    Jaeger, K-E.; Dijkstra, B.W.; Reetz, M.T.

    1999-01-01

    Bacteria produce and secrete lipases, which can catalyze both the hydrolysis and the synthesis of long-chain acylglycerols. These reactions usually proceed with high regioselectivity and enantioselectivity, and, therefore, lipases have become very important stereoselective biocatalysts used in organ

  5. NEW LIPASE-PRODUCERS MICROORGANISMS FROM PERUVIAN AMAZONIA WHICH HYDROLYZE PALM OIL AND DERIVATIVES

    OpenAIRE

    Roxana Trujillo; Pedro Peláez; Josep-Vicent Sinisterra

    2014-01-01

    Two yeasts: Cryptococcus uchicensis TMY9 and Pichia uchicensis TMY10 and one fungus Verticillium tingalensis TMFMB are described for the first time as lipase producer microorganisms. The strains have been isolated after an ecological screening in a palm oil industry. The yeasts- C. uchicensis and Pichia uchicensis - mainly produce extracellular lipases as active as those produced by traditional lipase producing microorganisms. The extracellular lipases are active in the hydrolysis of crude pa...

  6. Three-Dimensional Structure of Arabidopsis thaliana Lipase Predicted by Homology Modeling Method

    OpenAIRE

    2011-01-01

    Triacylglycerol lipases have been thoroughly characterized in mammals and microorganisms. By contrast, very little is known about plant lipases. In this investigation, a homology model of Arabidopsis thaliana lipase (NP_179126) was constructed using a human gastric lipase (PDB ID: 1HLG), as a template for model building. This model was then assessed for stereochemical quality and side chain environment. Natural substrates: tributyrin, trioctanoin and triolen were docked into the model to inve...

  7. Anaerobic biodegradability of dairy wastewater pretreated with porcine pancreas lipase

    Directory of Open Access Journals (Sweden)

    Adriano Aguiar Mendes

    2010-12-01

    Full Text Available Lipids-rich wastewater was partial hydrolyzed with porcine pancreas lipase and the efficiency of the enzymatic pretreatment was verified by the comparative biodegradability tests (crude and treated wastewater. Alternatively, simultaneous run was carried out in which hydrolysis and digestion was performed in the same reactor. Wastewater from dairy industries and low cost lipase preparation at two concentrations (0.05 and 0.5% w.v-1 were used. All the samples pretreated with enzyme showed a positive effect on organic matter removal (Chemical Oxygen Demand-COD and formation of methane. The best results were obtained when hydrolysis and biodegradation were performed simultaneously, attaining high COD and color removal independent of the lipase concentration. The enzymatic treatment considerably improved the anaerobic operational conditions and the effluent quality (lower content of suspended solids and less turbidity. Thus, the use of enzymes such as lipase seemed to be a very promising alternative for treating the wastewaters having high fat and grease contents, such as those from the dairy industry.O presente trabalho teve como objetivo o pré-tratamento de efluente da indústria de laticínios na hidrólise de lipídeos, empregando lipase de fonte de células animais de baixo custo disponível comercialmente (pâncreas de porco na formação de gás metano por biodegradabilidade anaeróbia empregando diferentes concentrações de lipase (0,05 e 0,5 % w.v-1. A utilização de lipase no pré-tratamento do efluente acelerou a hidrólise de lipídeos e, conseqüentemente, auxiliou o tratamento biológico resultando na redução da matéria orgânica em termos de Demanda Química de Oxigênio (DQO, cor e sólidos em suspensão como lipídeos. Os melhores resultados em termos de remoção de DQO e cor foram obtidos quando a hidrólise e biodigestão foram realizadas simultaneamente, independente da concentração de lipase empregada. Estes resultados

  8. Estolides synthesis catalyzed by immobilized lipases.

    Science.gov (United States)

    Aguieiras, Erika C G; Veloso, Cláudia O; Bevilaqua, Juliana V; Rosas, Danielle O; da Silva, Mônica A P; Langone, Marta A P

    2011-01-01

    Estolides are vegetable-oil-based lubricants obtained from oleic acid or any source of hydroxy fatty acids. In this work, the estolides synthesis from oleic acid and methyl ricinoleate (biodiesel from castor oil), using immobilized commercial lipases (Novozym 435, Lipozyme RM-IM, and Lipozyme TL-IM) in a solvent-free medium was investigated. Acid value was used to monitor the reaction progress by determining the consumption of acid present in the medium. Novozym 435 showed the best performance. Water removal improved the conversion. Novozym 435 was more active at atmospheric pressure. Novozym 435 was reused four times with conversion reaching 15% after the fourth reaction at 80°C. Estolides produced under the reaction conditions used in this work presented good properties, such as, low temperature properties as pour point (-24°C), viscosity (23.9 cSt at 40°C and 5.2 cSt at 100°C), and viscosity index (153).

  9. Estolides Synthesis Catalyzed by Immobilized Lipases

    Directory of Open Access Journals (Sweden)

    Erika C. G. Aguieiras

    2011-01-01

    Full Text Available Estolides are vegetable-oil-based lubricants obtained from oleic acid or any source of hydroxy fatty acids. In this work, the estolides synthesis from oleic acid and methyl ricinoleate (biodiesel from castor oil, using immobilized commercial lipases (Novozym 435, Lipozyme RM-IM, and Lipozyme TL-IM in a solvent-free medium was investigated. Acid value was used to monitor the reaction progress by determining the consumption of acid present in the medium. Novozym 435 showed the best performance. Water removal improved the conversion. Novozym 435 was more active at atmospheric pressure. Novozym 435 was reused four times with conversion reaching 15% after the fourth reaction at 80°C. Estolides produced under the reaction conditions used in this work presented good properties, such as, low temperature properties as pour point (−24°C, viscosity (23.9 cSt at 40°C and 5.2 cSt at 100°C, and viscosity index (153.

  10. Application of lipase technology for transesterification of fatty acid ester

    Directory of Open Access Journals (Sweden)

    JOKO SULISTYO

    2005-07-01

    Full Text Available We have reported the potency of microbial extracellular enzyme for synthesis of fatty acid ester. Further investigation was aimed to study capacity of the enzyme on bioprocess of crude palm oil by transesterification of saturated fatty acid to fatty acid ester. We have studied some lipases from culture filtrate of Candida rugosa FM-9301, Bacillus subtilis FM-9101 and Pseudomonas aerogenes FM-9201, which were preincubated in a medium containing olive oil as inducers, using a shaker under conditions that allowed for lipase production at pH 4.5-6.5 and room temperature for 5 days. Those strains shown different activities during the hydrolysis of substrates, which resulted in decreasing or increasing free fatty acids those, were liberated from media containing crude palm oil and organic solvents. The optimal transesterification condition was at temperature of 45-50C and at pH 4.5 for C. rugosa and pH 6.0 to 7.0 for P. aerogenes and B. subtilis. Under the enzyme concentration of 50% (v/v, the transesterification was rapidly occurred, while at the concentration of 20% (v/v the enzymatically biosynthesis required longer incubation period. The substrates incubated with C. rugosa lipase exhibited higher linoleic and linolenic acid (7.16 and 2.15%, respectively, than that of B. subtilis lipase (4.85% and 1.43%, respectively, while P. aerogenes lipase (3.73% and 1.11%, respectively.

  11. Genome shuffling enhances lipase production of thermophilic Geobacillus sp.

    Science.gov (United States)

    Chalopagorn, Pornchanok; Charoenpanich, Jittima; Choowongkomon, Kiattawee

    2014-10-01

    Thermostable lipases are potential enzymes for biocatalytic application. In this study, the lipase production of Geobacillus sp. CF03 (WT) was improved by genome shuffling. After two rounds of genome shuffling, one fusant strain (FB1) achieved increase lipase activity from the populations generated by ultraviolet irradiation and ethyl methylsulfonate (EMS) mutagenesis. The growth rate and lipase production of FB1 increased highest by 150 and 238 %, respectively, in comparison to the wild type. The fusant enzyme had a significant change in substrate specificity but still prefers the long-chain length substrates. It had an optimum activity at 60 °C, pH at 7.0-8.0, with p-nitrophenyl palmitate (C16) as a substrate and retained about 50 % of their activity after 15 min at 70 °C, pH 8.0. Furthermore, the fusant lipase showed the preference of sesame oil, waste palm oil, and canola oil. Therefore, the genome shuffling strategy has been successful to strain improvement and selecting strain with multiple desirable characteristics.

  12. Lipase-catalyzed polyester synthesis--a green polymer chemistry.

    Science.gov (United States)

    Kobayashi, Shiro

    2010-01-01

    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 chemoenzymatic 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'.

  13. Exploring the conformational states and rearrangements of Yarrowia lipolytica Lipase.

    Science.gov (United States)

    Bordes, Florence; Barbe, Sophie; Escalier, Pierre; Mourey, Lionel; André, Isabelle; Marty, Alain; Tranier, Samuel

    2010-10-06

    We report the 1.7 Å resolution crystal structure of the Lip2 lipase from Yarrowia lipolytica in its closed conformation. The Lip2 structure is highly homologous to known structures of the fungal lipase family (Thermomyces lanuginosa, Rhizopus niveus, and Rhizomucor miehei lipases). However, it also presents some unique features that are described and discussed here in detail. Structural differences, in particular in the conformation adopted by the so-called lid subdomain, suggest that the opening mechanism of Lip2 may differ from that of other fungal lipases. Because the catalytic activity of lipases is strongly dependent on structural rearrangement of this mobile subdomain, we focused on elucidating the molecular mechanism of lid motion. Using the x-ray structure of Lip2, we carried out extensive molecular-dynamics simulations in explicit solvent environments (water and water/octane interface) to characterize the major structural rearrangements that the lid undergoes under the influence of solvent or upon substrate binding. Overall, our results suggest a two-step opening mechanism that gives rise first to a semi-open conformation upon adsorption of the protein at the water/organic solvent interface, followed by a further opening of the lid upon substrate binding.

  14. Identification and characterization of a lipase gene from Antrodia cinnamomea.

    Science.gov (United States)

    Chu, Fang-Hua; Wang, Sheng-Yang; Lee, Li-Chiun; Shaw, Jei-Fu

    2008-12-01

    A partial (634 bp) cDNA clone, AF1229, obtained from expressed sequence tags (ESTs) of solid-cultured basidiomes of Antrodia cinnamomea is homologous to the lipase gene in Rhizomucor miehei. 5'-rapid amplification of cDNA ends (RACE) and 3'-RACE amplification showed that the full-length lipase gene, Ac-LIP, has a 912bp open reading frame (ORF), a 183bp 5' non-coding region, and a 144bp 3' non-coding region. Ac-LIP contains the lipase consensus sequence, VTVVGHSLGA, and encodes a 303-amino acid polypeptide that appears to be an extracellular protein with a calculated molecular mass of 31.8 kDa. RT-PCR analysis suggested that Ac-LIP was strongly expressed during the basidiomatal formation stage of A. cinnamomea. When over-expressed in Escherichia coli, Ac-LIP yielded a protein that was capable of performing hydrolysis of trilinolein by gas chromatography/mass spectrometry (GC/MS) analysis. A. cinnamomea lipase represents the first enzyme of the lipase family from a basidiomycetous fungus, which has been characterized at the molecular level.

  15. Lipase-catalyzed polyester synthesis – A green polymer chemistry

    Science.gov (United States)

    Kobayashi, Shiro

    2010-01-01

    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’. PMID:20431260

  16. Effect of fermentation conditions on lipase production by Candida utilis

    Directory of Open Access Journals (Sweden)

    SANJA Z. GRBAVCIC

    2007-08-01

    Full Text Available A wild yeast strain isolated from spoiled soybean oil and identified as Candida utilis initially presented rather low lipase activity (approximately 4 IU dm-3 in submerged culture in a universal yeast medium containing 2 % malt extract. Stu­dies were undertaken to improve the lipase production. The best yields of lipase were obtained with a medium supplemented with caprylic and oleic acids as indu­cers, but higher concentrations of the former (> 0.5 % had a negative effect on the lipase production and cell growth. The type of nitrogen source seemed also to be very important. The highest lipolytic activity of 284 IU dm-3 was achieved after 5 days of fermentation in a medium containing oleic acid and hydrolyzed casein as carbon and nitrogen sources, respectively, and supplemented with Tween 80®. It was shown that optimization of the fermentation conditions can lead to a significant improvement in the lipase production (more than 70-fold higher compared to the initial value obtained in the non-optimized medium.

  17. Postheparin plasma lipases and carnitine in infants during parenteral nutrition.

    Science.gov (United States)

    Rovamo, L

    1985-03-01

    Lipoprotein lipase is the rate-limiting factor for hydrolyzing triglycerides to glycerol and fatty acids. Carnitine is a cofactor in the transport of long-chain fatty acids through the mitochondrial membrane for oxidation. To assess these determinants of fat utilization during total parenteral nutrition, lipoprotein and hepatic lipase activities and carnitine concentrations of nine newborn infants, operated on because of gastrointestinal anomalies during the first day of life, were measured with specific methods. Total parenteral nutrition was built up in 3 days whereafter the infants received 3 g/kg of fat at a constant rate of infusion for 24 h/day. Lipoprotein lipase activity of post-heparin plasma increased from 14 to 35 mumol free fatty acids/ml/h during parenteral nutrition whereas hepatic lipase activity remained unchanged at 40 mumol free fatty acids/ml/h. Serum free carnitine and acylcarnitine levels decreased significantly during parenteral nutrition; urinary excretion of carnitine decreased also. In addition, serum cholesterol and phospholipids increased markedly during parenteral nutrition whereas serum triglycerides, free fatty acids, and blood beta-hydroxybutyrate remained unchanged. Serum apolipoprotein A-I concentrations were unaltered, apolipoprotein A-II underwent a transient increase, and apolipoprotein B increased monotonically during parenteral nutrition. The results suggest that under the present circumstances neither lipoprotein lipase activity nor carnitine resources are rate-limiting for the utilization of fat in newborn infants during total parenteral nutrition.

  18. FNDC5 overexpression and irisin ameliorate glucose/lipid metabolic derangements and enhance lipolysis in obesity.

    Science.gov (United States)

    Xiong, Xiao-Qing; Chen, Dan; Sun, Hai-Jian; Ding, Lei; Wang, Jue-Jin; Chen, Qi; Li, Yue-Hua; Zhou, Ye-Bo; Han, Ying; Zhang, Feng; Gao, Xing-Ya; Kang, Yu-Ming; Zhu, Guo-Qing

    2015-09-01

    Irisin is a cleaved and secreted fragment of fibronectin type III domain containing 5 (FNDC5), and contributes to the beneficial effects of exercise on metabolism. Here we report the therapeutical effects of FNDC5/irisin on metabolic derangements and insulin resistance in obesity, and show the lipolysis effect of irisin and its signal molecular mechanism. In obese mice, lentivirus mediated-FNDC5 overexpression enhanced energy expenditure, lipolysis and insulin sensitivity, and reduced hyperlipidemia, hyperglycemia, hyperinsulinism, blood pressure and norepinephrine levels; it increased hormone-sensitive lipase (HSL) expression and phosphorylation, and reduced perilipin level and adipocyte diameter in adipose tissues. Subcutaneous perfusion of irisin reduced hyperlipidemia and hyperglycemia, and improved insulin resistance. Either FNDC5 overexpression or irisin perfusion only induced a tendency toward a slight decrease in body weight in obese mice. In 3T3-L1 adipocytes, irisin enhanced basal lipolysis rather than isoproterenol-induced lipolysis, which were prevented by inhibition of adenylate cyclase or PKA; irisin increased the HSL and perilipin phosphorylation; it increased PKA activity, and cAMP and HSL mRNA levels, but reduced perilipin expression. These results indicate that FNDC5/irisin ameliorates glucose/lipid metabolic derangements and insulin resistance in obese mice, and enhances lipolysis via cAMP-PKA-HSL/perilipin pathway. FNDC5 or irisin can be taken as an effective therapeutic strategy for metabolic disorders. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. Acetylshikonin from Zicao Prevents Obesity in Rats on a High-Fat Diet by Inhibiting Lipid Accumulation and Inducing Lipolysis.

    Directory of Open Access Journals (Sweden)

    Meiling Su

    Full Text Available Various drugs have been developed to treat obesity, but these have undesirable secondary effects, and an efficient but non-toxic anti-obesity drug from natural sources is desired. This study investigated the anti-obesity effects and mechanisms of action of acetylshikonin (AS-which is used in traditional Chinese medicine-in rats on a high-fat diet (HFD. Rats were fed a normal diet or an HFD; the latter group was received no treatment or were treated with 100, 300, or 900 mg/kg AS extract by intragastric administration for 6 weeks. In addition, 3T3-L1 adipocytes were treated with AS and the effects on adipogenesis and lipolysis were evaluated by western blot analysis of adipogenic transcription factors and lipid-metabolizing enzyme levels and the phosphorylation status of protein kinase (PK A and hormone-sensitive lipase (HSL. AS prevented HFD-induced obesity including reduction in body weight, white adipose tissue content, liver mass, and serum triglyceride and free fatty acid levels in rats. It also suppressed the expression of adipogenic differentiation transcription factors and decreased the expression of the adipocyte-specific proteins HSL and adipose triglyceride lipase (ATGL. Furthermore, AS treatment induced lipolysis, leading to the release of glycerol and increased in PKA and HSL phosphorylation. These findings demonstrate that AS has anti-obesity effects in a rat model and may be a safe treatment for obesity in humans.

  20. Acetylshikonin from Zicao Prevents Obesity in Rats on a High-Fat Diet by Inhibiting Lipid Accumulation and Inducing Lipolysis.

    Science.gov (United States)

    Su, Meiling; Huang, Wendong; Zhu, Banghao

    2016-01-01

    Various drugs have been developed to treat obesity, but these have undesirable secondary effects, and an efficient but non-toxic anti-obesity drug from natural sources is desired. This study investigated the anti-obesity effects and mechanisms of action of acetylshikonin (AS)-which is used in traditional Chinese medicine-in rats on a high-fat diet (HFD). Rats were fed a normal diet or an HFD; the latter group was received no treatment or were treated with 100, 300, or 900 mg/kg AS extract by intragastric administration for 6 weeks. In addition, 3T3-L1 adipocytes were treated with AS and the effects on adipogenesis and lipolysis were evaluated by western blot analysis of adipogenic transcription factors and lipid-metabolizing enzyme levels and the phosphorylation status of protein kinase (PK) A and hormone-sensitive lipase (HSL). AS prevented HFD-induced obesity including reduction in body weight, white adipose tissue content, liver mass, and serum triglyceride and free fatty acid levels in rats. It also suppressed the expression of adipogenic differentiation transcription factors and decreased the expression of the adipocyte-specific proteins HSL and adipose triglyceride lipase (ATGL). Furthermore, AS treatment induced lipolysis, leading to the release of glycerol and increased in PKA and HSL phosphorylation. These findings demonstrate that AS has anti-obesity effects in a rat model and may be a safe treatment for obesity in humans.

  1. High cell density fed-batch fermentations for lipase production: feeding strategies and oxygen transfer.

    Science.gov (United States)

    Salehmin, M N I; Annuar, M S M; Chisti, Y

    2013-11-01

    This review is focused on the production of microbial lipases by high cell density fermentation. Lipases are among the most widely used of the enzyme catalysts. Although lipases are produced by animals and plants, industrial lipases are sourced almost exclusively from microorganisms. Many of the commercial lipases are produced using recombinant species. Microbial lipases are mostly produced by batch and fed-batch fermentation. Lipases are generally secreted by the cell into the extracellular environment. Thus, a crude preparation of lipases can be obtained by removing the microbial cells from the fermentation broth. This crude cell-free broth may be further concentrated and used as is, or lipases may be purified from it to various levels. For many large volume applications, lipases must be produced at extremely low cost. High cell density fermentation is a promising method for low-cost production: it allows a high concentration of the biomass and the enzyme to be attained rapidly and this eases the downstream recovery of the enzyme. High density fermentation enhances enzyme productivity compared with the traditional submerged culture batch fermentation. In production of enzymes, a high cell density is generally achieved through fed-batch operation, not through perfusion culture which is cumbersome. The feeding strategies used in fed-batch fermentations for producing lipases and the implications of these strategies are discussed. Most lipase-producing microbial fermentations require oxygen. Oxygen transfer in such fermentations is discussed.

  2. The structure-function relationship of the lipases from Pseudomonas aeruginosa and Bacillus subtilis

    NARCIS (Netherlands)

    Misset, Onno; Gerritse, Gijs; Jaeger, Karl-Erich; Winkler, Ulrich; Colson, Charles; Schanck, Karin; Lesuisse, Emmanuel; Dartois, Véronique; Blaauw, Mieke; Ransac, Stéphane; Dijkstra, Bauke W.

    1994-01-01

    Within the BRIDGE T-project on lipases we investigate the structure-function relationships of the lipases from Bacillus subtilis and Pseudomonas aeruginosa. Construction of an overproducing Bacillus strain allowed the purification of > 100 mg lipase from 30 I culture supernatant. After testing a lar

  3. Rat liver contains a limited number of binding sites for hepatic lipase

    NARCIS (Netherlands)

    G.C. Schoonderwoerd (Kees); A.J.M. Verhoeven (Adrie); H. Jansen (Hans)

    1994-01-01

    textabstractThe binding of hepatic lipase to rat liver was studied in an ex vivo perfusion model. The livers were perfused with media containing partially purified rat hepatic lipase or bovine milk lipoprotein lipase. The activity of the enzymes was determined in the pe

  4. 21 CFR 184.1420 - Lipase enzyme preparation derived from Rhizopus niveus.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Lipase enzyme preparation derived from Rhizopus... preparation derived from Rhizopus niveus. (a) Lipase enzyme preparation contains lipase enzyme (CAS Reg. No... nonpathogenic and nontoxigenic strain of Rhizopus niveus. The enzyme preparation also contains...

  5. Green tea catechins enhance norepinephrine-induced lipolysis via a protein kinase A-dependent pathway in adipocytes.

    Science.gov (United States)

    Chen, Shu; Osaki, Noriko; Shimotoyodome, Akira

    2015-05-22

    Green tea catechins have been shown to attenuate obesity in animals and humans. The catechins activate adenosine monophosphate-activated protein kinase (AMPK), and thereby increase fatty acid oxidation in liver and skeletal muscles. Green tea catechins have also been shown to reduce body fat in humans. However, the effect of the catechins on lipolysis in adipose tissue has not been fully understood. The aim of this study was to clarify the effect of green tea catechins on lipolysis in adipocytes and to elucidate the underlying mechanism. Differentiated mouse adipocyte cell line (3T3-L1) was stimulated with green tea catechins in the presence or absence of norepinephrine. Glycerol and free fatty acids in the media were measured. Phosphorylation of hormone-sensitive lipase (HSL) was determined by Western blotting, and the mRNA expression levels of HSL, adipose triglyceride lipase (ATGL), and perilipin were determined by quantitative RT-PCR. The cells were treated with inhibitors of protein kinase A (PKA), protein kinase C (PKC), protein kinase G (PKG), or mitogen-activated protein kinase (MAPK) to determine the responsible pathway. Treatment of 3T3-L1 adipocytes with green tea catechins increased the level of glycerol and free fatty acids released into the media in the presence, but not absence, of norepinephrine, and increased the level of phosphorylated HSL in the cells. The catechins also increased mRNA and protein levels of HSL and ATGL. PKA inhibitor (H89) attenuated the catechin-induced increase in glycerol release and HSL phosphorylation. The results demonstrate that green tea catechins enhance lipolysis in the presence of norepinephrine via a PKA-dependent pathway in 3T3-L1 adipocytes, providing a potential mechanism by which green tea catechins could reduce body fat.

  6. Taking control over intracellular fatty acid levels is essential for the analysis of thermogenic function in cultured primary brown and brite/beige adipocytes.

    Science.gov (United States)

    Li, Yongguo; Fromme, Tobias; Schweizer, Sabine; Schöttl, Theresa; Klingenspor, Martin

    2014-10-01

    Thermogenesis in brown adipocytes, conferred by mitochondrial uncoupling protein 1 (UCP1), is receiving great attention because metabolically active brown adipose tissue may protect humans from metabolic diseases. In particular, the thermogenic function of brown-like adipocytes in white adipose tissue, known as brite (or beige) adipocytes, is currently of prime interest. A valid procedure to quantify the specific contribution of UCP1 to thermogenesis is thus of vital importance. Adrenergic stimulation of lipolysis is a common way to activate UCP1. We here report, however, that in this frequently applied setup, taking control over intracellular fatty acid levels is essential for the analysis of thermogenic function in cultured brown and brite adipocytes. By the application of these findings, we demonstrate that UCP1 is functionally thermogenic in intact brite adipocytes and adrenergic UCP1 activation is largely dependent on adipose triglyceride lipase (ATGL) rather than hormone sensitive lipase (HSL).

  7. QSAR study and the hydrolysis activity prediction of three alkaline lipases from different lipase-producing microorganisms.

    Science.gov (United States)

    Wang, Haikuan; Wang, Xiaojie; Li, Xiaolu; Zhang, Yehong; Dai, Yujie; Guo, Changlu; Zheng, Heng

    2012-09-28

    The hydrolysis activities of three alkaline lipases, L-A1, L-A2 and L-A3 secreted by different lipase-producing microorganisms isolated from the Bay of Bohai, P. R. China were characterized with 16 kinds of esters. It was found that all the lipases have the ability to catalyze the hydrolysis of the glycerides, methyl esters, ethyl esters, especially for triglycerides, which shows that they have broad substrate spectra, and this property is very important for them to be used in detergent industry. Three QSAR models were built for L-A1, L-A2 and L-A3 respectively with GFA using Discovery studio 2.1. The models equations 1, 2 and 3 can explain 95.80%, 97.45% and 97.09% of the variances (R(2)(adj)) respectively while they could predict 95.44%, 89.61% and 93.41% of the variances (R(2)(cv)) respectively. With these models the hydrolysis activities of these lipases to mixed esters were predicted and the result showed that the predicted values are in good agreement with the measured values, which indicates that this method can be used as a simple tool to predict the lipase activities for single or mixed esters.

  8. QSAR study and the hydrolysis activity prediction of three alkaline lipases from different lipase-producing microorganisms

    Directory of Open Access Journals (Sweden)

    Wang Haikuan

    2012-09-01

    Full Text Available Abstract The hydrolysis activities of three alkaline lipases, L-A1, L-A2 and L-A3 secreted by different lipase-producing microorganisms isolated from the Bay of Bohai, P. R. China were characterized with 16 kinds of esters. It was found that all the lipases have the ability to catalyze the hydrolysis of the glycerides, methyl esters, ethyl esters, especially for triglycerides, which shows that they have broad substrate spectra, and this property is very important for them to be used in detergent industry. Three QSAR models were built for L-A1, L-A2 and L-A3 respectively with GFA using Discovery studio 2.1. The models equations 1, 2 and 3 can explain 95.80%, 97.45% and 97.09% of the variances (R2adj respectively while they could predict 95.44%, 89.61% and 93.41% of the variances (R2cv respectively. With these models the hydrolysis activities of these lipases to mixed esters were predicted and the result showed that the predicted values are in good agreement with the measured values, which indicates that this method can be used as a simple tool to predict the lipase activities for single or mixed esters.

  9. Investigation of the Reuse of Immobilized Lipases in Biodiesel Synthesis: Influence of Different Solvents in Lipase Activity.

    Science.gov (United States)

    Aguieiras, Erika C G; Ribeiro, Douglas S; Couteiro, Pedro P; Bastos, Caenam M B; de Queiroz, Danielle S; Parreira, Juliana M; Langone, Marta A P

    2016-06-01

    Biodiesel production catalyzed by immobilized lipases offers the possibility of easy reuse of the catalyst, which is very important to minimize costs and to make this process economically feasible. In this study, the reuse of three commercial immobilized lipases (Novozym 435, Lipozyme RM IM, and Lipozyme TL IM) was investigated in ethanolysis of soybean oil. The effect of the use of solvents (ethanol, butanol, and hexane) to wash the immobilized lipases before the enzyme reuse was evaluated, as well as the lipase reuse without solvent washing. The washing with butanol and ethanol led to the lowest decrease in ester yield after the first batch and allowed the highest glycerol removal (>85 %) from biocatalysts. The biocatalysts were incubated at 50 °C for 2 h in these three solvents. Esterification activities of the enzyme preparations, scanning electron microscopy (SEM) analyses of the beads, and protein content in organic phase were evaluated before and after incubation in the solvent. SEM analysis showed a significant change in beads morphology of Novozym 435 after contact with hexane. For Lipozyme TL IM lipase, this effect was visualized with ethanol.

  10. Synthesis of Wax Esters by Lipase-catalyzed Esterification with Immobilized Lipase from Candida sp. 99-125

    Institute of Scientific and Technical Information of China (English)

    邓利; 王晓静; 聂开立; 王芳; 刘军峰; 王璞; 谭天伟

    2011-01-01

    Wax esters were synthesized in a solvent free system catalyzed by immobilized lipase from Candida sp. 99-125, with oleic acid and cetyl alcohol. The effects of substrate molar ratio, lipase dosage and water removal were investigated in a 50 ml flask incubated in a thermostatic cultivation cabinet. The optimized conditions were: temperature 40 ℃, shaking at 170 r·min-1, acid/alcohol molar ratio 1:0.9, lipase dosage in 10% (by mass) of oleic acid, and open reaction for water removal. As a result, the conversion rate reached 98% for reaction of 8 h. The volume of reactor was scaled up to 1 L three-neck flask. The optimized parameters were: 200 r·min-1 agitation speed, 2.5% (by mass) lipase dosage, others were the same as the parameters described above. The conversion rate reached 95% for reaction of 24 h. The lipase retained 46% conversion rate after reuse for 6, 7 batches. The products were purified by removing remained cetyl alcohol and fatty acids with ethanol and saturated sodium carbonate so-lution, respectively. The purity of the wax ester, cetyl oleate, was 96%. The physical and chemical properties of cetyl oleate were tested and compared with those of jojoba oil. The results show that the product cetyl oleate has great potential to use as the substitute of natural jojoba oil.

  11. Lipase-catalyzed biodiesel synthesis with different acyl acceptors

    Directory of Open Access Journals (Sweden)

    Ognjanović Nevena D.

    2008-01-01

    Full Text Available Biodiesel is an alternative fuel for diesel engine that is environmentally acceptable. Conventionally, biodiesel is produced by transesterification of triglycerides and short alcohols in the presence of an acid or an alkaline catalyst. There are several problems associated with this kind of production that can be resolved by using lipase as the biocatalyst. The aim of the present work was to investigate novel acyl acceptors for biodiesel production. 2-Propanol and n-butanol have a less negative effect on lipase stability, and they also improve low temperature properties of the fuel. However, excess alcohol leads to inactivation of the enzyme, and glycerol, a major byproduct, can block the immobilized enzyme, resulting in low enzymatic activity. This problem was solved by using methyl acetate as acyl acceptor. Triacetylglycerol is produced instead of glycerol, and it has no negative effect on the activity of the lipase.

  12. Lactobacillus sps. lipase mediated poly (ε-caprolactone) degradation.

    Science.gov (United States)

    Khan, Imran; Ray Dutta, Jayati; Ganesan, Ramakrishnan

    2017-02-01

    Polymer degradation through lipase appears to be an enthralling alternative to bulk chemical routes. Poly (ε-caprolactone) (PCL) is an artificial polyester that can be degraded by microbes and enzymes like lipases and esterases. The environmental degradation of PCL is dependent on the activity of bacteria that characterization techniques such as thermogravimetric analysis, differential thermal are widely present in the ecosystem. In this study, three different lipases derived from Lactobacillus brevis, Lactobacillus plantarum and their co-culture have been utilized to explore their efficiency towards PCL enzymatic degradation. The effect of parameters such as enzyme loading and degradation time has been explored to understand the efficiency of the enzymes used in this study. Various analysis, scanning electron microscopy and Fourier transform infrared spectroscopy have been employed to study the enzymatic degradation and its possible mechanistic insight.

  13. Lipase-immobilized biocatalytic membranes for biodiesel production.

    Science.gov (United States)

    Kuo, Chia-Hung; Peng, Li-Ting; Kan, Shu-Chen; Liu, Yung-Chuan; Shieh, Chwen-Jen

    2013-10-01

    Microbial lipase from Candida rugosa (Amano AY-30) has good transesterification activity and can be used for biodiesel production. In this study, polyvinylidene fluoride (PVDF) membrane was grafted with 1,4-diaminobutane and activated by glutaraldehyde for C. rugosa lipase immobilization. After immobilization, the biocatalytic membrane was used for producing biodiesel from soybean oil and methanol via transesterification. Response Surface Methodology (RSM) in combination with a 5-level-5-factor central composite rotatable design (CCRD) was employed to evaluate the effects of reaction time, reaction temperature, enzyme amount, substrate molar ratio and water content on the yield of soybean oil methyl ester. By ridge max analysis, the predicted and experimental yields under the optimum synthesis conditions were 97% and 95%, respectively. The lipase-immobilized PVDF membrane showed good reuse ability for biodiesel production, enabling operation for at least 165 h during five reuses of the batch, without significant loss of activity.

  14. The immobilization of lipase on PVDF-co-HFP membrane

    Science.gov (United States)

    Kayhan, Naciye; Eyüpoǧlu, Volkan; Adem, Şevki

    2016-04-01

    Lipase is an enzyme having a lot of different industrial applications such as biodiesel production, biopolymer synthesis, enantiopure pharmaceutical productions, agrochemicals, etc. Its immobilized form on different substances is more conventional and useful than its free form. Supporting material was prepared using PVDF-co-HFP in laboratory conditions and attached 1,4-diaminobutane (DA) and epichlorohydrin (EPI) ligands to the membrane to immobilize lipase enzyme. The immobilization conditions such as enzyme amount, pH, the concentration of salt, thermal stability and activity were stabilized for our experimental setup. Then, biochemical characterizations were performed on immobilized lipase PVDF-co-HFP regarding optimal pH activity, temperature and thermal stability. Also, the desorption ratios of immobilized enzyme in two different pathway were investigated to confirm immobilization stability for 24 hours.

  15. Biodiesel production from microalgae oil catalyzed by a recombinant lipase.

    Science.gov (United States)

    Huang, Jinjin; Xia, Ji; Jiang, Wei; Li, Ying; Li, Jilun

    2015-03-01

    A recombinant Rhizomucor miehei lipase was constructed and expressed in Pichia pastoris. The target enzyme was termed Lipase GH2 and it can be used as a free enzyme for catalytic conversion of microalgae oil mixed with methanol or ethanol for biodiesel production in an n-hexane solvent system. Conversion rates of two major types of biodiesel, fatty acid methyl ester (FAME) and fatty acid ethyl ester (FAEE), reached maximal values (>90%) after 24h. The process of FAME production is generally more simple and economical than that of FAEE production, even though the two processes show similar conversion rates. In spite of the damaging effect of ethanol on enzyme activity, we successfully obtained ethyl ester by the enzymatic method. Our 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.

  16. Effects of temperature and pressure on Rhizomucor miehei lipase stability.

    Science.gov (United States)

    Noel, Marilyne; Combes, Didier

    2003-04-10

    Both high temperature and high hydrostatic pressure induce irreversible deactivation of enzymes. They enable the enzyme's thermodynamic parameters to be determined and are used to study the mechanisms involved in biochemical systems. The effect of these two factors on the stability of Rhizomucor miehei lipase have been investigated. The stability criterion used was residual hydrolytic activity of the lipase. Experimental and theoretical parameters, obtained by linear regression analysis, were compared with theoretical kinetics in order to validate the series-type inactivation model. The lipase of R. miehei was deactivated by either thermal or pressure treatment. Moreover conformational studies made by fluorescence spectroscopy suggest that the conformational changes induced by pressure were different from those induced by temperature. In addition they show that after thermal deactivation there were less intermolecular hydrogen bonded structures formed than was the case for deactivation by high pressure.

  17. [Structure and Activity of Fungal Lipases in Bile Salt Solutions].

    Science.gov (United States)

    Bogdanova, L R; Bakirova, D R; Valiullina, Yu A; Idiyatullin, B Z; Faizullin, D A; Zueva, O S; Zuev, Yu F

    2016-01-01

    The changes in structure and catalytic properties of fungal lipases (Candida rugosa, Rhizomucor miehei, Mucor javanicus) were investigated in micellar solutions of bile salts that differ in hydrophilic-lypophilic balance and reaction medium properties. The methods of circular dichroism and tryptophan fluorescence were applied to estimate the changes in peptide structure within complexes with bile salt micelles. Bile salts do not exert a significant influence on the structure of the enzymes under study: in Rh. miehei and M. javanicus lipases the alpha helix content slightly decreased, the influence of bile salts on the C. rugosa structure was not revealed. Despite negligible structural modifications in the enzymes, in bile salt solutions a considerable change in their catalytic properties was observed: an abrupt decrease in catalytic effectiveness. Substrate-bile salts micelles complex formation was demonstrated by the NMR self-diffusion method. The model of a regulation of fungal lipase activity was proposed.

  18. Alginate-chaperoned facile refolding of Chromobacterium viscosum lipase.

    Science.gov (United States)

    Mondal, Kalyani; Bohidar, Himadri B; Roy, Rajendra P; Gupta, Munishwar N

    2006-05-01

    Urea denatured lipase from Chromobacterium viscosum lipase could be refolded by addition of alginate with high guluronic acid content. The refolded molecule could be recovered by affinity precipitation. This approach resulted in recovery of 80% (of original activity) as compared to classical dilution method which gave only 21% activity recovery. Dynamic light scattering showed that binding required about 45 min and activity data obtained from affinity precipitation experiments indicated that refolding was almost instantaneous after binding. Circular dichroism (CD) and fluorescence data showed that refolded molecule was identical to the native molecule. It also showed that refolding takes place at the binding stage and not at the precipitation stage. Preliminary studies showed that the refolding strategy worked equally well with lipases from wheat germ and porcine pancreas.

  19. Screening of supports for immobilization of commercial porcine pancreatic lipase

    Directory of Open Access Journals (Sweden)

    Robison Scherer

    2011-12-01

    Full Text Available The aim of this work is to report the performance of different supports for the immobilization of commercial porcine pancreatic lipase. The immobilization tests were carried out in several types of Accurel, activated alumina, kaolin, montmorillonite, ion exchange resins and zeolites. The characterization of the supports showed differences in terms of specific area and morphology. The characteristics of the supports influenced the amount of enzyme adsorbed, yield of immobilization and esterification activity of the resulting immobilized catalyst. The clays KSF and natural and pillared montmorillonites presented potential for use as support for lipase immobilization in terms of yield and esterification activity. Yields of immobilization of 76.32 and 52.01% were achieved for clays KSF and natural montmorillonite, respectively. Esterification activities of 754.03, 595.51, 591.88 and 515.71 U.g-1 were obtained for lipases immobilized in Accurel MP-100, Amberlite XAD-2, mordenite and pillared montmorillonite, respectively.

  20. Lipase assay in duodenal juice using a conductimetric method.

    Science.gov (United States)

    Ballot, C; Favre-Bonvin, G; Wallach, J M

    1984-11-15

    Lipase activity in duodenal juice is known to undergo important variations in pathologic states, especially in cases of chronic pancreatitis. Almost all of the current assay methods are based on the measurement of hydrolysis of olive oil or triolein, mainly by potentiometry. As we have developed a conductimetric method for enzyme activity measurements, we have applied it to lipase assay. A higher experimental conductimetric sensitivity is obtained when liberated acids have a short chain (higher limiting equivalent conductivity). We have therefore used triacetin as a substrate and compared out method with potentiometry (pH-stat) and spectrophotometry. The correlation coefficients of both methods with conductimetry were 0.94 and 0.97, respectively, indicating that the conductimetric method may be used for lipase assay in duodenal juice, using triacetin as a substrate.

  1. [Overexpression of Penicillium expansum lipase gene in Pichia pastoris].

    Science.gov (United States)

    Yuan, Cai; Lin, Lin; Shi, Qiao-Qin; Wu, Song-Gang

    2003-03-01

    The alkaline lipase gene of Penicillium expansum (PEL) was coloned into the yeast integrative plasmid pPIC3.5K, which was then transformed into His4 mutant yeast GS115. Recombinant Pichia strains were obtained by minimal olive oil-methanol plates screening and confirmed by PCR. The expression producus of PEL gene was analysis by SDS-PAGE and olive oil plate, the result indicated that PEL gene was functionally overexpressed in Pichia pastoris and up to 95% of the secreted protein. Recombinant lipase had a molecular mass of 28kD, showing a range similar to that of PEL, could hydrolyze olive oil and formed clear halos in the olive oil plates. Four different strategies (different media, pH, glycerol and methanol concentration) were applied to optimize the cultivation conditions, the activity of lipase was up to 260 u/mL under the optimal cultivation conditions. It is pointed out that the absence of the expensive biotin and yeast nitrogen base in the medium increased the lipase production. The possible reason of this result is absence of yeast nitrogen base increased the medium pH during cultivation, and PEL shows a higher stability at this condition. The lipase activity of the supernatant from the culture grown at pH 7 was higher than the one from the culture in the same medium at pH 6.0 is due to the pH stability of PEL too. The results also showed that the methanol and glycerol concentration had a marked effect on the production of lipase.

  2. The specificity of Several Kinds Lipases on n-3 Polyunsaturated Fatty Acids

    Directory of Open Access Journals (Sweden)

    Jenny Elisabeth, T Yuliani, P M Tambunan, J M Purba

    2001-04-01

    Full Text Available Several lipases from microbial and plant, i.e Rhizomucor miehei, Pseudomonas sp., Candida antartica, rice bran, and Carica papaya latex (CPL were examined for synthesis of omega-3 (n-3 PUFA-rich glyceride by hydrolysis and acidolysis reaction. Tuna oil was used in hydrolysis reaction, whereas tuna and palm oils were used as source of triglyceride (TAG molecules and n-3 PUFA concentrate from tuna oil as source of EPA and DHA in acidolysis reaction.For hydrolysis reaction, the rice bran and CPL lipases showed the lowest hydrolytic activity of the tuna oil, whereas the R. miehei lipase showed the highest hydrolytic activity but was unable to hydrolyze EPA and DHA. On the contrary, the C. antartica and Pseudomonas sp. lipases acted stronger on hydrolysis of DHA ester bond than EPA.For acidolysis reaction, all the lipases showed ability to incorporate n-3 PUFA into tuna and palm oils. C. antartica lipase had the maximum DHA incorporation into tuna and palm oils, rice bran lipase had relatively similar ability with R. miehei lipase, and the CPL lipase had the lowest ability. This study proved that rice bran and CPL lipases also had transesterification activity and showed the feasibility of the rice bran lipase to be a biocatalyst for n-3 PUFA-rich glyceride production. Increasing the substrate ratio, of n-3 PUFA concentrate and tuna or palm oil, could increase the EPA and DHA incorporation. The R. miehei, rice bran, and CPL lipases unabled to incorporate DHA into DHA-containing glyceride molecule, whereas C. antartica lipase had the capability in high ratio of n-3 PUFA concentrate to oil. Therefore, the lipases were easier to incorporate n-3 PUFA into palm oil than tuna oil, since the TAG molecules of palm oil was not as complex as tuna oil. It could be suggested that the lipases did not only have acyl chain and positional specificity, but also the whole glyceride structure specificity.

  3. Diagnostic value of post-heparin lipase testing in detecting common genetic variants in the LPL and LIPC genes

    NARCIS (Netherlands)

    M. van Hoek (Mandy); G.M. Dallinga-Thie (Geesje); E.W. Steyerberg (Ewout); E.J.G. Sijbrands (Eric)

    2009-01-01

    textabstractPost-heparin lipoprotein lipase and hepatic lipase activities are used to identify primary disorders of triglyceride and HDL-cholesterol metabolism. Their ability to identify common variants in the lipoprotein lipase (LPL) and hepatic lipase (LIPC) genes is unclear. To investigate the ab

  4. Hyperactivation of Rhizomucor miehei lipase by hydrophobic xerogels.

    Science.gov (United States)

    Aucoin, Marc G; Erhardt, Frank A; Legge, Raymond L

    2004-03-20

    Although a variety of approaches exist for the immobilization of enzymes, the "science" of enzyme immobilization is still in its infancy. In recent years, considerable interest has developed regarding the use of xerogels for enzyme immobilization. There are several advantages to xerogels for enzyme immobilization, including the opportunity to produce them in defined shapes or thin films and the ability to manipulate their physical characteristics (e.g., porosity, hydrophobicity, and optical properties). In this study we examined the effect of xerogel hydrophobicity on the activity of lipase (EC 3.2.2.3) from Rhizomucor miehei. The hydrophobicity of the xerogels was manipulated by generating xerogels with various molar ratios of propyltrimethoxysilane (PTMS) to tetramethoxysilane (TMOS), from 1:1 to 10:1. The belief was that, by increasing the proportion of propyl groups, the hydrophobicity of the resulting xerogel would be increased. Differences in the hydrophobicity of the resulting xerogels were confirmed using water-affinity studies. Two approaches were taken for water-affinity determinations by examining the ability of the xerogels to remove water from air (controlled humidity) and from water-saturated isopropyl ether. Xerogels with higher propyl content showed a reduced affinity for water. A crude lipase preparation from Rhizomucor miehei was then contacted with sized xerogel particulates and the effect of the xerogel on lipase activity was determined. The presence of the xerogel resulted in hyperactivation of the lipase. Analysis of the protein adsorption revealed changes in the profile of proteins adsorbed to the xerogel based on the hydrophobicity of the xerogel. Based on estimations of the specific activity of the hyperactivated lipase, a minimum hyperactivation of 207% was observed. Part of the hyperactivation may be attributable to xerogel-lipase interactions, but also to the adsorption of a component from the crude lipase preparation that may complex

  5. Lipases: particularly effective biocatalysts for cosmetic active ingredients

    Directory of Open Access Journals (Sweden)

    Yvergnaux Florent

    2017-07-01

    Full Text Available Enzymes are the tools of choice in the on-going quest for non-pollutant processes to discover molecules for use in skin products. Amongst these biocatalysts, lipases offer considerable potential in terms of ingredient development and are of interest in skin dermocosmetic formulations possessing sensory or biological activities. Lipases have been studied for around thirty years and, in most cases, these enzymes function under what are deemed to be mild conditions, displaying remarkable efficacy particularly in terms of selectivity. This particularly effective strategy will be illustrated through typical synthesis, demonstrating how ester or amide active ingredients are obtained.

  6. Production, purification and characterization of alkaline lipase from Fusarium oxysporum.

    OpenAIRE

    2006-01-01

    Resumo: Recentemente, a aplicação industrial de lipases microbianas tem sido estendida a muitas áreas, como por exemplo, na modificação de triglicerídeos, síntese de vários compostos de ésteres e detergentes. As lipases podem ser aplicadas na limpeza de maquinários industriais ou em detergentes como sabões em pó na remoção de manchas de lipídeos em tecidos. A linhagem de fungo Fusarium oxysporum 152B foi selecionada entre 216 linhagens de microrganismos isolados de amostras de frutas e solo d...

  7. 基于HSL空间彩色图像多结构元形态边缘检测%Edge detection of color image based on multi-structural elements morphology of HSL space

    Institute of Scientific and Technical Information of China (English)

    王金凤; 焦斌亮

    2011-01-01

    构建了一类在HSL颜色空间基于多结构元彩色形态边缘梯度检测算法实现彩色图像边缘检测新算法,多结构元形态边缘检测有着比单一结构元素形态边缘检测更优越的性能.该方法是把RGB空间的彩色图像转换到HSL空间,并且定义了在HSL空间的彩色形态学基本算子,提出了改进的多结构元彩色形态边缘检测算法.经过大量实验证明,该算法在有噪声的干扰下,比传统的方法能够更好地抑制噪声并提取有用的图像边缘信息,能满足不同的应用需要.%A new method of realizing edge detection for color image is constructed based on the gradient algorithm of multi-structuring elements morphology in HSL space.The ability of edge detection based on multi-structuring elements morphology is superior to single-structuring element.In this method color image of RGB space is converted into image of HSL space, the basic color morphological operators in HSL space are defined and improved multi-structuring elements color morphological operators are proposed.The experiments show that this algorithm can more effectively suppress noise and better extract useful image edge information than traditional methods,and it can meet various requirements in application.

  8. In vitro prevention of chick pancreatic lipase activity by Abroma augusta extract

    Institute of Scientific and Technical Information of China (English)

    Nidhi Gupta; Aditya Ganeshpurkar; Nishikant Jatav; Divya Bansal; Nazneen Dubey

    2012-01-01

    Objective: To investigate chick pancreatic lipase inhibitory activities of the Abroma augusta (A. augusta). Methods: A. augusta was first extracted with methanol and subjected to phytochemical screenings. Quantitative estimation of flavonoids, phenolics and alkaloids was done. Pancreatic lipase from chick pancreas was isolated and used as substrate for anti-lipase studies. Results:A. augusta extract effectively inhibited concentration dependent lipase activity, whereby extract at concentration 100 μg/mL inhibited 88.6% enzyme activity. Conclusions: From these results, it could be concluded that A. augusta can be used as a potential source anti-lipase agents.

  9. Pancreas-specific lipase concentrations and amylase and lipase activities in the peritoneal fluid of dogs with suspected pancreatitis.

    Science.gov (United States)

    Chartier, Marie A; Hill, Steve L; Sunico, Sarena; Suchodolski, Jan S; Robertson, Jane E; Steiner, Joerg M

    2014-09-01

    Diagnosing acute pancreatitis in the dog can be challenging. The aim of this study was to determine the concentrations of pancreas-specific lipase immunoreactivity (cPLI), and the activities of amylase and lipase, in the peritoneal fluid from a population of dogs diagnosed with acute pancreatitis based on clinical signs, ultrasonographic findings and serum cPLI concentrations. In a prospective study, cPLI concentrations, and amylase and lipase activities, were measured in the peritoneal fluid of 14 dogs with pancreatitis and 19 dogs with non-pancreatic disease. The sensitivity and specificity of peritoneal fluid cPLI concentration (cut-off value 500 µg/L) were 100.0% (95% confidence interval, CI, 80.7-100.0%) and 94.7% (95% CI 76.7-99.7%), respectively. The sensitivity and specificity of peritoneal fluid amylase (cut-off value 1050 U/L) and lipase activities (cut-off value 500 U/L) were 71.4% (95% CI 44.5-90.2%) and 84.2% (95% CI 62.8-95.8%) for amylase activity, and 92.9% (95% CI 69.5-99.6%) and 94.7% (95% CI 76.7-99.7%) for lipase activity, respectively. In conclusion, peritoneal fluid cPLI concentration was highly sensitive as a complementary diagnostic tool in a group of dogs with suspected acute pancreatitis. Peritoneal fluid lipase activity was not as sensitive as cPLI concentration, but may also support a diagnosis of acute pancreatitis in dogs. Published by Elsevier Ltd.

  10. SECRETION OF LIPASES IN THE DIGESTIVE TRACT OF THE CRICKET Gryllus bimaculatus.

    Science.gov (United States)

    Weidlich, Sandy; Hoffmann, Klaus H; Woodring, Joseph

    2015-12-01

    Little is known concerning the sites and the ratios of the lipase secretions in insects, therefore we undertook an examination of the lipase secretion of fed and unfed adult female Gryllus bimaculatus. The ratio of triacylglyceride lipase, diacylglyceride lipase, and phosphatidylcholine lipase secreted by fed females in the caecum and ventriculus is 1:1.4:0.4. These activities decrease in the caecum by 30-40% in unfed females. The total lipase activity (TLA) in the caecum is about 10 times that in the ventriculus. Minimal lipase secretion occurs before and during the final moult, and remains at this level in unfed crickets, indicating a basal secretion rate. In 2-day-old fed females, about 10% of the TLA in the entire gut is found in the crop, about 70% in the caecum, 20% in the ventriculus, and 3% in the ileum. Lipases in the ventriculus are recycled back to the caecum and little is lost in the feces. Oleic acid stimulated in vitro lipase secretion, but lipids did not. Feeding stimulated lipase secretion, starvation reduced lipase secretion, but this does not prove a direct prandal regulation of secretion, because feeding also induced a size and volume increase of the caecum. © 2015 Wiley Periodicals, Inc.

  11. Structural basis for the cold adaptation of psychrophilic M37 lipase from Photobacterium lipolyticum.

    Science.gov (United States)

    Jung, Suk-Kyeong; Jeong, Dae Gwin; Lee, Mi Sook; Lee, Jung-Kee; Kim, Hyung-Kwoun; Ryu, Seong Eon; Park, Byoung Chul; Kim, Jae Hoon; Kim, Seung Jun

    2008-04-01

    The M37 lipase from Photobacterium lipolyticum shows an extremely low activation energy and strong activity at low temperatures, with optimum activity seen at 298 K and more than 75% of the optimum activity retained down to 278 K. Though the M37 lipase is most closely related to the filamentous fungal lipase, Rhizomucor miehei lipase (RML) at the primary structure level, their activity characteristics are completely different. In an effort to identify structural components of cold adaptation in lipases, we determined the crystal structure of the M37 lipase at 2.2 A resolution and compared it to that of nonadapted RML. Structural analysis revealed that M37 lipase adopted a folding pattern similar to that observed for other lipase structures. However, comparison with RML revealed that the region beneath the lid of the M37 lipase included a significant and unique cavity that would be occupied by a lid helix upon substrate binding. In addition, the oxyanion hole was much wider in M37 lipase than RML. We propose that these distinct structural characteristics of M37 lipase may facilitate the lateral movement of the helical lid and subsequent substrate hydrolysis, which might explain its low activation energy and high activity at low temperatures.

  12. Production of a novel cold-active lipase from Pichia lynferdii Y-7723.

    Science.gov (United States)

    Kim, Hak-Ryul; Kim, In-Hwan; Hou, Ching T; Kwon, Kwang-Il; Shin, Beom-Soo

    2010-01-27

    Lipase (triacylglycerol acylhydrolases, E.C. 3.1.1.3) is one of the most important enzymes applied to a broad range of industrial application fields. Especially, lipases with abnormal functionality such as thermostability and alkaline, acidic, and cold activities gain special attention because of their applicability in the restricted reaction conditions. In this study, 16 yeast strains prescreened for lipase induction were investigated for their actual lipase production, and we found a novel cold-active lipase produced from Pichia lynferdii Y-7723. The activity of lipase Y-7723 was retained by 74 and 70% at 20 and 10 degrees C, respectively, as compared to the maximum value at 35 degrees C. On the basis of an optimization study, the optimal lipase productivity was obtained at 96 h of incubation with 3% oil substrate in a medium composed of sucrose as a carbon source at pH 7.0. Among carbon sources tested, sucrose showed almost twice as high of a lipase production (184%) as the control, while the cell growth was similar (105%). Yeast extract and ammonium salts were effective as individual nitrogen sources for lipase production. This study demonstrated that the cold activity of lipase Y-7723 at 10 degrees C was highest among the cold-active lipases reported so far.

  13. Fermentation Performance and Characterization of Cold-Adapted Lipase Produced with Pseudomonas Lip35

    Institute of Scientific and Technical Information of China (English)

    YU Hong-wei; HAN Jun; LI Ning; QIE Xiao-sha; JIA Ying-min

    2009-01-01

    Strain of Pseudomonas Lip35 producing lipase was isolated in a refrigerator. Lipase production and characterization of this strain were investigated under different conditions. The Pseudomonas was cultivated in shaking flasks in a fermentation medium in various nutritional and physical environments. Lipase production has been influenced by the presence of yeast-extract, soybean powder, NaCl, and Tween-80. Maximum lipase productivity was obtained when the physical environment of the fermentation medium was optimal for 67 h. The production of lipase reached 58.9 U mL-1 The lipase of Pseudomonas Lip35 can be considered to be inducible, but the inducer had little influence on the production of lipase.The lipase was characterized and showed high lipolytic activity from pH 7.5-8.0. The optimum temperature was observed at 20℃ and the thermal inactivation of lipase was obvious at 60℃. The lipase activity was inhibited by K+, stimulated by Ca2+, and thermostability decreased in the presence of Ca2+, therefore the lipase was Ca2+-dependent cold-adapted enzyme.

  14. PPARγ as a sensor of lipase activity and a target for the lipase inhibitor orlistat.

    Science.gov (United States)

    Martin, Harry; McGhie, Tony K; Bentley-Hewitt, Kerry; Christeller, John

    2013-04-08

    A PPARγ fluorescence polarization (FP) assay was used to measure the release of fatty acid products from triglyceride emulsions during digestion with pancreatic and yeast lipases in a real-time, homogenous assay. Using the same FP assay we show the anti-obesity drug Orlistat is a PPARγ ligand with an IC50 of 2.84 ± 0.16 μM. Analytical Mass Spectrometry confirms that Orlistat does not bind covalently to PPARγ. The PPARγ FP assay is shown to be a simple method for measuring real-time lipase activity using a number of triglyceride substrates including olive oil and grape seed oil emulsions. Incubation of Orlistat with the human intestinal epithelial cell line Caco-2, at concentrations of 1 - 100 μM, leads to induction of genes regulated by PPARγ. At 100 μM Orlistat, transcription of β-defensin 1 (hDB1) & Adipose Differentiation Related Protein (ADRP) increase by up to 2.6 fold and 6.8 fold, respectively. Although at 1 μM and 100 μM Orlistat did not significantly increase defensin protein synthesis, at 10 μM Orlistat induced a 1.5 fold increase in hDB1 protein secretion in the human colonic adenocarcinoma cell line HT-29. Thus Orlistat is similar to the anti-diabetic drug Rosiglitazone in its ability to induce defensin gene expression. The antimicrobial peptide β-defensin 1 protects against pathogenic micro-organisms in the gut and PPARγ suppresses inflammatory gene expression. These may be beneficial side effects of Orlistat consumption on gut epithelial cells.

  15. Immobilization of Lipase from Candida Rugosa on Palm-Based Polyurethane Foam as a Support Material

    Directory of Open Access Journals (Sweden)

    Roila Awang

    2007-01-01

    Full Text Available Lipase from Candida rugosa was immobilized onto palm-based polyurethane foam (PU, which the polymer was pre-soaked in co-immobilized agent. The activities of PU-immobilized lipase were tested by the esterification reaction of oleic acid and oleyl alcohol in hexane. The PU-immobilized lipase was then characterized in term of its thermal, operational and storage stability. The optimum temperature for native and PU-immobilized lipase was at 40oC. This shows that the immobilization did not alter the general character of the lipase. The PU-immobilized lipase shows different enzymatic characteristic-incubation time, enzyme concentration, solvent and operational stability compared to Lipozyme IM. The reuse stability of PU-immobilized lipase was at least four cycles. The % conversion above 80% was still achieved for the sample stored at -5oC after 9 days storage.

  16. Immobilization of lipases in PSS/PEO blends and applications in esters synthesis; Imobilizacao de lipases em blendas de PSS/PEO e aplicacoes na sintese de esteres

    Energy Technology Data Exchange (ETDEWEB)

    Vecchia, Roberto D. [Universidade do Vale do Itajai (UNIVALI), Florianopolis, SC (Brazil). Nucleo de Investigacoes Quimico-Farmaceuticas (NIQFAR)]. E-mail: rdv@ccs.univali.br; Nascimento, Maria G.; Soldi, Valdir [Santa Catarina Univ., Florianopolis, SC (Brazil). Dept. de Quimica]. E-mail: graca@qmc.ufsc.br; vsoldi@qmc.ufsc.br

    2001-07-01

    Various lipases were immobilized in PSS/PEO blends and used as bio catalysts in the esterification reaction of lauric acid with n-pentanol, in hexane as a solvent for 24 h at 35 deg C. The best results in the ester conversion, were obtained by using lipase from Rhryzopus oryzae immobilized in PSS/PEO 80:20 blend. The data are in agreement with DSC and TGA values, which showed that these systems (blend/lipase) were very stable with low mass loss. No product was obtained by using lipase FAP-15 immobilized in PSS film , showing the strong influence of the polymer on enzyme activity. (author)

  17. A Bioactivity-Based Method for Screening, Identification of Lipase Inhibitors, and Clarifying the Effects of Processing Time on Lipase Inhibitory Activity of Polygonum Multiflorum

    Directory of Open Access Journals (Sweden)

    Yan-xu Chang

    2016-01-01

    Full Text Available Traditional Chinese medicine (TCM has been used for the treatment of many complex diseases. However, the bioactive components are always undefined. In this study, a bioactivity-based method was developed and validated to screen lipase inhibitors and evaluate the effects of processing on the lipase inhibitory activity of TCM by ultrahigh performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry and fraction collector (UHPLC/Q-TOF-MS-FC. The results showed that both Polygonum multiflorum and processed P. multiflorum extracts had inhibitory effect against lipase with IC50 values of 38.84 μg/mL and 190.6 μg/mL, respectively. Stilbenes, phenolic acid, flavonoids, and anthraquinones were considered to be the potential lipase inhibitors. Eleven potential lipase inhibitors were simultaneously determined by UHPLC. Principal component analysis (PCA was employed in exploring the effects of processing time on lipase inhibitory activity of P. multiflorum. Compared with conventional methods, a bioactivity-based method could quantitatively analyze lipase inhibitory activity of individual constituent and provide the total lipase inhibitory activity of the samples. The results demonstrated that the activity integrated UHPLC/Q-TOF-MS-FC method was an effective and powerful tool for screening and identifying lipase inhibitors from traditional Chinese medicines.

  18. Production and Characterization of Lipases by Two New Isolates of Aspergillus through Solid-State and Submerged Fermentation

    OpenAIRE

    Luciane Maria Colla; Ficanha, Aline M. M.; Juliana Rizzardi; Telma Elita Bertolin; Christian Oliveira Reinehr; Jorge Alberto Vieira Costa

    2015-01-01

    Due to the numerous applications of lipases in industry, there is a need to study their characteristics, because lipases obtained from different sources may present different properties. The aim of this work was to accomplish the partial characterization of lipases obtained through submerged fermentation and solid-state fermentation by two species of Aspergillus. Fungal strains were isolated from a diesel-contaminated soil and selected as good lipases producers. Lipases obtained through subme...

  19. Competition of Thermomyces lanuginosus lipase with its hydrolysis products at the oil-water interface.

    Science.gov (United States)

    Muth, Marco; Rothkötter, Stefanie; Paprosch, Steven; Schmid, Reiner P; Schnitzlein, Klaus

    2017-01-01

    Lipase-catalyzed hydrolysis of triglycerides yields glycerol and free fatty-acids, provided that the enzyme is non-regioselective. For an Sn-1,3 regioselective enzyme, such as lipase from Thermomyces lanuginosus, the final product is no longer glycerol but Sn-2 monoglyceride instead. However, surface active molecules generated by lipolysis may have a detrimental effect on the interfacial biocatalysis since it is known that low molecular weight surfactants can displace proteins from interfaces. By using drop profile analysis tensiometry, we evaluated the interfacial properties of the lipase-generated molecules and their competitive effect on the adsorption behavior of the lipase and on the proceeding lipolysis. Our results show that even at concentration ratios of 8.64×10(-4)M (Sn-2 monoglyceride) to 2.5×10(-7)M (lipase), the final interfacial pressure values are very similar as for the system containing the lipase alone (i.e. ∼26 mN/m). This is a strong indication that monoglycerides, as the most interfacially active products generated during regioselective lipolysis, are expelled from the oil-water interface by the lipase. We attribute this effect to intermolecular lipase-lipase interactions, resulting in a low desorption probability of the lipase. For low oleic acid concentrations, the interfacial tension is solely determined by the lipase, while for higher concentrations, lipase and oleic acid both contribute to the tension values. We propose a hypothesis based on the preferential interaction of oleic acid molecules with hydrophobic sites on the lipase. The pH dependence of the adsorption rate and the interfacial activity of the lipase were also investigated.

  20. Lipase-Catalyzed Aza-Michael Reaction on Acrylate Derivatives

    NARCIS (Netherlands)

    Steunenberg, P.; Sijm, M.; Zuilhof, H.; Sanders, J.P.M.; Scott, E.L.; Franssen, M.C.R.

    2013-01-01

    A methodology has been developed for an efficient and selective lipase-catalyzed aza-Michael reaction of various amines (primary and secondary) with a series of acrylates and alkylacrylates. Reaction parameters were tuned, and under the optimal conditions it was found that Pseudomonas stutzeri lipas

  1. Dual bioimprinting of Thermomyces lanuginosus lipase for synthesis of biodiesel

    Directory of Open Access Journals (Sweden)

    Joyeeta Mukherjee

    2016-06-01

    Full Text Available Use of biodiesel as an alternative to non-renewable sources of energy has become an attractive option in recent years. The enzymatic synthesis of biodiesel by transesterification of fats/oils with an alcohol is a much more sustainable route than the chemical method. However, cost effectiveness of the enzymatic route is a major barrier in its commercialization. In this work, a high activity biocatalyst design of Thermomyces lanuginosus lipase is made by dually bioimprinting it with substrate and a surfactant (which is believed to open up the lid covering the active site of the lipase during precipitation of the lipase in organic solvent. When the lipase was bioimprinted with only the surfactants, 28 U of the enzyme/g of oil could yield 99% biodiesel from soybean oil in about 4 h. However, when dually bioimprinted even very low enzyme load 1.4 U/g of oil, yielded 99% biodiesel within 48 h.

  2. [Cytophysiology of Penicillium solitum: a producer of lipase].

    Science.gov (United States)

    Toskueva, E P; Araviĭskiĭ, R A; Efimova, T P

    1988-09-01

    The cell population of Penicillin solitum was studied during maximum accumulation of lipase in the medium with electron microscopic and immunofluorescence methods. The data provided a conclusion that 2 types of lypolytic enzymes with various substrate and antigenic characteristics formed in the cells of P. solitum. It is likely that there is a specific inductor for exolipase synthesis as well as relationship endoenzymatic systems.

  3. Lipase catalyse glycerolysis for kinetic resolution of racemates.

    Science.gov (United States)

    Dlugy, C; Wolfson, A

    2007-09-01

    Candida antarctica lipase B catalyzed kinetic resolution of representative secondary alcohols, esters, and amine was successfully performed using triacetin or glycerol as solvents and acyl donor/acceptor. High conversions and enantioselectivities were achieved and the product was easily separated by simple extraction with diethyl ether.

  4. Triglyceride selectivity of immobilized Thermomyces lanuginosa lipase in interesterification

    DEFF Research Database (Denmark)

    Rønne, Torben Harald; Pedersen, Lars S.; Xu, Xuebing

    2005-01-01

    The triglyceride (fatty acid) selectivity of an immobilized lipase from Thermomyces lanuginosa (Lipozyme TL IM) was investigated in lipase-catalyzed interesterification reactions between two mono-acid TG in n-hexane. Tristearin (tri-C18:0) was used as a reference in a series of TG with saturated FA...... of the lipase on the different TG, indicating that Lipozyme TL IM is nonselective toward FA or TG in the system used. A response surface design was used to investigate the influence of water activities (aw) and reaction temperatures on the reactivity of Lipozyme TL IM with a system of tripalmitin (tri-C16......:0) and trilaurin (tri-C12:0) in n-hexane. An increase in temperature (40 to 60°C) was found to affect the reactivity of the lipase significantly. The reactivity of Lipozyme TL IM was unaffected by the change in aw from 0.1130 to 0.5289. An increase in aw only led to an increase in FFA formation....

  5. Lipase-catalyzed hydrolysis of methyl-3-phenylglycidate

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    The enzymatic resolution of racemic methyl 3-phenylglycidate was investigated. It was found that the hydrolysis rate of (2S, 3R)-enantiomer was faster than that of (2R, 3S)-enantiomer by a new lipase. At optimal condition 96% of (2R, 3S)-methyl phenylglycidate with ee of 100% was recovered from the racemic mixture.

  6. Microplate Bioassay for Determining Substrate Selectivity of "Candida rugosa" Lipase

    Science.gov (United States)

    Wang, Shi-zhen; Fang, Bai-shan

    2012-01-01

    Substrate selectivity of "Candida rugosa" lipase was tested using "p"-nitrophenyl esters of increasing chain length (C[subscript 1], C[subscript 7], C[subscript 15]) using the high-throughput screening method. A fast and easy 96-well microplate bioassay was developed to help students learn and practice biotechnological specificity screen. The…

  7. Specificity of Mucor miehei lipase on methyl ester substrates

    Directory of Open Access Journals (Sweden)

    Pina, M.

    1993-12-01

    Full Text Available Fatty acid methyl esters constitute a good substrate for the characterization of lipase typospecificity. In the present work, the hydrolytic action of lipase from Mucor miehei was studied. It was demonstrated that this lipase preferentially catalyses the hydrolysis of fatty acid methyl esters with small number of double bonds. It was also found that this lipase shows a specificity in the hydrolysis of fatty acid methyl esters with short aliphatic chain.Los esteres metílicos de ácidos grasos constituyen un buen sustrato para la caracterización de tipos de especificidad de lipasa. En el presente trabajo, se estudió la acción hidrolítica de lipasa de Mucor miehei. Se demostró que esta lipasa cataliza preferencialmente la hidrólisis de esteres metílicos de ácidos grasos con número pequeño de dobles enlaces. Se encontró también que esta lipasa muestra una especificidad en la hidrólisis de ásteres metílicos de ácidos grasos con cadena alifática corta.

  8. Study of microwave effects on the lipase-catalyzed hydrolysis.

    Science.gov (United States)

    Chen, Chia-Chen; Reddy, P Muralidhar; Devi, C Shobha; Chang, Po-Chi; Ho, Yen-Peng

    2016-01-01

    The effect of microwave heating on lipase-catalyzed reaction remains controversial. It is not clear whether the reaction rate enhancements are purely due to thermal/heating effects or to non-thermal effects. Therefore, quantitative mass spectrometry was used to conduct accurate kinetic analysis of lipase-catalyzed hydrolysis of triolein by microwave and conventional heating. Commercial lipases from Candida rugosa (CRL), Porcine Pancreas (PPL), and Burkholderia cepacia (BCL) were used. Hydrolysis reactions were performed at various temperatures and pH levels, along with various amounts of buffer and enzymes. Hydrolysis product yields at each time point using an internal-standard method showed no significant difference between microwave and conventional heating conditions when the reaction was carried out at the same temperature. CRL showed optimum catalytic activity at 37 °C, while PPL and BCL had better activities at 50 °C. The phosphate buffer was found to give a better hydrolysis yield than the Tris-HCl buffer. Overall results prove that a non-thermal effect does not exist in microwave-assisted lipase hydrolysis of triolein. Therefore, conventional heating at high temperatures (e.g., 50 °C) can be also used to accelerate hydrolysis reactions.

  9. Aspects of hepatic lipase expression : relation to cholesterol homeostasis

    NARCIS (Netherlands)

    D. Vieira-van Bruggen (Delfina)

    2003-01-01

    textabstractHepatic lipase has triacylglycerol hydrolase and phospholipase A1 activity towards a wide variety of substrates. It is extracellularly localized in liver and in steroid hormone producing organs. The enzyme plays an important role in both intracellular cholesterol homeostasis

  10. Lipase production by Aspergillus ibericus using olive mill wastewater.

    Science.gov (United States)

    Abrunhosa, Luís; Oliveira, Felisbela; Dantas, Danielle; Gonçalves, Cristiana; Belo, Isabel

    2013-03-01

    Olive mill wastewater (OMW) characteristics make it a suitable resource to be used as a microbial culture media to produce value-added compounds, such as enzymes. In this work, the ability of the novel species Aspergillus ibericus to discolor OMW and produce lipase was studied. An initial screening on plates containing an OMW-based agar medium and an emulsified olive oil/rhodamine-B agar medium was employed to select the strain A. ibericus MUM 03.49. Then, experiments in conical flasks with liquid OMW-based media showed that the fungus could growth on undiluted OMW, with a chemical oxygen demand (COD) of 97 ± 2 g/L, and to produce up to 2,927 ± 54 U/L of lipase. When pure OMW was used in the media, the maximum COD and color reduction achieved were 45 and 97 %, respectively. When OMW diluted to 10 % was used, A. ibericus was able to reduce phenolic and aromatic compounds by 37 and 39 %, respectively. Additionally, lipase production was found to be promoted by the addition of mineral nutrients. When the fermentations were scaled up to a 2-L bioreactor, A. ibericus produced up to 8,319 ± 33 U/L of lipase, and the maximum COD and color reduction were 57 and 24 %, respectively.

  11. Aktivitas enzim lipase alkali dari bakteri dalam surfaktan

    Directory of Open Access Journals (Sweden)

    V. Sri Pertiwi Rumiyati

    1999-07-01

    Full Text Available Activity and stability of alkaline lipase from bacteria (strain AS, KB and SP were studied in containing of surfactants at 0,05% and 0,10%. The survactanswere used in research, these were consisted of four anionic surfactants (cetyl trimethyl ammonium bromide, cetyl pyridium chloride, cetyl dimethyl ammonium bromide and there nonionic surfactants (Triton X – 100, tergitol, and nonidet P.40. Production of enzyme was produced in initial medium pH 7,5; incubation at 37oC for 48 h. These research showed that activity of alkaline lipase from strain A S-1; AS-2; KB-4; SP-2 and SP-13 was stable in containing of anionic surfactants and nonionic surfactants at 0,05% and alkaline lipase from KB-8 and SP-1 were showed decrease of activity (relatif activity < 80 %. Strain KB – 4 was prodused alkaline lipase which stable in containing of anionic surfactants, cationic surfactants and nonionic surfactants at 0,05% & 0,10%. It was had high activity (activity relatif 90-125%.

  12. Synthesis of Triptorelin Lactate Catalyzed by Lipase in Organic Media

    Science.gov (United States)

    Zhuang, Hong; Wang, Zhi; Wang, Jiaxin; Zhang, Hong; Xun, Erna; Chen, Ge; Yue, Hong; Tang, Ning; Wang, Lei

    2012-01-01

    Triptorelin lactate was successfully synthesized by porcine pancreatic lipase (PPL) in organic solvents. The effects of acyl donor, substrate ratio, organic solvent, temperature, and water activity were investigated. Under the optimum conditions, a yield of 30% for its ester could be achieved in the reaction for about 48 h. PMID:22949842

  13. Studies on the incorporation of lipase in synthetic polymerisable vesicles.

    NARCIS (Netherlands)

    Mosmuller, E.W.J.

    1993-01-01

    This thesis describes studies on the suitability of synthetic polymerisable vesicles for the incorporation and stabilisation of lipase for the bioconversion of organic chemical compounds.In chapter 1 , some characteristics are reviewed of hydrolytic enzymes, and more specific those

  14. Production and properties of lipase of Aeromonas sobria.

    Science.gov (United States)

    Takahashi, Eizo; Ito, Hidenobu; Kobayashi, Hidetomo; Yamanaka, Hiroyasu; Takeda, Yoshifumi; Balakrish Nair, Gopinath; Arimoto, Sakae; Negishi, Tomoe; Okamoto, Keinosuke

    2012-05-01

    Aeromonas have been isolated from a wide variety of aquatic environments. However the number of Aeromonas in sea water is extremely small compared to that in fresh water. In in vitro culture, Aeromonas can grow in mediums containing NaCl at a concentration of 3.0%, this concentration corresponding to that of sea water. It is unclear why the number of Aeromonas is low in sea water. Exoproteins of bacteria are thought to be important for bacterial growth and survival in the environment. Previously, the present authors have shown that mediums containing 3.0% NaCl suppress production of two proteases, serine protease and metalloprotease. In this experiment, other exoproteins whose production is influenced by the amount of NaCl in the medium were analyzed. A protein whose production is repressed in medium containing 3.0% NaCl was found and purified. Biological assay of the purified protein showed that it degrades tributyrin and hydrolyzes para-nitrophenyl-fatty acylesters. These results show that the protein is a lipase. Subsequently, the nucleotide sequence of the gene encoding the lipase was determined and the amount of mRNA of the lipase gene in the cells measured. It was found that transcription of the gene is not inhibited by NaCl in the medium. This result indicates that the lipase might be synthesized, but the folding process to become an active structure does not progress smoothly in a medium containing 3.0% NaCl.

  15. Synthesis of Triptorelin Lactate Catalyzed by Lipase in Organic Media

    Directory of Open Access Journals (Sweden)

    Hong Zhuang

    2012-08-01

    Full Text Available Triptorelin lactate was successfully synthesized by porcine pancreatic lipase (PPL in organic solvents. The effects of acyl donor, substrate ratio, organic solvent, temperature, and water activity were investigated. Under the optimum conditions, a yield of 30% for its ester could be achieved in the reaction for about 48 h.

  16. Microplate Bioassay for Determining Substrate Selectivity of "Candida rugosa" Lipase

    Science.gov (United States)

    Wang, Shi-zhen; Fang, Bai-shan

    2012-01-01

    Substrate selectivity of "Candida rugosa" lipase was tested using "p"-nitrophenyl esters of increasing chain length (C[subscript 1], C[subscript 7], C[subscript 15]) using the high-throughput screening method. A fast and easy 96-well microplate bioassay was developed to help students learn and practice biotechnological specificity screen. The…

  17. Safety evaluation of a lipase expressed in Aspergillus oryzae.

    Science.gov (United States)

    Greenough, R J; Perry, C J; Stavnsbjerg, M

    1996-02-01

    A programme of studies was conducted to establish the safety of a lipase artificially expressed in Aspergillus oryzae to be used in the detergent industry and as a processing aid in the baking industry. Laboratory animal studies were used to assess general and inhalation toxicity, skin sensitization, and skin and eye irritation. Its potential to cause mutagenicity and chromosomal aberrations was assessed in microbial and tissue culture in vitro studies. The pathogenicity of A. oryzae, the organism used to produce the lipase, was also assessed in laboratory animals. Basic ecotoxicity in a variety of test species was studied. General and inhalation toxicity was low. There was evidence of mild skin irritation. There was no evidence of eye irritation, skin sensitization, mutagenic potential, chromosomal aberrations, exotoxicity or notable pathogenicity. Comparison of these results with human exposure levels and previously published data indicates that the lipase appears safe for consumers in the given applications, requires no special occupational health precautions in manufacture and is of low environmental impact. Furthermore, the organism used in production of the lipase hs no notable pathogenicity.

  18. Production and Immobilization of Partially Purified Lipase From Penicillium chrysogenum

    Directory of Open Access Journals (Sweden)

    Shafei, M. S.

    2010-01-01

    Full Text Available An extracellular lipase from Penicillium chrysogenum produced maximal activity 225 U/mL after four days at pH 6.5. It was partially purified 4.1 fold by ammonium sulphate precipitation (70%. The enzyme was immobilized on various carriers viz. alginate, k-carrageenan and polyacrylamide gel. The immobilization yield of enzyme immobilized in kcarrageenan and polyacrylamide gel (63.41% and 48.93% respectively was low in comparison to that immobilized with alginate (81.57%. Different concentrations of alginate were tried to study their effect on lipase production. Maximum immobilization yield was observed with 3% alginate. The optimal pH of the partially purified lipase was 7.5 and the optimum temperature was 35 °C. At 60 °C the immobilized enzyme retained 62.79% of its activity. Broader pH tolerance and higher heat stability could be achieved by this method. Immobilized lipase retained 72.09% relative activity after six hydrolysis cycles.

  19. 21 CFR 862.1465 - Lipase test system.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Lipase test system. 862.1465 Section 862.1465 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES CLINICAL CHEMISTRY AND CLINICAL TOXICOLOGY DEVICES Clinical Chemistry Test Systems §...

  20. Lipase biofilm deposited by Matrix Assisted Pulsed Laser Evaporation technique

    Energy Technology Data Exchange (ETDEWEB)

    Aronne, Antonio [Department of Chemical Engineering, Materials and Industrial Production, University of Naples “Federico II”, Napoli (Italy); Bloisi, Francesco, E-mail: bloisi@na.infn.it [SPIN – CNR, Naples (Italy); Department of Physics, University of Naples “Federico II”, Napoli (Italy); Calabria, Raffaela; Califano, Valeria [Istituto Motori – CNR, Naples (Italy); Depero, Laura E. [Department of Mechanical and Industrial Engineering, University of Brescia, Brescia (Italy); Fanelli, Esther [Department of Chemical Engineering, Materials and Industrial Production, University of Naples “Federico II”, Napoli (Italy); Federici, Stefania [Department of Mechanical and Industrial Engineering, University of Brescia, Brescia (Italy); Massoli, Patrizio [Istituto Motori – CNR, Naples (Italy); Vicari, Luciano R.M. [SPIN – CNR, Naples (Italy); Department of Physics, University of Naples “Federico II”, Napoli (Italy)

    2015-05-01

    Highlights: • A lipase film was deposited with Matrix Assisted Pulsed Laser Evaporation technique. • FTIR spectra show that laser irradiation do not damage lipase molecule. • Laser fluence controls the characteristics of complex structure generated by MAPLE. - Abstract: Lipase is an enzyme that finds application in biodiesel production and for detection of esters and triglycerides in biosensors. Matrix Assisted Pulsed Laser Evaporation (MAPLE), a technique derived from Pulsed Laser Deposition (PLD) for deposition of undamaged biomolecules or polymers, is characterized by the use of a frozen target obtained from a solution/suspension of the guest material (to be deposited) in a volatile matrix (solvent). The presence of the solvent avoids or at least reduces the potential damage of guest molecules by laser radiation but only the guest material reaches the substrate in an essentially solvent-free deposition. MAPLE can be used for enzymes immobilization, essential for industrial application, allowing the development of continuous processes, an easier separation of products, the reuse of the catalyst and, in some cases, enhancing enzyme properties (pH, temperature stability, etc.) and catalytic activity in non-aqueous media. Here we show that MAPLE technique can be used to deposit undamaged lipase and that the complex structure (due to droplets generated during extraction from target) of the deposited material can be controlled by changing the laser beam fluence.

  1. Pancreatic lipase and pancreatic lipase-related protein 2, but not pancreatic lipase-related protein 1, hydrolyze retinyl palmitate in physiological conditions.

    Science.gov (United States)

    Reboul, Emmanuelle; Berton, Amélie; Moussa, Myriam; Kreuzer, Corinne; Crenon, Isabelle; Borel, Patrick

    2006-01-01

    The major sources of vitamin A in the human diet are retinyl esters (mainly retinyl palmitate) and provitamin A carotenoids. It has been shown that classical pancreatic lipase (PL) is involved in the luminal hydrolysis of retinyl palmitate (RP), but it is not known whether pancreatic lipase-related proteins 1 (PLRP1) and 2 (PLRP2), two other lipases recovered in the human pancreatic juice, are also involved. The aim of this study was to assess whether RP acts a substrate for these lipase-related proteins. Pure horse PL, horse PLRP2 and dog PLRP1 were incubated with RP solubilized in its physiological vehicles, i.e., triglyceride-rich lipid droplets, mixed micelles and vesicles. High performance liquid chromatography (HPLC) was used to assess RP hydrolysis by the free retinol released in the incubation medium. Incubation of RP-containing emulsions with horse PL and colipase resulted in RP hydrolysis (0.051+/-0.01 micromol/min/mg). This hydrolysis was abolished when colipase was not added to the medium. PLRP2 and PLRP1 were unable to hydrolyze RP solubilized in emulsions, regardless of whether colipase was added to the medium. PL hydrolyzed RP solubilized in mixed micelles as well (0.074+/-0.014 micromol/min/mg). Again, this hydrolysis was abolished in the absence of colipase. PLRP2 hydrolyzed RP solubilized in micelles but less efficiently than PL (0.023+/-0.005 micromol/min/mg). Colipase had no effect on this hydrolysis. PLRP1 was unable to hydrolyze RP solubilized in micelles, regardless of whether colipase was present or absent. Both PL and PLRP2 hydrolyzed RP solubilized in a vesicle rich-solution, and a synergic phenomenon between the two lipases was enlighten. Taken together, these results show that (1) PL hydrolyzes RP whether RP is solubilized in emulsions or in mixed micelles, (2) PLRP2 hydrolyzes RP only when RP is solubilized in mixed micelles, and (3) PLRP1 is unable to hydrolyze RP regardless of whether RP is solubilized in emulsions or in mixed

  2. Amine-functionalized magnetic nanoparticles as robust support for immobilization of Lipase

    Indian Academy of Sciences (India)

    BANALATA SAHOO; SUJAN DUTTA; DIBAKAR DHARA

    2016-07-01

    Preparation of magnetic nanoparticles with controlled size and shape along with modulation of their surface properties via introduction of functional groups holds great prospect in the field of nanotechnology. Superparamagnetic, aqueous dispersible iron oxide nanoparticles (Fe₃O₃) with amine-functionalized surface were prepared through solvothermal method, using poly(ethylene imine) (PEI), ethanolamine (EA), and 2,2' -(ethylenedioxy) bis (ethylamine) (EDBE) as amine precursors. These aminated nanoparticles were used as support for the immobilization of lipase, an important industrial enzyme. Lipase was immobilized via glutaraldehyde coupling agent. These functionalized nanoparticles were characterized by XRD, FTIR, TEM, FESEM and VSM analysis. The maximum activity was obtained for the lipase immobilized on EDBE modified Fe3O4 nanoparticles. The lipase immobilized on EDBE-Fe₃O₃ depicted 83.9% relative activity with respect to the same amount of free lipase. Moreover, lipase immobilized on EDBE-Fe₃O₃ nanoparticles demonstrated good thermal and storage stability, and easy reusability. The kinetic parameters of lipase immobilized on EDBE-Fe₃O₃ were compared with those of free lipase and the apparent Michaelis-Menten constant ofimmobilized lipase was found to be nearly same as that of free lipase.

  3. KARAKTERISASI SIFAT-SIFAT BIOKIMIA EKSTRAK KASAR LIPASE EKSTRASELULER BAKTERI Azospirillum sp.PRD1

    Directory of Open Access Journals (Sweden)

    Santi Nur Handayani

    2011-11-01

    Full Text Available Enzim lipase mempunyai peranan penting dalam katalis berbagai reaksi industri satu diantaranya pembuatan flavor melalui reaksi esterifikasi. Lipase adalah biokatalis yang berperan besar dalam aplikasi bioteknologi, seperti dalam sintesis biopolimer, biodiesel, produksi obat, dan produksi flavor. Peningkatan penggunaan lipase untuk industri mendorong dilakukan penelitian untuk mendapatkan sumber-sumber lipase baru. Sumber lipase yang potensial salah satunya adalah bakteri Azospirillum sp.PRD1 dari isolat lokal Laboratorium Mikrobiologi, Fakultas Biologi Universitas Jenderal Soedirman. Tujuan penelitian adalah untuk mendapatkan ekstrak kasar lipase dan menentukan karakteristik sifat-sifat biokimiawinya. Metode yang digunakan antara lain peremajaan bakteri Azospirillum sp.PRD1, dan produksi inokulum, penentuan waktu produksi optimum dan fase pertumbuhan bakteri, ekstraksi dan produksi ekstrak kasar lipase dan penentuan karakteristik sifat-sifat biokimiawinya. Hasil penelitian diperoleh ekstrak kasar lipase dari inokulum berumur 7 jam dan medium produksi dengan induser minyak zaitun yang diinkubasi selama 3 jam memiliki aktivitas spesifik 7,0547 Unit/mg. Lipase ekstrak kasar optimum pada pH 7, suhu 40 oC dan waktu inkubasi selama 25 menit. Lipase merupakan metaloenzim dengan kofaktor Zn2+ , Mn2+, Hg2+, Ca2+, Co2+ and Mg2+.

  4. Immobilized Pseudomonas cepacia lipase for biodiesel fuel production from soybean oil

    Energy Technology Data Exchange (ETDEWEB)

    Noureddini, H.; Gao, X.; Philkana, R.S. [Nebraska-Lincoln Univ., NE (United States). Dept. of Chemical Engineering

    2005-05-01

    Enzymatic transesterification of soybean oil with methanol and ethanol was studied. Of the nine lipases that were tested in the initial screening, lipase PS from Pseudomonas cepacia resulted in the highest yield of alkyl esters. Lipase from Pseudomonas cepacia was further investigated in immobilized form within a chemically inert, hydrophobic sol-gel support. The gel-entrapped lipase was prepared by polycondensation of hydrolyzed tetramethoxysilane and iso-butyltrimethoxysilane. Using the immobilized lipase PS, the effects of water and alcohol concentration, enzyme loading, enzyme thermal stability, and temperature in the transesterification reaction were investigated. The optimal conditions for processing 10 g of soybean oil were: 35 {sup o}C, 1:7.5 oil/methanol molar ratio, 0.5 g water and 475 mg lipase for the reactions with methanol, and 35 {sup o}C, 1:15.2 oil/ethanol molar ratio, 0.3 g water, 475 mg lipase for the reactions with ethanol. Subject to the optimal conditions, methyl and ethyl esters formation of 67 and 65 mol% in 1 h of reaction were obtained for the immobilized enzyme reactions. Upon the reaction with the immobilized lipase, the triglycerides reached negligible levels after the first 30 min of the reaction and the immobilized lipase was consistently more active than the free enzyme. The immobilized lipase also proved to be stable and lost little activity when subjected to repeated uses. (author)

  5. Solvent-induced lid opening in lipases: a molecular dynamics study.

    Science.gov (United States)

    Rehm, Sascha; Trodler, Peter; Pleiss, Jürgen

    2010-11-01

    In most lipases, a mobile lid covers the substrate binding site. In this closed structure, the lipase is assumed to be inactive. Upon activation of the lipase by contact with a hydrophobic solvent or at a hydrophobic interface, the lid opens. In its open structure, the substrate binding site is accessible and the lipase is active. The molecular mechanism of this interfacial activation was studied for three lipases (from Candida rugosa, Rhizomucor miehei, and Thermomyces lanuginosa) by multiple molecular dynamics simulations for 25 ns without applying restraints or external forces. As initial structures of the simulations, the closed and open structures of the lipases were used. Both the closed and the open structure were simulated in water and in an organic solvent, toluene. In simulations of the closed lipases in water, no conformational transition was observed. However, in three independent simulations of the closed lipases in toluene the lid gradually opened. Thus, pathways of the conformational transitions were investigated and possible kinetic bottlenecks were suggested. The open structures in toluene were stable, but in water the lid of all three lipases moved towards the closed structure and partially unfolded. Thus, in all three lipases opening and closing was driven by the solvent and independent of a bound substrate molecule.

  6. Purification and characterization of an extracellular lipase from Mucor hiemalis f. corticola isolated from soil.

    Science.gov (United States)

    Ulker, Serdar; Karaoğlu, Sengül Alpay

    2012-10-01

    We have screened 39 microfungi isolates originated from soil in terms of lipolytic activity. Out of all screened, a novel strain of Mucor hiemalis f. corticola was determined to have the highest lipase activity. The extracellular lipase was produced in response to 2% glucose and 2.1% peptone. The lipase was purified 12.63-folds with a final yield of 27.7% through following purification steps; ammonium sulfate precipitation, dialysis, gel filtration column chromatography and ion exchange chromatography, respectively. MALDI-TOF MS analysis revealed 31% amino-acid identity to a known lipase from Rhizomucor miehei species. The molecular weight of the lipase was determined as 46 kDa using SDS-PAGE and analytical gel filtration. Optimal pH and temperature of the lipase were determined as 7.0 and 40°C, respectively. The enzyme activity was observed to be stable at the pH range of 7.0-9.0. Thermostability assays demonstrated that the lipase was stable up to 50°C for 60 min. The lipase was more stable in ethanol and methanol than other organic solvents tested. Furthermore, the activity of the lipase was slightly enhanced by SDS and PMSF. In the presence of p-NPP as substrate, K(m) and V(max) values of the lipase were calculated by Hanes-Woolf plot as 1.327 mM and 91.11 μmol/min, respectively.

  7. Structure of product-bound SMG1 lipase: active site gating implications.

    Science.gov (United States)

    Guo, Shaohua; Xu, Jinxin; Pavlidis, Ioannis V; Lan, Dongming; Bornscheuer, Uwe T; Liu, Jinsong; Wang, Yonghua

    2015-12-01

    Monoacylglycerol and diacylglycerol lipases are industrially interesting enzymes, due to the health benefits that arise from the consumption of diglycerides compared to the traditional triglyceride oils. Most lipases possess an α-helix (lid) directly over the catalytic pocket which regulates the activity of the enzyme. Generally, lipases exist in active and inactive conformations, depending on the positioning of this lid subdomain. However, lipase SMG1, a monoacylglycerol and diacylglycerol specific lipase, has an atypical activation mechanism. In the present study we were able to prove by crystallography, in silico analysis and activity tests that only two positions, residues 102 and 278, are responsible for a gating mechanism that regulates the active and inactive states of the lipase, and that no significant structural changes take place during activation except for oxyanion hole formation. The elucidation of the gating effect provided data enabling the rational design of improved lipases with 6-fold increase in the hydrolytic activity toward diacylglycerols, just by providing additional substrate stabilization with a single mutation (F278N or F278T). Due to the conservation of F278 among the monoacylglycerol and diacylglycerol lipases in the Rhizomucor miehei lipase-like family, the gating mechanism described herein might represent a general mechanism applicable to other monoacylglycerol and diacylglycerol lipases as well. Database: Structural data are available in the Protein Data Bank under the accession numbers 4ZRE (F278D mutant) and 4ZRD (F278N mutant).

  8. NEW LIPASE-PRODUCERS MICROORGANISMS FROM PERUVIAN AMAZONIA WHICH HYDROLYZE PALM OIL AND DERIVATIVES

    Directory of Open Access Journals (Sweden)

    Roxana Trujillo

    2014-04-01

    Full Text Available Two yeasts: Cryptococcus uchicensis TMY9 and Pichia uchicensis TMY10 and one fungus Verticillium tingalensis TMFMB are described for the first time as lipase producer microorganisms. The strains have been isolated after an ecological screening in a palm oil industry. The yeasts- C. uchicensis and Pichia uchicensis - mainly produce extracellular lipases as active as those produced by traditional lipase producing microorganisms. The extracellular lipases are active in the hydrolysis of crude palm oil and its industrial derivatives. Contrarily in the isolated fungus, the lipase mainly remains bonded to biomass. In all cases, greater hydrolytic activities are observed in the hydrolysis of palm olein and super-olein than with saturated substrates as stearine. P. uchicensis lipase shows moderated selectivity versus saturated acid triglycerides compared to substrates with high proportion of oleic acid (olein or superolein. The opposite behavior is observed with C. uchicensis and fungal lipases. P. uchicensis produces a more active crude lipase than C. uchicensis with lower biomass production. The kinetic runs performed with crude yeast lipases suggest a three steps mechanism where the high penetration of lipase in the fat gouts favors the hydrolysis.

  9. Optimization of lipase entrapment in alginate gel bead for palm olein hydrolysis

    Directory of Open Access Journals (Sweden)

    Cheirsilp, B.

    2007-05-01

    Full Text Available Lipase from Pseudomonas sp. was entrapped by drop-wise addition of an aqueous mixture of alginate and the biocatalyst to hardening solution of CaCl2 for the purpose of palm olein hydrolysis. Effects ofimmobilization conditions including alginate concentration, CaCl2 concentration, enzyme concentration and bead size on immobilized yield, immobilized lipase activity and recovery of activity (specific activity ratio ofentrapped lipase to free lipase were investigated. An increase in alginate concentration raised immobilized yield, but decreased immobilized lipase activity and recovery of activity. CaCl2 concentration in the testedrange of 50-200 mM had slight effects on immobilized yield, immobilized lipase activity and recovery of activity. In contrast to immobilized lipase activity, immobilized yield and recovery of activity decreased withincreasing enzyme concentration. With increasing bead size, immobilized lipase activity and recovery of activity decreased due to mass transfer resistance whereas immobilized yield was unchanged. The optimumcondition for lipase entrapment in alginate gel bead was alginate concentration at 2% (w/v, CaCl2 concentration at 100 mM, enzyme concentration at 30 U/ml and bead size at 2 mm. Under this entrapmentcondition, 8.11 U/ml of immobilized lipase was obtained with 95.2% of immobilized yield and 22.2% of recovery of activity.

  10. Lipase applications in oil hydrolysis with a case study on castor oil: a review.

    Science.gov (United States)

    Goswami, Debajyoti; Basu, Jayanta Kumar; De, Sirshendu

    2013-03-01

    Lipase (triacylglycerol acylhydrolase) is a unique enzyme which can catalyze various types of reactions such as hydrolysis, esterification, alcoholysis etc. In particular, hydrolysis of vegetable oil with lipase as a catalyst is widely studied. Free lipase, lipase immobilized on suitable support, lipase encapsulated in a reverse micelle and lipase immobilized on a suitable membrane to be used in membrane reactor are the most common ways of employing lipase in oil hydrolysis. Castor oil is a unique vegetable oil as it contains high amounts (90%) of a hydroxy monounsaturated fatty acid named ricinoleic acid. This industrially important acid can be obtained by hydrolysis of castor oil. Different conventional hydrolysis processes have certain disadvantages which can be avoided by a lipase-catalyzed process. The degree of hydrolysis varies widely for different lipases depending on the operating range of process variables such as temperature, pH and enzyme loading. Immobilization of lipase on a suitable support can enhance hydrolysis by suppressing thermal inactivation and estolide formation. The presence of metal ions also affects lipase-catalyzed hydrolysis of castor oil. Even a particular ion has different effects on the activity of different lipases. Hydrophobic organic solvents perform better than hydrophilic solvents during the reaction. Sonication considerably increases hydrolysis in case of lipolase. The effects of additives on the same lipase vary with their types. Nonionic surfactants enhance hydrolysis whereas cationic and anionic surfactants decrease it. A single variable optimization method is used to obtain optimum conditions. In order to eliminate its disadvantages, a statistical optimization method is used in recent studies. Statistical optimization shows that interactions between any two of the following pH, enzyme concentration and buffer concentration become significant in presence of a nonionic surfactant named Span 80.

  11. Post-heparin plasma lipoprotein lipase, but not hepatic lipase activity, is related to plasma adiponectin in type 2 diabetic patients and healthy subjects

    NARCIS (Netherlands)

    De Vries, R; Wolffenbuttel, BHR; Sluiter, WJ; Van Tol, A; Dullaart, RPF

    2005-01-01

    The aim of this study was to determine the relationships of plasma adiponectin with post-heparin plasma lipoprotein lipase (LPL) and hepatic lipase (HL) activities, and to evaluate whether plasma adiponectin contributes to diabetes-associated dyslipidaemia. Plasma adiponectin, post-heparin plasma

  12. The G-250A polymorphism in the hepatic lipase gene promoter is associated with changes in hepatic lipase activity and LDL cholesterol: The KANWU Study

    DEFF Research Database (Denmark)

    Lindi, Virpi; Schwab, Ursula; Louheranta, Anne;

    2007-01-01

    BACKGROUND AND AIMS: Hepatic lipase (HL) catalyzes the hydrolysis of triglycerides and phospholipids from lipoproteins, and promotes the hepatic uptake of lipoproteins. A common G-250A polymorphism in the promoter of the hepatic lipase gene (LIPC) has been described. The aim was to study...

  13. Lipase catalyzed ester synthesis for food processing industries

    Directory of Open Access Journals (Sweden)

    Aravindan Rajendran

    2009-02-01

    Full Text Available Lipases are one of the most important industrial biocatalyst which catalyzes the hydrolysis of lipids. It can also reverse the reaction at minimum water activity. Because of this pliable nature, it is widely exploited to catalyze the diverse bioconversion reactions, such as hydrolysis, esterification, interesterification, alcoholysis, acidolysis and aminolysis. The property to synthesize the esters from the fatty acids and glycerol promotes its use in various ester synthesis. The esters synthesized by lipase finds applications in numerous fields such as biodiesel production, resolution of the recemic drugs, fat and lipid modification, flavour synthesis, synthesis of enantiopure pharmaceuticals and nutraceuticals. It plays a crucial role in the food processing industries since the process is unaffected by the unwanted side products. Lipase modifications such as the surfactant coating, molecular imprinting to suit for the non-aqueous ester synthesis have also been reported. This review deals with lipase catalyzed ester synthesis, esterification strategies, optimum conditions and their applications in food processing industries.Lipases são catalizadores industriais dos mais importantes, os quais catalizam a hidrólise de lipídeos. Também podem reverter a reação a um mínimo de atividade de água. Devido sua natureza flexível, é amplamente explorada para catalizar uma diversidade de reações de bioconversão como hidrólise, esterificação, interesterificação, alcoólise, acidólise e aminólise. A propriedade de síntese de esteres a partir de ácidos graxos e glicerol promoveu seu uso em várias sínteses de esteres. Os esteres sintetizados por lipases encontram aplicação em numerosos campos como a produção de biodiesel, resolução de drogas racêmicas, modificação de gorduras e lipídios, sintese de aromas, síntese de produtos farmacêuticos enantiopuro e nutracêuticos. As lipases possuem um papel crucial nas indústrias de

  14. Analysis of Comparative Sequence and Genomic Data to Verify Phylogenetic Relationship and Explore a New Subfamily of Bacterial Lipases.

    Directory of Open Access Journals (Sweden)

    Malihe Masomian

    Full Text Available Thermostable and organic solvent-tolerant enzymes have significant potential in a wide range of synthetic reactions in industry due to their inherent stability at high temperatures and their ability to endure harsh organic solvents. In this study, a novel gene encoding a true lipase was isolated by construction of a genomic DNA library of thermophilic Aneurinibacillus thermoaerophilus strain HZ into Escherichia coli plasmid vector. Sequence analysis revealed that HZ lipase had 62% identity to putative lipase from Bacillus pseudomycoides. The closely characterized lipases to the HZ lipase gene are from thermostable Bacillus and Geobacillus lipases belonging to the subfamily I.5 with ≤ 57% identity. The amino acid sequence analysis of HZ lipase determined a conserved pentapeptide containing the active serine, GHSMG and a Ca(2+-binding motif, GCYGSD in the enzyme. Protein structure modeling showed that HZ lipase consisted of an α/β hydrolase fold and a lid domain. Protein sequence alignment, conserved regions analysis, clustal distance matrix and amino acid composition illustrated differences between HZ lipase and other thermostable lipases. Phylogenetic analysis revealed that this lipase represented a new subfamily of family I of bacterial true lipases, classified as family I.9. The HZ lipase was expressed under promoter Plac using IPTG and was characterized. The recombinant enzyme showed optimal activity at 65 °C and retained ≥ 97% activity after incubation at 50 °C for 1h. The HZ lipase was stable in various polar and non-polar organic solvents.

  15. Cloning and expression of gene, and activation of an organic solvent-stable lipase from Pseudomonas aeruginosa LST-03.

    Science.gov (United States)

    Ogino, Hiroyasu; Katou, Yoshikazu; Akagi, Rieko; Mimitsuka, Takashi; Hiroshima, Shinichi; Gemba, Yuichi; Doukyu, Noriyuki; Yasuda, Masahiro; Ishimi, Kosaku; Ishikawa, Haruo

    2007-11-01

    Organic solvent-tolerant Pseudomonas aeruginosa LST-03 secretes an organic solvent-stable lipase, LST-03 lipase. The gene of the LST-03 lipase (Lip9) and the gene of the lipase-specific foldase (Lif9) were cloned and expressed in Escherichia coli. In the cloned 2.6 kbps DNA fragment, two open reading frames, Lip9 consisting of 933 nucleotides which encoded 311 amino acids and Lif9 consisting of 1,020 nucleotides which encoded 340 amino acids, were found. The overexpression of the lipase gene (lip9) was achieved when T7 promoter was used and the signal peptide of the lipase was deleted. The expressed amount of the lipase was greatly increased and overexpressed lipase formed inclusion body in E. coli cell. The collected inclusion body of the lipase from the cell was easily solubilized by urea and activated by using lipase-specific foldase of which 52 or 58 amino acids of N-terminal were deleted. Especially, the N-terminal methionine of the lipase of which the signal peptide was deleted was released in E. coli and the amino acid sequence was in agreement with that of the originally-produced lipase by P. aeruginosa LST-03. Furthermore, the overexpressed and solubilized lipase of which the signal peptide was deleted was more effectively activated by lipase-specific foldase.

  16. HIDROLISIS ENZIMATIS STEARIN SAWIT MENJADI MONOGLISERIDA OLEH LIPASE DARI Rhizomucor miehei DAN PANKREAS (Enzymatic Hydrolysis of Palm Stearin to Produce Monoglyceride by Lipase from Rhizomucor miehei and Pancreatic

    Directory of Open Access Journals (Sweden)

    Steivie Karaouw

    2013-06-01

    Full Text Available The objectives of the research were to evaluate the effect of the pH, ratio of substrate:phospate buffer, and reaction time on the enzymatic hydrolysis of palm stearin to obtain monoglyceride by R. miehei and pancreatic lipases. Hydrolysis was evaluated at various pH (6.0; 6.5; 7.0; 7.5 dan 8.0. Enzymatic hydrolysis reactions were held at various ratio of substrate:phospate buffer (10:1, 10:2, 10:3, 10:4, 10:5, 10:6 and duration time of 6, 12, 18, 24 hours by R. miehei lipase and 24, 30, 36, 42, 48 hours by pancreatic lipase. Enzymatic hydrolysis reaction was carried out in waterbath shaker 80 stroke/minute, at 40oC with R.miehei lipase and 37oC with pancreatic lipase. The hydrolysis products were monitored using TLC with petroleum ether:diethyl ether:acetic acid=60:40:1 as developing solvent on silica gel F254 20×20 cm plate. The results showed that optimum pH for both R. miehei and pancreatic lipases were 6.5 and their activities were 332.25 unit/g enzyme amobile and 228.04 unit/g enzyme, respectively. The highest monoglyceride fraction was obtained from ratio substrate:phospate buffer 10:1 at 18 hours of incubation by Rhizomucor miehei lipase (21,59% and ratio substrate:phospate buffer 10:4 at 42 hours of incubation by pancreatic lipase (40,45%. Keywords: Hydrolysis, palm stearin, monoglyceride, lipase, Rhizomucor miehei, pancreatic   ABSTRAK Penelitian ini bertujuan untuk mengetahui pengaruh pH, rasio substrat:buffer fosfat dan waktu hidrolisis terhadap produksi monogliserida 2-monopalmitin secara enzimatis menggunakan lipase dari Rhizomucor miehei dan lipase pankreas. Hidrolisis dilakukan pada pH (6,0; 6,5; 7,0; 7,5 dan 8,0, dengan rasio substrat:buffer fosfat (10:1, 10:2, 10:3, 10:4, 10:5 dan 10:6 dan waktu hidrolisis (6, 12, 18 dan 24 jam menggunakan lipase dari R. miehei dan (6, 12, 18, 24, 30, 36, 42 dan 48 jam menggunakan lipase pankreas. Reaksi hidrolisis berlangsung dalam shaker waterbath 80 stroke/menit, pada suhu 40oC untuk

  17. Lipase production by yeasts from extra virgin olive oil.

    Science.gov (United States)

    Ciafardini, G; Zullo, B A; Iride, A

    2006-02-01

    Newly produced olive oil has an opalescent appearance due to the presence of solid particles and micro-drops of vegetation water from the fruits. Some of our recent microbiological research has shown that a rich micro-flora is present in the suspended fraction of the freshly produced olive oil capable of improving the quality of the oil through the hydrolysis of the oleuropein. Present research however has, for the first time, demonstrated the presence of lipase-positive yeasts in some samples of extra virgin olive oil which can lower the quality of the oil through the hydrolysis of the triglycerides. The tests performed with yeasts of our collection, previously isolated from olive oil, demonstrated that two lipase-producing yeast strains named Saccharomyces cerevisiae 1525 and Williopsis californica 1639 were able to hydrolyse different specific synthetic substrates represented by p-nitrophenyl stearate, 4-nitrophenyl palmitate, tripalmitin and triolein as well as olive oil triglycerides. The lipase activity in S. cerevisiae 1525 was confined to the whole cells, whereas in W. californica 1639 it was also detected in the extracellular fraction. The enzyme activity in both yeasts was influenced by the ratio of the aqueous to the organic phase reaching its maximum value in S. cerevisiae 1525 when the water added to the olive oil was present in a ratio of 0.25% (v/v), whereas in W. californica 1639 the optimal ratio was 1% (v/v). Furthermore, the free fatty acids of olive oil proved to be good inducers of lipase activity in both yeasts. The microbiological analysis carried out on commercial extra virgin olive oil, produced in four different geographic areas, demonstrated that the presence of lipase-producing yeast varied from zero to 56% of the total yeasts detected, according to the source of oil samples. The discovery of lipase-positive yeasts in some extra virgin olive oils leads us to believe that yeasts are able to contribute in a positive or negative way towards

  18. Covalent immobilization of lipases on monodisperse magnetic microspheres modified with PAMAM-dendrimer

    Energy Technology Data Exchange (ETDEWEB)

    Zhu, Weiwei [Lanzhou University, State Key Laboratory of Applied Organic Chemistry, Key Laboratory of Nonferrous Metal Chemistry and Resources Utilization of Gansu Province, College of Chemistry and Chemical Engineering, Institute of Biochemical Engineering and Environmental Technology (China); Zhang, Yimei [Suzhou Research Academy of North China Electric Power University (China); Hou, Chen; Pan, Duo; He, Jianjun; Zhu, Hao, E-mail: zhuhao07@lzu.edu.cn [Lanzhou University, State Key Laboratory of Applied Organic Chemistry, Key Laboratory of Nonferrous Metal Chemistry and Resources Utilization of Gansu Province, College of Chemistry and Chemical Engineering, Institute of Biochemical Engineering and Environmental Technology (China)

    2016-02-15

    This paper reported an immobilization of Candida rugosa lipase (CRL) onto PAMAM-dendrimer-grafted magnetic nanoparticles synthesized by a modified solvothermal reduction method. The dendritic magnetic nanoparticles were amply characterized by several instrumental measurements, and the CRL was covalently anchored on the three generation supports with glutaraldehyde as coupling reagent. The amount of immobilized enzyme was up to 150 mg/g support and the factors related with the enzyme activity were investigated. The immobilization of lipase improved their performance in wider ranges of pH and temperature. The immobilized lipase exhibited excellent thermal stability and reusability in comparison with free enzyme and can be reused 10 cycles with the enzymatic activity remained above 90 %. The properties of lipase improved obviously after being immobilized on the dendritic supports. The inactive immobilized lipase could be regenerated with glutaraldehyde and Cu{sup 2+}, respectively. This synthetic strategy was facile and eco-friendly for applications in lipase immobilization.

  19. Characterization of Lipoprotein Lipases interactions with Sortilin and SorLA

    DEFF Research Database (Denmark)

    Klinger, Stine Christensen

    . The present study describes their trafficking of two ligands, lipoprotein lipase and apolipoprotein A-V, which are integral parts of the lipid metabolism. Lipoprotein lipase is found at the luminal side of capillary endothelial cells, where it is involved in the conversion of circulating triglycerides to free...... fatty acids, which in turn are used as an energy source in muscle cells or as an energy reserve in adipose tissue. Lack of lipoprotein lipase leads to an elevated level of plasma lipids and results in increased risk of cardiovascular diseases. The regulation of lipoprotein lipase expression takes place...... at both transcriptional and posttranslational levels. While the transcriptional regulation of the lipase is well described, the posttranslational mechanisms affecting lipoprotein lipase expression are far from understood. The functions of the recently discovered apolipoprotein A-V are still a subject...

  20. Conversion of sunflower oil to biodiesel by alcoholysis using immobilized lipase.

    Science.gov (United States)

    Sagiroglu, Ayten

    2008-01-01

    Transesterification reaction was performed using sunflower oil and short-chain alcohol by immobilized lipases in organic solvents. The fatty acid ester, which is the product of this reaction, can be used as a diesel fuel that does not produce sulfur oxide and minimize the soot particulate. Immobilized porcine pancreatic lipase (PPL) and Candida rugosa lipase (CRL) showed the satisfactory activity in these reactions. Immobilization of lipases was carried out using inorganic absorbance Celit 545 particle as a carrier. Organic solvent like hexane in reactions was required when methanol and ethanol were used as alcoholic substrate. The reaction could be performed in absence of solvent when 1-propanol and 1-butanol were used as short-chain alcohol. The activities of immobilized lipases were highly increased in comparison with free lipases because its activity sites became more effective. Immobilized enzyme could be repeatedly used without difficult method of separation and the decrease in its activity was not largely observed.

  1. Biodiesel production from Jatropha oil catalyzed by immobilized Burkholderia cepacia lipase on modified attapulgite.

    Science.gov (United States)

    You, Qinghong; Yin, Xiulian; Zhao, Yuping; Zhang, Yan

    2013-11-01

    Lipase from Burkholderia cepacia was immobilized on modified attapulgite by cross-linking reaction for biodiesel production with jatropha oil as feedstock. Effects of various factors on biodiesel production were studied by single-factor experiment. Results indicated that the best conditions for biodiesel preparation were: 10 g jatropha oil, 2.4 g methanol (molar ratio of oil to methanol is 1:6.6) being added at 3h intervals, 7 wt% water, 10 wt% immobilized lipase, temperature 35°C, and time 24h. Under these conditions, the maximum biodiesel yield reached 94%. The immobilized lipase retained 95% of its relative activity during the ten repeated batch reactions. The half-life time of the immobilized lipase is 731 h. Kinetics was studied and the Vmax of the immobilized lipases were 6.823 mmol L(-1). This immobilized lipase catalyzed process has potential industrial use for biodiesel production to replace chemical-catalyzed method.

  2. Human pancreatic triglyceride lipase expressed in yeast cells: purification and characterization.

    Science.gov (United States)

    Yang, Y; Lowe, M E

    1998-06-01

    A cDNA clone encoding human pancreatic triglyceride lipase was cloned into a yeast expression vector so that the yeast PHO1 signal peptide replaced the native signal peptide. Pichia pastoris cells were transfected with the vector, and clones expressing human pancreatic triglyceride lipase were isolated. Recombinant human pancreatic lipase was expressed in broth cultures and was purified from the medium by DEAE blue Sepharose and hydroxyapatite chromatography. The highly purified lipase had specific activities for various triglyceride substrates identical to those of tissue-purified human pancreatic triglyceride lipase; it was inhibited by bile salts, required colipase for activity, and demonstrated interfacial activation. This expression system is suitable for the rapid, efficient production of human pancreatic triglyceride lipase in amounts adequate for biophysical studies.

  3. Functional expression of Rhizopus oryzae lipase in Pichia pastoris: high-level production and some properties.

    Science.gov (United States)

    Minning, S; Schmidt-Dannert, C; Schmid, R D

    1998-12-11

    The mature lipase of the fungus Rhizopus oryzae (ROL) was functionally expressed and secreted in the methylotrophic yeast Pichia pastoris. In a batch cultivation, where methanol feeding was linked to the dissolved oxygen content in the cultivation solution, a lipase activity of 500,000 units per liter (60 mg active lipase per liter) of culture was achieved after initial glycerol feeding of the culture. Recombinant ROL lipase was purified to homogeneity by a simple two-step purification procedure and had a specific activity of 8571 U mg-1 (triolein, 30 degrees C, pH 8.1) which is comparable with the purified native enzyme. The properties of the recombinant lipase were similar to those reported both for the native lipase and for the enzyme expressed in Escherichia coli and refolded from inactive inclusion bodies.

  4. Collagen-Immobilized Lipases Show Good Activity and Reusability for Butyl Butyrate Synthesis.

    Science.gov (United States)

    Dewei, Song; Min, Chen; Haiming, Cheng

    2016-11-01

    Candida rugosa lipases were immobilized onto collagen fibers through glutaraldehyde cross-linking method. The immobilization process has been optimized. Under the optimal immobilization conditions, the activity of the collagen-immobilized lipase reached 340 U/g. The activity was recovered of 28.3 % by immobilization. The operational stability of the obtained collagen-immobilized lipase for hydrolysis of olive oil emulsion was determined. The collagen-immobilized lipase showed good tolerance to temperature and pH variations in comparison to free lipase. The collagen-immobilized lipase was also applied as biocatalyst for synthesis of butyl butyrate from butyric acid and 1-butanol in n-hexane. The conversion yield was 94 % at the optimal conditions. Of its initial activity, 64 % was retained after 5 cycles for synthesizing butyl butyrate in n-hexane.

  5. Determination of lipase activity in the larval midgut of Bacterocera oleae Gmelin (Diptera: Tephritidae

    Directory of Open Access Journals (Sweden)

    S Delkash-Roudsari

    2014-02-01

    Full Text Available In the current study, digestive lipase activity was determined and characterized in the third larval instars of olive fly, Bactericera oleae as the first time in dipteran order. By using two sample fractions, it was found that the enzyme had higher activity in membrane-bound fraction than that of soluble fraction. Optimal pH of soluble lipase was found to be 4 and 6 but membrane-bound lipase showed pH 4 as optimal value. Optimal temperatures for soluble and membrane-bound lipase were obtained to be 35 and 50 °C, respectively. Activities of digestive soluble and membrane-bound lipases decreased by using various mono- and di-valent ions. Since, fruits of olive are full of various oils, digestive lipases of B. oleae larvae have critical role in their digestion. So, these enzymes might be a good target for developing inhibitors and resistant varieties.

  6. Partial characterization of pyloric-duodenal lipase of gilthead seabream (Sparus aurata).

    Science.gov (United States)

    Nolasco, Héctor; Moyano-López, Francisco; Vega-Villasante, Fernando

    2011-03-01

    In the present study, we report the isolation and characterization of seabream Sparus aurata pyloric caeca-duodenal lipase. Optimum activity was found at pH 8.5 and salinity of 50 mM NaCl. Lipase activity was sensitive to divalent ions, and extreme pH values (4, 5, and 12), being more stable at alkaline than acid pH. Optimum temperature was found at 50°C, but lipase was stable at temperatures below 40°C. Lipase has a bile salt sodium taurocholate requirement for increased activity. Gradient PAGE electrophoresis revealed the presence of four isoforms with apparent molecular masses of 34, 50, 68, and 84 KDa, respectively. Pyloric-duodenal lipase was able to hydrolyze emulsified alimentary oils. Results confirm the presence of true lipases in Sparus aurata digestive tract.

  7. Effects of methanol on lipases: molecular, kinetic and process issues in the production of biodiesel.

    Science.gov (United States)

    Lotti, Marina; Pleiss, Jürgen; Valero, Francisco; Ferrer, Pau

    2015-01-01

    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.

  8. Streptomyces rimosus GDS(L Lipase: Production, Heterologous Overexpression and Structure-Stability Relationship

    Directory of Open Access Journals (Sweden)

    Marija Abramić

    2003-01-01

    Full Text Available Streptomyces rimosus lipase gene has been overexpressed in a heterologous host, S. lividans TK23. The maximal lipase activity was determined in the culture filtrates of the late stationary phase. Time course of lipase production was monitored by a modified plate assay. S. rimosus lipase gene has been located on the AseI B fragment approximately 2 Mb far from the left end of the S. rimosus linear chromosome. Out of eight examined streptomycetes, the presence of this rare type of bacterial lipase gene was detected in two belonging to the S. rimosus taxonomic cluster, and in one non-related species. Comparison of protein sequences of the Streptomyces lipolytic enzymes was performed. The result indicated the best structural stability of the putative S. coelicolor lipase-2.

  9. The Crystal Structure of Bacillus subtilis Lipase : A Minimal α/β Hydrolase Fold Enzyme

    NARCIS (Netherlands)

    Pouderoyen, Gertie van; Eggert, Thorsten; Jaeger, Karl-Erich; Dijkstra, Bauke W.

    2001-01-01

    The X-ray structure of the lipase LipA from Bacillus subtilis has been determined at 1.5 Å resolution. It is the first structure of a member of homology family I.4 of bacterial lipases. The lipase shows a compact minimal α/β hydrolase fold with a six-stranded parallel β-sheet flanked by five α-helic

  10. Production and Characterization of Biodiesel Using Nonedible Castor Oil by Immobilized Lipase from Bacillus aerius

    OpenAIRE

    Sunil Kumar Narwal; Nitin Kumar Saun; Priyanka Dogra; Ghanshyam Chauhan; Reena Gupta

    2015-01-01

    A novel thermotolerant lipase from Bacillus aerius was immobilized on inexpensive silica gel matrix. The immobilized lipase was used for the synthesis of biodiesel using castor oil as a substrate in a solvent free system at 55°C under shaking in a chemical reactor. Several crucial parameters affecting biodiesel yield such as incubation time, temperature, substrate molar ratio, and amount of lipase were optimized. Under the optimized conditions, the highest biodiesel yield was up to 78.13%. Th...

  11. Beauveria bassiana Lipase A expressed in Komagataella (Pichia) pastoris with potential for biodiesel catalysis

    OpenAIRE

    Ana Claudia Vici; Andrezza Furquim da Cruz; Fernanda Dell Antonio Facchini; Caio Cesar Carvalho; Marita Giminez Pereira; Raquel eFonseca-Maldonado; Richard John Ward; Benevides Costa Pessela; Gloria eFernadez-Lorente; Fernando Araripe Gonçalves Torres; João Atílio Jorge; Maria de Lourdes Teixeira de Moraes Polizeli

    2015-01-01

    Lipases (EC 3.1.1.3) comprise a biotechnologically important group of enzymes because they are able to catalyze both hydrolysis and synthesis reactions, depending on the amount of water in the system. One of the most interesting application of lipase is in the biofuel industry for biodiesel production by oil and ethanol (or methanol) transesterification. Entomopathogenic fungi, which are potential source of lipases, are still poorly explored in biotechnological processes. The present work rep...

  12. Immobilization and catalytic properties of lipase on chitosan for hydrolysis and esterification reactions

    OpenAIRE

    2003-01-01

    The objective of this study was to evaluate the immobilization of lipase on a chitosan support by physical adsorption, aiming at its application in hydrolytic and synthetic reactions. Two types of chitosan (flakes and porous) were used for immobilizing lipase from a microbial source (Candida rugosa) and animal cells (porcine pancreas). The best results for recovery of total activity after immobilization were obtained for microbial lipase and porous chitosan beads. This set was selected for fu...

  13. Lipase-catalyzed enantioselective esterification of flurbiprofen with n-butanol

    OpenAIRE

    2000-01-01

    The influences of water activity and solvent hydrophobicity on the kinetics of the lipase-catalyzed enantioselective esterification of flurbiprofen with n-butanol were investigated. The solvent effect was not similar for lipases from Candida rugosa (Crl), Mucor javanicus (Mjl), and porcine pancreas (Ppl). The lipase-catalyzed reaction rates in different solvents across a wide range of water activities revealed that the Ppl-catalyzed reaction exhibited no enantioselectivity and no substantial ...

  14. Characterization of lipases from Staphylococcus aureus and Staphylococcus epidermidis isolated from human facial sebaceous skin.

    Science.gov (United States)

    Xie, Winny; Khosasih, Vivia; Suwanto, Antonius; Kim, Hyung Kwoun

    2012-01-01

    Two staphylococcal lipases were obtained from Staphylococcus epidermidis S2 and Staphylococcus aureus S11 isolated from sebaceous areas on the skin of the human face. The molecular mass of both enzymes was estimated to be 45 kDa by SDS-PAGE. S2 lipase displayed its highest activity in the hydrolysis of olive oil at 32 degrees C and pH 8, whereas S11 lipase showed optimal activity at 31 degrees C and pH 8.5. The S2 lipase showed the property of cold-adaptation, with activation energy of 6.52 kcal/mol. In contrast, S11 lipase's activation energy, at 21 kcal/mol, was more characteristic of mesophilic lipases. S2 lipase was stable up to 45° C and within the pH range from 5 to 9, whereas S11 lipase was stable up to 50 degrees C and from pH 6 to 10. Both enzymes had high activity against tributyrin, waste soybean oil, and fish oil. Sequence analysis of the S2 lipase gene showed an open reading frame of 2,067 bp encoding a signal peptide (35 aa), a pro-peptide (267 aa), and a mature enzyme (386 aa); the S11 lipase gene, at 2,076 bp, also encoded a signal peptide (37 aa), pro-peptide (255 aa), and mature enzyme (399 aa). The two enzymes maintained amino acid sequence identity of 98-99% with other similar staphylococcal lipases. Their microbial origins and biochemical properties may make these staphylococcal lipases isolated from facial sebaceous skin suitable for use as catalysts in the cosmetic, medicinal, food, or detergent industries.

  15. Biocatalytic Synthesis of Polyglycerol Polyricinoleate: A Comparison of Different Commercial Lipases

    OpenAIRE

    Ortega, S.; Gómez, J. L.; Bastida, J.; Máximo, M. F.; Montiel, M. C.; Gómez, M.

    2013-01-01

    This paper describes the studies carried out to select the most suitable lipase as catalyst for the esterification of polyglycerol with polyricinoleic acid to yield polyglicerol polyricinoleate (PGPR), a value-added, bio-based food emulsifier. The enzymes assayed were lipases from Rhizopus arrhizus, Rhizopus oryzae and Mucor javanicus, previously selected because of their suitable activity and moderate cost. First, the reaction was catalyzed by free lipases in a batch reactor and the influ...

  16. Biodiesel production with special emphasis on lipase-catalyzed transesterification.

    Science.gov (United States)

    Bisen, Prakash S; Sanodiya, Bhagwan S; Thakur, Gulab S; Baghel, Rakesh K; Prasad, G B K S

    2010-08-01

    The production of biodiesel by transesterification employing acid or base catalyst has been industrially accepted for its high conversion and reaction rates. Downstream processing costs and environmental problems associated with biodiesel production and byproducts recovery have led to the search for alternative production methods. Recently, enzymatic transesterification involving lipases has attracted attention for biodiesel production as it produces high purity product and enables easy separation from the byproduct, glycerol. The use of immobilized lipases and immobilized whole cells may lower the overall cost, while presenting less downstream processing problems, to biodiesel production. The present review gives an overview on biodiesel production technology and analyzes the factors/methods of enzymatic approach reported in the literature and also suggests suitable method on the basis of evidence for industrial production of biodiesel.

  17. Synthesis of Rosin Acid Starch Catalyzed by Lipase

    Directory of Open Access Journals (Sweden)

    Rihui Lin

    2014-01-01

    Full Text Available Rosin, an abundant raw material from pine trees, was used as a starting material directly for the synthesis of rosin acid starch. The esterification reaction was catalyzed by lipase (Novozym 435 under mild conditions. Based on single factor experimentation, the optimal esterification conditions were obtained as follows: rosin acid/anhydrous glucose unit in the molar ratio 2 : 1, reaction time 4 h at 45°C, and 15% of lipase dosage. The degree of substitution (DS reaches 0.098. Product from esterification of cassava starch with rosin acid was confirmed by FTIR spectroscopy and iodine coloration analysis. Scanning electron microscopy and X-ray diffraction analysis showed that the morphology and crystallinity of the cassava starch were largely destroyed. Thermogravimetric analysis indicated that thermal stability of rosin acid starch decreased compared with native starch.

  18. Gelatin blends with alginate: gels for lipase immobilization and purification.

    Science.gov (United States)

    Fadnavis, Nitin W; Sheelu, Gurrala; Kumar, Bezavada Mani; Bhalerao, Mahendra U; Deshpande, Ashlesha A

    2003-01-01

    Blends of natural polysaccharide sodium alginate (5%) with gelatin (3%) cross-linked with glutaraldehyde provide beads with excellent compressive strength (8 x 10(4) Pa) and regular structure on treatment with calcium chloride. Lipases from porcine pancreas, Pseudomonas cepacia, and Candida rugosa were immobilized in such a blend with excellent efficiency. The immobilized enzymes were stable and were reused several times without significant loss of enzyme activity both in aqueous and reverse micellar media. The beads were functionalized with succinic anhydride to obtain beads with extra carboxylic acid groups. These functionalized beads were then successfully used for 7.4-fold purification of crude porcine pancreatic lipase in a simple operation of protein binding at pH 5 and release at pH 8.5.

  19. Covalent immobilization of Pseudomonas cepacia lipase on semiconducting materials

    Energy Technology Data Exchange (ETDEWEB)

    Fernandez, Renny Edwin [Microelectronics and MEMS Laboratory, Department of Electrical Engineering, Indian Institute of Technology Madras, Chennai (India)], E-mail: rennyedwin@gmail.com; Bhattacharya, Enakshi [Microelectronics and MEMS Laboratory, Department of Electrical Engineering, Indian Institute of Technology Madras, Chennai (India)], E-mail: enakshi@ee.iitm.ac.in; Chadha, Anju [Department of Biotechnology, National Centre for Catalysis Research, Indian Institute of Technology Madras, Chennai (India)], E-mail: anjuc@iitm.ac.in

    2008-05-30

    Lipase from Pseudomonas cepacia was covalently immobilized on crystalline silicon, porous silicon and silicon nitride surfaces. The various stages of immobilization were characterized using FTIR (Fourier transform infrared) spectroscopy. The surface topography of the enzyme immobilized surfaces was investigated using scanning electron microscopy (SEM). The quantity of the immobilized active enzyme was estimated by the para-nitrophenyl palmitate (pNPP) assay. The immobilized lipase was used for triglyceride hydrolysis and the acid produced was detected by a pH sensitive silicon nitride surface as a shift in the C-V (capacitance-voltage) characteristics of an electrolyte-insulator-semiconductor capacitor (EISCAP) thus validating the immobilization method for use as a biosensor.

  20. LIPASE IMMOBILIZED MEMBRANE REACTOR APPLIED TO BABASSU OIL HYDROLYSIS

    Directory of Open Access Journals (Sweden)

    Merçon F.

    1997-01-01

    Full Text Available This work deals with enzymatic hydrolysis of babassu oil by immobilized lipase in membrane reactors of two types: a flat plate nylon membrane and a hollow fiber polyetherimide membrane on which surface commercial lipases were immobilized by adsorption. Experiments conducted in the hollow fiber reactor showed that during the immobilization step enzyme adsorption followed a sigmoid model, with a maximum adsorption equilibrium time of 30 minutes. Concerning the hydrodynamics of the liquid phases, the results indicate that main diffusional limitations occurred in the organic phase. The amount of protein immobilized and the maximum productivity were, respectively, 1.97 g/m2 and 44 m molH+/m2.s for the hollow fiber and 1.2 g/m2 and 56 m molH+/m2.s for the flat and plate membrane. Both reactors were able to perform the hydrolysis reaction, while maintaining absolute separation of the two phases by the membrane

  1. Enzymatic Production of FAME Biodiesel with Soluble Lipases

    DEFF Research Database (Denmark)

    T. Gundersen, Maria; Heltborg, Carsten Kirstejn; Yang, V

    Biodiesel is a viable alternative to fossil fuels, and biocatalysis is gaining interest as a greener process. We focus on converting oils to Fatty Acid Methyl Ester (FAME) using soluble lipases, which offer an advantage compared to immobilized enzymes by cost efficiency and ease of implementation.......p.) of certain oils, which is not compatible with the temperature range where lipases are most active. To address this, here we explored a novel production strategy that accommodates the enzymatic requirements with the chemical limits of the substrates. The m.p. of the methyl ester product is lower than...... that of the starting material. Thus, we have incorporated a varying amount of the product to lower the m.p. of the starting material. Our case study is the reaction of Palm Fatty Acid Distillate (PFAD) to FAME. Conversion rates have been measured with varying temperatures, water concentration, and initial methanol...

  2. Regioselective Alcoholysis of Silychristin Acetates Catalyzed by Lipases

    Directory of Open Access Journals (Sweden)

    Eva Vavříková

    2015-05-01

    Full Text Available A panel of lipases was screened for the selective acetylation and alcoholysis of silychristin and silychristin peracetate, respectively. Acetylation at primary alcoholic group (C-22 of silychristin was accomplished by lipase PS (Pseudomonas cepacia immobilized on diatomite using vinyl acetate as an acetyl donor, whereas selective deacetylation of 22-O-acetyl silychristin was accomplished by Novozym 435 in methyl tert-butyl ether/ n-butanol. Both of these reactions occurred without diastereomeric discrimination of silychristin A and B. Both of these enzymes were found to be capable to regioselective deacetylation of hexaacetyl silychristin to afford penta-, tetra- and tri-acetyl derivatives, which could be obtained as pure synthons for further selective modifications of the parent molecule.

  3. Enzymatic production of alkyl esters through alcoholysis: A critical evaluation of lipases and alcohols

    DEFF Research Database (Denmark)

    Li, Deng; Xu, Xuebing; Gudmundur G, Haraldsson

    2005-01-01

    yield of FA alkyl esters, with yields well over 90% for methanol, absolute ethanol, and 1-propanol. Overall, 96% ethanol was the preferred alcohol for all lipases except Novozym 435, and ethanolysis reactions reached the maximal conversion efficiency. Increasing the water content in the system resulted...... in an increased degree of conversion for all lipases except Novozym 435. The secondary alcohol 2-propanol significantly reduced the alcoholysis reaction with all lipases; however, the branch-chain isobutanol was more advantageous than linear 1-butanol for Novozym 435, Lipozyme RM IM, and Lipase PS-C. Many...

  4. Lipase entrapment in PVA/Chitosan biodegradable film for reactor coatings

    Energy Technology Data Exchange (ETDEWEB)

    Batista, Karla A. [Departamento de Bioquímica e Biologia Molecular, Laboratório de Química de Proteínas, Universidade Federal de Goiás, Cx. Postal 131, 74001-970, Goiânia, GO (Brazil); Lopes, Flavio Marques [Departamento de Bioquímica e Biologia Molecular, Laboratório de Química de Proteínas, Universidade Federal de Goiás, Cx. Postal 131, 74001-970, Goiânia, GO (Brazil); Unidade Universitária de Ciências Exatas e Tecnológicas, Universidade Estadual de Goiás, Anápolis, GO (Brazil); Yamashita, Fabio [Departamento de Tecnologia de Alimentos e Medicamentos, Laboratório de Tecnologia, Universidade Estadual de Londrina, Cx. Postal 6001, CEP 86051-990, Londrina, PR (Brazil); Fernandes, Kátia Flávia, E-mail: katia@icb.ufg.br [Departamento de Bioquímica e Biologia Molecular, Laboratório de Química de Proteínas, Universidade Federal de Goiás, Cx. Postal 131, 74001-970, Goiânia, GO (Brazil)

    2013-04-01

    This study reports the development and characterization of novel biodegradable film, based on chitosan and polyvinyl alcohol containing lipase entrapped. The films showed a thickness of 70.4 and 79 μm to PVA/Chitosan and PVA/Chitosan/Lipase, respectively. The entrapment of lipase in PVA/Chitosan film resulted in increasing of 69.4% tensile strength (TS), and 52.4% of elongation. SEM images showed the formation of a continuous film, without pores or cracks. The lipase entrapment efficiency was estimated in 92% and the films were repeatedly used for 25 hydrolytic cycles, maintaining 62% of initial activity. The PVA/Chitosan/Lipase film was used for olive oil hydrolysis of high performance. These results indicate that PVA/Chitosan/Lipase is a promising material for biotechnology applications such as triacylglycerol hydrolysis and biodiesel production. - Highlights: ► Development and characterization of PVA/Chitosan biodegradable film ► Lipase immobilization onto PVA/Chitosan film ► PVA/Chitosan/Lipase film for reactor coating ► Olive oil hydrolysis using PVA/Chitosan/Lipase film.

  5. Extraction and Characteristics of Anti-obesity Lipase Inhibitor from Phellinus linteus.

    Science.gov (United States)

    Lee, Jong-Kug; Jang, Jeong-Hoon; Lee, Jong-Tae; Lee, Jong-Soo

    2010-03-01

    To develop a potent anti-obesity lipase inhibitor from mushroom, the lipase inhibitory activities of various mushroom extracts were determined. Methanol extracts from Phellinus linteus fruiting body exhibited the highest lipase inhibitory activity (72.8%). The inhibitor was maximally extracted by treatment of a P. linteus fruiting body with 80% methanol at 40℃ for 24 hr. After partial purification by systematic solvent extraction, the inhibitor was stable in the range of 40~80℃ and pH 2.0~9.0. In addition to lipase inhibitory activity, the inhibitor showed 59.4% of superoxide dismutase-like activity and 56.3% of acetylcholinesterase inhibitory activity.

  6. Stability studies of immobilized lipase on rice husk and eggshell membrane

    Science.gov (United States)

    Abdulla, R.; Sanny, S. A.; Derman, E.

    2017-06-01

    Lipase immobilization for biodiesel production is gaining importance day by day. In this study, lipase from Burkholderia cepacia was immobilized on activated support materials namely rice husk and egg shell membrane. Both rice husk and eggshell membrane are natural wastes that holds a lot of potential as immobilization matrix. Rice husk and eggshell membrane were activated with glutaraldehyde. Lipase was immobilized on the glutaraldehyde-activated support material through adsorption. Immobilization efficiency together with enzyme activity was observed to choose the highest enzyme loading for further stability studies. Immobilization efficiency of lipase on rice husk was 81 as compared to an immobilization efficiency of 87 on eggshell membrane. Immobilized lipase on eggshell membrane exhibited higher enzyme activity as compared to immobilized lipase on rice husk. Eggshell membrane also reported higher stability than rice husk as immobilization matrix. Both types of immobilized lipase retatined its activity after ten cycles of reuse. In short, eggshell membrane showed to be a better immobilization platform for lipase as compared to rice husk. However, with further improvement in technique of immobilization, the stability of both types of immobilized lipase can be improved to a greater extent.

  7. Acid Lipase from Candida viswanathii: Production, Biochemical Properties, and Potential Application

    Science.gov (United States)

    de Almeida, Alex Fernando; Carmona, Eleonora Cano

    2013-01-01

    Influences of environmental variables and emulsifiers on lipase production of a Candida viswanathii strain were investigated. The highest lipase activity (101.1 U) was observed at 210 rpm, pH 6.0, and 27.5°C. Other fermentation parameters analyzed showed considerable rates of biomass yield (Y L/S = 1.381 g/g), lipase yield (Y L/S = 6.892 U/g), and biomass productivity (P X = 0.282 g/h). Addition of soybean lecithin increased lipase production in 1.45-fold, presenting lipase yield (Y L/S) of 10.061 U/g. Crude lipase presented optimal activity at acid pH of 3.5, suggesting a new lipolytic enzyme for this genus and yeast in general. In addition, crude lipase presented high stability in acid conditions and temperature between 40 and 45°C, after 24 h of incubation in these temperatures. Lipase remained active in the presence of organic solvents maintaining above 80% activity in DMSO, methanol, acetonitrile, ethanol, acetone, 1-propanol, isopropanol, and 2-propanol. Effectiveness for the hydrolysis of a wide range of natural triglycerides suggests that this new acid lipase has high potential application in the oleochemical and food industries for hydrolysis and/or modification of triacylglycerols to improve the nutritional properties. PMID:24350270

  8. Lipase cocktail for efficient conversion of oils containing phospholipids to biodiesel.

    Science.gov (United States)

    Amoah, Jerome; Ho, Shih-Hsin; Hama, Shinji; Yoshida, Ayumi; Nakanishi, Akihito; Hasunuma, Tomohisa; Ogino, Chiaki; Kondo, Akihiko

    2016-07-01

    The presence of phospholipid has been a challenge in liquid enzymatic biodiesel production. Among six lipases that were screened, lipase AY had the highest hydrolysis activity and a competitive transesterification activity. However, it yielded only 21.1% FAME from oil containing phospholipids. By replacing portions of these lipases with a more robust bioFAME lipase, CalT, the combination of lipase AY-CalT gave the highest FAME yield with the least amounts of free fatty acids and partial glycerides. A higher methanol addition rate reduced FAME yields for lipase DF-CalT and A10D-CalT combinations while that of lipase AY-CalT combination improved. Optimizing the methanol addition rate for lipase AY-CalT resulted in a FAME yield of 88.1% at 2h and more than 95% at 6h. This effective use of lipases could be applied for the rapid and economic conversion of unrefined oils to biodiesel.

  9. Structure prediction and functional analyses of a thermostable lipase obtained from Shewanella putrefaciens.

    Science.gov (United States)

    Khan, Faez Iqbal; Nizami, Bilal; Anwer, Razique; Gu, Ke-Ren; Bisetty, Krishna; Hassan, Md Imtaiyaz; Wei, Dong-Qing

    2017-08-01

    Previous experimental studies on thermostable lipase from Shewanella putrefaciens suggested the maximum activity at higher temperatures, but with little information on its conformational profile. In this study, the three-dimensional structure of lipase was predicted and a 60 ns molecular dynamics (MD) simulation was carried out at temperatures ranging from 300 to 400 K to better understand its thermostable nature at the molecular level. MD simulations were performed in order to predict the optimal activity of thermostable lipase. The results suggested strong conformational temperature dependence. The thermostable lipase maintained its bio-active conformation at 350 K during the 60 ns MD simulations.

  10. Influence of dietary recombinant microbial lipase on performance and quality characteristics of rainbow trout, Oncorhynchus mykiss

    DEFF Research Database (Denmark)

    Samuelsen, Troels; Isaksen, Mai; McLean, Ewen

    2001-01-01

    In order to assess whether supplementary lipase affected growth and body composition of trout, four diets were produced, consisting of (A) feed containing high (2083 mg kg(-1)), (B) low (208.3 mg kg(-1)) concentrations of lipase, (C) heat-treated (inactivated) lipase (2083 mg kg(-1)), and (D...... higher(P 0.05) on growth, fillet proximate composition, hepatosomatic, cardiac, or gut indices, and carcass percentage. However, lipase supplementation influenced the mono-unsaturated fatty acid profiles of the fillet (P

  11. Lipases production by solid-state fermentation: the case of Rhizopus homothallicus in perlite.

    Science.gov (United States)

    Velasco-Lozano, Susana; Volke-Sepulveda, Tania; Favela-Torres, Ernesto

    2012-01-01

    Lipases are widely used in the industry for different purposes. Although these enzymes are mainly produced by submerged fermentation, lipase production by solid-state fermentation (SSF) has been gaining interest due to the advantages of this type of culture. Major advantages are higher production titers and productivity, less catabolite repression, and use of the dried fermented material as biocatalyst. This chapter describes a traditional methodology to produce fungal (Rhizopus homothallicus) lipases by SSF using perlite as inert support. The use of different devices (glass columns or Erlenmeyer flasks) and type of inoculum (spores or growing mycelium) is considered so that lipase production by SSF could be easily performed in any laboratory.

  12. Lipase NS81006 immobilized on Fe3O4 magnetic nanoparticles for biodiesel production

    Directory of Open Access Journals (Sweden)

    Thangaraj Baskar

    2016-06-01

    Full Text Available Lipase-catalyzed biodiesel production is being the object of extensive research due to the demerits of chemical based catalytic system. Lipase immobilized on Fe3O4 magnetic nanoparticles has the integrated advantages of traditional immobilized lipase and free lipase for its rather fast reaction rate and easy separation. It has been demonstrated that free lipase NS81006 has potential in catalyzing the alcoholysis of renewable oils for biodiesel preparation. In this study, Fe3O4 magnetic nanoparticles functionalized with organosilane compounds like (3-aminopropyltriethyloxysilane (APTES and (3-mercaptopropyltrimethoxysilane MPTMS were used as carriers for lipase immobilization. Lipase NS81006 was covalently bound to the organosilane-functionalized magnetic nanoparticles by using glutaraldehyde cross-linking reagent. A biodiesel yield of 89% and 81% could be achieved by lipase immobilized on APTES-Fe3O4 and MPTMS-Fe3O4 magnetic nanoparticles respectively under optimized conditions of oil to methanol molar ratio 1:3 with three step addition of methanol, reaction temperature 45°C and reaction time duration 12 h. The lipases immobilized on magnetic nanoparticles could be recovered easily by external magnetic field for further use.

  13. In situ visualization and effect of glycerol in lipase-catalyzed ethanolysis of rapeseed oil

    DEFF Research Database (Denmark)

    Xu, Yuan; Nordblad, Mathias; Nielsen, Per M.;

    2011-01-01

    Immobilized lipases can be used in biodiesel production to overcome many disadvantages of the conventional base-catalyzed process. However, the glycerol by-product poses a potential problem for the biocatalytic process as it is known to inhibit immobilized lipases, most likely by clogging...... to illustrate the interaction of glycerol with immobilized lipases and thus provided an aid for screening supports for lipase immobilization according to their interaction with glycerol. Glycerol was found to have great affinity for silica, less for polystyrene and no affinity for supports made from...

  14. Molecular and functional diversity of yeast and fungal lipases: their role in biotechnology and cellular physiology.

    Science.gov (United States)

    Gupta, Rani; Kumari, Arti; Syal, Poonam; Singh, Yogesh

    2015-01-01

    Lipase catalyzes hydrolysis of fats in lipid water interphase and perform variety of biotransformation reactions under micro aqueous conditions. The major sources include microbial lipases; among these yeast and fungal lipases are of special interest because they can carry out various stereoselective reactions. These lipases are highly diverse and are categorized into three classes on the basis of oxyanion hole: GX, GGGX and Y. The detailed phylogenetic analysis showed that GX family is more diverse than GGGX and Y family. Sequence and structural comparisons revealed that lipases are conserved only in the signature sequence region. Their characteristic structural determinants viz. lid, binding pocket and oxyanion hole are hotspots for mutagenesis. Few examples are cited in this review to highlight the multidisciplinary approaches for designing novel enzyme variants with improved thermo stability and substrate specificity. In addition, we present a brief account on biotechnological applications of lipases. Lipases have also gained attention as virulence factors, therefore, we surveyed the role of lipases in yeast physiology related to colonization, adhesion, biofilm formation and pathogenesis. The new genomic era has opened numerous possibilities to genetically manipulate lipases for food, fuel and pharmaceuticals.

  15. Adverse effect of chloride impurities on lipase-catalyzed transesterifications in ionic liquids.

    Science.gov (United States)

    Lee, Sang Hyun; Ha, Sung Ho; Lee, Sun Bok; Koo, Yoon-Mo

    2006-09-01

    The adverse influence of chloride impurities on the lipase-catalyzed transesterification in ionic liquid is described. The activity of lipase from Rhizomucor miehei exponentially decreased with increasing Cl(-) content in 1-octyl-3-methylimidazolium bis[(trifluoromethyl)sulfonyl] amide, [Omim][Tf(2)N], and the activity of lipase in [Omim][Tf(2)N] mixture containing 2% [Omim] [Cl] was only about 2% of the activity in pure [Omim][Tf(2)N]. The activity of lipase from Candidantarctica linearly decreased at about 5% with every 1% increase in [Omim][Cl] with there being no activity in [Omim][Tf(2)N] containing about 20% [Omim][Cl].

  16. Factors influencing the activity and thermostability of immobilized porcine pancreatic lipase.

    Science.gov (United States)

    Kéry, V; Haplová, J; Tihlárik, K; Schmidt, S

    1990-01-01

    Lipase from porcine pancreas was immobilized on cellulose beads having various degrees of hydrophobicity, by covalent linking and by hydrophobic adsorption. Lipolytic activity was measured in heterogeneous organic-aqueous systems of various hydrophobicities using olive oil as a substrate. The main factors influencing lipase activity were hydrophobicity of the reaction mixture and of the carrier. Carriers with increased hydrophobicity enhanced lipase activity more than less hydrophobic ones. Lipase immobilized covalently on cellulose beads was less active than that adsorbed onto tritylcellulose but was considerably more thermostable.

  17. Lipase specificity towards eicosapentaenoic acid and docosahexaenoic acid depends on substrate structure.

    Science.gov (United States)

    Lyberg, Ann-Marie; Adlercreutz, Patrick

    2008-02-01

    The fatty acid specificity of five lipases towards eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) was evaluated in the hydrolysis of fish oil, squid oil and a model system. The model system contained methyl esters of EPA, DHA and palmitic acid. All the investigated lipases discriminated against both EPA and DHA more in the model system than in the natural oils. Thus both EPA and DHA were more easily hydrolysed from a glyceride than from a methyl ester. In the model system, the lipase from Candida rugosa showed the highest discrimination against DHA, while the lipases from Pseudomonas fluorescens and Pseudomonas cepacia discriminated against EPA the most. In a glyceride, the fatty acid specificity of lipases towards EPA and DHA was affected by the positional distribution of the fatty acids and the glyceride structure due to the regiospecificity and triglyceride specificity of the lipase. In the oils, the Pseudomonas lipases also discriminated against EPA the most, while DHA was initially discriminated the most by the lipase from Thermomyces lanuginosus. However, after longer reaction times the enrichment of DHA in the glyceride fraction of the oils was greatest for the lipase from C. rugosa.

  18. Acid lipase from Candida viswanathii: production, biochemical properties, and potential application.

    Science.gov (United States)

    de Almeida, Alex Fernando; Tauk-Tornisielo, Sâmia Maria; Carmona, Eleonora Cano

    2013-01-01

    Influences of environmental variables and emulsifiers on lipase production of a Candida viswanathii strain were investigated. The highest lipase activity (101.1 U) was observed at 210 rpm, pH 6.0, and 27.5°C. Other fermentation parameters analyzed showed considerable rates of biomass yield (Y L/S = 1.381 g/g), lipase yield (Y L/S = 6.892 U/g), and biomass productivity (P X = 0.282 g/h). Addition of soybean lecithin increased lipase production in 1.45-fold, presenting lipase yield (Y L/S ) of 10.061 U/g. Crude lipase presented optimal activity at acid pH of 3.5, suggesting a new lipolytic enzyme for this genus and yeast in general. In addition, crude lipase presented high stability in acid conditions and temperature between 40 and 45°C, after 24 h of incubation in these temperatures. Lipase remained active in the presence of organic solvents maintaining above 80% activity in DMSO, methanol, acetonitrile, ethanol, acetone, 1-propanol, isopropanol, and 2-propanol. Effectiveness for the hydrolysis of a wide range of natural triglycerides suggests that this new acid lipase has high potential application in the oleochemical and food industries for hydrolysis and/or modification of triacylglycerols to improve the nutritional properties.

  19. Role of lipase in Burkholderia cepacia complex (Bcc) invasion of lung epithelial cells.

    Science.gov (United States)

    Mullen, T; Markey, K; Murphy, P; McClean, S; Callaghan, M

    2007-12-01

    The Burkholderia cepacia complex (Bcc) is a group of ten closely related species associated with life-threatening infection in cystic fibrosis (CF). These bacteria are highly antibiotic resistant, with some strains transmissible, and in a subgroup of patients, they can cause a rapid and fatal necrotising pneumonia. The Bcc organisms produce a range of exoproducts with virulence potential, including exopolysaccharide, proteases and lipases. Many members of the Bcc are also capable of epithelial cell invasion, although the mechanism(s) involved are poorly understood. This study investigates a role for Bcc lipase in epithelial cell invasion by Bcc strains. Lipase activity was measured in eight species of the Bcc. Strains that produced high levels of lipase were predominantly from the B. multivorans and B. cenocepacia species. Pre-treatment of two epithelial cell lines with Bcc lipase significantly increased invasion by two B. multivorans strains and one B. cenocepacia strain and did not affect either plasma membrane or tight junction integrity. Inhibition of Bcc lipase production by the lipase inhibitor Orlistat significantly decreased invasion by both B. multivorans and B. cenocepacia strains in a concentration-dependent manner. This study demonstrates the extent of lipase production across the Bcc and establishes a potential role for lipase in Bcc epithelial cell invasion.

  20. Cold-adapted organic solvent tolerant alkalophilic family I.3 lipase from an Antarctic Pseudomonas.

    Science.gov (United States)

    Ganasen, Menega; Yaacob, Norhayati; Rahman, Raja Noor Zaliha Raja Abd; Leow, Adam Thean Chor; Basri, Mahiran; Salleh, Abu Bakar; Ali, Mohd Shukuri Mohamad

    2016-11-01

    Lipolytic enzymes with cold adaptation are gaining increasing interest due to their biotechnological prospective. Previously, a cold adapted family I.3 lipase (AMS8 lipase) was isolated from an Antarctic Pseudomonas. AMS8 lipase was largely expressed in insoluble form. The refolded His-tagged recombinant AMS8 lipase was purified with 23.0% total recovery and purification factor of 9.7. The purified AMS8 lipase migrated as a single band with a molecular weight approximately 65kDa via electrophoresis. AMS8 lipase was highly active at 30°C at pH 10. The half-life of AMS8 lipase was reported at 4 and 2h under the incubation of 30 and 40°C, respectively. The lipase was stable over a broad range of pH. It showed enhancement effect in its relative activity under the presence of Li(+), Na(+), K(+), Rb(+) and Cs(+) after 30min treatment. Heavy metal ions such as Cu(2+), Fe(3+) and Zn(2+) inhibited AMS8 activity. This cold adapted alkalophilic AMS lipase was also active in various organic solvent of different polarity. These unique properties of this biological macromolecule will provide considerable potential for many biotechnological applications and organic synthesis at low temperature.