Sample records for c-catalyzed sphingomyelin hydrolysis

  1. Phospholipase C-catalyzed sphingomyelin hydrolysis in a membrane reactor for ceramide production

    Zhang, Long; Liang, Shanshan; Hellgren, Lars; Jonsson, Gunnar Eigil; Xu, Xuebing


    A membrane reactor for the production of ceramide through sphingomyelin hydrolysis with phospholipase C from Clostridium perfringens was studied for the first time. Ceramide has raised a large interest as an active component in both pharmaceutical and cosmetic industry. The enzymatic hydrolysis o...... study demonstrated the improved enzyme reusability, the fast immobilization process, the straightforward up-scaling and the combination of the hydrolysis with the product separation in the membrane reactor developed....

  2. Kinetic study of sphingomyelin hydrolysis for ceramide production

    Zhang, Long; Hellgren, Lars; Xu, Xuebing


    reaction at 65 °C. The reactions for both the soluble and immobilized enzymes followed Michaelis–Menten kinetics. Km's for the soluble and immobilized enzymes were 1.07 ± 0.32 and 1.26 ± 0.19 mM, respectively. The value of Vmax was markedly decreased by the immobilization without much change in Km, as if...... cosmetic and pharmaceutical industries such as in hair and skin care products. The enzymatic hydrolysis of sphingomyelin has been proved to be a feasible method to produce ceramide. The kinetic performance of sphingomyelin hydrolysis in the optimal two-phase (water:organic solvent) reaction system was...... investigated to elucidate the possible reaction mechanism and also to further improve the hydrolysis performance. Enzyme in solution had less thermal stability than the enzyme powder and the immobilized enzyme. The thermal inactivation of phospholipase C in all the three forms did not follow the first order...

  3. Immobilization of phospholipase C for the production of ceramide from sphingomyelin hydrolysis

    Zhang, Long; Hellgren, Lars; Xu, Xuebing


    The immobilization of Clostridium perfringens phospholipase C was studied for the first time and the catalytic properties of the immobilized enzyme were investigated for the hydrolysis of sphingomyelin to produce ceramide. Ceramide is of great commercial potentials in cosmetic and pharmaceutical...... industries such as in hair and skin care products, due to its major role in maintaining the water-retaining properties of the epidermis. The feasibility of enzymatic production of ceramide through hydrolysis of sphingomyelin has previously been proven. In order to improve the reusability of the enzyme, the...

  4. Immobilization of phospholipase C for the production of ceramide from sphingomyelin hydrolysis

    Zhang, Long; Hellgren, Lars; Xu, Xuebing


    The immobilization of Clostridium perfringens phospholipase C was studied for the first time and the catalytic properties of the immobilized enzyme were investigated for the hydrolysis of sphingomyelin to produce ceramide. Ceramide is of great commercial potentials in cosmetic and pharmaceutical...

  5. Enzymatic Modification of Sphingomyelin

    Zhang, Long; Hellgren, Lars; Xu, Xuebing

    -efficient, high yield production methods are of great interest. In the present study, the potential of producing ceramide through enzymatic hydrolysis of sphingomyelin have been studied. Sphingomyelin (SM) is a ubiquitous membrane-lipid and dairy products or by-products is a rich source of sphingomyelin. In...

  6. Quantitative Analysis of Sphingomyelin by High-Performance Liquid Chromatography after Enzymatic Hydrolysis

    Seunghyun Lee


    Full Text Available Sphingomyelin is the most abundant sphingolipid in mammalian cells and is mostly present in the plasma membrane. A new analytical method using high-performance liquid chromatography (HPLC was developed to quantify sphingomyelin in mouse plasma and tissues, 3T3-L1 cells, rat aortic smooth muscle cells, and HT-29 cells. Sphingomyelin and dihydrosphingomyelin, an internal standard, were separated by high-performance thin-layer chromatography and simultaneously hydrolyzed with sphingolipid ceramide N-deacylase and sphingomyelinase to release sphingosine and dihydrosphingosine, respectively. Sphingomyelin content was measured by HPLC following o-phthalaldehyde derivatization. Sphingomyelin concentrations in 3T3-L1 cells, rat aortic smooth muscle cells, and HT-29 cells were 60.10±0.24, 62.69±0.08, and 58.38±0.37 pmol/μg protein, respectively, whereas those in brain, kidney, and liver of ICR mice were 55.60±0.43, 43.75±0.21, and 22.26±0.14 pmol/μg protein. The sphingomyelin concentration in mouse plasma was 407.40±0.31 μM. The limits of detection and quantification for sphingomyelin were 5 and 20 pmol, respectively, in the HPLC analysis with fluorescence detection. This sensitivity was sufficient for analyzing sphingomyelin in biological samples. In conclusion, this analytical method is a sensitive and specific technique for quantifying sphingomyelin and was successfully applied to diverse biological samples with excellent reproducibility.

  7. Enzymatic Modification of Sphingomyelin

    -efficient, high yield production methods are of great interest. In the present study, the potential of producing ceramide through the enzymatic hydrolysis of sphingomyelin have been studied. sphingomyelin is a ubiquitous membrane-lipid and rich in dairy products or by-products. It has been verified that...... sphingomyelin modification gives a feasible approach to the potential production of ceramide. The reaction system has been improved through system evaluation and the optimization of several important factors, and phospholipase C from Clostridium perfringens shows higher activity towards the hydrolysis reaction...

  8. Enzymatic Modification of Sphingomyelin

    Zhang, Long; Hellgren, Lars; Xu, Xuebing

    Due to its major role in maintaining the water-retaining properties of the epidermis, ceramide is of great commercial potential in cosmetic and pharmaceuticals such as hair and skin care products. Currently, chemical synthesis of ceramide is a costly process, and developments of alternative cost......-efficient, high yield production methods are of great interest. In the present study, the potential of producing ceramide through enzymatic hydrolysis of sphingomyelin have been studied. Sphingomyelin (SM) is a ubiquitous membrane-lipid and dairy products or by-products is a rich source of sphingomyelin. In...

  9. Enzymatic Modification of Sphingomyelin

    Zhang, Long; Hellgren, Lars; Xu, Xuebing

    Sphingomyelin (SM) is a ubiquitous component of animal cell membranes, and it is the most abundant sphingolipid. Ceramide, a hydrolysis product from SM, has an important role in cellular signaling, and especially in the regulation of apoptosis, cell differentiation, transformation and proliferation....... Therefore, it is desirable to develop alternative cost-efficient, high yield production methods. This study optimized the enzymatic production of ceramide from SM. Phospholipase C from Clostridium perfringens was chosen to catalyze the reaction. Several important factors were considered in optimization....

  10. Regulation of sphingomyelin metabolism.

    Bienias, Kamil; Fiedorowicz, Anna; Sadowska, Anna; Prokopiuk, Sławomir; Car, Halina


    Sphingolipids (SFs) represent a large class of lipids playing diverse functions in a vast number of physiological and pathological processes. Sphingomyelin (SM) is the most abundant SF in the cell, with ubiquitous distribution within mammalian tissues, and particularly high levels in the Central Nervous System (CNS). SM is an essential element of plasma membrane (PM) and its levels are crucial for the cell function. SM content in a cell is strictly regulated by the enzymes of SM metabolic pathways, which activities create a balance between SM synthesis and degradation. The de novo synthesis via SM synthases (SMSs) in the last step of the multi-stage process is the most important pathway of SM formation in a cell. The SM hydrolysis by sphingomyelinases (SMases) increases the concentration of ceramide (Cer), a bioactive molecule, which is involved in cellular proliferation, growth and apoptosis. By controlling the levels of SM and Cer, SMSs and SMases maintain cellular homeostasis. Enzymes of SM cycle exhibit unique properties and diverse tissue distribution. Disturbances in their activities were observed in many CNS pathologies. This review characterizes the physiological roles of SM and enzymes controlling SM levels as well as their involvement in selected pathologies of the Central Nervous System, such as ischemia/hypoxia, Alzheimer disease (AD), Parkinson disease (PD), depression, schizophrenia and Niemann Pick disease (NPD). PMID:26940196

  11. Enzymatic Production of Ceramide from Sphingomyelin

    Zhang, Long; Hellgren, Lars; Xu, Xuebing

    Ceramide is the key intermediate in the biosynthesis of all complex sphingolipids. Due to its major role in maintaining the water-retaining properties of the epidermis, ceramide is of great commercial potential in cosmetic and pharmaceuticals such as hair and skin care products. Currently, chemical...... contains a ceramide moiety, is a ubiquitous component of animal cell membranes, and dairy products or by-products is a rich source of sphingomyelin. It has been verified that enzymatic modification of sphingomyelin is a feasible approach for production of ceramide. The reaction system has been optimized...... through system evaluation and the optimization of several important factors. Sphingomyelin hydrolysis proved to be more efficient in two-phase (water: organic solvent) system than in one-phase (water-saturated organic solvent) system. Phospholipase C from Clostridium perfringens is the tested enzyme which...

  12. Inhibitors of the sphingomyelin cycle: Sphingomyelin synthases and sphingomyelinases.

    Adada, Mohamad; Luberto, Chiara; Canals, Daniel


    Sphingolipids are a class of bioactive lipids, which are key modulators of an increasing number of physiologic and pathophysiologic processes that include cell cycle, apoptosis, angiogenesis, stress and inflammatory responses. Sphingomyelin is an important structural component of biological membranes, and one of the end-points in the synthesis of sphingolipids. Mainly synthetized in the Golgi apparatus, sphingomyelin is transported to all other biological membranes. Upon stimulation, sphingomyelin can be hydrolyzed to ceramide by 5 different sphingomyelinases. The diversity and cellular topology of ceramide allow it to exert multiple biologies. Furthermore, ceramide can be metabolized to many other bioactive sphingolipids. Ceramide, coming from sphingomyelin or other complex sphingolipids, can be hydrolyzed to sphingosine, which can easily change cellular localization. In turn, sphingosine can be recycled to ceramide and to sphingomyelin in the endoplasmic reticulum, completing the sphingomyelin cycle. Our understanding of the roles of various sphingolipids in the regulation of different cellular processes has come from studying the enzymes that regulate these sphingolipids, and their manipulation. The use of pharmacologic inhibitors has been critical for their study, as well as being promising bullets for disease treatment. Some of these diseases involving the sphingomyelin cycle include cancer, inflammation, atherosclerosis, diabetes and some rare diseases such as Niemann-Pick disease. This review will focus on the enzymes involved in the sphingomyelin cycle, their history, and their involvement in pathophysiological processes. Finally, it will describe in details all the small molecules that are being used to inhibit these enzymes and their use in therapeutics. PMID:26200918

  13. Cytochrome c catalyzes the in vitro synthesis of arachidonoyl glycine

    Long chain fatty acyl glycines are an emerging class of biologically active molecules that occur naturally and produce a wide array of physiological effects. Their biosynthetic pathway, however, remains unknown. Here we report that cytochrome c catalyzes the synthesis of N-arachidonoyl glycine (NAGly) from arachidonoyl coenzyme A and glycine in the presence of hydrogen peroxide. The identity of the NAGly product was verified by isotope labeling and mass analysis. Other heme-containing proteins, hemoglobin and myoglobin, were considerably less effective in generating arachidonoyl glycine as compared to cytochrome c. The reaction catalyzed by cytochrome c in vitro points to its potential role in the formation of NAGly and other long chain fatty acyl glycines in vivo

  14. Sphingomyelin Synthases Regulate Protein Trafficking and Secretion

    Subathra, Marimuthu; Qureshi, Asfia; Luberto, Chiara


    Sphingomyelin synthases (SMS1 and 2) represent a class of enzymes that transfer a phosphocholine moiety from phosphatidylcholine onto ceramide thus producing sphingomyelin and diacylglycerol (DAG). SMS1 localizes at the Golgi while SMS2 localizes both at the Golgi and the plasma membrane. Previous studies from our laboratory showed that modulation of SMS1 and, to a lesser extent, of SMS2 affected the formation of DAG at the Golgi apparatus. As a consequence, down-regulation of SMS1 and SMS2 r...

  15. Inhibition of sphingomyelin synthase (SMS) affects intracellular sphingomyelin accumulation and plasma membrane lipid organization

    Li, Zhiqiang; Hailemariam, Tiruneh K.; Zhou, Hongwen; Li, Yan; Duckworth, Dale C.; Peake, David A.; Zhang, Youyan; Kuo, Ming-Shang; Cao, Guoqing; Jiang, Xian-Cheng


    Sphingomyelin plays a very important role both in cell membrane formation that may well have an impact on the development of diseases like atherosclerosis and diabetes. However, the molecular mechanism that governs intracellular and plasma membrane SM levels is largely unknown. Recently, two isoforms of sphingomyelin synthase (SMS1 and SMS2), the last enzyme for SM de novo synthesis, have been cloned. We have hypothesized that SMS1 and SMS2 are the two most likely candidates responsible for t...

  16. Subcellular Targeting Domains of Sphingomyelin Synthase 1 and 2

    Yeang Calvin; Ding Tingbo; Chirico William J; Jiang Xian-Cheng


    Abstract Sphingomyelin synthase (SMS) sits at the crossroads of sphingomyelin (SM), ceramide, diacylglycerol (DAG) metabolism. It utilizes ceramide and phosphatidylcholine as substrates to produce SM and DAG, thereby regulating lipid messengers which play a role in cell survival and apoptosis. Furthermore, its product SM has been implicated in atherogenic processes such as retention of lipoproteins in the blood vessel intima. There are two mammalian sphingomyelin synthases: SMS1 and SMS2. SMS...

  17. Cholesterol interactions with ceramide and sphingomyelin.

    García-Arribas, Aritz B; Alonso, Alicia; Goñi, Felix M


    Sphingolipids contain in their polar heads chemical groups allowing them to establish a complex network of H-bonds (through different OH and NHgroups) with other lipids in the bilayer. In the recent years the specific interaction of sphingomyelin (SM) with cholesterol (Chol) has been examined, largely in the context of the "lipid raft" hypothesis. Formation of SM-Ceramide (Cer) complexes, proposed to exist in cell membranes in response to stress, has also been described. More recently, a delicate balance of phase formation and transformation in ternary mixtures of SM, Chol and Cer, with mutual displacement of Chol and Cer from their interaction with SM is considered to exist. In addition, data demonstrating direct Chol-Cer interaction are becoming available. PMID:27132117

  18. Alpha Particles Induce Apoptosis through the Sphingomyelin Pathway

    Seideman, Jonathan H.; Stancevic, Branka; Rotolo, Jimmy A.; McDevitt, Michael R.; Howell, Roger W.; Kolesnick, Richard N; Scheinberg, David A.


    The sphingomyelin pathway involves the enzymatic cleavage of sphingomyelin to produce ceramide, a second messenger that serves as a key mediator in the rapid apoptotic response to various cell stressors. Low-linear energy transfer (LET) γ radiation can initiate this pathway, independent of DNA damage, via the cell membrane. Whether short-ranged, high-LET a particles, which are of interest as potent environmental carcinogens, radiotherapies and potential components of dirty bombs, can act thro...

  19. The Domain Responsible for Sphingomyelin Synthase (SMS) Activity

    Yeang, Calvin; Varsheny, Shweta; Wang, Renxiao; ZHANG, YA; Ye, Deyong; Jiang, Xian-Cheng


    Sphingomyelin synthase (SMS) sits at the crossroads of sphingomyelin (SM), ceramide, diacylglycerol (DAG) metabolism. It utilizes ceramide and phosphatidylcholine as substrates to produce SM and DAG, thereby regulating lipid messengers which play a role in cell survival and apoptosis. There are two isoforms of the enzyme, SMS1 and SMS2. Both SMS1 and SMS2 contain two histidines and one aspartic acid which are evolutionary conserved within the lipid phosphate phosphatase superfamily. In this s...

  20. Excess sphingomyelin disturbs ATG9A trafficking and autophagosome closure.

    Corcelle-Termeau, Elisabeth; Vindeløv, Signe Diness; Hämälistö, Saara; Mograbi, Baharia; Keldsbo, Anne; Bräsen, Jan Hinrich; Favaro, Elena; Adam, Dieter; Szyniarowski, Piotr; Hofman, Paul; Krautwald, Stefan; Farkas, Thomas; Petersen, Nikolaj H T; Rohde, Mikkel; Linkermann, Andreas; Jäättelä, Marja


    Sphingomyelin is an essential cellular lipid that traffics between plasma membrane and intracellular organelles until directed to lysosomes for SMPD1 (sphingomyelin phosphodiesterase 1)-mediated degradation. Inactivating mutations in the SMPD1 gene result in Niemann-Pick diseases type A and B characterized by sphingomyelin accumulation and severely disturbed tissue homeostasis. Here, we report that sphingomyelin overload disturbs the maturation and closure of autophagic membranes. Niemann-Pick type A patient fibroblasts and SMPD1-depleted cancer cells accumulate elongated and unclosed autophagic membranes as well as abnormally swollen autophagosomes in the absence of normal autophagosomes and autolysosomes. The immature autophagic membranes are rich in WIPI2, ATG16L1 and MAP1LC3B but display reduced association with ATG9A. Contrary to its normal trafficking between plasma membrane, intracellular organelles and autophagic membranes, ATG9A concentrates in transferrin receptor-positive juxtanuclear recycling endosomes in SMPD1-deficient cells. Supporting a causative role for ATG9A mistrafficking in the autophagy defect observed in SMPD1-deficient cells, ectopic ATG9A effectively reverts this phenotype. Exogenous C12-sphingomyelin induces a similar juxtanuclear accumulation of ATG9A and subsequent defect in the maturation of autophagic membranes in healthy cells while the main sphingomyelin metabolite, ceramide, fails to revert the autophagy defective phenotype in SMPD1-deficient cells. Juxtanuclear accumulation of ATG9A and defective autophagy are also evident in tissues of smpd1-deficient mice with a subsequent inability to cope with kidney ischemia-reperfusion stress. These data reveal sphingomyelin as an important regulator of ATG9A trafficking and maturation of early autophagic membranes. PMID:27070082

  1. Identification of a family of animal sphingomyelin synthases

    Huitema, K.R.; van den Dikkenberg, J.; Brouwers, J.F.H.M.; Holthuis, J.C.M.


    Sphingomyelin (SM) is a major component of animal plasma membranes. Its production involves the transfer of phosphocholine from phosphatidylcholine onto ceramide, yielding diacylglycerol as a side product. This reaction is catalysed by SM synthase, an enzyme whose biological potential can be judged from the roles of diacylglycerol and ceramide as anti- and proapoptotic stimuli, respectively. SM synthesis occurs in the lumen of the Golgi as well as on the cell surface. As no gene for SM syntha...

  2. Nuclear Phosphatidylcholine and Sphingomyelin Metabolism of Thyroid Cells Changes during Stratospheric Balloon Flight

    Elisabetta Albi


    Full Text Available Nuclear sphingomyelin and phosphatidylcholine metabolism is involved in the response to ultraviolet radiation treatment in different ways related to the physiological state of cells. To evaluate the effects of low levels of radiation from the stratosphere on thyroid cells, proliferating and quiescent FRTL-5 cells were flown in a stratospheric balloon (BIRBA mission. After recovery, the activity of neutral sphingomyelinase, phosphatidylcholine-specific phospholipase C, sphingomyelin synthase, and reverse sphingomyelin synthase was assayed in purified nuclei and the nuclei-free fraction. In proliferating FRTL-5, space radiation stimulate nuclear neutral sphingomyelinase and reverse sphingomyelin synthase activity, whereas phosphatidylcholine-specific phospholipase C and sphingomyelin synthase were inhibited, thus inducing sphingomyelin degradation and phosphatidylcholine synthesis. This effect was lower in quiescent cells. The possible role of nuclear lipid metabolism in the thyroid damage induced by space radiations is discussed.

  3. Detectors for evaluating the cellular landscape of sphingomyelin- and cholesterol-rich membrane domains.

    Kishimoto, Takuma; Ishitsuka, Reiko; Kobayashi, Toshihide


    Although sphingomyelin and cholesterol are major lipids of mammalian cells, the detailed distribution of these lipids in cellular membranes remains still obscure. However, the recent development of protein probes that specifically bind sphingomyelin and/or cholesterol provides new information about the landscape of the lipid domains that are enriched with sphingomyelin or cholesterol or both. Here, we critically summarize the tools to study distribution and dynamics of sphingomyelin and cholesterol. This article is part of a Special Issue entitled: The cellular lipid landscape edited by Tim P. Levine and Anant K. Menon. PMID:26993577

  4. Modulation of human type II secretory phospholipase A2 by sphingomyelin and annexin VI.

    Koumanov, K; Wolf, C; Béreziat, G


    Conjectural results have been reported on the capacity of inflammatory secreted phospholipase A2 (sPLA2) to hydrolyse mammalian membrane phospholipids. Development of an assay based on the release of non-esterified fatty acids by the enzyme acting on the organized phospholipid mixture constituting the membrane matrix has led to the identification of two prominent effectors, sphingomyelin (SPH) and annexin. Recombinant human type II sPLA2 hydrolyses red-cell membrane phospholipids with a marked preference for the inner leaflet. This preference is apparently related to the high content of SPH in the outer leaflet, which inhibits sPLA2. This inhibition by SPH is specific for sPLA2. Cholesterol counteracts the inhibition of sPLA2 by SPH, suggesting that the SPH-to-cholesterol ratio accounts in vivo for the variable susceptibility of cell membranes to sPLA2. Different effects were observed of the presence of the non-hydrolysable D-alpha-dipalmitoyl phosphatidylcholine (D-DPPC), which renders the membranes rigid but does not inhibit sPLA2. Annexin VI was shown, along with other annexins, to inhibit sPLA2 activity by sequestering the phospholipid substrate. The present study has provided the first evidence that annexin VI, in concentrations that inhibit hydrolysis of purified phospholipid substrates, stimulated the hydrolysis of membrane phospholipids by sPLA2. The activation requires the presence of membrane proteins. The effect is specific for type II sPLA2 and is not reproducible with type I PLA2. The activation by annexin VI of sPLA2 acting on red cell membranes results in the preferential release of polyunsaturated fatty acids. It suggests that type II sPLA2, in conjunction with annexin VI, might be involved in the final step of endocytosis and/or exocytosis providing the free polyunsaturated fatty acids acting synergistically to cause membrane fusion. PMID:9337873

  5. Regulation of Cell Migration by Sphingomyelin Synthases: Sphingomyelin in Lipid Rafts Decreases Responsiveness to Signaling by the CXCL12/CXCR4 Pathway

    Asano, Satoshi; Kitatani, Kazuyuki; Taniguchi, Makoto; Hashimoto, Mayumi; Zama, Kota; Mitsutake, Susumu; Igarashi, Yasuyuki; Takeya, Hiroyuki; KIGAWA, JUNZO; Hayashi, Akira; Umehara, Hisanori; Okazaki, Toshiro


    Sphingomyelin synthase (SMS) catalyzes the formation of sphingomyelin, a major component of the plasma membrane and lipid rafts. To investigate the role of SMS in cell signaling and migration induced by binding of the chemokine CXCL12 to CXCR4, we used mouse embryonic fibroblasts deficient in SMS1 and/or SMS2 and examined the effects of SMS deficiency on cell migration. SMS deficiency promoted cell migration through a CXCL12/CXCR4-dependent signaling pathway involving extracellular signal-reg...

  6. Hydrolysis of uranium hexafluoride

    A literature survey is presented of uranium hexafluoride hydrolysis methods as the first step in UF6 conversion to UO2. Reviewed are early methods of hydrolysis, the hydrolysis by dry water vapour, the fluidized-bed method, and the liquid phase hydrolysis of UF6 gas. (J.P.)

  7. The Role of Sphingomyelin Breakdown in Measles Virus Immunmodulation

    Elita Avota


    Full Text Available Measles virus (MV efficiently causes generalized immunosuppression which accounts to a major extent for cases of measles-asscociated severe morbidity and mortality. MV infections alter many functions of antigen presenting cells (APC (dendritic cells (DCs and lymphocytes, yet many molecular targets of the virus remain poorly defined. Cellular interactions and effector functions of DCs and lymphocytes are regulated by surface receptors. Associating with other proteins involved in cell signaling, receptors form part of receptosomes that respond to and transmit external signals through dynamic interctions with the cytoskeleton. Alterations in the composition and metabolism of membrane sphingolipids have a substantial impact on both processes. In this review we focus on the regulation of sphingomyelinase activity and ceramide release in cells exposed to MV and discuss the immunosuppressive role of sphingomyelin breakdown induced by MV.

  8. Nerve growth factor, sphingomyelins, and sensitization in sensory neurons

    Grant D. Nicol


    @@ Because nerve growth factor (NGF) is elevated during inflammation, plays a causal role in the initiation of hyperalgesia, and is known to activate the sphingomyelin signalling pathway, we examined whether NGF and its putative second messenger, ceramide, could modulate the excitability of capsaicin-sensitive adult sensory neurons.Using the whole-cell patch-clamp recording technique,exposure of isolated sensory neurons to either 100 ng/mL NGF or 1 mmol/L N-acetyl sphingosine (C2-ceramide) produced a 3-4 fold increase in the number of action po-tentials (APs) evoked by a ramp of depolarizing current in a time-dependent manner. Intracellular perfusion with bac- terial sphingomyelinase (SMase) also increased the num- ber of APs suggesting that the release of native ceramide enhanced neuronal excitability.

  9. [Activity of the sphingomyelin cycle enzymes and concentration of products of sphingomyelin degradation in the rat liver in the course of acute toxic hepatitis].

    Serebrov, V Iu; Kuz'menko, D I; Burov, P G; Sapugol'tseva, O B


    Activity of key enzymes of a sphingomyelin cycle and the maintenance of its components (sphingomyelin, ceramide and sphingosine-1-phosphate) have been studied in livers of rats in dynamics of the acute toxic hepatitis caused by hypodermic introduction of an oil solution of CCl4. Sphingomyelinase activity significally increased already on early terms and remained increased over the whole period of observation. Activity of ceramidase insignificantly differed from the control level. The levels of sphingomyelin and sphingosine-1-phosphate did not undergo marked changes while ceramide content significally increased. Thus, balance between liver content of ceramide (proapoptotic) and the sphingosine-1-phosphate, being the antiapoptotic factor, was shifted towards ceramide. In sphingomyelin molecules there was a significant decrease in the content of fatty acids C18: and C22:2, while in ceramide molecules and sphingosine-1-phosphate only fatty acid C22:2 changed. In spite of significant decrease in content of some unsaturated fatty acids, calculated unsaturation coefficients of the fatty acid component of the sphingomyelin cycle metabolites. Thus, our results together with literature data suggests involvement of ceramide-mediated apoptosis in the pathogenesis of acute toxic hepatitis. Elimination of damaged hepatocytes facilitates realization of repair processes and optimization of cellular community of a liver. PMID:21341516

  10. 21 CFR 862.1455 - Lecithin/sphingomyelin ratio in amniotic fluid test system.


    ... HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES CLINICAL CHEMISTRY AND CLINICAL TOXICOLOGY DEVICES Clinical Chemistry Test Systems § 862.1455 Lecithin/sphingomyelin ratio in amniotic fluid test system....

  11. Nuclear Phosphatidylcholine and Sphingomyelin Metabolism of Thyroid Cells Changes during Stratospheric Balloon Flight

    Giuseppina Perrella; Maristella Villani; Samuela Cataldi; Elisabetta Albi


    Nuclear sphingomyelin and phosphatidylcholine metabolism is involved in the response to ultraviolet radiation treatment in different ways related to the physiological state of cells. To evaluate the effects of low levels of radiation from the stratosphere on thyroid cells, proliferating and quiescent FRTL-5 cells were flown in a stratospheric balloon (BIRBA mission). After recovery, the activity of neutral sphingomyelinase, phosphatidylcholine-specific phospholipase C, sphingomyelin synthase,...

  12. All members in the sphingomyelin synthase gene family have ceramide phosphoethanolamine synthase activity[S

    Ding, Tingbo; Kabir, Inamul; Li, Yue; Lou, Caixia; Yazdanyar, Amirfarbod; Xu, Jiachen; Dong, Jibin; Zhou, Hongwen; Park, Taesik; Boutjdir, Mohamed; Li, Zhiqiang; Jiang, Xian-Cheng


    Sphingomyelin synthase-related protein (SMSr) synthesizes the sphingomyelin analog ceramide phosphoethanolamine (CPE) in cells. Previous cell studies indicated that SMSr is involved in ceramide homeostasis and is crucial for cell function. To further examine SMSr function in vivo, we generated Smsr KO mice that were fertile and had no obvious phenotypic alterations. Quantitative MS analyses of plasma, liver, and macrophages from the KO mice revealed only marginal changes in CPE and ceramide a...

  13. Sphingomyelin synthase 2 (SMS2) deficiency attenuates LPS-induced lung injury

    Gowda, Satish; Yeang, Calvin; Wadgaonkar, Sunil; Anjum, Fatima; Grinkina, Natalia; Cutaia, Michael; Jiang, Xian-Chen; Wadgaonkar, Raj


    Sphingomyelin synthase (SMS) catalyzes the synthesis of sphingomyelin (SM) and is required for maintenance of plasma membrane microdomain fluidity. Of the two isoforms of mammalian SMS, SMS1 is mostly present in the trans-Golgi apparatus, whereas SMS2 is predominantly found at the plasma membrane. SMS2 has a role in receptor mediated response to inflammation in macrophages, however, the role of SMS2 in vascular permeability, pulmonary edema, and lung injury have not been investigated. To defi...

  14. Sphingomyelin synthase 1 activity is regulated by the BCR-ABL oncogene[S

    Burns, Tara Ann; Subathra, Marimuthu; signorelli, Paola; Choi, Young; Yang, Xiaofeng; Wang, Yong; Villani, Maristella; Bhalla, Kapil; Zhou, Daohong; Luberto, Chiara


    Sphingomyelin synthase (SMS) produces sphingomyelin while consuming ceramide (a negative regulator of cell proliferation) and forming diacylglycerol (DAG) (a mitogenic factor). Therefore, enhanced SMS activity could favor cell proliferation. To examine if dysregulated SMS contributes to leukemogenesis, we measured SMS activity in several leukemic cell lines and found that it is highly elevated in K562 chronic myelogenous leukemia (CML) cells. The increased SMS in K562 cells was caused by the ...

  15. Acylamide elimination during radiation-induced free-radical fragmentation of sphingomyelin

    Radiolysis of sphingomyelin contained in bilayer membranes was studied. Irradiation was realized using γ-facility at a dose rate of 0.35 Gy/s. It is shown that free radical fragmentation of sphingomyelin part of lipid molecule is the most probable process, resulting in spheroylamide formation. It is pointed out that the established way of fragmentation can make a significant contribution to the cell damage under the action of ionizing radiation

  16. Increased Oxidative Stress Impairs Adipose Tissue Function in Sphingomyelin Synthase 1 Null Mice

    Masato Yano; Tadashi Yamamoto; Naotaka Nishimura; Tomomi Gotoh; Ken Watanabe; Kazutaka Ikeda; Yohei Garan; Ryo Taguchi; Koichi Node; Toshiro Okazaki; Yuichi Oike


    Sphingomyelin synthase 1 (SMS1) catalyzes the conversion of ceramide to sphingomyelin. Here, we found that SMS1 null mice showed lipodystrophic phenotype. Mutant mice showed up-regulation of plasma triglyceride concentrations accompanied by reduction of white adipose tissue (WAT) as they aged. Lipoprotein lipase (LPL) activity was severely reduced in mutant mice. In vivo analysis indicated that fatty acid uptake in WAT but not in liver decreased in SMS1 null compared to wild-type mice. In vit...

  17. Interaction of Egg-Sphingomyelin with DOPC in Langmuir Monolayers

    Chang-chun Hao; Run-guang Sun; Jing Zhang


    Lipid rafts are a dynamic microdomain structure found in recent years,enriched in sphingolipids,cholesterol and particular proteins.The change of structure and function of lipid rafts could result in many diseases.In this work,the monolayer miscibility behavior of mixed systems of Egg-Sphingomyelin (ESM) with 1,2-dioleoyl-sn-glycero-3-phosphocholine was investigated in terms of mean surface area per molecule and excess molecular area △Aex at certain surface pressure,surface pressure and excess surface pressure △πex at certain mean molecular area.The stability and compressibility of the mixed monolayers was assessed by the parameters of surface excess Gibbs free energy △Gex,excess Helmholtz energy △Hex and elasticity.Thermodynamic analysis indicates △Aex and △πex in the binary systems with positive deviations from the ideal behavior,suggesting repulsive interaction.The maximum of △Gex and △Hex was at the molar fraction of ESM of 0.6,demonstrating the mixed monolayer was more unstable.The repulsive interaction induced phase separation in the monolayer.

  18. Is there a sphingomyelin-based hydrogen bond barrier at the mammalian host-schistosome parasite interface?

    Migliardo, Federica; Tallima, Hatem; El Ridi, Rashika


    Schistosomes develop, mature, copulate, lay eggs, and live for years in the mammalian host bloodstream, importing nutrients across the tegument, but entirely impervious to the surrounding elements of the immune system. We have hypothesized that sphingomyelin (SM) in the parasite apical lipid bilayer is responsible for these sieving properties via formation of a tight hydrogen bond network with the surrounding water. Here we have used quasi-elastic neutron scattering for characterizing the diffusion of larval and adult Schistosoma mansoni and adult Schistosoma haematobium in the surrounding medium, under various environmental conditions. The results documented the presence of a hydrogen bond barrier around larvae and adult schistosomes. The hydrogen bond network readily collapses if worms are subjected to hypoxic conditions, likely via activation of the parasite tegument-associated neutral sphingomyelinase, and consequent excessive SM hydrolysis. The slower dynamics of lung-stage larvae as compared to adult worms has been related to the existence of hydrogen-bonded networks of different strength and then to their differential resistance to immune attacks. PMID:23943053

  19. Nuclear Lipid Microdomain as Place of Interaction between Sphingomyelin and DNA during Liver Regeneration

    Samuela Cataldi


    Full Text Available Nuclear sphingomyelin is a key molecule for cell proliferation. This molecule is organized with cholesterol and proteins to form specific lipid microdomains bound to the inner nuclear membrane where RNA is synthesized. Here, we have reported the ability of the sphingomyelin present in the nuclear microdomain to bind DNA and regulate its synthesis, and to highlight its role in cell proliferation induced by partial hepatectomy. During G1/S transition of the cell cycle, sphingomyelin and DNA content is very high and it is strongly reduced after exogenous sphingomyelinase treatment. During the S-phase of the cell cycle, the stimulation of sphingomyelinase and inhibition of sphingomyelin–synthase are accompanied by the DNA synthesis start. To assess the specificity of the results, experiments were repeated with trifluoperazine, a drug known to affect the synthesis of lipids and DNA and to stimulate sphingomyelinase activity. The activity of sphingomyelinase is stimulated in the first hour after hepatectomy and sphingomyelin–DNA synthesis is strongly attenuated. It may be hypothesized that the nuclear microdomain represents a specific area of the inner nuclear membrane that acts as an active site of chromatin anchorage thanks to the stabilizing action of sphingomyelin. Thus, sphingomyelin metabolism in nuclear lipid microdomains is suggested to regulate cell proliferation.

  20. Both Sphingomyelin Synthases SMS1 and SMS2 Are Required for Sphingomyelin Homeostasis and Growth in Human HeLa Cells

    Geta Tafesse, F.; Huitema, K.R.; Hermansson, M; van der Poel, S.; van den Dikkenberg, J.; Uphoff, A.; Somerharju, P; Holthuis, J.C.M.


    Sphingomyelin (SM) is a vital component of cellular membranes in organisms ranging from mammals to protozoa. Its production involves the transfer of phosphocholine from phosphatidylcholine to ceramide, yielding diacylglycerol in the process. The mammalian genome encodes two known SM synthase (SMS) isoforms, SMS1 and SMS2. However, the relative contributions of these enzymes to SM production in mammalian cells remained to be established. Here we show that SMS1 and SMS2 are co-expressed in a va...

  1. Crucial role of alkaline sphingomyelinase in sphingomyelin digestion: a study on enzyme knockout mice

    Zhang, Yao; Cheng, Yajun; Hansen, Gert H;


    Alkaline sphingomyelinase (alk-SMase) hydrolyses sphingomyelin (SM) to ceramide in the gut. To evaluate the physiological importance of the enzyme, we generated alk-SMase knockout (KO) mice by the Cre-recombinase-Locus of X-over P1(Cre-LoxP) system and studied SM digestion. Both wild-type (WT...

  2. Functional activity of sphingomyelin cycle in rat liver in chronic toxic hepatitis.

    Serebrov, V Yu; Kuzmenko, D I; Burov, P G; Novitsky, S V


    Activities of sphingomyelinase and ceramidase decreased in the liver in chronic toxic hepatitis and the balance between the levels of proapoptotic ceramide and antiapoptotic sphyngosine-1-phosphate shifts towards the latter substance. Pronounced changes in the qualitative and quantitative composition of fatty acids in the sphingomyelin cycle effector molecules were revealed. PMID:19513367

  3. A Lipidomic Study of the Effects of N-methyl-N'-nitro-N-nitrosoguanidine on Sphingomyelin Metabolism

    Yun HUANG; Jing SHEN; Ting WANG; Yan-Ke YU; Fanqing F. CHEN; Jun YANG


    Systems biology is a new and rapidly developing research area in which, by quantitatively describing the interaction among all the individual components of a cell, a systems-level understanding of a biological response can be achieved. Therefore, it requires high-throughput measurement technologies for biological molecules, such as genomic and proteomic approaches for DNA/RNA and protein, respectively.Recently, a new concept, lipidomics, which utilizes the mass spectrometry (MS) method for lipid analysis,has been proposed. Using this lipidomic approach, the effects ofN-methyl-N'-nitro-N-nitrosoguanidine (MNNG) on sphingomyelin metabolism, a major class of sphingolipids, were evaluated. Sphingomyelin molecules were extracted from cells and analyzed by matrix-assisted laser desorption ionization-time of flight MS. It was found that MNNG induced profound changes in sphingomyelin metabolism, including the appearance of some new sphingomyelin species and the disappearance of some others, and the concentrations of several sphingomyelin species also changed. This was accompanied by the redistribution of acid sphingomyelinase (ASM), a key player in sphingomyelin metabolism. On the other hand, imipramine, an inhibitor of ASM, caused the accumulation of sphingomyelin. It also prevented some of the effects of MNNG, as well as the redistribution of ASM. Taken together, these data suggested that the lipidomic approach is highly effective for the systematic analysis of cellular lipids metabolism.

  4. Dynamic Modification of Sphingomyelin in Lipid Microdomains Controls Development of Obesity, Fatty Liver, and Type 2 Diabetes*

    Mitsutake, Susumu; Zama, Kota; Yokota, Hazuki; Yoshida, Tetsuya; Tanaka, Miki; Mitsui, Masaru; Ikawa, Masahito; Okabe, Masaru; Tanaka, Yoshikazu; Yamashita, Tadashi; Takemoto, Hiroshi; Okazaki, Toshiro; Watanabe, Ken; Igarashi, Yasuyuki


    Lipid microdomains or caveolae, small invaginations of plasma membrane, have emerged as important elements for lipid uptake and glucose homeostasis. Sphingomyelin (SM) is one of the major phospholipids of the lipid microdomains. In this study, we investigated the physiological function of sphingomyelin synthase 2 (SMS2) using SMS2 knock-out mice, and we found that SMS2 deficiency prevents high fat diet-induced obesity and insulin resistance. Interestingly, in the liver of SMS2 knock-out mice,...

  5. Detailed Comparison of Deuterium Quadrupole Profiles between Sphingomyelin and Phosphatidylcholine Bilayers

    Yasuda, Tomokazu; Kinoshita, Masanao; Murata, Michio; Matsumori, Nobuaki


    Lipid rafts are microdomains rich in sphingomyelin (SM) and cholesterol (Chol). The essential question is why natural lipid rafts prefer SM rather than saturated diacyl glycerophosphocholine, although both form ordered membranes with Chol in model systems. Hence in this study, we synthesized site-specifically deuterated 1-palmitoyl-2-stearoyl-sn-glycero-3-phosphocholines that match the acyl chain length of stearoyl-SM (SSM), and compared their deuterium quadrupole coupling profiles in detail....

  6. The Chlamydial Inclusion Preferentially Intercepts Basolaterally Directed Sphingomyelin-Containing Exocytic Vacuoles

    Moore, Elizabeth R.; Fischer, Elizabeth R.; Mead, David J.; Hackstadt, Ted


    Chlamydiae replicate intracellularly within a unique vacuole termed the inclusion. The inclusion circumvents classical endosomal/lysosomal pathways but actively intercepts a subset of Golgi-derived exocytic vesicles containing sphingomyelin (SM) and cholesterol. To further examine this interaction, we developed a polarized epithelial cell model to study vectoral trafficking of lipids and proteins to the inclusion. We examined seven epithelial cell lines for their ability to form single monola...

  7. Enzymatic Hydrolysis of Lignocelluloses

    Kolasa, Marta; Ahring, Birgitte Kiær; Lübeck, Peter Stephensen;


    bonds. Cellulose can be degraded to simple sugar components by means of enzymatic hydrolysis. However, due to its complex, crystalline structure it is difficult to break it down and the cooperative action of a variety of cellulolytic enzymes is necessary. Fungi are known to have potential in production...... of a variety of cellulolytic enzymes. The aim of this work is to discover new thermostable and robust cellulolytic enzymes for improved enzymatic hydrolysis of biomass. For this purpose two screening methods are applied in different fungal strains with high cellulolytic activities: an expression...

  8. Cholesterol and Sphingomyelin-Containing Model Condensed Lipid Monolayers: Heterogeneities Involving Ordered Microdomains Assessed by Two Cholesterol Derivatives.

    Lecompte, Marie-France; Gaibelet, Gérald; Lebrun, Chantal; Tercé, François; Collet, Xavier; Orlowski, Stéphane


    Lipid monolayers are often considered as model membranes, but they are also the physiologic lipid part of the peripheral envelope of lipoproteins and cytosolic lipid bodies. However, their structural organization is still rather elusive, in particular when both cholesterol and sphingomyelin are present. To investigate such structural organization of hemimembranes, we measured, using alternative current voltammetry, the differential capacitance of condensed phosphatidylcholine-based monolayers as a function of applied potential, which is sensitive to their lipid composition and molecular arrangement. Especially, monolayers containing both sphingomyelin and cholesterol, at 15% w/w, presented specific characteristics of the differential capacitance versus potential curves recorded, which was indicative of specific interactions between these two lipid components. We then compared the behavior of two cholesterol derivatives (at 15% w/w), 21-methylpyrenyl-cholesterol (Pyr-met-Chol) and 22-nitrobenzoxadiazole-cholesterol (NBD-Chol), with that of cholesterol when present in model monolayers. Indeed, these two probes were chosen because of previous findings reporting opposite behaviors within bilayer membranes regarding their interaction with ordered lipids, with only Pyr-met-Chol mimicking cholesterol well. Remarkably, in monolayers containing sphingomyelin or not, Pyr-met-Chol and NBD-Chol presented contrasting behaviors, and Pyr-met-Chol mimicked cholesterol only in the presence of sphingomyelin. These two observations (i.e., optimal amounts of sphingomyelin and cholesterol, and the ability to discriminate between Pyr-met-Chol and NBD-Chol) can be interpreted by the existence of heterogeneities including ordered patches in sphingomyelin- and cholesterol-containing monolayers. Since such monolayer lipid arrangement shares some properties with the raft-type lipid microdomains well-described in sphingomyelin- and cholesterol-containing bilayer membranes, our data thus

  9. Structural Basis for Nucleotide Hydrolysis by the Acid Sphingomyelinase-like Phosphodiesterase SMPDL3A.

    Gorelik, Alexei; Illes, Katalin; Superti-Furga, Giulio; Nagar, Bhushan


    Sphingomyelin phosphodiesterase, acid-like 3A (SMPDL3A) is a member of a small family of proteins founded by the well characterized lysosomal enzyme, acid sphingomyelinase (ASMase). ASMase converts sphingomyelin into the signaling lipid, ceramide. It was recently discovered that, in contrast to ASMase, SMPDL3A is inactive against sphingomyelin and, surprisingly, can instead hydrolyze nucleoside diphosphates and triphosphates, which may play a role in purinergic signaling. As none of the ASMase-like proteins has been structurally characterized to date, the molecular basis for their substrate preferences is unknown. Here we report crystal structures of murine SMPDL3A, which represent the first structures of an ASMase-like protein. The catalytic domain consists of a central mixed β-sandwich surrounded by α-helices. Additionally, SMPDL3A possesses a unique C-terminal domain formed from a cluster of four α-helices that appears to distinguish this protein family from other phosphoesterases. We show that SMDPL3A is a di-zinc-dependent enzyme with an active site configuration that suggests a mechanism of phosphodiester hydrolysis by a metal-activated water molecule and protonation of the leaving group by a histidine residue. Co-crystal structures of SMPDL3A with AMP and α,β-methylene ADP (AMPCP) reveal that the substrate binding site accommodates nucleotides by establishing interactions with their base, sugar, and phosphate moieties, with the latter the major contributor to binding affinity. Our study provides the structural basis for SMPDL3A substrate specificity and sheds new light on the function of ASMase-like proteins. PMID:26792860

  10. The role of cholesterol-sphingomyelin membrane nanodomains in the stability of intercellular membrane nanotubes

    Veranič P


    Full Text Available Maruša Lokar1,*, Doron Kabaso1,2,*, Nataša Resnik3, Kristina Sepcic5, Veronika Kralj-Iglic4,6, Peter Veranic3, Robert Zorec2, Aleš Iglic1,6 1Laboratory of Biophysics, Faculty of Electrical Engineering, 2Laboratory of Neuroendocrinology-Molecular Cell Physiology, Faculty of Medicine, 3Institute of Cell Biology, Faculty of Medicine, 4Faculty of Health Sciences, 5Department of Biology, Biotechnical Faculty, 6Laboratory of Clinical Biophysics, Department of Orthopedic Surgery, Faculty of Medicine, University of Ljubljana, Ljubljana, Slovenia*These authors equally share first authorshipAbstract: Intercellular membrane nanotubes (ICNs are highly curved tubular structures that connect neighboring cells. The stability of these structures depends on the inner cytoskeleton and the cell membrane composition. Yet, due to the difficulty in the extraction of ICNs, the cell membrane composition remains elusive. In the present study, a raft marker, ostreolysin, revealed the enrichment of cholesterol-sphingomyelin membrane nanodomains along ICNs in a T24 (malignant urothelial cancer cell line. Cholesterol depletion, due to the addition of methyl-β-cyclodextrin, caused the dispersion of cholesterol-sphingomyelin membrane nanodomains and the retraction of ICNs. The depletion of cholesterol also led to cytoskeleton reorganization and to formation of actin stress fibers. Live cell imaging data revealed the possible functional coupling between the change from polygonal to spherical shape, cell separation, and the disconnection of ICNs. The ICN was modeled as an axisymmetric tubular structure, enabling us to investigate the effects of cholesterol content on the ICN curvature. The removal of cholesterol was predicted to reduce the positive spontaneous curvature of the remaining membrane components, increasing their curvature mismatch with the tube curvature. The mechanisms by which the increased curvature mismatch could contribute to the disconnection of ICNs are

  11. Improved sensitivity of an acid sphingomyelinase activity assay using a C6:0 sphingomyelin substrate

    Wei-Lien Chuang; Joshua Pacheco; Samantha Cooper; Kingsbury, Jonathan S.; John Hinds; Pavlina Wolf; Petra Oliva; Joan Keutzer; Cox, Gerald F.; Kate Zhang


    Short-chain C6-sphingomyelin is an artificial substrate that was used in an acid sphingomyelinase activity assay for a pilot screening study of patients with Niemann–Pick disease types A and B. Using previously published multiplex and single assay conditions, normal acid sphingomyelinase activity levels (i.e. false negative results) were observed in two sisters with Niemann–Pick B who were compound heterozygotes for two missense mutations, p.C92W and p.P184L, in the SMPD1 gene. Increasing the...

  12. X-ray diffraction and calorimetric study of N-lignoceryl sphingomyelin membranes.

    Maulik, P R; Shipley, G G


    Differential scanning calorimetry and x-ray diffraction have been used to investigate hydrated multibilayers of N-lignoceryl sphingomyelin (C24:0-SM) in the hydration range 0-75 wt % H2O. Anhydrous C24:0-SM exhibits a single endothermic transition at 81.3 degrees C (delta H = 3.6 kcal/mol). At low hydration (12.1 wt % H2O), three different endothermic transitions are observed: low-temperature transition (T1) at 39.4 degrees C (transition enthalpy (delta H1) = 2.8 kcal/mol), intermediate-tempe...

  13. Structure of Human Acid Sphingomyelinase Reveals the Role of the Saposin Domain in Activating Substrate Hydrolysis.

    Xiong, Zi-Jian; Huang, Jingjing; Poda, Gennady; Pomès, Régis; Privé, Gilbert G


    Acid sphingomyelinase (ASM) is a lysosomal phosphodiesterase that catalyzes the hydrolysis of sphingomyelin to produce ceramide and phosphocholine. While other lysosomal sphingolipid hydrolases require a saposin activator protein for full activity, the ASM polypeptide incorporates a built-in N-terminal saposin domain and does not require an external activator protein. Here, we report the crystal structure of human ASM and describe the organization of the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain. The saposin domain is tightly associated along an edge of the large, bowl-shaped catalytic domain and adopts an open form that exposes a hydrophobic concave surface approximately 30Å from the catalytic center. The calculated electrostatic potential of the enzyme is electropositive at the acidic pH of the lysosome, consistent with the strict requirement for the presence of acidic lipids in target membranes. Docking studies indicate that sphingomyelin binds with the ceramide-phosphate group positioned at the binuclear zinc center and molecular dynamic simulations indicate that the intrinsic flexibility of the saposin domain is important for monomer-dimer exchange and for membrane interactions. Overall, ASM uses a combination of electrostatic and hydrophobic interactions to cause local disruptions of target bilayers in order to bring the lipid headgroup to the catalytic center in a membrane-bound reaction. PMID:27349982

  14. Hydrolysis of lithium hydride

    Due to its high hydrogen density and unique nuclear chemistry, lithium hydride, in all its isotopic forms, has an unsurpassed place in modem nuclear weapons. The hydrolysis of the material, and the outgassing of hydrogen from the bulk, are crucial to the performance of the material in service. This thesis describes research conducted at AWE Aldermaston, UK, to examine the hydrolysis and hydrogen outgassing from the bulk material, with the aim of ultimately developing the kinetics 8c mechanisms responsible. The basic chemistry is of great interest, especially the reaction with water. This reaction, whilst being fairly extensively studied in the past, has not been conclusively described with an accepted mechanism and associated kinetics. The last significant UK work on the topic was by Imperial College, London, under contract to AW(R)E in the late 1960s. This thesis describes the development of: (i) a solid state NMR spectroscopy technique to examine semi-quantitatively the surface of bulk lithium hydride for its chemical composition, and (ii) a dedicated lithium hydride inert atmosphere gravimetric analysis glove box to study the hydride/water reaction. Solid State NMR Spectroscopy has been utilised for the first time to probe the hydride/hydroxide ratio of partially hydrolysed lithium hydride. 6Li chemical shifts have been established for species of interest and extremely long, up to 17 hours, T1 relaxation times have been measured for 6Li hydride and hydroxide. A method for semi-quantitatively determining the hydroxide/hydride composition of a partially reacted sample has been developed, based on a 'dual-scan' technique using one short and one long pulse sequence. Gravimetric analysis has been developed for lithium hydride/humidity studies. This facility fully contains gravimetric analysis within an argon glove box, with the ability to control the sample atmosphere from room temperature to 60 deg C and from 0.5 to 40 percent relative humidity. The hydrolysis of

  15. Adenovirus-mediated sphingomyelin synthase 2 increases atherosclerotic lesions in ApoE KO mice

    Zhao Yarui


    Full Text Available Abstract Background Sphingomyelin synthase 2 (SMS2 contributes to de novo sphingomyelin (SM biosynthesis. Its activity is related to SM levels in the plasma and the cell membrane. In this study, we investigated the possibility of a direct relationship between SMS and atherosclerosis. Methods The Adenovirus containing SMS2 gene was given into 10-week ApoE KO C57BL/6J mice by femoral intravenous injection. In the control group, the Adenovirus containing GFP was given. To confirm this model, we took both mRNA level examination (RT-PCR and protein level examination (SMS activity assay. Result We generated recombinant adenovirus vectors containing either human SMS2 cDNA (AdV-SMS2 or GFP cDNA (AdV-GFP. On day six after intravenous infusion of 2 × 1011 particle numbers into ten-week-old apoE KO mice, AdV-SMS2 treatment significantly increased liver SMS2 mRNA levels and SMS activity (by 2.7-fold, 2.3-fold, p Conclusions Our results present direct morphological evidence for the pro-atherogenic capabilities of SMS2. SMS2 could be a potential target for treating atherosclerosis.

  16. Isolation of plasma membranes from cultured glioma cells and application to evaluation of membrane sphingomyelin turnover

    A rapid and reliable method for the isolation of plasma membranes and microsomes of high purity and yield from cultured glioma cells is described. The procedure involves disruption by N2 cavitation, preliminary separation by centrifugation in Tricine buffer, and final separation on a gradient formed from 40% Percoll at pH 9.3. Enzyme and chemical markers indicated greater than 60% yield with six- to eightfold enrichment for plasma membranes and greater than 25% yield with three- to fourfold enrichment for a microsomal fraction consisting mainly of endoplasmic reticulum. The final fractions were obtained with high reproducibility in less than 1 h from the time of cell harvesting. Application of this procedure to human fibroblasts in culture is assessed. The isolation procedure was applied to investigations of synthesis and turnover of sphingomyelin and phosphatidylcholine in plasma membranes of glioma cells following incubation for 4-24 h with [methyl-3H]choline. These studies indicated that radioactivity from phosphatidylcholine synthesized in microsomes from exogenous choline may serve as a precursor of the head-group of sphingomyelin accumulating in the plasma membrane

  17. Guanidination of notexin alters its membrane-damaging activity in response to sphingomyelin and cholesterol

    Pei-Hsiu Kao; Yi-Ling Chiou; Shinne-Ren Lin; Long-Sen Chang


    To elucidate the contribution of phospholipase A2 (PLA2) activity of notexin to its ability to perturb membranes, comparative studies on the interaction of notexin and guanidinated notexin (Gu-notexin) with egg yolk phosphatidylcholine (EYPC), EYPC/egg yolk sphingomyelin (EYSM) and EYPC/EYSM/cholesterol vesicles were conducted. EYSM notably reduced the membrane-damaging activity of notexin against EYPC vesicles, but had an insignificant influence on that of Gu-notexin. Unlike the effects noted with notexin, inactivation of PLA2 activity by EDTA led to a reduction in the ability of Gu-notexin to induce EYPC/EYSM vesicle leakage and to increase Gu-notexin-induced membrane permeability of EYPC/EYSM/cholesterol vesicles. The geometrical arrangement of notexin and Gu-notexin in contact with either EYPC/EYSM vesicles or EYPC/EYSM/cholesterol vesicles differed. Moreover, global conformation of notexin and Gu-notexin differed in either Ca2+-bound or metal-free states. These results indicate that notexin and Gu-notexin could induce membrane permeability without the involvement of PLA2 activity, and suggest that guanidination alters the membrane-bound mode of notexin on damaging phospholipid vesicles containing sphingomyelin and cholesterol.

  18. Enzymatic hydrolysis of polyester fabrics

    Enzymatic hydrolysis of polyester fabrics has been investigated, using different treatment times, temperature and concentration of enzymes. The effects of hydrolysis on samples were evaluated by measurement of weight loss, moisture regain, breaking load of warp yarns, thickness and Ftir spectroscopy. Results show that hydrolysis under mild conditions can improve moisture absorption of the samples. If the applied temperature, treatment time and concentration exceeded some specific range, the moisture regain would be affected negatively. The Ftir spectrums showed an increase in functional groups specially hydroxyl. However the effects of enzymatic hydrolysis on weight loss, tensile strength and thickness of polyester fabrics were negligible

  19. Improved sensitivity of an acid sphingomyelinase activity assay using a C6:0 sphingomyelin substrate.

    Chuang, Wei-Lien; Pacheco, Joshua; Cooper, Samantha; Kingsbury, Jonathan S; Hinds, John; Wolf, Pavlina; Oliva, Petra; Keutzer, Joan; Cox, Gerald F; Zhang, Kate


    Short-chain C6-sphingomyelin is an artificial substrate that was used in an acid sphingomyelinase activity assay for a pilot screening study of patients with Niemann-Pick disease types A and B. Using previously published multiplex and single assay conditions, normal acid sphingomyelinase activity levels (i.e. false negative results) were observed in two sisters with Niemann-Pick B who were compound heterozygotes for two missense mutations, p.C92W and p.P184L, in the SMPD1 gene. Increasing the sodium taurocholate detergent concentration in the assay buffer lowered the activity levels of these two patients into the range observed with other patients with clear separation from normal controls. PMID:26937397

  20. Sphingomyelin lipidosis (Niemann-Pick disease) in a juvenile raccoon (Procyon lotor).

    Vapniarsky, N; Wenger, D A; Scheenstra, D; Mete, A


    A wild caught juvenile male raccoon with neurological disease was humanely destroyed due to poor prognosis. Necropsy examination revealed hepatomegaly, splenomegaly and multicentric lymphadenomegaly with diffuse hepatic pallor and pulmonary consolidation with pinpoint pale subpleural foci. Microscopically, there was marked pale cytoplasmic swelling of the central and peripheral neurons as well as the glial cells in the brain, accompanied by multiorgan infiltration by abundant foamy macrophages. Ultrastructural investigation revealed accumulation of concentrically arranged lamellar material within lysosomes of the affected neurons, macrophages and endothelial cells. Biochemical enzymatic analysis detected sphingomyelinase deficiency and lysosomal storage disease consistent with sphingomyelin lipidosis (Niemann-Pick disease [NPD]) was diagnosed. This is the first report of NPD in a raccoon. PMID:23582974

  1. Altered levels of acylcarnitines, phosphatidylcholines, and sphingomyelins in peritoneal fluid from ovarian endometriosis patients.

    Vouk, Katja; Ribič-Pucelj, Martina; Adamski, Jerzy; Rižner, Tea Lanišnik


    Endometriosis is a complex, polygenic, and estrogen-dependent disease that affects 6% to 10% of women of reproductive age, and 30% to 50% of women with infertility and/or pelvic pain. Surgical diagnosis of endometriosis is still the gold standard, as there are currently no diagnostic biomarkers available. Due to the invasive diagnostics, it can take up to 11 years before affected women are diagnosed and receive the appropriate treatment. We performed a targeted metabolomics study to search for potential semi-invasive biomarkers in peritoneal fluid from endometriosis patients. Our case-control study comprised 29 ovarian endometriosis patients and 36 healthy control women. The 148 metabolites included acylcarnitines, glycerophospholipids, and sphingolipids, which were quantified by electrospray ionization tandem mass spectrometry. The strength of association between the metabolites and the metabolite ratios and disease was assessed using crude and adjusted odds ratios. The best combination of biomarkers was then selected by performing step-wise logistic regression. Our analysis reveals significantly decreased concentrations of 10 metabolites, of carnitine and acylcarnitines (C0, C8:1, C6C4:1 DC, C10:1), phosphatidylcholines (PC aa C38:3, PC aa C38:4, PC aa C40:4, PC aa C40:5), and sphingomyelins (SM C16:1, SM C18:1), and 125 significantly altered metabolite ratios in patients versus control women. The best model includes two ratios: a carnitine to a phosphatidylcholine (C0/PC ae C36:0); and between two phosphatidylcholines (PC aa C30:0/PC ae C32:2). When adjusted for age, this provides sensitivity of 82.8% and specificity of 94.4%, with AUC of 0.944. Our study supports the importance of carnitine, phosphatidylcholine, and sphingomyelin metabolites in the pathophysiology of endometriosis, and confirms the potential for the combination of individual metabolite ratios to provide biomarkers for semi-invasive diagnostics. PMID:26921767

  2. [Sphingomyelin synthase 2 deficiency decreases atherosclerosis and inhibits inflammation in mice].

    Qin, Rui; Chen, Ming-Liang; Zhu, Ke; Deng, Jin-Bo; Shi, Yuan-Yuan


    Plasma sphingomyelin (SM) has been shown to be an independent risk factor for coronary heart disease, and sphingomyelin synthase 2 (SMS2) contributes to de novo SM biosynthesis and plasma membrane SM levels. The aim of the present study is to evaluate the in vivo role of SMS2 deficiency in serum SM metabolism and atherosclerosis (AS) development. We used male SMS2 knockout (SMS2(-/-)) and C57BL/6J (wild-type, WT) mice as experimental and control groups, respectively. Each group was fed high-fat diet (1% cholesterol, 20% leaf fat), as well as bile salt for accelerating the atherosclerotic formation. After three months of feeding, the mice were killed to observe aortic arches and oil red-stained longitudinal sections of thoracoabdominal aortae. Fasting blood samples were taken from the tail vein before and after high-fat diet, and the serum lipid and SM levels were measured by using kits and enzymatic method respectively. Western blot was used to analyze the contents of nuclear factor-kappaB (NFkappaB) p65 subunit in peritoneal macrophages stimulated with lipopolysaccharide (LPS) after high-fat diet. The results showed that after high-fat diet, SMS2(-/-) mice presented decreased atherosclerotic lesions in aortic arch and thoracoabdominal aorta compared with WT mice. Regardless of whether high-fat diet were given or not, SMS2(-/-) mice showed a significant decrease in serum SM level (Pstimulation compared with those of the WT mice. These results suggest that SMS2 deficiency decreases AS and inhibits inflammation in mice. Thus, SMS2 deficiency may be a potential therapeutic strategy. PMID:20717634

  3. Effects of chronic injection of sphingomyelin-containing liposomes on lymphoid and non-lymphoid cells in the spleen. Transient suppression of marginal zone macrophages.

    E. Claassen; Westerhof, Y.; Versluis, B.; Kors, N.; Schellekens, M.; Van Rooijen, N.


    Mice were injected with sphingomyelin/cholesterol or phosphatidylcholine/cholesterol (PC/C) liposomes, from twice up to 10 times, on alternate days. Administration of sphingomyelin/cholesterol (SM/C) liposomes gave rise to hepato and splenomegaly, microgranulomatous infections and changes in macrophage numbers and activity in spleen and liver. Enzyme and immuno-cytochemical methods were used, to demonstrate the effect of liposomes on the lymphoid and non-lymphoid cell populations, on cryostat...

  4. Suppression of sphingomyelin synthase 1 by small interference RNA is associated with enhanced ceramide production and apoptosis after photodamage

    Separovic, Duska; Semaan, Louie; Tarca, Adi L.; Maitah, Ma’In Yehya Awad; Hanada, Kentaro; Bielawski, Jacek; Villani, Maristella; Luberto, Chiara


    We have shown that overexpression of SMS1, an enzyme that converts de novo ceramide into sphingomyelin, is accompanied by attenuated ceramide response and apoptotic resistance after photodamage with the photosensitizer Pc 4 (photodynamic therapy; PDT). To test whether SMS1 overexpression-related effects after PDT can be reversed, in this study SMS1 was downregulated in Jurkat T lymphoma/leukemia cells using small inhibitory RNA (siRNA) for SMS1. Compared to scrambled (control) siRNA-transfect...

  5. Sphingomyelin-cholesterol liposomes significantly enhance the pharmacokinetic and therapeutic properties of vincristine in murine and human tumour models.

    Webb, M S; Harasym, T. O.; Masin, D.; Bally, M. B.; Mayer, L. D.


    This study reports on the development of a liposomal formulation of vincristine with significantly enhanced stability and biological properties. The in vitro and in vivo pharmacokinetic, tumour delivery and efficacy properties of liposomal vincristine formulations based on sphingomyelin (SM) and cholesterol were compared with liposomes composed of distearoylphosphatidylcholine (DSPC) and cholesterol. SM/cholesterol liposomes had significantly greater in vitro stability than did similar DSPC/c...

  6. Detailed comparison of deuterium quadrupole profiles between sphingomyelin and phosphatidylcholine bilayers.

    Yasuda, Tomokazu; Kinoshita, Masanao; Murata, Michio; Matsumori, Nobuaki


    Lipid rafts are microdomains rich in sphingomyelin (SM) and cholesterol (Chol). The essential question is why natural lipid rafts prefer SM rather than saturated diacyl glycerophosphocholine, although both form ordered membranes with Chol in model systems. Hence in this study, we synthesized site-specifically deuterated 1-palmitoyl-2-stearoyl-sn-glycero-3-phosphocholines that match the acyl chain length of stearoyl-SM (SSM), and compared their deuterium quadrupole coupling profiles in detail. The results suggest a deeper distribution of Chol in the SSM membranes, a lower entropic penalty upon accommodation of Chol in SSM membranes, and a higher thermal stability of acyl-chain orders in the SSM-Chol bilayers than in the 1-palmitoyl-2-stearoyl-sn-glycero-3-phosphocholine-Chol system at various Chol concentrations. The entropy effect and thermal stability should render SM a more preferred raft constituent than saturated diacyl glycerophosphocholine. Our data also demonstrate that the selective and comprehensive deuteration strategy is indispensable for accurate comparison of order profiles. PMID:24507603

  7. Sphingomyelin in High-Density Lipoproteins: Structural Role and Biological Function

    Jesús Osada


    Full Text Available High-density lipoprotein (HDL levels are an inverse risk factor for cardiovascular diseases, and sphingomyelin (SM is the second most abundant phospholipid component and the major sphingolipid in HDL. Considering the marked presence of SM, the present review has focused on the current knowledge about this phospholipid by addressing its variable distribution among HDL lipoparticles, how they acquire this phospholipid, and the important role that SM plays in regulating their fluidity and cholesterol efflux from different cells. In addition, plasma enzymes involved in HDL metabolism such as lecithin–cholesterol acyltransferase or phospholipid transfer protein are inhibited by HDL SM content. Likewise, HDL SM levels are influenced by dietary maneuvers (source of protein or fat, drugs (statins or diuretics and modified in diseases such as diabetes, renal failure or Niemann–Pick disease. Furthermore, increased levels of HDL SM have been shown to be an inverse risk factor for coronary heart disease. The complexity of SM species, described using new lipidomic methodologies, and their distribution in different HDL particles under many experimental conditions are promising avenues for further research in the future.

  8. Modulation of lipoprotein metabolism by inhibition of sphingomyelin synthesis in ApoE knockout mice.

    Park, Tae-Sik; Panek, Robert L; Rekhter, Mark D; Mueller, Sandra Bak; Rosebury, Wendy S; Robertson, Andrew; Hanselman, Jeffrey C; Kindt, Erick; Homan, Reynold; Karathanasis, Sotirios K


    Plasma sphingomyelin (SM) has been suggested as a risk factor for coronary heart disease independent of cholesterol levels. A decrease of SM in lipoproteins is known to improve the activities of lecithin:cholesterol acyltransferase (LCAT) and lipoprotein lipase (LPL) in vitro. Inhibition of SM biosynthesis may reduce lipoprotein SM content and thus improve cholesterol distribution in lipoproteins by enhancing reverse cholesterol transport and clearance of triglyceride-rich lipoproteins. To examine this hypothesis, ApoE KO mice were fed a western diet and treated for 4 weeks with various concentrations of myriocin, a specific inhibitor of serine palmitoyltransferase. Myriocin treatment lowered plasma cholesterol and TG levels in a dose-dependent manner. In addition, myriocin treatment reduced cholesterol contents in VLDL and LDL and elevated HDL-cholesterol. Observed lipid-lowering effects of myriocin were associated with suppression of HMG CoA reductase and fatty acid synthase via reduced levels of SREBP-1 RNA and protein. Induction of apoAI and lecithin:cholesterol acytransferase (LCAT) in the liver by myriocin was associated with an increased HDL. Lesion area and macrophage area were also diminished in the cuffed femoral artery of ApoE KO mice. In conclusion, inhibition of sphingolipid biosynthesis can be a novel therapeutic target for dyslipidemia and atherosclerosis. PMID:16458317

  9. An enzyme combination assay for serum sphingomyelin: Improved specificity through avoiding the interference with lysophosphatidylcholine.

    Kimura, Takehide; Kuwata, Hideyuki; Miyauchi, Kazuhito; Katayama, Yuki; Kayahara, Norihiko; Sugiuchi, Hiroyuki; Matsushima, Kazumi; Kondo, Yuki; Ishitsuka, Yoichi; Irikura, Mitsuru; Irie, Tetsumi


    Serum sphingomyelin (SM) has predictive value in the development of atherosclerosis. Furthermore, SM plays important roles in cell membrane structure, signal transduction pathways, and lipid raft formation. A convenient enzymatic method for SM is available for routine laboratory practice, but the enzyme specificity is not sufficient because of nonspecific reactions with lysophosphatidylcholine (LPC). Based on the differential specificity of selected enzymes toward choline-containing phospholipids, a two-step assay for measuring SM was constructed and its performance was evaluated using sera from healthy individuals on a Hitachi 7170 autoanalyzer. Results from this assay were highly correlated with theoretical serum SM concentrations estimated by subtracting phosphatidylcholine (PC) and LPC concentrations from that of total phospholipids determined using previously established methods. There was a good correlation between the results of SM assayed by the proposed method and the existing enzymatic method in sera from healthy individuals. Moreover, the proposed method was superior to the existing method in preventing nonspecific reactions with LPC present in sera. The proposed method does not require any pretreatment, uses 2.5 μl of serum samples, and requires only 10 min on an autoanalyzer. This high-throughput method can measure serum SM with sufficient specificity for clinical purposes and is applicable in routine laboratory practice. PMID:26792376

  10. Cellulase hydrolysis of unsorted MSW

    Jensen, Jacob Wagner; Felby, Claus; Jørgensen, Henning

    A recent development in waste management and engineering has shown that the cellulase can be used for the liquefaction of organic fractions in household waste. The focus of this study was to optimize the enzyme hydrolysis of thermally treated municipal solid waste (MSW) by the addition of...

  11. Enzymatic hydrolysis of potato pulp

    Mariusz Lesiecki


    Full Text Available Background. Potato pulp constitutes a complicated system of four types of polysaccharides: cellulose, hemicellulose, pectin and starch. Its composition makes it a potential and attractive raw material for the production of the second generation bioethanol. The aim of this research project was to assess the usefulness of commercial enzymatic preparations for the hydrolysis of potato pulp and to evaluate the effectiveness of hydrolysates obtained in this way as raw materials for ethanol fermentation. Material  and methods. Sterilised potato pulp was subjected to hydrolysis with commercial enzymatic preparations. The effectiveness of the preparations declared as active towards only one fraction of potato pulp (separate amylase, pectinase and cellulase activity and mixtures of these preparations was analysed. The monomers content in hydrolysates was determined using HPLC method. Results.  The application of amylolytic enzymes for potato pulp hydrolysis resulted in the release of only 18% of raw material with glucose as the dominant (77% constituent of the formed product. In addition, 16% galactose was also determined in it. The hydrolysis of the cellulose fraction yielded up to 35% raw material and the main constituents of the obtained hydrolysate were glucose (46% and arabinose (40%. Simultaneous application of amylolytic, cellulolytic and pectinolytic enzymes turned out to be the most effective way of carrying out the process as its efficiency in this case reached 90%. The obtained hydrolysate contained 63% glucose, 25% arabinose and 12% other simple substances. Conclusion. The application of commercial enzymatic preparations made it possible to perform potato pulp hydrolysis with 90% effectiveness. This was achieved by the application of a complex of amylolytic, cellulolytic and pectinolytic enzymes and the hydrolysate obtained in this way contained, primarily, glucose making it a viable substrate for ethanol fermentation.

  12. Sphingomyelin functions as a novel receptor for Helicobacter pylori VacA.

    Vijay R Gupta


    Full Text Available The vacuolating cytotoxin (VacA of the gastric pathogen Helicobacter pylori binds and enters epithelial cells, ultimately resulting in cellular vacuolation. Several host factors have been reported to be important for VacA function, but none of these have been demonstrated to be essential for toxin binding to the plasma membrane. Thus, the identity of cell surface receptors critical for both toxin binding and function has remained elusive. Here, we identify VacA as the first bacterial virulence factor that exploits the important plasma membrane sphingolipid, sphingomyelin (SM, as a cellular receptor. Depletion of plasma membrane SM with sphingomyelinase inhibited VacA-mediated vacuolation and significantly reduced the sensitivity of HeLa cells, as well as several other cell lines, to VacA. Further analysis revealed that SM is critical for VacA interactions with the plasma membrane. Restoring plasma membrane SM in cells previously depleted of SM was sufficient to rescue both toxin vacuolation activity and plasma membrane binding. VacA association with detergent-resistant membranes was inhibited in cells pretreated with SMase C, indicating the importance of SM for VacA association with lipid raft microdomains. Finally, VacA bound to SM in an in vitro ELISA assay in a manner competitively inhibited by lysenin, a known SM-binding protein. Our results suggest a model where VacA may exploit the capacity of SM to preferentially partition into lipid rafts in order to access the raft-associated cellular machinery previously shown to be required for toxin entry into host cells.

  13. Toxin-induced pore formation is hindered by intermolecular hydrogen bonding in sphingomyelin bilayers.

    García-Linares, Sara; Palacios-Ortega, Juan; Yasuda, Tomokazu; Åstrand, Mia; Gavilanes, José G; Martínez-Del-Pozo, Álvaro; Slotte, J Peter


    Sticholysin I and II (StnI and StnII) are pore-forming toxins that use sphingomyelin (SM) for membrane binding. We examined how hydrogen bonding among membrane SMs affected the StnI- and StnII-induced pore formation process, resulting in bilayer permeabilization. We compared toxin-induced permeabilization in bilayers containing either SM or dihydro-SM (lacking the trans Δ(4) double bond of the long-chain base), since their hydrogen-bonding properties are known to differ greatly. We observed that whereas both StnI and StnII formed pores in unilamellar vesicles containing palmitoyl-SM or oleoyl-SM, the toxins failed to similarly form pores in vesicles prepared from dihydro-PSM or dihydro-OSM. In supported bilayers containing OSM, StnII bound efficiently, as determined by surface plasmon resonance. However, StnII binding to supported bilayers prepared from dihydro-OSM was very low under similar experimental conditions. The association of the positively charged StnII (at pH7.0) with unilamellar vesicles prepared from OSM led to a concentration-dependent increase in vesicle charge, as determined from zeta-potential measurements. With dihydro-OSM vesicles, a similar response was not observed. Benzyl alcohol, which is a small hydrogen-bonding compound with affinity to lipid bilayer interfaces, strongly facilitated StnII-induced pore formation in dihydro-OSM bilayers, suggesting that hydrogen bonding in the interfacial region originally prevented StnII from membrane binding and pore formation. We conclude that interfacial hydrogen bonding was able to affect the membrane association of StnI- and StnII, and hence their pore forming capacity. Our results suggest that other types of protein interactions in bilayers may also be affected by hydrogen-bonding origination from SMs. PMID:26975250

  14. N-palmitoyl sphingomyelin bilayers: structure and interactions with cholesterol and dipalmitoylphosphatidylcholine.

    Maulik, P R; Shipley, G G


    The structure and thermotropic properties of N-palmitoyl sphingomyelin (C16:0-SM) and its interaction with cholesterol and dipalmitoylphosphatidylcholine (DPPC) have been studied by differential scanning calorimetry (DSC) and X-ray diffraction methods. DSC of hydrated multi-bilayers of C16:0-SM shows reversible chain-melting transitions. On heating, anhydrous C16:0-SM exhibits an endothermic transition at 75 degrees C (delta H = 4.0 kcal/mol). Increasing hydration progressively lowers the transition temperature (TM) and increases the transition enthalpy (delta H), until limiting values (TM = 41 degrees C, delta H = 7.5 kcal/mol) are observed for hydration values > 25 wt % H2O. X-ray diffraction at temperatures below (29 degrees C) TM show a bilayer gel structure (d = 73.5 A, sharp 4.2 A reflection) for C16:0-SM at full hydration; above TM, at 55 degrees C, a bilayer liquid-crystal phase is present (d = 66.6 A, diffuse 4.6 A reflection). Addition of cholesterol to C16:0-SM bilayers results in a progressive decrease in the enthalpy of the transition at 41 degrees C, and no cooperative transition is detected at > 50 mol % cholesterol. X-ray diffraction shows no difference in the bilayer periodicity, position/width of the wide-angle reflections, or electron density profiles at 29 and 55 degrees C when 50 mol % cholesterol is present. Thus, cholesterol inserts into C16:0-SM bilayers progressively removing the chain-melting transition and changing the structural characteristics of the bilayer. DSC and X-ray diffraction data show that DPPC is completely miscible with C16:0-SM bilayers in both the gel and liquid-crystalline phases; however, 30 mol % C16:0-SM removes the pre-transition exhibited by DPPC. PMID:8672507

  15. Enzymatic hydrolysis of potato pulp

    Mariusz Lesiecki; Wojciech Białas; Grażyna Lewandowicz


    Background. Potato pulp constitutes a complicated system of four types of polysaccharides: cellulose, hemicellulose, pectin and starch. Its composition makes it a potential and attractive raw material for the production of the second generation bioethanol. The aim of this research project was to assess the usefulness of commercial enzymatic preparations for the hydrolysis of potato pulp and to evaluate the effectiveness of hydrolysates obtained in this way as raw materials for ethanol ferment...

  16. Mass Spectrometry Imaging Reveals Elevated Glomerular ATP/AMP in Diabetes/obesity and Identifies Sphingomyelin as a Possible Mediator

    Satoshi Miyamoto


    Full Text Available AMP-activated protein kinase (AMPK is suppressed in diabetes and may be due to a high ATP/AMP ratio, however the quantitation of nucleotides in vivo has been extremely difficult. Via matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI to localize renal nucleotides we found that the diabetic kidney had a significant increase in glomerular ATP/AMP ratio. Untargeted MALDI-MSI analysis revealed that a specific sphingomyelin species (SM(d18:1/16:0 accumulated in the glomeruli of diabetic and high-fat diet-fed mice compared with wild-type controls. In vitro studies in mesangial cells revealed that exogenous addition of SM(d18:1/16:0 significantly elevated ATP via increased glucose consumption and lactate production with a consequent reduction of AMPK and PGC1α. Furthermore, inhibition of sphingomyelin synthases reversed these effects. Our findings suggest that AMPK is reduced in the diabetic kidney due to an increase in the ATP/AMP ratio and that SM(d18:1/16:0 could be responsible for the enhanced ATP production via activation of the glycolytic pathway.


    Previous studies show the enrichment of mammalian brain with neutral sphingomyelin specific phospholipase C (ceramide-phosphocholine phosphodiesterase, EC; N-Sase). The objective of this study was to evaluate the subcellular N-Sase activity in striatum, hippocampus, and frontal cortex. Resu...

  18. Techno-economical evaluation of lignocellulose hydrolysis

    Mirsch, Mikaela


    The economic dependency on fossil fuels affects the climate and environment, which drives the fuel research on the largest known renewable carbohydrate source: fermentable sugars from lignocellulose. Several fermentable sugars exist in lignicellulosic materials, but are not accessible for efficient use without pretreatment and hydrolysis. Enzymatic hydrolysis is typically used. Enzymatic hydrolysis has a high selectivity and is performed in mild conditions, but the cost of...

  19. Enzymatic hydrolysis of plant extracts containing inulin

    Guiraud, J.P.; Galzy, P.


    Inulin-rich extracts of chicory and Jerusalem artichoke are a good potential source of fructose. Total enzymatic hydrolysis of these extracts can be effected by yeast inulinases (EC Chemical prehydrolysis is unfavourable. Enzymatic hydrolysis has advantages over chemical hydrolysis: it does not produce a dark-coloured fraction or secondary substances. It is possible to envisage the preparation of high fructose syrups using this process. (Refs. 42).

  20. Kinetics of enzymatic hydrolysis of methyl ricinoleate

    Neeharika, T. S.V.R.; Lokesh, P.; Prasanna Rani, K. N.; Prathap Kumar, T.; Prasad, R. B.N.


    Ricinoleic acid is an unsaturated hydroxy fatty acid that naturally occurs in castor oil in proportions of up to 85–90%. Ricinoleic acid is a potential raw material and finds several applications in coatings, lubricant formulations and pharmaceutical areas. Enzymatic hydrolysis of castor oil is preferred over conventional hydrolysis for the preparation of ricinoleic acid to avoid estolide formation. A kinetics analysis of the enzymatic hydrolysis of Methyl Ricinoleate in the presence of Candi...

  1. X-ray diffraction and calorimetric study of N-lignoceryl sphingomyelin membranes.

    Maulik, P R; Shipley, G G


    Differential scanning calorimetry and x-ray diffraction have been used to investigate hydrated multibilayers of N-lignoceryl sphingomyelin (C24:0-SM) in the hydration range 0-75 wt % H2O. Anhydrous C24:0-SM exhibits a single endothermic transition at 81.3 degrees C (delta H = 3.6 kcal/mol). At low hydration (12.1 wt % H2O), three different endothermic transitions are observed: low-temperature transition (T1) at 39.4 degrees C (transition enthalpy (delta H1) = 2.8 kcal/mol), intermediate-temperature transition (T2) at 45.5 degrees C, and high-temperature transition (T3) at 51.3 degrees C (combined transition enthalpy (delta H2 + 3) = 5.03 kcal/mol). On increasing hydration, all three transition temperatures of C24:0-SM decrease slightly to reach limiting values of 36.7 degrees C (T1), 44.4 degrees C (T2), and 48.4 degrees C (T3) at approximately 20 wt % H2O. At 22 degrees C (below T1), x-ray diffraction of C24:0-SM at different hydration levels shows two wide-angle reflections, a sharp one at 1/4.2 A-1 and a more diffuse one at 1/4.0 A-1 together with lamellar reflections corresponding to bilayer periodicities increasing from d = 65.4 A to a limiting value of 71.1 A. Electron density profiles show a constant bilayer thickness dp-p approximately 50 A. In contrast, at 40 degrees C (between T1 and T2) a single sharp wide-angle reflection at approximately 1/4.2 A-1 is observed. The lamellar reflections correspond to a larger bilayer periodicity (increasing from d = 69.3-80.2 A) and there is some increase in dp-p (52-56 A) with hydration. These structural parameters,together with calculated lipid thickness and molecular area considerations, suggest that the low temperature endotherm(T1) of hydrated C24:0-SM corresponds to a transition from a tilted, gel state (Gel I) with partially interdigitated chains to an untilted, or less tilted, gel state (Gel 11). At 600C (above T3), the usual liquid-crystalline La bilayer structure (d = 59.5-66.3A; dp p -46 A) is present at all

  2. Rate of Hydrolysis of Tertiary Halogeno Alkanes

    Pritchard, D. R.


    Describes an experiment to measure the relative rate of hydrolysis of the 2-x-2 methylpropanes, where x is bromo, chloro or iodo. The results are plotted on a graph from which the relative rate of hydrolysis can be deduced. (Author/GA)

  3. Hydrolysis of hafnium nitrides and carbides

    Hydrolysis of Hafnium Nitrides and Carbides. The hydrolytic behavior of Hafnium mononitride and monocarbide has been studied and compared with that of Titanium and Zirconium nitrides and carbides. In the case of hydrolysis of HfN the gaseous products were H2, N2 and a small amount of NO, and the liquid product was NH3, as in the case of TiN and ZrN. In isothermal hydrolysis the principal product was NH3 at temperatures lower than 8000C, which was replaced by N2 at temperatures higher than 9000C. In this respect HfN was similar to ZrN, but not to TiN which produced mainly N2 even by hydrolysis at 8000C. The products of hydrolysis of HfC were found to be CO, CO2, H2 and a small amount of CH4 also as in the case of TiC and ZrC. In the isothermal hydrolysis of HfC it was observed that a large amount of H2 evolved at the early stage of the hydrolysis while CO2 continued to evolve with some amount of H2 even after the ceasing of CO evolution. From analysis of the hydrolytic behavior the solid residue after the hydrolysis of HfC was considered to contain some waxes (Csub(n)Hsub(m)). It was suggested that the carbide of the element of smaller atomic number (Ti) would tend to form oxygen compounds (CO, CO2) while the carbide of the element of larger atomic number (Zr, Hf) hydrogen compounds(Csub(n)Hsub(m)), since ThC and UC formed only hydrocarbons and H2 by hydrolysis. This suggestion was also valid to nitride. (auth.)

  4. Chlamydia trachomatis interrupts an exocytic pathway to acquire endogenously synthesized sphingomyelin in transit from the Golgi apparatus to the plasma membrane.

    Hackstadt, T; Rockey, D D; Heinzen, R A; Scidmore, M. A.


    Chlamydia trachomatis acquires C6-NBD-sphingomyelin endogenously synthesized from C6-NBD-ceramide and transported to the vesicle (inclusion) in which they multiply. Here we explore the mechanisms of this unusual trafficking and further characterize the association of the chlamydial inclusion with the Golgi apparatus. Endocytosed chlamydiae are trafficked to the Golgi region and begin to acquire sphingolipids from the host within a few hours following infection. The transport of NBD-sphingolip...

  5. Thioglycoside hydrolysis catalyzed by β-glucosidase

    Sweet almond β-glucosidase (EC has been shown to have significant thioglycohydrolase activity. While the Km values for the S- and O-glycosides are similar, the kcat values are about 1000-times lower for the S-glycosides. Remarkably, the pH-profile for kcat/Km for hydrolysis of p-nitrophenyl thioglucoside (pNPSG) shows the identical dependence on a deprotonated carboxylate (pKa 4.5) and a protonated group (pKa 6.7) as does the pH-profile for hydrolysis of the corresponding O-glycoside. Not surprisingly, in spite of the requirement for the presence of this protonated group in catalytically active β-glucosidase, thioglucoside hydrolysis does not involve general acid catalysis. There is no solvent kinetic isotope effect on the enzyme-catalyzed hydrolysis of pNPSG

  6. Hemicellulose hydrolysis catalysed by solid acids

    P.D.. Carà; M. Pagliaro; A. Elmekawy; D.R. Brown; P. Verschuren; N.R. Shiju; G. Rothenberg


    Depolymerising hemicellulose into platform sugar molecules is a key step in developing the concept of an integrated biorefinery. This reaction is traditionally catalysed by either enzymes or homogeneous mineral acids. We compared various solid catalysts for hemicellulose hydrolysis, running reaction

  7. Hydrolysis of isocyanic acid on SCR catalysts

    Elsener, M.; Kleemann, M.; Koebel, M. [Paul Scherrer Inst. (PSI), Villigen (Switzerland)


    Standard SCR catalysts possess high activity for the hydrolysis of HNCO and thus explain the suitability of urea as a selective reducing agent for NO{sub x}. At high space velocities HNCO-slip can get perceptible over the entire temperature range. This can be attributed to the fact that the temperature dependence is strong for the SCR reaction, but weak for the hydrolysis reaction. (author) 3 figs., 5 refs.


    M. F. CUSTÓDIO; A. J. GOULART; D. P. MARQUES; R.C. Giordano; R. L. C. Giordano; R. MONTI


    This work presents a method for adding value to cheese whey residues by whey proteins hydrolysis, using trypsin, chymotrypsin and carboxypeptidase A as catalysts. Sweet cheese whey was dialyzed and filtered in kaolin. Lactose and protein contents were analyzed after each step. The activities of bovine pancreas trypsin and chymotrypsin were measured at different pHs and temperatures. The optimal pH for the hydrolysis of whey proteins was 9.0 for both enzymes. Optima te...

  9. Enzymatic hydrolysis of pretreated soybean straw

    In order to produce lactic acid, from agricultural residues such as soybean straw, which is a raw material for biodegradable plastic production, it is necessary to decompose the soybean straw into soluble sugars. Enzymatic hydrolysis is one of the methods in common use, while pretreatment is the effective way to increase the hydrolysis rate. The optimal conditions of pretreatment using ammonia and enzymatic hydrolysis of soybean straw were determined. Compared with the untreated straw, cellulose in straw pretreated by ammonia liquor (10%) soaking for 24 h at room temperature increased 70.27%, whereas hemicellulose and lignin in pretreated straw decreased to 41.45% and 30.16%, respectively. The results of infrared spectra (IR), scanning electron microscope (SEM) and X-ray diffraction (XRD) analysis also showed that the structure and the surface of the straw were changed through pretreatment that is in favor of the following enzymatic hydrolysis. maximum enzymatic hydrolysis rate of 51.22% was achieved at a substrate concentration of 5% (w/v) at 50 deg. C and pH 4.8 using cellulase (50 fpu/g of substrate) for 36 h


    Keikhosro Karimi


    Full Text Available This article reviews developments in the technology for ethanol produc-tion from lignocellulosic materials by “enzymatic” processes. Several methods of pretreatment of lignocelluloses are discussed, where the crystalline structure of lignocelluloses is opened up, making them more accessible to the cellulase enzymes. The characteristics of these enzymes and important factors in enzymatic hydrolysis of the cellulose and hemicellulose to cellobiose, glucose, and other sugars are discussed. Different strategies are then described for enzymatic hydrolysis and fermentation, including separate enzymatic hydrolysis and fermentation (SHF, simultaneous saccharification and fermentation (SSF, non-isothermal simultaneous saccharification and fermentation (NSSF, simultaneous saccharification and co-fermentation (SSCF, and consolidated bioprocessing (CBP. Furthermore, the by-products in ethanol from lignocellulosic materials, wastewater treatment, commercial status, and energy production and integration are reviewed.




    Full Text Available The yield, productivity and cost for the enzymatic hydrolysis of cellulose to glucoseare crucial for the production of second generation ethanol. In the first study wehave evaluated the activity of several commercial cellulolytic enzymes and a crudeextract of a local strain of Trichoderma viride. The load used was 15 U ofcellulase/gram cellulose and 90 U of cellobiase/gram cellulose. The hydrolysis wascarried out at 50oC and pH 4,8 for 96 hours. The best cellulose hydrolysis yield of58% was obtained with the cocktail formed of crude cellulases from T. virideCMIT3.5 combined with Novozyme 188. This cocktail was used in the second study,when alkaline-steam pretreated wheat straw and corn stover where hydrolyzed at pH4,8 for 96 hours. The temperature was set at 50oC and 40oC. The hydrolysis at lowertemperature was tested for a future experiment of simultaneous hydrolysis andfermentation. An enzymatic assay using glucose-6-phosphate dehydrogenase wasused to determine exclusively glucose, instead of wide-range sugar DNS assay.Reporting to 100 grams of wet pretreated biomass, the following results wereobtained: 14.4 g% glucose for corn stover at 50oC and 13,0 g% at 40oC; 13,1 g%glucose for wheat straw at 50oC and 10.3 g% at 40oC. Considering that wheat strawcontain 36.6% glucose-based carbohydrates, the hydrolysis yields are between39.3% and 28.1%. Further studies, concerning the optimal parameters for cellulasecocktail will be made.

  12. Enzymatic Hydrolysis Conditions for Egg White Proteins

    Chi, Yujie; Tian, Bo; Sun, Bo; Guo, Mingruo


    The enzymatic hydrolysis of proteins in egg white by Alcalase was systematically studied through dual quadratic rotary, orthogonal and regressive design. The optimum conditions of hydrolysis were determined. The results showed that the optimum temperature was 68.5℃, pH 8.21 at the substrate concentration of 5.5%. The regression equation, Y=42.6994+0.3344X1+7.53X2-0.0086X1X2-0.001X21-0.4726X22 (Y-nitrogen recovery rate, NR; X1-enzyme concentration /substrate concentration, E/S; X2-hydrolytic t...

  13. Clearance of Hepatic Sphingomyelin by Olipudase Alfa Is Associated With Improvement in Lipid Profiles in Acid Sphingomyelinase Deficiency.

    Thurberg, Beth L; Wasserstein, Melissa P; Jones, Simon A; Schiano, Thomas D; Cox, Gerald F; Puga, Ana Cristina


    Acid sphingomyelinase deficiency (ASMD; Niemann-Pick disease type A and B) is a lysosomal storage disorder characterized by abnormal intracellular sphingomyelin (SM) accumulation. Prominent liver involvement results in hepatomegaly, fibrosis/cirrhosis, abnormal liver chemistries, and a proatherogenic lipid profile. Olipudase alfa (recombinant human ASM) is in clinical development as an investigational enzyme replacement therapy for the non-neurological manifestations of ASMD. In a phase 1b study conducted to evaluate the safety and tolerability of within-patient dose escalation with olipudase alfa, measurement of SM levels in liver biopsies was used as a pharmacodynamic biomarker of substrate burden. Five adult patients with non neuronopathic ASMD received escalating doses of olipudase alfa every 2 weeks for 26 weeks. Liver biopsies obtained at baseline and 26 weeks after treatment were evaluated for SM storage by histomorphometric analysis, biochemistry, and electron microscopy. Biopsies were also assessed for inflammation and fibrosis, and for the association of SM levels with liver volume, liver function tests, and lipid profiles. At baseline, SM storage present in Kupffer cells and hepatocytes ranged from 9.8% to 53.8% of the microscopic field. After 26 weeks of treatment, statistically significant reductions in SM (P<0.0001) measured by morphometry were seen in 4 patients with evaluable liver biopsies. The 26-week biopsy of the fifth patient was insufficient for morphometric quantitation. Posttreatment SM levels ranged from 1.2% to 9.5% of the microscopic field, corresponding to an 84% to 92% relative reduction from baseline. Improvements in liver volume, liver function tests, and lipid profiles were also observed. This study illustrates the utility of SM assessment by liver biopsy as a pharmacodynamic biomarker of disease burden in these patients. PMID:27340749

  14. Sphingomyelin phosphodiesterase-1 (SMPD1 coding variants do not contribute to low levels of high-density lipoprotein cholesterol

    Genest Jacques


    Full Text Available Abstract Background Niemann-Pick disease type A and B is caused by a deficiency of acid sphingomyelinase due to mutations in the sphingomyelin phosphodiesterase-1 (SMPD1 gene. In Niemann-Pick patients, SMPD1 gene defects are reported to be associated with a severe reduction in plasma high-density lipoprotein (HDL cholesterol. Methods Two common coding polymorphisms in the SMPD1 gene, the G1522A (G508R and a hexanucleotide repeat sequence within the signal peptide region, were investigated in 118 unrelated subjects of French Canadian descent with low plasma levels of HDL-cholesterol (th percentile for age and gender-matched subjects. Control subjects (n = 230 had an HDL-cholesterol level > the 25th percentile. Results For G1522A the frequency of the G and A alleles were 75.2% and 24.8% respectively in controls, compared to 78.6% and 21.4% in subjects with low HDL-cholesterol (p = 0.317. The frequency of 6 and 7 hexanucleotide repeats was 46.2% and 46.6% respectively in controls, compared to 45.6% and 49.1% in subjects with low HDL-cholesterol (p = 0.619. Ten different haplotypes were observed in cases and controls. Overall haplotype frequencies in cases and controls were not significantly different. Conclusion These results suggest that the two common coding variants at the SMPD1 gene locus are not associated with low HDL-cholesterol levels in the French Canadian population.

  15. Sustained Epigenetic Drug Delivery Depletes Cholesterol-Sphingomyelin Rafts from Resistant Breast Cancer Cells, Influencing Biophysical Characteristics of Membrane Lipids.

    Raghavan, Vijay; Vijayaraghavalu, Sivakumar; Peetla, Chiranjeevi; Yamada, Masayoshi; Morisada, Megan; Labhasetwar, Vinod


    Cell-membrane lipid composition can greatly influence biophysical properties of cell membranes, affecting various cellular functions. We previously showed that lipid synthesis becomes altered in the membranes of resistant breast cancer cells (MCF-7/ADR); they form a more rigid, hydrophobic lipid monolayer than do sensitive cell membranes (MCF-7). These changes in membrane lipids of resistant cells, attributed to epigenetic aberration, significantly affected drug transport and endocytic function, thus impacting the efficacy of anticancer drugs. The present study's objective was to determine the effects of the epigenetic drug, 5-aza-2'-deoxycytidine (DAC), delivered in sustained-release nanogels (DAC-NGs), on the composition and biophysical properties of membrane lipids of resistant cells. Resistant and sensitive cells were treated with DAC in solution (DAC-sol) or DAC-NGs, and cell-membrane lipids were isolated and analyzed for lipid composition and biophysical properties. In resistant cells, we found increased formation of cholesterol-sphingomyelin (CHOL-SM) rafts with culturing time, whereas DAC treatment reduced their formation. In general, the effect of DAC-NGs was greater in changing the lipid composition than with DAC-sol. DAC treatment also caused a rise in levels of certain phospholipids and neutral lipids known to increase membrane fluidity, while reducing the levels of certain lipids known to increase membrane rigidity. Isotherm data showed increased lipid membrane fluidity following DAC treatment, attributed to decrease levels of CHOL-SM rafts (lamellar beta [Lβ] structures or ordered gel) and a corresponding increase in lipids that form lamellar alpha-structures (Lα, liquid crystalline phase). Sensitive cells showed marginal or insignificant changes in lipid profile following DAC-treatment, suggesting that epigenetic changes affecting lipid biosynthesis are more specific to resistant cells. Since membrane fluidity plays a major role in drug transport

  16. A raft-associated species of phosphatidylethanolamine interacts with cholesterol comparably to sphingomyelin. A Langmuir-Blodgett monolayer study.

    Michal Grzybek

    Full Text Available BACKGROUND: Specific interactions between sphingomyelin (SM and cholesterol (Ch are commonly believed to play a key role in the formation of rafts in the biological membranes. A weakness of this model is the implication that these microdomains are confined to the outer bilayer leaflet. The cytoplasmic leaflet, which contains the bulk of phosphatidylethanolamine (PE, phosphatidylserine (PS and phosphatidylinositol (PI, is thought also to harbour half of the membrane cholesterol. Moreover, SLPE (1-stearoyl-2-linoleoyl-sn-glycero-3-phosphatidyl-ethanolamine has recently been shown to be enriched in isolated detergent-resistant membranes (DRM, and this enrichment was independent of the method of isolation of DRM. METHODOLOGY/PRINCIPAL FINDINGS: Here we present quantitative evidence coming from Langmuir-Blodgett monolayer experiments that SLPE forms complex with Ch similar to that between SM and Ch. The energies of these interactions as calculated form the monolayer studies are highly negative. FRAP analysis showed that NBD-Ch recovery was similar in liposomes composed of DOPC/Ch SM or SLPE but not DPPE, providing further evidence that SLPE may form an l(o phase in the presence of high Ch concentration. Experiments on the solubility of DOPC liposomes containing DPPE/Ch (1ratio1, SM/Ch (1ratio1 or SLPE/Ch (1ratio1 showed the presence of Triton X-100 insoluble floating fraction (TIFF in the case of SM/Ch or SLPE/Ch but not in DPPE/Ch containing liposomes. Quantitative determination of particular lipid species in the TIFF fraction confirms the conclusion that SLPE (or similar PE species could be an important constituent of the inner leaflet raft. CONCLUSION: Such interactions suggest a possible existence of inner-leaflet nanoscale assemblies composed of cholesterol complexes with SLPE or similar unsaturated PE species.

  17. Characterization of the role of sphingomyelin synthase 2 in glucose metabolism in whole-body and peripheral tissues in mice.

    Sugimoto, Masayuki; Shimizu, Yoichi; Zhao, Songji; Ukon, Naoyuki; Nishijima, Ken-Ichi; Wakabayashi, Masato; Yoshioka, Takeshi; Higashino, Kenichi; Numata, Yoshito; Okuda, Tomohiko; Tamaki, Nagara; Hanamatsu, Hisatoshi; Igarashi, Yasuyuki; Kuge, Yuji


    Sphingomyelin synthase 2 (SMS2) is a proposed potential therapeutic target for obesity and insulin resistance. However, the contributions of SMS2 to glucose metabolism in tissues and its possible therapeutic mechanisms remain unclear. Thus, to determine whole-body glucose utilization and the contributions of each insulin-targeted tissue to glucose uptake, we performed a glucose kinetics study, using the radiolabeled glucose analog (18)F-2-fluoro-2-deoxy-D-glucose ((18)F-FDG), in wild-type (WT) and SMS2 knockout (KO) mice. Insulin signaling was enhanced in the liver, white adipose tissue and skeletal muscle of SMS2 KO mice compared with those of WT mice. In addition, compared with in WT mice, blood clearance of (18)F-FDG was accelerated in SMS2 KO mice when they were fed either a normal or a high fat diet. (18)F-FDG uptake was also increased in insulin-targeted tissues such as skeletal muscle in the SMS2 KO mice. Whereas skeletal muscle sphingolipid content was not clearly affected, plasma levels of very long-chain fatty acid (VLCFA)-containing ceramides were markedly increased in SMS2 KO mice, compared with in WT mice. We also generated liver-conditional SMS2 KO mice and performed glucose and insulin tolerance tests on mice with a high fat diet. However, no significant effect was observed. Thus, our study provided evidence that genetic inhibition of SMS2 elevated glucose clearance through activation of glucose uptake into insulin-targeted tissues such as skeletal muscle by a mechanism independent of hepatic SMS2. Our findings further indicate that this occurs, at least in part, via indirect mechanisms such as elevation of VLCFA-containing ceramides. PMID:27151272




    Full Text Available

    This work presents a method for adding value to cheese whey residues by whey proteins hydrolysis, using trypsin, chymotrypsin and carboxypeptidase A as catalysts. Sweet cheese whey was dialyzed and filtered in kaolin. Lactose and protein contents were analyzed after each step. The activities of bovine pancreas trypsin and chymotrypsin were measured at different pHs and temperatures. The optimal pH for the hydrolysis of whey proteins was 9.0 for both enzymes. Optima temperatures were 60ºC for trypsin, and 50ºC for chymotrypsin. Trypsin exhibited typical Michaelis-Menten behavior, but chymotrypsin did not. Electrophoretic analysis showed that neither trypsin nor chymotrypsin alone hydrolyzed whey proteins in less than three hours. Hydrolysis rates of -lactalbumin by trypsin, and of bovine serum albumin by chymotrypsin were low. When these enzymes were combined, however, all protein fractions were attacked and rates of hydrolysis were enhanced by one order of magnitude. The addition of carboxypeptidase A to the others enzymes did not improve the process yield.

  19. Monitoring enzymatic ATP hydrolysis by EPR spectroscopy

    Hacker, Stephan M.; Hintze, Christian; Marx, Andreas; Drescher, Malte


    An adenosine triphosphate (ATP) analogue modified with two nitroxide radicals is developed and employed to study its enzymatic hydrolysis by electron paramagnetic resonance spectroscopy. For this application, we demonstrate that EPR holds the potential to complement fluorogenic substrate analogues in monitoring enzymatic activity.

  20. Mechanisms of lactone hydrolysis in acidic conditions.

    Gómez-Bombarelli, Rafael; Calle, Emilio; Casado, Julio


    The acid-catalyzed hydrolysis of linear esters and lactones was studied using a hybrid supermolecule-polarizable continuum model (PCM) approach including up to six water molecules. The compounds studied included two linear esters, four β-lactones, two γ-lactones, and one δ-lactone: ethyl acetate, methyl formate, β-propiolactone, β-butyrolactone, β-isovalerolactone, diketene (4-methyleneoxetan-2-one), γ-butyrolactone, 2(5H)-furanone, and δ-valerolactone. The theoretical results are in good quantitative agreement with the experimental measurements reported in the literature and also in excellent qualitative agreement with long-held views regarding the nature of the hydrolysis mechanisms at molecular level. The present results help to understand the balance between the unimolecular (A(AC)1) and bimolecular (A(AC)2) reaction pathways. In contrast to the experimental setting, where one of the two branches is often occluded by the requirement of rather extreme experimental conditions, we have been able to estimate both contributions for all the compounds studied and found that a transition from A(AC)2 to A(AC)1 hydrolysis takes place as acidity increases. A parallel work addresses the neutral and base-catalyzed hydrolysis of lactones. PMID:23731203

  1. Microwave-assisted Weak Acid Hydrolysis of Proteins

    Miyeong Seo


    Full Text Available Myoglobin was hydrolyzed by microwave-assisted weak acid hydrolysis with 2% formic acid at 37 oC, 50 oC, and100 oC for 1 h. The most effective hydrolysis was observed at 100 oC. Hydrolysis products were investigated using matrixassistedlaser desorption/ionization time-of-flight mass spectrometry. Most cleavages predominantly occurred at the C-termini ofaspartyl residues. For comparison, weak acid hydrolysis was also performed in boiling water for 20, 40, 60, and 120 min. A 60-min weak acid hydrolysis in boiling water yielded similar results as a 60-min microwave-assisted weak acid hydrolysis at100 oC. These results strongly suggest that microwave irradiation has no notable enhancement effect on acid hydrolysis of proteinsand that temperature is the major factor that determines the effectiveness of weak acid hydrolysis.

  2. Corn Gluten Hydrolysis By Alcalase: Effects of Process Parameters on Hydrolysis, Solubilization and Enzyme Inactivation

    Kilic-Apar, D.; Ozbek, B.


    The aim of this study was to investigate the influences of substrate concentration, enzyme concentration, temperature and pH on hydrolysis and solubilization of corn gluten as well as enzyme stability. The corn gluten was hydrolyzed by Alcalase enzyme (a bacterial protease produced by a selected strain of Bacillus Licheniformis) that was chosen among five commercial enzymes examined. The optimum process conditions for hydrolysis and solubilization were obtained as 30 g L-1 substrate mass conc...

  3. Modeling and analysis of the enzymatic hydrolysis of lignocellulosic substrates

    Sola Saura, Alaia


    Simultaneous saccharification and fermentation (SSF) and simultaneous saccharification and cofermentation (SSCF) are two process options for production of ethanol from lignocellulosic substrates that are superior to separate hydrolysis and fermentation (SHF). The principal benefits of performing the enzymatic hydrolysis together with the fermentation, instead of in a separate step after the hydrolysis as SHF does, are the reduced end-product inhibition of the enzymatic hydrolysis, and t...

  4. Regulation of SREBPs by Sphingomyelin in Adipocytes via a Caveolin and Ras-ERK-MAPK-CREB Signaling Pathway.

    Nehman Makdissy

    Full Text Available Sterol response element binding protein (SREBP is a key transcription factor in insulin and glucose metabolism. We previously demonstrated that elevated levels of membrane sphingomyelin (SM were related to peroxisome proliferator-activated receptor-γ (PPARγ, which is a known target gene of SREBP-1 in adipocytes. However, the role of SM in SREBP expression in adipocytes remains unknown. In human abdominal adipose tissue from obese women with various concentrations of fasting plasma insulin, SREBP-1 proteins decreased in parallel with increases in membrane SM levels. An inverse correlation was found between the membrane SM content and the levels of SREBP-1c/ERK/Ras/PPARγ/CREB proteins. For the first time, we demonstrate the effects of SM and its signaling pathway in 3T3-F442A adipocytes. These cells were enriched or unenriched with SM in a range of concentrations similar to those observed in obese subjects by adding exogenous natural SMs (having different acyl chain lengths or by inhibiting neutral sphingomyelinase. SM accumulated in caveolae of the plasma membrane within 24 h and then in the intracellular space. SM enrichment decreased SREBP-1 through the inhibition of extracellular signal-regulated protein kinase (ERK but not JNK or p38 mitogen-activated protein kinase (MAPK. Ras/Raf-1/MEK1/2 and KSR proteins, which are upstream mediators of ERK, were down-regulated, whereas SREBP-2/caveolin and cholesterol were up-regulated. In SM-unmodulated adipocytes treated with DL-1-Phenyl-2-Palmitoylamino-3-morpholino-1-propanol (PPMP, where the ceramide level increased, the expression levels of SREBPs and ERK were modulated in an opposite direction relative to the SM-enriched cells. SM inhibited the insulin-induced expression of SREBP-1. Rosiglitazone, which is an anti-diabetic agent and potent activator of PPARγ, reversed the effects of SM on SREBP-1, PPARγ and CREB. Taken together, these findings provide novel insights indicating that excess

  5. Chlamydia trachomatis co-opts GBF1 and CERT to acquire host sphingomyelin for distinct roles during intracellular development.

    Cherilyn A Elwell


    Full Text Available The strain designated Chlamydia trachomatis serovar that was used for experiments in this paper is Chlamydia muridarum, a species closely related to C. trachomatis (and formerly termed the Mouse Pneumonitis strain of C. trachomatis. [corrected]. The obligate intracellular pathogen Chlamydia trachomatis replicates within a membrane-bound inclusion that acquires host sphingomyelin (SM, a process that is essential for replication as well as inclusion biogenesis. Previous studies demonstrate that SM is acquired by a Brefeldin A (BFA-sensitive vesicular trafficking pathway, although paradoxically, this pathway is dispensable for bacterial replication. This finding suggests that other lipid transport mechanisms are involved in the acquisition of host SM. In this work, we interrogated the role of specific components of BFA-sensitive and BFA-insensitive lipid trafficking pathways to define their contribution in SM acquisition during infection. We found that C. trachomatis hijacks components of both vesicular and non-vesicular lipid trafficking pathways for SM acquisition but that the SM obtained from these separate pathways is being utilized by the pathogen in different ways. We show that C. trachomatis selectively co-opts only one of the three known BFA targets, GBF1, a regulator of Arf1-dependent vesicular trafficking within the early secretory pathway for vesicle-mediated SM acquisition. The Arf1/GBF1-dependent pathway of SM acquisition is essential for inclusion membrane growth and stability but is not required for bacterial replication. In contrast, we show that C. trachomatis co-opts CERT, a lipid transfer protein that is a key component in non-vesicular ER to trans-Golgi trafficking of ceramide (the precursor for SM, for C. trachomatis replication. We demonstrate that C. trachomatis recruits CERT, its ER binding partner, VAP-A, and SM synthases, SMS1 and SMS2, to the inclusion and propose that these proteins establish an on-site SM biosynthetic

  6. Effect of gelatinization and hydrolysis conditions on the selectivity of starch hydrolysis with alpha-amylase from Bacillus licheniformis

    Baks, T.; Bruins, M.E.; Matser, A.M.; Janssen, A.E.M.; Boom, R.M.


    Enzymatic hydrolysis of starch can be used to obtain various valuable hydrolyzates with different compositions. The effects of starch pretreatment, enzyme addition point, and hydrolysis conditions on the hydrolyzate composition and reaction rate during wheat starch hydrolysis with ¿-amylase from Bac

  7. Enzymatic hydrolysis of corn bran arabinoxylan

    Agger, Jane

    This thesis concerns enzymatic hydrolysis of corn bran arabinoxylan. The work has focused on understanding the composition and structure of corn bran with specific interest in arabinoxylan with the main purpose of targeting enzymatic hydrolysis for increased yields. Corn bran has been used as a...... model substrate because it represents a readily available agroindustrial side product with upgrading potentials. Corn bran originates from the wet-milling process in corn starch processing, is the outmost layers of the corn kernel and is particularly rich in pentose monosaccharides comprising the major...... components of arabinoxylan. Corn bran is one of the most recalcitrant cereal byproducts with arabinoxylans of particular heterogeneous nature. It is also rich in feruloyl derived substitutions, which are responsible for extensive cross-linking between arabinoxylan molecules and thereby participate in a...

  8. Modeling and analysis of calcium bromide hydrolysis

    Lottes, Steven A.; Lyczkowski, Robert W.; Panchal, Chandrakant B.; Doctor, Richard D. [Energy Systems Division, Argonne National Laboratory, 9700 S. Cass Avenue, Argonne, IL 60439 (United States)


    The main focus of this paper is the modeling, simulation, and analysis of the calcium bromide hydrolysis reactor stage in the calcium-bromine thermochemical water-splitting cycle for nuclear hydrogen production. One reactor concept is to use a spray of calcium bromide into steam, in which the heat of fusion supplies the heat of reaction. Droplet models were built up in a series of steps incorporating various physical phenomena, including droplet flow, heat transfer, phase change, and reaction, separately. Given the large heat reservoir contained in a pool of molten calcium bromide that allows bubbles to rise easily, using a bubble column reactor for the hydrolysis appears to be a feasible and promising alternative to the spray reactor concept. The two limiting cases of bubble geometry, spherical and spherical-cap, are considered in the modeling. Results for both droplet and bubble modeling with COMSOL MULTIPHYSICS trademark are presented, with recommendations for the path forward. (author)




    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.




    Full Text Available Hydrolysis of the hemicellulosic fraction of sugarcane bagasse by sulphuric acid was performed in laboratory (25 mL and semi-pilot (25 L reactors under different conditions of temperature, time and acid concentration. On the laboratory scale, the three highest recovery yields were obtained at: 140ºC for 10 min with 100 mgacid/gdm (yield=73.4%; 140ºC for 20 min with 100 mgacid/gdm (yield=73.9% and 150ºC for 20 min with 70 mgacid/gdm (yield=71.8%. These conditions were also used for hydrolysis in a semi-pilot reactor, and the highest xylose recovery yield (83.3% was obtained at 140ºC for 20 min with 100 mgacid/gdm

  11. Enzymatic Hydrolysis of Alkaline Pretreated Coconut Coir

    Akbarningrum Fatmawati


    Full Text Available The purpose of this research is to study the effect of concentration and temperature on the cellulose and lignin content, and the reducing sugars produced in the enzymatic hydrolysis of coconut coir. In this research, the coconut coir is pretreated using 3%, 7%, and 11% NaOH solution at 60oC, 80oC, and 100oC. The pretreated coir were assayed by measuring the amount of cellulose and lignin and then hydrolysed using Celluclast and Novozyme 188 under various temperature (30oC, 40oC, 50oC and pH (3, 4, 5. The hydrolysis results were assayed for the reducing sugar content. The results showed that the alkaline delignification was effective to reduce lignin and to increase the cellulose content of the coir. The best delignification condition was observed at 11% NaOH solution and 100oC which removed 14,53% of lignin and increased the cellulose content up to 50,23%. The best condition of the enzymatic hydrolysis was obtained at 50oC and pH 4 which produced 7,57 gr/L reducing sugar. © 2013 BCREC UNDIP. All rights reservedReceived: 2nd October 2012; Revised: 31st January 2013; Accepted: 6th February 2013[How to Cite: Fatmawati, A., Agustriyanto, R., Liasari, Y. (2013. Enzymatic Hydrolysis of Alkaline Pre-treated Coconut Coir. Bulletin of Chemical Reaction Engineering & Catalysis, 8 (1: 34-39 (doi:10.9767/bcrec.8.1.4048.34-39[Permalink/DOI:] | View in  |

  12. Hemicellulose hydrolysis catalysed by solid acids

    Carà, P.D..; Pagliaro, M.; Elmekawy, A.; Brown, D R; Verschuren, P.; Shiju, N.R.; Rothenberg, G.


    Depolymerising hemicellulose into platform sugar molecules is a key step in developing the concept of an integrated biorefinery. This reaction is traditionally catalysed by either enzymes or homogeneous mineral acids. We compared various solid catalysts for hemicellulose hydrolysis, running reactions in water, under neutral pH and relatively mild temperature and pressure (120 degrees C and 10 bar) conditions. Sulphonated resins are highly active, but they leach out sulphonic groups. Sulphonat...

  13. Enzymatic hydrolysis of PTT polymers and oligomers

    Eberl, A.; Heumann, Sonja; Kotek, R.; Kaufmann, F; Mitscher, S.; Paulo, Artur Cavaco; Gübitz, Georg M.


    Oligomers and polymers (film, fabrics) of the linear aromatic polyester poly(trimethylene terephthalate) (PTT) were treated with polyesterases from Thermomyces lanuginosus, Penicillium citrinum, Thermobifida fusca and Fusarium solani pisi. The cutinase from T. fusca was found to release the highest amounts of hydrolysis products from PTT materials and was able to open and hydrolyse a cyclic PTT dimer according to RP-HPLC–UV detection. In contrast, the lipase from T. lanuginosus also showed ac...

  14. Phytate hydrolysis by germfree and conventional rats.

    Wise, A; Gilburt, D J


    Phytic acid is naturally occurring compound that reduces intestinal absorption of many metals. Early work suggests that some dietary phytate may be hydrolyzed in the large intestines by bacteria, but more recently nutritionists have suggested that a mucosal enzyme is responsible. This paper reports a study intended to resolve this controversy. The hydrolysis of dietary phytic acid was measured in germfree and conventional rats fed either of two diets that differed in their calcium content. Ne...

  15. PLA recycling by hydrolysis at high temperature

    Cristina, Annesini Maria; Rosaria, Augelletti; Sara, Frattari; Fausto, Gironi


    In this work the process of PLA hydrolysis at high temperature was studied, in order to evaluate the possibility of chemical recycling of this polymer bio-based. In particular, the possibility to obtain the monomer of lactic acid from PLA degradation was investigated. The results of some preliminary tests, performed in a laboratory batch reactor at high temperature, are presented: the experimental results show that the complete degradation of PLA can be obtained in relatively low reaction times.

  16. Hydrolysis of ferric chloride in solution

    The Detox trademark process uses concentrated ferric chloride and small amounts of catalysts to oxidize organic compounds. It is under consideration for oxidizing transuranic organic wastes. Although the solution is reused extensively, at some point it will reach the acceptable limit of radioactivity or maximum solubility of the radioisotopes. This solution could be cemented, but the volume would be increased substantially because of the poor compatibility of chlorides and cement. A process has been developed that recovers the chloride ions as HCl and either minimizes the volume of radioactive waste or permits recycling of the radioactive chlorides. The process involves a two-step hydrolysis at atmospheric pressure, or preferably under a slight vacuum, and relatively low temperature, about 200 degrees C. During the first step of the process, hydrolysis occurs according to the reaction below: FeCl3liquid + H2O → FeOClsolid + 2 HClgas During the second step, the hot, solid, iron oxychloride is sprayed with water or placed in contact with steam, and hydrolysis proceeds to the iron oxide according to the following reaction: 2 FeOClsolid + H2O → Fe2O3solid + 2 HClgas. The iron oxide, which contains radioisotopes, can then be disposed of by cementation or encapsulation. Alternately, these chlorides can be washed off of the solids and can then either be recycled or disposed of in some other way

  17. Sugarcane bagasse hydrolysis using yeast cellulolytic enzymes.

    Souza, Angelica Cristina de; Carvalho, Fernanda Paula; Silva e Batista, Cristina Ferreira; Schwan, Rosane Freitas; Dias, Disney Ribeiro


    Ethanol fuel production from lignocellulosic biomass is emerging as one of the most important technologies for sustainable development. To use this biomass, it is necessary to circumvent the physical and chemical barriers presented by the cohesive combination of the main biomass components, which hinders the hydrolysis of cellulose and hemicellulose into fermentable sugars. This study evaluated the hydrolytic capacity of enzymes produced by yeasts, isolated from the soils of the Brazilian Cerrado biome (savannah) and the Amazon region, on sugarcane bagasse pre-treated with H2SO4. Among the 103 and 214 yeast isolates from the Minas Gerais Cerrado and the Amazon regions, 18 (17.47%) and 11 (5.14%) isolates, respectively, were cellulase-producing. Cryptococcus laurentii was prevalent and produced significant β- glucosidase levels, which were higher than the endo- and exoglucanase activities. In natura sugarcane bagasse was pre-treated with 2% H2SO4 for 30 min at 150oC. Subsequently, the obtained fibrous residue was subjected to hydrolysis using the Cryptococcus laurentii yeast enzyme extract for 72 h. This enzyme extract promoted the conversion of approximately 32% of the cellulose, of which 2.4% was glucose, after the enzymatic hydrolysis reaction, suggesting that C. laurentii is a good β-glucosidase producer. The results presented in this study highlight the importance of isolating microbial strains that produce enzymes of biotechnological interest, given their extensive application in biofuel production. PMID:23851270

  18. Palm Date Fibers: Analysis and Enzymatic Hydrolysis

    Mohammad J. Taherzadeh


    Full Text Available Waste palm dates were subjected to analysis for composition and enzymatic hydrolysis of their flesh fibers. The fruit contained 32% glucose and 30% fructose, while the water-insoluble fibers of its flesh consisted of 49.9% lignin and 20.9% polysaccharides. Water-insoluble fibers were settled to 55% of its initial volume in 12 h. The presence of skin and flesh colloidal fibers results in high viscosity and clogging problems during industrial processes. The settling velocity of the fibers was improved by enzymatic hydrolysis. Hydrolysis resulted in 84.3% conversion of the cellulosic part of the fibers as well as reducing the settling time to 10 minutes and the final settled volume to 4% of the initial volume. It implies easier separation of the fibers and facilitates fermentation processes in the corresponding industries. Two kinds of high- and low-lignin fibers were identified from the water-insoluble fibers. The high-lignin fibers (75% lignin settled easily, while the low-lignin fibers (41.4% lignin formed a slurry suspension which settled very slowly. The hydrophilicity of these low-lignin fibers is the major challenge of the industrial processes.

  19. Enzymatic Modification of Sphingomyelin

    Zhang, Long; Hellgren, Lars; Xu, Xuebing

    . Due to its major role in the water-retaining properties of the epidermis, ceramide is of great commercial potential in cosmetic and pharmaceuticals such as hair and skin care products. In current, ceramide is not easy to synthesis for industrial application and synthetic ceramide is still expensive...




    The involvement of the plasma membrane in the metabolism of the sphingolipids sphingomyelin (SM) and glucosylceramide (GlcCer) was studied, employing fluorescent short-chain analogues of these lipids, 6-[N-(7-nitro-2,1,3-benzoxadiazol-4-yl)amino]hexanoylsphingosylphosphorylcholine (C-6-NBD-SM), C-6-

  1. Effects of microtubule mechanics on hydrolysis and catastrophes

    We introduce a model for microtubule (MT) mechanics containing lateral bonds between dimers in neighboring protofilaments, bending rigidity of dimers, and repulsive interactions between protofilaments modeling steric constraints to investigate the influence of mechanical forces on hydrolysis and catastrophes. We use the allosteric dimer model, where tubulin dimers are characterized by an equilibrium bending angle, which changes from 0∘ to 22∘ by hydrolysis of a dimer. This also affects the lateral interaction and bending energies and, thus, the mechanical equilibrium state of the MT. As hydrolysis gives rise to conformational changes in dimers, mechanical forces also influence the hydrolysis rates by mechanical energy changes modulating the hydrolysis rate. The interaction via the MT mechanics then gives rise to correlation effects in the hydrolysis dynamics, which have not been taken into account before. Assuming a dominant influence of mechanical energies on hydrolysis rates, we investigate the most probable hydrolysis pathways both for vectorial and random hydrolysis. Investigating the stability with respect to lateral bond rupture, we identify initiation configurations for catastrophes along the hydrolysis pathways and values for a lateral bond rupture force. If we allow for rupturing of lateral bonds between dimers in neighboring protofilaments above this threshold force, our model exhibits avalanche-like catastrophe events. (papers)

  2. Hydrolysis of ferric chloride in solution

    Lussiez, G.; Beckstead, L.


    The Detox{trademark} process uses concentrated ferric chloride and small amounts of catalysts to oxidize organic compounds. It is under consideration for oxidizing transuranic organic wastes. Although the solution is reused extensively, at some point it will reach the acceptable limit of radioactivity or maximum solubility of the radioisotopes. This solution could be cemented, but the volume would be increased substantially because of the poor compatibility of chlorides and cement. A process has been developed that recovers the chloride ions as HCl and either minimizes the volume of radioactive waste or permits recycling of the radioactive chlorides. The process involves a two-step hydrolysis at atmospheric pressure, or preferably under a slight vacuum, and relatively low temperature, about 200{degrees}C. During the first step of the process, hydrolysis occurs according to the reaction below: FeCl{sub 3 liquid} + H{sub 2}O {r_arrow} FeOCl{sub solid} + 2 HCl{sub gas} During the second step, the hot, solid, iron oxychloride is sprayed with water or placed in contact with steam, and hydrolysis proceeds to the iron oxide according to the following reaction: 2 FeOCl{sub solid} + H{sub 2}O {r_arrow} Fe{sub 2}O{sub 3 solid} + 2 HCl{sub gas}. The iron oxide, which contains radioisotopes, can then be disposed of by cementation or encapsulation. Alternately, these chlorides can be washed off of the solids and can then either be recycled or disposed of in some other way.

  3. Hydrolysis of Fish Protein by Analkaline Protease


    Cod muscle protein was hydrolyzed by an alkaline protease in our study. The influences of hydrolysis temperature,fish protein concentration,and ratio of protease addition to protein amount on its degree of hy drolysis (DH) of protein were studied in details by applying dual quadratic rotary combinational design. The final results showed that more than 84% cod muscle protein could be hydrolyzed and recovered. Cod protein hydrolysate thus obtained had a balanced amino acid composition and mainly consisted of small peptides with molecule weight less than 6900 dalton.

  4. Pretreatment and enzymatic hydrolysis of lignocellulosic biomass

    Corredor, Deisy Y.

    The performance of soybean hulls and forage sorghum as feedstocks for ethanol production was studied. The main goal of this research was to increase fermentable sugars' yield through high-efficiency pretreatment technology. Soybean hulls are a potential feedstock for production of bio-ethanol due to their high carbohydrate content (≈50%) of nearly 37% cellulose. Soybean hulls could be the ideal feedstock for fuel ethanol production, because they are abundant and require no special harvesting and additional transportation costs as they are already in the plant. Dilute acid and modified steam-explosion were used as pretreatment technologies to increase fermentable sugars yields. Effects of reaction time, temperature, acid concentration and type of acid on hydrolysis of hemicellulose in soybean hulls and total sugar yields were studied. Optimum pretreatment parameters and enzymatic hydrolysis conditions for converting soybean hulls into fermentable sugars were identified. The combination of acid (H2SO4, 2% w/v) and steam (140°C, 30 min) efficiently solubilized the hemicellulose, giving a pentose yield of 96%. Sorghum is a tropical grass grown primarily in semiarid and dry parts of the world, especially in areas too dry for corn. The production of sorghum results in about 30 million tons of byproducts mainly composed of cellulose, hemicellulose, and lignin. Forage sorghum such as brown midrib (BMR) sorghum for ethanol production has generated much interest since this trait is characterized genetically by lower lignin concentrations in the plant compared with conventional types. Three varieties of forage sorghum and one variety of regular sorghum were characterized and evaluated as feedstock for fermentable sugar production. Fourier transform infrared spectroscopy (FTIR), scanning electron microscope (SEM) and X-Ray diffraction were used to determine changes in structure and chemical composition of forage sorghum before and after pretreatment and enzymatic hydrolysis

  5. Palm Date Fibers: Analysis and Enzymatic Hydrolysis

    Taherzadeh, Mohammad J.; Keikhosro Karimi; Marzieh Shafiei


    Waste palm dates were subjected to analysis for composition and enzymatic hydrolysis of their flesh fibers. The fruit contained 32% glucose and 30% fructose, while the water-insoluble fibers of its flesh consisted of 49.9% lignin and 20.9% polysaccharides. Water-insoluble fibers were settled to 55% of its initial volume in 12 h. The presence of skin and flesh colloidal fibers results in high viscosity and clogging problems during industrial processes. The settling velocity of the fibers was i...

  6. FTIR-ATR study of the influence of the pyrimidine analog of fluphenazine on the chain-melting phase transition of sphingomyelin membranes

    Kuć, Marta; Cieślik-Boczula, Katarzyna; Świątek, Piotr; Jaszczyszyn, Agata; Gąsiorowski, Kazimierz; Malinka, Wiesław


    The membrane perturbing potency of the highly effective anti-multidrug resistance (MDR) pyrimidine analog of fluphenazine (FPh-prm), has been studied using attenuated total reflectance Fourier-transfer infrared spectroscopy (FTIR-ATR). The temperature- and FPh-prm-dose-dependent evolutions of the infrared spectra of FPh-prm/sphingomyelin (SM) mixtures were analyzed using principal component analysis (PCA). It has been postulated that the distinct anti-MDR activity of FPh-prm could be related to its ability to affect the modification of SM membranes. A reduction in the temperature of the chain-melting phase transition was observed in FPh-prm-mixed SM membranes together with the loosing of the phase transition cooperativity. Increasing the temperature led to the trans to gauche isomerization of FPh-prm-rich lipid membranes, which resulted in the gradual release of FPh-prm from the lipid membrane to the water phase.

  7. Enzymatic Hydrolysis of Various Proteins of Wheat in Heterogeneous Conditions

    Hasan Hasanov


    Full Text Available Enzymatic hydrolysis of different proteins isolated from wheat flour by neutral proteinase (neutraza "Novozymes" was studied. It was shown, that hydrolysis of alkaline proteins was 10-11 times higher as compared with albumin from wheat, 3-4 times higher than alcohol soluble proteins and 2-2.5 times higher than globulins. It was found that, hydrothermal treatment of wheat flour decreased the rate of protein hydrolysis. The rate of hydrolysis of native alkaline soluble proteins was 4-6 times higher than denaturized proteins. The rate of hydrolysis of denatured water-soluble proteins is 3-5 times higher as compared with native protein (albumin. It was shown that product of thermal degradation of raw materials also influence on the rate of protein hydrolysis.

  8. Water Availability as a Measure of Cellulose Hydrolysis Efficiency

    Hsieh, Chia-Wen

    Enzymatic hydrolysis involves the use of cellulases to break down cellulose in the presence of water. Therefore, not only are enzyme and substrate properties important for efficient hydrolysis, but also the hydrolysis medium, i.e. the liquid phase. The LF-NMR technique is used in this work to...... measure properties of the liquid phase, where water protons are characterized based on their mobility in the system as measured by their relaxation time. Studies of cellulose hydrolysis at low dry matter show that the contents of the liquid phase influence the final hydrolysis yield, as the presence of...... sugars, salts, and surfactants impact the water relaxation time. Systems with high concentrations of sugars and salts tend to have low water availability, as these form strong interactions with water to keep their solubility, leaving less water available for hydrolysis. Thus, cellulase performance...

  9. The Hydrolysis of Carbonyl Sulfide at Low Temperature: A Review

    Zhao, Shunzheng; Yi, Honghong; Tang, Xiaolong; Jiang, Shanxue; Gao, Fengyu; Zhang, Bowen; Zuo, Yanran; Wang, Zhixiang


    Catalytic hydrolysis technology of carbonyl sulfide (COS) at low temperature was reviewed, including the development of catalysts, reaction kinetics, and reaction mechanism of COS hydrolysis. It was indicated that the catalysts are mainly involved metal oxide and activated carbon. The active ingredients which can load on COS hydrolysis catalyst include alkali metal, alkaline earth metal, transition metal oxides, rare earth metal oxides, mixed metal oxides, and nanometal oxides. The catalytic hydrolysis of COS is a first-order reaction with respect to carbonyl sulfide, while the reaction order of water changes as the reaction conditions change. The controlling steps are also different because the reaction conditions such as concentration of carbonyl sulfide, reaction temperature, water-air ratio, and reaction atmosphere are different. The hydrolysis of carbonyl sulfide is base-catalyzed reaction, and the force of the base site has an important effect on the hydrolysis of carbonyl sulfide. PMID:23956697

  10. [Functional state of a sphingomyeline cycle and free radical lipid oxidation activity of a rat's liver during different phases of starvation].

    Kuz'menko, D I; Burov, P G; Serebrov, V Iu; Faĭt, E A; Perevozchikova, T V


    The functional state of a sphingomyeline cycle and character of its mutual relations with the processes of free radical lipid oxidation during starvation of animals without any restriction of access to drinking water at 1, 2, 3 day (I phase) and 6 day (II phase of starvation) were studied at the liver of rats. The maximal values of the ceramide/sphingomyeline ratio and activity neutral sphingomyelinase and executive caspase-3 were reached in a liver of animals at the 3rd day of starvation. From the 3rd day of starvation the concentration of the tumour necrosis factor-alpha which is one of activators neutral sphingomyelinase was increase in rats blood serum. During the extent of large part of the I phase of starvation the intensity of free radical lipid peroxidation in a liver had almost the same level as in control group--that was a result of the high-grade functioning of antioxidant defense system. After transition the I phase of starvation into the II phase (6 day of experiment) the oxidative stress was developed as result of an exhaustion of system antioxidant defense potential in a liver. The results of this data can testify that during I phase of starvation in a liver the conditions was raised for display of the ceramide-mediated proapoptotic signalling. We assume that ceramide-mediated apoptosis is one of mechanisms of optimization of liver cellular population at the frames of metabolic adaptation. The I phase of starvation in a liver proves by the ceramide-mediated proapoptotic signaling developing. During the II phase of starvation the oxidative stress process were prevailed. PMID:23289297

  11. Mechanisms and kinetics of the hydrolysis and condensation of alkoxides

    Schmidt, Helmut K.; Scholze, Horst


    Hydrolysis and condensation of alkoxides involve different reaction steps. Generally the first step is the dissolution of monomers in organic solvents like alcohols. The second step is hydrolysis, where in most cases condensation may not be separated. Dissolution may incorporate solvatation, coordination, complexation of polymerization. The addition of water leads to hydrolysis of Si-O-C bonds and subsequently condensation of silanoles takes place. Another possibility of reaction is the hydro...

  12. Hydrolysis of sunflower proto pectin in static and dynamic mode

    The article describes a hydrolysis process of sunflower head residue by using state and dynamics regime of hydrolysis. It's shown that application of dynamics method positively influences on the pectin yields and its main parameters. The results of comparative study of acid concentration effect on pectin parameters in two different hydrolysis mode allows to develop a more effective process in the pectin production. (author)

  13. Hydrolysis-extraction of apple proto pectins in dynamic mode

    The article describes a hydrolysis process of apple husks by using dynamics regime of hydrolysis. It's shown that application of dynamics method positively influences on the pectin yields and its main parameters. It was defined that by dynamics regime of hydrolysis-extraction of apple husks it is possible to obtain qualitative products with high yield at a mild ph value of medium of hydrolysing agent.

  14. Muscarinic receptor activation of phosphatidylcholine hydrolysis. Relationship to phosphoinositide hydrolysis and diacylglycerol metabolism

    Martinson, E.A.; Goldstein, D.; Brown, J.H. (Univ. of California, San Diego, La Jolla (USA))


    We examined the relationship between phosphatidylcholine (PC) hydrolysis, phosphoinositide hydrolysis, and diacylglycerol (DAG) formation in response to muscarinic acetylcholine receptor (mAChR) stimulation in 1321N1 astrocytoma cells. Carbachol increases the release of (3H)choline and (3H)phosphorylcholine ((3H)Pchol) from cells containing (3H)choline-labeled PC. The production of Pchol is rapid and transient, while choline production continues for at least 30 min. mAChR-stimulated release of Pchol is reduced in cells that have been depleted of intracellular Ca2+ stores by ionomycin pretreatment, whereas choline release is unaffected by this pretreatment. Phorbol 12-myristate 13-acetate (PMA) increases the release of choline, but not Pchol, from 1321N1 cells, and down-regulation of protein kinase C blocks the ability of carbachol to stimulate choline production. Taken together, these results suggest that Ca2+ mobilization is involved in mAChR-mediated hydrolysis of PC by a phospholipase C, whereas protein kinase C activation is required for mAChR-stimulated hydrolysis of PC by a phospholipase D. Both carbachol and PMA rapidly increase the formation of (3H)phosphatidic acid ((3H)PA) in cells containing (3H)myristate-labeled PC. (3H)Diacylglycerol ((3H)DAG) levels increase more slowly, suggesting that the predominant pathway for PC hydrolysis is via phospholipase D. When cells are labeled with (3H)myristate and (14C)arachidonate such that there is a much greater 3H/14C ratio in PC compared with the phosphoinositides, the 3H/14C ratio in DAG and PA increases with PMA treatment but decreases in response to carbachol.

  15. Dynamic modeling and validation of a lignocellulosic enzymatic hydrolysis process

    Prunescu, Remus Mihail; Sin, Gürkan


    The enzymatic hydrolysis process is one of the key steps in second generation biofuel production. After being thermally pretreated, the lignocellulosic material is liquefied by enzymes prior to fermentation. The scope of this paper is to evaluate a dynamic model of the hydrolysis process on a...... demonstration scale reactor. The following novel features are included: the application of the Convection–Diffusion–Reaction equation to a hydrolysis reactor to assess transport and mixing effects; the extension of a competitive kinetic model with enzymatic pH dependency and hemicellulose hydrolysis; a...

  16. Acid-functionalized nanoparticles for biomass hydrolysis

    Pena Duque, Leidy Eugenia

    Cellulosic ethanol is a renewable source of energy. Lignocellulosic biomass is a complex material composed mainly of cellulose, hemicellulose, and lignin. Biomass pretreatment is a required step to make sugar polymers liable to hydrolysis. Mineral acids are commonly used for biomass pretreatment. Using acid catalysts that can be recovered and reused could make the process economically more attractive. The overall goal of this dissertation is the development of a recyclable nanocatalyst for the hydrolysis of biomass sugars. Cobalt iron oxide nanoparticles (CoFe2O4) were synthesized to provide a magnetic core that could be separated from reaction using a magnetic field and modified to carry acid functional groups. X-ray diffraction (XRD) confirmed the crystal structure was that of cobalt spinel ferrite. CoFe2O4 were covered with silica which served as linker for the acid functions. Silica-coated nanoparticles were functionalized with three different acid functions: perfluoropropyl-sulfonic acid, carboxylic acid, and propyl-sulfonic acid. Transmission electron microscope (TEM) images were analyzed to obtain particle size distributions of the nanoparticles. Total carbon, nitrogen, and sulfur were quantified using an elemental analyzer. Fourier transform infra-red spectra confirmed the presence of sulfonic and carboxylic acid functions and ion-exchange titrations accounted for the total amount of catalytic acid sites per nanoparticle mass. These nanoparticles were evaluated for their performance to hydrolyze the beta-1,4 glycosidic bond of the cellobiose molecule. Propyl-sulfonic (PS) and perfluoropropyl-sulfonic (PFS) acid functionalized nanoparticles catalyzed the hydrolysis of cellobiose significantly better than the control. PS and PFS were also evaluated for their capacity to solubilize wheat straw hemicelluloses and performed better than the control. Although PFS nanoparticles were stronger acid catalysts, the acid functions leached out of the nanoparticle during

  17. Hydrolysis and formation constants at 250C

    A database consisting of hydrolysis and formation constants for about 20 metals associated with the disposal of nuclear waste is given. Complexing ligands for the various ionic species of these metals include OH, F, Cl, SO4, PO4 and CO3. Table 1 consists of tabulated calculated and experimental values of log K/sub xy/, mainly at 250C and various ionic strengths together with references to the origin of the data. Table 2 consists of a column of recommended stability constants at 250C and zero ionic strength tabulated in the column headed log K/sub xy/(0); other columns contain coefficients for an extended Debye-Huckel equation to permit calculations of stability constants up to 3 ionic strength, and up to 0.7 ionic strength using the Davies equation. Selected stability constants calculated with these coefficients for various ionic strengths agree to an average of +- 2% when compared with published experimental and calculated values

  18. Pretreatment and enzymatic hydrolysis of corn fiber

    Grohmann, K. [USDA Citrus and Subtropical Products Research Labs., Winter Haven, CT (United States); Bothast, R.J. [National Center for Agricultural Utilization Research, Peoria, IL (United States)


    Corn fiber is a co-product of the corn wet milling industry which is usually marketed as a low value animal feed ingredient. Approximately 1.2 x 10{sup 6} dry tons of this material are produced annually in the United States. The fiber is composed of kernel cell wall fractions and a residual starch which can all be potentially hydrolyzed to a mixture of glucose, xylose, arabinose and galactose. We have investigated a sequential saccharification of polysaccharides in corn fiber by a treatment with dilute sulfuric acid at 100 to 160{degrees}C followed by partial neutralization and enzymatic hydrolysis with mixed cellulose and amyloglucosidase enzymes at 45{degrees}C. The sequential treatment achieved a high (approximately 85%) conversion of all polysaccharides in the corn fiber to monomeric sugars, which were in most cases fermentable to ethanol by the recombinant bacterium Escherichia coli KOll.

  19. Enzymatic hydrolysis of PTT polymers and oligomers.

    Eberl, A; Heumann, S; Kotek, R; Kaufmann, F; Mitsche, S; Cavaco-Paulo, A; Gübitz, G M


    Oligomers and polymers (film, fabrics) of the linear aromatic polyester poly(trimethylene terephthalate) (PTT) were treated with polyesterases from Thermomyces lanuginosus, Penicillium citrinum, Thermobifida fusca and Fusarium solani pisi. The cutinase from T. fusca was found to release the highest amounts of hydrolysis products from PTT materials and was able to open and hydrolyse a cyclic PTT dimer according to RP-HPLC-UV detection. In contrast, the lipase from T. lanuginosus also showed activity on the PTT fibres and on bis(3-hydroxypropyl) terephthalate (BHPT) but was not able to hydrolyse the polymer film, mono(3-hydroxypropyl) terephthalate (MHPT) nor the cyclic dimer of PTT. As control enzymes inhibited with mercury chloride were used. Surface hydrophilicity changes were investigated with contact angle measurements and the degree of crystallinity changes were determined with DSC. PMID:18405994

  20. Microtubule dynamics: Caps, catastrophes, and coupled hydrolysis

    Flyvbjerg, H.; Holy, T.E.; Leibler, S.


    probability distributions relating to available experimental data are derived. Caps are found to be short and the total rate of hydrolysis at a microtubule end is found to be dynamically coupled to growth. The so-called catastrophe rate is a simple function of the microtubule growth rare and fits experimental...... data. A constant nonzero catastrophe rare, identical for both microtubule ends, is predicted at large growth rates. The delay time for dilution-induced catastrophes is stochastic with a simple distribution that fits the experimental one and, like the experimental one, does not depend on the rate of...... unified description of several apparently contradictory experimental data. Experimental results for the catastrophe rate at different concentrations of magnesium ions and of microtubule associated proteins are discussed in terms of the model. Feasible experiments are suggested that can provide decisive...

  1. Bioabatement with hemicellulase supplementation to reduce enzymatic hydrolysis inhibitors

    Removal of inhibitory compounds by bioabatement, combined with xylan hydrolysis, enables effective cellulose hydrolysis of pretreated corn stover, for fermentation of the sugars to fuel ethanol or other products. The fungus Coniochaeta ligniaria NRRL30616 eliminates most enzyme and fermentation inhi...

  2. Bioethanol from lignocellulose - pretreatment, enzyme immobilization and hydrolysis kinetics

    Tsai, Chien Tai

    Pretreatment and enzymatic hydrolysis are two of the processes involved in the production of cellulosic ethanol. Several pretreatment methods were proposed, however new pretreatment strategies to increase enzymetic hydrolysis efficiency are still under investigation. For enzymatic hydrolysis, the...... economic considerations, barley straw can be pretreated under 150°C for 50 min with dry matter of 20% (w/w). Glucose yield can be up to 70% after enzymatic hydrolysis. (2) Immobilization of ß-glucosidase (BG), which was done during 2010. One of the major bottlenecks in production of ethanol from...... different product inhibitors such as glucose, cellobiose and xylose) to test the hydrolysis and product inhibition mechanism of the model. Nonlinear least squares methodwas used to identify the model and estimate kinetic parameters based on the experimental data. The analysis showed that transglycosylation...

  3. Kinetics of enzymatic hydrolysis of methyl ricinoleate

    Neeharika, T. S.V.R.


    Full Text Available Ricinoleic acid is an unsaturated hydroxy fatty acid that naturally occurs in castor oil in proportions of up to 85–90%. Ricinoleic acid is a potential raw material and finds several applications in coatings, lubricant formulations and pharmaceutical areas. Enzymatic hydrolysis of castor oil is preferred over conventional hydrolysis for the preparation of ricinoleic acid to avoid estolide formation. A kinetics analysis of the enzymatic hydrolysis of Methyl Ricinoleate in the presence of Candida antarctica Lipase B was carried out in this study by varying reaction temperature (40–60 °C and enzyme concentration (2–5%. The optimal conditions were found to be 6 h reaction time, temperature 60°C, buffer to methyl ricinoleate ratio 2:1(v/w and 4% enzyme concentration to achieve a maximum conversion of 98.5%. A first order reversible reaction kinetic model was proposed to describe this reaction and a good agreement was observed between the experimental data and the model values. The effect of temperature on the forward reaction rate constant was determined by fitting data to the Arrhenius equation. The activation energy for forward reaction was found to be 14.69 KJ·mol−1.El ácido ricinoleico es un hidroxiácido insaturado que se produce naturalmente en el aceite de ricino en proporciones de hasta el 85–90%. El ácido ricinoleico es una materia prima con gran potencial y tiene aplicaciones en revestimientos, formulaciones lubricantes y en áreas farmacéuticas. Para la preparación del ácido ricinoleico se prefiere la hidrólisis enzimática del aceite de ricino a la hidrólisis convencional, para evitar la formación de estólidos. En este estudio se llevó a cabo la cinética de la hidrólisis enzimática del ricinoleato de metilo en presencia de lipasa de Candida antarctica B mediante la variación de la temperatura de reacción (40–60 °C y la concentración de la enzima (2–5%. Las condiciones óptimas de la reacción para

  4. Enzyme loading dependence of cellulose hydrolysis of sugarcane bagasse

    Carlos Martín


    Full Text Available The enzymatic hydrolysis of steam-pretreated sugarcane bagasse, either delignified or non-delignified, was studied as a function of enzyme loading. Hydrolysis experiments were carried out using five enzyme loadings (2.5 to 20 FPU/g cellulose and the concentration of solids was 2% for both materials. Alkaline delignification improved cellulose hydrolysis by increasing surface area. For both materials, glucose concentrations increased with enzyme loading. On the other hand, enzyme loadings higher than 15 FPU/g did not result in any increase in the initial rate, since the excess of enzyme adsorbed onto the substrate restricted the diffusion process through the structure.

  5. Furfural production from fruit shells by acid-catalyzed hydrolysis

    Demirbas, A. [Selcuk Univ., Konya (Turkey). Dept. of Chemical Engineering


    Pentosans are hydrolyzed to pentoses by dilute mineral acid hydrolysis. The main source of pentosans is hemicelluloses. Furfural can be produced by the acid hydrolysis of pentosan from fruit shells such as hazelnut, sunflower, walnut, and almond of agricultural wastes. Further dehydration reactions of the pentoses yield furfural. The hydrolysis of each shell sample was carried out in dilute sulfuric acid (0.05 to 0.200 mol/l), at high temperature (450-525 K), and short reaction times (from 30 to 600 s). (author)

  6. Hydrolysis of xenon difluoride in acetonitride-water mixtures

    Hydrolysis of xenon difluoride in acetonitride-water mixture and at XeF2 concentrations 0.03-0.100 mol/l has been studied. It is sown, that the hydrolysis is catalyzed by hydrogen fluoride, formed as a result of interaction between water and XeF2. It is ascertained, that half-life of xenon difluoride decreases with the temperature increase from 30 to 50 deg C and water concentration from 0.5 to 22.2 mol/l. Hydrolysis mechanism is discussed


    Jie Lu; Shulan Shi; Runan Yang; Fuzheng Liang


    The optimum conditions of hydrolysis of cellulosic wastes by cellulase were studied. The results show that the optimum conditions of sulfuric acid pretreatment were sulfuric acid consistency 0.3M,pretreatment temperature 100℃, pretreatment time 4hours. After sulfuric acid pretreatment, the optimum conditions of hydrolysis by cellulase were enzymatic temperature 50℃ ,enzymatic time 48hours,pH4.8,the charge of cellulase 100IU/g and the substraste consistency 60g/l. Meanwhile this paper studies that the structural change of cellulose during sulfuric acid pretreatment and cellulase hydrolysis by analyzing the infrared spectra.


    JieLu; ShulanShi; RunanYang; FuzhengLiang


    The optimum conditions of hydrolysis of cellulosic wastes by cellulase were studied. The results show that the optimum conditions of sulfuric acid pretreatment were sulfuric acid consistency 0.3M, pretreatment temperature 100℃, pretreatment time 4hours. After sulfuric acid pretreatment, the optimum conditions of hydrolysis by cellulase were enzymatic temperature 50℃, enzymatic time 48hours,pH4.8,the charge of cellulase 100IU/g and the substraste consistency 60g/l. Meanwhile this paper studies that the structural change of cellulose during sulfuric acid pretreatment and cellulase hydrolysis by analyzing the infrared spectra.

  9. Improvement of fat enzymatic hydrolysis technology

    Некрасов, Павло Олександрович; Плахотна, Юлія Миколаївна; Некрасов, Олександр Павлович


    Дана робота присвячена дослідженню гідролізу олії під дією ферментних препаратів вітчизняного та закордонного виробництва. Створені математичні моделі процесів та оптимізовано їх параметри. This article covers the investigation of hydrolysis of oil by domestic and foreign produced enzymes. The mathematical models of the processes were designed and process conditions were optimized....

  10. Enzymatic hydrolysis of biomass from wood.

    Álvarez, Consolación; Reyes-Sosa, Francisco Manuel; Díez, Bruno


    Current research and development in cellulosic ethanol production has been focused mainly on agricultural residues and dedicated energy crops such as corn stover and switchgrass; however, woody biomass remains a very important feedstock for ethanol production. The precise composition of hemicellulose in the wood is strongly dependent on the plant species, therefore different types of enzymes are needed based on hemicellulose complexity and type of pretreatment. In general, hardwood species have much lower recalcitrance to enzymes than softwood. For hardwood, xylanases, beta-xylosidases and xyloglucanases are the main hemicellulases involved in degradation of the hemicellulose backbone, while for softwood the effect of mannanases and beta-mannosidases is more relevant. Furthermore, there are different key accessory enzymes involved in removing the hemicellulosic fraction and increasing accessibility of cellulases to the cellulose fibres improving the hydrolysis process. A diversity of enzymatic cocktails has been tested using from low to high densities of biomass (2-20% total solids) and a broad range of results has been obtained. The performance of recently developed commercial cocktails on hardwoods and softwoods will enable a further step for the commercialization of fuel ethanol from wood. PMID:26833542

  11. Full scale enzymatic hydrolysis; Enzymhydrolyse i fuldskala


    An investigation of the feasibility of adding enzymes during the process of anaerobic biomass conversion in order to increase the production of methane is described. The experiment was carried out at a biomass conversion plant at Lintrup (Denmark) in order to compare field results with a laboratory experiment of the same nature. During the continuous fermentation, direct doses of a relatively large amount of enzymes (ca. 6 ml enzyme concentrate mixture per kg dry matter) were added. The idea was that in this way an increase in the amount of hydrolyzed material during a short period could be achieved and thus the conversion of matter that is otherwise difficult to decompose (in this case slurried manures and organic wastes from the food and medicinal industries) could be optimized. This would result in an increased growth of the microorganisms, especially the hydrolytic fermentative. The increase in the growth of the bacteria results in an increase in the amount of extracellular enzymes produced by the hydrolytic fermentative bacteria, and this should promote the hydrolysis of the polymeric material. It was found that the use of enzymes as additives did not increase the production of methane in the anaerobic biomass conversion plant. (AB)

  12. Evaluation of hydrolysis-esterification biodiesel production from wet microalgae.

    Song, Chunfeng; Liu, Qingling; Ji, Na; Deng, Shuai; Zhao, Jun; Li, Shuhong; Kitamura, Yutaka


    Wet microalgae hydrolysis-esterification route has the advantage to avoid the energy-intensive units (e.g. drying and lipid extraction) in the biodiesel production process. In this study, techno-economic evaluation of hydrolysis-esterification biodiesel production process was carried out and compared with conventional (usually including drying, lipid extraction, esterification and transesterification) biodiesel production process. Energy and material balance of the conventional and hydrolysis-esterification processes was evaluated by Aspen Plus. The simulation results indicated that drying (2.36MJ/L biodiesel) and triolein transesterification (1.89MJ/L biodiesel) are the dominant energy-intensive stages in the conventional route (5.42MJ/L biodiesel). By contrast, the total energy consumption of hydrolysis-esterification route can be reduced to 1.81MJ/L biodiesel, and approximately 3.61MJ can be saved to produce per liter biodiesel. PMID:27209457

  13. Total decomposition of organophosphate pesticides by microwave-assisted hydrolysis

    Čechová, Lucie; Jansa, Petr; Dračínský, Martin; Janeba, Zlatko

    Lanzarote: -, 2012. s. 69-69. [Zing Conferences: Microwave and Flow Conference. 28.02.2012-02.03.2012, Lanzarote] Institutional research plan: CEZ:AV0Z40550506 Keywords : microwave * hydrolysis * organophosphates * pesticides Subject RIV: CC - Organic Chemistry

  14. Improved Method for Isolation of Bacterial Inhibitors from Oleuropein Hydrolysis

    Federici, Federico; Bongi, Guido


    A new high-pressure liquid chromatography multidetection quantitative method for the isolation of the products of oleuropein hydrolysis is described. A single analysis yields sufficient amounts of the compounds to test their inhibitory effect on bacterial growth.

  15. Enzymatic hydrolysis of steryl glycosides for their analysis in foods.

    Münger, Linda H; Nyström, Laura


    Steryl glycosides (SG) contribute significantly to the total intake of phytosterols. The standard analytical procedure involving acid hydrolysis fails to reflect the correct sterol profile of SG due to isomerization of some of the labile sterols. Therefore, various glycosylases were evaluated for their ability to hydrolyse SG under milder conditions. Using a pure SG mixture in aqueous solution, the highest glycolytic activity, as demonstrated by the decrease in SG and increase in free sterols was achieved using inulinase preparations (decrease of >95%). High glycolytic activity was also demonstrated using hemicellulase (63%). The applicability of enzymatic hydrolysis using inulinase preparations was further verified on SG extracted from foods. For example in potato peel Δ(5)-avenasteryl glucoside, a labile SG, was well preserved and contributed 26.9% of the total SG. Therefore, enzymatic hydrolysis is suitable for replacing acid hydrolysis of SG in food lipid extracts to accurately determine the sterol profile of SG. PMID:24912717

  16. Mechanism of cellobiose inhibition in cellulose hydrolysis by cellobiohydrolase

    ZHAO; Yue; WU; Bin; YAN; Baixu; GAO; Peiji


    An experimental study of cellobiose inhibition in cellulose hydrolysis by synergism of cellobiohydrolyse I and endoglucanase I is presented. Cellobiose is the structural unit of cellulose molecules and also the main product in enzymatic hydrolysis of cellulose. It has been identified that cellobiose can strongly inhibit hydrolysis reaction of cellulase, whereas it has no effect on the adsorption of cellulase on cellulose surface. The experimental data of FT-IR spectra, fluorescence spectrum and circular dichroism suggested that cellobiose can be combined with tryptophan residue located near the active site of cellobiohydrolase and then form steric hindrance, which prevents cellulose molecule chains from diffusing into active site of cellulase. In addition, the molecular conformation of cellobiohydrolase changes after cellobiose binding, which also causes most of the non-productive adsorption. Under these conditions, microfibrils cannot be separated from cellulose chains, thus further hydrolysis of cellulose can hardly proceed.

  17. Enzymatic hydrolysis of pretreated barley and wheat straw

    Rosgaard, Lisa


    work involved evaluation of 1) possible ways to increase the glucose release from the commercial cellulase product Celluclast by boosting with other enzyme activities to increase the enzymatic hydrolysis, 2) comparing differently pretreated feedstock substrates and 3) evaluating a fed-batch substrate...... feeding strategy to increase the substrate loading in the hydrolysis reaction. The substrate for the enzymatic hydrolysis was primarily steam pretreated wheat and barley straw since these substrates were the primary feedstocks for the Babilafuente Bioethanol process. The initial work showed that there was...... different pretreatment conditions; hot water extraction and acid- or water impregnation followed by steam explosion showed there were slight differences between the effect of pretreatment conditions in relation to the overall yield from enzymatic hydrolysis. The highest glucose concentration was found for...

  18. Evaluation of wet oxidation pretreatment for enzymatic hydrolysis of softwood

    Palonen, H.; Thomsen, A.B.; Tenkanen, M.;


    The wet oxidation pretreatment (water, oxygen, elevated temperature, and pressure) of softwood (Picea abies) was investigated for enhancing enzymatic hydrolysis. The pretreatment was preliminarily optimized. Six different combinations of reaction time, temperature, and pH were applied, and the...... compositions of solid and liquid fractions were analyzed. The solid fraction after wet oxidation contained 58-64% cellulose, 2-16% hemicellulose, and 24-30% lignin. The pretreatment series gave information about the roles of lignin and hemicellulose in the enzymatic hydrolysis. The temperature of the...... pretreatment, the residual hemicellulose content of the substrate, and the type of the commercial cellulase preparation used were the most important factors affecting the enzymatic hydrolysis. The highest sugar yield in a 72-h hydrolysis, 79% of theoretical, was obtained using a pretreatment of 200degreesC for...

  19. Analysis of myo-inositol hexakisphosphate hydrolysis by Bacillus phytase

    Kerovuo, J.; Rouvinen, J.; Hatzack, Frank-Andreas


    Phytic acid (myo-inositol hexakisphosphate, InsP(6)) hydrolysis by Bacillus phytase (PhyC) was studied. The enzyme hydrolyses only three phosphates from phytic acid. Moreover, the enzyme seems to prefer the hydrolysis of every second phosphate over that of adjacent ones. Furthermore, it is very...... likely that the enzyme has two alternative pathways for the hydrolysis of phytic acid, resulting in two different myo-inositol trisphosphate end products: Ins(2,4,6)P-8 and Ins(1,3,5)P-3. These results, together with inhibition studies with fluoride, vanadate, substrate and a substrate analogue, indicate......) computer-modelling analyses of enzyme-substrate complexes, a novel mode of phytic acid hydrolysis is proposed....

  20. Characterization of casein hydrolysates derived from enzymatic hydrolysis

    Wang, Jinshui; Su, Yinjie; Jia, Feng; Jin, Huali


    Background Casein is the main proteinaceous component of milk and has made us interest due to its wide applications in the food, drug, and cosmetic industries as well as to its importance as an investigation material for elucidating essential questions regarding the protein chemistry. Enzymatic hydrolysis is an important method commonly used in the modification of protein structure in order to enhance the functional properties of proteins. The relationship between enzymatic hydrolysis and str...

  1. Hydrolysis of starch by sorghum malt for maltodextrin production [abstract

    Thonart, P.; Destain, J.; Ba, K.


    Maltodextrin is a mixture of saccharides with a molecular weight between polysaccharides and oligosaccharides with DE lower than 20. Maltodextrin is more soluble in water than native starches, also is cheaper in comparison with other major edible hydrocolloids. Maltodextrin is obtained by moderate enzymatic or acidic hydrolysis of starch. The hydrolysis of starch, catalyzed by amylases, is the most important commercial enzyme process. The hydrolyzed products are widely applied in food, paper ...

  2. Hydrolysis of starch by sorghum malt for maltodextrin production

    Ba, K.; Destain, Jacqueline; Thonart, Philippe


    Maltodextrin is a mixture of saccharides with a molecular weight between polysaccharides and oligosaccharides with DE lower than 20. Maltodextrin is more soluble in water than native starches, also is cheaper in comparison with other major edible hydrocolloids. Maltodextrin is obtained by moderate enzymatic or acidic hydrolysis of starch. The hydrolysis of starch, catalyzed by amylases, is the most important commercial enzyme process. The hydrolyzed products are widely applied in food, paper ...

  3. DNA-Catalyzed Hydrolysis of Esters and Aromatic Amides

    Brandsen, Benjamin M.; Hesser, Anthony R.; Castner, Marissa A.; Chandra, Madhavaiah; Silverman, Scott K.


    We previously reported that DNA catalysts (deoxyribozymes) can hydrolyze DNA phosphodiester linkages, but DNA-catalyzed amide bond hydrolysis has been elusive. Here we used in vitro selection to identify DNA catalysts that hydrolyze ester linkages as well as DNA catalysts that hydrolyze aromatic amides, for which the leaving group is an aniline moiety. The aromatic amide-hydrolyzing deoxyribozymes were examined using linear free energy relationship analysis. The hydrolysis reaction is unaffec...

  4. Hydrolysis of Polysaccharides with 77% Sulfuric Acid for Quantitative Saccharification

    UÇAR, Güneş; Balaban, Mualla


    Classical standard hydrolysis of polysaccharides with 72% sulfuric acid was modified in 2 manners. In order to avoid treatment in an autoclave at 120 °C under pressure, wood or pulp material was first swollen in cold 77% acid followed by hydrolysis steps in diluted acid solutions. Further, the neutralization of the hydrolyzate with dilute barium hydroxide was carried out in heated mother liquor ensuring a crystalline precipitate of barium sulfate. Digestion enables the separation of clear ali...

  5. Optimization of enzymatic hydrolysis of fibre sludge from pulp mill

    Zinchenko, Ganna


    This thesis is a part of a project which aims at utilizing fibre sludge from pulp mill as a source of biofuel production. The study concentrates on optimizing one of the processing steps, enzymatic hydrolysis, in converting fibre sludge to bioalcohol. The aim of the thesis was to find optimum process parameters that enable maximum yield of glucose after performing the enzymatic hydrolysis. For this purpose, a series of experiments with changed process parameters was conducted. Also, enzym...

  6. The Mechanisms of Plant Cell Wall Deconstruction during Enzymatic Hydrolysis

    Thygesen, Lisbeth G; Thybring, Emil E.; Johansen, Katja S.; Claus Felby


    Mechanical agitation during enzymatic hydrolysis of insoluble plant biomass at high dry matter contents is indispensable for the initial liquefaction step in biorefining. It is known that particle size reduction is an important part of liquefaction, but the mechanisms involved are poorly understood. Here we put forward a simple model based on mechanical principles capable of capturing the result of the interaction between mechanical forces and cell wall weakening via hydrolysis of glucosidic ...

  7. Alkali pretreated of wheat straw and its enzymatic hydrolysis

    Lirong Han; Juntao Feng; Shuangxi Zhang; Zhiqing Ma; Yonghong Wang; Xing Zhang


    The efficiency of enzymatic hydrolysis of cellulose can be improved by various pretreatments of the substrate. In order to increase the efficiency of enzymatic saccharification of the wheat straw, we determined the effect of different pretreatments on the physical structure, chemical components and enzymatic saccharification of wheat straw. Our results showed that combination of grinding and sodium hydroxide (NaOH) treatment had high effect on the enzymatic hydrolysis of wheat straws. The opt...

  8. Direct injection of superheated steam for continuous hydrolysis reaction

    Wang, Weicheng


    The primary intent for previous continuous hydrolysis studies was to minimize the reaction temperature and reaction time. In this work, hydrolysis is the first step of a proprietary chemical process to convert lipids to sustainable, drop-in replacements for petroleum based fuels. To improve the economics of the process, attention is now focused on optimizing the energy efficiency of the process, maximizing the reaction rate, and improving the recovery of the glycerol by-product. A laboratory-scale reactor system has been designed and built with this goal in mind.Sweet water (water with glycerol from the hydrolysis reaction) is routed to a distillation column and heated above the boiling point of water at the reaction pressure. The steam pressure allows the steam to return to the reactor without pumping. Direct injection of steam into the hydrolysis reactor is shown to provide favorable equilibrium conditions resulting in a high quality of FFA product and rapid reaction rate, even without preheating the inlet water and oil and with lower reactor temperatures and lower fresh water demand. The high enthalpy of the steam provides energy for the hydrolysis reaction. Steam injection offers enhanced conditions for continuous hydrolysis of triglycerides to high-purity streams of FFA and glycerol. © 2012 Elsevier B.V.

  9. Site- and species-specific hydrolysis rates of heroin.

    Szöcs, Levente; Orgován, Gábor; Tóth, Gergő; Kraszni, Márta; Gergó, Lajos; Hosztafi, Sándor; Noszál, Béla


    The hydroxide-catalyzed non-enzymatic, simultaneous and consecutive hydrolyses of diacetylmorphine (DAM, heroin) are quantified in terms of 10 site- and species-specific rate constants in connection with also 10 site- and species-specific acid-base equilibrium constants, comprising all the 12 coexisting species in solution. This characterization involves the major and minor decomposition pathways via 6-acetylmorphine and 3-acetylmorphine, respectively, and morphine, the final product. Hydrolysis has been found to be 18-120 times faster at site 3 than at site 6, depending on the status of the amino group and the rest of the molecule. Nitrogen protonation accelerates the hydrolysis 5-6 times at site 3 and slightly less at site 6. Hydrolysis rate constants are interpreted in terms of intramolecular inductive effects and the concomitant local electron densities. Hydrolysis fraction, a new physico-chemical parameter is introduced and determined to quantify the contribution of the individual microspecies to the overall hydrolysis. Hydrolysis fractions are depicted as a function of pH. PMID:27130543

  10. Development of complete hydrolysis of pectins from apple pomace.

    Wikiera, Agnieszka; Mika, Magdalena; Starzyńska-Janiszewska, Anna; Stodolak, Bożena


    Enzymatically extracted pectins have a more complex structure than those obtained by conventional methods. As a result, they are less susceptible to hydrolysis, which makes the precise determination of their composition difficult. The aim of the study was to develop a method of complete hydrolysis of enzymatically extracted apple pectins. Substrates were pectins isolated from apple pomace by the use of xylanase and multicatalytic preparation Celluclast and apple pomace. Hydrolysis was performed by a chemical method with 2M TFA at 100 °C and 120 °C and a combined acidic/enzymatic method. After hydrolysis, the contents of galacturonic acid and neutral sugars were measured by HPLC. Complete hydrolysis of polygalacturonic acid occurred after 2.5h incubation with 2M TFA at 120 °C. The efficient hydrolysis of neutral sugars in pectins was performed with 2M TFA at 100 °C for 2.5h. Monomers most susceptible to concentrated acid were rhamnose, mannose and arabinose. PMID:25442606

  11. Nicosulfuron: alcoholysis, chemical hydrolysis, and degradation on various minerals.

    Sabadie, Jean


    Alcoholysis (methanol or ethanol) and hydrolysis (pH ranging from 4 to 11) of the herbicide nicosulfuron at 30 degrees C principally involves the breakdown of the urea part of the molecule. A high yield of the corresponding carbamate was obtained along with aminopyrimidine during alcoholysis. Hydrolysis led to both aminopyrimidine and pyridylsulfonamide. The latter compound may be easily cyclized (pH > or = 7). First-order kinetics describe the rates of alcoholysis and hydrolysis well. The rate constants (0.44 days(-1) for methanolysis) decreased from 0.50 to 0.002 days(-1) as pH increased from 4 to 8, then remained stable under alkaline conditions. In acidic or neutral solution, the hydrolysis path appeared prevalent (> or =70%), whereas in an alkaline medium it decreased when pH increased. The chemical degradation of nicosulfuron on various dry minerals (calcium bentonite, kaolinite, silica gel, H(+) bentonite, montmorillonite K10, and alumina) was investigated at 30 degrees C. The best conditions for the degradation are obtained on acidic minerals after herbicide deposition using the liquid method. Under these conditions an acceptable correlation with pseudo-first-order kinetics was observed, and the major degradation path is similar to that proposed for chemical hydrolysis. Conversely, alumina seemed to favor other unknown degradation processes. The hydrolysis paths of nicosulfuron and rimsulfuron appeared to be different. PMID:11804524

  12. Milk Lactose Hydrolysis In A Batch Reactor: Optimisation Of Process Parameters, Kinetics Of Hydrolysis And Enzyme Inactivation

    Sener, N.; Kilic-Apar, D.; DEMIRHAN, E.; Ozbek, B.


    The present investigation describes the effects of the process quantities on enzymatic hydrolysis of milk lactose and enzyme stability. The lactose hydrolysis reactions were carried out in 250 mL of milk by using a commercial β-galactosidase produced from Kluyveromyces marxianus lactis. The residual lactose mass concentration (g L-1) and residual enzyme activity (%) against time were investigated vs. process variables such as temperature, impeller speed and enzyme concentration. Optimum condi...

  13. Impacts of microalgae pre-treatments for improved anaerobic digestion: Thermal treatment, thermal hydrolysis, ultrasound and enzymatic hydrolysis

    Ometto, Francesco; Quiroga, Gerardo; Psenǐckǎ, Pavel; Whitton, Rachel; Jefferson, Bruce; Villa, Raffaella


    Anaerobic digestion (AD) of microalgae is primarily inhibited by the chemical composition of their cell walls containing biopolymers able to resist bacterial degradation. Adoption of pre-treatments such as thermal, thermal hydrolysis, ultrasound and enzymatic hydrolysis have the potential to remove these inhibitory compounds and enhance biogas yields by degrading the cell wall, and releasing the intracellular algogenic organic matter (AOM). This work investigated the effect of four pre-treatm...

  14. Stochastic molecular model of enzymatic hydrolysis of cellulose for ethanol production

    Kumar, Deepak; Murthy, Ganti S.


    Background During cellulosic ethanol production, cellulose hydrolysis is achieved by synergistic action of cellulase enzyme complex consisting of multiple enzymes with different mode of actions. Enzymatic hydrolysis of cellulose is one of the bottlenecks in the commercialization of the process due to low hydrolysis rates and high cost of enzymes. A robust hydrolysis model that can predict hydrolysis profile under various scenarios can act as an important forecasting tool to improve the hydrol...

  15. Enteral Tube Feeding Nutritional Protein Hydrolysate Production Under Different Factors By Enzymatic Hydrolysis

    Nguyen ThiQuynhHoa; Nguyen Ngoc Phuong Diem; Nguyen Phuoc Minh; Dong ThiAnh Dao


    Abstract Hydrolysis of proteins involves the cleavage of peptide bonds to give peptides of varying sizes and amino acid composition. There are a number of types of hydrolysis enzymatic acid or alkali hydrolysis. Chemical hydrolysis is difficult to control and reduces the nutritional quality of products destroying L-form amino acids and producing toxic substances such as lysino-alanine. Enzymatic hydrolysis works without destructing amino acids and by avoiding the extreme temperatures and pH l...

  16. Liquid chromatography-tandem mass spectrometry for the determination of sphingomyelin species from calf brain, ox liver, egg yolk, and krill oil.

    Zhou, Li; Zhao, Minjie; Ennahar, Saïd; Bindler, Françoise; Marchioni, Eric


    In this study, molecular species of sphingomyelin (SM) in egg yolk, calf brain, ox liver, and krill oil were investigated. Classes of phospholipids (PLs) were purified, identified, and quantified by normal phase semipreparative high-performance liquid chromatography (HPLC) combined with evaporative light scattering detectors (ELSD). For SM molecular species identification, pure SM collected through a flow splitter was loaded to HPLC-electrospray ionization-tandem mass spectrometry (LC-ESI-MS(2)), with 100% methanol containing 5 mM ammonium formate as mobile phase. In addition to classes of PLs, the used approach allowed the determination of profiles of SM species in egg yolk, ox liver, and calf brain, whereas krill oil turned out not to contain any SM. It also allowed the separation and identification of SM subclasses, as well as tentative identification of species with the same molecular mass, including isomers. The results showed that egg yolk contained the highest proportion of (d18:1-16:0)SM (94.1%). The major SM molecular species in ox liver were (d18:1-16:0)SM (25.5%), (d18:1-23:0)SM (19.7%), (d18:1-24:0)SM (13.2%), and (d18:1-22:0)SM (12.5%). Calf brain SM was rich in species such as (d18:1-18:0)SM (40.7%), (d18:1-24:1)SM (17.1%), and (d18:1-20:0)SM (10.8%). PMID:22148474

  17. Enzymatic Hydrolysis of Hydrotropic Pulps at Different Substrate Loadings.

    Denisova, Marina N; Makarova, Ekaterina I; Pavlov, Igor N; Budaeva, Vera V; Sakovich, Gennady V


    Enzymatic hydrolysis of cellulosic raw materials to produce nutrient broths for microbiological synthesis of ethanol and other valuable products is an important field of modern biotechnology. Biotechnological processing implies the selection of an effective pretreatment technique for raw materials. In this study, the hydrotropic treatment increased the reactivity of the obtained substrates toward enzymatic hydrolysis by 7.1 times for Miscanthus and by 7.3 times for oat hulls. The hydrotropic pulp from oat hulls was more reactive toward enzymatic hydrolysis compared to that from Miscanthus, despite that the substrates had similar compositions. As the initial substrate loadings were raised during enzymatic hydrolysis of the hydrotropic Miscanthus and oat hull pulps, the concentration of reducing sugars increased by 34 g/dm(3) and the yield of reducing sugars decreased by 31 %. The findings allow us to predict the efficiency of enzymatic hydrolysis of hydrotropic pulps from Miscanthus and oat hulls when scaling up the process by volume. PMID:26634840

  18. Enzymatic hydrolysis of fructans in the tequila production process.

    Avila-Fernández, Angela; Rendón-Poujol, Xóchitl; Olvera, Clarita; González, Fernando; Capella, Santiago; Peña-Alvarez, Araceli; López-Munguía, Agustín


    In contrast to the hydrolysis of reserve carbohydrates in most plant-derived alcoholic beverage processes carried out with enzymes, agave fructans in tequila production have traditionally been transformed to fermentable sugars through acid thermal hydrolysis. Experiments at the bench scale demonstrated that the extraction and hydrolysis of agave fructans can be carried out continuously using commercial inulinases in a countercurrent extraction process with shredded agave fibers. Difficulties in the temperature control of large extraction diffusers did not allow the scaling up of this procedure. Nevertheless, batch enzymatic hydrolysis of agave extracts obtained in diffusers operating at 60 and 90 degrees C was studied at the laboratory and industrial levels. The effects of the enzymatic process on some tequila congeners were studied, demonstrating that although a short thermal treatment is essential for the development of tequila's organoleptic characteristics, the fructan hydrolysis can be performed with enzymes without major modifications in the flavor or aroma, as determined by a plant sensory panel and corroborated by the analysis of tequila congeners. PMID:19473003

  19. Effects of structural features of cotton cellulose on enzymatic hydrolysis

    Textile cotton wastes were treated with γ rays and 18% NaOH and 70% ZnCl2 solutions and were subjected to enzymatic hydrolysis. The untreated and treated samples were characterized both before and after hydrolysis by means of parameters concerning molecular structure (degree of polymerization), supermolecular structure (x-ray diffraction), accessibility, and reactivity (moisture regain, enzyme adsorption, and solubility in FeTNa). These parameters were correlated to kinetic parameters of the hydrolysis reaction. The V/sub max/ and K/sub m/ values were evaluated from Lineweaver-Burk plots at different temperatures. The V/sub max//K/sub m/ ratio, analogous to the specificity constant, proved to be less sensitive to experimental errors and more suitable for a comparison of the kinetic behavior of the samples. The modifications of both supermolecular structure and morphology of cellulose were of primary importance to attain high yields and rates of hydrolysis. Furthermore, the structural and morphologic parameters chosen to characterize the samples can be correlated to the kinetic parameters of enzymatic hydrolysis, in particular to K/sub m/ values

  20. Linking hydrolysis performance to Trichoderma reesei cellulolytic enzyme profile.

    Lehmann, Linda; Rønnest, Nanna P; Jørgensen, Christian I; Olsson, Lisbeth; Stocks, Stuart M; Jørgensen, Henrik S; Hobley, Timothy


    Trichoderma reesei expresses a large number of enzymes involved in lignocellulose hydrolysis and the mechanism of how these enzymes work together is too complex to study by traditional methods, for example, by spiking with single enzymes and monitoring hydrolysis performance. In this study, a multivariate approach, partial least squares regression, was used to see whether it could help explain the correlation between enzyme profile and hydrolysis performance. Diverse enzyme mixtures were produced by T. reesei Rut-C30 by exploiting various fermentation conditions and used for hydrolysis of washed pretreated corn stover as a measure of enzyme performance. In addition, the enzyme mixtures were analyzed by liquid chromatography-tandem mass spectrometry to identify and quantify the different proteins. A multivariate model was applied for the prediction of enzyme performance based on the combination of different proteins present in an enzyme mixture. The multivariate model was used for identification of candidate proteins that are correlated to enzyme performance on pretreated corn stover. A very large variation in hydrolysis performance was observed and this was clearly caused by the difference in fermentation conditions. Besides β-glucosidase, the multivariate model identified several xylanases, Cip1 and Cip2, as relevant proteins to study further. Biotechnol. Bioeng. 2016;113: 1001-1010. © 2015 Wiley Periodicals, Inc. PMID:26524197

  1. Prediction of fetal lung maturity using the lecithin/sphingomyelin (L/S) ratio analysis with a simplified sample preparation, using a commercial microtip-column combined with mass spectrometric analysis.

    Kwak, Ho-Seok; Chung, Hee-Jung; Choi, Young Sik; Min, Won-Ki; Jung, So Young


    Fetal lung maturity is estimated using the lecithin/sphingomyelin ratio (L/S ratio) in amniotic fluid and it is commonly measured with thin-layer chromatography (TLC). The TLC method is time consuming and technically difficult; however, it is widely used because there is no alternative. We evaluated a novel method for measuring the L/S ratio, which involves a tip-column with a cation-exchange resin and mass spectrometry. Phospholipids in the amniotic fluid were extracted using methanol and chloroform. Choline-containing phospholipids such as lecithin and sphingomyelin were purified by passing them through the tip-column. LC-MS/MS and MALDI-TOF were used to directly analyze the purified samples. The L/S ratio by mass spectrometry was calculated from the sum peak intensity of the six lecithin, and that of sphingomyelin 34:1. In 20 samples, the L/S ratio determined with TLC was significantly correlated with that obtained by LC-MS/MS and MALDI-TOF. There was a 100% concordance between the L/S ratio by TLC and that by LC-MS/MS (kappa value=1.0). The concordance between the L/S ratio by TLC and that by MALDI-TOF was also 100% (kappa value=1.0). Our method provides a faster, simpler, and more reliable assessment of fetal lung maturity. The L/S ratio measured by LC-MS/MS and MALDI-TOF offers a compelling alternative method to traditional TLC. PMID:26000861

  2. Granular starch hydrolysis for fuel ethanol production

    Wang, Ping

    addition were evaluated in the dry grind process using GSHE (GSH process). Addition of proteases resulted in higher ethanol concentrations (15.2 to 18.0% v/v) and lower (DDGS) yields (32.9 to 45.8% db) compared to the control (no protease addition). As level of proteases and GSHE increased, ethanol concentrations increased and DDGS yields decreased. Proteases addition reduced required GSHE dose. Ethanol concentrations with protease addition alone were higher than with urea or with addition of both protease and urea. Corn endosperm consists of soft and hard endosperm. More exposed starch granules and rough surfaces produced from soft endosperm compared to hard endosperm will create more surface area which will benefit the solid phase hydrolysis as used in GSH process. In this study, the effects of protease, urea, endosperm hardness and GSHE levels on the GSH process were evaluated. Soft and hard endosperm materials were obtained by grinding and sifting flaking grits from dry milling pilot plant. Soft endosperm resulted in higher ethanol concentrations (at 72 hr) compared to ground corn or hard endosperm. Addition of urea increased ethanol concentrations (at 72 hr) for soft and hard endosperm. The effect of protease addition on increasing ethanol concentrations and fermentation rates was more predominant for soft endosperm, less for hard endosperm and least for ground corn. The GSH process with protease resulted in higher ethanol concentration than that with urea. For fermentation of soft endosperm, GSHE dose can be reduced. Ground corn fermented faster at the beginning than hard and soft endosperm due to the presence of inherent nutrients which enhanced yeast growth.

  3. Base hydrolysis and hydrothermal processing of PBX-9404

    Base hydrolysis in combination with hydrothermal processing has been proposed as an environmentally acceptable alternative to open burning/open detonation for degradation and destruction of high explosives. In this report, the authors examine gaseous and aqueous products of base hydrolysis of the HMX-based plastic bonded explosive, PBX-9404. They also examined products from the subsequent hydrothermal treatment of the base hydrolysate. The gases produced from hydrolysis of PBX-9404 are ammonia, nitrous oxide, and nitrogen. Major aqueous products are sodium formate, acetate, nitrate, and nitrite, but not all carbon products have been identified. Hydrothermal processing of base hydrolysate destroyed up to 98% of the organic carbon in solution, and higher destruction efficiencies are possible. Major gas products detected from hydrothermal processing were nitrogen and nitrous oxide

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

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


    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. PMID:24755261

  5. Comparative hydrolysis and fermentation of sugarcane and agave bagasse.

    Hernández-Salas, J M; Villa-Ramírez, M S; Veloz-Rendón, J S; Rivera-Hernández, K N; González-César, R A; Plascencia-Espinosa, M A; Trejo-Estrada, S R


    Sugarcane and agave bagasse samples were hydrolyzed with either mineral acids (HCl), commercial glucanases or a combined treatment consisting of alkaline delignification followed by enzymatic hydrolysis. Acid hydrolysis of sugar cane bagasse yielded a higher level of reducing sugars (37.21% for depithed bagasse and 35.37% for pith bagasse), when compared to metzal or metzontete (agave pinecone and leaves, 5.02% and 9.91%, respectively). An optimized enzyme formulation was used to process sugar cane bagasse, which contained Celluclast, Novozyme and Viscozyme L. From alkaline-enzymatic hydrolysis of sugarcane bagasse samples, a reduced level of reducing sugar yield was obtained (11-20%) compared to agave bagasse (12-58%). Selected hydrolyzates were fermented with a non-recombinant strain of Saccharomyces cerevisiae. Maximum alcohol yield by fermentation (32.6%) was obtained from the hydrolyzate of sugarcane depithed bagasse. Hydrolyzed agave waste residues provide an increased glucose decreased xylose product useful for biotechnological conversion. PMID:19000863

  6. Determining the hydrolysis of cations: A short overview

    Full text: The hydrolysis of metal ions is the most fundamental aqueous chemistry. As soon as the metal is introduced to water, dissolution may take place and if the water is pure only hydrolysis reactions will take place. There are several methods used in the literature to determine the stability constants of these reactions, e.g. solvent extraction, potentiometric titrations, ion exchange and solubility measurements. Which one to select is not straight forward. All of them have pros and cons and different regions of applicability with respect to whether they are good for determining the initial hydrolysis or the later stages. Once the constants are determined it is important to assess the uncertainty in the determination. We point out tools to make this straight forward and traceable which is most important in scientific studies. (authors)

  7. Acid hydrolysis of Biomass lignocellulose Onopordum nervosum Boiss

    Hydrolysis of resistant cellulose of Onopordum nervosum Boiss (thistle) to reducing sugars in dilute sulfuric acid in glass ampoules and long residence times has been studied and kinetic parameters determined. The rate of hydrolysis is similar to that of the cellulose of Douglas fir, but comparatively the effect of the acid is more pronounced than temperature. From kinetic data it can be pre ducted the yield and since it can be obtained at least 45% of the potential glucose (48% as reducing sugars) at 190 degree centigree, 1,6% acid and 6,1 min. residence time, it indicates that the continuous acid hydrolysis of thistle may be a process of commercial interest. (Author) 18 refs

  8. The optimization of soybean oil hydrolysis reaction research

    The hydrolysis reaction of soybean oil was optimized. The concentration effect of ethanolic alkaline solution (KOH and NaOH) to the oil acidity was studied. The alkaline concentrations, reaction time and temperature factors was investigated during the optimization of the hydrolysis or saponification reaction. KOH solution of 1 M showed a good saponification activity which resulted oil acid value of 226.8 mg/ g compared to NaOH solution with acid value of 225.4 mg/ g for the same reaction. The optimum saponification reaction of soybean oil occurred at 60 degree Celsius in 30 minutes by using ethanolic KOH 1 M with acid value of 229.6 mg/ g. Composition of free fatty acid before and after hydrolysis were determined by using gas chromatography. (author)


    Jianxin Jiang


    Full Text Available The enzymatic saccharification of pretreated furfural residues with different lignin content was studied to verify the effect of lignin removal in the hydrolysis process. The results showed that the glucose yield was improved by increasing the lignin removal. A maximum glucose yield of 96.8% was obtained when the residue with a lignin removal of 51.4% was hydrolyzed for 108 h at an enzyme loading of 25 FPU/g cellulose. However, further lignin removal did not increase the hydrolysis. The effect of enzyme loading on the enzymatic hydrolysis was also explored in this work. It was concluded that a high glucose yield of 90% was achieved when the enzyme dosage was reduced from 25 to 15 FPU/g cellulose, which was cost-effective for the sugar and ethanol production. The structures of raw material and delignified samples were further characterized by XRD and scanning electron microscopy (SEM.

  10. Complex enzyme hydrolysis releases antioxidative phenolics from rice bran.

    Liu, Lei; Wen, Wei; Zhang, Ruifen; Wei, Zhencheng; Deng, Yuanyuan; Xiao, Juan; Zhang, Mingwei


    In this study, phenolic profiles and antioxidant activity of rice bran were analyzed following successive treatment by gelatinization, liquefaction and complex enzyme hydrolysis. Compared with gelatinization alone, liquefaction slightly increased the total amount of phenolics and antioxidant activity as measured by ferric reducing antioxidant power (FRAP) and oxygen radical absorbance capacity (ORAC) assays. Complex enzyme hydrolysis significantly increased the total phenolics, flavonoids, FRAP and ORAC by 46.24%, 79.13%, 159.14% and 41.98%, respectively, compared to gelatinization alone. Furthermore, ten individual phenolics present in free or soluble conjugate forms were also analyzed following enzymatic processing. Ferulic acid experienced the largest release, followed by protocatechuic acid and then quercetin. Interestingly, a major proportion of phenolics existed as soluble conjugates, rather than free form. Overall, complex enzyme hydrolysis releases phenolics, thus increasing the antioxidant activity of rice bran extract. This study provides useful information for processing rice bran into functional beverage rich in phenolics. PMID:27507440

  11. Hydrolysis of starch by sorghum malt for maltodextrin production [abstract

    Thonart, P.


    Full Text Available Maltodextrin is a mixture of saccharides with a molecular weight between polysaccharides and oligosaccharides with DE lower than 20. Maltodextrin is more soluble in water than native starches, also is cheaper in comparison with other major edible hydrocolloids. Maltodextrin is obtained by moderate enzymatic or acidic hydrolysis of starch. The hydrolysis of starch, catalyzed by amylases, is the most important commercial enzyme process. The hydrolyzed products are widely applied in food, paper and textile industries. Because of the increasing demand for these enzymes in various industries, there is enormous interest in developing enzymes with better properties, such as raw starch-degrading amylases suitable for industrial applications and their cost-effective production techniques. Sorghum (Sorghum bicolor is a widely grown crop in Africa. Obtaining enzymes from sorghum requires a transformation. The objective of this study was application of sorghum amylase for maltodextrin. Sorghum seeds were supplied by the ISRA (Bambey, Senegal. Seeds were germinated in the laboratory at 30°C for 72 h and the sorghum malt was dried at 40°C for 48 h. Corn starch (from Roquette, France hydrolysis was assayed in a bioreactor of 2 l at a temperature of 65°C gently stirred. Raw starch was slurried in water (30% w/v and sorghum malt was introduced, chloride calcium was added, pH was adjusted to 6. Maltodextrin was characterized in term of the dextrose equivalent (DE during the hydrolysis. The yield of hydrolysis was evaluated by soluble solids (° BRIX at different hydrolysis time. The glucose concentration released was measured by DNS method (Miller method.

  12. Visualizing phosphodiester-bond hydrolysis by an endonuclease

    Molina, Rafael; Stella, Stefano; Redondo, Pilar;


    The enzymatic hydrolysis of DNA phosphodiester bonds has been widely studied, but the chemical reaction has not yet been observed. Here we follow the generation of a DNA double-strand break (DSB) by the Desulfurococcus mobilis homing endonuclease I-DmoI, trapping sequential stages of a two...... third metal ion has a crucial role, triggering the consecutive hydrolysis of the targeted phosphodiester bonds in the DNA strands and leaving its position once the DSB is generated. The multiple structures show the orchestrated conformational changes in the protein residues, nucleotides and metals...

  13. The hydrolysis of aluminium, a mass spectrometric study

    Sarpola, A.


    Abstract This thesis is focused on the hydrolysis of aluminium, the polymerisation of the hydrolysis products, and how these can be monitored by mass spectrometric methods. The main aim of this research is to figure out how the aqueous speciation of aluminium changes as a function of pH (3.2–10), concentration (1–100 mM), reaction time (1s–14d), and counter anion (Cl-, SO42-, HCOO-). The method used was electrospray mass spectrometry. The results showed more variable speciation than those ...

  14. Combined wet oxidation and alkaline hydrolysis of polyvinylchloride

    Sørensen, E.; Bjerre, A.B.


    In view of the widespread aversion to burning polyvinylchloride (PVC) together with municipal waste, we have attempted an alternative approach to its decomposition. This paper describes a combined wet oxidation/alkaline hydrolysis yielding water soluble, biodegradable products. Experiments were...... carried out at temperatures from 180-260 degree C and reaction times of 8-24 min. The chloride liberated provides information on the rate constants. Considering the measured Cl- and Chemical Oxygen Demand (COD) values, we find hydrolysis and oxidation processes to be interdependent. The main products are...

  15. Enzymatic hydrolysis of potato starch and ethanol production

    Lazić Miodrag L.; Rašković Suzana; Stanković Mihajlo Z.; Veljković Vlada B.


    The hydrolysis of potato starch using one (Termamyl or Fungamyl) and two combined (Termamyl and Supersan) commercial enzyme preparations and ethanol production from the hydrolysates obtained using the yeast Saccharomyces cerevisiae were studied. Potato tubers were previously prepared as mash or flour. The study dealt with the effects of the hydromodulus (1:1 and 1:0.5), particle size (0.1, 0.2 and 0.4 mm) as well as the type and concentration of enzyme on the enzymatic hydrolysis of potato st...

  16. Kinetics and mechanism of hydrolysis of scandium sulfate

    The Sc2(SO4)3-H2SO4-H2O system is studied through the methods of pH-potentiometry, conductometry and turbidimetry at 298 and 318 K and ion force 0.01, 0.1 and 1.0. The hydrolysis mechanism including the processes in the system homogenous and heterogeneous constituents. The hydrolysis rates of scandium salts and their dependences on OH-ions concentration, solution ions force and temperature are found; the constants of the processes rate with participation of OH- and SO42- ions and constants of the solid phase formation rate are calculated

  17. Benzene/nitrous oxide flammability in the precipitate hydrolysis process

    Jacobs, R A [Du Pont de Nemours (E.I.) and Co., Aiken, SC (USA). Savannah River Lab.


    The HAN (hydroxylamine nitrate) process for destruction of nitrite in precipitate hydrolysis produces nitrous oxide (N2O) gas as one of the products. N2O can form flammable mixtures with benzene which is also present due to radiolysis and hydrolysis of tetraphenylborate. Extensive flame modeling and explosion testing was undertaken to define the minimum oxidant for combustion of N2O/benzene using both nitrogen and carbon dioxide as diluents. The attached memorandum interprets and documents the results of the studies.

  18. The Mechanisms of Plant Cell Wall Deconstruction during Enzymatic Hydrolysis

    Thygesen, Lisbeth Garbrecht; E. Thybring, Emil; Johansen, Katja Salomon;


    Mechanical agitation during enzymatic hydrolysis of insoluble plant biomass at high dry matter contents is indispensable for the initial liquefaction step in biorefining. It is known that particle size reduction is an important part of liquefaction, but the mechanisms involved are poorly understood....... Here we put forward a simple model based on mechanical principles capable of capturing the result of the interaction between mechanical forces and cell wall weakening via hydrolysis of glucosidic bonds. This study illustrates that basic material science insights are relevant also within biochemistry...

  19. Enzymatic hydrolysis of fish frames using pilot plant scale systems

    Himonides, Aristotelis T.; Taylor, Anthony K. D.; Morris, Anne J.


    Papain was used to hydrolyse fish frames under controlled conditions at a batch-pilot plant scale-process, for the pro-duction of fish protein hydrolysates (FPH). Mass balance calculations were carried out so that the rate of hydrolysis, rate of protein solubilisation and yields could be estimated. Almost complete hydrolysis could be achieved in 1 hour, at 40°C, with no pH adjustment, at 0.5% (5 g·kg−1) enzyme to substrate ratio (E/S, were S is Kjeldahl protein) using whole fish frames (inclu...

  20. Reaction pathways and free energy profiles for spontaneous hydrolysis of urea and tetramethylurea: Unexpected substituent effects

    Yao, Min; Tu, Wenlong; Chen, Xi; Zhan, Chang-Guo


    It has been difficult to directly measure the spontaneous hydrolysis rate of urea and, thus, 1,1,3,3-tetramethylurea (Me4U) was used as a model to determine the “experimental” rate constant for urea hydrolysis. The use of Me4U was based on an assumption that the rate of urea hydrolysis should be 2.8 times that of Me4U hydrolysis because the rate of acetamide hydrolysis is 2.8 times that of N,N-dimethyl-acetamide hydrolysis. The present first-principles electronic-structure calculations on the...

  1. Intramolecular Amide Hydrolysis in N-Methylmaleamic Acid Revisited


    The intramolecular amide hydrolysis of N-methylmaleamic acid have been revisited by use of density functional theory and inclusion of solvent effects. The results indicate that concerted reaction mechanism is favored over stepwise reaction mechanism. This is in agreement with the previous theoretical study. Sovlent effects have significant influence on the reaction barrier.

  2. Theoretical Aspects of Hydrolysis of Peptide Bonds by Zinc Metalloenzymes

    Navrátil, Václav; Klusák, Vojtěch; Rulíšek, Lubomír


    Roč. 19, č. 49 (2013), s. 16634-16645. ISSN 0947-6539 Institutional support: RVO:61388963 Keywords : ab initio calculations * hydrolysis * metalloenzymes * peptides * transition states Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 5.696, year: 2013

  3. Hydrolysis of aluminum nitride powders in moist air

    High thermal conductivity is required for successful application of aluminum nitride (AlN) as a substrate material in electronic devices. AlN powders of low oxygen content are needed since oxygen contamination greatly reduces the thermal conductivity of AlN ceramics. High-purity AlN powders are commercially available, but can be contaminated by oxygen when contacting water/oxygen in powder processing after manufacturing. The present study investigates the hydrolysis properties of AlN powders in moist air at room temperature, so as to understand the degradation phenomena during powder handling in the normal atmospheric environment. The powders investigated were produced via three major commercial processes, namely, chemical vapor deposition from triethyl aluminum, carbothermal reduction and nitridation of alumina, and direct nitridation of aluminum. At the beginning of hydrolysis, an induction period is observed for each powder, which is attributed to slow hydrolysis of the surface oxide/oxyhydroxide layer. The length of this period is thus dependent on the composition and thickness of the surface layer, which is in turn affected by the manufacturing method. The AlN powder produced by the carbothermal process shows the longest induction period. The hydrolysis reaction produces initially amorphous AlOOH, which is further hydrolyzed to mixtures of bayerite, nordstrandite, and gibbsite, forming agglomerates around the unreacted AlN. Copyright (2003) AD-TECH - International Foundation for the Advancement of Technology Ltd

  4. Penicillin Hydrolysis: A Kinetic Study of a Multistep, Multiproduct Reaction.

    McCarrick, Thomas A.; McLafferty, Fred W.


    Background, procedures used, and typical results are provided for an experiment in which students carry out the necessary measurements on the acid-catalysis of penicillin in two hours. By applying kinetic theory to the data obtained, the reaction pathways for the hydrolysis of potassium benzyl penicillin are elucidated. (JN)

  5. Efficiency of hydrolysis of whey protein serine proteases

    Г.П. Петюх; Романова, Н.А.; К.М. Гаркава


     Research of relative enzymatic whey protein isolate (WPI) hydrolysis efficiency was conducted. Seven enzymes in three concentrations by pH 7.0 t=50°C were used. The most efficient showed to be Protease R in concentration 0,0002 g/ml.

  6. Starch hydrolysis under low water conditions: a conceptual process design

    Veen, van der M.E.; Veelaert, S.; Goot, van der A.J.; Boom, R.M.


    A process concept is presented for the hydrolysis of starch to glucose in highly concentrated systems. Depending on the moisture content, the process consists of two or three stages. The two-stage process comprises combined thermal and enzymatic liquefaction, followed by enzymatic saccharification.

  7. Influence of water availability on the enzymatic hydrolysis of proteins

    Butré, C.I.; Wierenga, P.A.; Gruppen, H.


    The overall rate of enzymatic protein hydrolysis decreases with increasing protein concentration (0.1–30% (w/v)) at constant enzyme/substrate ratio. To understand the role of water, the amount of available water was expressed as the ratio between free and bound water and experimentally determined fr

  8. Wet explosion pretreatment of sugarcane bagasse for enhanced enzymatic hydrolysis

    Biswas, Rajib; Uellendahl, Hinrich; Ahring, Birgitte Kiær


    Wet explosion pretreatment of sugarcane bagasse was investigated in pilot-scale with the aim of obtaining the highest possible sugar yield after pretreatment. The temperatures used were 155, 170, 185 and 200 C with or without addition of oxygen (0.6 MPa pressure). Enzymatic hydrolysis of washed...

  9. Heteropoly acid catalyzed hydrolysis of glycogen to glucose

    Complete conversion of glycogen to glucose is achieved by using H3PW12O40·nH2O (HPW) and H4SiW12O40·nH2O (HSiW) as catalysts for the hydrolysis under optimized hydrothermal conditions (mass fraction of catalyst 2.4%, 373 K and 2 h reaction time). The reusability of the catalyst (HPW) was demonstrated. In addition to carrying out the glycogen hydrolysis in an autoclave, other novel methods such as microwave irradiation and sonication have also been investigated. At higher mass fraction of the heteropoly acids (10.5%), glycogen could be completely converted to glucose under microwave irradiation. Sonication of an aqueous solution of glycogen in the presence of HPW and HSiW also yielded glucose. Thus, heteropoly acids are efficient, environmentally friendly and reusable catalysts for the conversion of glycogen to glucose. - Highlights: • Hydrothermal, microwave and sonication based methods of hydrolysis. • Heteropoly acids are green catalysts for glycogen hydrolysis. • Glycogen from cyanobacteria is demonstrated as a potential feedstock for glucose

  10. Single Molecule Study of Cellulase Hydrolysis of Crystalline Cellulose

    Liu, Y.-S.; Luo, Y.; Baker, J. O.; Zeng, Y.; Himmel, M. E.; Smith, S.; Ding, S.-Y.


    This report seeks to elucidate the role of cellobiohydrolase-I (CBH I) in the hydrolysis of crystalline cellulose. A single-molecule approach uses various imaging techniques to investigate the surface structure of crystalline cellulose and changes made in the structure by CBH I.

  11. Catalytic hydrolysis for the degradation of organophosphorus pesticides in water

    The kinetic studies of catalytic hydrolysis revealed that the concentration of two kinds of organophosphorus pesticides (omethoate and methidathion) in solution apparently decays according to the second order reaction. It was found that the rate constant value was highest at strong acidic conditions and it continued to decrease as the pH of the solution was increased. At basic conditions the rate constant value decreased to minimum. Manganese dioxide under acidic conditions converted into Mn/sup 2+/ ions and then these ions in water form hexaaquomanganese (II) ion. This hexaaquomanganese (II ion then adsorbed itself on the S or O atom of the organophosphorus compound and thus weakens the bond between P-S. This reaction facilitated the attack of H/sub 2/O or OH/sup -/ ion and thus enhanced the efficiency of hydrolysis. It was studied that methidathion hydrolyzed more efficiently than omethoate The rate constants of catalytic hydrolysis were increased with increasing the amount of MnO/sub 2/. It was found that the pesticides had undergone adsorption on catalyst in the first few minutes and there was the rapid drop of total phosphorus concentration. The decrease of total phosphorus adsorption with increasing pH was also observed. After the addition of alkaline earth metal cations (Ca/sup 2+/ and Mg/sup 2+/) along with magnesium, the enhancement in the efficiency of hydrolysis at near neutral conditions occurred. (author)

  12. Hydrolysis of toxic natural glucosides catalyzed by cyclodextrin dicyanohydrins

    Bjerre, J.; Nielsen, Erik Holm Toustrup; Bols, M.


    The hydrolysis of toxic 7-hydroxycoumarin glucosides and other aryl and alkyl glucosides, catalyzed by modified a- and ß-cyclodextrin dicyanohydrins, was investigated using different UV, redox, or HPAEC detection assays. The catalyzed reactions all followed Michaelis-Menten kinetics, and an...

  13. Effects of hydrolysis and carbonization reactions on hydrochar production.

    Fakkaew, K; Koottatep, T; Polprasert, C


    Hydrothermal carbonization (HTC) is a thermal conversion process which converts wet biomass into hydrochar. In this study, a low-energy HTC process named "Two-stage HTC" comprising of hydrolysis and carbonization stages using faecal sludge as feedstock was developed and optimized. The experimental results indicated the optimum conditions of the two-stage HTC to be; hydrolysis temperature of 170 °C, hydrolysis reaction time of 155 min, carbonization temperature of 215 °C, and carbonization reaction time of 100 min. The hydrolysis reaction time and carbonization temperature had a statistically significant effect on energy content of the produced hydrochar. Energy input of the two-stage HTC was about 25% less than conventional HTC. Energy efficiency of the two-stage HTC for treating faecal sludge was higher than that of conventional HTC and other thermal conversion processes such as pyrolysis and gasification. The two-stage HTC could be considered as a potential technology for treating FS and producing hydrochar. PMID:26051497

  14. Chemical and enzymatic hydrolysis of anthraquinone glycosides from Madder roots

    Derksen, G.C.H.; Naayer, M.; Beek, T.A. van; Capelle, A.; Haaksman, I.K.; Doren, H.A. van; Groot, Æ. de


    For the production of a commercially useful dye extract from madder, the glycoside ruberythric acid has to be hydrolysed to the aglycone alizarin which is the main dye component. An intrinsic problem is the simultaneous hydrolysis of the glycoside lucidin pritneveroside to the unwanted mutagenic agl

  15. Chemical and enzymatic hydrolysis fo anthraquinone glycosides from madder roots

    Derksen, G.C.H.; Naayer, M.; Beek, van T.A.; Capelle, A.; Haaksman, I.K.; Doren, H.A.; Groot, de Æ.


    For the production of a commercially useful dye extract from madder, the glycoside ruberythric acid has to be hydrolysed to the aglycone alizarin which is the main dye component. An intrinsic problem is the simultaneous hydrolysis of the glycoside lucidin primeveroside to the unwanted mutagenic agly


    The acid-catalyzed hydrolysis of atrazine and related 2-chloro-s-triazines to the corresponding 2-hydroxy-s-triazines was investigated using the B3LYP hybrid density functional theory method. Gas-phase calculations were performed at the B3LYP/6-311++G(d,p)//B3LYP/6-31G* level of ...


    Farid Talebnia


    Full Text Available The effect of time, acid concentration, temperature and solid concentration on dilute-acid hydrolysis of orange peels was investigated. A central composite rotatable experimental design (CCRD was applied to study the individual effects of these hydrolysis factors and also their interdependence effects. The enzymatic hydrolysis of the peels by cellulase, β-glucosidase, and pectinase enzyme resulted in 72% dissolution of the peels, including 18.7% galacturonic acid and 53.3% of a total of glucose, fructose, galactose, and arabinose. Dilute-acid hydrolysis up to 210°C was not able to hydrolyze pectin to galacturonic acid. However, the sugar polymers were hydrolyzed at relatively low temperature. The optimum results were obtained at 116°C, 0.5% sulfuric acid concentration, 6% solid fraction, and 12.9 min retention time. Under these conditions, the total sugars obtained at 41.8 g/g dry peels and 2.6% of total hexose sugars were further degraded to hydroxymethylfurfural (HMF. No furfural was detected through these experiments from decomposition of pentoses.

  18. Small peptides hydrolysis in dry-cured meats.

    Mora, Leticia; Gallego, Marta; Escudero, Elizabeth; Reig, Milagro; Aristoy, M-Concepción; Toldrá, Fidel


    Large amounts of different peptides are naturally generated in dry-cured meats as a consequence of the intense proteolysis mechanisms which take place during their processing. In fact, meat proteins are extensively hydrolysed by muscle endo-peptidases (mainly calpains and cathepsins) followed by exo-peptidases (mainly, tri- and di-peptidyl peptidases, dipeptidases, aminopeptidases and carboxypeptidases). The result is a large amount of released free amino acids and a pool of numerous peptides with different sequences and lengths, some of them with interesting sequences for bioactivity. This manuscript is presenting the proteomic identification of small peptides resulting from the hydrolysis of four target proteins (glyceraldehyde-3-phosphate dehydrogenase, beta-enolase, myozenin-1 and troponin T) and discusses the enzymatic routes for their generation during the dry-curing process. The results indicate that the hydrolysis of peptides follows similar exo-peptidase mechanisms. In the case of dry-fermented sausages, most of the observed hydrolysis is the result of the combined action of muscle and microbial exo-peptidases except for the hydrolysis of di- and tri-peptides, mostly due to microbial di- and tri-peptidases, and the release of amino acids at the C-terminal that appears to be mostly due to muscle carboxypeptidases. PMID:25944374

  19. Lactam hydrolysis catalyzed by mononuclear metallo-ß-bactamases

    Olsen, Lars; Antony, J; Ryde, U;


    Two central steps in the hydrolysis of lactam antibiotics catalyzed by mononuclear metallo-beta-lactamases, formation of the tetrahedral intermediate and its breakdown by proton transfer, are studied for model systems using the density functional B3LYP method. Metallo-beta-lactamases have two met...

  20. Radioactive demonstration of the ''late wash'' Precipitate Hydrolysis Process

    This report presents results of the radioactive demonstration of the DWPF Precipitate Hydrolysis Process as it would occur in the ''late wash'' flowsheet in the absence of hydroxylamine nitrate. Radioactive precipitate containing Cs-137 from the April, 1983, in-tank precipitation demonstration in Tank 48 was used for these tests

  1. Influence of kaolinite on chiral hydrolysis of methyl dichlorprop enantiomers


    The effect of kaolinite on the enzymatic chiral hydrolysis of methyl dichlorprop enantiomers ((R,S)-methyl-2-(2,4-dichlorophenoxy) propanoic acid, 2,4-DPM) was investigated using chiral gas chromatography. Compared with the control without kaolinite, the enantiomeric ratio (ER) increased from 1.35 to 8.33 and the residual ratio of 2,4-DPM decreased from 60.89% to 41.55% in the presence of kaolinite. Kaolinite likely had emotion influence on lipase activity and its enantioselectivity.Moreover, the amount of kaolinite added was also found to be a sensitive factor affecting the enantioselective hydrolysis of 2,4-DPM. Fourier transform infrared (FTIR) spectroscopy studies of the interaction of lipase with kaolinite provided insight into the molecular structure of the complex and offered explanation of the effects of kaolinite on enzymatic hydrolysis of 2,4-DPM.Spectra showed that the effect of kaolinite on the hydrolysis of 2,4-DPM was affected by adsorption of lipase on kaolinite and changes of adsorbed lipase conformation, which led to the modified enantioselectivity.

  2. Enzymatic hydrolysis of protein:mechanism and kinetic model

    Qi Wei; He Zhimin


    The bioreaction mechanism and kinetic behavior of protein enzymatic hydrolysis for preparing active peptides were investigated to model and characterize the enzymatic hydrolysis curves.Taking into account single-substrate hydrolysis,enzyme inactivation and substrate or product inhibition,the reaction mechanism could be deduced from a series of experimental results carried out in a stirred tank reactor at different substrate concentrations,enzyme concentrations and temperatures based on M-M equation.An exponential equation dh/dt = aexp(-bh) was also established,where parameters a and b have different expressions according to different reaction mechanisms,and different values for different reaction systems.For BSA-trypsin model system,the regressive results agree with the experimental data,i.e.the average relative error was only 4.73%,and the reaction constants were determined as Km = 0.0748 g/L,Ks = 7.961 g/L,kd = 9.358/min,k2 =38.439/min,Ea= 64.826 kJ/mol,Ed= 80.031 kJ/mol in accordance with the proposed kinetic mode.The whole set of exponential kinetic equations can be used to model the bioreaction process of protein enzymatic hydrolysis,to calculate the thermodynamic and kinetic constants,and to optimize the operating parameters for bioreactor design.

  3. Optimization of enzymatic hydrolysis of cassava to obtain fermentable sugars

    Renata M. COLLARES; Luiza V. S. MIKLASEVICIUS; Mariana M. BASSACO; Nina P. G. SALAU; Marcio A. MAZUTTI; Dilson A. BISOGNIN; Lisiane M. TERRA


    This work evaluates the enzymatic hydrolysis of starch from cassava using pectinase,α-amylase,and amyloglucosidase.A central composite rotational design (CCRD) was carried out to evaluate the effects of amyloglucosidase,pectinase,reaction time,and solid to liquid ratio.All the experiments were carried out in a bioreactor with working volume of 2 L.Approximately 98% efficiency hydrolysis was obtained,resulting in a concentration of total reducing sugar released of 160 g/L.It was concluded that pectinase improved the hydrolysis of starch from cassava.Reaction time was found to be significant until 7 h of reaction.A solid to liquid ratio of 1.0 was considered suitable for hydrolysis of starch from cassava.Amyloglucosidase was a significant variable in the process:after its addition to the reaction media,a 30%-50% increase in the amount of total reducing sugar released was observed.At optimal conditions the maximum productivity obtained was 22.9 g/(L·h).

  4. Effect of particle size on enzymatic hydrolysis of pretreated Miscanthus

    Particle size reduction is a crucial factor in transportation logistics as well as cellulosic conversion. The effect of particle size on enzymatic hydrolysis of pretreated Miscanthus x giganteus was determined. Miscanthus was ground using a hammer mill equipped with screens having 0.08, 2.0 or 6.0...

  5. The Preparation and Enzymatic Hydrolysis of a Library of Esters

    Sanford, Elizabeth M.; Smith, Traci L.


    An investigative case study involving the preparation of a library of esters using Fischer esterification and alcoholysis of acid chlorides and their subsequent enzymatic hydrolysis by pig liver esterase and orange peel esterase is described. Students work collaboratively to prepare and characterize the library of esters and complete and evaluate…

  6. Lipase-catalyzed hydrolysis of methyl-3-phenylglycidate


    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.

  7. Bioabatement with xylanase supplementation to reduce enzymatic hydrolysis inhibitors

    Bioabatement, using the fungus Coniochaeta ligniaria NRRL30616 can effectively eliminate enzyme inhibitors from pretreated biomass hydrolysis. However, our recent research suggested that bioabatement had no beneficial effect on removing xylo-oligomers which were identified as strong inhibitors to ce...

  8. Mechanisms of lactone hydrolysis in neutral and alkaline conditions.

    Gómez-Bombarelli, Rafael; Calle, Emilio; Casado, Julio


    The neutral and base-catalyzed hydrolysis of nine carboxylic acid esters was studied using a hybrid supermolecule-PCM approach including six explicit water molecules. The molecules studied included two linear esters, four β-lactones, two γ-lactones, and one δ-lactone: ethyl acetate and methyl formate, β-propiolactone, β-butyrolactone, β-isovalerolactone, diketene (4-methyleneoxetan-2-one), γ-butyrolactone, 2(5H)-furanone, and δ-valerolactone. DFT and ab initio methods were used to analyze the features of the various possible hydrolysis mechanisms. For all compounds, reasonable to very good qualitative and quantitative agreement with experimental work was found, and evidence is provided to support long-standing hypotheses regarding the role of solvent molecule as a base catalyst. In addition, novel evidence is presented for the existence of an elimination-addition mechanism in the basic hydrolysis of diketene. A parallel work addresses the acid-catalyzed hydrolysis of lactones. PMID:23758295

  9. Effect of lignin structure on enzymatic hydrolysis of plant residues

    Sipponen, Mika


    Biochemical conversion of lignocellulose into high value and energy-intensive products necessitates pretreatments that enhance enzymatic hydrolysis of lignocellulosic carbohydrates. This thesis investigated structural changes in lignin during various analytical and industrially relevant treatments of crop residues. The objective was to elucidate the effect of lignin structure on enzymatic digestibility of cellulose. Fractionation of lignin during sequential alkaline treatments of maize ste...

  10. Hydrolysis reactions of Am(III) and Am(V)

    Hydrolysis reactions of Am(III) have been investigated in non-complexing solution as well as saline solutions under CO2-free conditions. The solubility experiment in combination with radiometric pH-titration is carried out for the pH range from 6 to 13, at different ionic strengths. In non-complexing solution, the solubility product is found to be increasing along with an increase of the specific α-activity in a given experimental solution. In concentrated NaCl solutions (I ≥ 3M, pH>7) with high specific α-activities (>1 Ci/L), the α-radiolysis starts generating substantial amounts of oxidized chlorine species, e.g. ClO-, which results in the oxidation of Am(III) to Am(V). The hydrolysis reaction of Am(V) in 3M NaCl is also investigated by radiometric pH-titration. Solubility products (Ksp) of Am(OH)3(s) and AmO2(OH)(s) and hydrolysis constants of Am(III) and Am(V) are calculated from the solubility data. The speciation of different hydrolysis products, undertaken by laser-induced photacoustic spectroscopy (LPAS), has verified probable species assessed theoretically on the basis of solubility experiments. (orig.)

  11. Effect of nitrogen oxide pretreatments on enzymatic hydrolysis of cellulose

    Borrevik, R.K.; Wilke, C.R.; Brink, D.L.


    This work considers the effect of nitrogen oxide pretreatments on the subsequent enzymatic hydrolysis by Trichoderma viride cellulase of the cellulose occurring in wheat straw; Triticum Aestivum-L, em. Thell. In the pretreatment scheme the straw is first reacted with nitric oxide and air, and then extracted in aqueous solution. In this way, overall sugar yields increased from 17% for the case of no pretreatment to 70%. The glucose yield increased from 20 to 60%. The yield of glucose during enzymatic hydrolysis is dependent on the reaction time of the gas phase reaction. For a 24 hour reaction the yield is 60%, but drops to 45% for a reaction time of 2 hours. Xylose, a potentially valuable side product of the pretreatment, is obtained by dilute acid hydrolysis during the extraction stage in yields of 90 to 96%. In acidic media, the kinetics of both the rate of formation and destruction of xylose were found to follow the first-order rate laws reported in the literature. These were determined to be 4.5 (liter/gmole)(hr./sup -1/) and 0.03 hr./sup -1/, respectively. However, the rate of formation is much greater (20.4 (liter/gmole) (hr./sup -1/)) when the extraction liquor is recycled. The most likely explanation for this is that the increased total acidity of the recycled liquor compensates for diffusional limitations. A preliminary design and cost analysis of the pretreatment-hydrolysis scheme indicates that glucose can be produced at 10.86 cents per pound, exclusive of straw cost. The corresponding cost per pound of total sugars produced is 5.0 cents. Sensitivity analyses indicate that 42% of the pretreatment cost (excluding hydrolysis) can be attributed to nitric oxide production, and the high yield of sugar obtained is advantageous when considering the cost of straw.

  12. Hydrolysis of thorium(iv) at variable temperatures.

    Zanonato, P L; Di Bernardo, P; Zhang, Z; Gong, Y; Tian, G; Gibson, J K; Rao, L


    Hydrolysis of Th(iv) was studied in tetraethylammonium perchlorate (0.10 mol kg(-1)) at variable temperatures (283-358 K) by potentiometry and microcalorimetry. Three hydrolysis reactions, mTh(4+) + nH2O = Thm(OH)n((4m-n)+) + nH(+), in which (n,m) = (2,2), (8,4), and (15,6), were invoked to describe the potentiometric and calorimetric data for solutions with the [hydroxide]/[Th(iv)] ratio ≤ 2. At higher ratios, the formation of (16,5) cannot be excluded. The hydrolysis constants, *β2,2, *β8,4, and *β15,6, increased by 3, 7, and 11 orders of magnitude, respectively, as the temperature was increased from 283 to 358 K. The enhancement is mainly due to the significant increase of the degree of ionization of water as the temperature rises. All three hydrolysis reactions are endothermic at 298 K, with enthalpies of (118 ± 4) kJ mol(-1), (236 ± 7) kJ mol(-1), and (554 ± 4) kJ mol(-1) for ΔH2,2, ΔH8,4, and ΔH15,6 respectively. The hydrolysis constants at infinite dilution have been obtained with the specific ion interaction approach. The applicability of three approaches for estimating the equilibrium constants at different temperatures, including the constant enthalpy approach, the constant heat capacity approach and the DQUANT equation was evaluated with the data from this work. PMID:27460458

  13. Generation of group B soyasaponins I and III by hydrolysis.

    Zhang, Wei; Teng, Su Ping; Popovich, David G


    Soyasaponins are a group of oleanane triterpenoids found in soy and other legumes that have been associated with some of the benefits achieved by consuming plant-based diets. However, these groups of compounds are diverse and structurally complicated to chemically characterize, separate from the isoflavones, and isolate in sufficient quantities for bioactive testing. Therefore, the aim of this study was to maximize the extraction of soyasaponins from soy flour, remove isoflavones, separate group B soyasaponins from group A, and produce an extract that contained a majority of non-DDMP (2,3-dihydro-2,5-dihydroxy-6-methyl-4H-pyran-4-one)-conjugated group B soyasaponins I and III. Room temperature extraction in methanol for 24 or 48 h resulted in the maximum recovery of soyasaponins, and Soxhlet extraction resulted in the least. A solid-phase extraction using methanol (45%) was found to virtually eliminate the interfering isoflavones as compared to butanol-water liquid-liquid extraction and ammonium sulfate precipitation, while maximizing saponin recovery. Alkaline hydrolysis in anhydrous methanol produced the maximum amount of soyasaponins I and III as compared to aqueous methanol and acid hydrolysis in both aqueous and anhydrous methanol. The soyasaponin I amount was increased by 175%, and soyasaponin III was increased by 211% after alkaline hydrolysis. Furthermore, after alkaline hydrolysis, a majority of DDMP-conjugated group B soyasaponins such as betag, betaa, gammag, and gammaa transformed into the non-DDMP-conjugated soyasaponins I and III without affecting the glycosidic bond at position C-3 of the ring structure. Therefore, we have developed a method that maximizes the recovery of DDMP-conjugated saponins and uses alkaline hydrolysis to produce an extract containing mainly soyasaponins I and III. PMID:19338335

  14. Bioethanol production: Pretreatment and enzymatic hydrolysis of softwood

    Tengborg, Charlotte


    The enzymatic hydrolysis process can be used to produce bioethanol from softwood, which are the dominating raw material in the Northern hemisphere. This thesis deals with the development of the process focusing on the pretreatment and the enzymatic hydrolysis stages. The influence of pretreatment conditions on sugar yield, and the effect of inhibitors on the ethanol yield, were investigated for spruce and pine. The maximum yields of hemicellulose sugars and glucose were obtained under different pretreatment conditions. This indicates that two-stage pretreatment may be preferable. The added catalysts, H{sub 2}SO{sub 4} and SO{sub 2}, resulted in similar total sugar yields about 40 g/100 g dry raw material. However, the fermentability of SO{sub 2}-impregnated material was better. This pretreatment resulted in the formation of inhibitors to the subsequent process steps, e.g. sugar and lignin degradation products. The glucose yield in the enzymatic hydrolysis stage was affected by various parameters such as enzyme loading, temperature, pH, residence time, substrate concentration, and agitation. To decrease the amount of fresh water used and thereby waste water produced, the sugar-rich prehydrolysate from the pretreatment step was included in the enzymatic hydrolysis of the solid fraction, resulting in a reduction in the cellulose conversion of up to 36%. Different prehydrolysate detoxification methods, such as treatment with Ca(OH){sub 2}, laccase, and fermentation using yeast, were investigated. The latter was shown to be very efficient. The amount of fresh water used can be further reduced by recycling various process streams. This was simulated experimentally in a bench-scale process. A reduction in fresh water demand of 50% was obtained without any further negative effects on either hydrolysis or fermentation.

  15. Hydrolysis of soy isoflavone conjugates using enzyme may underestimate isoflavone concentrations in tissue

    Hebron C. Chang; Myriam Laly; Melody Harrison; Thomas M. Badger


    Objective: To investigate the differences of using enzymatic hydrolysis and acid hydrolysis for identification and quantification of isoflavone aglycones from biomatrices. Methods: β-glucuronidase/sulfatase isolated from Helix pomatia for routine enzymatic hydrolysis or 6N HCl was used to release glucuronide and sulfate conjugates in the serum, urine and tissue samples. Profiles of soy isoflavones after enzymatic hydrolysis or acid hydrolysis in several tissues of rat fed with diets containing soy protein isolate were also compared using LC/MS and HPLC-ECD. Results: Acid hydrolysis released more aglycone than enzymatic digestion ( P <0.05) in liver tissue. The total genistein, daidzein and other metabolites were 20% to 60% lower in samples from enzymatic hydrolysis than in acid hydrolysis. Conclusion: These results indicated that unknown factors in tissues reduced the enzymatic hydrolytic efficiency for releasing isoflavone aglycones even in optimized condition. This would underestimate isoflavone tissue concentrations up to 60%.

  16. Modelling ethanol production from cellulose: separate hydrolysis and fermentation versus simultaneous saccharification and fermentation

    Drissen, R.E.T.; Maas, R.H.W.; Tramper, J.; Beeftink, H.H.


    In ethanol production from cellulose, enzymatic hydrolysis, and fermentative conversion may be performed sequentially (separate hydrolysis and fermentation, SHF) or in a single reaction vessel (simultaneous saccharification and fermentation, SSF). Opting for either is essentially a trade-off between

  17. Accelerated hydrolysis of substituted cellulose for potential biofuel production: kinetic study and modeling.

    Mu, Bingnan; Xu, Helan; Yang, Yiqi


    In this work, kinetics of substitution accelerated cellulose hydrolysis with multiple reaction stages was investigated to lay foundation for mechanism study and molecular design of substituting compounds. High-efficiency hydrolysis of cellulose is critical for cellulose-based bioethanol production. It is known that, substitution could substantially decrease activation energy and increase reaction rate of acidic hydrolysis of glycosidic bonds in cellulose. However, reaction kinetics and mechanism of the accelerated hydrolysis were not fully revealed. In this research, it was proved that substitution therefore accelerated hydrolysis only occurred in amorphous regions of cellulose fibers, and was a process with multiple reaction stages. With molar ratio of substitution less than 1%, the overall hydrolysis rate could be increased for around 10 times. We also quantified the relationship between the hydrolysis rate of individual reaction stage and its major influences, including molar ratio of substitution, activation energy of acidic hydrolysis, pH and temperature. PMID:26253917

  18. Study on Optimal Conditions of Alcalase Enzymatic Hydrolysis of Soybean Protein Isolate

    Yongsheng Ma; Xianhui Sun; Lintong Wang


    Soybean protein isolate was hydrolyzed to obtain soybean polypeptide solution using Alcalase as hydrolase. Degree of hydrolysis and the recovery rate of protein were used to characterize the soybean protein hydrolysis reaction result. Influence factors of soybean protein hydrolysis reaction including the substrate concentration, temperature, pH, enzyme concentration characterized by E/S (ratio of Enzyme and Substrate) and hydrolysis time were systematically studied with single factor and mult...

  19. Study on Hydrolysis Conditions of Flavourzyme in Soybean Polypeptide Alcalase Hydrolysate and Soybean Polypeptide Refining Process

    Yongsheng Ma; Lintong Wang; Xianhui Sun; Jianqiang Zhang; Junfeng Wang; Yue Li


    Soybean protein Alcalase hydrolysate was further hydrolyzed by adopting Flavourzyme as hydrolytic enzyme. The optimal hydrolysis conditions of Flavourzyme was that pH was 7.0 at temperature 50°C and E/S(ratio of enzyme and substrate) was 20LAPU/g. Bitterness value was reduced to 2 after Flavourzyme hydrolysis reaction in optimal hydrolysis conditions. The change of molecular weight distribution range from Alcalase hydrolysate to Flavourzyme hydrolysate was not obvious. DH (Degree of hydrolysi...

  20. Enzymatic hydrolysis of corn stalk in a hollow fiber ultrafiltration membrane reactor

    A hollow fiber ultrafiltration (UF) membrane reactor was set up to investigate the enzymatic hydrolysis of steam-exploded corn stalk. It was found that the hydrolysis rate, as well as the reducing sugar (RS) yield, could be markedly enhanced in the UF membrane reactor due to the continuous removal of inhibitory products. Compared with traditional batch hydrolysis, the hydrolysis rate and RS yield could increase 200% and 206%, respectively

  1. Influence of Incubation Conditions on Hydrolysis Efficiency and Iodine Enrichment in Baker’s Yeast

    Dolińska, Barbara; Zieliński, Michał; Dobrzański, Zbigniew; Chojnacka, Katarzyna; Opaliński, Sebastian; Ryszka, Florian


    The influence of incubation conditions, enzyme type, hydrolysis time, and potassium iodide concentration on hydrolysis and iodine enrichment were studied in supernatant and pellets of Saccharomyces cervisiae hydrolysates. The type of enzyme used and incubation time significantly influence hydrolysis efficiency and protein concentration in supernatant and pellet. The highest protein hydrolysis efficiency was obtained by 24-h incubation with papain. Significantly lower values were observed for ...

  2. Impact of the supramolecular structure of cellulose on the efficiency of enzymatic hydrolysis

    Peciulyte, Ausra; Karlström, Katarina; Larsson, Per Tomas; Olsson, Lisbeth


    Background The efficiency of enzymatic hydrolysis is reduced by the structural properties of cellulose. Although efforts have been made to explain the mechanism of enzymatic hydrolysis of cellulose by considering the interaction of cellulolytic enzymes with cellulose or the changes in the structure of cellulose during enzymatic hydrolysis, the process of cellulose hydrolysis is not yet fully understood. We have analysed the characteristics of the complex supramolecular structure of cellulose ...

  3. Comparison of Enzymatic Hydrolysis and Acid Hydrolysis of Sterol Glycosides from Foods Rich in Δ(7)-Sterols.

    Münger, Linda H; Jutzi, Sabrina; Lampi, Anna-Maija; Nyström, Laura


    In this study, we present the difference in sterol composition of extracted steryl glycosides (SG) hydrolyzed by either enzymatic or acid hydrolysis. SG were analyzed from foods belonging to the plant families Cucurbitaceae (melon and pumpkin seeds) and Amaranthaceae (amaranth and beetroot), both of which are dominated by Δ(7)-sterols. Released sterols were quantified by gas chromatography with a flame ionization detector (GC-FID) and identified using gas chromatography/mass spectrometry (GC-MS). All Δ(7)-sterols identified (Δ(7)-stigmastenyl, spinasteryl, Δ(7)-campesteryl, Δ(7)-avenasteryl, poriferasta-7,25-dienyl and poriferasta-7,22,25-trienyl glucoside) underwent isomerization under acidic conditions and high temperature. Sterols with an ethylidene or methylidene side chain were found to form multiple artifacts. The artifact sterols coeluted with residues of incompletely isomerized Δ(7)-sterols, or Δ(5)-sterols if present, and could be identified as Δ(8(14))-sterols on the basis of relative retention time, and their MS spectra as trimethylsilyl (TMS) and acetate derivatives. For instance, SG from melon were composed of 66% Δ(7)-stigmastenol when enzymatic hydrolysis was performed, whereas with acid hydrolysis only 8% of Δ(7)-stigmastenol was determined. The artifact of Δ(7)-stigmastenol coeluted with residual non-isomerized spinasterol, demonstrating the high risk of misinterpretation of compositional data obtained after acid hydrolysis. Therefore, the accurate composition of SG from foods containing sterols with a double bond at C-7 can only be obtained by enzymatic hydrolysis or by direct analysis of the intact SG. PMID:25757602


    Enzymatic hydrolysis of starch in corn is an important step that determines fermentation efficiency. Corn genetics, post harvest handling and process conditions are factors that affect starch hydrolysis. There is a lack of mathematical models for starch hydrolysis in the dry grind corn process tha...

  5. Analysis of Hydrolysis Reaction of N-Phosphorylphenylalanine by HPLC-ESI-MS/MS

    CAO Shu-Xia; ZHANG Jian-Chen; LIAO Xin-Cheng; ZHAO Yu-Fen


    @@ Hydrolysis procedure of N-diisopropyloxyphosphoryl phenylalanine (DIPP-Phe) has been studied by HPLCESI-MS. The hydrolysis products and intermediate were identified by HPLC-ESI-MS/MS. The results showed that (HO)(i-PrO)P(O)Phe was intermediate in the hydrolysis process.

  6. Switching Catalysis from Hydrolysis to Perhydrolysis in Pseudomonas fluorescens Esterase

    Yin, D.; Bernhardt, P; Morley, K; Jiang, Y; Cheeseman, J; Purpero, V; Schrag, J; Kazlauskas, R


    Many serine hydrolases catalyze perhydrolysis, the reversible formation of peracids from carboxylic acids and hydrogen peroxide. Recently, we showed that a single amino acid substitution in the alcohol binding pocket, L29P, in Pseudomonas fluorescens (SIK WI) aryl esterase (PFE) increased the specificity constant of PFE for peracetic acid formation >100-fold [Bernhardt et al. (2005) Angew. Chem., Int. Ed. 44, 2742]. In this paper, we extend this work to address the three following questions. First, what is the molecular basis of the increase in perhydrolysis activity? We previously proposed that the L29P substitution creates a hydrogen bond between the enzyme and hydrogen peroxide in the transition state. Here we report two X-ray structures of L29P PFE that support this proposal. Both structures show a main chain carbonyl oxygen closer to the active site serine as expected. One structure further shows acetate in the active site in an orientation consistent with reaction by an acyl-enzyme mechanism. We also detected an acyl-enzyme intermediate in the hydrolysis of {var_epsilon}-caprolactone by mass spectrometry. Second, can we further increase perhydrolysis activity? We discovered that the reverse reaction, hydrolysis of peracetic acid to acetic acid and hydrogen peroxide, occurs at nearly the diffusion limited rate. Since the reverse reaction cannot increase further, neither can the forward reaction. Consistent with this prediction, two variants with additional amino acid substitutions showed 2-fold higher k{sub cat}, but K{sub m} also increased so the specificity constant, k{sub cat}/K{sub m}, remained similar. Third, how does the L29P substitution change the esterase activity? Ester hydrolysis decreased for most esters (75-fold for ethyl acetate) but not for methyl esters. In contrast, L29P PFE catalyzed hydrolysis of {var_epsilon}-caprolactone five times more efficiently than wild-type PFE. Molecular modeling suggests that moving the carbonyl group closer to the

  7. The influence of solid/liquid separation techniques on the sugar yield in two-step dilute acid hydrolysis of softwood followed by enzymatic hydrolysis

    Galbe Mats; Monavari Sanam; Zacchi Guido


    Abstract Background Two-step dilute acid hydrolysis of softwood, either as a stand-alone process or as pretreatment before enzymatic hydrolysis, is considered to result in higher sugar yields than one-step acid hydrolysis. However, this requires removal of the liquid between the two steps. In an industrial process, filtration and washing of the material between the two steps is difficult, as it should be performed at high pressure to reduce energy demand. Moreover, the application of pressure...

  8. Conversion of rice straw to sugars by dilute-acid hydrolysis

    Hydrolysis of rice straw by dilute sulfuric acid at high temperature and pressure was investigated in one and two stages. The hydrolyses were carried out in a 10-l reactor, where the hydrolysis retention time (3-10 min), pressure (10-35 bar) and acid concentration (0-1%) were examined. Optimization of first stage hydrolysis is desirable to achieve the highest yield of the sugars from hemicellulose and also as a pretreatment for enzymatic hydrolysis. The results show the ability of first stage hydrolysis to depolymerize xylan to xylose with a maximum yield of 80.8% at hydrolysis pressure of 15 bar, 10 min retention time and 0.5% acid concentration. However, the yield of glucose from glucan was relatively low in first stage hydrolysis at a maximum of 25.8%. The solid residuals were subjected to further dilute-acid hydrolysis in this study. This second-stage hydrolysis without addition of the acid could not increase the yield of glucose from glucan beyond 26.6%. On the other hand, the best results of the hydrolysis were achieved, when 0.5% sulfuric acid was added prior to each stage in two-stage hydrolysis. The best results of the second stage of the hydrolysis were achieved at the hydrolysis pressure and the retention time of 30 bar and 3 min in the second stage hydrolysis, where a total of 78.9% of xylan and 46.6% of glucan were converted to xylose and glucose, respectively in the two stages. Formation of furfural and HMF were functions of the hydrolysis pressure, acid concentration, and retention time, whereas the concentration of acetic acid was almost constant at pressure of higher than 10 bar and a total retention time of 10 min

  9. Optimal extraction and hydrolysis of Chlorella pyrenoidosa proteins.

    Wang, Xiaoqin; Zhang, Xuewu


    In this study, for the first time, the applications of two new methods, ionic liquid and low-temperature high-pressure cell breakage methods, to the extraction of whole proteins in Chlorella pyrenoidosa cells were explored. Meanwhile, the comparison with three traditional methods was also made. The results indicated that the extraction rate for ionic liquid is only at moderate level, but the new low-temperature high-pressure cell breakage method can obviously increase the protein extraction rate up to 2- to 15-fold. Subsequently, the hydrolysis of the extracted proteins was conducted with three enzymes (papain, trypsin and alcalase). The data presented that the degree of hydrolysis for each enzyme under the optimal conditions is in the order of: alcalase (18.31%)>papain (14.33%)>trypsin (8.47%), demonstrating the potential of C. pyrenoidosa protein hydrolysates obtained here in nutritional supplement and medical foods. PMID:23117187

  10. Hydrolysis of Adiponitrile in Near-critical Water

    DUAN Pei-Gao; NIU Yan-Lei; WANG Yuan-Yuan; DAI Li-Yi


    Hydrolysis of adiponitrile (ADN) in near-critical water was successfully conducted in a batch reactor.Influences of m(AND)/m(water) ratio,temperature,time,m(AND)/m(additive) ratio,kind of additive and pressure on the yield of each product were investigated.Five compounds resulting from the hydrolysis of ADN,including 5-cyanovaleramide,adipamide,adipamic acid,adipic acid and trace of 5-cyanovaleric acid,were detected by high performance liquid chromatography.The results showed that change of ADN concentration and temperature had significant influences on the yields of adipamide,adipamic acid and adipic acid;time was the significant factor for the yield of 5-cyanovaleric acid;and the yield of 5-cyanovaleramide was more dependent on the ADN concentration.