Sample records for c-catalyzed sphingomyelin hydrolysis

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

    DEFF Research Database (Denmark)

    Zhang, Long; Liang, Shanshan; Hellgren, Lars


    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...... of sphingomyelin has been proven to be a feasible method to produce ceramide. In the membrane reactor constructed, the aqueous phase and the organic phase were separated by a membrane containing the immobilized enzyme, while the organic phasewas continuously circulated. Among the 10 selected membranes, the enzyme...... the low fixation level (9.4%). The optimal flow rate of the organic phase was 5 ml/min. High initial enzyme amount in the immobilization led to the decrease in the fixation level. Both the initial reaction rate and the specific activity of the enzyme increased with increasing enzyme loading, and slightly...

  2. Kinetic study of sphingomyelin hydrolysis for ceramide production

    DEFF Research Database (Denmark)

    Zhang, Long; Hellgren, Lars; Xu, Xuebing


    in 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...

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

    DEFF Research Database (Denmark)

    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...

  4. Sphingomyelinase D activity of brown recluse spider (Loxosceles reclusa) venom as studied by 31P-NMR: effects on the time-course of sphingomyelin hydrolysis. (United States)

    Merchant, M L; Hinton, J F; Geren, C R


    The time-course for the hydrolysis of the D linkage of chicken egg yolk sphingomyelin in a Triton X-100 mixed micelle and of lysophosphotidylcholine micelles, as catalyzed by brown recluse spider venom and brown recluse spider toxin, was followed by phosphorous-31 nuclear magnetic resonance spectroscopy. The overall rate of hydrolysis of sphingomyelin in mixed micelles was found to be an order of magnitude faster than the hydrolysis of lysophosphotidylcholine. Incorporation of lysophosphotidylcholine into mixed micelles with Triton X-100 inhibited the lipase activity of brown recluse spider venom and brown recluse spider venom toxin. The effects of increased rates of overall reaction were observed with increased temperature and also with decreased ionic strength. The presence of divalent calcium ions was found to be necessary for hydrolytic activity, but only in catalytic amounts (less than 1 mM).

  5. Enzymatic Production of Ceramide from Sphingomyelin

    DEFF Research Database (Denmark)

    Zhang, Long; Hellgren, Lars; Xu, Xuebing

    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...

  6. Inhibition of endothelial lipase activity by sphingomyelin in the lipoproteins. (United States)

    Yang, Peng; Belikova, Natalia A; Billheimer, Jeff; Rader, Daniel J; Hill, John S; Subbaiah, Papasani V


    Endothelial lipase (EL) is a major determinant of plasma HDL concentration, its activity being inversely proportional to HDL levels. Although it is known that it preferentially acts on HDL compared to LDL and VLDL, the basis for this specificity is not known. Here we tested the hypothesis that sphingomyelin, a major phospholipid in lipoproteins is a physiological inhibitor of EL, and that the preference of the enzyme for HDL may be due to low sphingomyelin/phosphatidylcholine (PtdCho) ratio in HDL, compared to other lipoproteins. Using recombinant human EL, we showed that sphingomyelin inhibits the hydrolysis of PtdCho in the liposomes in a concentration-dependent manner. While the enzyme showed lower hydrolysis of LDL PtdCho, compared to HDL PtdCho, this difference disappeared after the degradation of lipoprotein sphingomyelin by bacterial sphingomyelinase. Analysis of molecular species of PtdCho hydrolyzed by EL in the lipoproteins showed that the enzyme preferentially hydrolyzed PtdCho containing polyunsaturated fatty acids (PUFA) such as 22:6, 20:5, 20:4 at the sn-2 position, generating the corresponding PUFA-lyso PtdCho. This specificity for PUFA-PtdCho species was not observed after depletion of sphingomyelin by sphingomyelinase. These results show that sphingomyelin not only plays a role in regulating EL activity, but also influences its specificity towards PtdCho species.

  7. Enzymatic Modification of Sphingomyelin

    DEFF Research Database (Denmark)

    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-....... In present study, we have optimized the production of ceramide from sphingomyelin using Phospholipase C from Clostridium perfringens....

  8. Regulation of hepatic lipase activity by sphingomyelin in plasma lipoproteins. (United States)

    Yang, Peng; Subbaiah, Papasani V


    Hepatic lipase (HL) is an important enzyme in the clearance of triacylglycerol (TAG) from the circulation, and has been proposed to have pro-atherogenic as well as anti-atherogenic properties. It hydrolyzes both phospholipids and TAG of lipoproteins, and its activity is negatively correlated with HDL levels. Although it is known that HL acts preferentially on HDL lipids, the basis for this specificity is not known, since it does not require any specific apoprotein for activity. In this study, we tested the hypothesis that sphingomyelin (SM), whose concentration is much higher in VLDL and LDL compared to HDL, is an inhibitor of HL, and that this could explain the lipoprotein specificity of the enzyme. The results presented show that the depletion of SM from normal lipoproteins activated the HL roughly in proportion to their SM content. SM depletion stimulated the hydrolysis of both phosphatidylcholine (PC) and TAG, although the PC hydrolysis was stimulated more. In the native lipoproteins, HL showed specificity for PC species containing polyunsaturated fatty acids at sn-2 position, and produced more unsaturated lyso PC species. The enzyme also showed preferential hydrolysis of certain TAG species over others. SM depletion affected the specificity of the enzyme towards PC and TAG species modestly. These results show that SM is a physiological inhibitor of HL activity in lipoproteins and that the specificity of the enzyme towards HDL is at least partly due to its low SM content. Published by Elsevier B.V.

  9. Tales and mysteries of the enigmatic sphingomyelin synthase family

    NARCIS (Netherlands)

    Holthuis, J.C.M.|info:eu-repo/dai/nl/153674520; Luberto, C.


    In the last five years tremendous progress has been made toward the understanding of the mechanisms that govern sphingomyelin (SM) synthesis in animal cells. In line with the complexity of most biological processes, also in the case of SM biosynthesis, the more we learn the more enigmatic and finely

  10. Cardiovascular effects of sphingosine-1-phosphate and other sphingomyelin metabolites

    NARCIS (Netherlands)

    Alewijnse, Astrid E.; Peters, Stephan L. M.; Michel, Martin C.


    1 Upon various stimuli, cells metabolize sphingomyelin from the cellular plasma membrane to form sphingosylphosphorylcholine (SPC) or ceramide. The latter can be further metabolized to sphingosine and then sphingosine-1-phosphate (S1P). Apart from local formation, S1P and SPC are major constituents

  11. Subcellular targeting domains of sphingomyelin synthase 1 and 2. (United States)

    Yeang, Calvin; Ding, Tingbo; Chirico, William J; 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. 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. SMS1 is found exclusively in the Golgi at steady state, whereas SMS2 exists in the Golgi and plasma membrane. Conventional motifs responsible for protein targeting to the plasma membrane or Golgi are either not present in, or unique to, SMS1 and SMS2. In this study, we examined how SMS1 and SMS2 achieve their respective subcellular localization patterns. Brefeldin A treatment prevented SMS1 and SMS2 from exiting the ER, demonstrating that they transit through the classical secretory pathway. We created truncations and chimeras of SMS1 and SMS2 to define their targeting signals. We found that SMS1 contains a C-terminal Golgi targeting signal and that SMS2 contains a C-terminal plasma membrane targeting signal.

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

    Directory of Open Access Journals (Sweden)

    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.

  13. Sphingomyelin synthase overexpression increases cholesterol accumulation and decreases cholesterol secretion in liver cells

    Directory of Open Access Journals (Sweden)

    Li Yue


    Full Text Available Abstract Background Studies have shown that plasma high density lipoprotein cholesterol levels are negatively correlated with the development of atherosclerosis, whereas epidemiological studies have also shown that plasma sphingomyelin level is an independent risk factor for atherosclerosis. Methods To evaluate the relationship between cellular sphingomyelin level and cholesterol metabolism, we created two cell lines that overexpressed sphingomyelin synthase 1 or 2 (SMS1 or SMS2, using the Tet-off expression system. Results We found that SMS1 or SMS2 overexpression in Huh7 cells, a human hepatoma cell line, significantly increased the levels of intracellular sphingomyelin, cholesterol, and apolipoprotein A-I and decreased levels of apolipoprotein A-I and cholesterol in the cell culture medium, implying a defect in both processes. Conclusions Our findings indicate that the manipulation of sphingomyelin synthase activity could influence the metabolism of sphingomyelin, cholesterol and apolipoprotein A-I.

  14. Lovastatin reversed the enhanced sphingomyelin caused by 27-hydroxycholesterol in cultured vascular endothelial cells

    Directory of Open Access Journals (Sweden)

    Qi Zhou


    Full Text Available Statins have pleiotropic properties which are involved in inhibiting the thrombogenic response. In this study, the effects of lovastatin on two phospholipids, phosphatidylcholine and sphingomyelin, were studied in cultured endothelial cells in the presence of an oxysterol, 27-hydroxycholesterol. After the cells were cultured with 50 nM of lovastatin for 60 h, lovastatin was found to decrease the incorporation of [3H]choline into phosphatidylcholine and sphingomyelin, inhibited CTP: phosphocholine cytidylyltransferase (CT activity without altering the activity of sphingomyelin synthase and neutral sphingomyelinase. And lovastatin was not found to have a direct inhibitive effect on activity of CT. Exogenous mevalonic acid or cholesterol reversed the reduction of cholesterol concentration that was caused by lovastatin, but had no significant effect on the diminished [3H]sphingomyelin by lovastatin. The increase of [3H]sphingomyelin by 27-hydroxycholesterol was not detected in the presence of lovastatin. These findings suggest that (1 lovastatin can reduce sphingomyelin content by means of inhibiting phosphatidylcholine synthesis; and (2 The decrease in sphingomyelin is not related to the diminished cholesterol concentration or mevalonate-derived intermediates. This inhibitive effect of lovastatin on sphingomyelin may benefit cellular calcification caused by sphingomyelin.

  15. Comprehensive molecular motion capture for sphingomyelin by site-specific deuterium labeling. (United States)

    Matsumori, Nobuaki; Yasuda, Tomokazu; Okazaki, Hiroki; Suzuki, Takashi; Yamaguchi, Toshiyuki; Tsuchikawa, Hiroshi; Doi, Mototsugu; Oishi, Tohru; Murata, Michio


    Lipid rafts have attracted much attention because of their significant functional roles in membrane-associated processes. It is thought that sphingomyelin and cholesterol are essential for forming lipid rafts; however, their motion characteristics are not fully understood despite numerous studies. Here we show accurate local motions encompassing an entire sphingomyelin molecule, which were captured by measuring quadrupole splittings for 19 kinds of site-specifically deuterated sphingomyelins (that is, molecular motion capture of sphingomyelin). The quadrupole splitting profiles, which are distinct from those reported from perdeuterated sphingomyelins or simulation studies, reveal that cholesterol enhances the order in the middle parts of the alkyl chains more efficaciously than at the shallow positions. Comparison with dimyristoylphosphocholine bilayers suggests that cholesterol is deeper in sphingomyelin bilayers, which likely explains the so-called umbrella effect. The experiments also demonstrate that (i) the C2'-C3' bond predominantly takes the gauche conformation, (ii) the net ordering effect of cholesterol in sphingomyelin bilayers is not larger than that in phosphatidylcholine bilayers, (iii) cholesterol has no specific preference for the acyl or sphingosine chain, (iv) the acyl and sphingosine chains seem mismatched by about two methylene lengths, and (v) the motion of the upper regions of sphingomyelin chains is less temperature dependent than that of lower regions probably due to intermolecular hydrogen bond formation among SM molecules. These insights into the atomic-level dynamics of sphingomyelin provide critical clues to understanding the mechanism of raft formation.

  16. Consumption of olive oil has opposite effects on plasma total cholesterol and sphingomyelin concentrations in rats. (United States)

    Geelen, M J; Beynen, A C


    The hypothesis that olive-oil consumption alters plasma sphingomyelin concentrations and hepatic sphingomyelin metabolism was tested. Rats were fed on purified, high-cholesterol diets with either coconut fat or olive-oil (180 g/kg). In accordance with previous work, olive-oil v. coconut-fat consumption significantly elevated hepatic and total plasma cholesterol concentrations. During the course of the experiment, the concentration of plasma sphingomyelin rose in the coconut-fat group and remained constant in the olive-oil group. When compared with the coconut-fat-fed group, the plasma sphingomyelin levels were significantly lower in the olive-oil-fed group after 14 and 21 d of treatment. Dietary olive oil raised the amounts of cholesterol and sphingomyelin in the VLDL density region, and this change was associated with a reduction in the cholesterol and sphingomyelin contents of the LDL and HDL density ranges. Olive-oil consumption reduced the activity of serine palmitoyltransferase, while the activities of phosphatidylcholine:ceramide cholinephosphotransferase and phosphatidylethanolamine:ceramide ethanolaminephosphotransferase were left unchanged. Dietary olive oil also enhanced the activity of acidic sphingomyelinase, but not that of neutral sphingomyelinase. The present data indicate that dietary olive oil v. coconut fat has opposite effects on total plasma cholesterol and sphingomyelin concentrations. The lower plasma sphingomyelin levels observed in olive-oil-fed, as compared with coconut-fat-fed rats, may be explained by a simultaneous elevation and reduction in sphingomyelin catabolism and synthesis respectively, as based on the measured enzyme activities.

  17. Factors affecting longitudinal trajectories of plasma sphingomyelins: the Baltimore Longitudinal Study of Aging. (United States)

    Mielke, Michelle M; Bandaru, Veera Venkata Ratnam; Han, Dingfen; An, Yang; Resnick, Susan M; Ferrucci, Luigi; Haughey, Norman J


    Sphingomyelin metabolism has been linked to several diseases and to longevity. However, few epidemiological studies have quantified individual plasma sphingomyelin species (identified by acyl-chain length and saturation) or their relationship between demographic factors and disease processes. In this study, we determined plasma concentrations of distinct sphingomyelin species in 992 individuals, aged 55 and older, enrolled in the Baltimore Longitudinal Study of Aging. Participants were followed, with serial measures, up to 6 visits and 38 years (3972 total samples). Quantitative analyses were performed on a high-performance liquid chromatography-coupled electrospray ionization tandem mass spectrometer. Linear mixed models were used to assess variation in specific sphingomyelin species and associations with demographics, diseases, medications or lifestyle factors, and plasma cholesterol and triglyceride levels. We found that most sphingomyelin species increased with age. Women had higher plasma levels of all sphingomyelin species and showed steeper trajectories of age-related increases compared to men. African Americans also showed higher circulating sphingomyelin concentrations compared to Caucasians. Diabetes, smoking, and plasma triglycerides were associated with lower levels of many sphingomyelins and dihydrosphingomyelins. Notably, these associations showed specificity to sphingomyelin acyl-chain length and saturation. These results demonstrate that longitudinal changes in circulating sphingomyelin levels are influenced by age, sex, race, lifestyle factors, and diseases. It will be important to further establish the intra-individual age- and sex-specific changes in each sphingomyelin species in relation to disease onset and progression. © 2014 The Authors. Aging Cell published by the Anatomical Society and John Wiley & Sons Ltd.

  18. Miscibility Phase Diagrams of Giant Vesicles Containing Sphingomyelin (United States)

    Veatch, Sarah L.; Keller, Sarah L.


    Saturated sphingomyelin (SM) lipids are implicated in lipid rafts in cell plasma membranes. Here we use fluorescence microscopy to observe coexisting liquid domains in vesicles containing SM, an unsaturated phosphatidylcholine lipid (either DOPC or POPC), and cholesterol. We note similar phase behavior in a model membrane mixture without SM (DOPC/DPPC/Chol), but find no micron-scale liquid domains in membranes of POPC/PSM/Chol. We delineate the onset of solid phases below the miscibility transition temperature, and detail indirect evidence for a three-phase coexistence of one solid and two liquid phases.

  19. Local regulation of tooth mineralization by sphingomyelin phosphodiesterase 3. (United States)

    Khavandgar, Z; Alebrahim, S; Eimar, H; Tamimi, F; McKee, M D; Murshed, M


    Sphingomyelin phosphodiesterase 3 (Smpd3) encodes a membrane-bound enzyme that cleaves sphingomyelin to generate several bioactive metabolites. A recessive mutation called fragilitas ossium (fro) in the Smpd3 gene leads to impaired mineralization of bone and tooth extracellular matrix (ECM) in fro/fro mice. In teeth from fro/fro mice at various neonatal ages, radiography and light and electron microscopy showed delayed mantle dentin mineralization and a consequent delay in enamel formation as compared with that in control +/fro mice. These tooth abnormalities progressively improved with time. Immunohistochemistry showed expression of SMPD3 by dentin-forming odontoblasts. SMPD3 deficiency, however, did not affect the differentiation of these cells, as shown by osterix and dentin sialophosphoprotein expression. Using a transgenic mouse rescue model (fro/fro; Col1a1-Smpd3) in which Smpd3 expression is driven by a murine Col1a1 promoter fragment active in osteoblasts and odontoblasts, we demonstrate a complete correction of the tooth mineralization delays. In conclusion, analysis of these data demonstrates that Smpd3 expression in odontoblasts is required for tooth mineralization.

  20. Identification of mouse sphingomyelin synthase 1 as a suppressor of Bax-mediated cell death in yeast. (United States)

    Yang, Zhao; Khoury, Chamel; Jean-Baptiste, Gaël; Greenwood, Michael T


    We have identified mouse sphingomyelin synthase 1 as a novel suppressor of the growth inhibitory effect of heterologously expressed Bax. Yeast cells expressing sphingomyelin synthase 1 were also found to show an increased resistance to a variety of cytotoxic stimuli including hydrogen peroxide, osmotic stress and elevated temperature. Sphingomyelin synthase 1 functions by catalyzing the conversion of ceramide and phosphatidylcholine to sphingomyelin and diacylglycerol. Ceramide is an antiproliferative and proapoptotic sphingolipid whose level increases in response to a variety of stresses. Consistent with its biochemical function, yeast cells expressing sphingomyelin synthase 1 have an enhanced ability to grow in media containing the cell-permeable C2-ceramide analog as well as the ceramide precursor phytosphingosine. We also show that overexpression of AUR1, a potential yeast functional homolog of sphingomyelin synthase, also protects cells from osmotic stress. Taken together, these results suggest that sphingomyelin synthase 1 likely prevents cell death by counteracting stress-mediated accumulation of endogenous sphingolipids.

  1. Sphingomyelin exhibits greatly enhanced protection compared with egg yolk phosphatidylcholine against detergent bile salts

    NARCIS (Netherlands)

    Moschetta, A.; vanBerge-Henegouwen, G. P.; Portincasa, P.; Palasciano, G.; Groen, A. K.; van Erpecum, K. J.


    Inclusion of phosphatidylcholine within bile salt micelles protects against bile salt-induced cytotoxicity. In addition to phosphatidylcholine, bile may contain significant amounts of sphingomyelin, particularly under cholestatic conditions. We compared protective effects of egg yolk

  2. Acyl chain order and lateral domain formation in mixed phosphatidylcholine--sphingomyelin multilamellar and unilamellar vesicles. (United States)

    Lentz, B R; Hoechli, M; Barenholz, Y


    The phase behavior of mixtures of dimyristoylphosphatidylcholine (DMPC) with N-palmitoylsphingosinephosphorylcholine (C16SHP) has been investigated in both small unilamellar and large multilamellar vesicles. The steady-state fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH) has been used to detect temperature-induced structural changes in these membranes. In addition, electron microscopy has revealed vastly different fracture-face morphologies for large multilamellar vesicles "jet-frozen" from different temperatures. These data have been interpreted in terms of proposed phase diagrams for this lipid mixture. The shapes of the proposed phase diagrams have led us to conclude that phosphatidylcholine and sphingomyelin species of similar acyl chain length mix freely in both highly curved and uncurved bilayers, except at temperatures at which both lipids are in low-temperature, ordered phases. In addition, the similarity of these phase diagrams to phase diagrams for analogous mixtures of pure phosphatidylcholines suggested that sphingomyelin and phosphatidylcholine suggested that sphingomyelin and phosphatidylcholine species might substitute for each other in supporting the lamellar phase necessary for each other in supporting the lamellar phase necessary to cell membrane structure. Finally, the anisotropy of DPH fluorescence was found to be essentially invariant with sphingomyelin content at temperatures just above and below the solid--liquid phase separation in small unilamellar vesicles. This demonstrates that the sphingomyelin backbone, per se, does not order the membrane bilayer. These results are discussed in terms of the possible role of sphingomyelin in controlling acyl chain order within mammalian cell membranes.

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

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    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.

  4. The many faces (and phases) of ceramide and sphingomyelin II - binary mixtures. (United States)

    Fanani, María Laura; Maggio, Bruno


    A rather widespread idea on the functional importance of sphingolipids in cell membranes refers to the occurrence of ordered domains enriched in sphingomyelin and ceramide that are largely assumed to exist irrespective of the type of N-acyl chain in the sphingolipid. Ceramides and sphingomyelins are the simplest kind of two-chained sphingolipids and show a variety of species, depending on the fatty acyl chain length, hydroxylation, and unsaturation. Abundant evidences have shown that variations of the N-acyl chain length in ceramides and sphingomyelins markedly affect their phase state, interfacial elasticity, surface topography, electrostatics, and miscibility, and that even the usually conceived "condensed" sphingolipids and many of their mixtures may exhibit liquid-like expanded states. Their lateral miscibility properties are subtlety regulated by those chemical differences. Even between ceramides with different acyl chain length, their partial miscibility is responsible for a rich two-dimensional structural variety that impacts on the membrane properties at the mesoscale level. In this review, we will discuss the miscibility properties of ceramide, sphingomyelin, and glycosphingolipids that differ in their N-acyl or oligosaccharide chains. This work is a second part that accompanies a previous overview of the properties of membranes formed by pure ceramides or sphingomyelins, which is also included in this Special Issue.

  5. Purification, molecular cloning, and application of a novel sphingomyelin-binding protein (clamlysin) from the brackishwater clam, Corbicula japonica. (United States)

    Takara, Taketoshi; Nakagawa, Tetsuto; Isobe, Masami; Okino, Nozomu; Ichinose, Sachiyo; Omori, Akira; Ito, Makoto


    A novel sphingomyelin-binding protein (clamlysin) was purified from the foot muscle of a brackishwater clam, Corbicula japonica. The purified 24.8-kDa protein lysed sheep, horse and rabbit erythrocytes and the hemolytic activity was inhibited by sphingomyelin, but not other phospholipids or glycosphingolipids. The open reading frame of the clamlysin gene encoded a putative 26.9-kDa protein (clamlysin B) which showed high sequence similarity with the actinoporin family. A surface plasmon resonance assay confirmed that clamlysin B specifically bound to sphingomyelin. Furthermore, two cDNA variants of clamlysin, encoding putative 31.4 kDa (clamlysin A) and 11 kDa (clamlysin C) proteins, were isolated. Only the 31.4-kDa variant was found to exhibit sphingomyelin-binding activity. Clamlysin A and B, but not C, shared a sequence (domain II) conserved in all known sphingomyelin-binding proteins. Domain II fused with a glutathione S-transferase bound to sphingomyelin. Horse erythrocytes, mouse melanoma B16 and GM95 cells, and Chinese hamster ovary CHO-K1 cells, but not the same cells treated with bacterial sphingomyelinase, were immunostained with clamlysin B. These results indicate that clamlysin B binds to the sphingomyelin of living cells and thus would be useful as a molecular probe to detect sphingomyelin. Copyright © 2011 Elsevier B.V. All rights reserved.

  6. Lecithin/sphingomyelin ratio and lamellar body count for fetal lung maturity: a meta-analysis

    NARCIS (Netherlands)

    Besnard, Anouk E.; Wirjosoekarto, Soetinah A. M.; Broeze, Kimiko A.; Opmeer, Brent C.; Mol, Ben Willem J.


    To determine and compare the diagnostic accuracy of the lecithin/sphingomyelin (L/S) ratio and lamellar body count (LBC) in the prediction of neonatal respiratory distress syndrome (RDS). A systematic review was performed to identify studies comparing either the L/S ratio or the LBC with the

  7. The hydrolysis of polyimides (United States)

    Hoagland, P. D.; Fox, S. W.


    Thermal polymerization of aspartic acid produces a polysuccinimide (I), a chain of aspartoyl residues. An investigation was made of the alkaline hydrolysis of the imide rings of (I) which converts the polyimide to a polypeptide. The alkaline hydrolysis of polyimides can be expected to be kinetically complex due to increasing negative charge generated by carboxylate groups. For this reason, a diimide, phthaloyl-DL-aspartoyl-beta-alanine (IIA) was synthesized for a progressive study of the hydrolysis of polyimides. In addition, this diimide (IIA) can be related to thalidomide and might be expected to exhibit similar reactivity during hydrolysis of the phthalimide ring.

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

    DEFF Research Database (Denmark)

    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...... for crucial roles of alk-SMase in SM digestion and points to possible roles in regulating mucosal growth and alkaline phosphatase function....

  9. Sphingomyelin synthase 2 deficiency inhibits the induction of murine colitis-associated colon cancer. (United States)

    Ohnishi, Toshio; Hashizume, Chieko; Taniguchi, Makoto; Furumoto, Hidehiro; Han, Jia; Gao, Rongfen; Kinami, Shinichi; Kosaka, Takeo; Okazaki, Toshiro


    Sphingomyelin synthase 2 (SMS2) is the synthetic enzyme of sphingomyelin (SM), which regulates membrane fluidity and microdomain structure. SMS2 plays a role in LPS-induced lung injury and inflammation; however, its role in inflammation-mediated tumorigenesis is unclear. We investigated the effect of SMS2 deficiency on dextran sodium sulfate (DSS)-induced murine colitis and found inhibition of DSS-induced inflammation in SMS2-deficient (SMS2-/-) mice. DSS treatment induced a significant increase in ceramide levels, with a decrease of SM levels in SMS2-/- colon tissue, and demonstrated attenuation of the elevation of both inflammation-related gene expression and proinflammatory cytokines and chemokines, leukocyte infiltration, and MAPK and signal transducer and activator of transcription 3 activation. After undergoing transplantation of wild-type bone marrow, SMS2-/- mice also exhibited inhibition of DSS-induced inflammation in the colon, which suggested that SMS2 deficiency in bone marrow-derived immune cells was not involved in the inhibition of colitis. Finally, in an azoxymethane/DSS-induced cancer model, SMS2 deficiency significantly decreased tumor incidence in the colon. Our results demonstrate that SMS2 deficiency inhibits DSS-induced colitis and subsequent colitis-associated colon cancer via inhibition of colon epithelial cell-mediated inflammation; therefore, inhibition of SMS2 may be a potential therapeutic target for human colitis and colorectal cancer.-Ohnishi, T., Hashizume, C., Taniguchi, M., Furumoto, H., Han, J., Gao, R., Kinami, S., Kosaka, T., Okazaki, T. Sphingomyelin synthase 2 deficiency inhibits the induction of murine colitis-associated colon cancer. © FASEB.

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

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    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.

  11. Regulation of membrane KCNQ1/KCNE1 channel density by sphingomyelin synthase 1. (United States)

    Wu, Meikui; Takemoto, Makoto; Taniguchi, Makoto; Takumi, Toru; Okazaki, Toshiro; Song, Wen-Jie


    Sphingomyelin synthase (SMS) catalyzes the conversion of phosphatidylcholine and ceramide to sphingomyelin and diacylglycerol. We previously showed that SMS1 deficiency leads to a reduction in expression of the K(+) channel KCNQ1 in the inner ear (Lu MH, Takemoto M, Watanabe K, Luo H, Nishimura M, Yano M, Tomimoto H, Okazaki T, Oike Y, and Song WJ. J Physiol 590: 4029-4044, 2012), causing hearing loss. However, it remains unknown whether this change in expression is attributable to a cellular process or a systemic effect in the knockout animal. Here, we examined whether manipulation of SMS1 activity affects KCNQ1/KCNE1 currents in individual cells. To this end, we expressed the KCNQ1/KCNE1 channel in human embryonic kidney 293T cells and evaluated the effect of SMS1 manipulations on the channel using whole cell recording. Application of tricyclodecan-9-yl-xanthogenate, a nonspecific inhibitor of SMSs, significantly reduced current density and altered channel voltage dependence. Knockdown of SMS1 by a short hairpin RNA, however, reduced current density alone. Consistent with this, overexpression of SMS1 increased the current density without changing channel properties. Furthermore, application of protein kinase D inhibitors also suppressed current density without changing channel properties; this effect was nonadditive with that of SMS1 short hairpin RNA. These results suggest that SMS1 positively regulates KCNQ1/KCNE1 channel density in a protein kinase D-dependent manner. Copyright © 2016 the American Physiological Society.

  12. Role of sphingomyelin synthase in controlling the antimicrobial activity of neutrophils against Cryptococcus neoformans.

    Directory of Open Access Journals (Sweden)

    Asfia Qureshi


    Full Text Available The key host cellular pathway(s necessary to control the infection caused by inhalation of the environmental fungal pathogen Cryptococcus neoformans are still largely unknown. Here we have identified that the sphingolipid pathway in neutrophils is required for them to exert their killing activity on the fungus. In particular, using both pharmacological and genetic approaches, we show that inhibition of sphingomyelin synthase (SMS activity profoundly impairs the killing ability of neutrophils by preventing the extracellular release of an antifungal factor(s. We next found that inhibition of protein kinase D (PKD, which controls vesicular sorting and secretion and is regulated by diacylglycerol (DAG produced by SMS, totally blocks the extracellular killing activity of neutrophils against C. neoformans. The expression of SMS genes, SMS activity and the levels of the lipids regulated by SMS (namely sphingomyelin (SM and DAG are up-regulated during neutrophil differentiation. Finally, tissue imaging of lungs infected with C. neoformans using matrix-assisted laser desorption-ionization mass spectrometry (MALDI-MS, revealed that specific SM species are associated with neutrophil infiltration at the site of the infection. This study establishes a key role for SMS in the regulation of the killing activity of neutrophils against C. neoformans through a DAG-PKD dependent mechanism, and provides, for the first time, new insights into the protective role of host sphingolipids against a fungal infection.

  13. Effect of Alkali Metal Nitrates on the Ru/C-catalyzed Ring Hydrogenation of m-Xylylenediamine to 1,3-Cyclohexanebis(methylamine)

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Young Jin; Lee, Jae Hyeok; Kim, Hoon Sik [Kyung Hee Univ., Seoul (Korea, Republic of); Widyaya, Vania Tanda; Lee, Hyunjoo [Korea Institute of Science and Technology, Seoul (Korea, Republic of)


    Ru/C-catalyzed hydrogenation of m-xylylene diamine into 1,3-cyclohexanebis(methylamine) was greatly accelerated by the presence of LiNO{sub 3}, NaNO{sub 2}, or NaNO{sub 3}. It was found that the effect of the nitrate salt was significantly affected by the size of cation. The promoting effect of the nitrate salt increased with the decrease of the cation size: LiNO{sub 3} ∼ NaNO{sub 3} > KNO{sub 3} > CsNO{sub 3} >> [1-butyl-3-methylimidazolium]NO{sub 3}. XRD analysis of the recovered catalysts after the hydrogenation reactions showed that LiNO{sub 3} and NaNO{sub 3} were completely transformed into LiOH and NaOH, respectively, along with the evolution of NH{sub 3}, while KNO{sub 3} and CsNO{sub 3} remained unchanged.

  14. Mild and Robust Redox-Neutral Pd/C-Catalyzed Lignol β-O-4' Bond Cleavage Through a Low-Energy-Barrier Pathway. (United States)

    Galkin, Maxim V; Dahlstrand, Christian; Samec, Joseph S M


    A Pd/C catalyzed redox neutral C¢O bond cleavage of 2-aryloxy-1-arylethanols has been developed. The reactions are carried out at 80 °C, in air, using a green solvent system to yield the aryl ketones in near quantitative yields. Addition of catalytic amounts of a hydrogen source to the reaction mixture activates the catalyst to proceed through a low energy barrier pathway. Initial studies support a transfer hydrogenolysis reaction mechanism that proceeds through an initial dehydrogenation followed by an enol adsorption to Pd/C and a reductive C¢O bond cleavage. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. Enzymatic Hydrolysis of Lignocelluloses

    DEFF Research Database (Denmark)

    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...... source. By means of degenerate PCR, specific genes, homologous to the genes of previously classified glycoside hydrolases from CAZY database, are searched for in selected strains of Aspergillus sp., Trichoderma sp. and Penicillium sp. Both methods are anticipated to facilitate identification of target...

  16. Hydrolysis of biomass material (United States)

    Schmidt, Andrew J.; Orth, Rick J.; Franz, James A.; Alnajjar, Mikhail


    A method for selective hydrolysis of the hemicellulose component of a biomass material. The selective hydrolysis produces water-soluble small molecules, particularly monosaccharides. One embodiment includes solubilizing at least a portion of the hemicellulose and subsequently hydrolyzing the solubilized hemicellulose to produce at least one monosaccharide. A second embodiment includes solubilizing at least a portion of the hemicellulose and subsequently enzymatically hydrolyzing the solubilized hemicellulose to produce at least one monosaccharide. A third embodiment includes solubilizing at least a portion of the hemicellulose by heating the biomass material to greater than C. resulting in an aqueous portion that includes the solubilized hemicellulose and a water insoluble solids portion and subsequently separating the aqueous portion from the water insoluble solids portion. A fourth embodiment is a method for making a composition that includes cellulose, at least one protein and less than about 30 weight % hemicellulose, the method including solubilizing at least a portion of hemicellulose present in a biomass material that also includes cellulose and at least one protein and subsequently separating the solubilized hemicellulose from the cellulose and at least one protein.

  17. Catalytic carboxyester hydrolysis by diaminodiphenols

    Indian Academy of Sciences (India)


    Two diaminodiphenols, 1 and 2, have been examined as catalysts for the hydrolysis of 4- nitrophenyl acetate (NA) and 4-nitrophenylphosphate (NP) in aqueous-acetonitrile (25% acetonitrile v/v) media at 35ºC, I = 1·0 mol dm–3. The compound 1 enhances the hydrolysis rate of NA more than 105 times. Its catalytic efficiency ...

  18. Observing the mouse thyroid sphingomyelin under space conditions: a case study from the MDS mission in comparison with hypergravity conditions. (United States)

    Albi, E; Curcio, F; Spelat, R; Lazzarini, A; Lazzarini, R; Loreti, E; Ferri, I; Ambesi-Impiombato, F S


    This is a case report of apparent thyroid structural and functional alteration in a single mouse subjected to low Earth orbit spaceflight for 91 days. Histological examination of the thyroid gland revealed an increase in the average follicle size compared to that of three control animals and three animals exposed to hypergravity (2g) conditions. Immunoblotting analysis detected an increase in two thyroid gland enzymes, sphingomyelinase and sphingomyelin-synthase1. In addition, sphingomyelinase, an enzyme confined to the cell nucleus in the control animals, was found in the mouse exposed to hypogravity to be homogeneously distributed throughout the cell bodies. It represents the first animal observation of the influence of weightlessness on sphingomyelin metabolism.

  19. High-performance liquid chromatography of phosphatidylcholine and sphingomyelin with detection in the region of 200 nm. (United States)

    Jungalwala, F B; Evans, J E; McCluer, R H


    A sensitive method for the separation of phosphatidylcholine and sphingomyelin by high-performance liquid chromatographic analysis is described. The elution of the phospholipids from a microparticulate (10 mum) silica-gel chromatographic column was monitored with an ultraviolet spectromonitor at 203 nm. Acetonitrile/methanol/water (65:21:14, by vol.) was used as the solvent. It was shown by using synthetic phosphatidylcholines of knowm fatty acid composition and of varying degree of unsaturation that the absorption at 203 nm was primarily due to the isolated double bonds and the response measured varied with the degree of unsaturation. Approx. 1 nmol of phosphatidylcholine, containing at least one double bond per molecule, can be detected. The amounts of phosphatidylcholine and sphingomyelin could be determined by high-performance liquid chromatography and ultraviolet absorption if the apparent extinction coefficient of the material analyzed was established. Alternatively, peaks were collected and the phospholipids were determined by the analysis of phosphorus. The analysis of phosphatidylcholine and sphingomyelin present in the lipid extracts from animal tissues, blood and amniotic fluids were made without interference from other phospholipids or ultraviolet-absorbing material. The method described here is complementary to the high-performance liquid chromatographic method described previously for the analysis of ethanolamine-containing phosphoglycerides and serine-containing phosphoglycerides [Jungalwala, Turel, Evans and McCluer (1975) Biochem. J. 145, 517-526]. PMID:938476

  20. Sphingomyelin and phosphatidylcholine contrarily affect the induction of apoptosis in intestinal epithelial cells. (United States)

    Leucht, Katharina; Fischbeck, Anne; Caj, Michaela; Liebisch, Gerhard; Hartlieb, Eva; Benes, Petr; Fried, Michael; Humpf, Hans-Ulrich; Rogler, Gerhard; Hausmann, Martin


    The major alimentary sources for the plasma membrane lipid sphingomyelin (SM) are dairy products, eggs, and meat. We recently reported that the SM metabolite ceramide induces cathepsin D mediated apoptosis in murine intestinal epithelial cells (IECs) and increases inflammation in acute colitis. We investigated the impact of SM and phosphatidylcholine on apoptosis in human IECs and point out BH3-interacting death agonist (BID) as link between cathepsin D and apoptosis. HT-29 and isolated human IECs were stimulated with SM or phosphatidylcholine. SM treatment resulted in increased apoptosis. Phosphatidylcholine showed contrary effects. Western revealed higher amounts of cathepsin D and BID activation upon lipid stimulation. Western blotting revealed BID activation through SM in both an induced and a spontaneous mouse model of colitis. Dietary phospholipids may induce or abolish apoptosis in IECs and seem to play a role in the pathogenesis of inflammatory bowel diseases. This nutritional factor might be considered when evaluating the pathogenesis of inflammatory bowel diseases. Effects of SMase- and SM treatment on inflammation in dextran sulfate sodium induced animal models of colitis and in vitro experiments are discussed as controversial. Variable sources of SM, feeding techniques, and mouse strains might play a role. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Phospholipid lateral diffusion in phosphatidylcholine-sphingomyelin-cholesterol monolayers; effects of oxidatively truncated phosphatidylcholines. (United States)

    Parkkila, Petteri; Stefl, Martin; Olżyńska, Agnieszka; Hof, Martin; Kinnunen, Paavo K J


    Oxidative stress is involved in a number of pathological conditions and the generated oxidatively modified lipids influence membrane properties and functions, including lipid-protein interactions and cellular signaling. Brewster angle microscopy demonstrated oxidatively truncated phosphatidylcholines to promote phase separation in monolayers of 1-palmitoyl-2-oleoyl-sn-glycerol-3-phosphocholine (POPC), sphingomyelin (SM) and cholesterol (Chol). More specifically, 1-palmitoyl-2-azelaoyl-sn-glycero-3-phosphocholine (PazePC), was found to increase the miscibility transition pressure of the SM/Chol-phase. Lateral diffusion of lipids is influenced by a variety of membrane properties, thus making it a sensitive parameter to observe the coexistence of different lipid phases, for instance. The dependence on lipid lateral packing of the lateral diffusion of fluorophore-containing phospholipid analogs was investigated in Langmuir monolayers composed of POPC, SM, and Chol and additionally containing oxidatively truncated phosphatidylcholines, using fluorescence correlation spectroscopy (FCS). To our knowledge, these are the first FCS results on miscibility transition in ternary lipid monolayers, confirming previous results obtained using Brewster angle microscopy on such lipid monolayers. Wide-field fluorescence microscopy was additionally employed to verify the transition, i.e. the loss and reformation of SM/Chol domains. Copyright © 2014. Published by Elsevier B.V.

  2. Sustained activation of sphingomyelin synthase by 2-hydroxyoleic acid induces sphingolipidosis in tumor cells. (United States)

    Martin, Maria Laura; Liebisch, Gerhard; Lehneis, Stefan; Schmitz, Gerd; Alonso-Sande, María; Bestard-Escalas, Joan; Lopez, Daniel H; García-Verdugo, José Manuel; Soriano-Navarro, Mario; Busquets, Xavier; Escribá, Pablo V; Barceló-Coblijn, Gwendolyn


    The mechanism of action of 2-hydroxyoleic acid (2OHOA), a potent antitumor drug, involves the rapid and specific activation of sphingomyelin synthase (SMS), leading to a 4-fold increase in SM mass in tumor cells. In the present study, we investigated the source of the ceramides required to sustain this dramatic increase in SM. Through radioactive and fluorescent labeling, we demonstrated that sphingolipid metabolism was altered by a 24 h exposure to 2OHOA, and we observed a consistent increase in the number of lysosomes and the presence of unidentified storage materials in treated cells. Mass spectroscopy revealed that different sphingolipid classes accumulated in human glioma U118 cells after exposure to 2OHOA, demonstrating a specific effect on C16-, C20-, and C22-containing sphingolipids. Based on these findings, we propose that the demand for ceramides required to sustain the SMS activation (ca. 200-fold higher than the basal level) profoundly modifies both sphingolipid and phospholipid metabolism. As the treatment is prolonged, tumor cells fail to adequately metabolize sphingolipids, leading to a situation resembling sphingolipidosis, whereby cell viability is compromised.

  3. Sustained activation of sphingomyelin synthase by 2-hydroxyoleic acid induces sphingolipidosis in tumor cells1[S (United States)

    Martin, Maria Laura; Liebisch, Gerhard; Lehneis, Stefan; Schmitz, Gerd; Alonso-Sande, María; Bestard-Escalas, Joan; Lopez, Daniel H.; García-Verdugo, José Manuel; Soriano-Navarro, Mario; Busquets, Xavier; Escribá, Pablo V.; Barceló-Coblijn, Gwendolyn


    The mechanism of action of 2-hydroxyoleic acid (2OHOA), a potent antitumor drug, involves the rapid and specific activation of sphingomyelin synthase (SMS), leading to a 4-fold increase in SM mass in tumor cells. In the present study, we investigated the source of the ceramides required to sustain this dramatic increase in SM. Through radioactive and fluorescent labeling, we demonstrated that sphingolipid metabolism was altered by a 24 h exposure to 2OHOA, and we observed a consistent increase in the number of lysosomes and the presence of unidentified storage materials in treated cells. Mass spectroscopy revealed that different sphingolipid classes accumulated in human glioma U118 cells after exposure to 2OHOA, demonstrating a specific effect on C16-, C20-, and C22-containing sphingolipids. Based on these findings, we propose that the demand for ceramides required to sustain the SMS activation (ca. 200-fold higher than the basal level) profoundly modifies both sphingolipid and phospholipid metabolism. As the treatment is prolonged, tumor cells fail to adequately metabolize sphingolipids, leading to a situation resembling sphingolipidosis, whereby cell viability is compromised. PMID:23471028

  4. Sphingomyelin in High-Density Lipoproteins: Structural Role and Biological Function (United States)

    Martínez-Beamonte, Roberto; Lou-Bonafonte, Jose M.; Martínez-Gracia, María V.; Osada, Jesús


    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. PMID:23571495

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

    Directory of Open Access Journals (Sweden)

    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.

  6. A Lipid-Specific Toxin Reveals Heterogeneity of Sphingomyelin-Containing Membranes (United States)

    Ishitsuka, Reiko; Yamaji-Hasegawa, Akiko; Makino, Asami; Hirabayashi, Yoshio; Kobayashi, Toshihide


    Little is known about the heterogenous organization of lipids in biological membranes. Sphingomyelin (SM) is a major plasma membrane lipid that forms lipid domains together with cholesterol and glycolipids. Using SM-specific toxin, lysenin, we showed that in cultured epithelial cells the accessibility of the toxin to SM is different between apical and basolateral membranes. Apical membranes are highly enriched with glycolipids. The inhibitory role of glycolipids in the binding of lysenin to SM was confirmed by comparing the glycolipid-deficient mutant melanoma cell line with its parent cell. Model membrane experiments indicated that glycolipid altered the local density of SM so that the affinity of the lipid for lysenin was decreased. Our results indicate that lysenin recognizes the heterogenous organization of SM in biomembranes and that the organization of SM differs between different cell types and between different membrane domains within the same cell. Isothermal titration calorimetry suggests that lysenin binding to SM is presumably the result of a SM-lysenin complex formation of specific stoichiometry, thus supporting the idea of the existence of small condensed lipid complexes consisting of just a few lipid molecules in living cells. PMID:14695271

  7. Lecithin/sphingomyelin ratio and lamellar body count for fetal lung maturity: a meta-analysis. (United States)

    Besnard, Anouk E; Wirjosoekarto, Soetinah A M; Broeze, Kimiko A; Opmeer, Brent C; Mol, Ben Willem J


    To determine and compare the diagnostic accuracy of the lecithin/sphingomyelin (L/S) ratio and lamellar body count (LBC) in the prediction of neonatal respiratory distress syndrome (RDS). A systematic review was performed to identify studies comparing either the L/S ratio or the LBC with the occurrence of RDS published between January 1999 and February 2009. Two independent reviewers performed study selection and data extraction. For each study sensitivity and specificity were calculated. Summary receiver-operating characteristics (ROC) curves, assessing the diagnostic performance of both tests, were constructed. A subgroup analysis was performed to estimate the sensitivity and specificity of the various cut-off values. 13 studies were included. The ROC curves of the collected data illustrate that the LBC and L/S ratio perform equally well in the prediction of RDS. Comparison of the two summary ROC curves of each test indicates that the diagnostic performance of LBC might even have a slight advantage over L/S ratio. Due to the wide cut-off range it was not possible to define specific cut-off values with the best accuracy. We recommend replacing the L/S ratio as gold standard with the lamellar body count since the LBC is easy to perform, rapid, inexpensive, and available to all hospitals 24h per day. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

  8. Detailed Comparison of Deuterium Quadrupole Profiles between Sphingomyelin and Phosphatidylcholine Bilayers (United States)

    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

  9. Hydrolysis reactor for hydrogen production (United States)

    Davis, Thomas A.; Matthews, Michael A.


    In accordance with certain embodiments of the present disclosure, a method for hydrolysis of a chemical hydride is provided. The method includes adding a chemical hydride to a reaction chamber and exposing the chemical hydride in the reaction chamber to a temperature of at least about C. in the presence of water and in the absence of an acid or a heterogeneous catalyst, wherein the chemical hydride undergoes hydrolysis to form hydrogen gas and a byproduct material.

  10. Phase Behavior and Domain Size in Sphingomyelin-Containing Lipid Bilayers

    Energy Technology Data Exchange (ETDEWEB)

    Petruzielo, Robin S [Cornell University; Heberle, Frederick A [ORNL; Drazba, Paul [ORNL; Katsaras, John [ORNL; Feigenson, Gerald [Cornell University


    Membrane raft size measurements are crucial to understanding the stability and functionality of rafts in cells. The challenge of accurately measuring raft size is evidenced by the disparate reports of domain sizes, which range from nanometers to microns for the ternary model membrane system sphingomyelin (SM)/1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC)/cholesterol (Chol). Using F rster resonance energy transfer (FRET) and differential scanning calorimetry (DSC), we established phase diagrams for porcine brain SM (bSM)/dioleoyl-sn-glycero-3-phosphocholine (DOPC)/Chol and bSM/POPC/Chol at 15 and 25 C. By combining two techniqueswith different spatial sensitivities, namely FRET and small-angle neutron scattering (SANS),we have significantly narrowed the uncertainty in domain size estimates for bSM/POPC/Chol mixtures. Compositional trends in FRET data revealed coexisting domains at 15 and 25 C for bothmixtures, while SANS measurements detected no domain formation for bSM/POPC/Chol. Together these results indicate that liquid domains in bSM/POPC/Chol are between 2 and 7 nmin radius at 25 C: that is, domains must be on the order of the 2 6 nmF rster distance of the FRET probes, but smaller than the ~7 nm minimum cluster size detectable with SANS. However, for palmitoyl SM (PSM)/POPC/Chol at a similar composition, SANS detected coexisting liquid domains. This increase in domain size upon replacing the natural SMcomponent (which consists of amixture of chain lengths) with synthetic PSM, suggests a role for SM chain length in modulating raft size in vivo.

  11. Phase behavior and domain size in sphingomyelin-containing lipid bilayers. (United States)

    Petruzielo, Robin S; Heberle, Frederick A; Drazba, Paul; Katsaras, John; Feigenson, Gerald W


    Membrane raft size measurements are crucial to understanding the stability and functionality of rafts in cells. The challenge of accurately measuring raft size is evidenced by the disparate reports of domain sizes, which range from nanometers to microns for the ternary model membrane system sphingomyelin (SM)/1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC)/cholesterol (Chol). Using Förster resonance energy transfer (FRET) and differential scanning calorimetry (DSC), we established phase diagrams for porcine brain SM (bSM)/dioleoyl-sn-glycero-3-phosphocholine (DOPC)/Chol and bSM/POPC/Chol at 15 and 25°C. By combining two techniques with different spatial sensitivities, namely FRET and small-angle neutron scattering (SANS), we have significantly narrowed the uncertainty in domain size estimates for bSM/POPC/Chol mixtures. Compositional trends in FRET data revealed coexisting domains at 15 and 25°C for both mixtures, while SANS measurements detected no domain formation for bSM/POPC/Chol. Together these results indicate that liquid domains in bSM/POPC/Chol are between 2 and 7nm in radius at 25°C: that is, domains must be on the order of the 2-6nm Förster distance of the FRET probes, but smaller than the ~7nm minimum cluster size detectable with SANS. However, for palmitoyl SM (PSM)/POPC/Chol at a similar composition, SANS detected coexisting liquid domains. This increase in domain size upon replacing the natural SM component (which consists of a mixture of chain lengths) with synthetic PSM, suggests a role for SM chain length in modulating raft size in vivo. Copyright © 2012 Elsevier B.V. All rights reserved.

  12. A new cytolytic protein from the sea anemone Urticina crassicornis that binds to cholesterol- and sphingomyelin-rich membranes. (United States)

    Razpotnik, Andrej; Krizaj, Igor; Kem, William R; Macek, Peter; Turk, Tom


    A new pore-forming cytolytic protein was isolated from the Northern red sea anemone, Urticina crassicornis. Its biochemical properties were characterized and partial N-terminal amino acid sequence was determined. The cytolysin, named UcI, has a molecular mass of around 30kDa and lacks phospholipase A(2) activity. UcI lyses bovine erythrocytes at nanomolar concentrations. Hemolysis is a result of a colloid-osmotic shock caused by the opening of toxin-induced ionic pores and can be prevented by osmotic protectants of size >600Da. The functional radius of an average pore was estimated to be about 0.66nm. A more detailed study of the cytolytic activity of UcI was performed with lipid vesicles and monolayers. The toxin binds to monolayers and efficiently permeabilizes small lipid vesicles composed of sphingomyelin and cholesterol. However, the cytolytic activity is not prevented by preincubation with either pure cholesterol or sphingomyelin dispersions. We conclude that the presence of both sphingomyelin and cholesterol, key components of lipid rafts, greatly enhances toxin binding to membranes and probably facilitates pore formation. Alignment of the toxin partial amino acid sequence with sequences of cytolysins belonging to the actinoporin family reveals no sequence homology. We conclude that partial sequence of UcI resembles only the N-terminal part of UpI, a cytolytic protein isolated from a related sea anemone species, Urticina piscivora. The two proteins most probably belong to a separate family of sea anemone cytolysins that are worthy of further characterization.

  13. Effects of cations and cholesterol with sphingomyelin membranes investigated by high-resolution broadband sum frequency vibrational spectroscopy (United States)

    Zhang, Zhen; Feng, Rong-juan; Li, Yi-yi; Liu, Ming-hua; Guo, Yuan


    Sphingomyelin(SM) is specifically enriched in the plasma membrane of mammalian cells. Its molecular structure is compose by N-acyl-Derythro-sphingosylphosphorylcholine. The function of the SM related to membrane signaling and protein trafficking are relied on the interactions of the SM, cations, cholesterol and proteins. In this report, the interaction of three different nature SMs, cations and cholesterol at air/aqueous interfaces studied by high-resolution broadband sum frequency vibrational spectroscopy, respectively. Our results shed lights on understanding the relationship between SMs monolayer, cholesterol and Cations.

  14. Trivalent chromium induces autophagy by activating sphingomyelin phosphodiesterase 2 and increasing cellular ceramide levels in renal HK2 cells. (United States)

    Yang, Cheng-Lin; Chiou, Shiow-Her; Tai, Wei-Chun; Joseph, Nithila A; Chow, Kuan-Chih


    In this study, we examined the role of autophagy in the initiation of lipid increases in renal epithelial HK2 cells. We found that trivalent chromium [Cr(III)] induced autophagy by activating sphingomyelin phosphodiesterase 2 (SMPD2). SMPD2 increases levels of ceramide and other lipids. Confocal immunofluorescence microscopy showed that signals of ceramide overlapped with LC3, suggesting that ceramide might play an important role in the formation of autophagosome. In conclusion, our data indicate that Cr(III) induces autophagy via structural aberration of organelle membrane, in particular by the increase of lipid compositions in addition to autophagy-associated proteins. © 2017 Wiley Periodicals, Inc.

  15. Linchpin DNA-binding residues serve as go/no-go controls in the replication factor C-catalyzed clamp-loading mechanism. (United States)

    Liu, Juan; Zhou, Yayan; Hingorani, Manju M


    DNA polymerases depend on circular sliding clamps for processive replication. Clamps must be loaded onto primer-template DNA (ptDNA) by clamp loaders that open and close clamps around ptDNA in an ATP-fueled reaction. All clamp loaders share a core structure in which five subunits form a spiral chamber that binds the clamp at its base in a twisted open form and encloses ptDNA within, while binding and hydrolyzing ATP to topologically link the clamp and ptDNA. To understand how clamp loaders perform this complex task, here we focused on conserved arginines that might play a central coordinating role in the mechanism because they can alternately contact ptDNA or Walker B glutamate in the ATPase site and lie close to the clamp loader-clamp-binding interface. We mutated Arg-84, Arg-88, and Arg-101 in the ATPase-active B, C, and D subunits of Saccharomyces cerevisiae replication factor C (RFC) clamp loader, respectively, and assessed the impact on multiple transient events in the reaction: proliferating cell nuclear antigen (PCNA) clamp binding/opening/closure/release, ptDNA binding/release, and ATP hydrolysis/product release. The results show that these arginines relay critical information between the PCNA-binding, DNA-binding, and ATPase sites at all steps of the reaction, particularly at a checkpoint before RFC commits to ATP hydrolysis. Moreover, their actions are subunit-specific with RFC-C Arg-88 serving as an accelerator that enables rapid ATP hydrolysis upon contact with ptDNA and RFC-D Arg-101 serving as a brake that confers specificity for ptDNA as the correct substrate for loading PCNA. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  16. Cellulase hydrolysis of unsorted MSW

    DEFF Research Database (Denmark)

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


    of surfactant. Concurrently, the enzyme performance was analysed on pure cellulose in a solution of MSW wastewater. Results showed no effect of surfactant addition to the hydrolysis media as measured by viscosity and particle size distribution. MSW treatment wastewater was found to contain a high amount...

  17. The miscibility of milk sphingomyelin and cholesterol is affected by temperature and surface pressure in mixed Langmuir monolayers. (United States)

    Cheng, Ken; Ropers, Marie-Hélène; Lopez, Christelle


    The miscibility of milk sphingomyelin (milk-SM) and cholesterol was investigated in this study. The effect of different physical states of milk-SM on its interactions with cholesterol was determined by the recording of isotherms of compression of Langmuir films for temperatures above and below the gel to Lα phase transition of milk-SM (Tm∼34°C). For T=15°Cpressures of 10-15mN/m. For T=43°C>Tm, the milk-SM molecules were in a LE phase regardless of the surface pressure applied. A phase diagram pressure - milk-SM/cholesterol composition was established. This study demonstrated that both temperature and surface pressure affected the miscibility between the milk-SM and cholesterol. The strongest attractive forces (i.e. condensing effect) were identified for 30mol% cholesterol when the milk-SM was in the LE phase state. Copyright © 2016 Elsevier Ltd. All rights reserved.

  18. Enzymatic hydrolysis of plant extracts containing inulin

    Energy Technology Data Exchange (ETDEWEB)

    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).

  19. Kinetic modelling of enzymatic starch hydrolysis

    NARCIS (Netherlands)

    Bednarska, K.A.


    Kinetic modelling of enzymatic starch hydrolysis – a summary K.A. Bednarska The dissertation entitled ‘Kinetic modelling of enzymatic starch hydrolysis’ describes the enzymatic hydrolysis and kinetic modelling of liquefaction and saccharification of wheat starch.

  20. Enhancement of enzymatic adipyl-7-ADCA hydrolysis

    NARCIS (Netherlands)

    Schroën, C.G.P.H.; Kroon, P.J.; Vanderlaan, J.M.; Janssen, A.E.M.; Tramper, J.


    We studied enzymatic adipyl-7-ADCA hydrolysis as a new process for the production of 7-aminodeacetoxycephalosporanic acid (7-ADCA), one of the building blocks for cephalosporin antibiotics like cephalexin and cefadroxil. Adipyl-7-ADCA hydrolysis carried out with immobilised glutaryl acylase was

  1. Modeling the mechanisms of biological GTP hydrolysis

    DEFF Research Database (Denmark)

    Carvalho, Alexandra T.P.; Szeler, Klaudia; Vavitsas, Konstantinos


    in which GTP hydrolysis is activated and regulated is still a controversial topic and well-designed simulations can play an important role in resolving and rationalizing the experimental data. In this review, we discuss the contributions of computational biology to our understanding of GTP hydrolysis...

  2. Hydrolysis kinetics of dissolved polymer substrates

    NARCIS (Netherlands)

    Sanders, W.T.M.; Zeeman, G.; Lettinga, G.


    In this paper, the relation between the hydrolysis rate of dissolved polymer substrates and sludge concentration was investigated in two ways, viz. by laboratory experiments and by computer simulations. In the simulations, the hydrolysis of dissolved polymer components was regarded as a general

  3. Mass spectrometry images acylcarnitines, phosphatidylcholines, and sphingomyelin in MDA-MB-231 breast tumor models[S (United States)

    Chughtai, Kamila; Jiang, Lu; Greenwood, Tiffany R.; Glunde, Kristine; Heeren, Ron M. A.


    The lipid compositions of different breast tumor microenvironments are largely unknown due to limitations in lipid imaging techniques. Imaging lipid distributions would enhance our understanding of processes occurring inside growing tumors, such as cancer cell proliferation, invasion, and metastasis. Recent developments in MALDI mass spectrometry imaging (MSI) enable rapid and specific detection of lipids directly from thin tissue sections. In this study, we performed multimodal imaging of acylcarnitines, phosphatidylcholines (PC), a lysophosphatidylcholine (LPC), and a sphingomyelin (SM) from different microenvironments of breast tumor xenograft models, which carried tdTomato red fluorescent protein as a hypoxia-response element-driven reporter gene. The MSI molecular lipid images revealed spatially heterogeneous lipid distributions within tumor tissue. Four of the most-abundant lipid species, namely PC(16:0/16:0), PC(16:0/18:1), PC(18:1/18:1), and PC(18:0/18:1), were localized in viable tumor regions, whereas LPC(16:0/0:0) was detected in necrotic tumor regions. We identified a heterogeneous distribution of palmitoylcarnitine, stearoylcarnitine, PC(16:0/22:1), and SM(d18:1/16:0) sodium adduct, which colocalized primarily with hypoxic tumor regions. For the first time, we have applied a multimodal imaging approach that has combined optical imaging and MALDI-MSI with ion mobility separation to spatially localize and structurally identify acylcarnitines and a variety of lipid species present in breast tumor xenograft models. PMID:22930811

  4. Sphingomyelins and ceramides with VLCPUFAs are excluded from low-density raft-like domains in differentiating spermatogenic cells. (United States)

    Santiago Valtierra, Florencia X; Mateos, Melina V; Aveldaño, Marta I; Oresti, Gerardo M


    Rat spermatogenic cells contain sphingomyelins (SMs) and ceramides (Cers) with very long-chain PUFAs (VLCPUFAs) in nonhydroxylated (n-V) and 2-hydroxylated (h-V) forms. How these atypical species distribute among membrane fractions during differentiation was investigated here using a detergent-free procedure to isolate a small light raft-like low-density fraction and a large heavy fraction, mostly derived from the plasma membrane of spermatocytes, round spermatids, and late spermatids. The light fraction contained cholesterol, glycerophospholipids (GPLs), and SM with the same saturated fatty acids in all three stages. In the heavy fraction, as PUFA increased in the GPL and VLCPUFA in SM from spermatocytes to spermatids, the concentration of cholesterol was also augmented. The heavy fraction had mostly n-V SM in spermatocytes, but accumulated h-V SM and h-V Cer in spermatids. A fraction containing intracellular membranes had less SM and more Cer than the latter, but in both fractions SM and Cer species with h-V increased over species with n-V with differentiation. This accretion of h-V was consistent with the differentiation-dependent expression of fatty acid 2-hydroxylase (Fa2h), as it increased significantly from spermatocytes to spermatids. The non-raft region of the plasma membrane is thus the main target of the dynamic lipid synthesis and remodeling that is involved in germ cell differentiation. Copyright © 2017 by the American Society for Biochemistry and Molecular Biology, Inc.

  5. Orientation and Order of the Amide Group of Sphingomyelin in Bilayers Determined by Solid-State NMR (United States)

    Matsumori, Nobuaki; Yamaguchi, Toshiyuki; Maeta, Yoshiko; Murata, Michio


    Sphingomyelin (SM) and cholesterol (Chol) are considered essential for the formation of lipid rafts; however, the types of molecular interactions involved in this process, such as intermolecular hydrogen bonding, are not well understood. Since, unlike other phospholipids, SM is characterized by the presence of an amide group, it is essential to determine the orientation of the amide and its order in the lipid bilayers to understand the nature of the hydrogen bonds in lipid rafts. For this study, 1′-13C-2-15N-labeled and 2′-13C-2-15N-labeled SMs were prepared, and the rotational-axis direction and order parameters of the SM amide in bilayers were determined based on 13C and 15N chemical-shift anisotropies and intramolecular 13C-15N dipole coupling constants. Results revealed that the amide orientation was minimally affected by Chol, whereas the order was enhanced significantly in its presence. Thus, Chol likely promotes the formation of an intermolecular hydrogen-bond network involving the SM amide without significantly changing its orientation, providing a higher order to the SM amide. To our knowledge, this study offers new insight into the significance of the SM amide orientation with regard to molecular recognition in lipid rafts, and therefore provides a deeper understanding of the mechanism of their formation. PMID:26083921

  6. Hydrolysis of alkaline pretreated banana peel (United States)

    Fatmawati, A.; Gunawan, K. Y.; Hadiwijaya, F. A.


    Banana peel is one of food wastes that are rich in carbohydrate. This shows its potential as fermentation substrate including bio-ethanol. This paper presented banana peel alkaline pretreatment and enzymatic hydrolysis. The pretreatment was intended to prepare banana peel in order to increase hydrolysis performance. The alkaline pretreatment used 10, 20, and 30% w/v NaOH solution and was done at 60, 70 and 80°C for 1 hour. The hydrolysis reaction was conducted using two commercial cellulose enzymes. The reaction time was varied for 3, 5, and 7 days. The best condition for pretreatment process was one conducted using 30% NaOH solution and at 80°C. This condition resulted in cellulose content of 90.27% and acid insoluble lignin content of 2.88%. Seven-day hydrolysis time had exhibited the highest reducing sugar concentration, which was7.2869 g/L.

  7. Hemicellulose hydrolysis catalysed by solid acids

    NARCIS (Netherlands)

    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

  8. Process design and optimization of cellulose hydrolysis

    Energy Technology Data Exchange (ETDEWEB)

    Lindsey, R.R.; Wilke, C.R.


    The primary concern of this work is the economic optimization of a process for the hydrolysis of waste cellulosic material to fermentable sugars. Hydrolysis is performed enzymatically, utilizing the cellulase enzyme complex produced by Trichoderma viride. Using corn stover as a substrate, a system was designed to provide 14% hydrolyzate sugars (70% fermentable) at an estimated cost of 6.84 cents/pound of sugar, a 43% cost reduction over previous designs. Optimal residence time for hydrolysis was found to be 62 hours, resulting in a 34% conversion of raw material to sugars. Total fixed capital investment for the process is estimated to be $17.13 x 10/sup 6/. The kinetics of cellulose hydrolysis were modeled through the use of a modified Michaelis--Menten equation, making computer simulation of batch hydrolyses possible. Additional studies on the accessibility of cellulose were performed, and the feasibility of a counter-current processing scheme was investigated.

  9. Hydrolysis of isocyanic acid on SCR catalysts

    Energy Technology Data Exchange (ETDEWEB)

    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.

  10. Fusogenicity of Naja naja atra cardiotoxin-like basic protein on sphingomyelin vesicles containing oxidized phosphatidylcholine and cholesterol. (United States)

    Kao, Pei-Hsiu; Chen, Ying-Jung; Yang, Shin-Yi; Lin, Shinne-Ren; Hu, Wan-Ping; Chang, Long-Sen


    This study investigated the effect of oxidized phosphatidylcholine (oxPC) and cholesterol (Chol) on Naja naja atra cardiotoxin-like basic protein (CLBP)-induced fusion and leakage in sphingomyelin (SM) vesicles. Compared with those on PC/SM/Chol vesicles, CLBP showed a lower activity to induce membrane permeability but a higher fusogenicity on oxPC/SM/Chol vesicles. A reduction in inner-leaflet fusion elucidated that CLBP fusogenicity was not in parallel to its membrane-leakage activity on oxPC/SM/Chol vesicles. The lipid domain formed by Chol and SM supported CLBP fusogenicity on oxPC/SM/Chol vesicles, while oxPC altered the interacted mode of CLBP with oxPC/SM/Chol vesicles as evidenced by Fourier transform infrared spectra analyses and colorimetric phospholipid/polydiacetylene membrane assay. Although CLBP showed similar binding affinity with PC/SM/Chol and oxPC/SM/Chol vesicles, the binding capability of CLBP with PC/SM/Chol and oxPC/SM/Chol vesicles was affected differently by NaCl. This emphasized that CLBP adopted different membrane interaction modes upon binding with PC/SM/Chol and oxPC/SM/Chol vesicles. CLBP induced fusion in vesicles containing oxPC bearing the aldehyde group, and aldehyde scavenger methoxyamine abrogated the CLBP ability to induce oxPC/SM/Chol fusion. Taken together, our data indicate that Chol and oxPC bearing aldehyde group alter the CLBP membrane-binding mode, leading to fusogenicity promotion while reducing the membrane-damaging activity of CLBP.

  11. Sphingomyelin Liposomes Containing Soluble Leishmania major antigens Induced Strong Th2 Immune Response in BALB/c Mice

    Directory of Open Access Journals (Sweden)

    Omid Chavoshian


    Full Text Available   Objective(s: Soluble Leishmania antigens (SLA provide suitable protection against leishmaniasis in murine model when delivered by an appropriate delivery system. Liposomes have been shown to be suitable vaccine delivery systems against leishmaniasis, however, the phospholipase-A (PLA activity of SLA is a drawback to prepare a stable liposomal SLA. One strategy to overcome this problem might be using a lipid which is resistant to PLA activity of SLA such as sphingomyelin (SM. The aim of this study was to evaluate the effect of stable SM liposomes containing SLA on the immune response induced against leishmaniasis in BALB/c mice .   Materials and Methods: BALB/c mice were immunized subcutaneously, three times with 2-week intervals, with SLA, SM-liposome-SLA, empty liposome or buffer. As criteria for protection, footpads swelling at the site of challenge and foot parasite loads were assessed. The immune responses were also evaluated by determination of IgG subtypes and the level of IFN-γ and IL-4 in cultured splenocytes. Results: The group of mice receiving SM-liposome-SLA, showed a significant large footpad swelling, higher parasite burden in foot and higher IL-4 level compared to the group immunized with buffer. In terms of IgG and IgG isotypes, there was no significant difference between the mice receiving SM-liposome-SLA and the mice that received buffer. Moreover, the immune response induced by SM-liposome-SLA showed no significant difference compared with the one caused by SLA alone. Conclusion: It is concluded that SM-liposome-SLA is not an appropriate strategy to induce Th1 immune response and protect the mice against Leishmaniasis; however, SM-liposomes could be suitable vaccine delivery systems when a Th2 response is needed.

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

    Directory of Open Access Journals (Sweden)

    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.

  13. PC-PLC/sphingomyelin synthase activity plays a central role in the development of myogenic tone in murine resistance arteries. (United States)

    Mauban, Joseph R H; Zacharia, Joseph; Fairfax, Seth; Wier, Withrow Gil


    Myogenic tone is an intrinsic property of the vasculature that contributes to blood pressure control and tissue perfusion. Earlier investigations assigned a key role in myogenic tone to phospholipase C (PLC) and its products, inositol 1,4,5-trisphosphate (IP3) and diacylglycerol (DAG). Here, we used the PLC inhibitor, U-73122, and two other, specific inhibitors of PLC subtypes (PI-PLC and PC-PLC) to delineate the role of PLC in myogenic tone of pressurized murine mesenteric arteries. U-73122 inhibited depolarization-induced contractions (high external K(+) concentration), thus confirming reports of nonspecific actions of U-73122 and its limited utility for studies of myogenic tone. Edelfosine, a specific inhibitor of PI-PLC, did not affect depolarization-induced contractions but modulated myogenic tone. Because PI-PLC produces IP3, we investigated the effect of blocking IP3 receptor-mediated Ca(2+) release on myogenic tone. Incubation of arteries with xestospongin C did not affect tone, consistent with the virtual absence of Ca(2+) waves in arteries with myogenic tone. D-609, an inhibitor of PC-PLC and sphingomyelin synthase, strongly inhibited myogenic tone and had no effect on depolarization-induced contraction. D-609 appeared to act by lowering cytoplasmic Ca(2+) concentration to levels below those that activate contraction. Importantly, incubation of pressurized arteries with a membrane-permeable analog of DAG induced vasoconstriction. The results therefore mandate a reexamination of the signaling pathways activated by the Bayliss mechanism. Our results suggest that PI-PLC and IP3 are not required in maintaining myogenic tone, but DAG, produced by PC-PLC and/or SM synthase, is likely through multiple mechanisms to increase Ca(2+) entry and promote vasoconstriction. Copyright © 2015 the American Physiological Society.

  14. Hydrolytic gain during hydrolysis reactions : implications and correction procedures

    NARCIS (Netherlands)

    Marchal, L.M.; Tramper, J.


    Some of the structural parameters of starch (e.g. % beta- or gluco-hydrolysis) were influenced by the increase in mass during the hydrolysis reactions (hydrolytic gain). Procedures were derived to correct this apparent % of hydrolysis to actual % of hydrolysis. These analytically derived equations

  15. Kinetics of hydrolysis of meclofenoxate hydrochloride in human plasma. (United States)

    Yoshioka, S; Aso, Y; Uchiyama, M


    The kinetics of hydrolysis of meclofenoxate hydrochloride in human plasma have been compared with those of clofibrate. The hydrolysis rate in fractionated plasma was determined in the presence and absence of a plasma esterase inhibitor, tetraethyl pyrophosphate. The kinetic data indicated that clofibrate decomposed only by esterase-induced hydrolysis, which was inhibited by binding of clofibrate to plasma proteins. In contrast to clofibrate, meclofenoxate decomposed rapidly in human plasma via spontaneous hydrolysis as well as esterase-induced hydrolysis. The spontaneous hydrolysis appeared to be inhibited by some components present in the esterase fraction isolated from plasma, while no significant inhibition of the hydrolysis by protein binding was observed.

  16. Sterols Have Higher Affinity for Sphingomyelin than for Phosphatidylcholine Bilayers even at Equal Acyl-Chain Order (United States)

    Lönnfors, Max; Doux, Jacques P.F.; Killian, J. Antoinette; Nyholm, Thomas K.M.; Slotte, J. Peter


    The interaction between cholesterol and phospholipids in bilayer membranes is important for the formation and maintenance of membrane structure and function. However, cholesterol does not interact favorably with all types of phospholipids and, for example, prefers more ordered sphingomyelins (SMs) over phosphatidylcholines (PCs). The reason for this preference is not clear. Here we have studied whether acyl-chain order could be responsible for the preferred sterol interaction with SMs. Acyl-chain order was deduced from diphenylhexatriene anisotropy and from the deuterium order parameter obtained by 2H-NMR on bilayers made from either 14:0/14:0(d27)-PC, or 14:0(d27)-SM. Sterol/phospholipid interaction was determined from sterol bilayer partitioning. Cholestatrienol (CTL) was used as a fluorescence probe for cholesterol, because its relative membrane partitioning is similar to cholesterol. When CTL was allowed to reach equilibrium partitioning between cyclodextrins and unilamellar vesicles made from either 14:0/14:0-PC or 14:0-SM, the molar-fraction partitioning coefficient (Kx) was approximately twofold higher for SM bilayers than for PC bilayers. This was even the case when the temperature in the SM samples was raised to achieve equal acyl-chain order, as determined from 1,6-diphenyl-1,3,5-hexatriene (DPH) anisotropy and the deuterium order parameter. Although the Kx did increase with acyl-chain order, the higher Kx for SM bilayers was always evident. At equal acyl-chain order parameter (DPH anisotropy), the Kx was also higher for 14:0-SM bilayers than for bilayers made from either 14:0/15:0-PC or 15:0-/14:0-PC, suggesting that minor differences in chain length or molecular asymmetry are not responsible for the difference in Kx. We conclude that acyl-chain order affects the bilayer affinity of CTL (and thus cholesterol), but that it is not the cause for the preferred affinity of sterols for SMs over matched PCs. Instead, it is likely that the interfacial

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

    Directory of Open Access Journals (Sweden)

    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

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

    Directory of Open Access Journals (Sweden)

    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

  19. Hydrolysis and heat treatment of aluminum dust. (United States)

    López, F A; Peña, M C; López-Delgado, A


    Aluminum dust is a toxic and hazardous byproduct of Al remelting. The present research was performed to characterize and evaluate its behavior in water. The materials obtained by hydrolysis were also characterized, and the gases generated during the process were qualitatively analyzed. The effects of hydrolysis reaction time and temperature on the dust were also explored. The hydrolysis of Al dust is an exothermic reaction that gave rise to a solid composed of aluminum oxide, silicon oxide, and spinel (MgAl2O4). Most of the CH4, NH3, and SH2 gases generated were emitted immediately upon the start of the reaction, though their production continued for a long time. This slow reaction, which was moderately accelerated by temperature, led to the formation of a material less reactive than the untreated dust. On the other hand, heat treatment of the dust gave rise to an inert material composed of spinel, alumina, and magnesium and aluminum silicates.

  20. Additives enhancing enzymatic hydrolysis of lignocellulosic biomass. (United States)

    Rocha-Martín, Javier; Martinez-Bernal, Claudio; Pérez-Cobas, Yolanda; Reyes-Sosa, Francisco Manuel; García, Bruno Díez


    Linked to the development of cellulolytic enzyme cocktails from Myceliophthora thermophila, we studied the effect of different additives on the enzymatic hydrolysis yield. The hydrolysis of pretreated corn stover (PCS), sugar cane straw (PSCS) and microcrystalline cellulose (Avicel) was performed under industrial conditions using high solid loadings, limited mixing, and low enzyme dosages. The addition of polyethylene glycol (PEG4000) allowed to increase the glucose yields by 10%, 7.5%, and 32%, respectively in the three materials. PEG4000 did not have significant effect on the stability of the main individual enzymes but increased beta-glucosidase and endoglucanase activity by 20% and 60% respectively. Moreover, the presence of PEG4000 accelerated cellulase-catalyzed hydrolysis reducing up to 25% the liquefaction time. However, a preliminary economical assessment concludes that even with these improvements, a lower contribution of PEG4000 to the 2G bioethanol production costs would be needed to reach commercial feasibility. Copyright © 2017. Published by Elsevier Ltd.

  1. Enzymatic Modification of Sphingomyelin

    DEFF Research Database (Denmark)

    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...

  2. Enzymatic Modification of Sphingomyelin

    DEFF Research Database (Denmark)

    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...

  3. Enzymatic hydrolysis - present status and future developments

    Energy Technology Data Exchange (ETDEWEB)

    Linko, M.


    The environmental conditions for efficient cellulose and scylanate production with Trichoderma reesei have been studied. An interesting new aproach is cellulose production by Trichoderma reesei immobilized on K-carageenan. Hexoses and pentoses are produceed by the enzymatic hydrolysis of cellulose and hemicellulose and used for ethanol fermentation.


    Directory of Open Access Journals (Sweden)



    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.

  5. Starch facilitates enzymatic wheat gluten hydrolysis

    NARCIS (Netherlands)

    Hardt, N.A.; Boom, R.M.; Goot, van der A.J.


    Wheat gluten can be hydrolyzed by either using (vital) wheat gluten or directly from wheat flour. This study investigates the influence of the presence of starch, the main component of wheat, on enzymatic wheat gluten hydrolysis. Wheat gluten present in wheat flour (WFG) and vital wheat gluten (VWG)

  6. Microwave-assisted Weak Acid Hydrolysis of Proteins

    Directory of Open Access Journals (Sweden)

    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.


    Directory of Open Access Journals (Sweden)

    I. J. Baraldi

    Full Text Available Abstract Conventional production of instant coffee is based on solubilisation of polysaccharides present in roasted coffee. Higher process temperatures increase the solubilisation yield, but also lead to carbohydrate degradation and formation of undesirable volatile compounds. Enzymatic hydrolysis of roasted coffee is an alternative to minimize carbohydrate degradation. In this work, products obtained from thermal and enzymatic processes were compared in terms of carbohydrates and volatiles composition. Roasted coffee was extracted with water at 125 °C, and spent coffee was processed by thermal (180 °C or enzymatic hydrolysis. Enzymatic hydrolysis experiments were carried out at 50 °C using the commercial enzyme preparations Powercell (Prozyn, Galactomannanase (HBI-Enzymes, and Ultraflo XL (Novozymes. These formulations were previously selected from eleven different commercial enzyme preparations, and their main enzymatic activities included cellulase, galactomannanase, galactanase, and β-glucanase. Enzymatic hydrolysis yield was 18% (dry basis, similar to the extraction yield at 125 °C (20%, but lower than the thermal hydrolysis yield at 180 °C (28%. Instant coffee produced by enzymatic hydrolysis had a low content of undesirable volatile compounds and 21% (w/w of total carbohydrates. These results point to the enzymatic process as a feasible alternative for instant coffee production, with benefits including improved quality as well as reduced energy consumption.

  8. Short-time ultrasonication treatment in enzymatic hydrolysis of biomass (United States)

    Zengqian Shi; Zhiyong Cai; Siqun Wang; Qixin Zhong; Joseph J. Bozell


    To improve the conversion of enzymatic hydrolysis of biomass in an energy-efficient manner, two shorttime ultrasonication strategies were applied on six types of biomass with different structures and components. The strategies include pre-sonication before the hydrolysis and intermittent sonication during the ongoing hydrolysis. The microstructures of each type of...

  9. Cationic lipids (lipofectamine) and disturbance of cellular cholesterol and sphingomyelin distribution modulates gamma-secretase activity within amyloid precursor protein in vitro. (United States)

    Urmoneit, B; Turner, J; Dyrks, T


    To study beta-amyloid protein generation we expressed different amyloid precursor protein (APP) isoforms in the human neuroblastoma cell line SY5Y (for details see (1)). Treatment with lipofectamine, an cationic lipid for eucaryotic cell transfection, inhibits gamma-secretase activity and stimulates the physiological APP cleavage by alpha-secretase activity. Beside the MDL inhibitor (2), this is the second agent that shows modulation of gamma-secretase activity in vitro. Further, we show that disturbance of cellular cholesterol and sphingomyelin distribution in transfected SY5Y cells results in an overproduction of beta-amyloid protein. This provides experimental evidence that membrane instability influenced the proteolytic activity of gamma-secretase within the APP molecule.

  10. Electrospray ionization mass spectrometry for the hydrolysis complexes of cisplatin : Implications for the hydrolysis process of platinum complexes

    NARCIS (Netherlands)

    Xie, Feifan; Colin, Pieter; Van Bocxlaer, Jan

    Non-enzyme-dependent hydrolysis of the drug cisplatin is important for its mode of action and toxicity. However, up until today, the hydrolysis process of cisplatin is still not completely understood. In the present study, the hydrolysis of cisplatin in an aqueous solution was systematically

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

    NARCIS (Netherlands)

    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

  12. Enzymatic hydrolysis of corn bran arabinoxylan

    DEFF Research Database (Denmark)

    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...


    Directory of Open Access Journals (Sweden)



    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.


    Directory of Open Access Journals (Sweden)



    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

  15. Effects of enzymatic hydrolysis on lentil allergenicity. (United States)

    Cabanillas, Beatriz; Pedrosa, Mercedes M; Rodríguez, Julia; González, Angela; Muzquiz, Mercedes; Cuadrado, Carmen; Crespo, Jesús F; Burbano, Carmen


    Enzymatic hydrolysis and further processing are commonly used to produce hypoallergenic dietary products derived from different protein sources, such as cow's milk. Lentils and chickpeas seem to be an important cause of IgE-mediated hypersensitivity in the Mediterranean area and India. Some studies have investigated the effects of enzymatic treatments on the in vitro immunological reactivity of members of the Leguminosae family, such as soybean, chickpea, lentil, and lupine. Nevertheless, there are only a few studies carried out to evaluate the effect on IgE reactivity of these food-hydrolysis products with sera from patients with well-documented allergy to these foods. In this study, lentil protein extract was hydrolyzed by sequential action of an endoprotease (Alcalase) and an exoprotease (Flavourzyme). Immunoreactivity to raw and hydrolyzed lentil extract was evaluated by means of IgE immunoblotting and ELISA using sera from five patients with clinical allergy to lentil. The results indicated that sequential hydrolysis of lentil results in an important proteolytic destruction of IgE-binding epitopes shown by in vitro experiments. However, some allergenic proteins were still detected by sera from four out of five patients in the last step of sequential hydrolyzation.

  16. Palm date fibers: analysis and enzymatic hydrolysis. (United States)

    Shafiei, Marzieh; Karimi, Keikhosro; Taherzadeh, Mohammad J


    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.

  17. Palm Date Fibers: Analysis and Enzymatic Hydrolysis

    Directory of Open Access Journals (Sweden)

    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.

  18. Water Availability as a Measure of Cellulose Hydrolysis Efficiency

    DEFF Research Database (Denmark)

    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...... availability is vital for efficient hydrolysis, especially at high dry matter content where water availability is low. At high dry matter content, cellulase activity changes water interactions with biomass, affecting the water mobility. While swelling and fiber loosening also take place during hydrolysis...... 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...

  19. Degradation of Opioids and Opiates During Acid Hydrolysis Leads to Reduced Recovery Compared to Enzymatic Hydrolysis. (United States)

    Sitasuwan, Pongkwan; Melendez, Cathleen; Marinova, Margarita; Mastrianni, Kaylee R; Darragh, Alicia; Ryan, Emily; Lee, L Andrew


    Drug monitoring laboratories utilize a hydrolysis process to liberate the opiates from their glucuronide conjugates to facilitate their detection by tandem mass spectrometry (MS). Both acid and enzyme hydrolysis have been reported as viable methods, with the former as a more effective process for recovering codeine-6-glucuronide and morphine-6-glucuronide. Here, we report concerns with acid-catalyzed hydrolysis of opioids, including a significant loss of analytes and conversions of oxycodone to oxymorphone, hydrocodone to hydromorphone and codeine to morphine. The acid-catalyzed reaction was monitored in neat water and patient urine samples by liquid chromatography-time-of-flight and tandem MS. These side reactions with acid hydrolysis may limit accurate quantitation due to loss of analytes, possibly lead to false positives, and poorly correlate with pharmacogenetic profiles, as cytochrome P450 enzyme (CYP2D6) is often involved with oxycodone to oxymorphone, hydrocodone to hydromorphone and codeine to morphine conversions. Enzymatic hydrolysis process using the purified, genetically engineered β-glucuronidase (IMCSzyme(®)) addresses many of these concerns and demonstrates accurate quantitation and high recoveries for oxycodone, hydrocodone, oxymorphone and hydromorphone. © The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email:

  20. Organosolv pretreatment for enzymatic hydrolysis of poplars: I. enzyme hydrolysis of cellulosic residues

    Energy Technology Data Exchange (ETDEWEB)

    Chum, H.L.; Johnson, D.K.; Black, S.; Baker, J.; Grohmann, K.; Sarkanen, K.V.; Wallace, K.; Schroeder, H.A.


    Aspen (Populus tremuloides) and black cottonwood (Populus trichocarpa) organosolv pulps produced in a wide range of solvent composition (between 30 and 70% by volume of methanol) and catalysts (H/sub 2/SO/sub 4/ and H/sub 3/PO/sub 4/) such that the cooking liquor pH less than or equal to 3 are easily digested by enzymes. The total yields of hydrolysis residues (pulps) are in the 40-60% range; the acid-catalyzed delignification followed by enzyme hydrolysis can generate 70-88% of the original six-carbon sugars contained in the wood. Glucomannan and arabinogalactan are dissolved in to the pulping liquor in the pH range of 2-4.5. Lower pH (less than or equal to 3) leads to additional solubilization of six-carbon sugars. These sugars may be fermented directly. From the insoluble hydrolysis residues, 36-41% conversions of wood into fermentable sugars were obtained after enzyme hydrolysis; the starting feedstocks contain 50.8 and 46.6% hexosans, respectively, for aspen and black cottonwood. The kinetics of enzymatic hydrolysis of cellulose can be formally treated as two simultaneous pseudo-first-order reactions in which fast and slow hydrolysis of cellulose occur. Correlations between the glucan digestibility and the effect of the pretreatment have been made. The higher residual xylan content reduces the amount of the rapidly hydrolyzable glucan fraction and lowers the glucan digestibility. The proposed simple kinetic treatment is very helpful in assessing the effect of the pretreatment on pulp enzyme hydrolyzability.

  1. Synergy between cellulases and pectinases in the hydrolysis of hemp. (United States)

    Zhang, Junhua; Pakarinen, Annukka; Viikari, Liisa


    The impact of pectinases in the hydrolysis of fresh, steam-exploded and ensiled hemp was investigated and the synergy between cellulases, pectinases and xylanase in the hydrolysis was evaluated. About half; 59.3% and 46.1% of pectin in the steam-exploded and ensiled hemp, respectively, could be removed by a low dosage of pectinases used. Pectinases were more efficient than xylanase in the hydrolysis of fresh and ensiled hemp whereas xylanase showed higher hydrolytic efficiency than the pectinase preparation used in the hydrolysis of steam-exploded hemp. Clear synergistic action between cellulases and xylanase could be observed in the hydrolysis of steam-exploded hemp. Supplementation of pectinase resulted in clear synergism with cellulases in the hydrolysis of all hemp substrates. Highest hydrolysis yield of steam-exploded hemp was obtained in the hydrolysis with cellulases and xylanase. In the hydrolysis of ensiled hemp, the synergistic action between cellulases and pectinases was more obvious for efficient hydrolysis. Copyright © 2012 Elsevier Ltd. All rights reserved.

  2. Pretreatment and enzymatic hydrolysis of lignocellulosic biomass (United States)

    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

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

    DEFF Research Database (Denmark)

    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 dem......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...

  4. The Hydrolysis of Carbonyl Sulfide at Low Temperature: A Review (United States)

    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

  5. Enzymatic hydrolysis of lactose of whey permeate

    Directory of Open Access Journals (Sweden)

    Karina Nascimento de Almeida


    Full Text Available The whey permeate is the residual of the concentration process of the whey proteins by ultrafiltration method. It contains important nutrients such as lactose, minerals and some proteins and lipids. It is without an ending industrial waste that causes serious damage to the environment. For its full use the lactose must be hydrolyzed to enable its consumption by intolerant people. The enzymatic hydrolysis by lactase (β-galactosidase of Kluyveromyces lactis yeast is a safe method that does not compromise the integrity of other nutrients, enabling further use of the permeate as a raw material. This study aimed to perform tests of enzymatic hydrolysis of lactose in whey permeate formulations in a concentration of 0.2%, 0.7% and 1% at 30, 60 and 90 minutes with pH 6.3 medium and 37 °C. The reactions were monitored by high performance liquid chromatography which showed that the enzyme concentration of 0.7% at time 30 minutes formulations became safe for consumption by lactose intolerant people, according to minimum levels established by law.

  6. Design and test of urea hydrolysis reactor for vehicle

    Directory of Open Access Journals (Sweden)

    Qi Zhanfeng


    Full Text Available Ammonia production technology of urea-SCR system for vehicle is mainly used in pyrolysis. The reaction is complex, and there are some side effects. So a kind of urea hydrolysis device for vehicle is designed. Based on the in-depth analysis of ammonia production technology’s mechanism of urea hydrolysis for vehicle, the modified extended UNIQUAC equation and PHS equation of state were used to solve the thermodynamic model, and the experimental verification was carried out on the JX493ZLQ3 diesel engine. The results show that the design of urea hydrolysis reactor is in agreement with the experimental results. In the environment of urea hydrolysis, the higher the hydrolysis temperature, the faster the urea hydrolysis and the faster the production of ammonia. Under the same conditions, the hydrolysis rate of urea was the same, the higher the initial quality, the more ammonia produced. The method is suitable for the ammonia production technology of urea hydrolysis for vehicle and can be used for the formation of urea hydrolysis SCR system.

  7. Effect of gelatinization and hydrolysis conditions on the selectivity of starch hydrolysis with alpha-amylase from Bacillus licheniformis. (United States)

    Baks, Tim; Bruins, Marieke E; Matser, Ariette M; Janssen, Anja E M; Boom, Remko 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 alpha-amylase from Bacillus licheniformis were compared. Suspensions of native starch or starch gelatinized at different conditions either with or without enzyme were hydrolyzed. During hydrolysis, the oligosaccharide concentration, the dextrose equivalent, and the enzyme activity were determined. We found that the hydrolyzate composition was affected by the type of starch pretreatment and the enzyme addition point but that it was just minimally affected by the pressure applied during hydrolysis, as long as gelatinization was complete. The differences between hydrolysis of thermally gelatinized, high-pressure gelatinized, and native starch were explained by considering the granule structure and the specific surface area of the granules. These results show that the hydrolyzate composition can be influenced by choosing different process sequences and conditions.

  8. [Urinary oestriol excretion and the lecithin/sphingomyelin ratio in amniotic fluid in Bogota (2600 m) I. Normal pregnancy (author's transl)]. (United States)

    Sobrevilla, L A; Cristina Peña, M; Jaramillo, R


    We have studied 22 pregnancies in order to establish normal values for the urinary oestriol excretion in Bogotá, a city 2600 metres above sea level. The study subjects were normal pregnant women attending the prenatal clinic of the Hospital San José de Bogotá, and belong to a racially mixed community of medium to low socio-economic level. In the study, new born weight was found low (mean +/- SEM 2.97 +/-0.06 kg) while placental weight was high (0.6 +/- 0.02 kg) with a high placenta/newborn ratio. Maternal hemoglobin was elevated (12.8 +/- 0.2g/100 ml) reflecting the effect of altitude. In 66 determinations, oestriol excretion was more than 4mg/24 hours from week 31 to 36 and of more than 5 mg/24 hours from week 35 to 40. The decreased excretion of oestriol most likely reflects impaired intrauterine fetal growth, and is probably related to nutrional, racial and socio-economic factors as well as to the altitude. In five normal term pregnancies studied, the lecithin/sphingomyelin ratio was of 2 or more and amniotic fluid creatinine was also elevated, indicating maturity of the pulmonary and renal enzyme systems of the fetus.

  9. Fermentable sugars by chemical hydrolysis of biomass (United States)

    Binder, Joseph B.; Raines, Ronald T.


    Abundant plant biomass has the potential to become a sustainable source of fuels and chemicals. Realizing this potential requires the economical conversion of recalcitrant lignocellulose into useful intermediates, such as sugars. We report a high-yielding chemical process for the hydrolysis of biomass into monosaccharides. Adding water gradually to a chloride ionic liquid-containing catalytic acid leads to a nearly 90% yield of glucose from cellulose and 70–80% yield of sugars from untreated corn stover. Ion-exclusion chromatography allows recovery of the ionic liquid and delivers sugar feedstocks that support the vigorous growth of ethanologenic microbes. This simple chemical process, which requires neither an edible plant nor a cellulase, could enable crude biomass to be the sole source of carbon for a scalable biorefinery. PMID:20194793

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

    DEFF Research Database (Denmark)

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


    and 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...... 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 tests...

  11. Kinetics of ptaquiloside hydrolysis in aqueous solution

    DEFF Research Database (Denmark)

    Ayala-Luis, Karina B.; Bildsøe Hansen, Pernille; Rasmussen, Lars Holm


    of the toxin, a full understanding of the PTA degradation in aqueous environments is important. The kinetics of PTA hydrolysis was examined at 22C in aqueous buffered solutions (pH 2.88–8.93). The reaction was found to follow first-order kinetics with respect to PTA at all pH and temperature conditions. At p.......46 is strongly dependent on temperature, with an activation energy of 74.4 (2.6) kJ mol1. Stoichiometric calculations, reaction kinetics, and ultraviolet–visible spectrophotometry strongly indicates the formation of an intermediary compound at pH 5.07 and 6.07 via a mechanism comprising two first...

  12. The kinetics of hydrolysis of acetylsalicylic acid (Aspirin) in different ...

    African Journals Online (AJOL)

    The kinetics of hydrolysis of Acetylsalicylic acid (Aspirin) to salicylic acid was followed by the direct spectrophotometric measurement of the amount of salicylic acid produced with time. Salicylic acid was complexed with ferric ion giving a characteristic purple colour (λlm 523nm). The kinetics of hydrolysis was found to follow ...

  13. Factors Impeding Enzymatic Wheat Gluten Hydrolysis at High Solid Concentrations

    NARCIS (Netherlands)

    Hardt, N.A.; Janssen, A.E.M.; Boom, R.M.; Goot, van der A.J.


    Enzymatic wheat gluten hydrolysis at high solid concentrations is advantageous from an environmental and economic point of view. However, increased wheat gluten concentrations result in a concentration effect with a decreased hydrolysis rate at constant enzyme-to-substrate ratios and a decreased

  14. A comparison of acidic and enzymatic hydrolysis of rutin | Wang ...

    African Journals Online (AJOL)

    Rutin and its hydrolysis products (isoquercitrin and quercetin) are widely used as important materials in food and pharmaceutical industry. In this study, the effects of various acids and enzymes as catalysts on the hydrolysis reaction of rutin were studied. In comparison with acidic and enzymatic catalysis of rutin, the research ...

  15. Production of value added materials by subcritical water hydrolysis ...

    African Journals Online (AJOL)

    The aim of this study was the determination of the best experimental conditions for the production of useful materials such as amino acids by subcritical water hydrolysis from supercritical carbon dioxide extracted krill residues and to compare the results with raw krill. Subcritical water hydrolysis efficiency from raw and ...

  16. Photodegradation and Hydrolysis of Chlorpyrifos in Aqueous Systems

    African Journals Online (AJOL)

    Hydrolysis and photodegradation of chlorpyrifos a widely used organophosphorus pesticide was studied. Hydrolysis is a dominant ... in aqueous systems. Implication for this study is relevant in understanding the fate of chlorpyrifos once released into the environment with possible application in water/wastewater treatment.

  17. Limited hydrolysis of soybean protein concentrate and isolate with ...

    African Journals Online (AJOL)



    Jul 20, 2009 ... Soy protein concentrate (SPC) and soy protein isolate (SPI) were limited hydrolyzed with trypsin or neutrase under controlled hydrolysis conditions. Eight soybean protein hydrolysates, 4 SPC hydroly- sates and 4 SPI hydrolysates were prepared with degree of hydrolysis (DH) of 1 or 2%. SDS-PAGE was.

  18. Multivariate data analysis of enzyme production for hydrolysis purposes

    DEFF Research Database (Denmark)

    Schmidt, A.S.; Suhr, K.I.


    of the structure in the data - possibly combined with analysis of variance (ANOVA). Partial least squares regression (PLSR) showed a clear connection between the two differentdata matrices (the fermentation variables and the hydrolysis variables). Hence, PLSR was suitable for prediction purposes. The hydrolysis...


    Directory of Open Access Journals (Sweden)



    Full Text Available Every year millions of tons of keratinous wastes are generated worldwide, therefore from an economic and environmental point of view, it is worthwhile developing processes to use and reuse these resources. Many different procedures and methods can be applied to obtain keratin hydrolysates. Microwave heating is based upon the ability of a particular substance to absorb microwave energy and convert the electromagnetic energy to heat (kinetic energy. An extension of this is application in the field of proteomics (the study of proteins and in particular their struc¬ture and function. Large complex proteins can be broken into smaller parts by using microwave heating. This work aimed to perform microwave-assisted hydrolysis of wool keratin wastes in different experimental conditions, and compare it with a conventional method of alkaline hydrolysis, in order to study the efficiency of the treatment induced by this field on the reaction products, subsequently analyzed by specific methods. It was studied the influence of the reaction conditions (reaction time and working recipe on the keratin hydrolysis yield, and also were analysed the structural changes induced by hydrolysis. A significant reduction of reaction time concurrent with a higher hydrolysis yield was obtained by microwave hydrolysis regardless of the alkali mixtures used; better results were obtained for 60 min treatment. IR analysis performed highlight the structural modifications induced by hydrolysis conditions, the process carried out by means of microwaves leading to a lower degradation of hydrolysis products.

  20. Epidemic based modeling of enzymatic hydrolysis of lignocellulosic biomass. (United States)

    Tai, Chao; Arellano, Maria G; Keshwani, Deepak R


    An epidemic based model was developed to describe the enzymatic hydrolysis of a lignocellulosic biomass, dilute sulfuric acid pretreated corn stover. The process of substrate getting adsorbed and digested by enzyme was simulated as susceptibles getting infected by viruses and becoming removed and recovered. This model simplified the dynamic enzyme "infection" process and the catalysis of cellulose into a two-parameter controlled, enzyme behavior guided mechanism. Furthermore, the model incorporates the adsorption block by lignin and inhibition effects on cellulose catalysis. The model satisfactorily predicted the enzyme adsorption and hydrolysis, negative role of lignin, and inhibition effects over hydrolysis for a broad range of substrate and enzyme loadings. Sensitivity analysis was performed to evaluate the incorporation of lignin and other inhibition effects. Our model will be a useful tool for evaluating the effects of parameters during hydrolysis and guide a design strategy for continuous hydrolysis and the associated process control. © 2014 American Institute of Chemical Engineers.

  1. Enzymatic Hydrolysis of Hydrotropic Pulps at Different Substrate Loadings. (United States)

    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.

  2. Trihalomethane hydrolysis in drinking water at elevated temperatures. (United States)

    Zhang, Xiao-Lu; Yang, Hong-Wei; Wang, Xiao-Mao; Karanfil, Tanju; Xie, Yuefeng F


    Hydrolysis could contribute to the loss of trihalomethanes (THMs) in the drinking water at elevated temperatures. This study was aimed at investigating THM hydrolysis pertaining to the storage of hot boiled water in enclosed containers. The water pH value was in the range of 6.1-8.2 and the water temperature was varied from 65 to 95 °C. The effects of halide ions, natural organic matter, and drinking water matrix were investigated. Results showed that the hydrolysis rates declined in the order following CHBrCl2 > CHBr2Cl > CHBr3 > CHCl3. THM hydrolysis was primarily through the alkaline pathway, except for CHCl3 in water at relatively low pH value. The activation energies for the alkaline hydrolysis of CHCl3, CHBrCl2, CHBr2Cl and CHBr3 were 109, 113, 115 and 116 kJ/mol, respectively. No hydrolysis intermediates could accumulate in the water. The natural organic matter, and probably other constituents, in drinking water could substantially decrease THM hydrolysis rates by more than 50%. When a drinking water was at 90 °C or above, the first order rate constants for THM hydrolysis were in the magnitude of 10(-2)‒10(-1) 1/h. When the boiled real tap water was stored in an enclosed container, THMs continued increasing during the first few hours and then kept decreasing later on due to the competition between hydrolysis and further formation. The removal of THMs, especially brominated THMs, by hydrolysis would greatly reduce one's exposure to disinfection by-products by consuming the boiled water stored in enclosed containers. Copyright © 2015 Elsevier Ltd. All rights reserved.

  3. Kinetics of enzymatic hydrolysis of methyl ricinoleate

    Directory of Open Access Journals (Sweden)

    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. Stepwise hydrolysis to improve carbon releasing efficiency from sludge. (United States)

    Liu, Hongbo; Wang, Yuanyuan; Wang, Ling; Yu, Tiantian; Fu, Bo; Liu, He


    Based on thermal alkaline hydrolysis (TAH), a novel strategy of stepwise hydrolysis was developed to improve carbon releasing efficiency from waste activated sludge (WAS). By stepwise increasing hydrolysis intensity, conventional sludge hydrolysis (the control) was divided into four stages for separately recovering sludge carbon sources with different bonding strengths, namely stage 1 (60 °C, pH 6.0-8.0), stage 2 (80 °C, pH 6.0-8.0), stage 3 (80 °C, pH 10.0) and stage 4 (90 °C, pH 12.0). Results indicate stepwise hydrolysis could enhance the amount of released soluble chemical oxygen demand (SCOD) for almost 2 times, from 7200 to 14,693 mg/L, and the released carbon presented better biodegradability, with BOD/COD of 0.47 and volatile fatty acids (VFAs) yield of 0.37 g VFAs/g SCOD via anaerobic fermentation. Moreover, stepwise hydrolysis also improved the dewaterability of hydrolyzed sludge, capillary suction time (CST) reducing from 2500 to 1600 s. Economic assessment indicates stepwise hydrolysis shows less alkali demand and lower thermal energy consumption than those of the control. Furthermore, results of this study help support the concepts of improving carbon recovery in wastewater by manipulating WAS composition and the idea of classifiably recovering the nutrients in WAS. Copyright © 2017 Elsevier Ltd. All rights reserved.

  5. Human milk fat globules: polar lipid composition and in situ structural investigations revealing the heterogeneous distribution of proteins and the lateral segregation of sphingomyelin in the biological membrane. (United States)

    Lopez, Christelle; Ménard, Olivia


    Although human milk fat globules (MFG) are of primary importance since they are the exclusive lipid delivery carriers in the gastrointestinal tract of breast-fed infants, they remain the poorly understood aspect of milk. The objectives of this study were to investigate these unique colloidal assemblies and their interfacial properties, i.e. composition and structure of their biological membrane. In mature breast milk, MFG have a mean diameter of 4-5 microm, a surface area of about 2m(2)/g fat and an apparent zeta potential ζ=-6.7 ± 0.5 mV at 37°C. Human MFG contain 3-4mg polar lipids/g fat as quantified by HPLC/ELSD. The main polar lipids are sphingomyelin (SM; 36-45%, w/w), phosphatidylcholine (19-23%, w/w) and phosphatidylethanolamine (10-15%, w/w). In situ structural investigations of human MFG have been performed using light and confocal microscopy with adapted fluorescent probes, i.e. Nile Red, the extrinsic phospholipid Rh-DOPE, Fast Green and the lectin WGA-488. This study revealed a spatial heterogeneity in the human milk fat globule membrane (MFGM), with the lateral segregation of SM in liquid-ordered phase domains of various shapes and sizes surrounded by a liquid-disordered phase composed of the glycerophospholipids in which the proteins are dispersed. The glycocalyx formed by glycoproteins and cytoplasmic remnents have also been characterised around human MFG. A new model for the structure of the human MFGM is proposed and discussed. The unique composition and lateral organisation of the human MFGM components could be of metabolic significance and have health impact for the infants that need to be further explored. 2010 Elsevier B.V. All rights reserved.

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

    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%).

  7. A possible mechanism underlying the ceramide deficiency in atopic dermatitis: expression of a deacylase enzyme that cleaves the N-acyl linkage of sphingomyelin and glucosylceramide. (United States)

    Imokawa, Genji


    A deficiency of ordinary ceramides in the stratum corneum is an essential etiologic factor for the dry and barrier-disrupted skin of patients with atopic dermatitis (AD). We have proposed that the mechanism underlying that deficiency involves a novel sphingolipid metabolizing enzyme, termed sphingomyelin (SM) glucosylceramide (GCer) deacylase, which hydrolyzes SM or GCer at the acyl site to yield their lysoforms sphingosylphosphorylcholine (SPC) or glucosylsphingosine (GSP) instead of ceramide, leading to the ceramide deficiency in the AD skin. The enzymic characteristics observed showed a pH dependency of catalytic activity with a peak at pH 5.0 and a molecular weight of 40,000. Analytical isoelectric focusing (IEF) chromatography demonstrated that the pI values of SM deacylase, GlcCDase, SMase and ceramidase were 4.2, 7.4, 7.0 and 5.7, respectively. Those enzymic characteristics of SM-GCer deacylase are completely distinct from ceramidase as well as the other known deacylases. Our enzymic measurements demonstrated that SM-GCer deacylase activity is enhanced more than 5-fold in involved stratum corneum, more than 3-fold in uninvolved stratum corneum and approximately 3-fold in the involved epidermis from patients with AD compared with healthy controls. Our findings suggest that the novel enzyme, SM-GCer deacylase, is expressed in situ at significant levels in the epidermis of AD patients. This results in the production of SPC and GSP, instead of ceramides, which leads in turn to the ceramide deficiency seen in the stratum corneum of those patients. It is likely that the biogenesis of SM-GCer deacylase may be critical to the pathogenesis of AD.

  8. Phosphatidylcholine and sphingomyelin profiles vary in Bos taurus indicus and Bos taurus taurus in vitro- and in vivo-produced blastocysts. (United States)

    Sudano, Mateus J; Santos, Vanessa G; Tata, Alessandra; Ferreira, Christina R; Paschoal, Daniela M; Machado, Rui; Buratini, José; Eberlin, Marcos N; Landim-Alvarenga, Fernanda D C


    Lipid droplets, subspecies (Bos taurus indicus vs. Bos taurus taurus), and in vitro culture are known to influence cryopreservation of bovine embryos. Limited information is available regarding differences in membrane lipids in embryo, such as phosphatidylcholines (PC) and sphingomyelins (SM). The objective of the present study was to compare the profiles of several PC and SM species and relate this information to cytoplasmic lipid levels present in Nellore (B. taurus indicus) and Simmental (B. taurus taurus) blastocysts produced in vitro (IVP) or in vivo (ET). Simmental and IVP embryos had more cytoplasmic lipid content than Nellore and ET embryos (n = 30). Blastocysts were submitted to matrix-assisted laser desorption/ionization mass spectrometry. Differences in the PC profile were addressed by principal component analysis. The lipid species with PC (32:1) and PC (34:1) had higher ion abundances in Nellore embryos, whereas PC (34:2) was higher in Simmental embryos. IVP embryos had less abundant ions of PC (32:1), PC (34:2), and PC (36:5) compared to ET embryos. Moreover, ion abundance of PC (32:0) was higher in both Nellore and Simmental IVP embryos compared to ET embryos. Therefore, mass spectrometry profiles of PC and SM species significantly differ with regard to unsaturation level and carbon chain composition in bovine blastocysts due to subspecies and in vitro culture conditions. Because PC abundances of Nellore and Simmental embryos were distinct (34:1 vs. 34:2), as were those of IVP and ET embryos (32:0 vs. 36:5), they are potential markers of postcryopreservation embryonic survival.

  9. Enzymatic hydrolysis of biomass from wood. (United States)

    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. © 2016 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

  10. Catalytic hydrolysis of cellulose into furans (United States)

    Shi, Chengmei; Tao, Furong; Cui, Yuezhi


    Chromium chloride in 4-(3-methylimidazolium-1-yl)butane-1-sulfonic acid hydrogen sulfate (IL-1) was found to effectively catalyze the hydrolysis of microcrystalline cellulose (MCC) at 150°C for 300 min to achieve 87.8% conversion to a slate of products. With a catalytic amount of CrCl3, the yields of 5-hydroxymethyl furfural (HMF) and furfural were up to 32.4 and 15.2%, respectively, small molecules levulinic acid (LA, 10.8%) and the total reducing sugars (TRS, 10.7%) were also generated. Through LC-MSD analysis and mass spectra, dimer of furan compounds as the main by-products were speculated, and the components of gas products were methane, ethane, CO, CO2, and H2. We suggested that IL-1 and CrCl3 exhibited a coordination interaction; the formation of the intermediate via the hydride shift played a key role in the formation of HMF. The catalyst was recycled and exhibited constant activity for five successive trials.

  11. Kinetic Model For Triglyceride Hydrolysis Using Lipase:Review

    Directory of Open Access Journals (Sweden)

    Heri Hermansyah


    Full Text Available Triglyceride hydrolysis using lipase has been proposed as a novel method to produce raw materials in food and cosmetic industries such as diacylglycerol, monoacylglycerol, glycerol and fatty acid. In order to design a reactor for utilizing this reaction on industrial scale, constructing a kinetic model is important. Since the substrates are oil and water, the hydrolysis takes place at oil-water interface. Furthermore, the triglyceride has three ester bonds, so that the hydrolysis stepwise proceeds. Thus, the reaction mechanism is very complicated. The difference between the interfacial and bulk concentrations of the enzyme, substrates and products, and the interfacial enzymatic reaction mechanism should be considered in the model.

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

    DEFF Research Database (Denmark)

    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...

  13. Enzyme loading dependence of cellulose hydrolysis of sugarcane bagasse

    Directory of Open Access Journals (Sweden)

    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.

  14. Energetic approach of biomass hydrolysis in supercritical water. (United States)

    Cantero, Danilo A; Vaquerizo, Luis; Mato, Fidel; Bermejo, M Dolores; Cocero, M José


    Cellulose hydrolysis can be performed in supercritical water with a high selectivity of soluble sugars. The process produces high-pressure steam that can be integrated, from an energy point of view, with the whole biomass treating process. This work investigates the integration of biomass hydrolysis reactors with commercial combined heat and power (CHP) schemes, with special attention to reactor outlet streams. The innovation developed in this work allows adequate energy integration possibilities for heating and compression by using high temperature of the flue gases and direct shaft work from the turbine. The integration of biomass hydrolysis with a CHP process allows the selective conversion of biomass into sugars with low heat requirements. Integrating these two processes, the CHP scheme yield is enhanced around 10% by injecting water in the gas turbine. Furthermore, the hydrolysis reactor can be held at 400°C and 23 MPa using only the gas turbine outlet streams. Copyright © 2014 Elsevier Ltd. All rights reserved.

  15. Enzymatic hydrolysis of pretreated barley and wheat straw

    DEFF Research Database (Denmark)

    Rosgaard, Lisa


    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 indeed potential to boost the enzyme activities in Celluclast (arising from Trichoderma reesei) by addition of small amounts of fermentation broth from fungal sources other than T. reesei at optimal reaction conditions for Celluclast, pH 5, 50 °C. The activity(ies) related to the boosting effect were...... indicated to arise from more efficient or different endoglucanase activities than those found in Celluclast. Evaluating of the extent of hydrolysis using the 4 major enzyme activities in Celluclast, which constituted a complete set of enzymes for hydrolysis of cellulose, showed that the most efficient...

  16. Evaluation of hydrolysis-esterification biodiesel production from wet microalgae. (United States)

    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. Copyright © 2016 Elsevier Ltd. All rights reserved.

  17. Combined wet oxidation and alkaline hydrolysis of polyvinylchloride

    DEFF Research Database (Denmark)

    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...

  18. Effects of processing conditions on hydrolysis of cassava starch ...

    African Journals Online (AJOL)

    amyloglucosidase using 30% initial cassava starch concentration, which produced 152 g/l reducing sugar concentration and DE of 50.9. The total effective operating time was 60 h. Keywords:Cassava starch, hydrolysis, enzyme, dextrose equivalent.

  19. Synthesis of supermacroporous cryogel for bioreactors continuous starch hydrolysis. (United States)

    Guilherme, Ederson Paulo Xavier; de Oliveira, Jocilane Pereira; de Carvalho, Lorendane Millena; Brandi, Igor Viana; Santos, Sérgio Henrique Sousa; de Carvalho, Gleidson Giordano Pinto; Cota, Junio; Mara Aparecida de Carvalho, Bruna


    A bioreactor was built by means of immobilizing alpha-amylase from Aspergillus oryzae by encapsulation, through cryopolymerization of acrylamide monomers for the continuous starch hydrolysis. The starch hydrolysis was evaluated regarding pH, the concentration of immobilized amylase on cryogel, the concentration of starch solution and temperature. The maximum value for starch hydrolysis was achieved at pH 5.0, concentration of immobilized enzyme 111.44 mg amylase /gcryogel , concentration of starch solution 45 g/L and temperature of 35°C. The immobilized enzyme showed a conversion ratio ranging from 68.2 to 97.37%, depending on the pH and temperature employed. Thus, our results suggest that the alpha-amylase from A. oryzae immobilized on cryogel monoliths represents a potential process for industrial production of maltose from starch hydrolysis. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  20. A complete degradation of organophosphates by microwave-assisted hydrolysis

    Czech Academy of Sciences Publication Activity Database

    Jansa, Petr; Čechová, Lucie; Janeba, Zlatko


    Roč. 3, č. 3 (2016), s. 219-226 ISSN 2213-3356 R&D Projects: GA MV VG20102015046 Institutional support: RVO:61388963 Keywords : organophosphates * microwave irradiation * hydrolysis Subject RIV: CC - Organic Chemistry

  1. Hydrolysis of Al3+ from constrained molecular dynamics. (United States)

    Ikeda, Takashi; Hirata, Masaru; Kimura, Takaumi


    We investigated the hydrolysis reactions of Al(3+) in AlCl(3) aqueous solution using the constrained molecular dynamics based on the Car-Parrinello molecular-dynamics method. By employing the proton-aluminum coordination number as a reaction coordinate in the constrained molecular dynamics the deprotonation as well as dehydration processes are successfully realized. From our free-energy difference of DeltaG(0) approximately 8.0 kcal mol(-1) the hydrolysis constant pK(a1) is roughly estimated as 5.8, comparable to the literature value of 5.07. We show that the free-energy difference for the hydrolysis of Al(3+) in acidic conditions is at least 4 kcal mol(-1) higher than that in neutral condition, indicating that the hydrolysis reaction is inhibited by the presence of excess protons located around the hydrated ion, in agreement with the change of the predominant species by pH.

  2. Interaction of Solvents and Mechanical Pretreatment with Enzymatic Lignocellulose Hydrolysis


    Wang, Yumei


    Renewable plant biomass is considered as an alternative raw material for the production of fuels and chemicals instead of decreasing fossil resources. Enzymatic hydrolysis of pretreated lignocellulosic material to produce high sugar concentrations is one important step in a bio-refinery process, and can be operated under moderate conditions without by-products. However, the efficiency of the enzymatic hydrolysis is hindered by lignocellulose recalcitrance. Therefore, pretreatments of the biom...

  3. The human digestive tract has proteases capable of gluten hydrolysis

    Directory of Open Access Journals (Sweden)

    Sergio Gutiérrez


    Conclusion: The digestive tracts of patients with CD and healthy subjects have enzymatic machinery needed for gluten degradation. Patients with CD showed more gluten hydrolysis than did healthy individuals, although, in both cases, a fraction of 33-mer peptide remained intact. Gliadin peptides derived from gastrointestinal digestion, especially the 33-mer, can potentially be used by commensal microbiota from both CD-positive and CD-negative individuals, and differences in bacterial hydrolysis can modify its immunogenic capacity.

  4. Kinetic studies of cellulose enzymatic hydrolysis from pretreated corn cob (United States)

    Stevanie, Jeannie; Kartawiria, Irvan; Abimanyu, Haznan


    Successful utilization of corn cob biomass as raw material in bioethanol production is depending on the hydrolysis process where high level of β-cellulose is converted into glucose. Enzymatic hydrolysis is the common process for this purpose. This study is focusing on the evaluation of hydrolysis of pre-treated corn cob using Novozymes Cellic ® C-Tec2 and H-Tec2 enzymes to obtain the optimum reaction condition and its general reaction kinetics. The corn cob used was pretreated using 10% of NaOH solution. Hydrolysis reactions were conducted in 250 ml Erlenmeyer flask for 72 hour using mixture of C-Tec2 and H-Tec2 enzymes at the fixed ratio of 5:1 and glucose concentration were measured using HPLC. Reaction temperature of 40°C and quantity of 0.5 ml enzyme solution per gram substrate gives the highest reaction rate (0.0123 gram of glucose/gram sample.h) with the glucose yield being 0.089 g glucose/ g substrate. Total conversion of cellulose observed was 11.91 %. Corn cob hydrolysis using C-Tec2 and H-Tec2 enzymes also result in xylose (0.0202 g/g substrate), which can also contribute to bioethanol productivity in further fermentation process. The reaction is following zero order kinetics for the first 8 hours and reaches maximum yield within 10 hours; significantly shorter compared to previous studies of cellulosic material hydrolysis that may take up to 72 hour to complete. Prolonging the hydrolysis of pre-treated corn cob more than 24 hour gives no significant increase in glucose conversion and yield. Hydrolysis temperature range of 40°C to 60°C is in accordance with the manufacturer recommendation for the purpose; however the decrease of reaction rate is observable at temperature 50°C or higher.

  5. Direct injection of superheated steam for continuous hydrolysis reaction

    KAUST Repository

    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.

  6. Hydrolysis of ammonia borane and metal amidoboranes: A comparative study (United States)

    Banu, Tahamida; Debnath, Tanay; Ash, Tamalika; Das, Abhijit K.


    A gas phase mechanistic investigation has been carried out theoretically to explore the hydrolysis pathway of ammonia borane (NH3BH3) and metal amidoboranes (MNH2BH3, M = Li,Na). The Solvation Model based on Density (SMD) has been employed to show the effect of bulk water on the reaction mechanism. Gibbs free energy of solvation has also been computed to evaluate the stabilization of the participating systems in water medium which directly affects the barrier heights in the potential energy surface of hydrolysis reaction. To validate the experimentally observed kinetics studies, we have carried out transition state theory calculations on these hydrolysis reactions. Our result shows that the hydrolysis of both the metal amidoboranes exhibits greatly improved kinetics over the neat NH3BH3 hydrolysis which corroborates well with the experimental observation. Between the two amidoboranes, hydrolysis of LiNH2BH3 is found to be kinetically favored over that of NaNH2BH3, making it a better candidate for releasing molecular hydrogen.

  7. Electron transfer precedes ATP hydrolysis during nitrogenase catalysis (United States)

    Duval, Simon; Danyal, Karamatullah; Shaw, Sudipta; Lytle, Anna K.; Dean, Dennis R.; Hoffman, Brian M.; Antony, Edwin; Seefeldt, Lance C.


    The biological reduction of N2 to NH3 catalyzed by Mo-dependent nitrogenase requires at least eight rounds of a complex cycle of events associated with ATP-driven electron transfer (ET) from the Fe protein to the catalytic MoFe protein, with each ET coupled to the hydrolysis of two ATP molecules. Although steps within this cycle have been studied for decades, the nature of the coupling between ATP hydrolysis and ET, in particular the order of ET and ATP hydrolysis, has been elusive. Here, we have measured first-order rate constants for each key step in the reaction sequence, including direct measurement of the ATP hydrolysis rate constant: kATP = 70 s−1, 25 °C. Comparison of the rate constants establishes that the reaction sequence involves four sequential steps: (i) conformationally gated ET (kET = 140 s−1, 25 °C), (ii) ATP hydrolysis (kATP = 70 s−1, 25 °C), (iii) Phosphate release (kPi = 16 s−1, 25 °C), and (iv) Fe protein dissociation from the MoFe protein (kdiss = 6 s−1, 25 °C). These findings allow completion of the thermodynamic cycle undergone by the Fe protein, showing that the energy of ATP binding and protein–protein association drive ET, with subsequent ATP hydrolysis and Pi release causing dissociation of the complex between the Feox(ADP)2 protein and the reduced MoFe protein. PMID:24062462

  8. Ultrasound enhanced enzymatic hydrolysis of Parthenium hysterophorus: A mechanistic investigation. (United States)

    Singh, Shuchi; Agarwal, Mayank; Bhatt, Aditya; Goyal, Arun; Moholkar, Vijayanand S


    This study has attempted to establish the mechanism of the ultrasound-induced enhancement of enzymatic hydrolysis of pretreated and delignified biomass of Parthenium hysterophorus. A dual approach of statistical optimization of hydrolysis followed by application of sonication at optimum conditions has been adopted. The kinetics of hydrolysis shows a marked 6× increase with sonication, while net sugar yield shows marginal rise of ∼ 20%. The statistical experimental design reveals the hydrolysis process to be enzyme limited. Profile of sugar yield in ultrasound-assisted enzymatic hydrolysis has been analyzed using HCH-1 model coupled with Genetic Algorithm optimization. The trends in the kinetic and physiological parameters of HCH-1 model reveal that sonication enhances enzyme/substrate affinity and reaction velocity of hydrolysis. The product inhibition of enzyme in all forms (free, adsorbed, complexed) also reduces with ultrasound. These effects are attributed to intense micro-convection induced by ultrasound and cavitation in the liquid medium. Copyright © 2015 Elsevier Ltd. All rights reserved.

  9. Development of complete hydrolysis of pectins from apple pomace. (United States)

    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. Copyright © 2014 Elsevier Ltd. All rights reserved.

  10. Impacts of microalgae pre-treatments for improved anaerobic digestion: thermal treatment, thermal hydrolysis, ultrasound and enzymatic hydrolysis. (United States)

    Ometto, Francesco; Quiroga, Gerardo; Pšenička, 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-treatments on three microalgae species, and their impact on the quantity of soluble biomass released in the media and thus on the digestion process yields. The analysis of the composition of the soluble COD released and of the TEM images of the cells showed two main degradation actions associated with the processes: (1) cell wall damage with the release of intracellular AOM (thermal, thermal hydrolysis and ultrasound) and (2) degradation of the cell wall constituents with the release of intracellular AOM and the solubilisation of the cell wall biopolymers (enzymatic hydrolysis). As a result of this, enzymatic hydrolysis showed the greatest biogas yield increments (>270%) followed by thermal hydrolysis (60-100%) and ultrasounds (30-60%). Copyright © 2014 Elsevier Ltd. All rights reserved.

  11. Evidence for the role of lipid rafts and sphingomyelin in Ca2+-gating of Transient Receptor Potential channels in trigeminal sensory neurons and peripheral nerve terminals. (United States)

    Sághy, Éva; Szőke, Éva; Payrits, Maja; Helyes, Zsuzsanna; Börzsei, Rita; Erostyák, János; Jánosi, Tibor Zoltán; Sétáló, György; Szolcsányi, János


    Transient Receptor Potential (TRP) cation channels, such as TRP Vanilloid 1 and TRP Ankyrin repeat domain 1 (TRPV1 and TRPA1) are nocisensors playing important role to signal pain. Two "melastatin" TRP receptors, like TRPM8 and TRPM3 are also expressed in a subgroup of primary sensory neurons. These channels serve as thermosensors with unique thermal sensitivity ranges and are activated also by several exogenous and endogenous chemical ligands inducing conformational changes from various allosteric ("multisteric") sites. We analysed the role of plasma membrane microdomains of lipid rafts on isolated trigeminal (TRG) neurons and TRPV1-expressing CHO cell line by measuring agonist-induced Ca2+ transients with ratiometric technique. Stimulation-evoked calcitonin gene related peptide (CGRP) release from sensory nerve endings of the isolated rat trachea by radioimmunoassay was also measured. Lipid rafts were disrupted by cleaving sphingomyelin (SM) with sphingomyelinase (SMase), cholesterol depletion with methyl β-cyclodextrin (MCD) and ganglioside breakdown with myriocin. It has been revealed that intracellular Ca2+ increase responses evoked by the TRPV1 agonist capsaicin, the TRPA1 agonsits allyl isothiocyanate (AITC) and formaldehyde as well as the TRPM8 activator icilin were inhibited after SMase, MCD and myriocin incubation but the response to the TRPM3 agonist pregnenolon sulphate was not altered. Extracellular SMase treatment did not influence the thapsigargin-evoked Ca2+-release from intracellular stores. Besides the cell bodies, SMase also inhibited capsaicin- or AITC-evoked CGRP release from peripheral sensory nerve terminals, this provides the first evidence for the importance of lipid raft integrity in TRPV1 and TRPA1 gating on capsaicin-sensitive nerve terminals. SM metabolites, ceramide and sphingosine, did not influence TRPA1 and TRPV1 activation on TRG neurons, TRPV1-expressing CHO cell line, and nerve terminals. We suggest, that the hydrophobic

  12. Hydrolysis of aluminum dross material to achieve zero hazardous waste

    Energy Technology Data Exchange (ETDEWEB)

    David, E., E-mail: [National Institute for Research and Development for Cryogenic and Isotopic Technologies, P.O Raureni, P.O. Box 7, 240050 Rm. Valcea (Romania); Kopac, J., E-mail: [University of Ljubljana, Faculty of Mechanical Engineering, Askerceva 6, P.O. Box 394, 1000 Ljubljana (Slovenia)


    Highlights: Black-Right-Pointing-Pointer The hydrolysis of aluminum dross in tap water generates pure hydrogen. Black-Right-Pointing-Pointer Aluminum particles from dross are activated by mechanically milling technique. Black-Right-Pointing-Pointer The process is completely greenhouse gases free and is cleanly to environment. Black-Right-Pointing-Pointer Hydrolysis process leads to recycling of waste aluminum by hydrogen production. - Abstract: A simple method with high efficiency for generating high pure hydrogen by hydrolysis in tap water of highly activated aluminum dross is established. Aluminum dross is activated by mechanically milling to particles of about 45 {mu}m. This leads to removal of surface layer of the aluminum particles and creation of a fresh chemically active metal surface. In contact with water the hydrolysis reaction takes place and hydrogen is released. In this process a Zero Waste concept is achieved because the other product of reaction is aluminum oxide hydroxide (AlOOH), which is nature-friendly and can be used to make high quality refractory or calcium aluminate cement. For comparison we also used pure aluminum powder and alkaline tap water solution (NaOH, KOH) at a ratio similar to that of aluminum dross content. The rates of hydrogen generated in hydrolysis reaction of pure aluminum and aluminum dross have been found to be similar. As a result of the experimental setup, a hydrogen generator was designed and assembled. Hydrogen volume generated by hydrolysis reaction was measured. The experimental results obtained reveal that aluminum dross could be economically recycled by hydrolysis process with achieving zero hazardous aluminum dross waste and hydrogen generation.

  13. Hydrolysis rate constants at 10–25 °C can be more than doubled by a short anaerobic pre-hydrolysis at 35 °C

    NARCIS (Netherlands)

    Zhang, L.; Gao, R.; Naka, A.; Hendrickx, T.L.G.; Rijnaarts, H.H.M.; Zeeman, G.


    Hydrolysis is the first step of the anaerobic digestion of complex wastewater and considered as the rate limiting step especially at low temperature. Low temperature (10–25 °C) hydrolysis was investigated with and without application of a short pre-hydrolysis at 35 °C. Batch experiments were

  14. Starch hydrolysis modeling: application to fuel ethanol production. (United States)

    Murthy, Ganti S; Johnston, David B; Rausch, Kent D; Tumbleson, M E; Singh, Vijay


    Efficiency of the starch hydrolysis in the dry grind corn process is a determining factor for overall conversion of starch to ethanol. A model, based on a molecular approach, was developed to simulate structure and hydrolysis of starch. Starch structure was modeled based on a cluster model of amylopectin. Enzymatic hydrolysis of amylose and amylopectin was modeled using a Monte Carlo simulation method. The model included the effects of process variables such as temperature, pH, enzyme activity and enzyme dose. Pure starches from wet milled waxy and high-amylose corn hybrids and ground yellow dent corn were hydrolyzed to validate the model. Standard deviations in the model predictions for glucose concentration and DE values after saccharification were less than ± 0.15% (w/v) and ± 0.35%, respectively. Correlation coefficients for model predictions and experimental values were 0.60 and 0.91 for liquefaction and 0.84 and 0.71 for saccharification of amylose and amylopectin, respectively. Model predictions for glucose (R2 = 0.69-0.79) and DP4+ (R2 = 0.8-0.68) were more accurate than the maltotriose and maltose for hydrolysis of high-amylose and waxy corn starch. For yellow dent corn, simulation predictions for glucose were accurate (R2 > 0.73) indicating that the model can be used to predict the glucose concentrations during starch hydrolysis.

  15. Enzymatic hydrolysis of fructans in the tequila production process. (United States)

    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.

  16. Ultrasound-assisted lipase catalyzed hydrolysis of aspirin methyl ester. (United States)

    Chiplunkar, Pranali P; Zhao, Xiaoman; Tomke, Prerana D; Noro, Jennifer; Xu, Bo; Wang, Qiang; Silva, Carla; Pratap, Amit P; Cavaco-Paulo, Artur


    The ultrasound-assisted hydrolysis of aspirin methyl ester (AME) was investigated using immobilized Candida antarctica lipase B (CALB) (1%) in the presence of solvents like triolein, chloroform (CHCl3) and dichloromethane (DCM). The effect of ultrasound and the role of water on the conversion rates have also been investigated. Proton nuclear magnetic resonance spectroscopic (1H NMR) was chosen to calculate hydrolysis convertion rates. We observed that lipase-ultrasound assisted hydrolysis of AME in the presence of triolein and water showed the highest hydrolysis conversion rate (65.3%). Herein low water amount played an important role as a nucleophile being crucial for the hydrolysis yields obtained. Lipase activity was affected by the conjugated action of ultrasound and solvents (35.75% of decrease), however not disturbing its hydrolytic efficiency. It was demonstrated that lipase is able to hydrolyse AME to methyl 2-hydroxy benzoate (methyl salicylate), which applications include fragrance agents in food, beverages and cosmetics, or analgesic agent in liniments. Copyright © 2017 Elsevier B.V. All rights reserved.

  17. Hydrolysis of aluminum dross material to achieve zero hazardous waste. (United States)

    David, E; Kopac, J


    A simple method with high efficiency for generating high pure hydrogen by hydrolysis in tap water of highly activated aluminum dross is established. Aluminum dross is activated by mechanically milling to particles of about 45 μm. This leads to removal of surface layer of the aluminum particles and creation of a fresh chemically active metal surface. In contact with water the hydrolysis reaction takes place and hydrogen is released. In this process a Zero Waste concept is achieved because the other product of reaction is aluminum oxide hydroxide (AlOOH), which is nature-friendly and can be used to make high quality refractory or calcium aluminate cement. For comparison we also used pure aluminum powder and alkaline tap water solution (NaOH, KOH) at a ratio similar to that of aluminum dross content. The rates of hydrogen generated in hydrolysis reaction of pure aluminum and aluminum dross have been found to be similar. As a result of the experimental setup, a hydrogen generator was designed and assembled. Hydrogen volume generated by hydrolysis reaction was measured. The experimental results obtained reveal that aluminum dross could be economically recycled by hydrolysis process with achieving zero hazardous aluminum dross waste and hydrogen generation. Copyright © 2012 Elsevier B.V. All rights reserved.

  18. Granular starch hydrolysis for fuel ethanol production (United States)

    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.


    Directory of Open Access Journals (Sweden)

    Leyanis Rodríguez Rodríguez


    Full Text Available The main objective of this work was the production of syrups dextrinized by enzymatic hydrolysis of starch red sorghum CIAPR-132 using α-amylase on solutions at different concentrations, with different concentrations of enzyme and enzyme hydrolysis time. The response variable was the dextrose equivalent in each obtained syrup (ED using the modified Lane-Eynon method. In some of the experiments, we used a full factorial design 23 and in others we worked with intermediate concentration and higher hydrolysis time with different levels of enzyme. The obtained products were syrups dextrinized ED between 10,25 and 33,97% (values we can find within the established ones for these types of syrups, which can be used for their functional properties as intermediates syrups or as raw material for different processes of the food industry. This allows you to set a pattern for the use of sorghum feedstock in unconventional obtaining products from its starch.

  20. Hydrolysis of Toxic Natural Glucosides Catalyzed by Cyclodextrin Dicyanohydrins

    DEFF Research Database (Denmark)

    Bjerre, Jeannette; Nielsen, Erik Holm; Bols, Mikael


    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 impre......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...... degree of catalysis was also found for the toxic hydroxycoumarin esculin. A novel synthesized diaminomethyl ß-cyclodextrin showed a weak catalysis of p-nitrophenyl ß-D-glucopyranoside hydrolysis....

  1. Enzyme hydrolysis kinetics of micro-grinded maize straws. (United States)

    Hu, Jianjun; Jing, Yanyan; Zhang, Quanguo; Guo, Jie; Lee, Duu-Jong


    This study applied micro-grinding to disintegrate the maize straws and then use the micro-grinded straws of particle sizes particle size 53-61, 80-96 or 150-180μm, for subsequent enzyme hydrolysis tests. The reducing sugar productivity was increased with reducing particle size. A kinetic model considering product inhibition was developed as follows t=aln[S]0[S]0-[P]+b[P], where S, P and t are the substrate, enzyme and hydrolysis time, respectively, and a and b are fitting parameters. The initial substrate concentration is proportional to the total exposed surface area. Additionally, the mechanical grinding can increase the biomass affinity for enzyme attack, suggesting the enhanced local action of shearing on the fiber matrix surfaces. The enhanced hydrolysis efficiency of the micro-grinded straws is welcomed by the subsequent refinery steps. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. Mechanistic kinetic models of enzymatic cellulose hydrolysis-A review. (United States)

    Jeoh, Tina; Cardona, Maria J; Karuna, Nardrapee; Mudinoor, Akshata R; Nill, Jennifer


    Bioconversion of lignocellulose forms the basis for renewable, advanced biofuels, and bioproducts. Mechanisms of hydrolysis of cellulose by cellulases have been actively studied for nearly 70 years with significant gains in understanding of the cellulolytic enzymes. Yet, a full mechanistic understanding of the hydrolysis reaction has been elusive. We present a review to highlight new insights gained since the most recent comprehensive review of cellulose hydrolysis kinetic models by Bansal et al. (2009) Biotechnol Adv 27:833-848. Recent models have taken a two-pronged approach to tackle the challenge of modeling the complex heterogeneous reaction-an enzyme-centric modeling approach centered on the molecularity of the cellulase-cellulose interactions to examine rate limiting elementary steps and a substrate-centric modeling approach aimed at capturing the limiting property of the insoluble cellulose substrate. Collectively, modeling results suggest that at the molecular-scale, how rapidly cellulases can bind productively (complexation) and release from cellulose (decomplexation) is limiting, while the overall hydrolysis rate is largely insensitive to the catalytic rate constant. The surface area of the insoluble substrate and the degrees of polymerization of the cellulose molecules in the reaction both limit initial hydrolysis rates only. Neither enzyme-centric models nor substrate-centric models can consistently capture hydrolysis time course at extended reaction times. Thus, questions of the true reaction limiting factors at extended reaction times and the role of complexation and decomplexation in rate limitation remain unresolved. Biotechnol. Bioeng. 2017;114: 1369-1385. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

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

    DEFF Research Database (Denmark)

    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...

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

    DEFF Research Database (Denmark)

    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...

  5. Visualizing phosphodiester-bond hydrolysis by an endonuclease

    DEFF Research Database (Denmark)

    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....... This 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...

  6. Single Molecule Study of Cellulase Hydrolysis of Crystalline Cellulose

    Energy Technology Data Exchange (ETDEWEB)

    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.

  7. Proximity effect on the general base catalysed hydrolysis of amide ...

    Indian Academy of Sciences (India)

    A constant value of absorbance could not be achieved even after 3 days. However, independent measurements. (by mixing the corresponding amines, salicylic acid and. NaOH, I = 0.5 mol dm. −3) showed that a reaction mix- ture for the complete hydrolysis of the amides (I–IV) to the corresponding amines and salicylate had ...

  8. Hydrolysis of raw tuber starches by amylase of Aspergillus niger ...

    African Journals Online (AJOL)

    Hydrolysis of raw tuber starches by amylase of Aspergillus niger AM07 isolated from the soil. ... The crude amylase preparation of A. niger AM07 had temperature and pH optima activities at 60oC and 4.0 respectively. The optimum substrate concentration was 3 %. The action of the crude amylase of A. niger on raw tuber ...

  9. Small peptides hydrolysis in dry-cured meats. (United States)

    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. Copyright © 2015 Elsevier B.V. All rights reserved.

  10. Micellar Catalysis Of The Hydrolysis Of Aryltrifluoroacetates | Ayinde ...

    African Journals Online (AJOL)

    The neutral hydrolysis of aryl trifluoroacetates was investigated in micellar media of cetyl trimethylammonium bromide (CTAB) and sodium dodecylsulphate (SDS). The reaction is catalyzed by both CTAB and SDS micelles, with larger rate enhancement in CTAB. The reaction mechanism (B AC 3) is the same in both micellar ...

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

    DEFF Research Database (Denmark)

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


    .7% of the theoretical maximum value. Pretreatment at 200 C with oxygen exhibited enhanced enzymatic efficiency but lower xylose recovery and formation of the degradation products such as acetate, furfural and HMF of 7.6, 3.3 and 1.0 g/L, respectively. In the hydrolysis, the total sugars (glucose + xylose) yielded...

  12. modelling and simulation of the batch hydrolysis of acetic ing

    African Journals Online (AJOL)


    acid has been studied with a number of techniques for measuring reaction rate reported in literature. Ampelli et al, [7] integrated an ultraviolet-visible. (UV-Vis) spectrometer with an isoperibolic reaction calorimeter to study the kinetics of hydrolysis of acetic anhydride. An acid–base indicator was added to the reaction mixture ...

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

    NARCIS (Netherlands)

    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

  14. Penicillin Hydrolysis: A Kinetic Study of a Multistep, Multiproduct Reaction. (United States)

    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)

  15. Modelling and Simulation of the Batch Hydrolysis of Acetic ...

    African Journals Online (AJOL)

    The kinetic modelling of the batch synthesis of acetic acid from acetic anhydride was investigated. The kinetic data of the reaction was obtained by conducting the hydrolysis reaction in a batch reactor. A dynamic model was formulated for this process and simulation was carried out using gPROMS® an advanced process ...

  16. Eggshells – assisted hydrolysis of banana pulp for biogas production

    African Journals Online (AJOL)


    The least biogas yield of 10 mL was obtained in digester. C5 with 9 g of calcined eggshells additive. Key words: Anaerobic digestion, banana pulp hydrolysis biogas, eggshells. INTRODUCTION. The energy and global warming crisis, stimulates the need for development of renewable energy worldwide. (Buasri et al., 2013).

  17. Eggshells – assisted hydrolysis of banana pulp for biogas production

    African Journals Online (AJOL)

    In this study, pretreatment of banana pulp using eggshells in both calcined and un-calcined forms to examine the extent of hydrolysis was conducted. Reactor CO containing banana pulp and inoculum but with no eggshells added was used as the control, while reactors C1, C2, C3, C4, and C5 containing banana pulp and ...

  18. The trypsin-catalyzed hydrolysis of monomolecular films of lysylphosphatidylglycerol

    NARCIS (Netherlands)

    Gould, R.M.; Dawson, R.M.C.


    The hydrolysis by trypsin of the bacterial phospholipid, lysylphosphatidyl-glycerol has been studied at the air-water interface. High specific activity [14C]-lysylphosphatidylglycerol was prepared biosynthetically and the trypsin action followed by measuring the loss of surface radioactivity from a

  19. Optimisation of Dilute Sulphuric Acid Hydrolysis of Waste ...

    African Journals Online (AJOL)

    A statistical model was developed for the optimisation of the process variables using response surface methodology (RSM). The optimal hydrolysis conditions that resulted in the maximum total reducing sugar concentration were: acid concentration, 1.49 %w/w; temperature, 200°C; time, 20 min and liquid to solid ratio, ...

  20. Hydrolysis of proteinaceous tannery solid waste for the production of ...

    African Journals Online (AJOL)

    Hydrolysis of proteinaceous tannery solid waste for the production of extracellular acidic protease by Selenomonas ruminantium. ... The enzyme had good stability in the presence of non-ionic surfactants like tween 20, tween 40, tween 80 and triton X100 and also in the presence of solvents like methanol, ethanol and ...

  1. Plant dried powders as biocatalysts: Hydrolysis of 1- phenylpropanol ...

    African Journals Online (AJOL)

    The hydrolytic ability of plant dried powders, lyophilized or acetone dried, was tested on the hydrolysis of racemic 1-phenylpropanol acetate. Most of the twenty powders tested showed hydrolytic activity, however the best values of conversion and enantioselectivity were reached with the lyophilized powder of nopal (27% ...

  2. A Comparative Kinetic Study of Acidic Hydrolysis of Wastes ...

    African Journals Online (AJOL)

    Michael Horsfall

    of acid hydrolysis of cellulose isolated from banana skin, cowpea shells, maize stalks and rice husk (agricultural waste) were studied at temperature ranging ... Many of these agro-wastes are allowed to rot away not utilized (Obot et al., 2008). ..... sustainable supply of liquid fuel in china. Biotechnol J. 1:1235. Rahman, S.H.A. ...

  3. Optimization of peptide production by enzymatic hydrolysis of tuna ...

    African Journals Online (AJOL)

    Optimization of peptide production by enzymatic hydrolysis of tuna dark muscle by-product using commercial proteases. ... The PDF file you selected should load here if your Web browser has a PDF reader plug-in installed (for example, a recent version of Adobe Acrobat Reader). If you would like more information about ...

  4. Hydrolysis of maize starch using amylolytic enzymes extracted from ...

    African Journals Online (AJOL)

    Amylases, a-amylase (EC, α-1, 4-glucan-4-glucanohydrolase) and glucoamylase (EC, α-1, 4; α-1, 6-glucan glucohydrolase; amyloglucosidase), extracted and partially purified from sorghum malt were used to hydrolyze maize starch. The process and products of the enzymatic hydrolysis were also compared ...

  5. Maltodextrin hydrolysis with glucoamylase immobilized in polyacrylamide gel

    Energy Technology Data Exchange (ETDEWEB)

    Yankov, D.; Peeva, L.; Beschkov, V. (Inst. of Chemical Engineering, Bulgarian Academy of Sciences, Sofia (Bulgaria))


    The immobilization of glucoamylase by entrapping in a polyacrylamide gel has been studied. The optimum values of pH and temperature for the immobilized enzyme have been determinated. The influence of worked off cycles, substrate concentration and initial degree of hydrolysis on the final conversion of starch have been investigated. (orig.).

  6. Effect of acid and alkaline hydrolysis on the concentrations of ...

    African Journals Online (AJOL)

    Thevetia peruviana seeds cake were defatted and then treated with varying concentrations each of hydrochloric acid, sodium hydroxide and calcium hydroxide solutions. Each product of hydrolysis was extracted with chloroform to isolate aglycones, the toxins of the seed. Various concentrations of hydrochloric acid and ...

  7. Effects of hydrolysis and carbonization reactions on hydrochar production. (United States)

    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. Copyright © 2015 Elsevier Ltd. All rights reserved.

  8. Thermal hydrolysis for sewage treatment: A critical review. (United States)

    Barber, W P F


    A review concerning the development and applicability of sewage sludge thermal hydrolysis especially prior to anaerobic digestion is presented. Thermal hydrolysis has proven to be a successful approach to making sewage sludge more amenable to anaerobic digestion. Currently there are 75 facilities either in operation or planning, spanning several continents with the first installation in 1995. The reported benefits of thermal hydrolysis relate to: increased digestion loading rate due to altered rheological properties, improved biodegradation of (especially activated) sludge and enhanced dewaterability. In spite of its relative maturity, there has been no attempt to perform a critical review of the pertinent literature relating to the technology. Closer look at the literature reveals complications with comparing both experimental- and full-scale results due to differences in experimental set-up and capability, and also site-specific conditions at full-scale. Furthermore, it appears that understanding of thermodynamic and rheological properties of sludge is key to optimizing the process, however these parameters are largely overlooked by the literature. This paper aims to bridge these complexities in order to elucidate the benefits of thermal hydrolysis for sewage treatment, and makes recommendations for further development and research. Copyright © 2016 Elsevier Ltd. All rights reserved.

  9. The Preparation and Enzymatic Hydrolysis of a Library of Esters (United States)

    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…

  10. Super acid catalysed sequential hydrolysis/cycloisomerization of o ...

    Indian Academy of Sciences (India)

    Abstract. Synthesis of isocoumarins and related compounds via triflic acid promoted hydrolysis/cyclization sequence of 2-(alkynyl)benzamides under microwave condition was achieved. The substrate scope of the reac- tion was broad to include not only aromatic but also polyaromatic and heteroaromatic motifs, thus ...

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

    NARCIS (Netherlands)

    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.

  12. Hydrolysis conditions for the analysis for sulphur amino acids in ...

    African Journals Online (AJOL)

    A study of methods of analysis for sulphur amino acids in feeds was undertaken with a view to finding methods suitable for the routine analysis of feedstuffs. In the case of cyst(e)ine, it was found that the results ob- tained from simple oxidation and hydrolysis with a mixture of dimethylsulphoxide (OMSO) and hydrochloric ...

  13. Validation of lignocellulosic biomass carbohydrates determination via acid hydrolysis. (United States)

    Zhou, Shengfei; Runge, Troy M


    This work studied the two-step acid hydrolysis for determining carbohydrates in lignocellulosic biomass. Estimation of sugar loss based on acid hydrolyzed sugar standards or analysis of sugar derivatives was investigated. Four model substrates (starch, holocellulose, filter paper and cotton) and three levels of acid/material ratios (7.8, 10.3 and 15.4, v/w) were studied to demonstrate the range of test artifacts. The method for carbohydrates estimation based on acid hydrolyzed sugar standards having the most satisfactory carbohydrate recovery and relative standard deviation. Raw material and the acid/material ratio both had significant effect on carbohydrate hydrolysis, suggesting the acid to have impacts beyond a catalyst in the hydrolysis. Following optimal procedures, we were able to reach a carbohydrate recovery of 96% with a relative standard deviation less than 3%. The carbohydrates recovery lower than 100% was likely due to the incomplete hydrolysis of substrates, which was supported by scanning electron microscope (SEM) images. Copyright © 2014 Elsevier Ltd. All rights reserved.

  14. hippurate hydrolysis and christie, atkins, munch-peterson tests as ...

    African Journals Online (AJOL)



    Jan 1, 2000 ... Objective: To evaluate the Christie, Atkins, Munch-Peterson (CAMP) and hippurate hydrolysis reactions as diagnostic tools for Streptococcus agalactiae carriage in pregnancy. Design: Observational, analytical case-control study. Setting: Hospital-based study in a primary and a tertiary health care institution.

  15. Heppurate hydrolysis and Christie, Atkins, Munch-Peterson tests ...

    African Journals Online (AJOL)

    Objectives: To evaluate the Christie, Atkins, Munch-Peterson (CAMP) and hippurate hydrolysis reactions as diagnostic tools for Streptococcus agalactiae carriage in pregnancy. Design: Observational, analytical case-control study. Setting: Hospital-based study in a primary and a tertiary health care institution. Patients: One ...

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

    Energy Technology Data Exchange (ETDEWEB)

    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.

  17. Bioethanol production: Pretreatment and enzymatic hydrolysis of softwood

    Energy Technology Data Exchange (ETDEWEB)

    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.

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

    NARCIS (Netherlands)

    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

  19. Accelerated hydrolysis of substituted cellulose for potential biofuel production: kinetic study and modeling. (United States)

    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. Copyright © 2015 Elsevier Ltd. All rights reserved.

  20. 40 CFR 721.10152 - Oxirane, substituted silylmethyl-, hydrolysis products with alkanol zirconium(4+) salt and silica... (United States)


    ...-, hydrolysis products with alkanol zirconium(4+) salt and silica, acetates (generic). 721.10152 Section 721... Oxirane, substituted silylmethyl-, hydrolysis products with alkanol zirconium(4+) salt and silica... substance identified generically as oxirane, substituted silylmethyl-, hydrolysis products with alkanol...

  1. A Dynamic Model for Cellulosic Biomass Hydrolysis: a Comprehensive Analysis and Validation of Hydrolysis and Product Inhibition Mechanisms

    DEFF Research Database (Denmark)

    Tsai, Chien Tai; Morales Rodriguez, Ricardo; Sin, Gürkan


    The objective of this study is to perform a comprehensive enzyme kinetics analysis in view of validating and consolidating a semimechanistic kinetic model consisting of homogeneous and heterogeneous reactions for enzymatic hydrolysis of lignocellulosic biomass proposed by the U.S. National Renewa...

  2. Effect of acid hydrolysis of Garcinia kola (bitter kola) pulp waste on ...

    African Journals Online (AJOL)

    Effect of acid hydrolysis of bitter kola (Garcinia kola) pulp wastes on the production of CM-cellulase and -glucosidase using Aspergillus niger was investigated. Hydrolysis for 3 h with 2.5 M sulphuric acid yielded highest fermentable sugar. Acid hydrolysis enhanced CM-cellulase and -glucosidase levels by 500% at 96 h and ...

  3. Stimulation and inhibition of enzymatic hydrolysis by organosolv lignins as determined by zeta potential and hydrophobicity (United States)

    Yang Huang; Shaolong Sun; Chen Huang; Qiang Yong; Thomas Elder; Maobing Tu


    Background: Lignin typically inhibits enzymatic hydrolysis of cellulosic biomass, but certain organosolv lignins or lignosulfonates enhance enzymatic hydrolysis. The hydrophobic and electrostatic interactions between lignin and cellulases play critical roles in the enzymatic hydrolysis process. However, how to incorporate these two...

  4. Bioethanol production: an integrated process of low substrate loading hydrolysis-high sugars liquid fermentation and solid state fermentation of enzymatic hydrolysis residue. (United States)

    Chu, Qiulu; Li, Xin; Ma, Bin; Xu, Yong; Ouyang, Jia; Zhu, Junjun; Yu, Shiyuan; Yong, Qiang


    An integrated process of enzymatic hydrolysis and fermentation was investigated for high ethanol production. The combination of enzymatic hydrolysis at low substrate loading, liquid fermentation of high sugars concentration and solid state fermentation of enzymatic hydrolysis residue was beneficial for conversion of steam explosion pretreated corn stover to ethanol. The results suggested that low substrate loading hydrolysis caused a high enzymatic hydrolysis yield; the liquid fermentation of about 200g/L glucose by Saccharomyces cerevisiae provided a high ethanol concentration which could significantly decrease cost of the subsequent ethanol distillation. A solid state fermentation of enzymatic hydrolysis residue was combined, which was available to enhance ethanol production and cellulose-to-ethanol conversion. The results of solid state fermentation demonstrated that the solid state fermentation process accompanied by simultaneous saccharification and fermentation. Copyright © 2012 Elsevier Ltd. All rights reserved.

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

    Energy Technology Data Exchange (ETDEWEB)

    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

  6. Modeling the yield of hydrolysis process of leather shavings

    Directory of Open Access Journals (Sweden)

    Vaskova Hana


    Full Text Available In this paper the mathematical modeling of collagen hydrolyzate yield from shavings, a leather waste material, is dealt. Besides its function, leather industry is perceived as a consumer of resources and producer of pollutants. The process of alkaline enzymatic hydrolysis offers a reasonable method of processing part of solid waste arising from leather industry with the aim of further utilization of collagenous share. The model is based on the total mass and dry matter balances of technology and its individual components to optimize the hydrolysate yields. The results are visualised in 3D. The experimental laboratory data of alkaline enzymatic hydrolysis are used for comparison with the model and are discussed.

  7. Effect of Limited Hydrolysis on Traditional Soy Protein Concentrate

    Directory of Open Access Journals (Sweden)

    Mirjana B. Pesic


    Full Text Available The influence of limited proteolysis of soy protein concentrate on proteinextractability, the composition of the extractable proteins, their emulsifying properties andsome nutritional properties were investigated. Traditional concentrate (alcohol leachedconcentrate was hydrolyzed using trypsin and pepsin as hydrolytic agents. Significantdifferences in extractable protein composition between traditional concentrate and theirhydrolysates were observed by polyacrylamide gel electrophoresis (PAGE and by SDSPAGE.All hydrolysates showed better extractability than the original protein concentrate,whereas significantly better emulsifying properties were noticed at modified concentratesobtained by trypsin induced hydrolysis. These improved properties are the result of twosimultaneous processes, dissociation and degradation of insoluble alcohol-induced proteinaggregates. Enzyme induced hydrolysis had no influence on trypsin-inibitor activity, andsignificantly reduced phytic acid content.

  8. Theoretical study on alkaline hydrolysis of trinitrotoluene: later steps

    Directory of Open Access Journals (Sweden)

    Liudmyla K. Sviatenko


    Full Text Available Alkaline hydrolysis is an effective method to destroy such the pollutant as 2,4,6-trinitrotoluene (TNT in solution and in well-mixed soil. The mechanism of hydrolytic transformation of polynegative complex, which is one of the products of early stages of TNT hydrolysis, was theoretically investigated at the SMD(Pauling/M06-2X/6-31+G(d,p level under alkali condition. The studied process consists of more than twenty steps and includes a six-membered cycle cleavage and sequenced [1,3]-hydrogen migration and C-C bond rupture. The highest energy barrier is observed for interaction of nitromethanide with hydroxide. The most exothermic steps are C–C bonds breaking. As a result final products such as formate, acetate, ammonium, and nitrogen are formed.

  9. Acid hydrolysis of sugar beet pulp as pretreatment for fermentation

    Energy Technology Data Exchange (ETDEWEB)

    Chamy, R.; Illanes, A.; Aroca, G.; Nunez, L. [Universidad Catolica de Valparaiso (Chile). School of Biochemical Engineering


    The purpose of this work is to optimize substrate pretreatment by selectively solubilizing the hemicellulose fraction to render a cellulose-enriched fraction for further fermentation or enzyme hydrolysis. Hemicellulose hydrolyzate, usually a waste stream, is proposed to be fermented by the pentose-utilizing yeast Pichia stipitis. Sugar beet pulp (SBP) was chosen as a substance due to its low lignin content which makes substance pretreatment simpler and less expensive. Hemicellulose utilization is very important in the case of SBP, whose hemicellulose content is as high as 50%. The best conditions for SBP pretreatment were: 55 g/l of 32-50 mesh SBP, 1.1 g H{sub 2}SO{sub 4}/g SBP, 90 min at 80{sup o}C and 400 rpm. Under such conditions, 86.3% and 7.8% of cellulose and hemicellulose hydrolysis, respectively, were obtained. (author)


    Directory of Open Access Journals (Sweden)

    Merçon F.


    Full Text Available This work deals with enzymatic hydrolysis of babassu oil by immobilized lipase in membrane reactors of two types: a flat plate nylon membrane and a hollow fiber polyetherimide membrane on which surface commercial lipases were immobilized by adsorption. Experiments conducted in the hollow fiber reactor showed that during the immobilization step enzyme adsorption followed a sigmoid model, with a maximum adsorption equilibrium time of 30 minutes. Concerning the hydrodynamics of the liquid phases, the results indicate that main diffusional limitations occurred in the organic phase. The amount of protein immobilized and the maximum productivity were, respectively, 1.97 g/m2 and 44 m molH+/m2.s for the hollow fiber and 1.2 g/m2 and 56 m molH+/m2.s for the flat and plate membrane. Both reactors were able to perform the hydrolysis reaction, while maintaining absolute separation of the two phases by the membrane

  11. A DFT investigation of methanolysis and hydrolysis of triacetin

    CERN Document Server

    Limpanuparb, Taweetham; Tantirungrotechai, Yuthana; 10.1016/j.theochem.2010.05.022


    The thermodynamic and kinetic aspects of the methanolysis and hydrolysis reactions of glycerol triacetate or triacetin, a model triacylglycerol compound, were investigated by using Density Functional Theory (DFT) at the B3LYP/6-31++G(d,p) level of calculation. Twelve elementary steps of triacetin methanolysis were studied under acid-catalyzed and base-catalyzed conditions. The mechanism of acid-catalyzed methanolysis reaction which has not been reported yet for any esters was proposed. The effects of substitution, methanolysis/hydrolysis position, solvent and face of nucleophilic attack on the free energy of reaction and activation energy were examined. The prediction confirmed the facile position at the middle position of glycerol observed by NMR techniques. The calculated activation energy and the trends of those factors agree with existing experimental observations in biodiesel production.

  12. Computational redesign of a mononuclear zinc metalloenzyme for organophosphate hydrolysis

    DEFF Research Database (Denmark)

    Khare, Sagar D.; Kipnis, Yakov; Greisen, Per Junior


    . The designed enzyme efficiently catalyzes the hydrolysis of the R(P) isomer of a coumarinyl analog of the nerve agent cyclosarin, and it shows marked substrate selectivity for coumarinyl leaving groups. Computational redesign of native enzyme active sites complements directed evolution methods and offers......The ability to redesign enzymes to catalyze noncognate chemical transformations would have wide-ranging applications. We developed a computational method for repurposing the reactivity of metalloenzyme active site functional groups to catalyze new reactions. Using this method, we engineered a zinc......-containing mouse adenosine deaminase to catalyze the hydrolysis of a model organophosphate with a catalytic efficiency (k(cat)/K(m)) of ~10(4) M(-1) s(-1) after directed evolution. In the high-resolution crystal structure of the enzyme, all but one of the designed residues adopt the designed conformation...

  13. Simultaneous hydrolysis-esterification of wet microalgal lipid using acid. (United States)

    Takisawa, Kenji; Kanemoto, Kazuyo; Kartikawati, Muliasari; Kitamura, Yutaka


    This research demonstrated hydrolysis of wet microalgal lipid and esterification of free fatty acid (FFA) using acid in one-step process. The investigation of simultaneous hydrolysis-esterification (SHE) of wet microalgal lipid was conducted by using L27 orthogonal design and the effects of water content, volume of sulphuric acid, volume of methanol, temperature and time on SHE were examined. As a result, water content was found to be the most effective factor. The effects of various parameters on fatty acid methyl ester (FAME) content and equilibrium relation between FAME and FFA were also examined under water content 80%. Equimolar amounts of sulphuric acid and hydrochloric acid showed similar results. This method has great potential in terms of biodiesel production from microalgae since no organic solvents are used. Copyright © 2013 Elsevier Ltd. All rights reserved.

  14. Kinetics of the Acid Hydrolysis of Potassium Trans-dioxalatobis ...

    African Journals Online (AJOL)

    Kinetics of the Acid Hydrolysis of Potassium Trans-dioxalatobis(Ethylnicotinate) Chromium(iii). T.B.R. Yormah, D.V.A. Fode, M.K. Kormoh. Abstract. The kinetics of the aquation of trans-Cr(ox)2(py.3COOEt)2 - has been studied spectrophotometrically in aqueous HClO4-NaClO4 media. The results indicate a first-order ...

  15. Synthesis of zirconia colloidal dispersions by forced hydrolysis

    Directory of Open Access Journals (Sweden)



    Full Text Available Different zirconia colloidal dispersions (sols were prepared from zirconyl oxynitrate and zirconyl oxychloride solutions by forced hydrolysis. Vigorously stirred acidic solutions of these salts were refluxed at 102 oC for 24 h. Characterization of the obtained sols (pH, solid phase content, crystal structure was performed by potentiometric, XRD, TGA/DTA and SEM measurements. The prepared sols contained almost spherical monoclinic hydrated zirconia particles 7–10 nm in diameter.

  16. Efficient Phagocytosis Requires Triacylglycerol Hydrolysis by Adipose Triglyceride Lipase*


    Chandak, Prakash G.; Radovi?, Branislav; Aflaki, Elma; Kolb, Dagmar; Buchebner, Marlene; Fr?hlich, Eleonore; Magnes, Christoph; Sinner, Frank; Haemmerle, Guenter; Zechner, Rudolf; Tabas, Ira; Levak-Frank, Sanja; Kratky, Dagmar


    Macrophage phagocytosis is an essential biological process in host defense and requires large amounts of energy. To date, glucose is believed to represent the prime substrate for ATP production in macrophages. To investigate the relative contribution of free fatty acids (FFAs) in this process, we determined the phagocytosis rates in normal mouse macrophages and macrophages of adipose triglyceride lipase (ATGL)-deficient mice. ATGL was shown to be the rate-limiting enzyme for the hydrolysis of...

  17. Modeling the yield of hydrolysis process of leather shavings


    Vaskova Hana; Kocurek Pavel


    In this paper the mathematical modeling of collagen hydrolyzate yield from shavings, a leather waste material, is dealt. Besides its function, leather industry is perceived as a consumer of resources and producer of pollutants. The process of alkaline enzymatic hydrolysis offers a reasonable method of processing part of solid waste arising from leather industry with the aim of further utilization of collagenous share. The model is based on the total mass and dry matter balances of technology ...

  18. Sugar Production from Durian Peel by Acid Hydrolysis

    African Journals Online (AJOL)

    Valued Acer Customer


    Aug 14, 2013 ... Agricultural waste is mainly composed of cellulose and hemicelluloses which can be converted to sugars. The inexpensive sugar from durian peel was hydrolyzed with H2SO4, HCl and H3PO4 at a concentration of 0.5 to 2.0% (v/v). The hydrolysis times ranging from 15 to 60 min were investigated when the ...

  19. Investigations Concerning Hydrolysis and Stabilization of Antiradiation Compounds (United States)


    The present Waters RCM-100 separation column was originally chosen from a number of different reverse phase columns based on superior performance 1n...the development of the assay were the instability of WR 2721 in acidic media and its lack of a suitable chromophore. A pH- hydrolysis rate profile...Eiaiiy M. Gause Southwest Foundation for Research and Educacion San Anconlo, Texas 78284 Lawrence Fleclcensteln Division of Experimental Therapeutics

  20. Endo-exo Synergism in Cellulose Hydrolysis Revisited* (United States)

    Jalak, Jürgen; Kurašin, Mihhail; Teugjas, Hele; Väljamäe, Priit


    Synergistic cooperation of different enzymes is a prerequisite for efficient degradation of cellulose. The conventional mechanistic interpretation of the synergism between randomly acting endoglucanases (EGs) and chain end-specific processive cellobiohydrolases (CBHs) is that EG-generated new chain ends on cellulose surface serve as starting points for CBHs. Here we studied the hydrolysis of bacterial cellulose (BC) by CBH TrCel7A and EG TrCel5A from Trichoderma reesei under both single-turnover and “steady state” conditions. Unaccountable by conventional interpretation, the presence of EG increased the rate constant of TrCel7A-catalyzed hydrolysis of BC in steady state. At optimal enzyme/substrate ratios, the “steady state” rate of synergistic hydrolysis became limited by the velocity of processive movement of TrCel7A on BC. A processivity value of 66 ± 7 cellobiose units measured for TrCel7A on 14C-labeled BC was close to the leveling off degree of polymerization of BC, suggesting that TrCel7A cannot pass through the amorphous regions on BC and stalls. We propose a mechanism of endo-exo synergism whereby the degradation of amorphous regions by EG avoids the stalling of TrCel7A and leads to its accelerated recruitment. Hydrolysis of pretreated wheat straw suggested that this mechanism of synergism is operative also in the degradation of lignocellulose. Although both mechanisms of synergism are used in parallel, the contribution of conventional mechanism is significant only at high enzyme/substrate ratios. PMID:22733813

  1. Enzymatic hydrolysis of defatted mackerel protein with low bitter taste (United States)

    Hou, Hu; Li, Bafang; Zhao, Xue


    Ultrasound-assisted solvent extraction was confirmed as a novel, effective method for separating lipid from mackerel protein, resulting in a degreasing rate (DR) of 95% and a nitrogen recovery (NR) of 88.6%. To obtain protein hydrolysates with high nitrogen recovery and low bitter taste, enzymatic hydrolysis was performed using eight commercially available proteases. It turned out that the optimum enzyme was the `Mixed enzymes for animal proteolysis'. An enzyme dosage of 4%, a temperature of 50°, and a hydrolysis time of 300 min were found to be the optimum conditions to obtain high NR (84.28%) and degree of hydrolysis (DH, 16.18%) by orthogonal experiments. Glutamic acid was the most abundant amino acid of MDP (defatted mackerel protein) and MDPH (defatted mackerel protein hydrolysates). Compared with the FAO/WHO reference protein, the essential amino acid chemical scores (CS) were greater than 1.0 (1.0-1.7) in MDPH, which is reflective of high nutritional value. This, coupled with the light color and slight fishy odor, indicates that MDPH would potentially have a wide range of applications such as nutritional additives, functional ingredients, and so on.

  2. Artefact formation during acid hydrolysis of saponins from Medicago spp. (United States)

    Tava, Aldo; Biazzi, Elisa; Mella, Mariella; Quadrelli, Paolo; Avato, Pinarosa


    Artefact compounds obtained during acid hydrolysis of saponins from Medicago spp. (Fabaceae), have been monitored and evaluated by GC-FID. Their identification has been performed by GC-MS and 1H and 13C NMR. Saponins with different substituents on the triterpenic pentacyclic aglycones were considered, and their hydrolysis products were detected and quantified during 10 h of time course reaction. From soyasapogenol B glycoside the well known soyasapogenols B, C, D and F were obtained together with a previously undescribed sapogenol artefact identified as 3β,22β,24-trihydroxyolean-18(19)-en and named soyasapogenol H. From a zanhic acid saponin two major artefact compounds identified as 2β,3β,16α-trihydroxyolean-13(18)-en-23,28-dioic acid and 2β,3β,16α-trihydroxyolean-28,13β-olide-23-oic acid were obtained, together with some zanhic acid. Other compounds, detected in very small amount in the reaction mixture, were also tentatively identified based on their GC-MS and UV spectra. The other most characteristic saponins in Medicago spp., hederagenin, bayogenin and medicagenic acid glycosides, under acidic condition of hydrolysis, released instead the correspondent aglycones and generated a negligible amount of artefacts. Nature of artefacts and mechanism of their formation, involving a stable tertiary carbocation, is here proposed and discussed for the first time. Copyright © 2017 Elsevier Ltd. All rights reserved.

  3. Effect of sodium polyacrylate on the hydrolysis of octacalcium phosphate. (United States)

    Bigi, A; Boanini, E; Falini, G; Panzavolta, S; Roveri, N


    Octacalcium phosphate (OCP) hydrolysis into hydroxyapatite (HA) has been investigated in aqueous solutions at different concentrations of sodium polyacrylate (NaPA). In the absence of the polyelectrolyte, OCP undergoes a complete transformation into HA in 48 h. The hydrolysis is inhibited by the polymer, which is significantly adsorbed on the crystals, up to about 22 wt.%. A polymer concentration of 10(-2) mM is sufficient to cause a partial inhibition of OCP to HA transformation, which is completely hindered at higher concentrations. The small platelet-like crystals in the TEM images of partially converted OCP can display electron diffraction patterns characteristic either of OCP single crystals or of polycrystalline HA, whereas the much bigger plate-like crystals exhibit diffraction patterns characteristic of OCP single crystals. The polyelectrolyte adsorption on OCP crystals is accompanied by an increase of their mean length and by a significant reduction of the coherence length of the perfect crystalline domains along the c-axis direction. It is suggested that the carboxylate-rich polyelectrolyte is adsorbed on the hydrated layer of the OCP (100) face, thus inhibiting its in situ hydrolysis into HA.

  4. Sucrose hydrolysis by thermostable immobilized inulinases from aspergillus ficuum. (United States)

    Ettalibi, M; Baratti, J C.


    The possibility of using thermostable inulinases from Aspergillus ficuum in place of invertase for sucrose hydrolysis was explored. The commercial inulinases preparation was immobilized onto porous glass beads by covalent coupling using activation by a silane reagent and glutaraldehyde before adding the enzyme. The immobilization steps were optimized resulting in a support with 5,440 IU/g of support (sucrose hydrolysis) that is 77% of the activity of the free enzyme. Enzymatic properties of the immobilized inulinases were similar to those of the free enzymes with optimum pH near pH 5.0. However, temperature where the activity was maximal was shifted of 10 degrees C due to better thermal stability after immobilization with similar activation energies. The curve of the effect of sucrose concentration on activity was bi-phasic. The first part, for sucrose concentrations lower than 0.3 M, followed Michaelis-Menten kinetics with apparent K(M) and Vm only slightly affected by immobilization. Substrate inhibition was observed at values from 0.3 to 2 M sucrose. Complete sucrose hydrolysis was obtained for batch reactors with 0.3 and 1 M sucrose solutions. In continuous packed-bed reactor 100% (for 0.3 M sucrose), 90% (1 M sucrose) or 80% sucrose conversion were observed at space velocities of 0.06-0.25 h(-1). The operational half-life of the immobilized inulinases at 50 degrees C with 2 M sucrose was 350 days.

  5. In vitro enzymic hydrolysis of chlorogenic acids in coffee. (United States)

    da Encarnação, Joana Amarante; Farrell, Tracy L; Ryder, Alexandra; Kraut, Nicolai U; Williamson, Gary


    Coffee is rich in quinic acid esters of phenolic acids (chlorogenic acids) but also contains some free phenolic acids. A proportion of phenolic acids appear in the blood rapidly after coffee consumption due to absorption in the small intestine. We investigated in vitro whether this appearance could potentially be derived from free phenolic acids in instant coffee or from hydrolysis of chlorogenic acids by pancreatic or brush border enzymes. We quantified six free phenolic acids in instant coffees using HPLC-DAD-mass spectrometry. The highest was caffeic acid, but all were present at low levels compared to the chlorogenic acids. Roasting and decaffeination significantly reduced free phenolic acid content. We estimated, using pharmacokinetic modelling with previously published data, that the contribution of these compounds to small intestinal absorption is minimal. Hydrolysis of certain chlorogenic acids was observed with human-differentiated Caco-2 cell monolayers and with porcine pancreatin, which showed maximal rates on 3- and 5-O-caffeoylquinic acids, respectively. The amounts of certain free phenolic acids in coffee could only minimally account for small intestinal absorption based on modelling. The hydrolysis of caffeoylquinic, but not feruloylquinic acids, by enterocyte and pancreatic esterases is potentially a contributing mechanism to small intestinal absorption. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. Optimization of Enzymatic Hydrolysis of Waste Bread before Fermentation

    Directory of Open Access Journals (Sweden)

    Helena Hudečková


    Full Text Available Finding of optimal hydrolysis conditions is important for increasing the yield of saccharides. The higher yield of saccharides is usable for increase of the following fermentation effectivity. In this study optimal conditions (pH and temperature for amylolytic enzymes were searched. As raw material was used waste bread. Two analytical methods for analysis were used. Efficiency and process of hydrolysis was analysed spectrophotometrically by Somogyi-Nelson method. Final yields of glucose were analysed by HPLC. As raw material was used waste bread from local cafe. Waste bread was pretreated by grinding into small particles. Hydrolysis was performed in 100 mL of 15 % (w/v waste bread particles in the form of water suspension. Waste bread was hydrolysed by two commercial enzymes. For the liquefaction was used α‑amylase (BAN 240 L. The saccharification was performed by glucoamylase (AMG 300 L. Optimal conditions for α‑amylase (pH 6; 80 °C were found. The yield of total sugars was 67.08 g∙L-1 (calculated to maltose. As optimal conditions for glucoamylase (pH 4.2; 60 °C were found. Amount of glucose was 70.28 g∙L1. The time of waste bread liquefaction was 180 minutes. The time of saccharification was 90 minutes. The results were presented at the conference CECE Junior 2014.

  7. Lactose hydrolysis and milk powder production: technological aspects

    Directory of Open Access Journals (Sweden)

    Jansen Kelis Ferreira Torres


    Full Text Available The food industry has the challenge and the opportunity to develop new products with reduced or low lactose content in order to meet the needs of a growing mass of people with lactose intolerance. The manufacture of spray dried products with hydrolyzed lactose is extremely challenging. These products are highly hygroscopic, which influence the productivity and conservation of the powders, not to mention the undesirable and inevitable technological problem of constant clogging of drying chambers. The aim of this study was to evaluate the effect of different levels (0%, 25%, 50%, 75% and > 99% of enzymatic lactose hydrolysis on the production and storage of whole milk powder. The samples were processed in a pilot plant and characterized in relation to their composition analysis; to their degree of hydrolysis of lactose; and to their sorption isotherms. The results indicated the hydrolysis of lactose may affect the milk powder production due to a higher extent of powder adhesion within the spray dryer chambers and due to a higher tendency to absorb water during storage.

  8. Treatment of heterotopic ossification through remote ATP hydrolysis. (United States)

    Peterson, Jonathan R; De La Rosa, Sara; Eboda, Oluwatobi; Cilwa, Katherine E; Agarwal, Shailesh; Buchman, Steven R; Cederna, Paul S; Xi, Chuanwu; Morris, Michael D; Herndon, David N; Xiao, Wenzhong; Tompkins, Ronald G; Krebsbach, Paul H; Wang, Stewart C; Levi, Benjamin


    Heterotopic ossification (HO) is the pathologic development of ectopic bone in soft tissues because of a local or systemic inflammatory insult, such as burn injury or trauma. In HO, mesenchymal stem cells (MSCs) are inappropriately activated to undergo osteogenic differentiation. Through the correlation of in vitro assays and in vivo studies (dorsal scald burn with Achilles tenotomy), we have shown that burn injury enhances the osteogenic potential of MSCs and causes ectopic endochondral heterotopic bone formation and functional contractures through bone morphogenetic protein-mediated canonical SMAD signaling. We further demonstrated a prevention strategy for HO through adenosine triphosphate (ATP) hydrolysis at the burn site using apyrase. Burn site apyrase treatment decreased ATP, increased adenosine 3',5'-monophosphate, and decreased phosphorylation of SMAD1/5/8 in MSCs in vitro. This ATP hydrolysis also decreased HO formation and mitigated functional impairment in vivo. Similarly, selective inhibition of SMAD1/5/8 phosphorylation with LDN-193189 decreased HO formation and increased range of motion at the injury site in our burn model in vivo. Our results suggest that burn injury-exacerbated HO formation can be treated through therapeutics that target burn site ATP hydrolysis and modulation of SMAD1/5/8 phosphorylation. Copyright © 2014, American Association for the Advancement of Science.

  9. Complex enzyme hydrolysis releases antioxidative phenolics from rice bran. (United States)

    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. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. Hydrolysis of iron and chromium fluorides: mechanism and kinetics. (United States)

    Gálvez, José L; Dufour, Javier; Negro, Carlos; López-Mateos, Federico


    Fluoride complexes of metallic ions are one of the main problems when processing industrial effluents with high content of fluoride anion. The most important case is derived from pickling treatment of stainless steel, which is performed with HNO3/HF mixtures to remove oxides scale formed over the metal surface. Waste from this process, spent pickling liquor, must be treated for recovering metallic and acid content. Conventional treatments produce a final effluent with high quantity of fluoride complexes of iron and chromium. This work proposes a hydrolysis treatment of these solid metal fluorides by reacting them with a basic agent. Metal oxides are obtained, while fluoride is released to solution as a solved salt, which can be easily recovered as hydrofluoric acid. Solid iron and chromium fluorides, mainly K2FeF5(s) and CrF3(s), obtained in the UCM treatment process, were employed in this work. Optimal hydrolysis operating conditions were obtained by means of a factorial design: media must be basic but pH cannot be higher than 9.5, temperature from 40 to 70 degrees C and alkali concentration (potassium hydroxide) below 1.1 mol L(-1). Secondary reactions have been detected, which are probably due to fluoride adsorption onto obtained oxides surface. Mechanism of reaction consists of several stages, involving solid fluoride dissolution and complexes decomposition. Hydrolysis kinetics has been modeled with classical crystal dissolution kinetics, based on mass transfer phenomena.

  11. Stochastic molecular model of enzymatic hydrolysis of cellulose for ethanol production (United States)


    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 hydrolysis process. However, multiple factors affecting hydrolysis: cellulose structure and complex enzyme-substrate interactions during hydrolysis make it diffucult to develop mathematical kinetic models that can simulate hydrolysis in presence of multiple enzymes with high fidelity. In this study, a comprehensive hydrolysis model based on stochastic molecular modeling approch in which each hydrolysis event is translated into a discrete event is presented. The model captures the structural features of cellulose, enzyme properties (mode of actions, synergism, inhibition), and most importantly dynamic morphological changes in the substrate that directly affect the enzyme-substrate interactions during hydrolysis. Results Cellulose was modeled as a group of microfibrils consisting of elementary fibrils bundles, where each elementary fibril was represented as a three dimensional matrix of glucose molecules. Hydrolysis of cellulose was simulated based on Monte Carlo simulation technique. Cellulose hydrolysis results predicted by model simulations agree well with the experimental data from literature. Coefficients of determination for model predictions and experimental values were in the range of 0.75 to 0.96 for Avicel hydrolysis by CBH I action. Model was able to simulate the synergistic action of multiple enzymes during hydrolysis. The model simulations captured the important experimental observations: effect of structural properties, enzyme inhibition and enzyme loadings on the

  12. Stochastic molecular model of enzymatic hydrolysis of cellulose for ethanol production. (United States)

    Kumar, Deepak; Murthy, Ganti S


    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 hydrolysis process. However, multiple factors affecting hydrolysis: cellulose structure and complex enzyme-substrate interactions during hydrolysis make it diffucult to develop mathematical kinetic models that can simulate hydrolysis in presence of multiple enzymes with high fidelity. In this study, a comprehensive hydrolysis model based on stochastic molecular modeling approch in which each hydrolysis event is translated into a discrete event is presented. The model captures the structural features of cellulose, enzyme properties (mode of actions, synergism, inhibition), and most importantly dynamic morphological changes in the substrate that directly affect the enzyme-substrate interactions during hydrolysis. Cellulose was modeled as a group of microfibrils consisting of elementary fibrils bundles, where each elementary fibril was represented as a three dimensional matrix of glucose molecules. Hydrolysis of cellulose was simulated based on Monte Carlo simulation technique. Cellulose hydrolysis results predicted by model simulations agree well with the experimental data from literature. Coefficients of determination for model predictions and experimental values were in the range of 0.75 to 0.96 for Avicel hydrolysis by CBH I action. Model was able to simulate the synergistic action of multiple enzymes during hydrolysis. The model simulations captured the important experimental observations: effect of structural properties, enzyme inhibition and enzyme loadings on the hydrolysis and degree of

  13. Effects of Toasting Time on Digestive Hydrolysis of Soluble and Insoluble 00-Rapeseed Meal Proteins. (United States)

    Salazar-Villanea, Sergio; Bruininx, Erik M A M; Gruppen, Harry; Carré, Patrick; Quinsac, Alain; van der Poel, Antonius F B


    Thermal damage to proteins can reduce their nutritional value. The effects of toasting time on the kinetics of hydrolysis, the resulting molecular weight distribution of 00-rapeseed meal (RSM) and the soluble and insoluble protein fractions separated from the RSM were studied. Hydrolysis was performed with pancreatic proteases to represent in vitro protein digestibility. Increasing the toasting time of RSM linearly decreased the rate of protein hydrolysis of RSM and the insoluble protein fractions. The extent of hydrolysis was, on average, 44% higher for the insoluble compared with the soluble protein fraction. In contrast, the rate of protein hydrolysis of the soluble protein fraction was 3-9-fold higher than that of the insoluble protein fraction. The rate of hydrolysis of the insoluble protein fraction linearly decreased by more than 60% when comparing the untoasted to the 120 min toasted RSM. Increasing the toasting time elicited the formation of Maillard reaction products (furosine, N(ε)-carboxymethyl-lysine and N(ε)-carboxyethyl-lysine) and disulfide bonds in the insoluble protein fraction, which is proposed to explain the reduction in the hydrolysis rate of this fraction. Overall, longer toasting times increased the size of the peptides resulting after hydrolysis of the RSM and the insoluble protein fraction. The hydrolysis kinetics of the soluble and insoluble protein fractions and the proportion of soluble:insoluble proteins in the RSM explain the reduction in the rate of protein hydrolysis observed in the RSM with increasing toasting time.

  14. Improving the hydrolysis efficiency of soy sauce residue using ultrasonic probe-assisted enzymolysis technology. (United States)

    Chen, Xiangrong; Luo, Yijie; Qi, Benkun; Luo, Jianquan; Wan, Yinhua


    Ultrasonic probe-assisted enzymolysis technology was developed to improve the hydrolysis efficiency of soy sauce residue (SSR). The effects of enzyme type and enzymatic hydrolysis parameters on the hydrolysis degree of SSR were studied firstly to obtain the optimal enzymatic hydrolysis conditions. Then the effects of ultrasound on protease activity and structure of SSR were investigated to elucidate the acting mechanism of ultrasound. Finally, the ultrasonic-assisted enzymatic hydrolysis modes were designed and compared, and the hydrolysates from SSR were characterized to evaluate their further application. The results showed that a hydrolysis degree of 15.53% could be obtained under the optimum enzymolysis conditions: enzyme amount 6000U/g, pH 7.8, temperature 50°C, the ratio of substrate to water phase 1:20, hydrolysis time 4h. Increasing ultrasound treatment time or power could reduce substrate size and consequently enhance the catalytic surface area. Prolonging ultrasound treatment time had a negative influence on enzyme activity, but low ultrasound power was helpful for increasing the enzyme activity. Ultrasound pretreatment of SSR followed by enzymatic hydrolysis increased the hydrolysis degree by 47.6%. When the ultrasound was applied directly to enzymolysis process, the hydrolysis degree of SSR exhibited an increase of 33.0%. The hydrolysates from SSR exhibited good antioxidant activities, and had a potential use as a functional ingredient in food or feed industry. Copyright © 2016 Elsevier B.V. All rights reserved.

  15. Reaction pathways and free energy profiles for spontaneous hydrolysis of urea and tetramethylurea: Unexpected substituent effects (United States)

    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 competing non-enzymatic hydrolysis pathways have demonstrated that the dominant pathway is the neutral hydrolysis via the CN addition for both urea (when pHhydrolysis of amides where alkaline hydrolysis is dominant. Based on the computational data, the substituent shift of free energy barrier calculated for the neutral hydrolysis is remarkably different from that for the alkaline hydrolysis, and the rate constant for the urea hydrolysis should be ~1.3×109-fold lower than that (4.2×10−12 s−1) measured for the Me4U hydrolysis. As a result, the rate enhancement and catalytic proficiency of urease should be 1.2×1025 and 3×1027 M−1, respectively, suggesting that urease surpasses proteases and all other enzymes in its power to enhance the rate of reaction. All of the computational results are consistent with available experimental data for Me4U, suggesting that the computational prediction for urea is reliable. PMID:24097048

  16. Hydrolysis of nitriles by soil bacteria: variation with soil origin. (United States)

    Rapheeha, O K L; Roux-van der Merwe, M P; Badenhorst, J; Chhiba, V; Bode, M L; Mathiba, K; Brady, D


    The aim of this study was to explore bacterial soil diversity for nitrile biocatalysts, in particular, those for hydrolysis of β-substituted nitriles, to the corresponding carboxamides and acids that may be incorporated into peptidomimetics. To achieve this, we needed to compare the efficiency of isolation methods and determine the influence of land use and geographical origin of the soil sample. Nitrile-utilizing bacteria were isolated from various soil environments across a 1000 km long transect of South Africa, including agricultural soil, a gold mine tailing dam and uncultivated soil. The substrate profile of these isolates was determined through element-limited growth studies on seven different aliphatic or aromatic nitriles. A subset of these organisms expressing broad substrate ranges was evaluated for their ability to hydrolyse β-substituted nitriles (3-amino-3-phenylpropionitrile and 3-hydroxy-4-phenoxybutyronitrile) and the active organisms were found to be Rhodococcus erythropolis from uncultivated soil and Rhodococcus rhodochrous from agricultural soils. The capacity for hydrolysis of β-substituted nitriles appears to reside almost exclusively in Rhodococci. Land use has a much greater effect on the biocatalysis substrate profile than geographical location. Enzymes are typically substrate specific in their catalytic reactions, and this means that a wide diversity of enzymes is required to provide a comprehensive biocatalysis toolbox. This paper shows that the microbial diversity of nitrile hydrolysis activity can be targeted according to land utilization. Nitrile biocatalysis is a green chemical method for the enzymatic production of amides and carboxylic acids that has industrial applications, such as in the synthesis of acrylamide and nicotinamide. The biocatalysts discovered in this study may be applied to the synthesis of peptidomimetics which are an important class of therapeutic compounds. © 2016 The Society for Applied Microbiology.

  17. Maltodextrin hydrolysis in a fluidized-bed immobilized enzyme reactor

    Energy Technology Data Exchange (ETDEWEB)

    Vallat, I.; Monsan, P.; Riba, J.P.


    The present work deals with maltodextrin hydrolysis by glucoamylase immobilized onto corn stover in a fluidized bed reactor. An industrial enzyme preparation was convalently grafted onto corn stover, yielding an activity of up to 372 U/g and 1700 U/g for support particle sizes of 0.8 and 0.2 mm, respectively. A detailed kinetic study, using a differntial reactor, allowed the characterization of the influence of mass transfer resistance on the reaction catalyzed by immobilized glucoamylase. A simple and general mathematical model was then developed to describe the experimental conversion data and found to be vaild.

  18. A comparative study of the hydrolysis of gamma irradiated lignocelluloses

    Directory of Open Access Journals (Sweden)

    E. Betiku


    Full Text Available The effect of high-dose irradiation as a pretreatment method on two common lignocellulosic materials; hardwood (Khaya senegalensis and softwood (Triplochiton scleroxylon were investigated by assessing the potential of cellulase enzyme derived from Aspergillus flavus Linn isolate NSPR 101 to hydrolyse the materials. The irradiation strongly affected the materials, causing the enzymatic hydrolysis to increase by more than 3 fold. Maximum digestibility occurred in softwood at 40kGy dosage of irradiation, while in hardwood it was at 90kGy dosage. The results also showed that, at the same dosage levels (p < 0.05, hardwood was hydrolysed significantly better compared to the softwood.

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

    Directory of Open Access Journals (Sweden)

    Galbe Mats


    Full Text Available 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 leads to more compact solids, which may affect subsequent processing steps. This study was carried out to investigate the influence of pressing the biomass, in combination with the effects of not washing the material, on the sugar yield obtained from two-step dilute acid hydrolysis, with and without subsequent enzymatic digestion of the solids. Results Washing the material between the two acid hydrolysis steps, followed by enzymatic digestion, resulted in recovery of 96% of the mannose and 81% of the glucose (% of the theoretical in the liquid fraction, regardless of the choice of dewatering method (pressing or vacuum filtration. Not washing the solids between the two acid hydrolysis steps led to elevated acidity of the remaining solids during the second hydrolysis step, which resulted in lower yields of mannose, 85% and 74% of the theoretical, for the pressed and vacuum-filtered slurry, respectively, due to sugar degradation. However, this increase in acidity resulted in a higher glucose yield (94.2% from pressed slurry than from filtered slurry (77.6%. Conclusion Pressing the washed material between the two acid hydrolysis steps had no significant negative effect on the sugar yields of the second acid hydrolysis step or on enzymatic hydrolysis. Not washing the material resulted in a harsher second acid hydrolysis step, which caused greater degradation of the sugars during subsequent acid hydrolysis of the solids, particularly in case of the vacuum

  20. Hydrolysis of tanned leather wastes under alkaline, acidic and oxidative conditions

    Directory of Open Access Journals (Sweden)

    Botić Tatjana


    Full Text Available Different wastes in large quantities are the outcome or the by-product of processes in the tanning industry. The largest part of solid wastes is collagen based and obtained in the reprocessing steps of tanned and non-tanned products. The quality collagen substance obtained from wastes of the leather industry used as a supporting material in many reprocessing industries. Hydrolysis is the basic step of collagen processing, namely the shortening of its polypeptide chain. The main goal of this investigation was to examine the influence of the following parameters on the extent of collagen hydrolysis: type and quantity of reagent used for hydrolysis temperature of hydrolysis, pH, duration of hydrolysis as well as the dimensions of the "shavings" samples. The change of molar mass during hydrolysis was monitored by UV-spectroscopy. Special attention was paid to the dechroming process of the collagen hydrolysate.

  1. Fuzzy logic feedback control for fed-batch enzymatic hydrolysis of lignocellulosic biomass. (United States)

    Tai, Chao; Voltan, Diego S; Keshwani, Deepak R; Meyer, George E; Kuhar, Pankaj S


    A fuzzy logic feedback control system was developed for process monitoring and feeding control in fed-batch enzymatic hydrolysis of a lignocellulosic biomass, dilute acid-pretreated corn stover. Digested glucose from hydrolysis reaction was assigned as input while doser feeding time and speed of pretreated biomass were responses from fuzzy logic control system. Membership functions for these three variables and rule-base were created based on batch hydrolysis data. The system response was first tested in LabVIEW environment then the performance was evaluated through real-time hydrolysis reaction. The feeding operations were determined timely by fuzzy logic control system and efficient responses were shown to plateau phases during hydrolysis. Feeding of proper amount of cellulose and maintaining solids content was well balanced. Fuzzy logic proved to be a robust and effective online feeding control tool for fed-batch enzymatic hydrolysis.

  2. Enhancing saccharification of cassava stems by starch hydrolysis prior to pretreatment


    Martín, Carlos; Wei, Maogui; Xiong, Shaojun; Jönsson, Leif J


    Chemical characterization of cassava stems from different origin revealed that glucans accounted for 54-63% of the dry weight, whereas 35-67% of these glucans consisted of starch. The cassava stems were subjected to a saccharification study including starch hydrolysis, pretreatment with either sulfuric acid or 1-ethyl-3-methylimidazolium acetate ([Emim]OAc), and enzymatic hydrolysis of cellulose. Starch hydrolysis prior to pretreatment decreased sugar degradation, improved enzymatic convertib...

  3. Effects of Toasting Time on Digestive Hydrolysis of Soluble and Insoluble 00-Rapeseed Meal Proteins


    Salazar-Villanea, Sergio; Bruininx, Erik M.A.M.; Gruppen, Harry; Carré, Patrick; Quinsac, Alain; Poel, de, K.R.


    Thermal damage to proteins can reduce their nutritional value. The effects of toasting time on the kinetics of hydrolysis, the resulting molecular weight distribution of 00-rapeseed meal (RSM) and the soluble and insoluble protein fractions separated from the RSM were studied. Hydrolysis was performed with pancreatic proteases to represent in vitro protein digestibility. Increasing the toasting time of RSM linearly decreased the rate of protein hydrolysis of RSM and the insoluble protein frac...

  4. Hydrolysis of particulate substrate by activated sludge under aerobic, anoxic and anaerobic conditions

    DEFF Research Database (Denmark)

    Henze, Mogens; Mladenovski, C.


    , anoxic and anaerobic conditions, the hydrolysis was following through the production of ammonia. The hydrolysis rate of nitrogeneous compounds is significantly affected by the electron donor available. The rate is high under aerobic conditions, medium under anaerobic conditions and low under anoxic...... conditions. The ratio between the hydrolysis rates under aerobic and under anoxic conditions are very similar to the respiration rates measured as electron equivalents....

  5. [Study on optimization of the hydrolysis process of dioscin catalyzed by hydrochloric acid]. (United States)

    Li, Qian; Gao, Xiang-Tao; Zhao, Wen-Lei; Liu, Xiu-Hua


    To study the optimization of hydrolysis process of dioscin catalyzed by hydrochloric acid. The effects of hydrochloric acid concentration, reaction temperature, reaction time and their interactions had been investigated using orthogonal design and single-factor experiments. The optimum hydrolysis conditions were as follows: reaction temperature 90 degrees C, reaction time 3h, hydrochloric acid concentration 2.0 mol/L. Hydrochloric acid concentration and reaction temperature should be controlled because of their remarkable influence on hydrolysis process.

  6. Validation of Inhibition Effect in the Cellulose Hydrolysis: a Dynamic Modelling Approach

    DEFF Research Database (Denmark)

    Morales Rodriguez, Ricardo; Tsai, Chien-Tai; Meyer, Anne S.


    Enzymatic hydrolysis is one of the main steps in the processing of bioethanol from lignocellulosic raw materials. However, complete understanding of the underlying phenomena is still under development. Hence, this study has focused on validation of the inhibition effects in the cellulosic biomass...... hydrolysis employing a dynamic mathematical model. A systematic framework for parameter estimation is used for model validation, which helps overcome the problem of parameter correlation. Data sets obtained from carefully designed enzymatic cellulose and cellobiose hydrolysis experiments, were used...

  7. Glucose obtained from rice bran by ultrasound-assisted enzymatic hydrolysis


    Raquel Cristine Kuhn; Marcio Antonio Mazutti; Edson Luiz Foletto; Valéria Dal Prá; Eduardo Zimmermann; Matheus Souza; Vitória Segabinazzi Foletto; Tanisa Paula Silveira Maleski; Felipe Cavalheiro Lunelli; Pâmela Sfalcin


    In this work ultrasound-assisted solid-state enzymatic hydrolysis of rice bran to obtain fermentable sugars was investigated. For this purpose, process variables such as temperature, enzyme concentration and moisture content were evaluated during the enzymatic hydrolysis with and without ultrasound irradiation. The enzyme used is a blend of amylases derived from genetically modified strains of Trichoderma reesei. Kinetic of the enzymatic hydrolysis of rice bran at the constant-reaction rate p...

  8. Trace metals in humic acids and their hydrolysis products

    Energy Technology Data Exchange (ETDEWEB)

    Huljev, D.J.


    The role of humic acids in the transport of trace elements in the Sava river biocycle (near Zagreb, Yugoslavia) has been investigated by means of neutron activation analysis. Humic acids can accumulate certain trace elements, but little is known about their selectivity under certain physicochemical conditions. Humic acids isolated from Sava river sediments were hydrolized to their main components, namely to amino acids (proteins), phenols (and phenolic acids), carbohydrates, polycyclic aromatics, and metals. Degradation of humic acids in the environment occurs by the action of microorganisms, and during this step, certain trace metals become available to Sava river animals through standard physiological processes. In this work neutron activation analysis was applied to the quantitative and qualitative determination of trace metals in humic acids of Sava river origin, and in their hydrolysis products. The trace metals measured were Ag, Fe, Co, Cs, Eu, Sb, Sc, Se, and Sr. The concentrations obtained ranged from 0.05 to 2730 of a certain microconstituent per gram of sample. The experimental data are compared with laboratory experiments on the determination of stability constants for metal complexes of humic acids and their hydrolysis product. From these data it is possible to suggest the ecological importance of trace metals bonded to humic acids in a certain region. The results concerning the role of humic acids in the cycle of trace elements in Sava river biocycle are discussed.

  9. Carbon-based strong solid acid for cornstarch hydrolysis

    Energy Technology Data Exchange (ETDEWEB)

    Nata, Iryanti Fatyasari, E-mail: [Chemical Engineering Study Program, Faculty of Engineering, Lambung Mangkurat University, Jl. A. Yani Km. 36 Banjarbaru, South Kalimantan 70714 (Indonesia); Irawan, Chairul; Mardina, Primata [Chemical Engineering Study Program, Faculty of Engineering, Lambung Mangkurat University, Jl. A. Yani Km. 36 Banjarbaru, South Kalimantan 70714 (Indonesia); Lee, Cheng-Kang, E-mail: [Department of Chemical Engineering, National Taiwan University of Science and Technology, 43 Keelung Rd. Sec.4, Taipei 106, Taiwan (China)


    Highly sulfonated carbonaceous spheres with diameter of 100–500 nm can be generated by hydrothermal carbonization of glucose in the presence of hydroxyethylsulfonic acid and acrylic acid at 180 °C for 4 h. The acidity of the prepared carbonaceous sphere C4-SO{sub 3}H can reach 2.10 mmol/g. It was used as a solid acid catalyst for the hydrolysis of cornstarch. Total reducing sugar (TRS) concentration of 19.91 mg/mL could be obtained by hydrolyzing 20 mg/mL cornstarch at 150 °C for 6 h using C4-SO{sub 3}H as solid acid catalyst. The solid acid catalyst demonstrated good stability that only 9% decrease in TRS concentration was observed after five repeat uses. The as-prepared carbon-based solid acid catalyst can be an environmentally benign replacement for homogeneous catalyst. - Highlights: • Carbon solid acid was successfully prepared by one-step hydrothermal carbonization. • The acrylic acid as monomer was effectively reduce the diameter size of particle. • The solid acid catalyst show good catalytic performance of starch hydrolysis. • The solid acid catalyst is not significantly deteriorated after repeated use.

  10. High-risk biodegradable waste processing by alkaline hydrolysis. (United States)

    Kalambura, Sanja; Voća, Neven; Krička, Tajana; Sindrak, Zoran; Spehar, Ana; Kalambura, Dejan


    Biodegradable waste is by definition degraded by other living organisms. Every day, meat industry produces large amounts of a specific type of biodegradable waste called slaughterhouse waste. Traditionally in Europe, this waste is recycled in rendering plants which produce meat and bone meal and fat. However, feeding animals with meat and bone meal has been banned since the outbreaks of bovine spongiform encephalopathy (BSE). In consequence, new slaughterhouse waste processing technologies have been developed, and animal wastes have now been used for energy production. Certain parts of this waste, such as brains and spinal cord, are deemed high-risk substances, because they may be infected with prions. Their treatment is therefore possible only in strictly controlled conditions. One of the methods which seems to bear acceptable health risk is alkaline hydrolysis. This paper presents the results of an alkaline hydrolysis efficiency study. It also proposes reuse of the obtained material as organic fertiliser, as is suggested by the analytical comparison between meat and bone meal and hydrolysate.

  11. New bioreactor for fibrous support and hydrolysis of penicillin G

    Energy Technology Data Exchange (ETDEWEB)

    Ishimura, F. [Asahi Chemical Industry Co. Ltd., Tokyo (Japan)


    Phenylacetic acid, a strong acid was formed from the enzymatic hydrolysis of penicillin G to 6-amino-peni-cillanic acid. Because of the difficulty of maintaining the pH constant inside the reactor the reaction mixture was circulated at a high flow rate between the packed bed reactor and the vessel for adjusting phi The reactor and the vessel see connected via a pump. When an immobilized enzyme made of compressible fibers was packed in the packed bed reactor the hydrolytic efficiency of the immobilized enzyme was approximately 0.5. The low efficiency of the immobilized enzyme was assumed to be caused by the aggregation of the immobilized enzyme due to the pressure generated by flow of the reaction mixture. Therefore the aggregation in the packed bed reactor was analyzed experimentally using Kozeny- Carman`s equation in order to estimate the distribution of the aggregation in the reactor. Furthermore a new column devised specially to prevent aggregation of the compressible fibrous support was developed. A reaction time required for completing the hydrolysis (98 % of conversion) was adopted to estimate the efficiency of the entire immobilized enzyme packed in the reactor throughout the entire period of reaction. The efficiency of the immobilized enzyme packed in the new column of the reactor was concluded to be increased by approximately 1.8 times, compared with that of conventional packed bed columns. 5 refs., 9 figs., 1 tab.

  12. Hydrolysis kinetics of tulip tree xylan in hot compressed water. (United States)

    Yoon, Junho; Lee, Hun Wook; Sim, Seungjae; Myint, Aye Aye; Park, Hee Jeong; Lee, Youn-Woo


    Lignocellulosic biomass, a promising renewable resource, can be converted into numerous valuable chemicals post enzymatic saccharification. However, the efficacy of enzymatic saccharification of lignocellulosic biomass is low; therefore, pretreatment is necessary to improve the efficiency. Here, a kinetic analysis was carried out on xylan hydrolysis, after hot compressed water pretreatment of the lignocellulosic biomass conducted at 180-220°C for 5-30min, and on subsequent xylooligosaccharide hydrolysis. The weight ratio of fast-reacting xylan to slow-reacting xylan was 5.25 in tulip tree. Our kinetic results were applied to three different reaction systems to improve the pretreatment efficiency. We found that semi-continuous reactor is promising. Lower reaction temperatures and shorter space times in semi-continuous reactor are recommended for improving xylan conversion and xylooligosaccharide yield. In the theoretical calculation, 95% of xylooligosaccharide yield and xylan conversion were achieved simultaneously with high selectivity (desired product/undesired product) of 100 or more. Copyright © 2016. Published by Elsevier Ltd.

  13. Hydrolysis by Alcalase Improves Hypoallergenic Properties of Goat Milk Protein. (United States)

    Jung, Tae-Hwan; Yun, Sung-Seob; Lee, Won-Jae; Kim, Jin-Wook; Ha, Ho-Kyung; Yoo, Michelle; Hwang, Hyo-Jeong; Jeon, Woo-Min; Han, Kyoung-Sik


    Goat milk is highly nutritious and is consumed in many countries, but the development of functional foods from goat milk has been slow compared to that for other types of milk. The aim of this study was to develop a goat milk protein hydrolysate (GMPH) with enhanced digestibility and better hypoallergenic properties in comparison with other protein sources such as ovalbumin and soy protein. Goat milk protein was digested with four commercial food-grade proteases (separately) under various conditions to achieve the best hydrolysis of αs -casein and β-lactoglobulin. It was shown that treatment with alcalase (0.4%, 60℃ for 30 min) effectively degraded these two proteins, as determined by SDS-PAGE, measurement of nonprotein nitrogen content, and reverse-phase high-performance liquid chromatography. Hydrolysis with alcalase resulted in a significant decrease in β-lactoglobulin concentration (almost to nil) and a ~40% reduction in the level of αs-casein. Quantification of histamine and TNF-α released from HMC-1 cells (human mast cell line) showed that the GMPH did not induce an allergic response when compared to the control. Hence, the GMPH may be useful for development of novel foods for infants, the elderly, and convalescent patients, to replace cow milk.

  14. Hydrolysis of dinitrobenzamide phosphate prodrugs: the role of alkaline phosphatase. (United States)

    Lo, Wing-Yee; Balasubramanian, Amit; Helsby, Nuala A


    Phosphate prodrugs which undergo hydrolysis in vivo have been used to improve the solubility and pharmacokinetic properties of a number of drugs. Dinitrobenzamide mustards (DNBM) are examples of such drugs. We investigated the ability of purified alkaline phosphatase isoforms to dephosphorylate three DNBM phosphate prodrugs. In addition, the relative rate of dephosphorylation of these phosphate prodrugs in a number of tissues was determined. These phosphate prodrugs are indeed substrates for alkaline phosphatase, with time dependent formation of the hydrolysis product. Intestinal alkaline phosphatase (IAP) and placental alkaline phosphatase (PLAP) had the highest activity for these substrates and compound P2 was the most rapidly metabolised. Similarly, compound P2 had the shortest half life in mouse serum (t1/2 = 1.15 h) compared with P1 (t1/2 = 13.34 h) and P3 (t1/2 = 4.4 h). However, serum has very low dephosphorylase activity for these substrates compared with intestine and liver homogenates. In addition, there is little or no difference in the relative rate of dephosphorylation of each of the three compounds in mouse tissues in contrast to the pattern observed with purified alkaline phosphatase and mouse serum. Hence additional phosphatase enzymes may be involved in the metabolism of phosphate prodrugs in vivo.

  15. Parameter and Process Significance in Mechanistic Modeling of Cellulose Hydrolysis (United States)

    Rotter, B.; Barry, A.; Gerhard, J.; Small, J.; Tahar, B.


    The rate of cellulose hydrolysis, and of associated microbial processes, is important in determining the stability of landfills and their potential impact on the environment, as well as associated time scales. To permit further exploration in this field, a process-based model of cellulose hydrolysis was developed. The model, which is relevant to both landfill and anaerobic digesters, includes a novel approach to biomass transfer between a cellulose-bound biofilm and biomass in the surrounding liquid. Model results highlight the significance of the bacterial colonization of cellulose particles by attachment through contact in solution. Simulations revealed that enhanced colonization, and therefore cellulose degradation, was associated with reduced cellulose particle size, higher biomass populations in solution, and increased cellulose-binding ability of the biomass. A sensitivity analysis of the system parameters revealed different sensitivities to model parameters for a typical landfill scenario versus that for an anaerobic digester. The results indicate that relative surface area of cellulose and proximity of hydrolyzing bacteria are key factors determining the cellulose degradation rate.

  16. Enhancement of hydrolysis of Chlorella vulgaris by hydrochloric acid. (United States)

    Park, Charnho; Lee, Ja Hyun; Yang, Xiaoguang; Yoo, Hah Young; Lee, Ju Hun; Lee, Soo Kweon; Kim, Seung Wook


    Chlorella vulgaris is considered as one of the potential sources of biomass for bio-based products because it consists of large amounts of carbohydrates. In this study, hydrothermal acid hydrolysis with five different acids (hydrochloric acid, nitric acid, peracetic acid, phosphoric acid, and sulfuric acid) was carried out to produce fermentable sugars (glucose, galactose). The hydrothermal acid hydrolysis by hydrochloric acid showed the highest sugar production. C. vulgaris was hydrolyzed with various concentrations of hydrochloric acid [0.5-10 % (w/w)] and microalgal biomass [20-140 g/L (w/v)] at 121 °C for 20 min. Among the concentrations examined, 2 % hydrochloric acid with 100 g/L biomass yielded the highest conversion of carbohydrates (92.5 %) into reducing sugars. The hydrolysate thus produced from C. vulgaris was fermented using the yeast Brettanomyces custersii H1-603 and obtained bioethanol yield of 0.37 g/g of algal sugars.

  17. Kinetic study of sulphuric acid hydrolysis of protein feathers. (United States)

    Ben Hamad Bouhamed, Sana; Kechaou, Nabil


    Poultry feather keratin is the most important by-product from the poultry industry due to its abundance. Different methods have been still applied to process this by-product such as enzymatic hydrolysis which is expensive and inapplicable at the industrial level. This paper presents a study of acid hydrolysis of poultry feathers using different types of acids, sulphuric acid concentration, different temperatures and solid to liquid ratio to obtain a liquid product rich in peptides. The feathers analysis revealed a crude protein content of 88.83%. A maximum peptides production of 676 mg/g was reached using sulphuric acid, 1 molar acid concentration and 50 g/l solid to liquid ratio at a temperature of 90 °C after 300 min. A reaction scheme for protein aggregation and decomposition to polypeptides and amino acids was proposed and a kinetic model for peptides production was developed. The proposed kinetic model proved to be well adapted to the experimental data with R (2) = 0.99.

  18. Base hydrolysis kinetics of HMX-based explosives using sodium carbonate

    Energy Technology Data Exchange (ETDEWEB)

    Bishop, R.L.; Skidmore, C.; Flesner, R.L.; Dell`orco, P.C.; Spontarelli, T.; Uher, K.J.; Kramer, J.F. [Los Alamos National Lab., NM (United States); Bell, D.A. [Wyoming Univ., Laramie, WY (United States)


    Sodium carbonate has been identified as a possible hydrolysis reagent for decomposing HMX-based explosives to water soluble, non-energetic products. In this study, the reaction kinetics of sodium carbonate hydrolysis are examined and a reaction model is developed. The rate of hydrolysis is reaction rate limited, opposed to mass transfer limited, up to 150{degrees}C. Greater than 99% of the explosive solids in powder form are destroyed in less than 10 minutes at a temperature of 150{degrees}C. The primary products from sodium carbonate hydrolysis are sodium nitrite, formate, nitrate, acetate, glycolate, hexamine, nitrogen gas, nitrous oxide, and ammonia.

  19. Linking Hydrolysis Performance to Trichoderma reesei Cellulolytic Enzyme Profile

    DEFF Research Database (Denmark)

    Lehmann, Linda Olkjær; Petersen, Nanna; I. Jørgensen, Christian


    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, e.g. by spiking with single enzymes and monitoring hydrolysis performance. In this study a multivariate...... approach, partial least squares regression, was used to see if it could help explain the correlation between enzyme profile and hydrolysis performance. Diverse enzyme mixtures were produced by Trichoderma reesei Rut-C30 by exploiting various fermentation conditions and used for hydrolysis of washed...

  20. Optimization of hydrolysis conditions for bovine plasma protein using response surface methodology. (United States)

    Seo, Hyun-Woo; Jung, Eun-Young; Go, Gwang-Woong; Kim, Gap-Don; Joo, Seon-Tea; Yang, Han-Sul


    The purpose of this study was to establish optimal conditions for the hydrolysis of bovine plasma protein. Response surface methodology was used to model and optimize responses [degree of hydrolysis (DH), 2,2-diphenyl-1-picrydrazyl (DPPH) radical-scavenging activity and Fe(2+)-chelating activity]. Hydrolysis conditions, such as hydrolysis temperature (46.6-63.4 °C), hydrolysis time (98-502 min), and hydrolysis pH (6.32-9.68) were selected as the main processing conditions in the hydrolysis of bovine plasma protein. Optimal conditions for maximum DH (%), DPPH radical-scavenging activity (%) and Fe(2+)-chelating activity (%) of the hydrolyzed bovine plasma protein, were respectively established. We discovered the following three conditions for optimal hydrolysis of bovine plasma: pH of 7.82-8.32, temperature of 54.1 °C, and time of 338.4-398.4 min. We consequently succeeded in hydrolyzing bovine plasma protein under these conditions and confirmed the various desirable properties of optimal hydrolysis. Copyright © 2015 Elsevier Ltd. All rights reserved.

  1. Symmetry broken and rebroken during the ATP hydrolysis cycle of the mitochondrial Hsp90 TRAP1. (United States)

    Elnatan, Daniel; Betegon, Miguel; Liu, Yanxin; Ramelot, Theresa; Kennedy, Michael A; Agard, David A


    Hsp90 is a homodimeric ATP-dependent molecular chaperone that remodels its substrate 'client' proteins, facilitating their folding and activating them for biological function. Despite decades of research, the mechanism connecting ATP hydrolysis and chaperone function remains elusive. Particularly puzzling has been the apparent lack of cooperativity in hydrolysis of the ATP in each protomer. A crystal structure of the mitochondrial Hsp90, TRAP1, revealed that the catalytically active state is closed in a highly strained asymmetric conformation. This asymmetry, unobserved in other Hsp90 homologs, is due to buckling of one of the protomers and is most pronounced at the broadly conserved client-binding region. Here, we show that rather than being cooperative or independent, ATP hydrolysis on the two protomers is sequential and deterministic. Moreover, dimer asymmetry sets up differential hydrolysis rates for each protomer, such that the buckled conformation favors ATP hydrolysis. Remarkably, after the first hydrolysis, the dimer undergoes a flip in the asymmetry while remaining in a closed state for the second hydrolysis. From these results, we propose a model where direct coupling of ATP hydrolysis and conformational flipping rearranges client-binding sites, providing a paradigm of how energy from ATP hydrolysis can be used for client remodeling.

  2. Impact of α-amylase combined with hydrochloric acid hydrolysis on structure and digestion of waxy rice starch. (United States)

    Li, Hongyan; Zhu, Yanqiao; Jiao, Aiquan; Zhao, Jianwei; Chen, Xiaoming; Wei, Benxi; Hu, Xiuting; Wu, Chunsen; Jin, Zhengyu; Tian, Yaoqi


    The structure and in vitro digestibility of native waxy rice starch by the combined hydrolysis of α-amylase and hydrochloric acid were investigated in this study. The combined hydrolysis technique generated higher hydrolysis rate and extent than the enzymatic hydrolysis. The granular appearance and chromatograph profile demonstrated that α-amylase and hydrochloric acid exhibited different patterns of hydrolysis. The rise in the ratio of absorbance 1047/1022cm(-1), the melting temperature range (Tc-To), and the melting enthalpy (ΔH) were observed during the combined hydrolysis. These results suggest that α-amylase simultaneously cleaves the amorphous and crystalline regions, whereas the amorphous regions of starch granules are preferentially hydrolyzed during the acid hydrolysis. Furthermore, the combined hydrolysis increased rapidly digestible starch (RDS) while decreased slowly digestible starch (SDS) and resistant starch (RS), indicating that the hydrolysis mode affected the digestion property of native waxy rice starch. Copyright © 2013 Elsevier B.V. All rights reserved.

  3. Enzymic hydrolysis of wood and straw polysaccharides. 10. Effect of the morphological structure of cell walls on hydrolysis. [Cellulase, Cellokoningin P10 X, Celloviridin G3X, Cellolignorin

    Energy Technology Data Exchange (ETDEWEB)

    Katkevich, Y.Y.; Gromov, V.S.; Vevere, P.; Samokhvalova, T.A.


    Treatment of powdered wood and straw with cellulolytic enzymes (cellulase, Cellokoningin P10 X, Celloviridin G3X, and Cellolignorin) resulted in the degradation of cell wall biopolymers and facilitated the hydrolysis of polysaccharides of the powdered material. The rate of hydrolysis depended on the high degree of disintegration of wood (from aspen and spruce), straw (wheat and rye), and cellulose sulfate (from henbane). The outer layers of cellulose fibers were most resistant to cellulases. Kinetics of the enzymic reactions are given.

  4. Kinetic and thermal characterization of the hydrolysis of polysuccinimide

    Energy Technology Data Exchange (ETDEWEB)

    Mosig, J.; Gooding, C.H.; Wheeler, A.P. [Clemson Univ., SC (United States)


    Hundreds of millions of pounds of anionic polymers, particularly poly(acrylate) and its derivatives, are used annually as dispersants and antiscalants in water-treatment formulations and detergents, and much of this is released into environmental waters. Though these polymers are apparently not toxic, neither are they especially biodegradable. The kinetics of the base hydrolysis of polysuccinimide in an aqueous slurry were studied with temperatures ranging from 31 to 72 C and pH`s from 8.0 to 10.5. At the higher temperatures and lower pH values, the results are described adequately by a shrinking core model that is first order with respect to hydroxyl concentration and particle surface area. Temperature effects were modeled with an Arrhenius equation, which indicates an activation energy of 35 kJ/mol. In separate experiments, the heat of reaction was determined to be 38.5 kJ/mol.

  5. Electrohydrolysis pretreatment of water hyacinth for enhanced hydrolysis. (United States)

    Barua, Visva Bharati; Raju, V Wilson; Lippold, Sophia; Kalamdhad, Ajay S


    This study investigates the use of electrohydrolysis pretreatment on water hyacinth to cut short the hydrolysis step and increase biogas production at the same time. Electrohydrolysis pretreatment of water hyacinth at 20V for 60min exhibited improved solubilisation (42.9%). Therefore, bio-chemical methane potential (BMP) test was carried out between water hyacinth pretreated at 20V for 60min and untreated water hyacinth. By the end of 30days, cumulative methane production of 2455±17mL CH4/g VS for electrohydrolysis pretreated substrate and 1936±27mL CH4/g VS for the untreated substrate was achieved. Compositional analysis and characterization study revealed the efficiency of electrohydrolysis pretreatment in melting the lignin and lowering the cellulose crystallinity of water hyacinth. Copyright © 2017 Elsevier Ltd. All rights reserved.

  6. Enhancing of sugar cane bagasse hydrolysis by Annulohypoxylon stygium glycohydrolases. (United States)

    Robl, Diogo; Costa, Patrícia dos Santos; Büchli, Fernanda; Lima, Deise Juliana da Silva; Delabona, Priscila da Silva; Squina, Fabio Marcio; Pimentel, Ida Chapaval; Padilla, Gabriel; Pradella, José Geraldo da Cruz


    The aim of this study was to develop a bioprocess for the production of β-glucosidase and pectinase from the fungus Annulohypoxylon stygium DR47. Media optimization and bioreactor cultivation using citrus bagasse and soybean bran were explored and revealed a maximum production of 6.26 U/mL of pectinase at pH 4.0 and 10.13 U/mL of β-glucosidase at pH 5.0. In addition, the enzymes extracts were able to replace partially Celluclast 1.5L in sugar cane bagasse hydrolysis. Proteomic analysis from A. stygium cultures revealed accessory enzymes, mainly belong to the families GH3 and GH54, that would support enhancement of commercial cocktail saccharification yields. This is the first report describing bioreactor optimization for enzyme production from A. stygium with a view for more efficient degradation of sugar cane bagasse. Copyright © 2014 Elsevier Ltd. All rights reserved.

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

    DEFF Research Database (Denmark)

    Tsai, Chien Tai

    , the cost of enzyme is still the bottle neck, re-using the enzyme is apossible way to reduce the input of enzyme in the process. In the point view of engineering, the prediction of enzymatic hydrolysis kinetics under different substrate loading, enzyme combination is usful for process design. Therefore....... Ionic liquid had been reported to be able to dissolve lignocellulose. However, as our knowledge, in all published researches, the concentration of lignocellulose in ionic liquid were low (5~10%). Besides, pretreatment time were long (from 1 hr to 1 day). Based on the hypothesis that the amount of ionic...... liquid and pretreatment time can be reduced, the influence of substrate concentration, pretreatment time and temperature were investigated and optimized. Pretreatment of barley straw by [EMIM]Ac, correlative models were constructed using 3 different pretreatment parameters (temperature, time...

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

    DEFF Research Database (Denmark)

    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 metal...... coordinating to the zinc ion. Potential proton shuttles from the second (unoccupied) metal-binding site (water, Asp, or Cys) are included in some calculations. The calculated reaction barrier for formation of the tetrahedral intermediate is 13 kcal/mol, close to what is observed experimentally for the rate......-limiting step. The barrier for the breakdown of the intermediate is low, 0-10 kcal/mol, if it is assisted by a water molecule or by a Cys or Asp model. Thus, the results indicate that proton transfer is not rate-limiting, and that any of the residues from the second metal site may function as proton shuttle...

  9. Dielectric barrier discharge plasma pretreatment on hydrolysis of microcrystalline cellulose (United States)

    Huang, Fangmin; Long, Zhouyang; Liu, Sa; Qin, Zhenglong


    Dielectric barrier discharge (DBD) plasma was used as a pretreatment method for downstream hydrolysis of microcrystalline cellulose (MCC). The degree of polymerization (DP) of MCC decreased after it was pretreated by DBD plasma under a carrier gas of air/argon. The effectiveness of depolymerization was found to be influenced by the crystallinity of MCC when under the pretreatment of DBD plasma. With the addition of tert-butyl alcohol in the treated MCC water suspension solution, depolymerization effectiveness of MCC was inhibited. When MCC was pretreated by DBD plasma for 30 min, the total reducing sugar concentration (TRSC) and liquefaction yield (LY) of pretreated-MCC (PMCC) increased by 82.98% and 34.18% respectively compared with those for raw MCC.

  10. IgG Glycan Hydrolysis Attenuates ANCA-Mediated Glomerulonephritis (United States)

    van Timmeren, Mirjan M.; van der Veen, Betty S.; Stegeman, Coen A.; Petersen, Arjen H.; Hellmark, Thomas; Collin, Mattias


    Anti-neutrophil cytoplasmic autoantibodies (ANCA) directed against myeloperoxidase (MPO) and proteinase 3 (Pr3) are considered pathogenic in ANCA-associated necrotizing and crescentic glomerulonephritis (NCGN) and vasculitis. Modulation of ANCA IgG glycosylation may potentially reduce its pathogenicity by abolishing Fc receptor–mediated activation of leukocytes and complement. Here, we investigated whether IgG hydrolysis by the bacterial enzyme endoglycosidase S (EndoS) attenuates ANCA-mediated NCGN. In vitro, treatment of ANCA IgG with EndoS significantly attenuated ANCA-mediated neutrophil activation without affecting antigen-binding capacity. In a mouse model of anti-MPO IgG/LPS-induced NCGN, we induced disease with either unmodified or EndoS-treated (deglycosylated) anti-MPO IgG. In separate experiments, we administered EndoS systemically after disease induction with unmodified anti-MPO IgG. Pretreatment of anti-MPO IgG with EndoS reduced hematuria, leukocyturia, and albuminuria and attenuated both neutrophil influx and formation of glomerular crescents. After inducing disease with unmodified anti-MPO IgG, systemic treatment with EndoS reduced albuminuria and glomerular crescent formation when initiated after 3 but not 24 hours. In conclusion, IgG glycan hydrolysis by EndoS attenuates ANCA-induced neutrophil activation in vitro and prevents induction of anti-MPO IgG/LPS-mediated NCGN in vivo. Systemic treatment with EndoS early after disease induction attenuates the development of disease. Thus, modulation of IgG glycosylation is a promising strategy to interfere with ANCA-mediated inflammatory processes. PMID:20448018

  11. Hydrolysis of phytic acid and its availability in rabbits. (United States)

    Marounek, M; Dusková, D; Skrivanová, V


    Twenty weaned rabbits were fed ad libitum two granulated feeds containing lucerne meal, barley, oats, wheat bran, oilseed meals and sugarbeet pulp in different proportions. Phytate P in these feeds represented 28.6 and 29.3 % of the total P. Digestibility trials were carried out in rabbits 7 and 10 weeks old. Digestibility of phytate P was 82.1 %, on average. Apparent digestibility of total P was 48.1 and 35.5 % in rabbits aged 7 and 10 weeks, respectively. Concentration of P in the faecal DM of these rabbits averaged 11.9 and 14.7 mg/g. Most of the faecal P was phosphates P (68.1 %). Proportion of phytate P in total faecal P was 9.0 %. Age effect on total P digestibility and faecal P concentration was significant (Pdigestive tract were based on the estimation of phytic acid hydrolysed during the first 2 h of incubation. The caecum contained 58.6 % of the phytase activity of the digestive tract. Corresponding relative values for the phytase activity in the stomach, small intestine and colon were 22.3, 7.7 and 11.4 %, respectively. In incubations of the caecal contents, phytic acid was hydrolysed more rapidly at pH 5-6 than in the neutral pH region. The hydrolysis was inhibited by Ca cations, and to a small extent also by phosphate anions. Commercial fungal phytase (Natupho) was highly active in incubations of the contents of the stomach at pH 1.9. It can be concluded that phytic acid is hydrolysed quite efficiently in the digestive tract of rabbits. This hydrolysis occurred mainly in the caecum. Absorption of soluble inorganic phosphates in the gut is incomplete.

  12. Kinetic study of enzymatic hydrolysis of potato starch

    Directory of Open Access Journals (Sweden)

    Óscar Fernando Castellanos Domínguez


    Full Text Available This article describes the kinetic study of potato starch enzymatic hydrolysis using soluble enzymes (Novo Nordisk. Different assays divided into four groups were used: reaction time (with which it was possible to reduce the 48-72 hour duration reported in the literature to 16 hours with comparable productivity levels; selecting the set of enzymes to be used (different types were evaluated - BAN and Termamyl as alfa-amylases during dextrinisation stage, and AMG, Promozyme and Fungamyl for sacarification reaction- identifying those presenting the best performance during hydrolysis.Reaction conditions were optimised for the process's two stages (destrinisation and sacarification. Enzyme dose, calcium cofactor concentration, pH, temperature and agitation speed were studied for the first stage. Enzyme ratio, pH and agitation speed were studied for sacarification; the latter parameter reported values having no antecedents in the literature (60 rpm and 30 rpm for first and second reactions, respectively. Michaelis Menten kinetics were calculated once conditions had been optimised, varying substrate from 10-50% P/V, obtaining km and Vmax kinetic parameters for each reaction. A kinetic model was found according to local working conditions which was able to explain potato starch conversion to glucose syrup, achieving 96 dextrose equivalents by the end of the reaction, being well within the maximum range reported in the literature (94-98.Laboratory equipment was constructed prior to carrying out assays which was able to reproduce and improve the conditions reported in the literature, making it a useful, reliable tool for use in assays returning good results.

  13. VLDL hydrolysis by hepatic lipase regulates PPARδ transcriptional responses.

    Directory of Open Access Journals (Sweden)

    Jonathan D Brown

    Full Text Available PPARs (α,γ,δ are a family of ligand-activated transcription factors that regulate energy balance, including lipid metabolism. Despite these critical functions, the integration between specific pathways of lipid metabolism and distinct PPAR responses remains obscure. Previous work has revealed that lipolytic pathways can activate PPARs. Whether hepatic lipase (HL, an enzyme that regulates VLDL and HDL catabolism, participates in PPAR responses is unknown.Using PPAR ligand binding domain transactivation assays, we found that HL interacted with triglyceride-rich VLDL (>HDL≫LDL, IDL to activate PPARδ preferentially over PPARα or PPARγ, an effect dependent on HL catalytic activity. In cell free ligand displacement assays, VLDL hydrolysis by HL activated PPARδ in a VLDL-concentration dependent manner. Extended further, VLDL stimulation of HL-expressing HUVECs and FAO hepatoma cells increased mRNA expression of canonical PPARδ target genes, including adipocyte differentiation related protein (ADRP, angiopoietin like protein 4 and pyruvate dehydrogenase kinase-4. HL/VLDL regulated ADRP through a PPRE in the promoter region of this gene. In vivo, adenoviral-mediated hepatic HL expression in C57BL/6 mice increased hepatic ADRP mRNA levels by 30%. In ob/ob mice, a model with higher triglycerides than C57BL/6 mice, HL overexpression increased ADRP expression by 70%, demonstrating the importance of triglyceride substrate for HL-mediated PPARδ activation. Global metabolite profiling identified HL/VLDL released fatty acids including oleic acid and palmitoleic acid that were capable of recapitulating PPARδ activation and ADRP gene regulation in vitro.These data define a novel pathway involving HL hydrolysis of VLDL that activates PPARδ through generation of specific monounsaturated fatty acids. These data also demonstrate how integrating cell biology with metabolomic approaches provides insight into specific lipid mediators and pathways of lipid

  14. Enzymatic lignocellulose hydrolysis: Improved cellulase productivity by insoluble solids recycling

    Directory of Open Access Journals (Sweden)

    Weiss Noah


    Full Text Available Abstract Background It is necessary to develop efficient methods to produce renewable fuels from lignocellulosic biomass. One of the main challenges to the industrialization of lignocellulose conversion processes is the large amount of cellulase enzymes used for the hydrolysis of cellulose. One method for decreasing the amount of enzyme used is to recycle the enzymes. In this study, the recycle of enzymes associated with the insoluble solid fraction after the enzymatic hydrolysis of cellulose was investigated for pretreated corn stover under a variety of recycling conditions. Results It was found that a significant amount of cellulase activity could be recovered by recycling the insoluble biomass fraction, and the enzyme dosage could be decreased by 30% to achieve the same glucose yields under the most favorable conditions. Enzyme productivity (g glucose produced/g enzyme applied increased between 30 and 50% by the recycling, depending on the reaction conditions. While increasing the amount of solids recycled increased process performance, the methods applicability was limited by its positive correlation with increasing total solids concentrations, reaction volumes, and lignin content of the insoluble residue. However, increasing amounts of lignin rich residue during the recycle did not negatively impact glucose yields. Conclusions To take advantage of this effect, the amount of solids recycled should be maximized, based on a given processes ability to deal with higher solids concentrations and volumes. Recycling of enzymes by recycling the insoluble solids fraction was thus shown to be an effective method to decrease enzyme usage, and research should be continued for its industrial application.


    Directory of Open Access Journals (Sweden)

    Dalton Longue Júnior


    Full Text Available The objective of this study was to assess the impact of removing hemicelluloses from chips of eucalyptus wood by self-hydrolysis treatment (H2O and on the subsequent ‘kraft’ process behavior and pulps bleachability and quality. The self-hydrolysis treatments were conducted at temperatures of 152°C (30, 45 and 60 minutes; 160°C (15, 30 and 45 minutes; and 170°C (5, 10, 15, 20 and 30 minutes; water: wood ratio of 4:1 m3/t. Normal chips (reference and self-hydrolyzed chips at 170°C during 5, 15 and 30 minutes were submitted to ‘kraft’ cooking up to kappa number 16 – 18 and the resulting pulp was bleached using the O/OD (EPO DD sequence. According to the results obtained, self-hydrolyzing the chips at 170°C for 30 minutes allowed the removal of up to 60% hemicelluloses. Cooking yield of the self-hydrolyzed chips for 30 minutes was around 6% smaller and pentosan content 88% lower than that of the regular chips. The efficiency of delignification with oxygen of the pulp derived from self-hydrolyzed chips for 30 minutes was of 75%, compared to 43.6% of the reference-pulp, and the bleaching cost using the O/OD (EPO DD sequence was US$ 7/adt per pulp, lower that that of the reference-pulp. The effluent originated from bleaching the pulp derived from the self-hydrolyzed chips presented lower values of COD (39.6%, color (21.3% and AOX (51.6%, compared to that of the reference-pulp.

  16. Optimization of enzymatic hydrolysis and fermentation conditions for improved bioethanol production from potato peel residues. (United States)

    Ben Taher, Imen; Fickers, Patrick; Chniti, Sofien; Hassouna, Mnasser


    The aim of this work was the optimization of the enzyme hydrolysis of potato peel residues (PPR) for bioethanol production. The process included a pretreatment step followed by an enzyme hydrolysis using crude enzyme system composed of cellulase, amylase and hemicellulase, produced by a mixed culture of Aspergillus niger and Trichoderma reesei. Hydrothermal, alkali and acid pretreatments were considered with regards to the enhancement of enzyme hydrolysis of potato peel residues. The obtained results showed that hydrothermal pretreatment lead to a higher enzyme hydrolysis yield compared to both acid and alkali pretreatments. Enzyme hydrolysis was also optimized for parameters such as temperature, pH, substrate loading and surfactant loading using a response surface methodology. Under optimized conditions, 77 g L -1 of reducing sugars were obtained. Yeast fermentation of the released reducing sugars led to an ethanol titer of 30 g L -1 after supplementation of the culture medium with ammonium sulfate. Moreover, a comparative study between acid and enzyme hydrolysis of potato peel residues was investigated. Results showed that enzyme hydrolysis offers higher yield of bioethanol production than acid hydrolysis. These results highlight the potential of second generation bioethanol production from potato peel residues treated with onsite produced hydrolytic enzymes. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:397-406, 2017. © 2017 American Institute of Chemical Engineers.

  17. Two-stage, dilute sulfuric acid hydrolysis of wood : an investigation of fundamentals (United States)

    John F. Harris; Andrew J. Baker; Anthony H. Conner; Thomas W. Jeffries; James L. Minor; Roger C. Pettersen; Ralph W. Scott; Edward L Springer; Theodore H. Wegner; John I. Zerbe


    This paper presents a fundamental analysis of the processing steps in the production of methanol from southern red oak (Quercus falcata Michx.) by two-stage dilute sulfuric acid hydrolysis. Data for hemicellulose and cellulose hydrolysis are correlated using models. This information is used to develop and evaluate a process design.

  18. Three-stage enzymatic hydrolysis of steam-exploded corn stover at high substrate concentration. (United States)

    Yang, Jing; Zhang, Xiaoping; Yong, Qiang; Yu, Shiyuan


    The feasibility of three-stage hydrolysis of steam-exploded corn stover at high-substrate concentration was investigated. When substrate concentration was 30% and enzyme loading was 15-30 FPU/g cellulose, three-stage (9+9+12 h) hydrolysis could reach a hydrolysis yield of 59.9-81.4% in 30 h. Compared with one-stage hydrolysis for 72 h, an increase of 34-37% in hydrolysis yield could be achieved. When steam-exploded corn stover was used as the substrate for enzyme synthesis and hydrolysis was conducted at a substrate concentration of 25% with an enzyme loading of 20 FPU/g cellulose, a hydrolysis yield of 85.1% was obtained, 19% higher than that the commercial cellulase could reach under the same conditions. The removal of end products was suggested to improve the adsorption of cellulase on the substrate and enhance the productivity of enzymatic hydrolysis. Copyright © 2010 Elsevier Ltd. All rights reserved.

  19. Kinetic study on the acid-catalyzed hydrolysis of cellulose to levulinic acid

    NARCIS (Netherlands)

    Girisuta, B.; Janssen, L. P. B. M.; Heeres, H. J.


    A variety of interesting bulk chemicals is accessible by the acid-catalyzed hydrolysis of cellulose. An interesting example is levulinic acid, a versatile precursor for fuel additives, polymers, and resins. A detailed kinetic study on the acid-catalyzed hydrolysis of cellulose to levulinic acid is

  20. The effect of high intensity mixing on the enzymatic hydrolysis of concentrated cellulose fiber suspensions (United States)

    Joseph R. Samaniuk; C. Tim Scott; Thatcher W. Root; Daniel J. Klingenberg


    Enzymatic hydrolysis of lignocellulosic biomass in a high shear environment was examined. The conversion of cellulose to glucose in samples mixed in a torque rheometer producing shear flows similar to those found in twin screw extruders was greater than that of unmixed samples. In addition, there is a synergistic effect of mixing and enzymatic hydrolysis; mixing...

  1. Processes and modeling of hydrolysis of particulate organic matter in aerobic wastewater tratment - A review

    DEFF Research Database (Denmark)

    Morgenroth, Eberhard Friedrich; Kommedal, Roald; Harremoës, Poul


    Carbon cycling and the availability of organic carbon for nutrient removal processes are in most wastewater treatment systems restricted by the rate of hydrolysis of slowly biodegradable (particulate) organic matter. To date, the mechanisms of hydrolysis are not well understood for complex...


    AbstractThe hydrolysis rates of mono-, di- and trihaloacetonitriles were studied in aqueous buffer solutions at different pH. The stability of haloacetonitriles decreases and the hydrolysis rate increases with increasing pH and number of halogen atoms in the molecule:...

  3. Production of sugar by hydrolysis of empty fruit bunches using palm ...

    African Journals Online (AJOL)

    The most challenging part in conversion of lignocellulosic materials to bioethanol is the hydrolysis process in order to obtain a reducing sugar. In this study, cellulase enzyme used for the hydrolysis was produced from palm oil mill effluent (POME), whose cost of production was considerably low as compared to commercial ...

  4. Kinetics of Strong Acid Hydrolysis of a Bleached Kraft Pulp for Producing Cellulose Nanocrystals (CNCs) (United States)

    Qianqian Wang; Xuebing Zhao; J.Y. Zhu


    Cellulose nanocrytals (CNCs) are predominantly produced using the traditional strong acid hydrolysis process. In most reported studies, the typical CNC yield is low (approximately 30%) despite process optimization. This study investigated the hydrolysis of a bleached kraft eucalyptus pulp using sulfuric acid between 50 and 64 wt % at temperatures of 35−80 °C...

  5. Simultaneous hydrolysis and hydrogenation of cellobiose to sorbitol in molten salt hydrate media

    NARCIS (Netherlands)

    Li, J.; Soares, H.S.M.P.; Moulijn, J.A.; Makkee, M.


    The hydrolysis and hydrogenation of cellobiose (4-O-b-D-glucopyranosyl-D-glucose) in ZnCl2_4H2O solvent was studied to optimize the conditions for conversion of lignocellulose (the most abundant renewable resource) into sorbitol (D-glucitol). Water at neutral pH does not allow hydrolysis of

  6. Effects of lignin-metal complexation on enzymatic hydrolysis of cellulose (United States)

    H. Liu; Junyong Zhu; S.Y. Fu


    This study investigated the inhibition of enzymatic hydrolysis by unbound lignin (soluble and insoluble) with or without the addition of metal compounds. Sulfonated, Organosolv, and Kraft lignin were added in aqueous enzyme-cellulose systems at different concentrations before hydrolysis. The measured substrate enzymatic digestibility (SED) of cellulose was decreased by...

  7. Hydrolysis of amphenicol and macrolide antibiotics: Chloramphenicol, florfenicol, spiramycin, and tylosin. (United States)

    Mitchell, Shannon M; Ullman, Jeffrey L; Teel, Amy L; Watts, Richard J


    Antibiotics that enter the environment can present human and ecological health risks. An understanding of antibiotic hydrolysis rates is important for predicting their environmental persistence as biologically active contaminants. In this study, hydrolysis rates and Arrhenius constants were determined as a function of pH and temperature for two amphenicol (chloramphenicol and florfenicol) and two macrolide (spiramycin and tylosin) antibiotics. Antibiotic hydrolysis rates in pH 4-9 buffer solutions at 25°C, 50°C, and 60°C were quantified, and degradation products were characterized. All of the antibiotics tested remained stable and exhibited no observable hydrolysis under ambient conditions typical of aquatic ecosystems. Acid- and base-catalyzed hydrolysis occurred at elevated temperatures (50-60°C), and hydrolysis rates increased considerably below pH 5 and above pH 8. Hydrolysis rates also increased approximately 1.5- to 2.9-fold for each 10°C increase in temperature. Based on the degradation product masses found, the functional groups that underwent hydrolysis were alkyl fluoride, amide, and cyclic ester (lactone) moieties; some of the resultant degradation products may remain bioactive, but to a lesser extent than the parent compounds. The results of this research demonstrate that amphenicol and macrolide antibiotics persist in aquatic systems under ambient temperature and pH conditions typical of natural waters. Thus, these antibiotics may present a risk in aquatic ecosystems depending on the concentration present. Copyright © 2015. Published by Elsevier Ltd.

  8. On the conflicting findings of Role of Cellulose-Crystallinity in Enzume Hydrolysis of Biomass (United States)

    Umesh Agarwal; Sally Ralph


    In the field of conversion of biomass to ethanol, an important area of research is the enzymatic hydrolysis of cellulose. Once cellulose is converted to glucose, it can be easily fermented to ethanol. As the cellulosic ethanol technology stands now, costly pretreatments and high dosages of cellulases are needed to achieve complete hydrolysis of the cellulose fraction...

  9. Hydrolysis mechanism of methyl parathion evidenced by Q-Exactive mass spectrometry. (United States)

    Liu, Yuan; Zhang, Caixiang; Liao, Xiaoping; Luo, Yinwen; Wu, Sisi; Wang, Jianwei


    Organophosphorus pesticides (OPPs), a kind of widely used pesticides, are currently attracting great attention due to their adverse effects on human central nervous systems, particularly in children. Although the hydrolysis behavior of OPPs has been studied well, its hydrolysis mechanism remained controversial, especially at various pH conditions, partly due to their relatively complex structures and abundant moieties that were prone to be attacked by nucleophiles. The Q-Exactive mass spectrometer, part of those hybrid high-resolution mass spectrometers (HRMS), was used to determine hydrolysis products of methyl parathion (MP), a kind of OPPs in situ buffer aqueous solution with pH ranging from 1 to 13 in this study. Most of the complex hydrolysis products of MP were identified due to the high sensitivity and accuracy of HRMS. The results demonstrated that the hydrolysis rate and pathway of MP were strong pH dependent. With the increase of pH, the hydrolysis rate of MP increased, and two different reaction mechanisms were identified: SN (2)@P pathway dominated the hydrolysis process at high pH (e.g., pH ≥ 11) while SN (2)@C was the main behavior at low pH (e.g., pH ≤ 9). This study helps understand the hydrolysis mechanism of OPPs at various pH and extends the use of Q-Exactive mass spectrometry in identifying organic pollutants and their degradation products in environmental matrices.

  10. Effects of Toasting Time on Digestive Hydrolysis of Soluble and Insoluble 00-Rapeseed Meal Proteins

    NARCIS (Netherlands)

    Salazar-Villanea, Sergio; Bruininx, Erik M.A.M.; Gruppen, Harry; Carré, Patrick; Quinsac, Alain; Poel, van der Thomas


    Thermal damage to proteins can reduce their nutritional value. The effects of toasting time on the kinetics of hydrolysis, the resulting molecular weight distribution of 00-rapeseed meal (RSM) and the soluble and insoluble protein fractions separated from the RSM were studied. Hydrolysis was

  11. Ultrasound mediated alkaline hydrolysis of methyl benzoate – reinvestigation with crucial parameters

    NARCIS (Netherlands)

    Sivakumara, Manickam; Senthilkumar, Paramasivam; Majumdara, Sukti; Pandit, Aniruddha B.


    In the present work hydrolysis of methyl benzoate was carried out using aqueous sodium hydroxide solution at room temperature in the presence of ultrasound since otherwise the same reaction takes place at relatively high temperature. Also, the above hydrolysis reaction was investigated at a

  12. Enzymatic hydrolysis as a means of expanding the cold gelation conditions of soy proteins

    NARCIS (Netherlands)

    Kuipers, B.J.H.; Koningsveld, van G.A.; Alting, A.C.; Driehuis, F.; Gruppen, H.; Voragen, A.G.J.


    Acid-induced cold gelation of soy protein hydrolysates was studied. Hydrolysates with degrees of hydrolysis (DH) of up to 10% were prepared by using subtilisin Carlsberg. The enzyme was inhibited to uncouple the hydrolysis from the subsequent gelation; the latter was induced by the addition of

  13. Pilot-scale base hydrolysis processing of HMX-based plastic-bonded explosives

    Energy Technology Data Exchange (ETDEWEB)

    Flesner, R.L.; Dell`Orco, P.C.; Spontarelli, T.; Bishop, R.L.; Skidmore, C.B.; Uher, K.; Kramer, J.F.


    Los Alamos National Laboratory has demonstrated that many energetic materials can be rendered non-energetic via reaction with sodium hydroxide or ammonia. This process is known as base hydrolysis. A pilot scale reactor has been developed to process up to 20 kg of plastic bonded explosive in a single batch operation. In this report, we discuss the design and operation of the pilot scale reactor for the processing of PBX 9404, a standard Department of Energy plastic bonded explosive containing HMX and nitrocellulose. Products from base hydrolysis, although non-energetic, still require additional processing before release to the environment. Decomposition products, destruction efficiencies, and rates of reaction for base hydrolysis will be presented. Hydrothermal processing, previously known as supercritical water oxidation, has been proposed for converting organic products from hydrolysis to carbon dioxide, nitrogen, and nitrous oxide. Base hydrolysis in combination with hydrothermal processing may yield a viable alternative to open burning/open detonation for destruction of many energetic materials.

  14. Pilot-scale base hydrolysis processing of HMX-based plastic-bonded explosives

    Energy Technology Data Exchange (ETDEWEB)

    Flesner, R.L.; Dell`orco, P.C.; Spontarelli, T.; Bishop, R.L.; Skidmore, C.; Uher, K.J.; Kramer, J.F.


    Los Alamos National Laboratory has demonstrated that many energetic materials can be rendered non-energetic via reaction with sodium hydroxide or ammonia. This process is known as base hydrolysis. A pilot scale reactor has been developed to process up to 20 kg of plastic bonded explosive in a single batch operation. In this report, we discuss the design and operation of the pilot scale reactor for the processing of PBX 9404, a standard Department of Energy plastic bonded explosive containing HMX and nitrocellulose. Products from base hydrolysis, although non-energetic, still require additional processing before release to the environment Decomposition products, destruction efficiencies, and rates of reaction for base hydrolysis will be presented. Hydrothermal processing, previously known as supercritical water oxidation, has been proposed for converting organic products from hydrolysis to carbon dioxide, nitrogen, and nitrous oxide. Base hydrolysis in combination with hydrothermal processing may yield a viable alternative to open burning/open detonation for destruction of many energetic materials.

  15. Effect of acid hydrolysis on starch structure and functionality: a review. (United States)

    Wang, Shujun; Copeland, Les


    Acid hydrolysis is an important chemical modification that can significantly change the structural and functional properties of starch without disrupting its granular morphology. A deep understanding of the effect of acid hydrolysis on starch structure and functionality is of great importance for starch scientific research and its industrial applications. During acid hydrolysis, amorphous regions are hydrolyzed preferentially, which enhances the crystallinity and double helical content of acid hydrolyzed starch. This review discusses current understanding of the effect of acid hydrolysis on starch structure and functionality. The effects of acid hydrolysis on amylose content, chain length distribution of amylopectin molecules, molecular and crystalline organization (including lamellar structure) and granular morphology are considered. Functional properties discussed include swelling power, gelatinization, retrogradation, pasting, gel texture, and in vitro enzyme digestibility. The paper also highlights some promising applications of acid hydrolyzed starch (starch nanocrystals) in the preparation of biodegradable nanocomposites, bio-hydrogen, and slowly digestible starch-based healthy foods.

  16. Product sampling during transient continuous countercurrent hydrolysis of canola oil and development of a kinetic model

    KAUST Repository

    Wang, Weicheng


    A chemical kinetic model has been developed for the transient stage of the continuous countercurrent hydrolysis of triglycerides to free fatty acids and glycerol. Departure functions and group contribution methods were applied to determine the equilibrium constants of the four reversible reactions in the kinetic model. Continuous countercurrent hydrolysis of canola oil in subcritical water was conducted experimentally in a lab-scale reactor over a range of temperatures and the concentrations of all neutral components were quantified. Several of the rate constants in the model were obtained by modeling this experimental data, with the remaining determined from calculated equilibrium constants. Some reactions not included in the present, or previous, hydrolysis modeling efforts were identified from glycerolysis kinetic studies and may explain the slight discrepancy between model and experiment. The rate constants determined in this paper indicate that diglycerides in the feedstock accelerate the transition from "emulsive hydrolysis" to "rapid hydrolysis". © 2013 Elsevier Ltd.

  17. CFD simulation of transient stage of continuous countercurrent hydrolysis of canola oil

    KAUST Repository

    Wang, Weicheng


    Computational Fluid Dynamic (CFD) modeling of a continuous countercurrent hydrolysis process was performed using ANSYS-CFX. The liquid properties and flow behavior such as density, specific heats, dynamic viscosity, thermal conductivity, and thermal expansivity as well as water solubility of the hydrolysis components triglyceride, diglyceride, monoglyceride, free fatty acid, and glycerol were calculated. Chemical kinetics for the hydrolysis reactions were simulated in this model by applying Arrhenius parameters. The simulation was based on actual experimental reaction conditions including temperature and water-to-oil ratio. The results not only have good agreement with experimental data but also show instantaneous distributions of concentrations of every component in hydrolysis reaction. This model provided visible insight into the continuous countercurrent hydrolysis process. © 2012 Elsevier Ltd.

  18. Process development of starch hydrolysis using mixing characteristics of Taylor vortices. (United States)

    Masuda, Hayato; Horie, Takafumi; Hubacz, Robert; Ohmura, Naoto; Shimoyamada, Makoto


    In food industries, enzymatic starch hydrolysis is an important process that consists of two steps: gelatinization and saccharification. One of the major difficulties in designing the starch hydrolysis process is the sharp change in its rheological properties. In this study, Taylor-Couette flow reactor was applied to continuous starch hydrolysis process. The concentration of reducing sugar produced via enzymatic hydrolysis was evaluated by varying operational variables: rotational speed of the inner cylinder, axial velocity (reaction time), amount of enzyme, and initial starch content in the slurry. When Taylor vortices were formed in the annular space, efficient hydrolysis occurred because Taylor vortices improved the mixing of gelatinized starch with enzyme. Furthermore, a modified inner cylinder was proposed, and its mixing performance was numerically investigated. The modified inner cylinder showed higher potential for enhanced mixing of gelatinized starch and the enzyme than the conventional cylinder.

  19. ATP hydrolysis assists phosphate release and promotes reaction ordering in F1-ATPase (United States)

    Li, Chun-Biu; Ueno, Hiroshi; Watanabe, Rikiya; Noji, Hiroyuki; Komatsuzaki, Tamiki


    F1-ATPase (F1) is a rotary motor protein that can efficiently convert chemical energy to mechanical work of rotation via fine coordination of its conformational motions and reaction sequences. Compared with reactant binding and product release, the ATP hydrolysis has relatively little contributions to the torque and chemical energy generation. To scrutinize possible roles of ATP hydrolysis, we investigate the detailed statistics of the catalytic dwells from high-speed single wild-type F1 observations. Here we report a small rotation during the catalytic dwell triggered by the ATP hydrolysis that is indiscernible in previous studies. Moreover, we find in freely rotating F1 that ATP hydrolysis is followed by the release of inorganic phosphate with low synthesis rates. Finally, we propose functional roles of the ATP hydrolysis as a key to kinetically unlock the subsequent phosphate release and promote the correct reaction ordering. PMID:26678797

  20. Interactions between single-chained ether phospholipids and sphingomyelin in mixed monolayers at the air/water interface-Grazing incidence X-ray diffraction and Brewster angle microscopy studies. (United States)

    Flasiński, Michał; Hąc-Wydro, Katarzyna; Wydro, Paweł; Broniatowski, Marcin; Dynarowicz-Łątka, Patrycja


    Single-chained ether phospholipids comprise a class of both natural (PAF, lyso-PAF) and synthetic (edelfosine, ED) compounds possessing confirmed extensive biological activities. Among them ED is known to exhibit antineoplastic properties, while PAF and its lyso-precursor are lipids implicated e.g. in the functioning of organism immune system. In our study the interactions of ED, PAF and lyso-PAF with sphingomyelin (SM) being one of the main lipid found in a high concentration in membrane microdomains, like lipid rafts, were investigated in mixed monolayers at the air/water interface. The traditional Langmuir methodology was complemented with modern physicochemical techniques: Grazing incidence X-ray diffraction and Brewster angle microscopy. The investigated compounds, i.e.: platelet activating factor (PAF), (lyso-PAF) and edelfosine were selected because of their highly different physiological properties despite very similar chemical structure and evidenced membrane activity. The obtained results demonstrate that all the investigated three single-chained phospholipids cause strong modification of the model membrane properties in a concentration dependent manner. It has been proved that there are significant differences regarding the influence of the single-chained lipids on model SM membrane--in the region of low concentration, edelfosine was found to be the most effective among all the investigated compounds. The collected data shed new light onto the membrane behavior of the investigated herein biochemically active compounds, which can be of help in understanding their different biological activity and designing of new, more biocompatible drugs. Copyright © 2013 Elsevier B.V. All rights reserved.

  1. The effect of pressure and temperature on aluminium hydrolysis: Implications to trace metal scavenging in natural waters

    Digital Repository Service at National Institute of Oceanography (India)

    DileepKumar, M.

    Removal of aluminium through precipitation/scavenging in natural waters was evaluated based on its hydrolysis at different temperatures and pressures. In general, pH for the occurrence of cation hydrolysis was lowered with hike in temperature which...

  2. Hydrolysis of whey lactose by immobilized β-Galactosidase

    Directory of Open Access Journals (Sweden)

    Marcela Panaro Mariotti


    Full Text Available Hydrolysis of whey lactose to glucose and galactose by immobilized galactosidase comes as an alternative to enlarge the possibilities of commercial use of this feedstock. To be applied at industrial scale, the process should be performed continuously .This work aimed to study the hydrolysis of whey lactose by an immobilized enzyme reactor. b-Galactosidase from Aspergillus oryzae was immobilized on silica and activity and stability were evaluated. The best immobilization results were attained by using glutaraldehyde as support's activator and enzyme stabilizer. The optimized enzyme proportion for immobilization was 15-20 mg g-1 of support. Treatments of whey were performed (microfiltration, thermal treatment and ultrafiltration, seeking the elimination of sludge, and the effects on operating the fixed bed reactor were evaluated. Ultrafiltration was the best treatment towards a proper substrate solution for feeding the reactor.A hidrólise de lactose de soro de leite, resultando em glicose e galactose, apresenta-se como uma alternativa para ampliar as possibilidades de uso comercial deste insumo. Para ser aplicado em escala industrial, o processo deve ser operado de modo contínuo. Reporta-se o estudo de um sistema objetivando hidrólise de lactose de soro de leite através de um reator com enzima imobilizada. b-Galactosidase de Aspergillus oryzae foi imobilizada em sílica, sendo avaliadas a estabilidade e atividade. Os melhores resultados de imobilização foram obtidos usando glutaraldeído como ativante do suporte e estabilizador da enzima. A proporção otimizada entre enzima e suporte foi 15-20 mg.g-1. Foram estudadas formas de tratamento do soro (microfiltração, tratamento térmico e ultrafiltração, objetivando eliminação de material suspenso, e avaliando os efeitos na operação de reator de leito fixo. A ultrafiltração foi o melhor tratamento, na busca de uma solução de substrato apropriada para o reator contínuo.

  3. Variation of structures of ingredients of desiccated coconut during hydrolysis by hydrochloric acid at low temperature

    Directory of Open Access Journals (Sweden)

    Jian XIONG


    Full Text Available Abstract Owing to the high content of lignocellulose, desiccated coconut become a healthy material for dietary fiber supplementation. In this study, the changes in solubility of the fibers of desiccated coconut were evaluated. The changes of the pHs and weight losses were studied. Furthermore, variations of the ingredient structures of desiccated coconut by hydrolysis by hydrochloric acid were characterized by Fourier transform infrared spectroscopy (FTIR, X-ray diffraction (XRD and scanning electron microscopy (SEM. After hydrolysis 30 s, the pHs of all systems increased, while six hours later, the pH of only system with initial pH = 1.00 decreased. The decline of pH only existed in hydrolysis systems with initial pH = 1.00, there is no relevant with the quantities of desiccated coconut. The lower initial pH of hydrolysis system was, the less the intrinsic viscosity of the desiccated coconut after hydrolysis was, the small the crystallinity was. After hydrolysis, the microstructure of the desiccated coconut become looser, and the secondary structure of the coconut protein became more stable and ordered. The results suggest that the hydrolysis of desiccated coconut mainly occurred in the branched chain and the non-crystalline region of lignocellulose, which transforms some insoluble dietary fiber into soluble dietary fiber. This improves the nutritional value of desiccated coconut.

  4. Hydrolysis of olive mill waste to enhance rhamnolipids and surfactin production. (United States)

    Moya Ramírez, Ignacio; Altmajer Vaz, Deisi; Banat, Ibrahim M; Marchant, Roger; Jurado Alameda, Encarnación; García Román, Miguel


    The aim of this work was to demonstrate the effectiveness of hydrolysis pretreatment of olive mill (OMW) waste before use as a carbon source in biosurfactant production by fermentation. Three hydrolysis methods were assessed: enzymatic hydrolysis, acid pretreatment plus enzymatic hydrolysis, and acid hydrolysis. Fermentation was carried out using two bacterial species: Pseudomonas aeruginosa and Bacillus subtilis. Our results showed that the enzymatic hydrolysis was the best pretreatment, yielding up to 29.5 and 13.7mg/L of rhamnolipids and surfactins respectively. Glucose did not show significant differences in comparison to enzymatically hydrolysed OMW. At the best conditions found rhamnolipids and surfactins reached concentrations of 299 and 26.5mg/L; values considerably higher than those obtained with non-hydrolysed OMW. In addition, enzymatic pretreatment seemed to partially reduce the inhibitory effects of OMW on surfactin production. Therefore, enzymatic hydrolysis proved to effectively increase the productivity of these biosurfactants using OMW as the sole carbon source. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. Effects of thermal hydrolysis temperature on physical characteristics of municipal sludge. (United States)

    Feng, Guohong; Guo, Yabing; Tan, Wei


    Effects of thermal hydrolysis temperature on the physical properties of municipal sludge was further studied by a series of experiments. There was a decrease in bound water content with an increase in hydrolysis temperature, while there was an increase in pH at temperatures below 120 °C, and a decrease at temperatures exceeding 120 °C. An analysis of settleability, centrifugation and vacuum filtration of the treated sludge indicated that the threshold temperature was 120 °C, which was the same as the temperature for the bound water content and particle size. In addition, raw sludge with a solids content of 100 g/L, exhibited significant non-Newtonian fluid characteristics. At thermal hydrolysis temperatures exceeding 120 °C, non-Newtonian fluid characteristics including liquid and solid characteristics were significantly weakened. The consistency index (k) decreased from 5.90 Pa·s to 0.068 Pa·s, while the flow index (n) increased from 0.31 to 0.74, suggesting that thermal hydrolysis sludge was much closer to Newtonian fluids compared to raw sludge. Modification of bound water content, particle size and viscosity with hydrolysis temperature, revealed the nature of improved dewaterability by thermal hydrolysis. The fractal dimension of the sludge floc increased from 2.74 to 2.90, meaning that the floc became more compact after thermal hydrolysis.

  6. Acid and enzymatic hydrolysis to recover reducing sugars from cassava bagasse: an economic study

    Directory of Open Access Journals (Sweden)

    Woiciechowski Adenise Lorenci


    Full Text Available The objective of this work was to study the acid and enzymatic hydrolysis of cassava bagasse for the recovery of reducing sugars and to establish the operational costs. A statistical program "Statistica", based on the surface response was used to optimize the recovery of reducing sugars in both the processes. The process economics was determined considering the values of reducing sugars obtained at laboratory scale, and the operations costs of a cylindrical reactor of 1500 L, with flat walls at the top and bottom. The reactor was operated with 150 kg of cassava bagasse and 1350 kg of water. The yield of the acid hydrolysis was 62.4 g of reducing sugars from 100 g of cassava bagasse containing 66% starch. It represented 94.5% of reducing sugar recovery. The yield of the enzymatic hydrolysis was 77.1 g of reducing sugars from 120 g of cassava bagasse, which represented 97.3% of reducing sugars recovery. Concerning to the time, a batch of acid hydrolysis required 10 minutes, plus the time to heat and cool the reactor, and a batch of the enzymatic hydrolysis needed 25 hours and 20 minutes, plus the time to heat and to cool the reactor. Thus, the acid hydrolysis of 150 kg of cassava bagasse required US$ 34.27, and the enzymatic hydrolysis of the same amount of cassava bagasse required US$ 2470.99.

  7. Effect of protonation on the mechanism of phosphate monoester hydrolysis and comparison with the hydrolysis of nucleoside triphosphate in biomolecular motors. (United States)

    Hassan, Hammad Ali; Rani, Sadaf; Fatima, Tabeer; Kiani, Farooq Ahmad; Fischer, Stefan


    Hydrolysis of phosphate groups is a crucial reaction in living cells. It involves the breaking of two strong bonds, i.e. the OaH bond of the attacking water molecule, and the POl bond of the substrate (Oa and Ol stand for attacking and leaving oxygen atoms). Mechanism of the hydrolysis reaction can proceed either by a concurrent or a sequential mechanism. In the concurrent mechanism, the breaking of OaH and POl bonds occurs simultaneously, whereas in the sequential mechanism, the OaH and POl bonds break at different stages of the reaction. To understand how protonation affects the mechanism of hydrolysis of phosphate monoester, we have studied the mechanism of hydrolysis of protonated and deprotonated phosphate monoester at M06-2X/6-311+G**//M06-2X/6-31+G*+ZPE level of theory (where ZPE stands for zero point energy). Our calculations show that in both protonated and deprotonated cases, the breaking of the water OaH bond occurs before the breaking of the POl bond. Because the two events are not separated by a stable intermediate, the mechanism can be categorized as semi-concurrent. The overall energy barrier is 41kcalmol(-1) in the unprotonated case. Most (5/6th) of this is due to the initial breaking of the water OaH bond. This component is lowered from 34 to 25kcalmol(-1) by adding one proton to the phosphate. The rest of the overall energy barrier comes from the subsequent breaking of the POl bond and is not sensitive to protonation. This is consistent with previous findings about the effect of triphosphate protonation on the hydrolysis, where the equivalent protonation (on the γ-phosphate) was seen to lower the barrier of breaking the water OaH bond and to have little effect on the POl bond breaking. Hydrolysis pathways of phosphate monoester with initial breaking of the POl bond could not be found here. This is because the leaving group in phosphate monoester cannot be protonated, unlike in triphosphate hydrolysis, where protonation of the β- and

  8. Standard Gibbs energy of metabolic reactions: II. Glucose-6-phosphatase reaction and ATP hydrolysis. (United States)

    Meurer, Florian; Do, Hoang Tam; Sadowski, Gabriele; Held, Christoph


    ATP (adenosine triphosphate) is a key reaction for metabolism. Tools from systems biology require standard reaction data in order to predict metabolic pathways accurately. However, literature values for standard Gibbs energy of ATP hydrolysis are highly uncertain and differ strongly from each other. Further, such data usually neglect the activity coefficients of reacting agents, and published data like this is apparent (condition-dependent) data instead of activity-based standard data. In this work a consistent value for the standard Gibbs energy of ATP hydrolysis was determined. The activity coefficients of reacting agents were modeled with electrolyte Perturbed-Chain Statistical Associating Fluid Theory (ePC-SAFT). The Gibbs energy of ATP hydrolysis was calculated by combining the standard Gibbs energies of hexokinase reaction and of glucose-6-phosphate hydrolysis. While the standard Gibbs energy of hexokinase reaction was taken from previous work, standard Gibbs energy of glucose-6-phosphate hydrolysis reaction was determined in this work. For this purpose, reaction equilibrium molalities of reacting agents were measured at pH7 and pH8 at 298.15K at varying initial reacting agent molalities. The corresponding activity coefficients at experimental equilibrium molalities were predicted with ePC-SAFT yielding the Gibbs energy of glucose-6-phosphate hydrolysis of -13.72±0.75kJ·mol(-1). Combined with the value for hexokinase, the standard Gibbs energy of ATP hydrolysis was finally found to be -31.55±1.27kJ·mol(-1). For both, ATP hydrolysis and glucose-6-phosphate hydrolysis, a good agreement with own and literature values were obtained when influences of pH, temperature, and activity coefficients were explicitly taken into account in order to calculate standard Gibbs energy at pH7, 298.15K and standard state. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Governing chemistry of cellulose hydrolysis in supercritical water. (United States)

    Cantero, Danilo A; Bermejo, M Dolores; Cocero, M José


    At extremely low reaction times (0.02 s), cellulose was hydrolyzed in supercritical water (T=400 °C and P=25 MPa) to obtain a sugar yield higher than 95 wt%, whereas the 5-hydroxymethylfurfural (5-HMF) yield was lower than 0.01 wt %. If the reaction time was increased to 1 s, the main product was glycolaldehyde (60 wt%). Independently of the reaction time, the yield of 5-HMF was always lower than 0.01 wt%. To evaluate the reaction mechanism of biomass hydrolysis in pressurized water, several parameters (temperature, pressure, reaction time, and reaction medium) were studied for different biomasses (cellulose, glucose, fructose, and wheat bran). It was found that the H(+) and OH(-) ion concentration in the reaction medium as a result of water dissociation is the determining factor in the selectivity. The reaction of glucose isomerization to fructose and the further dehydration to 5-HMF are highly dependent on the ion concentration. By an increase in the pOH/pH value, these reactions were minimized to allow control of 5-HMF production. Under these conditions, the retroaldol condensation pathway was enhanced, instead of the isomerization/dehydration pathway. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Sucrose Hydrolysis in a Bespoke Capillary Wall-Coated Microreactor

    Directory of Open Access Journals (Sweden)

    Filipe Carvalho


    Full Text Available Microscale technology has been increasingly used in chemical synthesis up to production scale, but in biocatalysis the implementation has been proceeding at a slower pace. In this work, the design of a low cost and versatile continuous flow enzyme microreactor is described that illustrates the potential of microfluidic reactors for both the development and characterization of biocatalytic processes. The core structure of the developed reactor consists of an array of capillaries with 450 μm of inner diameter with their inner surface functionalized with (3-aminopropyltriethoxysilane (APTES and glutaraldehyde where Saccharomyces cerevisiae invertase was covalently bound. The production of invert sugar syrup through enzymatic sucrose hydrolysis was used as model system. Once the microreactor assembly reproducibility and the immobilized enzyme behavior were established, the evaluation of the immobilized enzyme kinetic parameters was carried out at flow rates ranging from 20.8 to 219.0 μL·min−1 and substrate concentrations within 2.0%–10.0% (w/v. Despite the impact of immobilization on the kinetic parameters, viz. Km(app was increased two fold and Kcat showed a 14-fold decrease when compared to solution phase invertase, the immobilization proved highly robust. For a mean residence time of 48.8 min, full conversion of 5.0% (w/v sucrose was observed over 20 days.

  11. Characterization of acidic polysaccharides from the mollusks through acid hydrolysis. (United States)

    Cao, Jiuling; Wen, Chengrong; Lu, Jiaojiao; Teng, Nan; Song, Shuang; Zhu, Beiwei


    Uronic acid-containing polysaccharides (UACPs) including glycosaminoglycans (GAGs) exist widely in nature. Herein we propose an elegant methodology to identify UACPs by analyzing their disaccharides produced from the acid hydrolysis using HPLC-MS(n) upon 1-phenyl-3-methyl-5-pyrazolone (PMP) derivatization. Based on the optimization of experimental conditions by the single factor experiment and orthogonal test design, the combination of 1.3M TFA at 105°C for 3h is found to be the optimum. Subsequently, these conditions were applied to investigate the distribution of UACPs in 20 selected species of edible Bivalvia and Gastropoda. PMP-disaccharides derived from UACPs in mollusks were identified by comparing the retention time and mass spectra with those of the reference PMP-disaccharides from hyaluronic acid (HA), chondroitin sulfate (CS), heparin (HP), and AGSP with →4)-GlcA(1→2)-Man(1→ repeating units. The analysis reveals the prevalence of CS in the shellfishes as well as the HP, and existence of three non-GAG UACPs in 7 mollusks. Copyright © 2015 Elsevier Ltd. All rights reserved.

  12. Hydrolysis of sweet blue lupin hull using subcritical water technology. (United States)

    Ciftci, Deniz; Saldaña, Marleny D A


    Hydrolysis of sweet blue lupin hulls was conducted in this study using subcritical water technology. Effects of process parameters, such as pressure (50-200 bar), temperature (160-220°C), flow rate (2-10 mL/min), and pH (2-12), were studied to optimize maximum hemicellulose sugars recovery in the extracts. Extracts were analyzed for total hemicellulose sugars, phenolics and organic carbon contents and solid residues left after treatments were also characterized. Temperature, flow rate, and pH had a significant effect on hemicellulose sugar removal; however, the effect of pressure was not significant. The highest yield of hemicellulose sugars in the extracts (85.5%) was found at 180°C, 50 bar, 5 mL/min and pH 6.2. The thermal stability of the solid residue obtained at optimum conditions improved after treatment and the crystallinity index increased from 11.5% to 58.6%. The results suggest that subcritical water treatment is a promising technology for hemicellulose sugars removal from biomass. Copyright © 2015 Elsevier Ltd. All rights reserved.

  13. Pequi cake composition, hydrolysis and fermentation to bioethanol

    Directory of Open Access Journals (Sweden)

    A. L. Macedo


    Full Text Available Pequizeiro (Caryocar brasiliense Camb fruits have been evaluated as a potential raw material for the newly established biodiesel industry. This scenario demands applications using the solid co-product derived from the extraction of pequi oil, called cake or meal. This study analyses the acid hydrolysis of carbohydrates present in the pequi meal in order to obtain fermentable sugars and evaluates their conversion to bioethanol. There was 27% starch in the pequi meal. The use of a CCRD experimental design type to study the acid saccharification of pequi meal results in 61.6% conversion of its starch content to reducing sugars. Positive and significant linear effects were observed for H2SO4 concentration and temperature factors, while the quadratic effect of H2SO4 concentration and the linear effect of solid-liquid ratio were negative. Even, with non-optimized fermentative condition using 1% of dried baker's yeast in conical flasks, it was possible to obtain a value equivalent to 53 L of ethanol per ton of hydrolyzed pequi meal.

  14. Modeling residence-time distribution in horizontal screw hydrolysis reactors

    Energy Technology Data Exchange (ETDEWEB)

    Sievers, David A.; Stickel, Jonathan J.


    The dilute-acid thermochemical hydrolysis step used in the production of liquid fuels from lignocellulosic biomass requires precise residence-time control to achieve high monomeric sugar yields. Difficulty has been encountered reproducing residence times and yields when small batch reaction conditions are scaled up to larger pilot-scale horizontal auger-tube type continuous reactors. A commonly used naive model estimated residence times of 6.2-16.7 min, but measured mean times were actually 1.4-2.2 the estimates. This study investigated how reactor residence-time distribution (RTD) is affected by reactor characteristics and operational conditions, and developed a method to accurately predict the RTD based on key parameters. Screw speed, reactor physical dimensions, throughput rate, and process material density were identified as major factors affecting both the mean and standard deviation of RTDs. The general shape of RTDs was consistent with a constant value determined for skewness. The Peclet number quantified reactor plug-flow performance, which ranged between 20 and 357.

  15. Radiolysis and hydrolysis of magnetically assisted chemical separation particles

    Energy Technology Data Exchange (ETDEWEB)

    Buchholz, B.A.; Nunez, L.; Vandegrift, G.F.


    The magnetically assisted chemical separation (MACS) process is designed to separate transuranic (TRU) elements out of high-level waste (HLW) or TRU waste. Magnetic microparticles (1--25 {mu}m) were coated with octyl (phenyl)N,N-diisobutylcarbamoylmethylphosphine oxide (CMPO) dissolved in tributyl phosphate (TBP) and tested for removing TRU elements from acidic nitrate solutions. The particles were contacted with nitric acid solutions and Hanford plutonium finishing plant (PFP) simulant, irradiated with a high intensity {sup 60}Co {gamma}-ray source, and evaluated for effectiveness in removing TRU elements from 2m HNO{sub 3} solutions. The resistance of the coatings and magnetic cores to radiolytic damage and hydrolytic degradation was investigated by irradiating samples of particles suspended in a variety of solutions with doses of up to 5 Mrad. Transmission electron microscopy (TEM), magnetic susceptibility measurements, and physical observations of the particles and suspension solutions were used to assess physical changes to the particles. Processes that affect the surface of the particles dramatically alter the binding sites for TRU in solution. Hydrolysis played a larger role than radiolysis in the degradation of the extraction capacity of the particles.

  16. Green fuel production from saffron waste by dilute acid hydrolysis

    Directory of Open Access Journals (Sweden)

    Ali ArastehNodeh


    Full Text Available In this study, the dilute acid Hydrolysis of saffron waste as a lignocellulose biomass for fermentable sugar production as a green fuel is investigated. Saffron plant waste was collected, washed with distilled water, dried by being exposed to air and sunlight, milled by a disk mill up to 50 micrometers and kept in plastic at room temperature. The prepared biomass was heated in a specified acidic solution for a predetermined time period in the autoclave. The influences of 3 major parameters including temperature, acid concentration and time on the produced fermentable sugar is investigated by the response surface method. According to previous studies, the levels for pretreatment were: temperature (116, 124, 132, and 140oC, process time (20, 30, 40, and 50 minute and sulfuric acid concentration (0, 1, 2, and 3%. To determine the type and quantity of products, the filtered solution was analyzed by HPLC and the results (glucose, xylose and furfural were modeled by a quadratic equation. This model showed that glucose concentration was influenced by the quadratic power of acid concentration and time while xylose concentration was influenced by time and furfural depends on time alone. It may be concluded that high acid concentration, low temperature and medium time could produce maximum products (glucose and xylose with minimum inhibitor (furfural.

  17. Chemical and Enzymatic Hydrolysis of Polyurethane/Polylactide Blends

    Directory of Open Access Journals (Sweden)

    Joanna Brzeska


    Full Text Available Polyether-esterurethanes containing synthetic poly[(R,S-3-hydroxybutyrate] (R,S-PHB and polyoxytetramethylenediol in soft segments and polyesterurethanes with poly(ε-caprolactone and poly[(R,S-3-hydroxybutyrate] were blended with poly([D,L]-lactide (PLA. The products were tested in terms of their oil and water absorption. Oil sorption tests of polyether-esterurethane revealed their higher response in comparison to polyesterurethanes. Blending of polyether-esterurethanes with PLA caused the increase of oil sorption. The highest water sorption was observed for blends of polyether-esterurethane, obtained with 10% of R,S-PHB in soft segments. The samples mass of polyurethanes and their blends were almost not changed after incubation in phosphate buffer and trypsin and lipase solutions. Nevertheless the molecular weight of polymers was significantly reduced after degradation. It was especially visible in case of incubation of samples in phosphate buffer what suggested the chemical hydrolysis of polymer chains. The changes of surface of polyurethanes and their blends, after incubation in both enzymatic solutions, indicated on enzymatic degradation, which had been started despite the lack of mass lost. Polyurethanes and their blends, contained more R,S-PHB in soft segments, were degraded faster.

  18. The computational analysis and modelling of substitution effects on hydrolysis of formanilides in acidic aqueous solutions (United States)

    Lukeš, Vladimír; Škorňa, Peter; Michalík, Martin; Klein, Erik


    Various para, meta and ortho substituted formanilides have been theoretically studied. For trans and cis-isomers of non-substituted formanilide, the calculated B3LYP vibration normal modes were analyzed. Substituent effect on the selected normal modes was described and the comparison with the available experimental data is presented. The calculated B3LYP proton affinities were correlated with Hammett constants, Fujita-Nishioka equation and the rate constants of the hydrolysis in 1 M HCl. Found linear dependences allow predictions of dissociation constants (pKBH+) and hydrolysis rate constants. Obtained results indicate that protonation of amide group may represent the rate determining step of acid catalyzed hydrolysis.

  19. Pre-steady-state Kinetics for Hydrolysis of Insoluble Cellulose by Cellobiohydrolase Cel7A

    DEFF Research Database (Denmark)

    Cruys-Bagger, Nicolaj; Olsen, Jens Elmerdahl; Præstgaard, Eigil


    complex, processive hydrolysis, and dissociation, respectively. These kinetic parameters elucidate limiting factors in the cellulolytic process. We concluded, for example, that Cel7A cleaves about four glycosidic bonds/s during processive hydrolysis. However, the results suggest that stalling...... for the exo-acting cellulase Cel7A using amperometric biosensors and an explicit model for processive hydrolysis of cellulose. This analysis allows the identification of a pseudo-steady-state period and quantification of a processivity number as well as rate constants for the formation of a threaded enzyme...

  20. Effects of copper source and concentration on in vitro phytate phosphorus hydrolysis by phytase. (United States)

    Pang, Yanfang; Applegate, Todd J


    Five copper (Cu) sources were studied at pH 2.5, 5.5, and 6.5 to determine how Cu affects phytate phosphorus (PP) hydrolysis by phytase at concentrations up to 500 mg/kg diet (60 min, 40-41 degrees C). Subsequently, Cu solubility with and without sodium phytate was measured. Adding Cu inhibited PP hydrolysis at pH 5.5 and pH 6.5 (P copper chloride and copper lysinate inhibited PP hydrolysis much less than copper sulfate pentahydrate, copper chloride, and copper citrate (P copper-phytin complexes.

  1. Hydrolysis rate constants at 10-25 °C can be more than doubled by a short anaerobic pre-hydrolysis at 35 °C. (United States)

    Zhang, L; Gao, R; Naka, A; Hendrickx, T L G; Rijnaarts, H H M; Zeeman, G


    Hydrolysis is the first step of the anaerobic digestion of complex wastewater and considered as the rate limiting step especially at low temperature. Low temperature (10-25 °C) hydrolysis was investigated with and without application of a short pre-hydrolysis at 35 °C. Batch experiments were executed using cellulose and tributyrin as model substrates for carbohydrates and lipids. The results showed that the low temperature anaerobic hydrolysis rate constants increased by a factor of 1.5-10, when the short anaerobic pre-hydrolysis at 35 °C was applied. After the pre-hydrolysis phase at 35 °C and decreasing the temperature, no lag phase was observed in any case. Without the pre-hydrolysis, the lag phase for cellulose hydrolysis at 35-10 °C was 4-30 days. Tributyrin hydrolysis showed no lag phase at any temperature. The hydrolysis efficiency of cellulose increased from 40 to 62%, and from 9.6 to 40% after 9.1 days at 15 and 10 °C, respectively, when the pre-hydrolysis at 35 °C was applied. The hydrolysis efficiency of tributyrin at low temperatures with the pre-hydrolysis at 35 °C was similar to those without the pre-hydrolysis. The hydrolytic activity of the supernatant collected from the digestate after batch digestion of cellulose and tributyrin at 35 °C was higher than that of the supernatants collected from the low temperature (≤25 °C) digestates. Copyright © 2016 Elsevier Ltd. All rights reserved.

  2. The Acid Hydrolysis Mechanism of Acetals Catalyzed by a Supramolecular Assembly in Basic Solution

    Energy Technology Data Exchange (ETDEWEB)

    Pluth, Michael D.; Bergman, Robert G.; Raymond, Kenneth N.


    A self-assembled supramolecular host catalyzes the hydrolysis of acetals in basic aqueous solution. The mechanism of hydrolysis is consistent with the Michaelis-Menten kinetic model. Further investigation of the rate limiting step of the reaction revealed a negative entropy of activation ({Delta}S{double_dagger} = -9 cal mol{sup -1}K{sup -1}) and an inverse solvent isotope effect (k(H{sub 2}O)/k(D{sub 2}O) = 0.62). These data suggest that the mechanism of hydrolysis that takes place inside the assembly proceeds through an A-2 mechanism, in contrast to the A-1 mechanism operating in the uncatalyzed reaction. Comparison of the rates of acetal hydrolysis in the assembly with the rate of the reaction of unencapsulated substrates reveals rate accelerations of up to 980 over the background reaction for the substrate diethoxymethane.

  3. Comparison of aqueous ammonia and dilute acid pretreatment of bamboo fractions: Structure properties and enzymatic hydrolysis. (United States)

    Xin, Donglin; Yang, Zhong; Liu, Feng; Xu, Xueru; Zhang, Junhua


    The effect of two pretreatments methods, aqueous ammonia (SAA) and dilute acid (DA), on the chemical compositions, cellulose crystallinity, morphologic change, and enzymatic hydrolysis of bamboo fractions (bamboo yellow, timber, green, and knot) was compared. Bamboo fractions with SAA pretreatment had better hydrolysability than those with DA pretreatment. High crystallinity index resulted in low hydrolysis yield in the conversion of SAA pretreated bamboo fractions, not DA pretreated fractions. The increase of cellulase loading had modestly positive effect in the hydrolysis of both SAA and DA pretreated bamboo fractions, while supplement of xylanase significantly increased the hydrolysis of the pretreated bamboo fractions, especially after SAA pretreatment. The results indicated that SAA pretreatment was more effective than DA pretreatment in conversion of bamboo fractions, and supplementation of xylanase was necessary in effective conversion of the SAA pretreated fractions into fermentable sugars. Copyright © 2014 Elsevier Ltd. All rights reserved.

  4. Dynamic Simulation, Sensitivity and Uncertainty Analysis of a Demonstration Scale Lignocellulosic Enzymatic Hydrolysis Process

    DEFF Research Database (Denmark)

    Prunescu, Remus Mihail; Sin, Gürkan


    This study presents the uncertainty and sensitivity analysis of a lignocellulosic enzymatic hydrolysis model considering both model and feed parameters as sources of uncertainty. The dynamic model is parametrized for accommodating various types of biomass, and different enzymatic complexes...

  5. Prediction of Hydrolysis Products of Organic Chemicals under Environmental pH Conditions (United States)

    Cheminformatics-based software tools can predict the molecular structure of transformation products using a library of transformation reaction schemes. This paper presents the development of such a library for abiotic hydrolysis of organic chemicals under environmentally relevant...

  6. Hydrothermal pretreatment and enzymatic hydrolysis of mixed green and woody lignocellulosics from arid regions

    DEFF Research Database (Denmark)

    Ashraf, Muhammad Tahir; Thomsen, Mette Hedegaard; Schmidt, Jens Ejbye


    Utilization of multi-specie feedstocks is imperative for application of lignocellulosic biorefineries in arid regions. Different lignocellulosic residues vary in composition and anatomical features. Pretreatment and enzymatic hydrolysis are two processes at the front end of any lignocellulosics b...

  7. Comparison of dilute mineral and organic acid pretreatment for enzymatic hydrolysis of wheat straw

    NARCIS (Netherlands)

    Kootstra, A.M.J.; Beeftink, H.H.; Scott, E.L.; Sanders, J.P.M.


    The efficiencies of fumaric, maleic, and sulfuric acid in wheat straw pretreatment were compared. As a measure for pretreatment efficiency, enzymatic digestibility of the lignocellulose was determined. Monomeric glucose and xylose concentrations were measured after subsequent enzymatic hydrolysis,

  8. Recovery and reuse of cellulase catalyst in an exzymatic cellulose hydrolysis process (United States)

    Woodward, Jonathan


    A process for recovering cellulase from the hydrolysis of cellulose, and reusing it in subsequent hydrolyois procedures. The process utilizes a commercial adsorbent that efficiently removes cellulase from reaction products which can be easily removed by simple decantation.

  9. Biological pretreatment of rubberwood with Ceriporiopsis subvermispora for enzymatic hydrolysis and bioethanol production

    National Research Council Canada - National Science Library

    Nazarpour, Forough; Abdullah, Dzulkefly Kuang; Abdullah, Norhafizah; Motedayen, Nazila; Zamiri, Reza


    Rubberwood (Hevea brasiliensis), a potential raw material for bioethanol production due to its high cellulose content, was used as a novel feedstock for enzymatic hydrolysis and bioethanol production using biological pretreatment...

  10. Alpha-tricalcium phosphate hydrolysis to octacalcium phosphate: effect of sodium polyacrylate. (United States)

    Bigi, A; Boanini, E; Botter, R; Panzavolta, S; Rubini, K


    Alpha-Tricalcium phosphate (alpha-TCP) hydrolysis into octacalcium phosphate (OCP) has been investigated in phosphoric acid solution at different concentrations of sodium polyacrylate (NaPA). The hydrolysis process has been followed by powder X-ray diffraction, infrared absorption and scanning electron microscopy analyses. In the absence of the polyelectrolyte, alpha-TCP undergoes a complete transformation into OCP in 24 h. The presence of polyacrylate in solution inhibits the hydrolysis so that a NaPA concentration of 0.5 microm is sufficient to lengthen the time required to complete the hydrolysis to 4 days. The variation of Ca2+ concentration in the soaking solution suggests that the transformation occurs through alpha-TCP dissolution followed by OCP precipitation. The delayed OCP nucleation and growth in the presence of polyacrylate implies a preferential adsorption of the polyelectrolyte on the growing OCP crystals, which induces an anisotropic reduction of the coherence lengths of the perfect crystalline domains.

  11. Improving enzymatic hydrolysis of industrial hemp (Cannabis sativa L.) by electron beam irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Shin, Soo-Jeong [Chungbuk National University, Cheongju, Chungbuk 361-763 (Korea, Republic of); Sung, Yong Joo [KT and G Central Research Institute, 302 Shinseong-Dong, Yuseong-Gu, Daejeon 305-805 (Korea, Republic of)], E-mail:


    The electron beam irradiation was applied as a pretreatment of the enzymatic hydrolysis of hemp biomass with doses of 150, 300 and 450 kGy. The higher irradiation dose resulted in the more extraction with hot-water extraction or 1% sodium hydroxide solution extraction. The higher solubility of the treated sample was originated from the chains scission during irradiation, which was indirectly demonstrated by the increase of carbonyl groups as shown in diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS) spectra. The changes in the micro-structure of hemp resulted in the better response to enzymatic hydrolysis with commercial cellulases (Celluclast 1.5L and Novozym 342). The improvement in enzymatic hydrolysis by the irradiation was more evident in the hydrolysis of the xylan than in that of the cellulose.

  12. Hydrolysis of carotenoid esters from Tagetes erecta by the action of lipases from Yarrowia lipolytica

    National Research Council Canada - National Science Library

    Abdala, Abraham Figueiras; Gallardo, Alfonso Pérez; Olvera, Lorenzo Guevara; Silva, Eleazar Máximo Escamilla


    The present study was conducted to evaluate the feasibility of enzymatic hydrolysis of carotenoid esters from Tagetes erecta using lipases from the yeast of Yarrowia lipolytica, with the aim of obtaining free lutein...

  13. Hydrothermal pretreatment and enzymatic hydrolysis of mixed green and woody lignocellulosics from arid regions. (United States)

    Ashraf, Muhammad Tahir; Thomsen, Mette Hedegaard; Schmidt, Jens Ejbye


    Utilization of multi-specie feedstocks is imperative for application of lignocellulosic biorefineries in arid regions. Different lignocellulosic residues vary in composition and anatomical features. Pretreatment and enzymatic hydrolysis are two processes at the front end of any lignocellulosics biorefinery applying biochemical pathway, and have to efficiently deal with the variance in the feedstock composition and properties. However, there is limited knowledge about effect of mixing different lignocellulosics on pretreatment and enzymatic hydrolysis yields. In this study effect of mixing on the yields from hydrothermal pretreatment and enzymatic hydrolysis was analyzed by mixing three different lignocellulosic residues - Bermuda grass, Jasmine hedges, and date palm fronds. Results showed that the individual and the mixed lignocellulosics gave same yields when treated under similar conditions of hydrothermal pretreatment and enzymatic hydrolysis. It indicates that this mixture can be a suitable feedstock for lignocellulosic biorefinery. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. Product inhibition of enzymatic hydrolysis of cellulose: are we running the reactions all wrong?

    DEFF Research Database (Denmark)

    Meyer, Anne S.


    include high substrate conversion (maximal yields), maximal enzyme efficiency, maximal volumetric reactor productivity, minimal equipment investment, minimal size, and short reaction time. The classic batch type STR reactions used for enzymatic cellulose hydrolysis prevent these goals to be fulfilled...

  15. Crystallization study and comparative in vitro-in vivo hydrolysis of PLA reinforcement ligament

    National Research Council Canada - National Science Library

    Beslikas, Theodore; Gigis, Ioannis; Goulios, Vasilios; Christoforides, John; Papageorgiou, George Z; Bikiaris, Dimitrios N


    In the present work, the crystallization behavior and in vitro-in vivo hydrolysis rates of PLA absorbable reinforcement ligaments used in orthopaedics for the repair and reinforcement of articulation...

  16. Effect of acid hydrolysis of Garcinia kola (bitter kola) pulp waste on ...

    African Journals Online (AJOL)



    -cellulase and β-glucosidase compared with maize and rice husks. Key words: Bitter kola pulp wastes, CM-cellulase, β-glucosidase, Aspergillus niger, acid hydrolysis. INTRODUCTION. Cellulose is the most abundant organic ...

  17. Chloramine-T-mediated chemoselective hydrolysis of thioglycosides into glycosyl hemiacetals under neutral conditions. (United States)

    Misra, Anup Kumar; Agnihotri, Geetanjali


    A metal-free, mild, efficient and chemoselective hydrolysis of several thioalkylglycosides (1) into their corresponding 1-hydroxy sugars (2) using sodium N-chloro-p-toluenesulfonamide trihydrate (chloramine-T) without affecting other functional groups is reported.

  18. Effect of enzymatic hydrolysis on native starch granule structure. (United States)

    Blazek, Jaroslav; Gilbert, Elliot Paul


    Enzymatic digestion of six starches of different botanical origin was studied in real time by in situ time-resolved small-angle neutron scattering (SANS) and complemented by the analysis of native and digested material by X-ray diffraction, differential scanning calorimetry, small-angle X-ray scattering, and scanning electron microscopy with the aim of following changes in starch granule nanostructure during enzymatic digestion. This range of techniques enables coverage over five orders of length-scale, as is necessary for this hierarchically structured material. Starches studied varied in their digestibility and displayed structural differences in the course of enzymatic digestion. The use of time-resolved SANS showed that solvent-drying of digested residues does not induce any structural artifacts on the length scale followed by small-angle scattering. In the course of digestion, the lamellar peak intensity gradually decreased and low-q scattering increased. These trends were more substantial for A-type than for B-type starches. These observations were explained by preferential digestion of the amorphous growth rings. Hydrolysis of the semicrystalline growth rings was explained on the basis of a liquid-crystalline model for starch considering differences between A-type and B-type starches in the length and rigidity of amylopectin spacers and branches. As evidenced by differing morphologies of enzymatic attack among varieties, the existence of granular pores and channels and physical penetrability of the amorphous growth ring affect the accessibility of the enzyme to the substrate. The combined effects of the granule microstructure and the nanostructure of the growth rings influence the opportunity of the enzyme to access its substrate; as a consequence, these structures determine the enzymatic digestibility of granular starches more than the absolute physical densities of the amorphous growth rings and amorphous and crystalline regions of the semicrystalline

  19. Effect of salting on back fat hydrolysis and oxidation (United States)

    Tunieva, E. К; Nasonova, V. V.; Stanovova, I. A.; Spiridonov, К I.; Kurzova, A. A.


    Technological factors significantly affect the rate of hydrolytic and oxidative changes in fat. The aim of the research was to study the effect of sodium chloride on hydrolysis and oxidation of fat raw material, including the impact of thermal treatment. Back fat was minced, sodium chloride was added (in amounts of 0.0, 2.0, 3.5 or 5.0%), then it was thermally treated or not. Determination of the acid value (AV) was carried out by titration with aqueous potassium hydroxide of free fatty acids in the ether-alcohol solution of back fat; the peroxide value (PV) was based on oxidation of iodhydric acid with peroxides contained in fat followed by titration of released iodine with sodium thiosulphate. The thiobarbituric acid value (TBAV) was determined by the development of stained substances due to interaction of fat oxidation products with 2-thiobarbituric acid and measurement of color intensity using a spectrophotometer. Adding 5.0% sodium chloride to back fat led to a 30.1% decrease in AV. Addition of 2.0% sodium chloride inhibited the development of the oxidation products and led to a 17% decrease in the PV and to a 25% decrease in TBAV (p0.05). The results obtained showed an ambiguous effect of sodium chloride on the processes of fat oxidation, depending on dosage and the use of thermal treatment, justifying the necessity to develop approaches that allow reduction of the sodium chloride content in meat products that are not subjected to thermal treatment.

  20. Studies on the Mechanism of Ring Hydrolysis in Phenylacetate Degradation (United States)

    Teufel, Robin; Gantert, Carla; Voss, Michaela; Eisenreich, Wolfgang; Haehnel, Wolfgang; Fuchs, Georg


    The widespread, long sought-after bacterial aerobic phenylalanine/phenylacetate catabolic pathway has recently been elucidated. It proceeds via coenzyme A (CoA) thioesters and involves the epoxidation of the aromatic ring of phenylacetyl-CoA, subsequent isomerization to an uncommon seven-membered C-O-heterocycle (oxepin-CoA), and non-oxygenolytic ring cleavage. Here we characterize the hydrolytic oxepin-CoA ring cleavage catalyzed by the bifunctional fusion protein PaaZ. The enzyme consists of a C-terminal (R)-specific enoyl-CoA hydratase domain (formerly MaoC) that cleaves the ring and produces a highly reactive aldehyde and an N-terminal NADP+-dependent aldehyde dehydrogenase domain that oxidizes the aldehyde to 3-oxo-5,6-dehydrosuberyl-CoA. In many phenylacetate-utilizing bacteria, the genes for the pathway exist in a cluster that contains an NAD+-dependent aldehyde dehydrogenase in place of PaaZ, whereas the aldehyde-producing hydratase is encoded outside of the cluster. If not oxidized immediately, the reactive aldehyde condenses intramolecularly to a stable cyclic derivative that is largely prevented by PaaZ fusion in vivo. Interestingly, the derivative likely serves as the starting material for the synthesis of antibiotics (e.g. tropodithietic acid) and other tropone/tropolone related compounds as well as for ω-cycloheptyl fatty acids. Apparently, bacteria made a virtue out of the necessity of disposing the dead-end product with ring hydrolysis as a metabolic branching point. PMID:21296885

  1. Hydrolysis of pyridoxal-5'-phosphate in plasma in conditions with raised alkaline phosphate.


    Anderson, B B; O'Brien, H; Griffin, G.E.; Mollin, D L


    Hydrolysis of pyridoxal phosphate in plasma was demonstrated in patients with liver disease and other conditions with raised alkaline phosphatase, and this usually closely paralleled the alkaline phosphatase level, whether of liver or bone origin. The endogenous plasma pyridoxal phosphate was inversely related to the alkaline phosphatase, and plasma hydrolysis of pyridoxal phosphate may at least in part be responsible. Very large doses of vitamin B6 may be necessary to compensate for this hyd...

  2. Acid Hydrolysis of Bromazepam Catalyzed by Micelles, Reverse Micelles, and Microemulsions


    Ferdousi Begum; Mollah, M. Yousuf A.; M. Muhibur Rahman; Md. Abu Bin Hasan Susan


    Kinetics of the acid hydrolysis of bromazepam (Bz) has been investigated in micelles, reverse micelles, and microemulcions of cetyltrimethylammonium bromide (CTAB) by spectrophotometric method. The rate of the acid hydrolysis of Bz was found to be enhanced both below and above the critical micelle concentration (CMC) of CTAB in aqueous solution. The pseudo-first-order rate constant (k′) shows an initial decrease for both low and high H+ concentrations. With further increase in [CTAB], at low ...

  3. The hydrolysis of cationic polycarboxybetaine esters to zwitterionic polycarboxybetaines with controlled properties. (United States)

    Zhang, Zheng; Cheng, Gang; Carr, Louisa R; Vaisocherová, Hana; Chen, Shengfu; Jiang, Shaoyi


    In this work, we report a new class of materials, cationic polycarboxybetaine esters, which have unique properties when they interact with proteins, DNAs, and bacteria. These cationic polymers can be converted to nontoxic and nonfouling zwitterionic polymers upon their hydrolysis. Due to their unique properties, they are very promising for a wide range of applications, such as highly effective gene delivery carriers and environmentally friendly antimicrobial coatings. Three positively charged polyacrylamides, of which the pedant groups bear carboxybetaine ester groups, were synthesized. These three polymers have different spacer groups between the quaternary ammonium and the ester groups. Their hydrolysis behaviors were studied using proton NMR under different NaOH concentrations. Their interactions with biomolecules and microorganisms before and after hydrolysis were demonstrated by protein adsorption/resistance, DNA condensation/release, and antimicrobial properties. The polymers were grafted onto a gold-coated surface covered with initiators using surface-initiated atom transfer radical polymerization (ATRP). Fibrinogen adsorption was measured by surface plasmon resonance (SPR) sensors. While the polymer-grafted surfaces have high protein adsorption, the surfaces became nonfouling after hydrolysis. Linear polymers were also synthesized and DNA/polymer complexes were evaluated. Agarose gel electrophoresis shows that DNA can be condensed into nanoparticles by the cationic polymers before hydrolysis and released from the DNA/polymer complexes upon the hydrolysis of the cationic polymers into zwitterionic polymers. The complexes formed were characterized by dynamic light scattering measurements. In addition, the interactions of linear polymers with bacteria were also evaluated. The polycarboxybetaine ester with a pentene spacer exhibits evident antimicrobial properties when they are incubated with Gram negative bacteria (Escherichia Coli). The polymer can be

  4. Tailored and integrated production of carboxylated cellulose nanocrystals (CNC) with nanofibrils (CNF) through maleic acid hydrolysis (United States)

    Ruibin Wang; Liheng Chen; J.Y. Zhu; Rendang Yang


    This study demonstrates the feasibility of tailored and integrated production of carboxylated cellulose nanocrystals (CNC) with nanofibrils (CNF) from bleached pulp fibers through hydrolysis using a recyclable dicarboxylic acid. Hydrolysis experiments were conducted using ranges of 15–75 wt% maleic acid concentrations, 60–120°C temperatures, and 5–300 min reaction...

  5. Internal Hydrolysis Indicator for Sample Specific Monitoring of β-Glucuronidase Activity. (United States)

    Taylor, Lacy L; Flint, Noah A; Ma, Vinh; Hill, Brandy M; Clark, Chantry J; Strathmann, Frederick G


    Metabolized forms of benzodiazepines (benzos) can cause issues with mass spectrometry identification. Benzodiazepines undergo a process called glucuronidation during metabolism that attaches a glucuronic acid for increased solubility. Often in clinical testing an enzymatic hydrolysis step is implemented to increase the sensitivity of benzodiazepines by hydrolyzing β-D-glucuronic acid from benzodiazepine-glucuronide conjugates in urine samples using the β-Glucuronidase enzyme. In this study resorufin β-D-glucuronide, a substrate of the β-Glucuronidase enzyme, was added to patient samples to determine if proper hydrolysis had occurred. The presence of resorufin as an Internal Hydrolysis Indicator (IHI) shows the activity and efficiency of the enzyme in each patient sample. Synthetic/patient urine samples were obtained and mixed with hydrolysis buffer containing resorufin β-D-glucuronide. The β-Glucuronidase enzyme was used to hydrolyze the benzodiazepine analytes as well as resorufin β-D-glucuronide. The enzymatic hydrolysis addition increased the positivity rate of benzodiazepines by 42.5%. The β-Glucuronidase substrate resorufin (IHI) displayed variability in area counts between patient samples. Comparative studies with internal standards and resorufin (IHI) showed no correlation between recovery and analyte variability. Hydrolysis reactions greatly improved the sensitivity of benzodiazepines by liquid chromatography time-of-flight mass spectrometry analysis. The large variation in resorufin (IHI) area counts amongst patient samples indicates possible variability in enzymatic hydrolysis activity. The enzymatic hydrolysis step is a part of the extraction procedure and should be controlled for in each patient sample. © The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please email:

  6. Sweetening syrup production by enzymatic hydrolysis of starch variety yam (Dioscorea rotundata)


    Carlos Ramón Vidal Tovar; Gisela Páez


    Sweeteners syrups produced by enzymatic hydrolysis from starch of hawthorn yam (Dioscorea rotundata). The starch was extracted by a scratched, washed, sedimented and drying; the yield was quantified taking into account the amount of initial raw material and was determined the concentration of starch, amylose, amylopectin, crude fiber, ash, protein, fat and humidity in accordance with the requirements of the AOAC standards, and ICONTEC COVENIN. Enzymatic hydrolysis of starch was conducted usin...

  7. Ultrasound assisted acid catalyzed lactose hydrolysis: Understanding into effect of operating parameters and scale up studies. (United States)

    Gajendragadkar, Chinmay N; Gogate, Parag R


    The current work deals with the value addition of lactose by transforming into hydrolyzed lactose syrup containing glucose and galactose in major proportion using the novel approach of ultrasound assisted acid catalyzed lactose hydrolysis. The hydrolysis of lactose was performed in ultrasonic bath (33kHz) at 50% duty cycle at different temperatures as 65°C and 70°C and two different hydrochloric acid (HCl) concentrations as 2.5N and 3N. It was observed that acid concentration, temperature and ultrasonic treatment were the major factors in deciding the time required to achieve ∼90% hydrolysis. The ultrasonic assisted approach resulted in reduction in the reaction time and the extent of intensification was established to be dependent on the temperature, acid concentration and time of ultrasonic exposure. It was observed that the maximum process intensification obtained by introduction of ultrasound in the lactose hydrolysis process performed at 70°C and 3N HCl was reduction in the required time for ∼90% hydrolysis from 4h (without the presence of ultrasound) to 3h. The scale-up study was also performed using an ultrasonic bath with longitudinal horn (36kHz as operating frequency) at 50% duty cycle, optimized temperature of 70°C and acid concentration of 3N. It was observed that the reaction was faster in the presence of ultrasound and stirring by axial impeller at rpm of 225±25. The time required to complete ∼90% of hydrolysis remained almost the same as observed for small scale study on ultrasonic bath (33kHz) at 50% duty cycle. The use of recovered lactose from whey samples instead of pure lactose did not result in any significant changes in the progress of hydrolysis, confirming the efficacy of the selected approach. Overall, the work has presented a novel ultrasound assisted approach for intensified lactose hydrolysis. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. Bile acids stimulate ATP hydrolysis in the purified cholesterol transporter ABCG5/G8. (United States)

    Johnson, Brandy J Harvey; Lee, Jyh-Yeuan; Pickert, Amanda; Urbatsch, Ina L


    ABCG5 and ABCG8 are half-size ABC transporters that function as heterodimers (ABCG5/G8) to reduce sterol absorption in the intestines and increase sterol excretion from the liver. Previous studies demonstrated that bile acids increased ABCG5/G8 specific cholesterol efflux in cell models. In this study we tested the effects of bile acids on ATP hydrolysis in Pichia pastoris purified ABCG5/G8 and found that they stimulated hydrolysis approximately 20-fold in wild-type ABCG5/G8 but not in a hydrolysis-deficient mutant. Nonconjugated cholate supported the highest ATPase activity in ABCG5/G8 (256 +/- 9 nmol min(-1) mg(-1)). ATP hydrolysis was also stimulated by other conjugated bile acids and a mixture of bile acids resembling human bile with activities ranging from 129 +/- 4 to 147 +/- 14 nmol min(-1) mg(-1). The kinetic parameters, inhibitor profiles, and lipid requirements of bile acid stimulated ATP hydrolysis were characterized. Cholate-stimulated ATP hydrolysis was maximal at concentrations of >or=10 mM MgATP and had a relatively high K(M) (MgATP) of approximately 1 mM. Orthovanadate, BeFx, and AlFx effectively inhibited ABCG5/G8 at concentrations of 1 mM. Various lipid mixtures supported bile acid-stimulated ATP hydrolysis, which increased when cholesterol was present. The data demonstrate that bile acids together with lipids and cholesterol increase ATP hydrolysis in purified ABCG5/G8. Bile acids may promote an active conformation of purified ABCG5/G8 either by global stabilization of the transporter or by binding to a specific site on ABCG5/G8.

  9. Effect of high hydrostatic pressure on the enzymatic hydrolysis of bovine serum albumin. (United States)

    De Maria, Serena; Ferrari, Giovanna; Maresca, Paola


    The extent of enzymatic proteolysis mainly depends on accessibility of the peptide bonds, which stabilize the protein structure. The high hydrostatic pressure (HHP) process is able to induce, at certain operating conditions, protein displacement, thus suggesting that this technology can be used to modify protein resistance to the enzymatic attack. This work aims at investigating the mechanism of enzymatic hydrolysis assisted by HHP performed under different processing conditions (pressure level, treatment time). Bovine serum albumin was selected for the experiments, solubilized in sodium phosphate buffer (25 mg mL-1 , pH 7.5) with α-chymotrypsin or trypsin (E/S ratio = 1/10) and HPP treatment (100-500 MPa, 15-25 min). HHP treatment enhanced the extent of the hydrolysis reaction of globular proteins, being more effective than conventional hydrolysis. At HHP treatment conditions maximizing the protein unfolding, the hydrolysis degree of proteins was increased as a consequence of the increased exposure of peptide bonds to the attack of proteolytic enzymes. The maximum hydrolysis degree (10% and 7% respectively for the samples hydrolyzed with α-chymotrypsin and trypsin) was observed for the samples processed at 400 MPa for 25 min. At pressure levels higher than 400 MPa the formation of aggregates was likely to occur; thus the degree of hydrolysis decreased. Protein unfolding represents the key factor controlling the efficiency of HHP-assisted hydrolysis treatments. The peptide produced under high pressure showed lower dimensions and a different structure with respect to those of the hydrolysates obtained when the hydrolysis was carried out at atmospheric pressure, thus opening new frontiers of application in food science and nutrition. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  10. Effect of defatting on acid hydrolysis rate of maize starch with different amylose contents. (United States)

    Wei, Benxi; Hu, Xiuting; Zhang, Bao; Li, Hongyan; Xu, Xueming; Jin, Zhengyu; Tian, Yaoqi


    The effect of defatting on the physiochemical properties and the acid hydrolysis rate of maize starch with different amylose contents was evaluated in this study. The increase in the number of pores and the stripping of starch surface layers were observed after defatting by scanning electron microscopy. X-ray diffraction spectrum showed that the peaks attributing to the amylose-lipid complex disappeared. The relative crystallinity increased by 19% for high-amylose maize starch (HMS) on defatting, while the other tested starches virtually unchanged. Differential scanning calorimetry study indicated an increase in the thermal stability for the defatted starches. Compared with native waxy maize starch, the acid hydrolysis rate of the defatted one increased by 6% after 10 days. For normal maize starch (NMS) and HMS, the higher rate of hydrolysis was observed during the first 5 days. Thereafter, the hydrolysis rate was lower than that of their native counterpart. The increase in susceptibility to acid hydrolysis (in the first 5 days) was mainly attributed to the defective and porous structures formed during defatting process, while the decrease of hydrolysis rate for NMS and HMS samples (after the first 5 days) probably resulted from the increase in the relative crystallinity. Crown Copyright © 2013. Published by Elsevier B.V. All rights reserved.

  11. Efficient Hydrolysis of Rice Straw into Xylose and Glucose by a Two-step Process

    Directory of Open Access Journals (Sweden)

    YAN Lu-lu


    Full Text Available The hydrolysis of rice straw into xylose and glucose in dilute sulfuric acid aqueous solution was studied with a two-step process in batch autoclave reactor. The results showed that compared with the traditional one-step acid hydrolysis, both xylose and glucose could be produced in high yields from rice straw by using the two-step acid hydrolysis process. The effects of reaction temperature, reaction time, the amount of rice straw and acid concentration on the hydrolysis of rice straw were systematically studied, and showed that except initial rice straw loading amount, the other parameters had remarkable influence on the products distribution and yields. In the first-step of the hydrolysis process, a high xylose yield of 162.6 g·kg-1 was obtained at 140℃ after 120 min reaction time. When the solid residues from the first step were subjected to a second-step hydrolysis, a glucose yield as high as 216.5 g·kg-1 could be achieved at 180℃ after 120 min. This work provides a promising strategy for the efficient and value-added utilization of agricultural wastes such as rice straw.

  12. Continuous-flow electro-assisted acid hydrolysis of granular potato starch via inductive methodology. (United States)

    Li, Dandan; Yang, Na; Jin, Yamei; Guo, Lunan; Zhou, Yuyi; Xie, Zhengjun; Jin, Zhengyu; Xu, Xueming


    The induced electric field assisted hydrochloric acid (IEF-HCl) hydrolysis of potato starch was investigated in a fluidic system. The impact of various reaction parameters on the hydrolysis rate, including reactor number (1-4), salt type (KCl, MgCl2, FeCl3), salt concentration (3-12%), temperature (40-55°C), and hydrolysis time (0-60h), were comprehensively assessed. Under optimal conditions, the maximum reducing sugar content in the hydrolysates was 10.59g/L. X-ray diffraction suggested that the crystallinity of IEF-HCl-modified starches increased with the intensification of hydrolysis but was lower than that of native starch. Scanning electron microscopy indicated that the surface and interior regions of starch granules were disrupted by the hydrolysis. The solubility of IEF-HCl-modified starches increased compared to native starch while their swelling power decreased, contributing to a decline in paste viscosity. These results suggest that IEF is a notable potential electrotechnology to conventional hydrolysis under mild conditions without any electrode touching the subject. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. Xylan hydrolysis in Populus trichocarpa × P. deltoides and model substrates during hydrothermal pretreatment. (United States)

    Trajano, Heather L; Pattathil, Sivakumar; Tomkins, Bruce A; Tschaplinski, Timothy J; Hahn, Michael G; Van Berkel, Gary J; Wyman, Charles E


    Previous studies defined easy and difficult to hydrolyze fractions of hemicellulose that may result from bonds among cellulose, hemicellulose, and lignin. To understand how such bonds affect hydrolysis, Populus trichocarpa × Populus deltoides, holocellulose isolated from P. trichocarpa × P. deltoides and birchwood xylan were subjected to hydrothermal flow-through pretreatment. Samples were characterized by glycome profiling, HPLC, and UPLC-MS. Glycome profiling revealed steady fragmentation and removal of glycans from solids during hydrolysis. The extent of polysaccharide fragmentation, hydrolysis rate, and total xylose yield were lowest for P. trichocarpa × P. deltoides and greatest for birchwood xylan. Comparison of results from P. trichocarpa × P. deltoides and holocellulose suggested that lignin-carbohydrate complexes reduce hydrolysis rates and limit release of large xylooligomers. Smaller differences between results with holocellulose and birchwood xylan suggest xylan-cellulose hydrogen bonds limited hydrolysis, but to a lesser extent. These findings imply cell wall structure strongly influences hydrolysis. Copyright © 2014 Elsevier Ltd. All rights reserved.

  14. Kinetic investigation of hydrogen generation from hydrolysis of SnO and Zn solar nanopowders

    Energy Technology Data Exchange (ETDEWEB)

    Chambon, Marc; Abanades, Stephane; Flamant, Gilles [Processes, Materials and Solar Energy Laboratory (CNRS-PROMES), 7 Rue du Four Solaire, 66120 Font-Romeu (France)


    The hydrolysis reaction of the two-step ZnO/Zn and SnO{sub 2}/SnO thermochemical cycles was kinetically investigated for solar hydrogen production. Nanoparticles of Zn and SnO were synthesized by solar thermal reduction of the oxides and neutral gas quenching of the vapors. They were then hydrolyzed to quantify and compare the H{sub 2} yields and the kinetic rate laws in fixed-bed. The hydrolysis of Zn nanoparticles reached only up to 55% of H{sub 2} yield, whereas SnO hydrolysis was almost complete. In contrast, Zn hydrolysis was much faster than SnO hydrolysis, but Zn deactivation occurred suddenly. Models of solid-gas reactions were applied to identify the controlling mechanisms and the associated kinetic parameters. The kinetic models were fitted to both isothermal and non-isothermal (temperature ramp) hydrolysis experimental data. Activation energies and reaction orders were found to be 122 {+-} 13 kJ/mol and 2.0 {+-} 0.3 for SnO, and 87 {+-} 7 kJ/mol and 3.5 {+-} 0.5 for Zn, respectively. Finally, a shrinking core approach was applied to the case of SnO to account for the reaction-controlling mechanisms. (author)

  15. Adsorption of monocomponent enzymes in enzyme mixture analyzed quantitatively during hydrolysis of lignocellulose substrates. (United States)

    Várnai, Anikó; Viikari, Liisa; Marjamaa, Kaisa; Siika-aho, Matti


    The adsorption of purified Trichoderma reesei cellulases (TrCel7A, TrCel6A and TrCel5A) and xylanase TrXyn11 and Aspergillus niger β-glucosidase AnCel3A was studied in enzyme mixture during hydrolysis of two pretreated lignocellulosic materials, steam pretreated and catalytically delignified spruce, along with microcrystalline cellulose (Avicel). The enzyme mixture was compiled to resemble the composition of commercial cellulase preparations. The hydrolysis was carried out at 35 °C to mimic the temperature of the simultaneous saccharification and fermentation (SSF). Enzyme adsorption was followed by analyzing the activity and the protein amount of the individual free enzymes in the hydrolysis supernatant. Most enzymes adsorbed quickly at early stages of the hydrolysis and remained bound throughout the hydrolysis, although the conversion reached was fairly high. Only with the catalytically oxidized spruce samples, the bound enzymes started to be released as the hydrolysis degree reached 80%. The results based on enzyme activities and protein assay were in good accordance. Copyright © 2010 Elsevier Ltd. All rights reserved.

  16. Hydrolysis of Cellulose Using Mono-Component Enzymes Shows Synergy during Hydrolysis of Phosphoric Acid Swollen Cellulose (PASC), but Competition on Avicel

    DEFF Research Database (Denmark)

    Andersen, Natalija; Johansen, Katja S.; Michelsen, Michael Locht


    ). In contrast to previous studies, where P-glucosidase was either not added or added in excess, we here focus on engineering binary, as well as, ternary cellulase mixtures (including a range of different mol% of Cel3A) for maximal total sugar production. Precise hydrolysis pattern based on the concentration...

  17. Enzymatic production of ceramide from sphingomyelin

    DEFF Research Database (Denmark)

    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 potentials in cosmetic and pharmaceutical industries such as in hair and skin care products. Chemical synthesis of ceramide is a costly process, and developments of alternative cost...

  18. Catalytic peptide hydrolysis by mineral surface: Implications for prebiotic chemistry (United States)

    Marshall-Bowman, Karina; Ohara, Shohei; Sverjensky, Dimitri A.; Hazen, Robert M.; Cleaves, H. James


    The abiotic polymerization of amino acids may have been important for the origin of life, as peptides may have been components of the first self-replicating systems. Though amino acid concentrations in the primitive oceans may have been too dilute for significant oligomerization to occur, mineral surface adsorption may have provided a concentration mechanism. As unactivated amino acid polymerization is thermodynamically unfavorable and kinetically slow in aqueous solution, we studied mainly the reverse reaction of polymer degradation to measure the impact of mineral surface catalysis on peptide bonds. Aqueous glycine (G), diglycine (GG), diketopiperazine (DKP), and triglycine (GGG) were reacted with minerals (calcite, hematite, montmorillonite, pyrite, rutile, or amorphous silica) in the presence of 0.05 M, pH 8.1, KHCO 3 buffer and 0.1 M NaCl as background electrolyte in a thermostatted oven at 25, 50 or 70 °C. Below 70 °C, reaction kinetics were too sluggish to detect catalytic activity over amenable laboratory time-scales. Minerals were not found to have measurable effects on the degradation or elongation of G, GG or DKP at 70 °C in solution. At 70 °C pyrite was the most catalytic mineral with detectible effects on the degradation of GGG, although several others also displayed catalytic behavior. GGG degraded ˜1.5-4 times faster in the presence of pyrite than in control reactions, depending on the ratio of solution concentration to mineral surface area. The rate of pyrite catalysis of GGG hydrolysis was found to be saturable, suggesting the presence of discrete catalytic sites on the mineral surface. The mineral-catalyzed degradation of GGG appears to occur via a GGG → DKP + G mechanism, rather than via GGG → GG + G, as in solution-phase reactions. These results are compatible with many previous findings and suggest that minerals may have assisted in peptide synthesis in certain geological settings, specifically by speeding the approach to equilibrium

  19. Effect of Varying Acid Hydrolysis Condition in Gracilaria Sp. Fermentation Using Sasad (United States)

    Mansuit, H.; Samsuri, M. D. C.; Sipaut, C. S.; Yee, C. F.; Yasir, S. M.; Mansa, R.


    Macroalgae or seaweed is being considered as promising feedstock for bioalcohol production due to high polysaccharides content. Polysaccharides can be converted into fermentable sugar through acid hydrolysis pre-treatment. In this study, the potential of using carbohydrate-rich macroalgae, Gracilaria sp. as feedstock for bioalcohol production via various acid hydrolysis conditions prior to the fermentation process was investigated and evaluated. The seaweed used in this research was from the red algae group, using species of Gracilaria sp. which was collected from Sg. Petani Kedah, Malaysia. Pre-treatment of substrate was done using H2SO4 and HCl with molarity ranging from 0.2M to 0.8M. The pretreatment time were varied in the range of 15 to 30 minutes. Fermentation was conducted using Sasad, a local Sabahan fermentation agent as a starter culture. Alcohol extraction was done using a distillation unit. Reducing sugar analysis was done by Benedict test method. Alcohol content analysis was done using specific gravity test. After hydrolysis, it was found out that acid hydrolysis at 0.2M H2SO4 and pre-treated for 20 minutes at 121°C has shown the highest reducing sugar content which has yield (10.06 mg/g) of reducing sugar. It was followed by other samples hydrolysis using 0.4M HCl with 30 minutes pre-treatment and 0.2M H2SO4, 15 minutes pre-treatment with yield of 8.06 mg/g and 5.75 mg/g reducing sugar content respectively. In conclusion, acid hydrolysis of Gracilaria sp. can produce higher reducing sugar yield and thus it can further enhance the bioalcohol production yield. Hence, acid hydrolysis of Gracilaria sp. should be studied more as it is an important step in the bioalcohol production and upscaling process.


    Directory of Open Access Journals (Sweden)

    Y. S. Novikova


    Full Text Available The development of biotechnology fucose and fucooligosaccharides from vegetable raw materials is of particular interest, due to the wide range of biological activity fucose. The fucose is an important carbohydrate component of immunoglobulins and performs important functions in the biological processes of ontogenesis, cell differentiation as well as in the reproductive processes in vertebrates and in the formation of immunity. The fucose and its polymers have prebiotic effects, fucooligosaccharides have also exhibit antioxidant activity. One of the promising methods of obtaining fucose and fucooligosaccharides is an enzymatic hydrolysis of fucoidan vegetable raw materials. The most accessible and inexpensive plant raw materials to extract fucoidan are the brown algae. Literary analysis showed that the algae Fucus vesiculosis are characterized by the highest content of fucoidan, the chemical structure of which is a homopolymer of fucose with a predominance of 1,3 glycoside bonds. It was investigated the hydrolysis of fucoidan algae Fucus vesiculosis by immobilized enzyme preparation α-L-fucosidase for determine the optimal conditions of the enzymatic process. The efficiency of enzymatic degradation of fucoidan depends on several factors, the most important of which are the amount of the enzyme preparation, the pH of the reaction medium and the temperature at which the hydrolysis takes place, as well as its duration. Were determined the optimal process parameters of the enzymatic hydrolysis of fucoidan algal Fucus vesiculosis: the amount of the enzyme preparation of 6 U / g, the temperature 50 ° C, pH 7.0, the hydrolysis time 5 hours. Under these conditions, the degree of hydrolysis of fucoidan was 83-85%, which shows prospects of the enzymatic hydrolysis for obtaining of fucose from vegetable raw materials.

  1. Defining the role of ATP hydrolysis in mitotic segregation of bacterial plasmids.

    Directory of Open Access Journals (Sweden)

    Yoan Ah-Seng

    Full Text Available Hydrolysis of ATP by partition ATPases, although considered a key step in the segregation mechanism that assures stable inheritance of plasmids, is intrinsically very weak. The cognate centromere-binding protein (CBP, together with DNA, stimulates the ATPase to hydrolyse ATP and to undertake the relocation that incites plasmid movement, apparently confirming the need for hydrolysis in partition. However, ATP-binding alone changes ATPase conformation and properties, making it difficult to rigorously distinguish the substrate and cofactor roles of ATP in vivo. We had shown that mutation of arginines R36 and R42 in the F plasmid CBP, SopB, reduces stimulation of SopA-catalyzed ATP hydrolysis without changing SopA-SopB affinity, suggesting the role of hydrolysis could be analyzed using SopA with normal conformational responses to ATP. Here, we report that strongly reducing SopB-mediated stimulation of ATP hydrolysis results in only slight destabilization of mini-F, although the instability, as well as an increase in mini-F clustering, is proportional to the ATPase deficit. Unexpectedly, the reduced stimulation also increased the frequency of SopA relocation over the nucleoid. The increase was due to drastic shortening of the period spent by SopA at nucleoid ends; average speed of migration per se was unchanged. Reduced ATP hydrolysis was also associated with pronounced deviations in positioning of mini-F, though time-averaged positions changed only modestly. Thus, by specifically targeting SopB-stimulated ATP hydrolysis our study reveals that even at levels of ATPase which reduce the efficiency of splitting clusters and the constancy of plasmid positioning, SopB still activates SopA mobility and plasmid positioning, and sustains near wild type levels of plasmid stability.

  2. Intensification of enzymatic hydrolysis of waste newspaper using ultrasound for fermentable sugar production. (United States)

    Subhedar, Preeti B; Babu, Narmadha R; Gogate, Parag R


    An effective conversion of lignocellulose into fermentable sugars is a key step in producing bioethanol in an eco-friendly and cost effective manner. In this study, the effect of ultrasound on enzymatic hydrolysis of newspaper, a potential feedstock for bioethanol production due to its high cellulosic content, was investigated. The effect of substrate loading, enzyme loading, temperature, ultrasonic power and duty cycle on the hydrolysis has been studied. Optimum conditions for conventional enzymatic hydrolysis were substrate loading of 5% (w/v), enzyme loading of 0.14% (w/v), temperature of 323K, and under these conditions and 72h of hydrolysis, reducing sugar yield of 11.569g/L was obtained. In case of ultrasound-assisted enzymatic hydrolysis approach, optimum conditions obtained were substrate loading of 3% (w/v), enzyme loading of 0.8% (w/v), sonication power of 60W, duty cycle of 70%, hydrolysis time of 6.5h and the reducing sugar yield obtained under these conditions was 27.6g/L. Approximately 2.4 times increase in the release of reducing sugar concentration was obtained by the ultrasound-assisted enzymatic hydrolysis approach. Results indicate that there is a synergistic effect obtained from the combination of ultrasound and enzymes which lowers the diffusion-limiting barrier to enzyme/substrate binding and results in an increase in reaction rate. The experimental data were also fitted in a simple three parameter kinetic model. Copyright © 2014 Elsevier B.V. All rights reserved.

  3. Transcriptomic Analysis of THP-1 Macrophages Exposed to Lipoprotein Hydrolysis Products Generated by Lipoprotein Lipase. (United States)

    Thyagarajan, Narmadaa; Marshall, Jenika D; Pickett, Arthur T; Schumacher, Clemens; Yang, Yanbo; Christian, Sherri L; Brown, Robert J


    Macrophage lipoprotein lipase (LPL) induces lipid accumulation and promotes atherosclerosis. However, the effects of lipoprotein hydrolysis products generated by LPL on macrophage-derived foam cell formation are not clearly understood. Thus, we analyzed the transcriptomic response to hydrolysis products via microarray analyses on RNA isolated from human THP-1 macrophages incubated with total lipoprotein hydrolysis products generated by LPL. The expression of 183 transcripts was significantly upregulated and 133 transcripts were significantly downregulated. Bioinformatics analyses revealed that there was a significant over-representation of genes involved in cell cycling, stress response, type I interferon signaling, cellular metal ion homeostasis, sterol metabolism, and nuclease activity. Interestingly, transcripts for 63 small nucleolar RNA were significantly upregulated. We verified the microarray data by quantitative real-time PCR and found that the expression of SNORA56, as well as the expression of genes associated with the cell cycle (PCNA and DKC1 variant 3), stress response (ATF3), type I interferon signaling (IFITM1), and lipid metabolism (CD36 and PLIN2) were significantly affected by LPL hydrolysis products. To determine if the free fatty acid (FFA) component of total lipoprotein hydrolysis products is sufficient to alter the expression of these genes, THP-1 macrophages were also incubated with the total FFA or individual classes of the FFA component. The gene regulation by the FFA component did not mimic that of the hydrolysis products, suggesting that the regulation of gene expression in THP-1 macrophages depends on the specific combination and concentration of lipid species present in the hydrolysis products, and not solely on FFA.

  4. Pseudomonas aeruginosa arylsulfatase: a purified enzyme for the mild hydrolysis of steroid sulfates. (United States)

    Stevenson, Bradley J; Waller, Christopher C; Ma, Paul; Li, Kunkun; Cawley, Adam T; Ollis, David L; McLeod, Malcolm D


    The hydrolysis of sulfate ester conjugates is frequently required prior to analysis for a range of analytical techniques including gas chromatography-mass spectrometry (GC-MS). Sulfate hydrolysis may be achieved with commercial crude arylsulfatase enzyme preparations such as that derived from Helix pomatia but these contain additional enzyme activities such as glucuronidase, oxidase, and reductase that make them unsuitable for many analytical applications. Strong acid can also be used to hydrolyze sulfate esters but this can lead to analyte degradation or increased matrix interference. In this work, the heterologously expressed and purified arylsulfatase from Pseudomonas aeruginosa is shown to promote the mild enzyme-catalyzed hydrolysis of a range of steroid sulfates. The substrate scope of this P. aeruginosa arylsulfatase hydrolysis is compared with commercial crude enzyme preparations such as that derived from H. pomatia. A detailed kinetic comparison is reported for selected examples. Hydrolysis in a urine matrix is demonstrated for dehydroepiandrosterone 3-sulfate and epiandrosterone 3-sulfate. The purified P. aeruginosa arylsulfatase contains only sulfatase activity allowing for the selective hydrolysis of sulfate esters in the presence of glucuronide conjugates as demonstrated in the short three-step chemoenzymatic synthesis of 5α-androstane-3β,17β-diol 17-glucuronide (ADG, 1) from epiandrosterone 3-sulfate. The P. aeruginosa arylsulfatase is readily expressed and purified (0.9 g per L of culture) and thus provides a new and selective method for the hydrolysis of steroid sulfate esters in analytical sample preparation. Copyright © 2015 John Wiley & Sons, Ltd.

  5. [The development of the method for enzymatic hydrolysis for the extraction of toxic substances from the hair samples]. (United States)

    Slustovskaya, Yu V; Krys'ko, M V; Strelova, O Yu

    The objective of the present study was to develop and validate the method for the extraction of toxic substances from the hair samples as exemplified by enzymatic hydrolysis of barbituric acid derivatives. The experiments were carried out with the use of laboratory animals (white female rats and albino guinea pigs) that had been daily given a phenobarbital solution per os during 4 months preceding the study. The hairs obtained from the experimental animals were subjected to acid hydrolysis with a 6 mole hydrochloric acid and enzymatic hydrolysis with the use of chymopsin, trypsin, chymotrypsin, and papain solutions. The analysis of the extracted materials was performed by means of gas chromatography with mass-selective detection. The application of the proposed method for enzymatic hydrolysis produced the better results than acid hydrolysis. This technique was validated. The results of the study made possible the comparative characteristic of the effectiveness of acid and enzymatic hydrolysis.

  6. Surfactant protein B inhibits secretory phospholipase A2 hydrolysis of surfactant phospholipids (United States)

    Grier, Bonnie L.; Waite, B. Moseley; Veldhuizen, Ruud A.; Possmayer, Fred; Yao, Li-Juan; Seeds, Michael C.


    Hydrolysis of surfactant phospholipids (PL) by secretory phospholipases A2 (sPLA2) contributes to surfactant damage in inflammatory airway diseases such as acute lung injury/acute respiratory distress syndrome. We and others have reported that each sPLA2 exhibits specificity in hydrolyzing different PLs in pulmonary surfactant and that the presence of hydrophilic surfactant protein A (SP-A) alters sPLA2-mediated hydrolysis. This report tests the hypothesis that hydrophobic SP-B also inhibits sPLA2-mediated surfactant hydrolysis. Three surfactant preparations were used containing varied amounts of SP-B and radiolabeled tracers of phosphatidylcholine (PC) or phosphatidylglycerol (PG): 1) washed ovine surfactant (OS) (pre- and postorganic extraction) compared with Survanta (protein poor), 2) Survanta supplemented with purified bovine SP-B (1–5%, wt/wt), and 3) a mixture of dipalmitoylphosphatidylcholine (DPPC), 1-palmitoyl-2-oleoyl-phosphatidylcholine (POPC), and 1-palmitoyl-2-oleoyl-phosphatidylglycerol (POPG) (DPPC:POPC:POPG, 40:40:20) prepared as vesicles and monomolecular films in the presence or absence of SP-B. Hydrolysis of PG and PC by Group IB sPLA2 (PLA2G1A) was significantly lower in the extracted OS, which contains SP-B, compared with Survanta (P = 0.005), which is SP-B poor. Hydrolysis of PG and PC in nonextracted OS, which contains all SPs, was lower than both Survanta and extracted OS. When Survanta was supplemented with 1% SP-B, PG and PC hydrolysis by PLA2G1B was significantly lower (P hydrolysis by both PLA2G1B and Group IIA sPLA2 (PLA2G2A). In films, PLA2G1B hydrolyzed surfactant PL monolayers at surface pressures ≤30 mN/m (P hydrolysis can occur. These results suggest the hydrophobic SP, SP-B, protects alveolar surfactant PL from hydrolysis mediated by multiple sPLA2 in both vesicles (alveolar subphase) and monomolecular films (air-liquid interface). PMID:22037357

  7. Monitoring of soluble starch hydrolysis induced by α-amylase from Aspergillus oryzae using ultrasonic spectroscopy (United States)

    Sierra, Carlos; Resa, Pablo; Buckin, Vitaly; Elvira, Luis


    The online monitoring of enzymatic starch hydrolysis is an important issue for several industrial sectors, mainly in the alimentary industry. Ultrasonic non-invasive methods based on the detection of wave velocity and amplitude changes can be used to study this enzymatic reaction. These wave propagating changes are result of physicalchemical modifications produced in the media by the starch hydrolysis. In this work the starch hydrolysis induced by the enzyme α-amylase from Aspergillus oryzae is studied. This biochemical reaction has been monitored using a high-resolution ultrasonic spectroscopy (HR-US) which is non-invasive and nondestructive. The measured time profiles o of ultrasonic velocity are explained in terms of the starch hydrolysis and the subsequent production of oligosaccharides as a consequence of the enzymatic action. The obtained results have been compared to a conventional off-line technique used in biochemistry, the iodine-starch reaction, a spectrophotometric method to quantify the amount of starch remaining in the medium. The combination of these two types of measurement provides more complete information about the biochemical processes occurred during hydrolysis.

  8. Effects of partial hydrolysis and subsequent cross-linking on wheat gluten physicochemical properties and structure. (United States)

    Wang, Kaiqiang; Luo, Shuizhong; Cai, Jing; Sun, Qiaoqiao; Zhao, Yanyan; Zhong, Xiyang; Jiang, Shaotong; Zheng, Zhi


    The rheological behavior and thermal properties of wheat gluten following partial hydrolysis using Alcalase and subsequent microbial transglutaminase (MTGase) cross-linking were investigated. The wheat gluten storage modulus (G') and thermal denaturation temperature (Tg) were significantly increased from 2.26 kPa and 54.43°C to 7.76 kPa and 57.69°C, respectively, by the combined action of partial hydrolysis (DH 0.187%) and cross-linking. The free SH content, surface hydrophobicity, and secondary structure analysis suggested that an appropriate degree of Alcalase-based hydrolysis allowed the compact wheat gluten structure to unfold, increasing the β-sheet content and surface hydrophobicity. This improved its molecular flexibility and exposed additional glutamine sites for MTGase cross-linking. SEM images showed that a compact 3D network formed, while SDS-PAGE profiles revealed that excessive hydrolysis resulted in high-molecular-weight subunits degrading to smaller peptides, unsuitable for cross-linking. It was also demonstrated that the combination of Alcalase-based partial hydrolysis with MTGase cross-linking might be an effective method for modifying wheat gluten rheological behavior and thermal properties. Copyright © 2015 Elsevier Ltd. All rights reserved.

  9. Effects of acid hydrolysis intensity on the properties of starch/xanthan mixtures. (United States)

    Jiang, Min; Hong, Yan; Gu, Zhengbiao; Cheng, Li; Li, Zhaofeng; Li, Caiming


    The effects of acid hydrolysis intensity on the physicochemical properties of starch/xanthan gum (XG) system were studied. Waxy corn starch (WCS) was subjected to different concentrations of hydrochloric acid, and crystallization and relative molecular weight analysis were performed. The results revealed that the starch granules became smaller during acid hydrolysis. X-ray diffraction pattern analysis showed that the crystal structure did not change with acid hydrolysis. Evaluation of the properties and digestibility of different acid-thinned starch/XG systems indicated that the viscosity of acid-thinned starch/XG decreased with increased acid hydrolysis intensity. Rheological property measurements indicated that the compound systems were a pseudo-plastic fluid, which is a typical weak gel structure. Finally, we show that the WCS1.0M/XG has the highest stability of the tested mixtures. We conclude that adjusting the conditions of acid hydrolysis improves the stability and food quality-enhancing properties of starch. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Effect of Enzymatic Hydrolysis on the Antioxidant Properties of Alcoholic Extracts of Oilseed Cakes

    Directory of Open Access Journals (Sweden)

    Jacek Arct


    Full Text Available The aim of the present study is to compare changes in the total phenolic, flavonoid and reducing sugar content and antioxidant activity of alcoholic extracts of Oenothera biennis, Borago officinalis and Nigella sativa oilseed cakes before and after enzymatic hydrolysis. Extraction with ethanol and hydrolysis with different commercially available glycosidases: α-amylase, β-glucosidase, β-glucanase and their combinations in a ratio of 1:1:1 were investigated. Total phenolic, flavonoid and reducing sugar content, iron-chelating activity and antioxidant activity according to DPPH and ABTS tests were measured in non-hydrolysed extracts and compared with the results obtained for the extracts after the application of immobilised enzymes. As a result, the hydrolysed extracts had a higher phenolic and reducing sugar content as well as higher iron-chelating and antioxidant activities. Total phenolic content of Oenothera biennis, Borago officinalis and Nigella sativa oilseed cake extracts after enzymatic hydrolysis was higher in comparison with non-hydrolysed extracts, i.e. 2 times (for the enzyme combination, and 1.5 and 2 times (for β-glucanase (p<0.05, respectively. The best results in increasing the flavonoid and sugar content as well as in iron-chelating activity were obtained after enzymatic hydrolysis of oilseed cake extracts by β-glucanase. Oilseed cake extracts after hydrolysis with an enzyme combination in a ratio of 1:1:1 had the highest increase in antioxidant activity.

  11. Hydrolysis of terbufos using simulated environmental conditions: rates, mechanisms, and product analysis. (United States)

    Hong, F; Win, K Y; Pehkonen, S O


    This study focuses on the hydrolysis of terbufos, an organophosphorus pesticide. Combining GC-MS and wet chemistry methods, di-tert-butyl disulfide and formaldehyde were identified and quantified as major degradation products. Diethyl dithiophosphate was also indirectly identified as a degradation product under alkaline conditions. Hydrolysis rate constants of terbufos under homogeneous conditions were comparable to those of phorate and show relative insensitivity to pH under slightly acidic to neutral pH conditions, as the observed rate constants varied only in the range of (4.5-5.0) x 10(-6) s(-1) between pH 5.7 and 9.4; neutral hydrolysis is thus the most dominant hydrolysis pathway of terbufos in ambient waters. The mechanisms for terbufos hydrolysis and the formation of the major products and their temporal profiles are discussed. To assess the environmental impact of degradation products of this widely used pesticide, Microtox was used to analyze the toxicity of terbufos and two of its degradation products: diethyl dithiophosphate and di-tert-butyl disulfide; the EC(50) of terbufos was found to be >17 microM, whereas the EC(50) of di-tert-butyl disulfide was 1.3 microM.

  12. The characteristic and dispersion stability of nanocellulose produced by mixed acid hydrolysis and ultrasonic assistance. (United States)

    Niu, Fuge; Li, Mengya; Huang, Qi; Zhang, Xiuzhen; Pan, Weichun; Yang, Jiansheng; Li, Jianrong


    Axiolitic shape nanocellulose particles were prepared using a combined mixed acid hydrolysis and ultrasonic treatment. The crystallinity, morphology and stability properties of cellulose were characterized to investigate the mechanism of nanocellulose formation and stability. It was found the hydrodynamic radius decreased from 205nm to 89nm, and the crystallinity index of the nanocellulose increased from 62.90% to 72.31% with an increase in hydrolysis time from 2 to 10h. Sulfate esters and sulfonate group were present in the nanocellulose, and released more COH groups after hydrolysis. The ζ-potential of cellulose decreased from -11.5 to -43.8mV after 10h of hydrolysis. These results illustrated the amorphous characteristic of cellulose was removed after acid hydrolysis and ultrasonic treatment. The higher ζ-potential and relatively small cellulose particles caused a more stable suspension, suggesting that electrostatic interactions played an important role in maintaining the stability and dispersibility of the nanocellulose particles. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. Preparations and mechanism of hydrolysis of ((8)annulene)actinide compounds. [Uranocene

    Energy Technology Data Exchange (ETDEWEB)

    Moore, R.M. Jr.


    The mechanism of hydrolysis for bis(8)annulene actinide and lanthanide complexes has been studied in detail. The uranium complex, uranocene, decomposes with good pseudo-first order kinetics (in uranocene) in 1 M degassed solutions of H/sub 2/O in THF. Decomposition of a series of aryl-substituted uranocenes demonstrates that the hydrolysis rate is dependent on the electronic nature of the substituent (Hammett rho value = 2.1, r/sup 2/ = 0.999), with electron-withdrawing groups increasing the rate. When D/sub 2/O is substituted for H/sub 2/O, kinetic isotope effects of 8 to 14 are found for a variety of substituted uranocenes. These results suggest a pre-equilibrium involving approach of a water molecule to the central metal, followed by rate determining proton transfer to the eight membered ring and rapid decomposition to products. Each of the four protonations of the complex has a significant isotope effect. The product ratio of cyclooctatriene isomers formed in the hydrolysis varies, depending on the central metal of the complex. However, the general mechanism of hydrolysis, established for uranocene, can be extended to the hydrolysis and alcoholysis of all the (8)annulene complexes of the lanthanides and actinides.

  14. Evaluation of hyper thermal acid hydrolysis of Kappaphycus alvarezii for enhanced bioethanol production. (United States)

    Ra, Chae Hun; Nguyen, Trung Hau; Jeong, Gwi-Taek; Kim, Sung-Koo


    Hyper thermal (HT) acid hydrolysis of Kappaphycus alvarezii, a red seaweed, was optimized to 12% (w/v) seaweed slurry content, 180mM H2SO4 at 140°C for 5min. The maximum monosaccharide concentration of 38.3g/L and 66.7% conversion from total fermentable monosaccharides of 57.6g/L with 120gdw/L K. alvarezii slurry were obtained from HT acid hydrolysis and enzymatic saccharification. HT acid hydrolysis at a severity factor of 0.78 efficiently converted the carbohydrates of seaweed to monosaccharides and produced a low concentration of inhibitory compounds. The levels of ethanol production by separate hydrolysis and fermentation with non-adapted and adapted Kluyveromyces marxianus to high concentration of galactose were 6.1g/L with ethanol yield (YEtOH) of 0.19 at 84h and 16.0g/L with YEtOH of 0.42 at 72h, respectively. Development of the HT acid hydrolysis process and adapted yeast could enhance the overall ethanol fermentation yields of K. alvarezii seaweed. Copyright © 2016 Elsevier Ltd. All rights reserved.

  15. Lactose Hydrolysis in Milk and Dairy Whey Using Microbial β-Galactosidases

    Directory of Open Access Journals (Sweden)

    Michele Dutra Rosolen


    Full Text Available This work aimed at evaluating the influence of enzyme concentration, temperature, and reaction time in the lactose hydrolysis process in milk, cheese whey, and whey permeate, using two commercial β-galactosidases of microbial origins. We used Aspergillus oryzae (at temperatures of 10 and 55°C and Kluyveromyces lactis (at temperatures of 10 and 37°C β-galactosidases, both in 3, 6, and 9 U/mL concentrations. In the temperature of 10°C, the K. lactis β-galactosidase enzyme is more efficient in the milk, cheese whey, and whey permeate lactose hydrolysis when compared to A. oryzae. However, in the enzyme reaction time and concentration conditions evaluated, 100% lactose hydrolysis was not reached using the K. lactis β-galactosidase. The total lactose hydrolysis in whey and permeate was obtained with the A. oryzae enzyme, when using its optimum temperature (55°C, at the end of a 12 h reaction, regardless of the enzyme concentration used. For the lactose present in milk, this result occurred in the concentrations of 6 and 9 U/mL, with the same time and temperature conditions. The studied parameters in the lactose enzymatic hydrolysis are critical for enabling the application of β-galactosidases in the food industry.

  16. Fermentable sugar production from paddy straw by two steps chemical pretreatment and hydrolysis process (United States)

    Lee, Vivian J. Q.; Salimi, M. N.; Yusoff, Ahm


    Paddy straw is one of the most abundant lignocellulose wastes and has potential as a feedstock for sugar production. In this study, dilute acid pretreatment and enzymatic hydrolysis are the process that selected for the production of sugar. The paddy straw was pretreated with 1% (v/v) of sulfuric acid at 95 °C for 60 minutes followed by enzymatic hydrolysis. Optimization of enzymatic hydrolysis is desirable to achieve high yield of sugar from solid residues with high cellulose content. The optimization has been carried out by using Central composite design (CCD) to analyze the effect of pH, temperature and enzyme dosing for the enzymatic hydrolysis. For both the process, the concentration of sugar was analyzed by using Dinitrosalicylic acid (DNS) reagent with the aid of the standard glucose curve. The results showed the highest sugar yield from dilute acid pre-treatment was 1.14 g/L. For the optimum condition for enzymatic hydrolysis was at pH 5, 50 °C and 0.10 mL of enzyme and this produced 4.55 g/L of fermentable sugar.

  17. Mechanistic insights into the hydrolysis of 2-chloroethyl ethyl sulfide: the expanded roles of sulfonium salts. (United States)

    Bae, Su Y; Winemiller, Mark D


    The hydrolysis of 2-chloroethyl ethyl sulfide has been examined in an effort to better understand its mechanism under more concentrated conditions. Two salts formed during hydrolysis were synthesized, and an emphasis was placed on determining their effect on the reaction as it proceeded. Unexpected changes in mechanism were seen when excess chloride was added to the reaction. By measuring rates and product distributions as the products were added back into the hydrolysis, a mechanism was developed. The formation of these sulfonium salts represents additional products in the disappearance of 2-chloroethyl ethyl sulfide with k3 in particular causing a deviation away from expected first-order behavior. Sulfonium salts 3 and 4 do not appear to interconvert, and the system as a whole had fewer pathways available than previously proposed. Initial conditions for studying the hydrolysis were very important and could lead to different conclusions depending on the conditions used. This work will aid in better understanding the hydrolysis of the very toxic chemical warfare agent mustard (bis(2-chloroethyl)sulfide) in the environment and during its decontamination.

  18. Hydrolysis of Virgin Coconut Oil Using Immobilized Lipase in a Batch Reactor

    Directory of Open Access Journals (Sweden)

    Lee Suan Chua


    Full Text Available Hydrolysis of virgin coconut oil (VCO had been carried out by using an immobilised lipase from Mucor miehei (Lipozyme in a water-jacketed batch reactor. The kinetic of the hydrolysis was investigated by varying the parameters such as VCO concentration, enzyme loading, water content, and reaction temperature. It was found that VCO exhibited substrate inhibition at the concentration more than 40% (v/v. Lipozyme also achieved the highest production of free fatty acids, 4.56 mM at 1% (w/v of enzyme loading. The optimum water content for VCO hydrolysis was 7% (v/v. A relatively high content of water was required because water was one of the reactants in the hydrolysis. The progress curve and the temperature profile of the enzymatic hydrolysis also showed that Lipozyme could be used for free fatty acid production at the temperature up to 50°C. However, the highest initial reaction rate and the highest yield of free fatty acid production were at 45 and 40°C, respectively. A 100 hours of initial reaction time has to be compensated in order to obtain the highest yield of free fatty acid production at 40°C.

  19. Powerful peracetic acid-ionic liquid pretreatment process for the efficient chemical hydrolysis of lignocellulosic biomass. (United States)

    Uju; Goto, Masahiro; Kamiya, Noriho


    The aim of this work was to design a new method for the efficient saccharification of lignocellulosic biomass (LB) using a combination of peracetic acid (PAA) pretreatment with ionic liquid (IL)-HCl hydrolysis. The pretreatment of LBs with PAA disrupted the lignin fractions, enhanced the dissolution of LB and led to a significant increase in the initial rate of the IL-HCl hydrolysis. The pretreatment of Bagasse with PAA prior to its 1-buthyl-3-methylimidazolium chloride ([Bmim][Cl])-HCl hydrolysis, led to an improvement in the cellulose conversion from 20% to 70% in 1.5h. Interestingly, the 1-buthyl-3-methylpyridium chloride ([Bmpy][Cl])-HCl hydrolysis of Bagasse gave a cellulose conversion greater than 80%, with or without the PAA pretreatment. For LB derived from seaweed waste, the cellulose conversion reached 98% in 1h. The strong hydrolysis power of [Bmpy][Cl] was attributed to its ability to transform cellulose I to II, and lowering the degree of polymerization of cellulose. Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. Kinetic study of the thermal hydrolysis of Agave salmiana for mezcal production. (United States)

    Garcia-Soto, M J; Jimenez-Islas, H; Navarrete-Bolanos, J L; Rico-Martinez, R; Miranda-Lopez, R; Botello-Alvarez, J E


    The kinetics of the thermal hydrolysis of the fructans of Agave salmiana were determined during the cooking step of mezcal production in a pilot autoclave. Thermal hydrolysis was achieved at different temperatures and cooking times, ranging from 96 to 116 °C and from 20 to 80 h. A simple kinetic model of the depolymerization of fructans to monomers and other reducing sugars and of the degradation of reducing sugars to furans [principally 5-(hydroxymethyl)furfural, HMF] was developed. From this model, the rate constants of the reactions were calculated, as well as the pre-exponential factors and activation energies of the Arrhenius equation. The model was found to fit the experimental data well. The tradeoff between a maximum fructan hydrolysis and a critical furan concentration in allowing for the best ethanol yield during fermentation was investigated. The results indicated that the thermal hydrolysis of agave was optimal, from the point of view of ethanol yield in the ensuing fermentation, in the temperature range of 106-116 °C and the cooking range time of 6-14 h. The optimal conditions corresponded to a fructan hydrolysis of 80%, producing syrups with furan and reducing sugar concentrations of 1 ± 0.1 and 110 ± 10 g/L, respectively.

  1. Hydrolysis of macromolecular components of primary and secondary wastewater sludge by thermal hydrolytic pretreatment. (United States)

    Wilson, Christopher A; Novak, John T


    A laboratory simulation of the thermal hydrolytic pretreatment (THP) process was performed on wastewater sludge, as well as key macromolecular components: proteins, lipids, and polysaccharides. Hydrolysis temperatures from 130 to 220 degrees C were investigated. The objectives of this study were to determine how and over which temperature range THP specifically affects sludge components, and whether hydrolysis temperature can be used to minimize the previously reported drawbacks of THP such as high total ammonia nitrogen (TAN) loads and the production of highly-colored recalcitrant organics. In addition, the applicability of THP to primary sludge (PS) was investigated. The breakdown of proteins, lipids, and polysaccharides was determined to be temperature dependent, and both waste activated sludge (WAS) and PS responded similarly to THP apart from intrinsic differences in lipid and protein content. Pure carbohydrate solutions were not largely converted to mono- or dimeric reducing sugar units at temperatures below 220 degrees C, however significant caramelization of starch and production of dextrose and maltose was observed to occur at 220 degrees C. Volatile fatty acid production during thermal hydrolysis was largely attributed to the breakdown of unsaturated lipids, and long-chain fatty acid production was not significant in terms of previous reports of methanogenic inhibition. Ammonia was produced from protein during thermal hydrolysis, however solids loading rather than thermal hydrolysis temperature appeared to be a more meaningful control for ammonia levels in downstream anaerobic digestion.

  2. Comparison of sulfuric and hydrochloric acids as catalysts in hydrolysis of Kappaphycus alvarezii (cottonii). (United States)

    Meinita, Maria Dyah Nur; Hong, Yong-Ki; Jeong, Gwi-Taek


    In this study, hydrolysis of marine algal biomass Kappaphhycus alvarezii using two different acid catalysts was examined with the goal of identifying optimal reaction conditions for the formation of sugars and by-products. K. alvarezii were hydrolyzed by autoclave using sulfuric acid or hydrochloric acid as catalyst with different acid concentrations (0.1-1.0 M), substrate concentrations (1.0-13.5%), hydrolysis time (10-90 min) and hydrolysis temperatures (100-130 (°)C). A difference in galactose, glucose, reducing sugar and total sugar content was observed under the different hydrolysis conditions. Different by-product compounds such as 5-hydroxymethylfurfural and levulinic acid were also observed under the different reaction conditions. The optimal conditions for hydrolysis were achieved at a sulfuric acid concentration, temperature and reaction time of 0.2 M, 130 °C and 15 min, respectively. These results may provide useful information for the development of more efficient systems for biofuel production from marine biomass.

  3. Immobilization of yeast inulinase on chitosan beads for the hydrolysis of inulin in a batch system. (United States)

    Singh, R S; Singh, R P; Kennedy, J F


    An extracellular inulinase was partially purified by ethanol precipitation and gel exclusion chromatography from a cell free extract of Kluyveromyces marxianus. Partially purified inulinase exhibited 420 IU/mg specific activity and it was immobilized on chitosan beads. Activity yield of immobilized inulinase was optimized with glutaraldehyde concentration (1-5%), glutaraldehyde treatment time (30-240min), enzyme coupling-time (2-16h) and enzyme loading (5-30 IU) as functions. Under the optimized conditions maximum yield 65.5% of immobilized inulinase was obtained. Maximum hydrolysis of inulin 84.5% and 78.2% was observed at 125rpm after 4h by immobilized and free enzyme, respectively. A retention-time of 4h and 5h was found optimal for the hydrolysis of inulin under agitation (125rpm) by free and immobilized enzyme, respectively. The recycling of the developed immobilized biocatalyst was carried out after 5h of inulin hydrolysis in a batch system. The developed immobilized biocatalyst was successfully used for the hydrolysis of inulin for 14 batches. This is the first report on the immobilization of yeast inulinase on chitosan beads for the hydrolysis of inulin in a batch system. Copyright © 2016 Elsevier B.V. All rights reserved.

  4. Characteristic Studies of Micron Zinc Particle Hydrolysis in a Fixed Bed Reactor

    Directory of Open Access Journals (Sweden)

    Lv Ming


    Full Text Available Zinc fuel is considered as a kind of promising energy sources for marine propeller. As one of the key steps for zinc marine energy power system, zinc hydrolysis process had been studied experimentally in a fixed bed reactor. In this study, we focus on the characteristics of micron zinc particle hydrolysis. The experimental results suggested that the steam inner diffusion is the controlling step of accumulative zinc particles hydrolysis reaction at a relative lower temperature and a relative higher water partial pressure. In other conditions, the chemical reaction kinetics was the controlling step. And two kinds of chemical reaction kinetics appeared in experiments: the surface reaction and the gas-gas reaction. The latter one occurs usually for larger zinc particles and high reaction temperature. Temperature seems to be one of the most important parameters for the dividing of different reaction mechanisms. Several parameters of the hydrolysis process including heating rate, water partial pressure, the particle size and temperature were also studied in this paper. Results show that the initial reaction temperature of zinc hydrolysis in fixed bed is about 410°C. And the initial reaction temperature increases as the heating rate increases and as the water partial pressure decreases. The total hydrogen yield increases as the heating rate decreases, as the water partial pressure increases, as the zinc particle size decreases, and as the reaction temperature increases. A hydrogen yield of more than 81.5% was obtained in the fixed bed experiments.

  5. Hydrolysis of carotenoid esters from Tagetes erecta by the action of lipases from Yarrowia lipolytica. (United States)

    Abdala, Abraham Figueiras; Gallardo, Alfonso Pérez; Olvera, Lorenzo Guevara; Silva, Eleazar Máximo Escamilla


    The present study was conducted to evaluate the feasibility of enzymatic hydrolysis of carotenoid esters from Tagetes erecta using lipases from the yeast of Yarrowia lipolytica, with the aim of obtaining free lutein. The optimal concentrations of seven nutrients, considering the production of lipases relative to biomass (Yp/x) as the response variable, were determined in flask fermentations. In addition, we studied the effect on hydrolysis of growing Y. lipolytica in the presence of the oleoresin of the marigold flower in flask and stirred tank. Furthermore, hydrolysis of the oleoresin using the lipases from this microorganism was compared with the hydrolysis using lipases from Rhizopus oryzae. Cultured in the presence of marigold oleoresin, Y. lipolytica showed an increase in free carotenoids of 12.41% in flask and 8.8% in stirred tank, representing a fourfold and a threefold increase compared to the initial value in the fermentation, respectively. When lipases from the supernatant from both microorganisms were used for only 14 h hydrolysis experiments, a slight increase was achieved compared to a blank. We concluded that carotenoid esters of the oleoresin could not be completely hydrolyzed in 14 h by these lipases, but that growing Y. lipolytica in the presence of marigold oleoresin gives until fourfold production of free carotenoids in 72 h fermentations.

  6. Mechanisms of the stimulatory effects of rhamnolipid biosurfactant on rice straw hydrolysis

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Qiuzhuo; He, Guofu; Xu, Yatong [Department of Environmental Science, East China Normal University, 3663 North Zhongshan Road, Putuo District, Shanghai 200062 (China); Wang, Juan [Department of Environmental Science and Engineering, Harbin Institute of Technology, Harbin 150090 (China); Cai, Weimin [Department of Environmental Science and Engineering, Harbin Institute of Technology, Harbin 150090 (China); School of Environmental Science and Engineering, Shanghai Jiao Tong University, Shanghai 200240 (China)


    Rhamnolipid biosurfactant, as an addition to rice straw hydrolysis bioprocess, could not only stimulate the hydrolysis rate, but also reduce the requirement for large amount of cellulases and promote its recycling process. In this article, through the observation of the changes of cellulases, microorganism, substrate and their mutual functions, the mechanisms of the stimulatory effect of rhamnolipid on rice straw hydrolysis were investigated. The study found that the addition of rhamnolipid increases the activity of {beta}-glucosidase but stabilizes Cel7A activity. The observed results might be the main mechanisms triggering the stimulatory effect of adding biosurfactants on rice straw hydrolysis. Meanwhile, zeta potential of the substrate increased, which could make the resistance of the cell attached to the substrate weaker. This in turn could facilitate easy adhesion and better retention of the microbial cell in the media. Moreover, we discovered that lignin content played an important role in the stimulatory effect of adding rhamnolipid. The adsorption of rhamnolipid biosurfactant prevented unproductive binding of enzymes to lignin. This could be another important mechanism responsible for the stimulatory effects of adding rhamnolipid on rice straw hydrolysis. (author)

  7. Enhanced dewaterability of sludge during anaerobic digestion with thermal hydrolysis pretreatment: New insights through structure evolution. (United States)

    Zhang, Jingsi; Li, Ning; Dai, Xiaohu; Tao, Wenquan; Jenkinson, Ian R; Li, Zhuo


    Comprehensive insights into the sludge digestate dewaterability were gained through porous network structure of sludge. We measured the evolution of digestate dewaterability, represented by the solid content of centrifugally dewatered cake, in high-solids sequencing batch digesters with and without thermal hydrolysis pretreatment (THP). The results show that the dewaterability of the sludge after digestion was improved by 3.5% (±0.5%) for unpretreated sludge and 5.1% (±0.4%) for thermally hydrolyzed sludge. Compared to the unpretreated sludge digestate, thermal hydrolysis pretreatment eventually resulted in an improvement of dewaterability by 4.6% (±0.5%). Smaller particle size and larger surface area of sludge were induced by thermal hydrolysis and anaerobic digestion treatments. The structure strength and compactness of sludge, represented by elastic modulus and fractal dimension respectively, decreased with increase of digestion time. The porous network structure was broken up by thermal hydrolysis pretreatment and was further weakened during anaerobic digestion, which correspondingly improved the dewaterability of digestates. The logarithm of elastic modulus increased linearly with fractal dimension regardless of the pretreatment. Both fractal dimension and elastic modulus showed linear relationship with dewaterability. The rheological characterization combined with the analysis of fractal dimension of sewage sludge porous network structure was found applicable in quantitative evaluation of sludge dewaterability, which depended positively on both thermal hydrolysis and anaerobic digestion. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Subcritical carbon dioxide-water hydrolysis of sugarcane bagasse pith for reducing sugars production. (United States)

    Liang, Jiezhen; Chen, Xiaopeng; Wang, Linlin; Wei, Xiaojie; Wang, Huasheng; Lu, Songzhou; Li, Yunhua


    The aim of present study was to obtain total reducing sugars (TRS) by hydrolysis in subcritical CO2-water from sugarcane bagasse pith (SCBP), the fibrous residue remaining after papermaking from sugarcane bagasse. The optimum hydrolysis conditions were evaluated by L16(4(5)) orthogonal experiments. The TRS yield achieved 45.8% at the optimal conditions: 200°C, 40min, 500rmin(-1), CO2 initial pressure of 1MPa and liquid-to-solid ratio of 50:1. Fourier transform infrared spectrometry and two-dimensional heteronuclear single quantum coherence nuclear magnetic resonance were used to characterize hydrolysis liquor, treated and untreated SCBP, resulting in the removal of hemicelluloses to mainly produce xylose, glucose and arabinose during hydrolysis. The severity factors had no correlation to TRS yield, indicating that the simple kinetic processes of biomass solubilisation cannot perfectly describe the SCBP hydrolysis. The first-order kinetic model based on consecutive reaction was used to obtain rate constants, activation energies and pre-exponential factors. Copyright © 2016 Elsevier Ltd. All rights reserved.

  9. Hydrolysis of cellulose catalyzed by quaternary ammonium perrhenates in 1-allyl-3-methylimidazolium chloride. (United States)

    Wang, Jingyun; Zhou, Mingdong; Yuan, Yuguo; Zhang, Quan; Fang, Xiangchen; Zang, Shuliang


    Quaternary ammonium perrhenates were applied as catalyst to promote the hydrolysis of cellulose in 1-allyl-3-methylimidazolium chloride ([Amim]Cl). The quaternary ammonium perrhenates displayed good catalytic performance for cellulose hydrolysis. Water was also proven to be effective to promote cellulose hydrolysis. Accordingly, 97% of total reduced sugar (TRS) and 42% of glucose yields could be obtained under the condition of using 5mol% of tetramethyl ammonium perrhenate as catalyst, 70μL of water, ca. 0.6mmol of microcrystalline cellulose (MCC) and 2.0g of [Amim]Cl as solvent under microwave irradiation for 30min at 150°C (optimal conditions). The influence of quaternary ammonium cation on the efficiency of cellulose hydrolysis was examined based on different cation structures of perrhenates. The mechanism on perrhenate catalyzed cellulose hydrolysis is also discussed, whereas hydrogen bonding between ReO4 anion and hydroxyl groups of cellulose is assumed to be the key step for depolymerization of cellulose. Copyright © 2015. Published by Elsevier Ltd.

  10. Hydrolysis behavior of regenerated celluloses with different degree of polymerization under microwave radiation. (United States)

    Ni, Jinping; Teng, Na; Chen, Haizhen; Wang, Jinggang; Zhu, Jin; Na, Haining


    This work studied the hydrolysis behavior of regenerated celluloses (RCs) with different degree of polymerization (DP) by using the catalyst of dilute acid under microwave radiation. Results showed that the DP had a considerable influence on hydrolysis of cellulose. The reactivity of RCs was significantly improved when DP was lower than 51. The highest sugar yield of 59.2% was achieved from RC with lowest DP of 23 at 160 °C for 15 min. But the lowest yield of 32.6% was obtained when RC with highest DP of 132 was used. Recrystallization of cellulose was found to hinder the further hydrolysis particularly with the high DP. The effect of recrystallization can be reduced by the decrease of DP of RCs. This research demonstrates that the DP of RCs plays a crucial role on hydrolysis and it provides a preliminary guide based on DP to find a suitable pretreatment method for cellulose hydrolysis. Copyright © 2015 Elsevier Ltd. All rights reserved.

  11. Accelerated anaerobic hydrolysis rates under a combination of intermittent aeration and anaerobic conditions. (United States)

    Jensen, T R; Lastra Milone, T; Petersen, G; Andersen, H R


    Anaerobic hydrolysis in activated return sludge was investigated in laboratory scale experiments to find if intermittent aeration would accelerate anaerobic hydrolysis rates compared to anaerobic hydrolysis rates under strict anaerobic conditions. The intermittent reactors were set up in a 240 h experiment with intermittent aeration (3 h:3 h) in a period of 24 h followed by a subsequent anaerobic period of 24 h in a cycle of 48 h which was repeated five times during the experiment. The anaerobic reactors were kept under strict anaerobic conditions in the same period (240 h). Two methods for calculating hydrolysis rates based on soluble chemical oxygen demand were compared. Two-way analysis of variance with the Bonferroni post-test was performed in order to register any significant difference between reactors with intermittent aeration and strictly anaerobic conditions respectively. The experiment demonstrated a statistically significant difference in favor of the reactors with intermittent aeration showing a tendency towards accelerated anaerobic hydrolysis rates due to application of intermittent aeration. The conclusion of the work is thus that intermittent aeration applied in the activated return sludge process can improve the treatment capacity further in full scale applications.

  12. Progressive structural changes of Avicel, bleached softwood, and bacterial cellulose during enzymatic hydrolysis (United States)

    Kafle, Kabindra; Shin, Heenae; Lee, Christopher M.; Park, Sunkyu; Kim, Seong H.


    A comprehensive picture of structural changes of cellulosic biomass during enzymatic hydrolysis is essential for a better understanding of enzymatic actions and development of more efficient enzymes. In this study, a suite of analytical techniques including sum frequency generation (SFG) spectroscopy, infrared (IR) spectroscopy, x-ray diffraction (XRD), and x-ray photoelectron spectroscopy (XPS) were employed for lignin-free model biomass samples—Avicel, bleached softwood, and bacterial cellulose—to find correlations between the decrease in hydrolysis rate over time and the structural or chemical changes of biomass during the hydrolysis reaction. The results showed that the decrease in hydrolysis rate over time appears to correlate with the irreversible deposition of non-cellulosic species (either reaction side products or denatured enzymes, or both) on the cellulosic substrate surface. The crystallinity, degree of polymerization, and meso-scale packing of cellulose do not seem to positively correlate with the decrease in hydrolysis rate observed for all three substrates tested in this study. It was also found that the cellulose Iα component of the bacterial cellulose is preferentially hydrolyzed by the enzyme than the cellulose Iβ component. PMID:26463274

  13. Sugars and char formation on subcritical water hydrolysis of sugarcane straw. (United States)

    Lachos-Perez, D; Tompsett, G A; Guerra, P; Timko, M T; Rostagno, M A; Martínez, Julian; Forster-Carneiro, T


    Subcritical water has potential as an environmentally friendly solvent for applications including hydrolysis, liquefaction, extraction, and carbonization. Here, we report hydrolysis of sugarcane straw, an abundant byproduct of sugar production, in a semi-continuous reactor at reaction temperatures ranging from 190 to 260°C and at operating pressures of 9 and 16MPa. The target hydrolysis products were total reducing sugars. The main products of sugarcane straw hydrolysis were glucose, xylose, arabinose, and galactose in addition to 5- hydroxymethylfurfural and furfural as minor byproducts. Fourier transform infrared spectroscopy and thermogravimetric analysis provided additional information on the surface and bulk composition of the residual biomass. Char was present on samples treated at temperatures equal to and greater than 190°C. Samples treated at 260°C contained approximately 20wt% char, yet retained substantial hemicellulose and cellulose content. Hydrolysis temperature of 200°C provided the greatest TRS yield while minimizing char formation. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. Production and characterization of cowpea protein hydrolysate with optimum nitrogen solubility by enzymatic hydrolysis using pepsin. (United States)

    Mune Mune, Martin Alain; Minka, Samuel René


    Cowpea is a source of low-cost and good nutritional quality protein for utilization in food formulations in replacement of animal proteins. Therefore it is necessary that cowpea protein exhibits good functionality, particularly protein solubility which affects the other functional properties. The objective of this study was to produce cowpea protein hydrolysate exhibiting optimum solubility by the adequate combination of hydrolysis parameters, namely time, solid/liquid ratio (SLR) and enzyme/substrate ratio (ESR), and to determine its functional properties and molecular characteristics. A Box-Behnken experimental design was used for the experiments, and a second-order polynomial to model the effects of hydrolysis time, SLR and ESR on the degree of hydrolysis and nitrogen solubility index. The optimum hydrolysis conditions of time 208.61 min, SLR 1/15 (w/w) and ESR 2.25% (w/w) yielded a nitrogen solubility of 75.71%. Protein breakdown and the peptide profile following enzymatic hydrolysis were evaluated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and size exclusion chromatography. Cowpea protein hydrolysate showed higher oil absorption capacity, emulsifying activity and foaming ability compared with the concentrate. The solubility of cowpea protein hydrolysate was adequately optimized by response surface methodology, and the hydrolysate showed adequate functionality for use in food. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  15. Combined subcritical water and enzymatic hydrolysis for reducing sugar production from coconut husk (United States)

    Muharja, Maktum; Junianti, Fitri; Nurtono, Tantular; Widjaja, Arief


    Coconut husk wastes are abundantly available in Indonesia. It has a potential to be used into alternative renewable energy sources such as hydrogen using enzymatic hydrolysis followed by a fermentation process. Unfortunately, enzymatic hydrolysis is hampered by the complex structure of lignocellulose, so the cellulose component is hard to degrade. In this study, Combined Subcritical Water (SCW) and enzymatic hydrolysis are applied to enhance fermentable, thereby reducing production of sugar from coconut husk. There were two steps in this study, the first step was coconut husk pretreated by SCW in batch reactor at 80 bar and 150-200°C for 60 minutes reaction time. Secondly, solid fraction from the results of SCW was hydrolyzed using the mixture of pure cellulose and xylanase enzymes. Analysis was conducted on untreated and SCW-treated by gravimetric assay, liquid fraction after SCW and solid fraction after enzymatic hydrolysis using DNS assay. The maximum yield of reducing sugar (including xylose, arabinose glucose, galactose, mannose) was 1.254 gr per 6 gr raw material, representing 53.95% of total sugar in coconut husk biomass which was obtained at 150°C 80 bar for 60 minutes reaction time of SCW-treated and 6 hour of enzymatic hydrolysis using mixture of pure cellulose and xylanase enzymes (18.6 U /gram of coconut husk).

  16. Hydrolysis of solubilized hemicellulose derived from wet-oxidized wheat straw by a mixture of commercial fungal enzyme preparations

    Energy Technology Data Exchange (ETDEWEB)

    Skammelsen Schmidt, Anette; Thomsen, Alle Belinda; Woidemann, Anders [Risoe National Lab. (Denmark); Tenkanen, Maija [VTT Biotechnology and Food Research (Finland)


    The enzymatic hydrolysis of the solubilized hemicellulose fraction from wet-oxidized wheat straw was investigated for quantification purposes. An optimal hydrolysis depends on factors such as composition of the applied enzyme mixture and the hydrolysis conditions (enzyme loading, hydrolysis time, pH-value, and temperature). A concentrated enzyme mixture was used in this study prepared at VTT Biotechnology and Food Research, Finland, by mixing four commercial enzyme preparations. No distinctive pH-value and temperature optima were identified after a prolonged incubation of 24 hours. By reducing the hydrolysis time to 2 hours a temperature optimum was found at 50 deg. C, where a pH-value higher than 5.2 resulted in reduced activity. An enzyme-substrate-volume-ratio of 0.042, a pH-value of 5.0, and a temperature of 50 deg. C were chosen as the best hydrolysis conditions due to an improved monosaccharide yield. The hydrolysis time was chosen to be 24 hours to ensure equilibrium and total quantification. Even under the best hydrolysis conditions, the overall sugar yield from the enzymatic hydrolysis was only 85% of that of the optimal acid hydrolysis. The glucose yield were approximately the same for the two types of hydrolyses, probably due to the high cellulase activity in the VTT-enzyme mixture. For xylose and arabinose the enzymatic hydrolysis yielded only 80% of that of the acid hydrolysis. As the pentoses existed mainly as complex polymers their degradation required many different enzymes, some of which might be missing from the VTT-enzyme mixture. Furthermore, the removal of side-choins from the xylan backbone during the wet-oxidation pretreatment process might enable the hemicellulosic polymers to interact and precipitate, hence, reducing the enzymatic digestibility of the hemicellulose. (au) 8 tabs., 10 ills., 65 refs.

  17. Enhancing enzymatic hydrolysis of coconut husk through Pseudomonas aeruginosa AP 029/GLVIIA rhamnolipid preparation. (United States)

    de Araújo, Cynthia Kérzia Costa; de Oliveira Campos, Alan; de Araújo Padilha, Carlos Eduardo; de Sousa Júnior, Francisco Canindé; do Nascimento, Ruthinéia Jéssica Alves; de Macedo, Gorete Ribeiro; Dos Santos, Everaldo Silvino


    This work investigated the influence of chemical (Triton X-100) and biological surfactant preparation (rhamnolipids) in coconut husk hydrolysis that was subjected to pretreatment with acid-alkali or alkaline hydrogen peroxide. The natural and pretreated biomass was characterized using the National Renewable Energy Laboratory protocol analysis as well as X-ray diffraction and scanning electron microscopy. The results demonstrated that in terms of the total reducing sugars, there was no significant difference between the hydrolysis using Triton X-100 and rhamnolipids, regardless of the pretreatment. A cellulosic conversion value as high as 33.0% was obtained in experiments with rhamnolipids. The coconut husk was observed to be a potential biomass that could produce second generation ethanol, and the rhamnolipid preparation can be used to support for the enzymatic hydrolysis, enhancing the advantage of cellulose conversion into glucose over chemical surfactants because it is an environmentally friendly approach. Copyright © 2017 Elsevier Ltd. All rights reserved.

  18. Enzymatic Hydrolysis of Yellowfin Tuna (Thunnus albacares By-Products Using Protamex Protease

    Directory of Open Access Journals (Sweden)

    Huong Thi My Nguyen


    Full Text Available Long-term proteolysis of tuna by-products (head, viscera and tail by the wide spectrum protease Protamex has been investigated and compared. After hydrolysis, two fractions (soluble aqueous phase and insoluble sludge were collected. Chemical compositions of each fraction and molecular mass distributions of soluble peptides were determined. Degrees of hydrolysis obtained after 12 h of hydrolysis of head, viscera and tail were 32.3, 16.8 and 22.2 %, respectively. Nitrogen recovery in the soluble fractions was 73.6 % for the head, 82.7 % for the viscera and 85.8 % for the tail. Lipid distribution indicated that the majority of lipids remained in the sludge. Such proteolysis appears useful for the production of very different fractions: one rich in peptides of medium to small molecular mass and poor in lipids, and another one containing the insoluble proteins and the majority of lipids.

  19. Evaluation of Chlorella (Chlorophyta as Source of Fermentable Sugars via Cell Wall Enzymatic Hydrolysis

    Directory of Open Access Journals (Sweden)

    Marcoaurélio Almenara Rodrigues


    The algae were cultivated in inorganic medium, collected at the stationary growth phase and centrifuged. The cell pellet was suspended in citrate buffer, pH 4.8 and subjected to 24 hours hydrolysis at 50°C using a cellulases, xylanases, and amylases blend. The measurement of glucose and reducing sugars concentration in the reaction mixture supernatant, on a dry biomass base, showed hydrolysis yields of 2.9% and 5.03% glucose and 4.8% and 8.6% reducing sugars, for C. homosphaera and C. zofingiensis, respectively. However if cells were washed with chilled ethanol, cold dried, and grounded the biomass hydrolysis yields increased to 23.3% and 18.4% glucose and 24.5% and 19.3% reducing sugars for C. homosphaera and C. zofingiensis, respectively.

  20. Rate of hydrolysis and degradation of the cyanogenic glycoside - dhurrin - in soil

    DEFF Research Database (Denmark)

    Johansen, Henrik; Damgaard, Lars Holm; Olsen, Carl Erik


    in order to assess its fate in soil. The log Kow of dhurrin was 1.18 ± 0.08 (22 C). Hydrolysis was a first-order reaction with respect to dhurrin and hydroxyl ion concentrations. Half lives ranged from 1.2 h (pH 8.6; 25 C) to 530 d (pH 4; 25 C). The activation energy of hydrolysis was 112 + 9 kJ. At pH 5.......8 and room temperature, addition of humic acids (50 g l1) increased the rate of hydrolysis tenfold, while addition of kaolinite or goethite (100–250 g l1) both decreased the rate considerably. No significant sorption to soil components could be observed. The degradation rates of dhurrin in top and subsoils...

  1. Optimal control strategy for fed-batch enzymatic hydrolysis of lignocellulosic biomass based on epidemic modeling. (United States)

    Tai, Chao; Keshwani, Deepak R; Voltan, Diego S; Kuhar, Pankaj S; Engel, Aaron J


    A mathematical optimal control strategy for feeding operation was developed for fed-batch enzymatic hydrolysis of dilute acid pretreated lignocellulosic biomass based on a modified epidemic model. Cellulose conversion was maximized and glucose concentration achieved highest possible value over a fixed hydrolysis time. Boundaries of feeding rate and lignin content were set for feasible controls. Using the optimal control feeding strategy, glucose concentration and accumulated cellulose conversion reached up to 77.31 g/L and 72.08% in 100 h, which are 108.76% and 37.50% higher than in batch hydrolysis with same amount of enzyme consumption. Solids content in feeding source has a significant interference on system mass transfer. Optimal control is a useful tool for guiding operations in fed-batch and continuous processes as it enables process optimization through clear objective functions and feasible controls. © 2015 Wiley Periodicals, Inc.

  2. Hydrolysis of Chlorella by Cellulomonas sp. YJ5 cellulases and its biofunctional properties. (United States)

    Yin, Li-Jung; Jiang, Shann-Tzong; Pon, Shen-Hwei; Lin, Hsin-Hung


    Both 10% and 20% (w/w) Chlorella suspensions were hydrolyzed by 150 to 350 U/mL of cellulases from a 3-d cultivation of Cellulomonas sp. YJ5. Higher chlorophyll, reducing sugars and soluble proteins, and lower residual insoluble solid were observed on both samples after 30-min hydrolysis by various concentrations of cellulases at 50 °C. Decrease in insoluble solid, increases in soluble proteins, peptides and chlorophyll contents, and microscopic observation indicated obvious lysis of cell walls occurred during 60- to 180-min hydrolysis. Significant increases in soluble proteins, peptides, Fe(2+) chelating ability, trolox equivalent antioxidation capacity (TEAC), and reducing power was obtained after 3-h hydrolysis by 150 U/mL of cellulase. These data suggested that cellulolysis technology has high application potential in Chlorella industry. © 2010 Institute of Food Technologists®

  3. Ethanol production by enzymatic hydrolysis: parametric analysis of a base-case process

    Energy Technology Data Exchange (ETDEWEB)

    Isaacs, S.H.


    A base-case flowsheet for an enzymatic hydrolysis process is presented. Included is a parametric sensitivity analysis to identify key research issues and an assessment of this technology. The plant discussed is a large-scale facility, producing 50 million gallons of ethanol per year. The plant design is based on the process originally conceived by the US National Army Command and consists of these process steps: pretreatment; enzyme production; enzyme hydrolysis; fermentation; and distillation. The base-case design parameters are based on recent laboratory data from Lawrence Berkeley Laboratories and the University of California at Berkeley. The selling price of ethanol is used to compare variations in the base-case operating parameters, which include hydrolysis efficiencies, capital costs, enzyme production efficiencies, and enzyme recycle. 28 references, 38 figures, 8 tables.

  4. Kinetics of Enzymatic High-Solid Hydrolysis of Lignocellulosic Biomass Studied by Calorimetry

    DEFF Research Database (Denmark)

    Olsen, Søren Nymand; Rasmussen, Erik Lumby; McFarland, K.C.


    . In the current work, we have investigated the application of isothermal calorimetry to study the kinetics of enzymatic hydrolysis of two substrates (pretreated corn stover and Avicel) at high-solid contents (up to 29% w/w). It was found that the calorimetric heat flow provided a true measure of the hydrolysis...... rate with a detection limit of about 500 pmol glucose s−1. Hence, calorimetry is shown to be a highly sensitive real-time method, applicable for high solids, and independent on the complexity of the substrate. Dose–response experiments with a typical cellulase cocktail enabled a multidimensional...... (excess of attack points) at later stages (>10% conversion). This kinetic profile is interpreted as an increase in polymer end concentration (substrate for CBH) as the hydrolysis progresses, probably due to EG activity in the enzyme cocktail. Finally, irreversible enzyme inactivation did not appear...

  5. Processing of micro-nano bacterial cellulose with hydrolysis method as a reinforcing bioplastic (United States)

    Maryam, Maryam; Dedy, Rahmad; Yunizurwan, Yunizurwan


    Nanotechnology is the ability to create and manipulate atoms and molecules on the smallest of scales. Their size allows them to exhibit novel and significantly improved physical, chemical, biological properties, phenomena, and processes because of their size. The purpose of this research is obtaining micro-nano bacterial cellulose as reinforcing bioplastics. Bacterial cellulose (BC) was made from coconut water for two weeks. BC was dried and grinded. Bacterial cellulose was given purification process with NaOH 5% for 6 hours. Making the micro-nano bacterial cellulose with hydrolysis method. Hydrolysis process with hydrochloric acid (HCl) at the conditions 3,5M, 55°C, 6 hours. Drying process used spray dryer. The hydrolysis process was obtained bacterial cellulose with ±7 μm. The addition 2% micro-nano bacterial cellulose as reinforcing in bioplastics composite can improve the physical characteristics.

  6. Lipase pre-hydrolysis enhance anaerobic biodigestion of soap stock from an oil refining industry. (United States)

    Cherif, Slim; Aloui, Fathi; Carrière, Frédéric; Sayadi, Sami


    A novel alcalophilic Staphylococcus haemolyticus strain with the lipolytic activity was used to perform enzymatic hydrolysis pretreatment of soap stock: a lipid rich solid waste from an oil refining industry. The culture liquid of the selected bacteria and an enzymatic preparation obtained by precipitation with ammonium sulphate from a filtrate of the same culture liquid were used for enzymatic pretreatment. The hydrolysis was carried with different incubation concentrations (10, 20 and 30%) of soap stock and the pretreatment efficiency was verified by running comparative biodegradability tests (crude and treated lipid waste). All pretreated assays showed higher reaction rate compared to crude lipid waste, which was confirmed by the increased levels of biogas production. The pretreatment of solutions containing 10% emulsified soap stock was optimized for 24 h hydrolysis time, enabling high-biogaz formation (800 ml). The use of enzymatic pre-treatment seemed to be a very promising alternative for treating soap stock having high fat contents.

  7. The hydrolysis of geminal ethers: a kinetic appraisal of orthoesters and ketals

    Directory of Open Access Journals (Sweden)

    Sonia L. Repetto


    Full Text Available A novel approach to protecting jet fuel against the effects of water contamination is predicated upon the coupling of the rapid hydrolysis reactions of lipophilic cyclic geminal ethers, with the concomitant production of a hydrophilic acyclic hydroxyester with de-icing properties (Fuel Dehydrating Icing Inhibitors - FDII. To this end, a kinetic appraisal of the hydrolysis reactions of representative geminal ethers was undertaken using a convenient surrogate for the fuel–water interface (D2O/CD3CN 1:4. We present here a library of acyclic and five/six-membered cyclic geminal ethers arranged according to their hydroxonium catalytic coefficients for hydrolysis, providing for the first time a framework for the development of FDII. A combination of 1H NMR, labelling and computational studies was used to assess the effects that may govern the observed relative rates of hydrolyses.

  8. Understanding oscillatory phenomena in molecular hydrogen generation via sodium borohydride hydrolysis. (United States)

    Budroni, M A; Biosa, E; Garroni, S; Mulas, G R C; Marchettini, N; Culeddu, N; Rustici, M


    The hydrolysis of borohydride salts represents one of the most promising processes for the generation of high purity molecular hydrogen under mild conditions. In this work we show that the sodium borohydride hydrolysis exhibits a fingerprinting periodic oscillatory transient in the hydrogen flow over a wide range of experimental conditions. We disproved the possibility that flow oscillations are driven by supersaturation phenomena of gaseous bubbles in the reactive mixture or by a nonlinear thermal feedback according to a thermokinetic model. Our experimental results indicate that the NaBH4 hydrolysis is a spontaneous inorganic oscillator, in which the hydrogen flow oscillations are coupled to an "oscillophor" in the reactive solution. The discovery of this original oscillator paves the way for a new class of chemical oscillators, with fundamental implications not only for testing the general theory on oscillations, but also with a view to chemical control of borohydride systems used as a source of hydrogen based green fuel.

  9. Hydrolysis of monetite/chitosan composites in α-MEM and SBF solutions. (United States)

    Touny, Ahmed H; Dawkins, Herbert; Zhou, Huan; Bhaduri, Sarit B


    There are two objectives of this work. The first objective is to study the hydrolysis behavior of monetite cements formed in the presence and absence of the chitosan in cell culture media (α-MEM) and simulated body fluid (SBF) solutions at 37°C. During hydrolysis, monetite transformed to carbonated apatite. Therefore, the second objective is to examine how addition of chitosan affects on the formation of carbonated apatite phases. The changes in the phase structure of monetite after hydrolysis reactions were characterized using XRD, FTIR and SEM. Pure monetite and monetite/chitosan composite were soaked in α-MEM and SBF solution for 4 and 7 days. In α-MEM solution, the monetite particles started to transform into carbonated apatite with a slow rate. However, in SBF, the rate of monetite transformation to carbonated apatite was more rapid. The presence of the chitosan had no significant effect on the precipitation of carbonated apatite on the monetite particles.

  10. Microtubule dynamic instability: A new model with coupled GTP hydrolysis and multistep catastrophe (United States)

    Bowne-Anderson, Hugo; Zanic, Marija; Kauer, Monika; Howard, Jonathon


    A key question in understanding microtubule dynamics is how GTP hydrolysis leads to catastrophe, the switch from slow growth to rapid shrinkage. We first provide a review of the experimental and modeling literature, and then present a new model of microtubule dynamics. We demonstrate that vectorial, random, and coupled hydrolysis mechanisms are not consistent with the dependence of catastrophe on tubulin concentration and show that, although single-protofilament models can explain many features of dynamics, they do not describe catastrophe as a multistep process. Finally, we present a new combined (coupled plus random hydrolysis) multiple-protofilament model that is a simple, analytically solvable generalization of a single-protofilament model. This model accounts for the observed lifetimes of growing microtubules, the delay to catastrophe following dilution and describes catastrophe as a multistep process. PMID:23532586

  11. Influence of thermal hydrolysis pretreatment on organic transformation characteristics of high solid anaerobic digestion. (United States)

    Han, Yun; Zhuo, Yang; Peng, Dangcong; Yao, Qian; Li, Huijuan; Qu, Qiliang


    The study evaluated the influence of thermal hydrolysis pretreatment (THP) on anaerobic digestion (AD) ability of high solid sludge. The transformation characteristics of organics during the THP+AD process of dewatering sludge from wastewater treatment plant was investigated using a lab-scale THP reactor and four anaerobic digesters. The reduction efficiency of volatile suspended solids using THP+AD exceeded 49%. The acceleration of biogas production during AD was due to the enhancement of protein hydrolysis and acidogenesis by THP. THP had only minimal influence on the improvement of carbohydrate acidogenesis. The hydrolysis of poly phosphates was likely the main reaction of phosphorus transformation. Biochemical generation of sulfide and ammonia nitrogen occurred during the acidogenesis. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. Enhanced enzymatic cellulose hydrolysis by subcritical carbon dioxide pretreatment of sugarcane bagasse. (United States)

    Zhang, Hongdan; Wu, Shubin


    Most biomass pretreatment processes for sugar production are run at low-solid concentration (carbon dioxide (CO2) could provide a more sustainable pretreatment medium while using relative high-solid contents (15 wt.%). The effects of subcritical CO2 pretreatment of sugarcane bagasse to the solid and glucan recoveries at different pretreatment conditions were investigated. Subsequently, enzymatic hydrolysis at different hydrolysis time was applied to obtain maximal glucose yield, which can be used for ethanol fermentation. The maximum glucose yield in enzyme hydrolyzate reached 38.5 g based on 100g raw material after 72 h of enzymatic hydrolysis, representing 93.0% glucose in sugarcane bagasse. The enhanced digestibilities of subcritical CO2 pretreated sugarcane bagasse were due to the removal of hemicellulose, which were confirmed by XRD, FTIR, SEM, and TGA analyses. Copyright © 2014 Elsevier Ltd. All rights reserved.

  13. Role of supramolecular cellulose structures in enzymatic hydrolysis of plant cell walls

    DEFF Research Database (Denmark)

    Thygesen, Lisbeth Garbrecht; Hidayat, Budi Juliman; Johansen, Katja Salomon


    The study of biomass deconstruction by enzymatic hydrolysis has hitherto not focussed on the importance of supramolecular structures of cellulose. In lignocellulose fibres, regions with a different organisation of the microfibrils are present. These regions are called dislocations or slip planes...... and they are known to be more susceptible to various forms of degradation such as acid hydrolysis. Traditionally the cellulose within these regions has been assumed to be amorphous, but in this study it is shown by use of polarized light microscopy that dislocations are birefringent. This indicates that they have...... mixture on hydrothermally treated wheat straw, it was found that the fibres were cut into segments corresponding to the sections between the dislocations initially present, as has previously been observed for acid hydrolysis of softwood pulps. The results indicate that dislocations are important during...

  14. Simultaneous hydrolysis and co-fermentation of whey lactose with wheat for ethanol production. (United States)

    Jin, Yiqiong; Parashar, Archana; Mason, Beth; Bressler, David C


    Whey permeate was used as a co-substrate to replace part of the wheat for ethanol production by Saccharomyces cerevisiae. The simultaneous saccharification and fermentation was achieved with β-galactosidase added at the onset of the fermentation to promote whey lactose hydrolysis. Aspergillus oryzae and Kluyveromyces lactis β-galactosidases were two enzymes selected and used in the co-fermentation of wheat and whey permeate for the comparison of their effectiveness on lactose hydrolysis. The possibility of co-fermentations in both STARGEN and jet cooking systems was investigated in 5L bioreactors. Ethanol yields from the co-fermentations of wheat and whey permeate were evaluated. It was found that A. oryzae β-galactosidase was more efficient for lactose hydrolysis during the co-fermentation and that whey permeate supplementation can contribute to ethanol yield in co-fermentations with wheat. Copyright © 2016 Elsevier Ltd. All rights reserved.

  15. A novel approach to monitor the hydrolysis of barley (Hordeum vulgare L) malt: a chemometrics approach. (United States)

    Cozzolino, D; Degner, S; Eglinton, J


    Malting barley is a process that has been profusely studied and is known to be influenced by several physical and biochemical properties of the grain. In particular, the amount of material that can be extracted from the malt (malt extract) is an important measure of brewing performance and end quality. The objectives of this study were (a) to compare the time course of hydrolysis of different malting barley (Hordeum vulgare L) varieties and (b) to evaluate the usefulness of mid-infrared (MIR) spectroscopy as high-throughput method to monitor malt hydrolysis. Differences in the pattern of hydrolysis were observed between the malt samples analyzed where samples from the same variety that have similar hot water extract (HWE) values tend to have the same pattern of hydrolysis. Principal component score plots based on the MIR spectra showed similar results. Partial least-squares discriminate analysis (PLS-DA) was used to classify malt samples according to their corresponding variety and time course of hydrolysis. The coefficient of determination (R(2)) and the standard error of cross validation (SECV) obtained for the prediction of variety and time course of hydrolysis were 0.67 (1.01) and 0.38 (19.90), respectively. These differences might be the result of the different composition in sugars between the barley varieties analyzed after malting, measured as wort density and not observed when only the HWE value at the end point is reported. This method offers the possibility to measure several parameters in malt simultaneously, reducing the time of analysis as well as requiring minimal sample preparation.

  16. Enzymatic Hydrolysis of Pretreated Fibre Pressed Oil Palm Frond by using Sacchariseb C6 (United States)

    Hashim, F. S.; Yussof, H. W.; Zahari, M. A. K. M.; Rahman, R. A.; Illias, R. M.


    Enzymatic hydrolysis becomes a prominent technology for conversion of cellulosic biomass to its glucose monomers that requires an action of cellulolytic enzymes in a sequential and synergistic manner. In this study, the effect of agitation speed, glucan loading, enzyme loading, temperature and reaction time on the production of glucose from fibre pressed oil palm frond (FPOPF) during enzymatic hydrolysis was screened by a half factorial design 25-1 using Response Surface Methodology (RSM). The FPOPF sample was first delignified by alkaline pretreatment at 4.42 (w/v) sodium hydroxide for an hour prior to enzymatic hydrolysis using commercial cellulase enzyme, Sacchariseb C6. The effect of enzymatic hydrolysis on the structural of FPOPF has been evaluated by Scanning Electron Microscopy (SEM) analysis. Characterization of raw FPOPF comprised of 4.5 extractives, 40.7 glucan, 26.1 xylan, 26.2 lignin and 1.8 ash, whereas for pretreated FPOPF gave 0.3 extractives, 61.4 glucan, 20.4 xylan, 13.3 lignin and 1.3 ash. From this study, it was found that the best enzymatic hydrolysis condition yielded 33.01 ± 0.73 g/L of glucose when performed at 200 rpm of agitation speed, 60 FPU/mL of enzyme loading, 4 (w/w) of glucan loading, temperature at 55 □ and 72 hours of reaction time. The model obtained was significant with p-value determination (R2) from ANOVA study was 0.9959. Overall, it can be concluded that addition of Sacchariseb C6 during enzymatic hydrolysis from pretreated FPOPF produce high amount of glucose that enhances it potential for industrial application. This glucose can be further used to produce high-value products.

  17. Barley grain constituents, starch composition, and structure affect starch in vitro enzymatic hydrolysis. (United States)

    Asare, Eric K; Jaiswal, Sarita; Maley, Jason; Båga, Monica; Sammynaiken, Ramaswami; Rossnagel, Brian G; Chibbar, Ravindra N


    The relationship between starch physical properties and enzymatic hydrolysis was determined using ten different hulless barley genotypes with variable carbohydrate composition. The ten barley genotypes included one normal starch (CDC McGwire), three increased amylose starches (SH99250, SH99073, and SB94893), and six waxy starches (CDC Alamo, CDC Fibar, CDC Candle, Waxy Betzes, CDC Rattan, and SB94912). Total starch concentration positively influenced thousand grain weight (TGW) (r(2) = 0.70, p starch concentration (r(2) = -0.80, p hydrolysis of pure starch (r(2) = -0.67, p starch concentration (r(2) = 0.46, p starch (RS) in meal and pure starch samples. The rate of starch hydrolysis was high in pure starch samples as compared to meal samples. Enzymatic hydrolysis rate both in meal and pure starch samples followed the order waxy > normal > increased amylose. Rapidly digestible starch (RDS) increased with a decrease in amylose concentration. Atomic force microscopy (AFM) analysis revealed a higher polydispersity index of amylose in CDC McGwire and increased amylose genotypes which could contribute to their reduced enzymatic hydrolysis, compared to waxy starch genotypes. Increased β-glucan and dietary fiber concentration also reduced the enzymatic hydrolysis of meal samples. An average linkage cluster analysis dendrogram revealed that variation in amylose concentration significantly (p starch concentration in meal and pure starch samples. RS is also associated with B-type granules (5-15 μm) and the amylopectin F-III (19-36 DP) fraction. In conclusion, the results suggest that barley genotype SH99250 with less decrease in grain weight in comparison to that of other increased amylose genotypes (SH99073 and SH94893) could be a promising genotype to develop cultivars with increased amylose grain starch without compromising grain weight and yield.

  18. Development and industrial application of catalyzer for low-temperature hydrogenation hydrolysis of Claus tail gas

    Directory of Open Access Journals (Sweden)

    Honggang Chang


    Full Text Available With the implementation of more strict national environmental protection laws, energy conservation, emission reduction and clean production will present higher requirements for sulfur recovery tail gas processing techniques and catalyzers. As for Claus tail gas, conventional hydrogenation catalyzers are gradually being replaced by low-temperature hydrogenation catalyzers. This paper concentrates on the development of technologies for low-temperature hydrogenation hydrolysis catalyzers, preparation of such catalyzers and their industrial application. In view of the specific features of SO2 hydrogenation and organic sulfur hydrolysis during low-temperature hydrogenation, a new technical process involving joint application of hydrogenation catalyzers and hydrolysis catalyzers was proposed. In addition, low-temperature hydrogenation catalyzers and low-temperature hydrolysis catalyzers suitable for low-temperature conditions were developed. Joint application of these two kinds of catalyzers may reduce the inlet temperatures in the conventional hydrogenation reactors from 280 °C to 220 °C, at the same time, hydrogenation conversion rates of SO2 can be enhanced to over 99%. To further accelerate the hydrolysis rate of organic sulfur, the catalyzers for hydrolysis of low-temperature organic sulfur were developed. In lab tests, the volume ratio of the total sulfur content in tail gas can be as low as 131 × 10−6 when these two kinds of catalyzers were used in a proportion of 5:5 in volumes. Industrial application of these catalyzers was implemented in 17 sulfur recovery tail gas processing facilities of 15 companies. As a result, Sinopec Jinling Petrochemical Company had outstanding application performances with a tail gas discharging rate lower than 77.9 mg/m3 and a total sulfur recovery of 99.97%.

  19. Catalytic hydrolysis of COS over CeO2 (110) surface: A density functional theory study (United States)

    Song, Xin; Ning, Ping; Wang, Chi; Li, Kai; Tang, Lihong; Sun, Xin


    Density functional theory (DFT) calculations were performed to investigate the reaction pathways for catalytic hydrolysis of COS over CeO2 (110) surface using Dmol3 model. The thermodynamic stability analysis for the suggested routes of COS hydrolysis to CO2 and H2S was evaluated. The absolute values of adsorption energy of H2O-CeO2 are higher than that of COS-CeO2. Meanwhile, the adsorption energy and geometries show that H2O is easier adsorbed on the surface of CeO2 (110) than COS. H2O plays a role as a bridge in the process of joint adsorption. H2O forms more Cesbnd Osbnd H groups on the CeO2 (110) surface. CeO2 decreases the maximum energy barrier by 76.15 kcal/mol. The migration of H from H2O to COS is the key for the hydrolysis reaction. Csbnd O channel is easier to occur than Csbnd S channel. Experimental result shows that adding of CeO2 can increase COS removal rate and prolong the 100% COS removal rate from 180 min to 210 min. The difference between Fe2O3 and CeO2 for the hydrolysis of COS is characterized in the atomic charge transfer and the formation of Hsbnd O bond and Hsbnd S bond. The transfer effect of H in H2O to S in COS over CeO2 decreases the energy barriers of hydrolysis reaction, and enhances the reaction activity of COS hydrolysis.

  20. ATP Binding and Hydrolysis Properties of ABCB10 and Their Regulation by Glutathione (United States)

    Qiu, Wei; Liesa, Marc; Carpenter, Elizabeth P.; Shirihai, Orian S.


    ABCB10 (ATP binding cassette sub-family B10) is a mitochondrial inner-membrane ABC transporter. ABCB10 has been shown to protect the heart from the impact of ROS during ischemia-reperfusion and to allow for proper hemoglobin synthesis during erythroid development. ABC transporters are proteins that increase ATP binding and hydrolysis activity in the presence of the transported substrate. However, molecular entities transported by ABCB10 and its regulatory mechanisms are currently unknown. Here we characterized ATP binding and hydrolysis properties of ABCB10 by using the 8-azido-ATP photolabeling technique. This technique can identify potential ABCB10 regulators, transported substrates and amino-acidic residues required for ATP binding and hydrolysis. We confirmed that Gly497 and Lys498 in the Walker A motif, Glu624 in the Walker B motif and Gly602 in the C-Loop motif of ABCB10 are required for proper ATP binding and hydrolysis activity, as their mutation changed ABCB10 8-Azido-ATP photo-labeling. In addition, we show that the potential ABCB10 transported entity and heme precursor delta-aminolevulinic acid (dALA) does not alter 8-azido-ATP photo-labeling. In contrast, oxidized glutathione (GSSG) stimulates ATP hydrolysis without affecting ATP binding, whereas reduced glutathione (GSH) inhibits ATP binding and hydrolysis. Indeed, we detectABCB10 glutathionylation in Cys547 and show that it is one of the exposed cysteine residues within ABCB10 structure. In all, we characterize essential residues for ABCB10 ATPase activity and we provide evidence that supports the exclusion of dALA as a potential substrate directly transported by ABCB10. Last, we show the first molecular mechanism by which mitochondrial oxidative status, through GSH/GSSG, can regulate ABCB10. PMID:26053025

  1. Potential of hydrolysis of particulate COD in extended anaerobic conditions to enhance biological phosphorous removal. (United States)

    Jabari, P; Yuan, Q; Oleszkiewicz, J A


    The effect of anaerobic hydrolysis of particulate COD (pCOD) on biological phosphorous removal in extended anaerobic condition was investigated through (i) sequencing batch reactors (SBR)s with anaerobic hydraulic retention time (HRT) of 0.8, 2, and 4 h; (ii) batch tests using biomass from a full scale biological nutrient removal (BNR) plant; and (iii) activated sludge modeling (BioWin 4.1 simulation). The results from long-term SBRs operation showed that phosphorus removal was correlated to the ratio of filtered COD (FCOD) to total phosphorus (TP) in the influent. Under conditions with low FCOD/TP ratio (average of 20) in the influent, extending anaerobic HRT to 4 h in the presence of pCOD did not significantly improve overall phosphorous removal. During the period with high FCOD/TP ratio (average of 37) in the influent, all SBRs removed phosphorous completely, and the long anaerobic HRT did not have negative effect on overall phosphorous removal. The batch tests also showed that pCOD at different concentration during 4 h test did not affect the rate of anaerobic phosphorus release. The rate of anaerobic hydrolysis of pCOD was significantly low and extending the anaerobic HRT was ineffective. The simulation (BioWin 4.1) of SBRs with low influent FCOD/TP ratio showed that the default kinetics of anaerobic hydrolysis in ASM2d overestimated phosphorous removal in the SBRs (high anaerobic hydrolysis of pCOD). The default anaerobic hydrolysis rate in BioWin 4.1 (ten times lower) could produce similar phosphorous removal to that in the experiment. Results showed that the current kinetics of anaerobic hydrolysis in ASM2d could lead to considerable error in predicting phosphorus removal in processes with extended anaerobic HRT. Biotechnol. Bioeng. 2016;113: 2377-2385. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  2. Hydrogen generation from aqueous acid-catalyzed hydrolysis of sodium borohydride

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Hyun Jae [Clean Fuels and Catalysis Program, The EMS Energy Institute, and Department of Energy and Mineral Engineering, The Pennsylvania State University, University Park, PA 16802 (United States); Shin, Kyoung-Jin; Kim, Hyun-Jong; Han, M.K. [Surface technology team, Korea Institute of Industrial Technology (KITECH), Incheon 406-840 (Korea, Republic of); Kim, Hansung; Shul, Yong-Gun [Department of Chemical and Biomolecular Engineering, Yonsei University, Seoul 120-749 (Korea, Republic of); Jung, Kyeong Taek [Cheil Industries Co. LTD, 437-711 (Korea, Republic of)


    In this study, the hydrogen feed from both Ru-catalyzed and organic acid-catalyzed hydrolysis of NaBH{sub 4} was studied in terms of hydrogen generation rate and integrated PEMFC performance. Hydrogen feed generated from the conventional Ru-catalyzed hydrolysis of NaBH{sub 4} caused a drastic loss of PEMFC performance. It was found that the presence of sodium ion in hydrogen feed was a main factor that increased the interfacial resistance of fuel cell and, consequently, reduced the performance. Acid-catalyzed hydrolysis with powder form of NaBH{sub 4} was adopted in order to minimize the detrimental effect of sodium ion. The hydrogen feed from acid-catalyzed hydrolysis was quite dry so that even water vapor, the carrier of sodium ion, was not detected after condensation of hydrogen feed. It was confirmed by the several experiments that the hydrogen release rate can be controlled by varying the injection rate and concentration of aqueous acid. Various organic acids were employed in the production of hydrogen and found that acidity, acid type and chemical structure are also important factors on hydrolysis of NaBH{sub 4}. The performance from the integrated acid-catalyzed hydrogen generation system with PEMFC was quite stable and no significant loss was observed contrary to that from the integrated Ru-catalyzed hydrogen generation system-PEMFC test. This result also clarified that the detrimental effect of sodium ion could be removed by minimizing the water vapor in this manner. Based on the experiment of acid-catalyzed hydrolysis, a small-scale hydrogen-generating device was designed and fabricated, from which hydrogen release was controlled by the acid concentration and injection rate of aqueous acid solution. (author)

  3. Conformational Footprint in Hydrolysis-Induced Nanofibrillation and Crystallization of Poly(lactic acid). (United States)

    Xu, Huan; Yang, Xi; Xie, Lan; Hakkarainen, Minna


    The origin of hydrolysis-induced nanofibrillation and crystallization, at the molecular level, was revealed by mapping the conformational ordering during long-term hydrolytic degradation of initially amorphous poly(lactic acid) (PLA), a representative model for degradable aliphatic polyesters generally displaying strong interplay between crystallization and hydrolytic erosion. The conformational regularization of chain segments was essentially the main driving force for the morphological evolution of PLA during hydrolytic degradation. For hydrolysis at 37 °C, no significant structural variations were observed due to the immobilization of "frozen" PLA chains. In contrast, conformational ordering in PLA was immediately triggered during hydrolysis at 60 °C and was responsible for the transition from random coils to disordered trans and, further, to quasi-crystalline nanospheres. On the surfaces, the head-by-head absorption and joining of neighboring nanospheres led to nanofibrillar assemblies following a "gluttonous snake"-like manner. The length and density of nanofibers formed were in close relation to the hydrolytic evolution, both of which showed a direct rise in the initial 60 days and then a gradual decline. In the interior, presumably the high surface energy of the nanospheres allowed for the preferential anchoring and packing of conformationally ordered chains into lamellae. In accordance with the well-established hypothesis, the amorphous regions were attacked prior to the erosion of crystalline entities, causing a rapid increase of crystallinity during the initial 30 days, followed by a gradual fall until 90 days. In addition to adequate illustration of hydrolysis-induced variations of crystallinity, our proposed model elucidates the formation of spherulitic nuclei featuring an extremely wide distribution of diameters ranging from several nanometers to over 5 μm, as well as the inferior resistance to hydrolysis observed for the primary nuclei. Our work

  4. Important limitations in the modeling of activated sludge : biased calibration of the hydrolysis process

    DEFF Research Database (Denmark)

    Insel, G.; Gul, Ö.K.; Orhon, D.


    ), provides the majority of the required experimental database. However, currently used procedures still involve a number of basic and practical problems. Model evaluation of the OUR data may generate a distorted image of the processes involved. Hydrolysis is the most important, yet the most vulnerable...... process as far as the experimental assessment of accurate kinetic parameters is concerned. This study intends to provide an overview of major experimental limitations in the modeling of activated sludge, with emphasis on the appropriate experimental design for the assessment of the hydrolysis rate....

  5. Assessing Reliability of Cellulose Hydrolysis Models to Support Biofuel Process Design – Identifiability and Uncertainty Analysis

    DEFF Research Database (Denmark)

    Sin, Gürkan; Meyer, Anne S.; Gernaey, Krist


    The reliability of cellulose hydrolysis models is studied using the NREL model. An identifiability analysis revealed that only 6 out of 26 parameters are identifiable from the available data (typical hydrolysis experiments). Attempting to identify a higher number of parameters (as done in the ori...... to analyze the uncertainty of model predictions. This allows judging the fitness of the model to the purpose under uncertainty. Hence we recommend uncertainty analysis as a proactive solution when faced with model uncertainty, which is the case for biofuel process development research....

  6. Partial-hydrothermal hydrolysis is an effective way to recover bioactives from turmeric wastes

    Directory of Open Access Journals (Sweden)

    Ádina Lima de SANTANA


    Full Text Available Abstract Partial-hydrothermal hydrolysis using pressurized hot water was applied in waste turmeric, derived from extraction processes that employed supercritical and pressurized fluids, as a sustainable alternative for the recovery of bioactive constituents from this material. The reaction products consisted of a liquid extract of fermentable sugars and a solid fraction, which is a mixed biopolymer with relevant composition in terms of starch and antioxidant constituents. The effects of reaction conditions on resulting products were investigated. Economic evaluation was also performed showing that application of partial-hydrothermal hydrolysis from a residual feedstock is feasible in terms of low spend of energy and materials.



    Hamzeh Hoseinpour; Keikhosro Karimi; Hamid Zilouei; Mohammad J. Taherzadeh


    Mixtures of starch and lignocelluloses are available in many industrial, agricultural, and municipal wastes and residuals. In this work, dilute sulfuric acid was used for simultaneous pretreatment of lignocellulose and hydrolysis of starch, to obtain a maximum amount of fermentable sugar after enzymatic hydrolysis with cellulase and β-glucosidase. The acid treatment was carried out at 70-150°C with 0-1% (v/v) acid concentration and 5-15% (w/v) solids concentration for 0-40 minutes. Under the ...

  8. Enzymatic hydrolysis and fermentation of dilute acid pretreated cornstalk to biohydrogen (United States)

    Pan, C. M.; Fan, Y. T.; Hou, H. W.


    The coupling method of acid pretreatment and enzymatic hydrolysis of cornstalk for hydrogen production was investigated in this study. Experimental results showed that temperature, pH and enzyme loading all had an individual significant influence on soluble sugar yield and Ps. The optimum condition for soluble sugar was close to that for Ps. The maximum hydrogen yield from cornstalk by anaerobic mixed microflora was 209.8 ml/g-TVS on the optimum enzymatic hydrolysis condition which was 52 °C of temperature, pH4.8 and 9.4 IU/g of enzyme loading.

  9. Microwave Assisted Hydrolysis of Holocellulose Catalyzed with Sulfonated Char Derived from Lignin-Rich Residue

    Directory of Open Access Journals (Sweden)

    Kui Wang


    Full Text Available A microwave assisted green process has been developed for production of sugars through liquefying holocellulose catalyzed with sulfonated char derived from the lignin-rich residue produced during pretreatment of lignocellulose. Various reaction parameters including the hydrolysis temperature, hydrolysis time, catalyst content, and the ratio of water to feedstock were evaluated. The maximum sugars yield of 82.6% (based on the dry mass of holocellulose was obtained under the optimum reaction conditions. The sulfonated char showed superior catalytic performance to that of dilute sulfuric acid in converting holocellulose into sugars under microwave irradiation.

  10. Method for Improving Separation of Carbohydrates from Wood Pulping Liquors and Wood or Biomass Hydrolysis Liquors

    Energy Technology Data Exchange (ETDEWEB)

    Compere, A L; Marcoccia, B S [Domtar Paper Company, LLC; Elliott, J [General Atomics


    Work with industrial partners to perform the studies needed to commercialize U.S. patent 7,699,958 for separation of carbohydrates from wood pulping liquors and wood or biomass hydrolysis liquors. These include: 1) selection of the best pulp mill liquor withdrawal sites, 2) additional purification or enzyme hydrolysis required to obtain acceptable sugar feedstocks, 3) and work with partners to optimize the stream and purification methods to provide acceptable feedstocks for algal fuels and industrial chemicals production, and 4) preparation of samples large enough for testing by downstream partners.

  11. Aquivion Perfluorosulfonic Superacid as an Efficient Pickering Interfacial Catalyst for the Hydrolysis of Triglycerides. (United States)

    Shi, Hui; Fan, Zhaoyu; Hong, Bing; Pera-Titus, Marc


    Rational design of the surface properties of heterogeneous catalysts can boost the interfacial activity in biphasic reactions through the generation of Pickering emulsions. This concept, termed Pickering interfacial catalysis (PIC), has shown promising credentials in acid-catalyzed transesterification, ester hydrolysis, acetalization, etherification, and alkylation reactions. PIC has now been applied to the efficient, solvent-free hydrolysis of the triglyceride glyceryl trilaurate to lauric acid, catalyzed by Aquivion perfluorosulfonic superacid at mild conditions (100 °C and ambient pressure). © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Hydrolysis mechanism of (N, N) chelated cytotoxic Pt/Pd(II)-dichloro complexes: A theoretical approach (United States)

    Mukherjee, Subhajit; Reddy B., Venkata P.; Mitra, Ishani; Linert, Wolfgang; Moi, Sankar Ch


    Two cytotoxic complexes cis-[Pt(MAMP)Cl2], 1 and cis-[Pd(MAMP)Cl2], 2 (where, MAMP = 2-[(Methylamino)methyl]pyridine) have been considered to explore their hydrolysis mechanism through Density Functional Theory. The stationary states on potential energy surfaces are optimized and characterized. Activation parameters and rate constants for hydrolysis of the complexes have been calculated in CPCM model and compared with that of renowned anticancer drugs. Second step is the rate-determining step with greater activation energy for both the complexes. Time Dependent Density Functional Theory is performed in order to understand the nature of electronic transition in the complexes.

  13. The role of acetyl xylan esterase in the solubilization of xylan and enzymatic hydrolysis of wheat straw and giant reed

    Directory of Open Access Journals (Sweden)

    Zhang Junhua


    Full Text Available Abstract Background Due to the complexity of lignocellulosic materials, a complete enzymatic hydrolysis into fermentable sugars requires a variety of cellulolytic and xylanolytic enzymes. Addition of xylanases has been shown to significantly improve the performance of cellulases and to increase cellulose hydrolysis by solubilizing xylans in lignocellulosic materials. The goal of this work was to investigate the effect of acetyl xylan esterase (AXE originating from Trichoderma reesei on xylan solubilization and enzymatic hydrolysis of cellulose. Results The solubilization of xylan in pretreated wheat straw and giant reed (Arundo donax by xylanolytic enzymes and the impact of the sequential or simultaneous solubilization of xylan on the hydrolysis of cellulose by purified enzymes were investigated. The results showed that the removal of acetyl groups in xylan by AXE increased the accessibility of xylan to xylanase and improved the hydrolysis of xylan in pretreated wheat straw and giant reed. Solubilization of xylan led to an increased accessibility of cellulose to cellulases and thereby increased the hydrolysis extent of cellulose. A clear synergistic effect between cellulases and xylanolytic enzymes was observed. The highest hydrolysis yield of cellulose was obtained with a simultaneous use of cellulases, xylanase and AXE, indicating the presence of acetylated xylan within the cellulose matrix. Acetylated xylobiose and acetylated xylotriose were produced from xylan without AXE, as confirmed by atmospheric pressure matrix-assisted laser desorption/ionization ion trap mass spectrometry. Conclusions The results in this paper demonstrate that supplementation of xylanase with AXE enhances the solubilization of xylan to some extent and, consequently, increases the subsequent hydrolysis of cellulose. The highest hydrolysis yield was, however, obtained by simultaneous hydrolysis of xylan and cellulose, indicating a layered structure of cellulose and

  14. Lipoprotein lipase-catalyzed hydrolysis of tri[14C]oleoylglycerol in a phospholipid interface : A monolayer study

    NARCIS (Netherlands)

    Demel, R.A.; Shirai, K.; Jackson, R.L.


    The lipoprotein lipase-catalyzed hydrolysis of triacylglycerol was determined in a lipid monolayer containing egg phosphatidylcholine and tri[14C]oleoylglycerol. In the presence of purified bovine milk lipoprotein lipase and fatty acid-free albumin, the rate of hydrolysis of tri[14C]oleoylglycerol,

  15. Prediction of hydrolysis pathways and kinetics for antibiotics under environmental pH conditions: a quantum chemical study on cephradine. (United States)

    Zhang, Haiqin; Xie, Hongbin; Chen, Jingwen; Zhang, Shushen


    Understanding hydrolysis pathways and kinetics of many antibiotics that have multiple hydrolyzable functional groups is important for their fate assessment. However, experimental determination of hydrolysis encounters difficulties due to time and cost restraint. We employed the density functional theory and transition state theory to predict the hydrolysis pathways and kinetics of cephradine, a model of cephalosporin with two hydrolyzable groups, two ionization states, two isomers and two nucleophilic attack directions. Results showed that the hydrolysis of cephradine at pH = 8.0 proceeds via opening of the β-lactam ring followed by intramolecular amidation. The predicted rate constants at different pH conditions are of the same order of magnitude as the experimental values, and the predicted products are confirmed by experiment. This study identified a catalytic role of the carboxyl group in the hydrolysis, and implies that the carboxyl group also plays a catalytic role in the hydrolysis of other cephalosporin and penicillin antibiotics. This is a first attempt to quantum chemically predict hydrolysis of an antibiotic with complex pathways, and indicates that to predict hydrolysis products under the environmental pH conditions, the variation of the rate constants for different pathways with pH should be evaluated.

  16. Temperature dependence of microbial degradation of organic matter in marine sediments: polysaccharide hydrolysis, oxygen consumption, and sulfate reduction

    DEFF Research Database (Denmark)

    Arnosti, C.; Jørgensen, BB; Sagemann, J.


    were incubated in a temperature gradient block spanning a temperature range of ca 45 degrees C. The initial step of organic carbon remineralization, macromolecule hydrolysis, was measured via the enzymatic hydrolysis of fluorescently labeled polysaccharides. The terminal steps of organic carbon...

  17. Introducing enzyme selectivity as a quantitative parameter to describe the effects of substrate concentration on protein hydrolysis

    NARCIS (Netherlands)

    Butré, C.I.


    To understand the differences in peptide composition that result from variations in the conditions of enzymatic hydrolysis of proteins (e.g. substrate concentration) the mechanism of hydrolysis needs to be understood in detail. Therefore, methods and tools were developed to characterize and quantify

  18. Quantitative predictions of bioconversion of aspen by dilute acid and SPORL pretreatments using a unified combined hydrolysis factor (CHF) (United States)

    W. Zhu; Carl J. Houtman; J.Y. Zhu; Roland Gleisner; K.F. Chen


    A combined hydrolysis factor (CHF) was developed to predict xylan hydrolysis during pretreatments of native aspen (Populus tremuloides) wood chips. A natural extension of previously developed kinetic models allowed us to account for the effect of catalysts by dilute acid and two sulfite pretreatments at different pH values....

  19. Efficiencies of acid catalysts in the hydrolysis of lignocellulosic biomass over a range of combined severity factors (United States)

    Jae-Won Lee; Thomas W. Jeffries


    Dicarboxylic organic acids have properties that differ from those of sulfuric acid during hydrolysis of lignocellulose. To investigate the effects of different acid catalysts on the hydrolysis and degradation of biomass compounds over a range of thermochemical pretreatments, maleic, oxalic and sulfuric acids were each used at the same combined severity factor (CSF)...

  20. Hydrolysis of ATP at only one GyrB subunit is sufficient to promote supercoiling by DNA gyrase

    DEFF Research Database (Denmark)

    Kampranis, S C; Maxwell, A


    Mutation of Glu42 to Ala in the B subunit of DNA gyrase abolishes ATP hydrolysis but not nucleotide binding. Gyrase complexes that contain one wild-type and one Ala42 mutant B protein were formed, and the ability of such complexes to hydrolyze ATP was investigated. We found that ATP hydrolysis...

  1. Where does hydrolysis of nandrolone decanoate occur in the human body after release from an oil depot? (United States)

    Kalicharan, R W; Bout, M R; Oussoren, C; Vromans, H


    Long-term therapy of nandrolone (N) is recommended to increase mineral density and muscle strength. Using a parenteral sustained release drug formulation with nandrolone decanoate (ND), therapeutic N levels can be achieved and maintained. Until now, it is unknown if hydrolysis of ND into N occurs in tissue at the injection site or after systemic absorption. Therefore, hydrolysis studies were conducted to investigate the location and rate of ND hydrolysis after its release from the oil depot. ND hydrolysis was studied in porcine tissues, to mimic the human muscular and subcutaneous tissues. Additionally, the ND hydrolysis was studied in human whole blood, plasma and serum at a concentration range of 23.3-233.3μM. ND hydrolysis only occurred in human whole blood. The hydrolysis did not start immediately, but after a lag time. The mean lag time for all studied concentrations was 34.9±2.5min. Because of a slow penetration into tissue, hydrolysis of ND is found to be very low in surrounding tissue. Therefore the local generation of the active compound is clinically irrelevant. It is argued that after injection of the oil depot, ND molecules will be transported via the lymphatic system towards lymph nodes. From here, it will enter the central circulation and within half an hour it will hydrolyse to the active N compound. Copyright © 2016 Elsevier B.V. All rights reserved.

  2. Influence of the degree of hydrolysis of poly(styrene-alt-maleic anhydride) on miscibility with poly(vinyl acetate)

    NARCIS (Netherlands)

    Bosma, M.; Vorenkamp, E.J.; Brinke, G. ten; Challa, G.


    The influence of the hydrolysis of anhydride groups in poly(styrene-alt-maleic anhydride) (PSMA) on its miscibility with poly(vinyl acetate) (PVAc) is investigated. The cloudpoint curves of these blends are determined as a function of the degree of hydrolysis. The miscibility is shown to improve

  3. Exploration of process parameters for continuous hydrolysis of canola oil, camelina oil and algal oil

    KAUST Repository

    Wang, Weicheng


    Thermal hydrolysis of triglycerides to form free fatty acid (FFA) is a well-established industry practice. Recently, this process has been employed as a first step in the production of biofuels from lipids. To that end, batch and continuous hydrolysis of various feedstocks has been examined at the laboratory scale. Canola, the primary feedstock in this paper, camelina and algal oils were converted to high quality FFA. For the different reaction temperatures, the continuous hydrolysis system was found to provide better yields than the laboratory batch system. In addition, CFD simulation with ANSYS-CFX was used to model the performance and reactant/product separation in the continuous, counter-flow reactor. The effects of reaction temperature, water-to-oil ratio (ratio of water and oil volumetric inflow rate), and preheating of the reactants were examined experimentally. Optimization of these parameters has resulted in an improved, continuous process with high mass yields (89-93%, for reactor temperature of 260°C and water-to-oil ratio of 4:1) and energy efficiency (76%, for reactor temperature of 250°C and water-to-oil ratio of 2:1). Based on the product quality and energy efficiency considerations, the reactor temperature of 260°C and water-to-oil ratio of 4:1 have provided the optimal condition for the lab scale continuous hydrolysis reaction. © 2012 Elsevier B.V.

  4. Some observations on the stoichiometry of feldspar hydrolysis in granitic soil (United States)

    James L. Clayton


    Weathering rates of orthoclase and plagioclase were computed from mass balances of Na, K, and Ca in three forested watersheds in the Idaho batholith. On the basis of stand conditions, two watersheds were assumed to have no net gains or losses of cations in biomass, and increases in biomass were measured in the third watershed. Balanced feldspar hydrolysis reactions...

  5. Volcano Plot for Bimetallic Catalysts in Hydrogen Generation by Hydrolysis of Sodium Borohydride (United States)

    Koska, Anais; Toshikj, Nikola; Hoett, Sandra; Bernaud, Laurent; Demirci, Umit B.


    In the field of "hydrogen energy", sodium borohydride (NaBH[subscript 4]) is a potential hydrogen carrier able to release H[subscript 2] by hydrolysis in the presence of a metal catalyst. Our laboratory experiment focuses on this. It is intended for thirdyear undergraduate students in order to have hands-on laboratory experience through…

  6. Modeling enzymatic hydrolysis of lignocellulosic substrates using fluorescent confocal microscopy II: pretreated biomass. (United States)

    Luterbacher, Jeremy S; Moran-Mirabal, Jose M; Burkholder, Eric W; Walker, Larry P


    In this study, we extend imaging and modeling work that was done in Part I of this report for a pure cellulose substrate (filter paper) to more industrially relevant substrates (untreated and pretreated hardwood and switchgrass). Using confocal fluorescence microscopy, we are able to track both the structure of the biomass particle via its autofluorescence, and bound enzyme from a commercial cellulase cocktail supplemented with a small fraction of fluorescently labeled Trichoderma reseii Cel7A. Imaging was performed throughout hydrolysis at temperatures relevant to industrial processing (50°C). Enzyme bound predominantly to areas with low autofluorescence, where structure loss and lignin removal had occurred during pretreatment; this confirms the importance of these processes for successful hydrolysis. The overall shape of both untreated and pretreated hardwood and switchgrass particles showed little change during enzymatic hydrolysis beyond a drop in autofluorescence intensity. The permanence of shape along with a relatively constant bound enzyme signal throughout hydrolysis was similar to observations previously made for filter paper, and was consistent with a modeling geometry of a hollowing out cylinder with widening pores represented as infinite slits. Modeling estimates of available surface areas for pretreated biomass were consistent with previously reported experimental results. © 2014 Wiley Periodicals, Inc.

  7. Constant enthalpy change value during pyrophosphate hydrolysis within the physiological limits of NaCl. (United States)

    Wakai, Satoshi; Kidokoro, Shun-ichi; Masaki, Kazuo; Nakasone, Kaoru; Sambongi, Yoshihiro


    A decrease in water activity was thought to result in smaller enthalpy change values during PPi hydrolysis, indicating the importance of solvation for the reaction. However, the physiological significance of this phenomenon is unknown. Here, we combined biochemistry and calorimetry to solve this problem using NaCl, a physiologically occurring water activity-reducing reagent. The pyrophosphatase activities of extremely halophilic Haloarcula japonica, which can grow at ∼4 M NaCl, and non-halophilic Escherichia coli and Saccharomyces cerevisiae were maximal at 2.0 and 0.1 M NaCl, respectively. Thus, halophilic and non-halophilic pyrophosphatases exhibit distinct maximal activities at different NaCl concentration ranges. Upon calorimetry, the same exothermic enthalpy change of -35 kJ/mol was obtained for the halophile and non-halophiles at 1.5-4.0 and 0.1-2.0 M NaCl, respectively. These results show that solvation changes caused by up to 4.0 M NaCl (water activity of ∼0.84) do not affect the enthalpy change in PPi hydrolysis. It has been postulated that PPi is an ATP analog, having a so-called high energy phosphate bond, and that the hydrolysis of both compounds is enthalpically driven. Therefore, our results indicate that the hydrolysis of high energy phosphate compounds, which are responsible for biological energy conversion, is enthalpically driven within the physiological limits of NaCl.

  8. Preparation and characterization of dialdehyde starch by one-step acid hydrolysis and oxidation. (United States)

    Zuo, Yingfeng; Liu, Wenjie; Xiao, Junhua; Zhao, Xing; Zhu, Ying; Wu, Yiqiang


    Dialdehyde starch was prepared by one-step synthesis of acid hydrolysis and oxidation, using corn starch as the raw material, sodium periodate (NaIO4) as the oxidant, and hydrochloric acid (HCl) as the acid solution. The prepared dialdehyde starch was characterized by Fourier transform infrared spectroscopy (FT-IR), X-ray photoelectron spectroscopy (XPS), and gel permeation chromatography (GPC). The results confirmed that oxidation occurred between the starch and NaIO4. The acid hydrolysis reaction reduced the molecular weight of starch and effectively improved the aldehyde group contents (92.7%). Scanning electron microscope (SEM) analysis indicated that the average particle size decreased after acid hydrolysis and oxidation reaction. X-ray diffraction (XRD) and thermal gravimetric analyzer (TGA) analysis demonstrated that the crystallinity of the obtained dialdehyde starch showed a downward trend and a decelerated thermal decomposition rate. The starch after acid hydrolysis and oxidation exhibited lower hot paste viscosity and higher reactivity. Copyright © 2017. Published by Elsevier B.V.

  9. Cassava Pulp Hydrolysis under Microwave Irradiation with Oxalic Acid Catalyst for Ethanol Production

    Directory of Open Access Journals (Sweden)

    Euis Hermiati


    Full Text Available Microwave irradiation is an alternative method of starch hydrolysis that offers a rapid process. The aim of this research was to improve microwave-assisted hydrolysis of cassava pulp by using oxalic acid as a catalyst. Suspension of cassava pulp in 0.5% oxalic acid (1 g/20 mL was subjected to microwave irradiation at 140-230 °C for 5 minutes, with 4 minutes of pre-heating. One gram of fractured activated carbon made of coconut shell was added into a number of suspensions that were subjected to the same conditions of microwave irradiation. The soluble fraction of the hydrolysates was analyzed for its total soluble solids, malto-oligomer distribution, glucose content, pH value, and formation of brown compounds. The effects of the combined severity parameter at a substrate concentration of 5-12.5% on the glucose yield were also evaluated. The highest glucose yield (78% of dry matter was obtained after hydrolysis at 180 °C without activated carbon addition. Heating above 180 °C reduced the glucose yield and increased the pH and the formation of brown compounds. The use of activated carbon in microwave-assisted acid hydrolysis of cassava pulp reduced the glucose yield, but suppressed the formation of brown compounds. The highest glucose yield (70-80% of dry matter was attained at a severity parameter of 1.3-1.5.

  10. Investigating the Hydrolysis of Starch Using "a"-Amylase Contained in Dishwashing Detergent and Human Saliva (United States)

    Munegumi, Toratane; Inutsuka, Masato; Hayafuji, Yukitaka


    Although saliva has commonly been used to teach about digestion by organisms, the phenomenon of digestion is actually caused by enzymes as catalytic substances. This activity explores the hydrolysis of starch by "a"-amylase in cleaning materials as well as a comparison with the similar reaction using human saliva. The fact that the…

  11. Amylolytic hydrolysis of native starch granules affected by granule surface area. (United States)

    Kim, J C; Kong, B W; Kim, M J; Lee, S H


    Initial stage of hydrolysis of native starch granules with various amylolytic enzymes, alpha-amylase from Bacillus subtilis, glucoamylase I (GA-I) and II (GA-II) from Aspergillus niger, and beta-amylase from sweet potato showed that the reaction was apparently affected by a specific surface area of the starch granules. The ratios of the reciprocal of initial velocity of each amylolytic hydrolysis for native potato and maize starch to that for rice with the amylolytic enzymes were nearly equivalent to the ratio of surface area per mass of the 2 starch granules to that of rice, that is, 6.94 and 2.25, respectively. Thus, the reciprocal of initial velocity of each enzymatic hydrolysis as expressed in a Lineweaver-Burk plot was a linear function of the reciprocal of surface area for each starch granule. As a result, it is concluded that amylolytic hydrolysis of native starch granules is governed by the specific surface area, not by the mass concentration, of each granule.

  12. Starch hydrolysis in autogenerative high pressure digestion: Gelatinisation and saccharification as rate limiting steps

    NARCIS (Netherlands)

    Lindeboom, R.E.F.; Ding, L.; Weijma, J.; Plugge, C.M.; Lier, van J.B.


    Autogenerative high pressure digestion (AHPD) provides an integrated biogas upgrading technology, capable of producing biogas with a CH4 content exceeding 95% at pressures up to 90 bar. Hydrolysis is generally regarded as the rate-limiting step in the anaerobic digestion of complex organic matter,

  13. Determination of the extent of acid hydrolysis of N-methylpyrrolidone. (United States)

    Trusell, F


    The extent of the hydrolysis of N-methylpyrrolidone in dilute hydrochloric acid can readily be determined by titrating an aliquot of the reaction mixture in absolute ethanol with tetrabutylammonium hydroxide. A separate equivalence point is observed in the titration curve for each of the three types of ionisable hydrogens involved.

  14. Equilibrium position, kinetics, and reactor concepts for the adipyl-7-ADCA-hydrolysis process

    NARCIS (Netherlands)

    Schroën, C.G.P.H.; Wiel, van de S.; Kroon, P.J.; Vroom, de E.; Janssen, A.E.M.; Tramper, J.


    One of the building blocks of cephalosporin antibiotics is 7-amino-deacetoxycephalosporanic acid (7-ADCA). It is currently produced from penicillin G using an elaborate chemical ring-expansion step followed by an enzyme-catalyzed hydrolysis. However, 7-ADCA-like components can also be produced by

  15. Production of fermentable sugars by combined chemo-enzymatic hydrolysis of cellulosic material for bioethanol production

    Directory of Open Access Journals (Sweden)

    M. Idrees


    Full Text Available To change the recalcitrant nature of the lignocellulosic material for maximum hydrolysis yield, a comprehensive study was done by using sulphuric acid as an exclusive catalyst for the pretreatment process. The enzymatic digestibility of the biomass [Water Hyacinth: Eichhornia crassipes] after pretreatment was determined by measuring the hydrolysis yield of the pretreated material obtained from twenty four different pretreatment conditions. These included different concentrations of sulphuric acid (0.0, 1.0, 2.0 and 3.0%, at two different temperatures (108 and 121 ºC for different residence times (1.0, 2.0 and 3.0h.The highest reducing sugar yield (36.65 g/L from enzymatic hydrolysis was obtained when plant material was pretreated at 121 ºC for 1.0 h residence time using 3.0% (v/v sulphuric acid and at 1:10 (w/v solid to liquid ratio. The total reducing sugars obtained from the two-stage process (pretreatment + enzymatic hydrolysis was 69.6g/L. The resulting sugars were fermented into ethanol by using Saccharomyces cerevisiae. The ethanol yield from the enzymatic hydrolyzate was 95.2% of the theoretical yield (0.51g/g glucose, as determined by GS-MS, and nearly 100% since no reducing sugars were detected in the fermenting media by TLC and DNS analysis.

  16. Influence of starch source in the required hydrolysis time for the ...

    African Journals Online (AJOL)

    Influence of starch source in the required hydrolysis time for the production of maltodextrins with different dextrose equivalent. José Luis Montañez Soto, Luis Medina García, José Venegas González, Aurea Bernardino Nicanor, Leopoldo González Cruz ...

  17. The Reduction in the Rate of Hydrolysis of Diphenylbromomethane by the Common-Ion Effect (United States)

    Cameron-Holford, Richard; Ratneswaren, Tarini; Hughes, D. E. Peter


    Kinetic study on the hydrolysis of diphenylbromomethane does not provide clear-cut evidence for the mechanism of the reaction. The reduction in the rate of the reaction by added bromide ions suggested that the reaction took place by a S[subscript N]1 mechanism. A more detailed study of this common-ion effect, using conductivity to measure the rate…

  18. Steam pretreatment of spruce forest residues: optimal conditions for biogas production and enzymatic hydrolysis. (United States)

    Janzon, Ron; Schütt, Fokko; Oldenburg, Saskia; Fischer, Elmar; Körner, Ina; Saake, Bodo


    Steam refining of non-debarked spruce forest residues was investigated as pretreatment for enzymatic hydrolysis as well as for biogas production. Pretreatment conditions were varied in the range of 190-220 °C, 5-10 min and 0-3.7% SO₂ according to a statistical design. For both applications highest product yields were predicted at 220 °C and 2.4% SO₂, whereas the reaction time had only a minor influence. The conformity of the model results allows the conclusion that enzymatic hydrolysis is a suitable test method to evaluate the degradability of lignocellulosic biomass in the biogas process. In control experiments under optimal conditions the results of the model were verified. The yield of total monomeric carbohydrates after enzymatic hydrolysis was equivalent to 55% of all theoretically available polysaccharides. The corresponding biogas yield from the pretreated wood amounted to 304 mL/gODM. Furthermore, furans produced under optimal process conditions showed no inhibitory effect on biogas production. It can be concluded that steam refining opens the structure of wood, thus improving the enzymatic hydrolysis of the polysaccharides to fermentable monomeric sugars and subsequently enabling a higher and faster production of biogas. Anaerobic fermentation of pretreated wood is a serious alternative to alcoholic fermentation especially when low quality wood grades and residues are used. Anaerobic digestion should be further investigated in order to diversify the biorefinery options for lignocellulosic materials. Copyright © 2013 Elsevier Ltd. All rights reserved.

  19. Effects of acid-hydrolysis and hydroxypropylation on functional properties of sago starch. (United States)

    Fouladi, Elham; Mohammadi Nafchi, Abdorreza


    In this study, sago starch was hydrolyzed by 0.14M HCl for 6, 12, 18, and 24h, and then modified by propylene oxide at a concentration of 0-30% (v/w). The effects of hydrolysis and etherification on molecular weight distribution, physicochemical, rheological, and thermal properties of dually modified starch were estimated. Acid hydrolysis of starch decreased the molecular weight of starch especially amylopectin, but hydroxypropylation had no effect on the molecular weight distribution. The degree of Molar substitution (DS) of hydroxypropylated starch after acid hydrolysis ranged from 0.007 to 0.15. Dually modified starch with a DS higher than 0.1 was completely soluble in cold water at up to 25% concentration of the starch. This study shows that hydroxypropylation and hydrolysis have synergistic effects unlike individual modifications. Dually modified sago starch can be applied to dip-molding for food and pharmaceutical processing because of its high solubility and low tendency for retrogradation. Copyright © 2014 Elsevier B.V. All rights reserved.


    DEFF Research Database (Denmark)



    Ten new 2-substituted 2-methyl-2-oxy-4H-1,3-benzodioxin-4-ones, 4a-i and 6, have been synthesized and their rate constants of pseudo-first-order non-enzymatic hydrolysis determined. One of the compounds, 2-(2-acetoxybenzoyloxy)-2-methyl-4H-1,3-benzodioxin-4-one, 6, is a new anhydride of aspirin...

  1. Optimization of papain hydrolysis conditions for release of glycosaminoglycans from the chicken keel cartilage (United States)

    Le Vien, Nguyen Thi; Nguyen, Pham Bao; Cuong, Lam Duc; An, Trinh Thi Thua; Dao, Dong Thi Anh


    Glycosaminoglycans (GAGs) are natural biocompounds which join to construct cartilage tissuses, it can be extracted from cartilage of sharks, pigs, cows, chickens, etc. GAGs contain a Chondroitin sulfate (CS) content which is a supplement of functional food used for preventing and supporting treatment of arthritis and eye diseases. Therefore, the GAGs extraction from byproducts of the industry of cattle and poultry slaughter to identify the CS content by papain enzyme is necessary. In this study, the optimal hydrolysis conditions were obtained by response surface methodology (RSM). The independent variables were coded as: pH (x1), enzyme concentration (x2), incubation temperature (x3) and hydrolysis time (x4). The results of the analysis of variance (ANOVA) shown that the variables actively affected GAGs content. The optimal conditions of hydrolysis were derived at pH of 7.1, ratio of enzyme per substances of 0.62% w/wpo, temperature of 65°C and hydrolysis time of 230 minutes, GAGs content reached 14.3% of the dry matter of raw material. Analyzes by HPLC revealed that 56.17% of the dry preparations of GAGs were CS compound, were equivalent to 8.11% of the dry matter of chicken keel cartilage. Molecular weight of the dry preparations GAGs was 259.6 kDa. The dry preparations included the contents of moisture 12.2%, protein 8.42%, lipid 0%, ash 10.03% and extracted GAGs 69.35%.

  2. Enhanced anaerobic digestion performance via combined solids- and leachate-based hydrolysis reactor inoculation. (United States)

    Wilson, L Paige; Sharvelle, Sybil E; De Long, Susan K


    Suboptimal conditions in anaerobic digesters (e.g., presence of common inhibitors ammonia and salinity) limit waste hydrolysis and lead to unstable performance and process failures. Application of inhibitor-tolerant inocula improves hydrolysis, but approaches are needed to establish and maintain these desired waste-hydrolyzing bacteria in high-solids reactors. Herein, performance was compared for leach bed reactors (LBRs) seeded with unacclimated or acclimated inoculum (0-60% by mass) at start-up and over long-term operation. High quantities of inoculum (∼60%) increase waste hydrolysis and are beneficial at start-up or when inhibitors are increasing. After start-up (∼112days) with high inoculum quantities, leachate recirculation leads to accumulation of inhibitor-tolerant hydrolyzing bacteria in leachate. During long-term operation, low inoculum quantities (∼10%) effectively increase waste hydrolysis relative to without solids-derived inoculum. Molecular analyses indicated that combining digested solids with leachate-based inoculum doubles quantities of Bacteria contacting waste over a batch and supplies additional desirable phylotypes Bacteriodes and Clostridia. Copyright © 2016 Elsevier Ltd. All rights reserved.

  3. Selective enzymatic hydrolysis of C-terminal tert-butyl esters of peptides

    NARCIS (Netherlands)

    Eggen, I.F.; Boeriu, C.G.


    The present invention relates to a process for the selective enzymatic hydrolysis of C-terminal esters of peptide substrates in the synthesis of peptides, comprising hydrolysing C-terminal tert-butyl esters using the protease subtilisin. This process is useful in the production of protected or

  4. Selective enzymatic hydrolysis of C-terminal tert-butyl esters of peptides


    Eggen, I.F.; Boeriu, C.G.


    The present invention relates to a process for the selective enzymatic hydrolysis of C-terminal esters of peptide substrates in the synthesis of peptides, comprising hydrolysing C-terminal tert-butyl esters using the protease subtilisin. This process is useful in the production of protected or unprotected peptides.

  5. Surface chemical compositions and dispersity of starch nanocrystals formed by sulfuric and hydrochloric acid hydrolysis.

    Directory of Open Access Journals (Sweden)

    Benxi Wei

    Full Text Available Surface chemical compositions of starch nanocrystals (SNC prepared using sulfuric acid (H2SO4 and hydrochloric acid (HCl hydrolysis were analyzed by X-ray photoelectron spectroscopy (XPS and FT-IR. The results showed that carboxyl groups and sulfate esters were presented in SNC after hydrolysis with H2SO4, while no sulfate esters were detected in SNC during HCl-hydrolysis. TEM results showed that, compared to H2SO4-hydrolyzed sample, a wider size distribution of SNC prepared by HCl-hydrolysis were observed. Zeta-potentials were -23.1 and -5.02 mV for H2SO4- and HCl-hydrolyzed SNC suspensions at pH 6.5, respectively. Nevertheless, the zeta-potential values decreased to -32.3 and -10.2 mV as the dispersion pH was adjusted to 10.6. After placed 48 h at pH 10.6, zeta-potential increased to -24.1 mV for H2SO4-hydrolyzed SNC, while no change was detected for HCl-hydrolyzed one. The higher zeta-potential and relative small particle distribution of SNC caused more stable suspensions compared to HCl-hydrolyzed sample.

  6. Recovery and reuse of cellulase catalyst in an enzymatic cellulose hydrolysis process (United States)

    Woodward, J.


    A process for recovering cellulase from the hydrolysis of cellulose, and reusing it in subsequent hydrolyois procedures. The process utilizes a commercial adsorbent that efficiently removes cellulase from reaction products which can be easily removed by simple decantation. 1 fig., 4 tabs.

  7. Two-stage hydrolysis of invasive algal feedstock for ethanol fermentation. (United States)

    Wang, Xin; Liu, Xianhua; Wang, Guangyi


    The overall goal of this work was to develop a saccharification method for the production of third generation biofuel (i.e. bioethanol) using feedstock of the invasive marine macroalga Gracilaria salicornia. Under optimum conditions (120 °C and 2% sulfuric acid for 30 min), dilute acid hydrolysis of the homogenized invasive plants yielded a low concentration of glucose (4.1 mM or 4.3 g glucose/kg fresh algal biomass). However, two-stage hydrolysis of the homogenates (combination of dilute acid hydrolysis with enzymatic hydrolysis) produced 13.8 g of glucose from one kilogram of fresh algal feedstock. Batch fermentation analysis produced 79.1 g EtOH from one kilogram of dried invasive algal feedstock using the ethanologenic strain Escherichia coli KO11. Furthermore, ethanol production kinetics indicated that the invasive algal feedstock contained different types of sugar, including C(5) -sugar. This study represents the first report on third generation biofuel production from invasive macroalgae, suggesting that there is great potential for the production of renewable energy using marine invasive biomass. © 2011 Institute of Botany, Chinese Academy of Sciences.

  8. Hydrolysis and stability of thin pulsed plasma polymerised maleic anhydride coatings

    DEFF Research Database (Denmark)

    Drews, Joanna Maria; Launay, Héléne; Hansen, Charles M.


    were obtained at constant plasma power by adjusting the polymerisation time. The results show that the hydrolysis resistance of the modified layer is determined by the power used in the plasma polymerisation, while changes in the chemistry of the modified layer are insignificant....

  9. Enhanced enzymatic hydrolysis of kenaf core using irradiation and dilute acid (United States)

    Lee, Byoung-Min; Jeun, Joon-Pyo; Kang, Phil-Hyun


    This study was performed to determine the effect of electron beam dose and enzymatic hydrolysis time for production of sugar such as glucose and xylose. After kenaf core was exposed to an irradiation dose that ranged from 0 to 500 kGy, the irradiated kenaf core was treated with a 3% (v/v) sulfuric acid solution using an autoclave for 5 h at 120 °C. The pretreated kenaf core was subsequently subjected to enzymatic hydrolysis at 50 °C in a shaking water bath at 150 rpm for 12, 24, 48, and 72 h. The determined enzyme activity rates were 70 FPU (Celluclast 1.5 L) and 40 CBU (Novozyme-188). The crystallinity index decreased from 50.6% in a non-pretreated kenaf core to 27.7% in kenaf core that was subjected to the two-stage pretreatment at dose of 500 kGy. The sugar yield of the two-stage pretreated kenaf core increased with an increase in irradiation dose. The sugar yield after 72 h of enzymatic hydrolysis was 73.6% at its highest with an irradiation dose of 500 kGy. The enhancement of enzymatic hydrolysis by two-stage pretreatment was more effective than non- and single pretreatment (36.9%, 40.6% and 44.0% in non-pretreatment, electron beam and dilute acid, respectively).

  10. Analysis of the hydrolysis process in the formation of supergene zone mining areas

    Directory of Open Access Journals (Sweden)

    Sherstjuk N.P.


    Full Text Available The analysis of processes of hydrolysis of silicates and alumosilicat which arrive in a landscape from tails of enrichment of iron ore is carried out. Assessment and forecast formation of minerals zone hipcrhenezu iron deposits was carried out.

  11. Inside-out Z rings - constriction with and without GTP hydrolysis (United States)

    Osawa, Masaki; Erickson, Harold P.


    Summary The bacterial tubulin homolog FtsZ forms a ring-like structure called the Z ring that drives cytokinesis. We showed previously that FtsZ-YFP-mts, which has a short amphipathic helix (mts) on its C terminus that inserts into the membrane, can assemble contractile Z rings in tubular liposomes without any other protein. Here we study mts-FtsZ-YFP, where the membrane tether is switched to the opposite side of the protofilament. This assembled “inside-out” Z rings that wrapped around the outside surface of tubular liposomes. The inside-out Z rings were highly dynamic, and generated a constriction force that squeezed the tubular liposomes from outside. This is consistent with models where the constriction force is generated by curved protofilaments bending the membrane. We used this system to test how GTP hydrolysis by FtsZ is involved in Z-ring constriction. Without GTP hydrolysis, Z rings could still assemble and generate an initial constriction. However, the constriction quickly stopped, suggesting that Z rings became rigidly stabilized in the absence of GTP hydrolysis. We propose that remodeling of the Z ring, mediated by GTP hydrolysis and exchange of subunits, is necessary for the continuous constriction. PMID:21631604

  12. Identification of strong aggregating regions in soy glycinin upon enzymatic hydrolysis

    NARCIS (Netherlands)

    Kuipers, B.J.H.; Gruppen, H.


    Upon hydrolysis with chymotrypsin, soy glycinin has a strong tendency to aggregate. The regions of glycinin from which the aggregating peptides originate were identified by accumulative-quantitative peptide mapping. To this end, the aggregating peptides were further hydrolyzed with trypsin to obtain

  13. Lactose hydrolysis potential and thermal stability of commercial β-galactosidase in UHT and skimmed milk

    National Research Council Canada - National Science Library

    BOSSO, Alessandra; MORIOKA, Luiz Rodrigo Ito; SANTOS, Leandro Freire dos; SUGUIMOTO, Hélio Hiroshi


    ...(lyophilized) was investigated for its hydrolysis potential in lactose substrate, UHT milk, and skimmed milk at different concentrations (0.7; 1.0 and 1.5%), pH values (5.0; 6.0; 6.5 and 7.0), and temperature (30; 35; 40 and 55 ºC...


    Directory of Open Access Journals (Sweden)

    BĂLĂU MÎNDRU Tudorel


    Full Text Available The leather processing industry is known as an important source of environmental pollution, due to the large amount of waste generated. In this respect there are efforts concerning recovering and capitalizing of these wastes in order to subsequently use them in different applications. This work aimed to obtain collagen forms with convenient features starting from Chamois leather waste resulting from the buffing operation, by alkaline hydrolysis at different temperatures (60°C, 90°C, and 120°C and reaction time (4, 6 and 8 hours, in a solution of 30% NH4OH with a pH=11. The effects of the reaction conditions (temperature and reaction duration on the hydrolysis yield and molecular weight of the extracted polypeptides were investigated. The working version no. 8 (temperature = 1200C, and reaction time = 6h was chosen as optimal for obtaining a protein product suitable from the point of view of hydrolysis yield and molecular weight, respectively. For this sample no. 8, tests for checking physical-chemical characteristics were carried out, and also the influence of the hydrolysis upon certain structural changes caused by the reaction conditions was investigated by IR analysis. The reaction conditions led to obtaining polypeptide mixtures with different molecular weights and polydispersity that can be used as auxiliaries to obtain composite materials or as additives for new building materials.

  15. Acid Hydrolysis of Bromazepam Catalyzed by Micelles, Reverse Micelles, and Microemulsions

    Directory of Open Access Journals (Sweden)

    Ferdousi Begum


    Full Text Available Kinetics of the acid hydrolysis of bromazepam (Bz has been investigated in micelles, reverse micelles, and microemulcions of cetyltrimethylammonium bromide (CTAB by spectrophotometric method. The rate of the acid hydrolysis of Bz was found to be enhanced both below and above the critical micelle concentration (CMC of CTAB in aqueous solution. The pseudo-first-order rate constant (k′ shows an initial decrease for both low and high H+ concentrations. With further increase in [CTAB], at low [H+], the k′ attains an almost constant value, while, at high [H+], the k′ passes through a maximum and then decreases. The kinetic data for catalysis by micelles of CTAB was interpreted with the pseudophase ion exchange (PIE model. In CTAB/cyclohexane/1-butanol/water microemulsions, as the water to surfactant ratio (wo increases, the physicochemical properties and droplet sizes of microemulsions significantly change and distinct changes in reaction environment can be marked. The rate of the hydrolysis reaction exhibits excellent correlation with the physicochemical properties and droplet sizes of the microemulsions and reverse micelles of CTAB. At [H+] = 0.001 M, in reverse micelles and microemulsions of CTAB, the k′ of the acid hydrolysis of Bz decreases sharply followed by a slight increase with increasing wo.

  16. Production and effect of aldonic acids during enzymatic hydrolysis of lignocellulose at high dry matter content

    DEFF Research Database (Denmark)

    Cannella, David; Hsieh, Chia-Wen; Felby, Claus


    Abstract Background: The recent discovery of accessory proteins that boost cellulose hydrolysis has increased the economical and technical efficiency of processing cellulose to bioethanol. Oxidative enzymes (e.g. GH61) present in new commercial enzyme preparations have shown to increase cellulose...

  17. Eliminating inhibition of enzymatic hydrolysis by lignosulfonate in unwashed sulfite-pretreated aspen using metal salts (United States)

    Hao Liu; Junyong Zhu


    This study demonstrated the efficiency of Ca(II) and Mg(II) in removing inhibition of enzymatic hydrolysis by lignosulfonate through non-productive adsorption of enzymes. Adding 1 mmol/g cellulose of either metal salt restores approximately 65% of the activity lost when a pure cellulose/cellulase solution is spiked with lignosulfonate. Addition of either Ca(II) or Mg(...

  18. Effect of Acid mixtures on the Hydrolysis of Coconut Coir for ...

    African Journals Online (AJOL)


    ABSTRACT: In this study, coconut coir was hydrolysed to produce fermentable sugars using dilute nitric and acetic acid. The hydrolysis process was carried out according to a four variable Box-Behnken design which was used to develop a statistical model to describe the relationship between the concentration of ...

  19. Effect of Acid mixtures on the Hydrolysis of Coconut Coir for ...

    African Journals Online (AJOL)

    In this study, coconut coir was hydrolysed to produce fermentable sugars using dilute nitric and acetic acid. The hydrolysis process was carried out according to a four variable Box-Behnken design which was used to develop a statistical model to describe the relationship between the concentration of fermentable sugars ...

  20. Impact of the environmental conditions and substrate pre-treatment on whey protein hydrolysis: A review. (United States)

    Cheison, Seronei Chelulei; Kulozik, Ulrich


    Proteins in solution are subject to myriad forces stemming from interactions with each other as well as with the solvent media. The role of the environmental conditions, namely pH, temperature, ionic strength remains under-estimated yet it impacts protein conformations and consequently its interaction with, and susceptibility to, the enzyme. Enzymes, being proteins are also amenable to the environmental conditions because they are either activated or denatured depending on the choice of the conditions. Furthermore, enzyme specificity is restricted to a narrow regime of optimal conditions while opportunities outside the optimum conditions remain untapped. In addition, the composition of protein substrate (whether mixed or single purified) have been underestimated in previous studies. In addition, protein pre-treatment methods like heat denaturation prior to hydrolysis is a complex phenomenon whose progression is influenced by the environmental conditions including the presence or absence of sugars like lactose, ionic strength, purity of the protein, and the molecular structure of the mixed proteins particularly presence of free thiol groups. In this review, we revisit protein hydrolysis with a focus on the impact of the hydrolysis environment and show that preference of peptide bonds and/or one protein over another during hydrolysis is driven by the environmental conditions. Likewise, heat-denaturing is a process which is dependent on not only the environment but the presence or absence of other proteins.

  1. An integrated biohydrogen refinery: Synergy of photofermentation, extractive fermentation and hydrothermal hydrolysis of food wastes


    Redwood, Mark D.; Orozco, Rafael L.; Majewski, Artur J.; Macaskie, Lynne E


    An Integrated Biohydrogen Refinery (IBHR) and experimental net energy analysis are reported. The IBHR converts biomass to electricity using hydrothermal hydrolysis, extractive biohydrogen fermentation and photobiological hydrogen fermentation for electricity generation in a fuel cell. An extractive fermentation, developed previously, is applied to waste-derived substrates following hydrothermal pre treatment, achieving 83 99% biowaste destruction. The selective separation of organic acids fro...

  2. Surface chemical compositions and dispersity of starch nanocrystals formed by sulfuric and hydrochloric acid hydrolysis. (United States)

    Wei, Benxi; Xu, Xueming; Jin, Zhengyu; Tian, Yaoqi


    Surface chemical compositions of starch nanocrystals (SNC) prepared using sulfuric acid (H2SO4) and hydrochloric acid (HCl) hydrolysis were analyzed by X-ray photoelectron spectroscopy (XPS) and FT-IR. The results showed that carboxyl groups and sulfate esters were presented in SNC after hydrolysis with H2SO4, while no sulfate esters were detected in SNC during HCl-hydrolysis. TEM results showed that, compared to H2SO4-hydrolyzed sample, a wider size distribution of SNC prepared by HCl-hydrolysis were observed. Zeta-potentials were -23.1 and -5.02 mV for H2SO4- and HCl-hydrolyzed SNC suspensions at pH 6.5, respectively. Nevertheless, the zeta-potential values decreased to -32.3 and -10.2 mV as the dispersion pH was adjusted to 10.6. After placed 48 h at pH 10.6, zeta-potential increased to -24.1 mV for H2SO4-hydrolyzed SNC, while no change was detected for HCl-hydrolyzed one. The higher zeta-potential and relative small particle distribution of SNC caused more stable suspensions compared to HCl-hydrolyzed sample.

  3. Effects of thermo-chemical pre-treatment on anaerobic biodegradability and hydrolysis of lignocellulosic biomass

    NARCIS (Netherlands)

    Fernandes, T.; Klaasse Bos, G.J.; Zeeman, G.; Sanders, J.P.M.; Lier, van J.B.


    The effects of different thermo-chemical pre-treatment methods were determined on the biodegradability and hydrolysis rate of lignocellulosic biomass. Three plant species, hay, straw and bracken were thermo-chemically pre-treated with calcium hydroxide, ammonium carbonate and maleic acid. After

  4. The abnormal hydrolysis of 7-phosphanorbornenium salts: a case of phosphonium-phosphenium equivalence. (United States)

    Tian, Rongqiang; Liu, Hui; Duan, Zheng; Mathey, François


    The hydrolysis of a phosphanorbornenium triflate gives the expected tertiary phosphine oxide by cleavage of one P-C bond of the bridge in the presence of triethylamine but affords the secondary phosphine oxide by cleavage of the two P-C bonds of the bridge in the presence of alpha-picoline.

  5. Process development for gelatinisation and enzymatic hydrolysis of starch at high concentrations

    NARCIS (Netherlands)

    Baks, T.


    cum laude graduation (with distinction) Enzymatic hydrolysis of starch is encountered in day-to-day life for instance in the dishwasher during removal of stains with detergents or in our mouth during chewing of starch-based foods in the presence of saliva. The reaction is also important for the

  6. Enzymatic Cellulose Hydrolysis: Enzyme Reusability and Visualization of beta-Glucosidase Immobilized in Calcium Alginate

    DEFF Research Database (Denmark)

    Tsai, Chien Tai; Meyer, Anne S.


    The high cellulase enzyme dosages required for hydrolysis of cellulose is a major cost challenge in lignocellulosic ethanol production. One method to decrease the enzyme dosage and increase biocatalytic productivity is to re-use beta-glucosidase (BG) via immobilization. In the present research...

  7. Free energy diagram for the heterogeneous enzymatic hydrolysis of glycosidic bonds in cellulose

    DEFF Research Database (Denmark)

    Westh, Peter; Cruys-Bagger, Nicolaj; Sørensen, Trine Holst


    Kinetic and thermodynamic data has been analyzed according to transition state theory and a simplified reaction scheme for the enzymatic hydrolysis of insoluble cellulose. For the cellobiohydrolase Cel7A from Hypocrea jecorina (Tricoderma reesei) we were able to measure or collect relevant values...

  8. Cocaine benzoyl thioester: synthesis, kinetics of base hydrolysis, and application to the assay of cocaine esterases. (United States)

    Cashman, J R; Berkman, C E; Underiner, G; Kolly, C A; Hunter, A D


    The synthesis and characterization of diastereomers of cocaine benzoyl thioester is described. Allococaine benzoyl thioester and allopseudococaine benzoyl thioester were synthesized by the conjugate addition of p-methoxytolyl thiol to ecgonine methyl ester followed by debenzylation and benzoylation. The absolute structure of the hydrochloride salt of the major ecgonine p-methoxytolyl sulfide formed was determined by single-crystal diffraction analysis and used to establish the addition geometry. When placed in aqueous solution, the cocaine benzoyl thioester diastereomers hydrolyzed to give thioecgonine methyl ester. The rate of cocaine benzoyl thioester hydrolysis was carefully investigated spectrophotometrically by using the Ellman reagent. At neutral pH, the hydrolysis of the diastereomers was found to proceed at detectable rates. Upon increasing pH, the rate of hydrolysis of cocaine benzoyl thioester diastereomers was increased and the reaction was catalyzed by basic buffer species. In addition to defining the kinetics of hydrolysis in aqueous solution, cocaine benzoyl thioester was utilized as a highly efficient method to monitor the activity of cholinesterases and pig liver esterase. Use of cocaine benzoyl thioester represents a rapid and sensitive way to screen for cocaine esterase activity.

  9. Effective of Microwave-KOH Pretreatment on Enzymatic Hydrolysis of Bamboo (United States)

    Zhiqiang Li; Zehui Jiang; Yan Yu; Zhiyong Cai


    Bamboo, with its advantages of fast growth, short renovation, easy propagation and rich in cellulose and hemicellulose, is a potential feedstock for bioethanol or other biofuels production. The objective of this study was to examine the fea- sibility of microwave assistant KOH pretreatments to enhance enzymatic hydrolysis of bamboo. Pretreatment was car- ried out by...

  10. Oleuropein hydrolysis in natural green olives: Importance of the endogenous enzymes. (United States)

    Ramírez, Eva; Brenes, Manuel; García, Pedro; Medina, Eduardo; Romero, Concepción


    The bitter taste of olives is mainly caused by the phenolic compound named oleuropein and the mechanism of its hydrolysis during the processing of natural green olives was studied. First, a rapid chemical hydrolysis of oleuropein takes place at a high temperature of 40°C and at a low pH value of 2.8, but the chemical hydrolysis of the bitter compound is slow at the common range of pH for these olives (3.8-4.2). However, decarboxymethyl elenolic acid linked to hydroxytyrosol and hydroxytyrosol have been found in a high concentration during the elaboration of natural green olives. When olives were heated at 90°C for 10min before brining, these compounds are not formed. Hence, the debittering process in natural green olives is due to the activity of β-glucosidase and esterase during the first months of storage and then a slow chemical hydrolysis of oleuropein happens throughout storage time. Copyright © 2016 Elsevier Ltd. All rights reserved.

  11. A discretized model for enzymatic hydrolysis of cellulose in a fed-batch process. (United States)

    Tervasmäki, Petri; Sotaniemi, Ville; Kangas, Jani; Taskila, Sanna; Ojamo, Heikki; Tanskanen, Juha


    In the enzymatic hydrolysis of cellulose, several phenomena have been proposed to cause a decrease in the reaction rate with increasing conversion. The importance of each phenomenon is difficult to distinguish from batch hydrolysis data. Thus, kinetic models for the enzymatic hydrolysis of cellulose often suffer from poor parameter identifiability. This work presents a model that is applicable to fed-batch hydrolysis by discretizing the substrate based on the feeding time. Different scenarios are tested to explain the observed decrease in reaction rate with increasing conversion, and comprehensive assessment of the parameter sensitivities is carried out. The proposed model performed well in the broad range of experimental conditions used in this study and when compared to literature data. Furthermore, the use of data from fed-batch experiments and discretization of the model substrate to populations was found to be very informative when assessing the importance of the rate-decreasing phenomena in the model. Copyright © 2016 Elsevier Ltd. All rights reserved.

  12. Coal liquefaction by base-catalyzed hydrolysis with CO.sub.2 capture (United States)

    Xiao, Xin


    The one-step hydrolysis of diverse biomaterials including coal, cellulose materials such as lumber and forestry waste, non-food crop waste, lignin, vegetable oils, animal fats and other source materials used for biofuels under mild processing conditions which results in the formation of a liquid fuel product along with the recovery of a high purity CO.sub.2 product is provided.

  13. Effect of pretreatment on the enzymatic hydrolysis of kitchen waste for xanthan production. (United States)

    Li, Panyu; Zeng, Yu; Xie, Yi; Li, Xiang; Kang, Yan; Wang, Yabo; Xie, Tonghui; Zhang, Yongkui


    The study was carried out to gain insight into the effect of pretreatment on enzymatic hydrolysis of kitchen waste (KW) for xanthan fermentation. Herein, various pretreatments were applied and it was found that chemical pretreatment had positive effect on the following enzymatic or overall hydrolysis process. The highest reducing sugar concentration was obtained as 51.87g/L from 2% HCl (90°C) pretreated sample, while the Kjeldahl nitrogen (KDN) concentration was 7.79g/L. Kinetic study showed that first order kinetic model was suitable to describe the enzymatic hydrolysis process. The obtained kitchen waste hydrolysate (KWH) was successfully applied for xanthan fermentation. Xanthan concentration reached 4.09-6.46g/L when KWH with 2% HCl (90°C) pretreatment was applied as medium. In comparison, a xanthan concentration of 3.25-5.57g/L was obtained from KWH without pretreatment. Therefore, pretreatment of KW using diluted acid is favorable for the overall hydrolysis process and effective for xanthan fermentation. Copyright © 2016 Elsevier Ltd. All rights reserved.

  14. Investigation of Unexpected Reaction Intermediates in the Alkaline Hydrolysis of Methyl 3,5-Dinitrobenzoate (United States)

    Silva, Clesia C.; Silva, Ricardo O.; Navarro, Daniela M. A. F.; Navarro, Marcelo


    An experimental project aimed at identifying stable reaction intermediates is described. Initially, the studied reaction appears to involve the simple hydrolysis, by aqueous sodium hydroxide, of methyl 3,5-dinitrobenzoate dissolved in dimethyl sulfoxide. On mixing the substrates, however, the reaction mixture unexpectedly turns an intense red in…

  15. Cation Transport Coupled to ATP Hydrolysis by the (Na, K)-ATPase: An Integrated, Animated Model (United States)

    Leone, Francisco A.; Furriel, Rosa P. M.; McNamara, John C.; Horisberger, Jean D.; Borin, Ivana A.


    An Adobe[R] animation is presented for use in undergraduate Biochemistry courses, illustrating the mechanism of Na[superscript +] and K[superscript +] translocation coupled to ATP hydrolysis by the (Na, K)-ATPase, a P[subscript 2c]-type ATPase, or ATP-powered ion pump that actively translocates cations across plasma membranes. The enzyme is also…

  16. Enzymatic hydrolysis of loblolly pine: effects of cellulose crystallinity and delignification (United States)

    Umesh P. Agarwal; J.Y. Zhu; Sally A. Ralph


    Hydrolysis experiments with commercial cellulases have been performed to understand the effects of cell wall crystallinity and lignin on the process. In the focus of the paper are loblolly pine wood samples, which were systematically delignified and partly ball-milled, and, for comparison, Whatman CC31 cellulose samples with different crystallinities. In pure cellulose...

  17. Determination of kinetics and heat of hydrolysis for non-homogenous substrate by isothermal calorimetry. (United States)

    Tafoukt, D; Soric, A; Sigoillot, J-C; Ferrasse, J-H


    The competitiveness of the second-generation bioethanol by biotechnological process requires an effective and quantitative control of biochemical reactions. In this study, the potential of isothermal calorimetry technique to measure heat and kinetics of a non-homogeneous substrate enzymatic hydrolysis is intended. Using this technique, optimum temperature of the enzymes used for lignocellulosic molecules hydrolysis was determined. Thus, the amount of substrate-to-enzyme ratio was highlighted as an important parameter of the hydrolysis yield. Furthermore, a new enzymes' cocktail efficiency consisting of a mix of cellulases and cellobiose dehydrogenase (CDH) was qualified by this technique. The results showed that this cocktail allowed the production of a high amount of gluconic acid that could improve the attractiveness of these second-generation biofuels. From the set of experiments, the hydrolysis heat of wheat straw was derived and a meaningful value of -32.2 ± 3.2 J g(-1) (gram reducing sugars product) is calculated. Then, isothermal measurements were used to determine kinetic constants of the cellulases and CDH mix on wheat straw. Results showed that this enzyme cocktail has an optimal rate at 45 °C in the range of temperatures tested (40-55 °C).

  18. Enhancement of high-solids enzymatic hydrolysis of corncob residues by bisulfite pretreatment for biorefinery. (United States)

    Xing, Yang; Bu, Lingxi; Zheng, Tianran; Liu, Shijie; Jiang, Jianxin


    Co-production of glucose, furfural and other green materials based on a lignocellulosic biorefinery is a promising way to realize the commercial application of corncob residues. An effective process was developed for glucose production using low temperature bisulfite pretreatment and high-solids enzymatic hydrolysis. Corncob residues from furfural production (FRs) were pretreated with 0.1g NaHSO3/g dry substrate at 100°C for 3h. Lignin was sulfonated and sulfonic groups were produced during pretreatment, which resulted in decreasing the zeta potential of the samples. Compared with raw material, bisulfite pretreatment of FRs increased the glucose yield from 18.6 to 99.45% after 72h hydrolysis at a solids loading of 12.5%. The hydrolysis residues showed a relatively high thermal stability and concentrated high derivatives. Direct pretreatment followed by enzymatic hydrolysis is an environmentally-friendly and economically-feasible method for the production of glucose and high-purity lignin, which could be further converted into high-value products. Copyright © 2016 Elsevier Ltd. All rights reserved.

  19. A vertical ball mill as a new reactor design for biomass hydrolysis and fermentation process

    DEFF Research Database (Denmark)

    de Assis Castro, Rafael Cunha; Mussatto, Solange I.; Conceicao Roberto, Inês


    A vertical ball mill (VBM) reactor was evaluated for use in biomass conversion processes. The effects of agitation speed (100–200 rpm), number of glass spheres (0–30 units) and temperature (40–46 °C) on enzymatic hydrolysis of rice straw and on glucose fermentation by a thermotolerant Kluyveromyc...

  20. Improved toluidine blue-DNA agar for detection of DNA hydrolysis by campylobacters.


    Lior, H.; PATEL, A.


    Our improved toluidine blue-DNA agar was compared with methyl green DNase test agar for the detection of DNA hydrolysis by campylobacters. The toluidine blue-DNA agar gave clear-cut positive and negative reactions with campylobacter strains belonging to several species.