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Sample records for enzymatic interesterification evaluation

  1. Modification of margarine fats by enzymatic interesterification:

    DEFF Research Database (Denmark)

    Zhang, Hong; Pedersen, Lars Saaby; Kristensen, Dorther

    2004-01-01

    to the equilibrium state, and (iii) the reaction rate constant value (k). SFC0 and ΔSFC were related to only the types of blends and the blend ratios. The rate constant k was related to lipase activity on a given oil blend. Evaluation of the model was carried out with two groups of oil blends, i.e., palm stearin/coconut...... oil in weight ratios of 90:10, 80:20, and 70:30, and soybean oil/fully hydrogenated soybean oil in weight ratios of 80:20, 65:35, and 50:50. Correlation coefficients higher than 0.99 between the experimental and predicted values were observed for SFC at temperatures above 30°C. The model is useful...... for predicting changes in the SFC during lipase-catalyzed interesterification with a selected group of oil blends. It also can be used to control the process when particular SFC values are targeted....

  2. Production of specific-structured lipids by enzymatic interesterification in a pilot continuous enzyme bed reactor

    DEFF Research Database (Denmark)

    Xu, Xuebing; Balchen, Steen; Høy, Carl-Erik

    1998-01-01

    Production of specific-structured lipids (interesterified lipids with a specific structure) by enzymatic interesterification was carried out in a continuous enzyme bed pilot scale reactor. Commercial immobilized lipase (Lipozyme IM) was used and investigations of acyl migration, pressure drop...

  3. Structured Triacylglycerol of Palm-based Margarine Fat by Enzymatic Interesterification

    DEFF Research Database (Denmark)

    Ibrahim, Nuzul Amri Bin

    2008-01-01

    blends containing palm stearin (PS) (60 to 90%), palm kernel oil (PKO) (10 to 40%) and fish oil (FO) (0 to 10%) were interesterified by Lipozyme TL IM (Thermomyces lanuginosa) using a continuous packed bed reactor (PBR). FO in the blend had a similar effect as PKO on solid fat content (SFC...... PBR for enzymatic interesterification of sunflower oil and fish oil blend. The pre-column functions as a protector of the enzyme bed to prolong the lifespan of the enzyme. Deactivation kinetic of enzymes subjected to interesterification using a pre-column was compared against without a pre......-column. The deactivation rate of enzyme bed without a pre-column was 7.4 times faster than the one with a pre-column, where the half-life were 8 and 58 days respectively....

  4. Human Milk Fat Substitute Produced by Enzymatic Interesterification of Vegetable Oil Blend

    Directory of Open Access Journals (Sweden)

    Semra Turan

    2007-01-01

    Full Text Available Palm oil, palm kernel oil, olive oil, sunflower oil, and marine oil blend, formulated in the mass ratio of 4.0:3.5:1.0:1.5:0.2, was subjected to interesterification catalyzed by lipase from Thermomyces lanuginosa (Lipozyme® TL IM for obtaining a product that contains similar triacylglycerol (TAG structure to that of human milk fat (HMF. Reactions were carried out in a double jacketed glass vessel equipped with magnetic stirrer at 60 °C for 2, 4, 6, 8, 12 and 24 h. The blend was analyzed for fatty acid composition of both total fatty acids and those at the sn-2 position after pancreatic lipase hydrolysis. After interesterification, TAGs were purified by thin layer chromatography and TAG species were determined according to the carbon number (CN by high-temperature gas chromatography. Enzymatic interesterification generated significant differences for all TAG species from CN30 to CN54. Concentrations of some TAG species (CN30, 32, 34, 36, 38, 50, 52 and 54 decreased, while some (CN40 to 48 increased after 24 h. TAG species with higher CN reached maximum levels at the end of 6 h of reaction time. The predominant TAGs of the reaction product after 24 h were CN46, 48, 50, 52 and 54 with ratios of 13.8, 18.2, 13.9, 17.8, and 12.1 %, respectively. These TAG species contain mainly 1,3-diunsaturated-2-saturated structure, like HMF.

  5. Enzymatic interesterification of vegetable oil/ fish oil blend for margarine production

    DEFF Research Database (Denmark)

    Ibrahim, Nuzul Amri Bin; Xu, Xuebing

    the desired properties. In this study, palm stearin (PS), palm kernel oil (PKO) and fish oil (FO) are blended and modified by enzymatic interesterification. PS functioned as the hard stock, PKO as the soft oil and FO as a source for eicosapentaenoic acid (EPA)/ docosahexaenoic acid (DHA). The purpose...... cause the product to be susceptible to oxidation due to the presence of high content of polyunsaturated fatty acids. Furthermore, FO could also influence the melting properties of the product. Therefore, in addition to determining the fatty acid position on the glycerol backbone, it is also pertinent...

  6. Functional palm oil-based margarine by enzymatic interesterification

    DEFF Research Database (Denmark)

    Ibrahim, Nuzul Amri Bin; Xu, Xuebing

    Palm stearin, palm kernel and fish oils were blended to a various composition ratios and enzymatically interesterified by Lipozyme TL IM lipase (Thermomyces lanuginosa) using a continuous packed bed reactor. The ratio of the oils ranged from 60-90%, 10-40% and 0-10% respectively. The enzyme was a...

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

    DEFF Research Database (Denmark)

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

    2008-01-01

    greater than 100% over the theoretical value when the reaction proceeds for 2 h. The co-immobilization action of the carrier of the immobilized lipases towards the free lipase was proposed as being one of the reasons leading to the synergistic effect and this has been experimentally verified by a reaction......Enzymatic interesterification of palm stearin with coconut oil was conducted by applying a dual lipase system in comparison with individual lipase-catalyzed reactions. The results indicated that a synergistic effect occurred for many lipase combinations, but largely depending on the lipase species...... mixed and their ratios. The combination of Lipozyme TL IM and RM IM was found to generate a positive synergistic action at all test mixing ratios. Only equivalent amount mixtures of Lipozyme TL IM with Novozym 435 or Lipozyme RM IM with Novozym 435 produced a significant synergistic effect as well...

  8. Comparison of Chemical and Enzymatic Interesterification of Fully Hydrogenated Soybean Oil and Walnut Oil to Produce a Fat Base with Adequate Nutritional and Physical Characteristics

    Directory of Open Access Journals (Sweden)

    Mariel Farfán

    2015-01-01

    Full Text Available The optimal physical, chemical and nutritional properties of natural lipids depend on the structure and composition of triacylglycerols. However, they are not always mutually compatible. Lipid modification is a good way to give them specific functionalities, increase their oxidative stability, or improve their nutritional value. As such, chemical and enzymatic interesterification may be used to modify them and produce structured lipids. In accordance, the aim of this study is to compare chemical and enzymatic interesterifi cation of binary blends of fully hydrogenated soybean oil and walnut oil, using sodium methoxide or Lipozyme TL IM, respectively, to produce a fat base with adequate nutritional and physical characteristics. Three different mass ratios of fully hydrogenated soybean oil and walnut oil blends (20:80, 40:60 and 60:40 were interesterified and evaluated. Total interesterification was determined by the stabilization of the solid fat content. Chemical reaction of the 20:80 blend was completed in 10 min and of the 40:60 and 60:40 blends in 15 min. Enzymatically interesterified blends were stabilized in 120 min at all of the mass ratios. Complete interesterification significantly reduced the solid fat content of the blends at any composition. Chemical and enzymatically interesterified fully hydrogenated blend of soybean and walnut oil at mass ratio of 40:60 showed the plastic curve of an all-purpose-type shortening rich in polyunsaturated fatty acids, with a high linolenic acid (C18:3n3 content and with zero trans-fatty acids.

  9. Assessing the reaction conditions to mediate the milkfat-soybean oil enzymatic interesterification

    Directory of Open Access Journals (Sweden)

    Ariela Veloso de Paula

    Full Text Available Summary A food grade lipase from Rhizopus oryzae immobilized on a hybrid polysiloxane-polyvinyl alcohol matrix (SiO2-PVA was used as the biocatalyst to mediate the interesterification reactions of a blend containing 65% milkfat and 35% soybean oil. All the reactions occurred in an inert nitrogen atmosphere in cylindrical glass reactors (80 mL with 40 g of the milkfat-soybean oil blend. The influence of the following variables was evaluated: biocatalyst loading (250-1500 activity units per gram of blend, biocatalyst moisture content (5-20%, temperature (45-60 °C and incubation time (2-48 h. The reactions were monitored by determining the free fatty acid content, triacylglycerol (TAGs composition in carbon species, and the consistency of the interesterified (IE products. The reaction conditions were set based on the parameters that provided a high interesterification yield and good consistency of the final product within the ideal range (200 to 800 gf cm-2. Hence the best results were obtained using a biocatalyst loading of 500 U g-1 of blend with 10% moisture content at 45 °C for 4 h. Under these conditions the consistency of the interesterified product was 539.7 ± 38 gf cm-2. The results demonstrated the potential of the immobilized lipase to alter the TAGs profile of the milkfat-soybean oil blend, allowing for the production of structured lipids.

  10. PEMBUATAN BAHAN BAKU SPREADS KAYA KAROTEN DARI MINYAK SAWIT MERAH MELALUI INTERESTERIFIKASI ENZIMATIK MENGGUNAKAN REAKTOR BATCH [Preparation of Red Palm Oil Based-Spreads Stock Rich in Carotene Through Enzymatic Interesterification in Batch-type Reactor

    Directory of Open Access Journals (Sweden)

    Nur Wulandari1,2

    2012-12-01

    Full Text Available Enzymatic interesterification of red palm oil (a mixture of red palm olein/RPO and red palm stearin/RPS in 1:1 weight ratio and coconut oil (CNO blends of varying proportions using a non-specific immobilized Candida antartica lipase (Novozyme 435 was studied for the preparation of spread stock. The interesterification reaction was held in a batch-type reactor. Two substrate blends were chosen for the production of spread stock i.e. 77.5:22,5 and 82.5:17.5 (RPO/RPS:CNO, by weight through enzymatic interesterification in three different reaction times (2, 4, and 6 hours. The interesterification reactions were conducted at 60°C, 200 rpm agitation speed and 10% of Novozyme 435. The interesterified products were evaluated for their physical characteristics (slip melting point or SMP and solid fat content or SFC and chemical characteristics (carotene retention, moisture content, and free fatty acid/FFA content. All of the interesterified products had lower SFC and SMP as compared to the initial blends. The SMP and SFC increased in longer reaction times. The SMP ranged from 30.8°C to 34.9°C. The carotene retention ranged from 74.80% to 81.08%, while the moisture content and FFA content increased in longer reaction times. The interesterified products had desirable physical properties for possible use as a spread stock rich in carotene.

  11. Incorporation of medium chain fatty acids into fish oil triglycerides by chemical and enzymatic interesterification

    Directory of Open Access Journals (Sweden)

    2009-06-01

    Full Text Available Structured triglycerides (STs containing both medium-chain fatty acids (MCFA and polyunsaturated fatty acids (PUFA in the same molecule offer nutritional and therapeutic benefits. The aim of this work was to establish the incorporation of MCFA into fish oil triglycerides (TAGs, while maintaining substantial levels of docosahexaenoic and eicosapentaenoic acids. The effects of different acyl donors (capric acid methyl ester/MeC10 or medium chain triglyceride/TCM and of the catalyst (chemical or enzymatic on the fatty acid composition of the reaction product were studied. The fatty acid composition of the fish oil TAG was modified after interesterification to contain MCFA, and it depended on the catalyst and on the substrates. Thermograms obtained by Differential Scanning Calorimetry (DSC showed that interesterification promoted noteworthy changes in the melting profile of the samples. STs of clinical nutrition interest containing both EPA and DHA obtained from fish oil along with MCFA were successfully produced.

    Triglicéridos estructurados (SL conteniendo ácidos grasos de cadena media (MCFA y ácidos grasos poliinsaturados (PUFA en la misma molécula de glicerol tienen ventajas nutricionales y terapéuticas. Se establece la incorporación de MCFA a los triglicéridos (TAGs de aceite de pescado, conservando un contenido considerable de ácidos docosahexaenóico (DHA y eicosapentaenóico (EPA. El efecto de diferentes acil donadores (éster metílico de ácido cáprico/MeC10 o triglicéridos de cadena media/TCM y de catalizador (químico o enzimático sobre la composición del producto de las reacciones fue estudiado. La composición de ácidos grasos de los TAGs del aceite de pescado fue modificada después de las reacciones para contener MCFA y dependió del catalizador y de los substratos. Los termogramas obtenidos por Calorimetría Diferencial de Barrido (DSC indicaron que la interesterificación provocó alteraciones considerables de

  12. Enzymatic interesterification of butterfat with rapeseed oil in a continuous packed bed reactor

    DEFF Research Database (Denmark)

    Rønne, Torben Harald; Yang, Tiankui; Mu, Huiling

    2005-01-01

    , whereafter it dramatically decreased over the next 10 days to an activity level of 40%. In general, the study shows no significant difference for butterfat interesterification in terms of enzyme behavior from normal vegetable oils and fats even though it contains short-chain fatty acids and cholesterol......Lipase-catalyzed interesterification of butterfat blended with rapeseed oil (70/30, w/w) was investigated both in batch and in continuous reactions. Six commercially available immobilized lipases were screened in batch experiments, and the lipases, Lipozyme TL IM and Lipozyme RM IM, were chosen...

  13. INTERESTERIFIKASI ENZIMATIS MINYAK IKAN DENGAN ASAM LAURAT UNTUK SINTESIS LIPID TERSTRUKTUR [Enzymatic Interesterification of Fish Oil with Lauric Acid for the Synthesis of Structured Lipid

    Directory of Open Access Journals (Sweden)

    Edy Subroto1

    2008-12-01

    Full Text Available Structured lipid (SL containing of medium chain fatty acid (MCFA at outer position and polyunsaturated fatty acid (PUFA at sn-2 position has superior dietary and absorption characteristics. The most methods for the enzymatic synthesis of SL were through two steps process, so that it was inefficient. Caprilic acid was usually used as a source of MCFA. In this research, SL was synthesized by enzymatic interesterification between fish oil and lauric acid. The specific lipase from Mucor miehei was used as catalyzed. Factors, such as the incubation time, substrate mole ratio, and reaction temperature were evaluated. The incorporation and the position of lauric acid on glycerol backbone and glyceride profile were determined. The results showed that SL containing of lauric acid at the outer position and PUFA at sn-2 was successfully synthesized, and it was done through one step process. From regiospecific determination, it showed that the position of lauric acid incorporation was only at the sn-1 and sn-3. Only 0.87% of lauric acid was incorporated at the sn-2. The optimum time and temperature of the reaction, and the substrate mole ratio were 12 h, 50C and 1:10, respectively, in which the incorporation of lauric acid was 62.8% (mol. Glyceride profile was affected by incubation time, substrate mole ratio and reaction temperature. Triglyceride concentration decreased with an increase in the incubation time (> 12 h. In contrast, the diglyceride concentration increased at longer incubation time (> 12 h. Beside, triglyceride concentration increased with an increase in substrate mole ratio to 1:10, but it decreased when mole ratio of substrate was 1:15. At higher temperature (50C, triglyceride decreased with an increase in the reaction temperature. In summary, the SL was successfully synthesized by the interesterification of fish oil and lauric acid using specific lipase of Mucor miehei.

  14. INTERESTERIFIKASI ENZIMATIS MINYAK IKAN DENGAN ASAM LAURAT UNTUK SINTESIS LIPID TERSTRUKTUR [Enzymatic Interesterification of Fish Oil with Lauric Acid for the Synthesis of Structured Lipid

    OpenAIRE

    Edy Subroto1); Chusnul Hidayat2); Supriyadi2)

    2008-01-01

    Structured lipid (SL) containing of medium chain fatty acid (MCFA) at outer position and polyunsaturated fatty acid (PUFA) at sn-2 position has superior dietary and absorption characteristics. The most methods for the enzymatic synthesis of SL were through two steps process, so that it was inefficient. Caprilic acid was usually used as a source of MCFA. In this research, SL was synthesized by enzymatic interesterification between fish oil and lauric acid. The specific lipase from Mucor miehei...

  15. Production of structured lipids: acyl migration during enzymatic interesterification and downstream processing

    DEFF Research Database (Denmark)

    Xu, Xuebing

    1997-01-01

    Production of structured lipids by lipase-catalyzed interesterification attracts great interests recently. Structured lipids are defined, in this article, as triacylglycerols which contain both medium or short chain fatty acids and long chain fatty acids, each groups locating specifically in the sn......-2 position or sn-1,3 positions of glycerol backbone. These kinds of lipids are reported to be promising for both enteral and parenteral nutrition. However, acyl migration occurs in the reaction stage and downstream purification process. This side-reaction causes by-products which are harmful...

  16. Pilot batch production of cocoa butter-like fats from chinese vegetable tallow by enzymatic interesterification

    DEFF Research Database (Denmark)

    Xu, Xuebing; Hu, X.; Balchen, Steen

    1997-01-01

    -8%. And about 90% of the present PPP and PPSt triglycerides were separated from the product. Under above parameters, the final pilot products had similar compositions to those of cocoa butter. In this research, IPPL showed initial interesterification activity at the similar level as Lipozyme IM from Novo......There is a long term interest of lipase applications in lipid modifications because of the inherent advantages over chemical methods such as more specific reactions involved, less energy used, moderate reaction conditions and so on. In this work, cocoa butter-like fats (CBF) were produced using....../w) 1.3-1.7/1, hexane/substrates(v/w): 1.0:1-1.2:1. Based on the pilot batch plant, the total process is following: substrates, drying, reaction, filtration, fractionation, neutralisation, de-solventization and drying, and product. The fractionation process reduced the free fatty acid content to 5...

  17. INTERESTERIFIKASI ENZIMATIS PALM STEARIN DAN MINYAK IKAN LEMURU UNTUK MEMBUAT LEMAK MARGARIN [Enzymatic Interesterification of Palm Stearin and Sardine oil to Produce Margarine-fat

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    Pudji Hastuti

    2003-04-01

    Full Text Available Enzymatic interesterification of Palm Stearin (PS and Sardine Oil (SO as source of Eicosa Pentaenoic Acid (EPA and Docosa Hexaenoic Acid (DHA have been of interest to modify the physical properties of the triglyceride. An attempt to enzymatic-restructure PS and SO to form Structured Lipid (SL which is suitable for margarine was investigated using immobilized lipase from Rhizomucor miehei and that from Candida antartica. The effect of reaction time course, ratio of PS/SO and ratio of enzyme/substrate were studied in the present study. At the end of interesterification, the enzyme was filtered from the reaction mixture through a filter paper. The Solid Fat Index (SFI was determined by dillatometry. The Slip Melting Point (SMP was determined by capillary tube method. Both of interesterification catalyzed by immobilized sn 1,3 specific lipase from R.miehei,and non specific lipase from C.antartica were found to decrease the SFI value at 10; 21.1 and 33.3°C. The SMP value was decrease from 58-50°C to 37-39°C. The change of these parameters were slightly faster in the reaction which catalyzed by lipase from R miehei than lipase from C.antartica . The more the utilization of the enzyme the faster the change were occurred, especially the increase of enzyme utilization from 2.5% to 5%, which decrease the SFI value at 33,30C. The decrease of the PS/SO ratio resulted in the decrease of SFI and SMP values. It was found that the most suitable SFI and SMP value for margarine fat is the SL formed by carrying out the enzymatic-interesterification of PS/SO with the ratio of 40/60 using enzyme 2.5% of the total fat, for 8 hours at 60°C.

  18. Influence of enzymatic and chemical interesterification on crystallisation properties of refined, bleached and deodourised (RBD) palm oil and RBD palm kernel oil blends.

    Science.gov (United States)

    Norizzah, Abd Rashid; Nur Azimah, Kamarulzaman; Zaliha, Omar

    2018-04-01

    Interesterification reaction involves rearrangement of the fatty acid radicals on the glycerol backbone, either randomly (chemical interesterification) or regioselectivity (enzymatic interesterification). Refined, bleached and deodourised palm oil (RBDPO) and palm kernel oil (RBDPKO) were blended in ratios from 25:75 to 75:25 (wt/wt). All blends were subjected to enzymatic (EI) and chemical interesterification (CI) using Lipozyme TL IM (4% w/w) and sodium methoxide (0.2% m/m) as the catalysts, respectively. The effect of EI and CI on the triacylglycerol (TAG) composition, thermal behaviour, polymorphism, crystal morphology and crystallisation kinetics were studied. The aim of this research is to characterise the nature of crystals in food product for certain desired structure. The crystallisation behaviour discussed in this study involves microstructure (PLM), polymorphism (XRD), thermal properties and crystallisation kinetics by DSC. The alteration in TAG composition was greater after CI as compared to EI with the reduction of LaLaLa (from 11.00% to 5.15%) and POO (from 14.28% to 4.87%). The DSC complete melting and crystallisation temperature of blend with 75% PO increased after CI, from 39.58 °C to 41.67 °C and from -30.84 °C to -28.33 °C, respectively. EI contributed to finer crystals than CI. However, the β' and β polymorph mixture and crystallisation kinetics (n = 2) of PO-PKO blends did not change after CI and EI. The knowledge on controlling crystallisation of RBDPO and RBDPKO blends is vital for proper processing condition like margarine production. Copyright © 2018 Elsevier Ltd. All rights reserved.

  19. Production of margarine fats by enzymatic interesterification with silica-granulated Thermomyces lanuginosa lipase in a large-scale study

    DEFF Research Database (Denmark)

    Zhang, Hong; Xu, Xuebing; Nilsson, Jörgen

    2001-01-01

    Interesterification of a blend of palm stearin and coconut oil (75:25, w/w), catalyzed by an immobilized Thermomyces lanuginosa lipase by silica granulation, Lipozyme TL IM, was studied for production of margarine fats in a 1- or 300-kg pilot-scale batch-stirred tank reactor. Parameters...

  20. Production of specific-structured lipids by enzymatic interesterification: elucidation of acyl migration by response surface design

    DEFF Research Database (Denmark)

    Xu, Xuebing; Skands, Anja; Høy, Carl-Erik

    1998-01-01

    Production of specific-structured lipids (SSL) by lipase-catalyzed interesterification has been attracting more and more attention recently. However, it was found that acyl migration occurs during the reaction and causes the production of by-products. In this paper, the elucidation of acyl...

  1. Production of specifically structured lipids by enzymatic interesterification in a pilot enzyme bed reactor: process optimization by response surface methodology

    DEFF Research Database (Denmark)

    Xu, Xuebing; Mu, Huiling; Høy, Carl-Erik

    1999-01-01

    Pilot production of specifically structured lipids by Lipozyme IM-catalyzed interesterification was carried out in a continuous enzyme bed reactor without the use of solvent. Medium chain triacylglycerols and oleic acid were used as model substrates. Response surface methodology was applied...... and the production of mono-incorporated and di-incorporated structured lipids with multiple regression and backward elimination. The coefficient of determination (R2) for the incorporation was 0.93, and that for the di-incorporated products was 0.94. The optimal conditions were flow rate, 2 ml/min; temperature, 65...

  2. Enzymatic interesterification on the physicochemical properties of Moringa oleifera seed oil blended with palm olein and virgin coconut oil

    Directory of Open Access Journals (Sweden)

    Dollah, S.

    2015-06-01

    Full Text Available The enzymatic interesterification (IE of palm olein (PO and virgin coconut oil (VCO with the high oleic acid (86% Moringa oleifera seed oil (MoO could yield a good source of oleic acid fat stock that may contain desirable nutritional and physical properties. Lipozyme RMIM resulted in different functionalities for the MoO/PO and MoO/VCO blends due to inherent changes in triacylglycerol (TAG compositions which, in turn, led to different trends in DSC thermograms and solid fat contents (SFC. The enzymatic IE of MoO/VCO increased U2S and S2U (up to 20% medium and long chain, MLCT while it decreased U3 (triunsaturated and S3 (trisaturated TAGs. The IE of the MoO/PO blends increased U2S and S3 (MMP, myristic, myristic, palmitic and decreased S2U, resulting in a lowering of melting points and SFC for MoO/VCO, while showing an increase in them for MoO/PO. A 2.55% increase in S3 after 24 h MoO/PO 30:70 IE revealed a 6.5% harder oil at 10 °C which may imply a wider application compared to the original liquid oils. Novel MLCTs with improved nutritional and physical properties were generated in the MoO/VCO blends after IE due to the incorporation of oleic acid and medium chain fatty acids. MoO/PO 50:50 and 70:30 w/w after 12 h IE and MoO/VCO 30:70 are suitable for incorporation into the fat phase in ice-cream formulations while, the spreadability and plasticity of MoO/VCO 70:30 improved at low temperatures. Both interesterified blends could be used as high oleic acid frying oils.La interesterificación enzimática (IE de la oleína de palma (PO, aceite de coco virgen (VCO con alto contenido en ácido oleico (86% y aceites de semilla de Moringa oleífera (MOO podría ser una buena fuente de ácido oleico con propiedades nutricionales y físicas deseables. La lipozyme RMIM produce diferentes funcionalidades para las mezclas MoO/PO y MoO/ VCO debido a los cambios inherentes en la composición de triacilgliceroles (TAG que, a su vez, dieron lugar a diferentes

  3. Nutritional enrichment of vegetable oils with long-chain n-3 fatty acids through enzymatic interesterification with a new vegetable lipase

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    Sousa, J. S.

    2015-06-01

    Full Text Available The aim of the present work was to produce vegetable oils enriched with long-chain n-3 fatty acids of nutraceutical interest, through an enzyme-catalyzed interesterification with a new lipase, from physic nut (Jatropha curcas L.. The Vegetable Lipase Powder (biocatalyst called VLP, which has never been applied in functional foods, was obtained from the physic nut seed, and efficiently hydrolyzed the 95% of waste fish oil in 24 h. Urea precipitation was used to concentrate polyunsaturated fatty acids (PUFA and was further interesterified with oils of different sources by means of enzymatic catalysis. After the interesterification reaction, which was also catalyzed by the VLP, the PUFA content in coconut oil increased almost ten-fold from 1.8% to 17.7%. In palm oil, the PUFA content increased two-fold from 10.5% to 21.8%, while in olive oil the level of PUFA increased from 8.6% to 21.3%. The mixture of eicosapentaenoic acid (EPA and docosahexaenoic acid (DHA (3.7% to 3.9% was incorporated into the triacylglycerol fraction of each of the coconut, palm and olive oils. Through the hydroesterification (hydrolysis followed by interesterification all the interesterified vegetable oils tested presented sufficient EPA and DHA levels to satisfy the levels recommended for intake by human adults in one tablespoon.El objetivo del presente trabajo fue producir aceites vegetales enriquecidos con ácidos grasos n-3 de cadena larga de interés nutraceutico, por interesterificación catalizada mediante una nueva lipasa, una enzima de semilla de Jatropha curcas L. La lipasa vegetal en polvo (biocatalizador llamada VLP, nunca ha sido aplicada en alimentos funcionales, se obtuvo mediante procedimientos físicos con semillas de nueces, e hidrolizó eficientemente el 95% de aceites de residuos de pescado en 24 h. La precipitación con urea se utilizó para concentrar los ácidos grasos poliinsaturados (PUFA que fueron posteriormente interesterificados con aceites de

  4. The properties of the mixture of beef tallow and rapeseed oil with a high content of tallow after chemical and enzymatic interesterification

    Directory of Open Access Journals (Sweden)

    Gruczynska, Eliza

    2005-12-01

    Full Text Available A mixture of beef tallow with rapeseed oil (3:1 wt/wt was interesterified using sodium metoxide or immobilized lipases from Rhizomucor miehei (Lipozyme IM and Candida antarctica (Novozym 435 as catalysts. Chemical interesterifications were carried out at 60 and 90 ºC for 0.5 and 1.5 h using 0.4, 0.6 and 1.0 wt-% CH3ONa. Depending on the catalyst used enzymatic interesterifications were carried out at 60 ºC for 8 h (Lipozyme IM or at 80 ºC for 4 h (Novozym 435. The catalysts doses were kept constant (8 % but the water content in catalysts varied from 2 to 10 %. The starting mixture and the interesterified products were separated by column chromatography into a pure triacylglycerol fraction and a non-triacylglycerol fraction, which contained free fatty acids, mono- and diacylglycerols. It was found that the concentrations of free fatty acids and partial acylglycerols increased after interesterification. The slip melting points and solid fat contents of the triacylglycerol fractions isolated from interesterified fats were lower when compared with nonesterified blends. The sn-2 and sn-1,3 distributions of fatty acids in the triacylglycerol fractions before and after interesterification were determined.These distributions were random after chemical interesterification and near random when Novozym 435 was used. When Lipozyme IM was used, the fatty acid composition at the sn-2 position remained practically unchanged compared with the starting blend. The interesterified fats and isolated triacylglycerols had reduced oxidative stability, as assessed by Rancimat induction times. The addition of 0.02 % of BHA or BHT to the interesterified fats improved their stabilitie.Una mezcla de sebo con aceite de colza (3:1 p/p fue interesterificada usando metóxido de sodio y lipasas inmovilizadas de Rhizomucor miehei (Lipozyme IM and Candida antarctica (Novozym 435 como catalizadores. La interesterificación química se llevó a cabo a 60 ºC y 90

  5. Human milk fat substitute from butterfat: production by enzymatic interesterification and evaluation of oxidative stability

    DEFF Research Database (Denmark)

    Sørensen, Ann-Dorit Moltke; Xu, Xuebing; Zhang, Long

    2010-01-01

    Recent data have suggested that the fatty acid composition and molecular structure of fats in infant formulas should be as similar to human milk fat as possible to obtain optimal fat and calcium absorption from the infant formula. This work investigated the possibilities of using enzyme technology...... and butterfat as a material to produce a fat similar to human milk fat with respect to the above parameters. Moreover, the oxidative stability of the enzyme modified human milk fat substitute (HMFS) was compared to the fat blend used for the production of HMFS. Using a combination of enzyme technology......, fractionation and batch deodorization and with butterfat in combination with soybean oil and rapeseed oil as raw materials it was possible to produce HMFS with a molecular structure and fatty acid composition that was very similar to that of human milk fat. The oxidative stability of the HMFS oil was lower than...

  6. Effects of Degree of Enzymatic Interesterification on the Physical Properties of Margarine Fats: Solid Fat Content, Crystallization Behavior, Crystal Morphology, and Crystal Network

    DEFF Research Database (Denmark)

    Zhang, Hong; Smith, Paul; Adler-Nissen, Jens

    2004-01-01

    In this study enzymatic-interesterified margarine fats with different conversion degrees were produced in a packed-bed reactor. The effects of conversion degree on the formation of free fatty acids and diacyglycerols, solid fat content, crystallization behavior, microstructure, and crystal network...... °C with increasing conversion degree. Increased conversion degree from the blend to products, measured by X-ray with addition of 50% of rapeseed oil for dilution, caused the content of â to decrease from 100% to 33%, and 30% and eventually to pure ⢠crystal. However, double chain packing...

  7. Chemical interesterification of soybean oil and fully hydrogenated soybean oil: Influence of the reaction time

    International Nuclear Information System (INIS)

    Ribeiro, Ana Paula Badan; Masuchi, Monise Helen; Grimaldi, Renato; Goncalves, Lireny Aparecida Guaraldo

    2009-01-01

    Chemical interesterification is an important alternative to produce zero trans fats. In practice, however, excessive reaction times are used to ensure complete randomization. This work evaluated the influence of the reaction time on the interesterification of soybean oil/fully hydrogenated soybean oil blend, carried out in the following conditions: 100 deg C, 500 rpm stirring speed, 0.4% (w/w) sodium methoxide catalyst. The triacylglycerol composition, solid fat content and melting point analysis showed that the reaction was very fast, reaching the equilibrium within 5 min. This result suggests the interesterification can be performed in substantially lower times, with reduction in process costs. (author)

  8. Monitoring lipase-catalyzed interesterification for bulky fats modification with FT-IR/NIR spectroscopy

    DEFF Research Database (Denmark)

    Chang, Tinghong; Lai, Xuxin; Zhang, Hong

    2005-01-01

    This work demonstrates the application of FT-IR and FT-NIR spectroscopy to monitor the enzymatic interesterification process for bulky fat modification. The reaction was conducted between palm stearin and coconut oil (70/30, w/w) with the catalysis of Lipozyme TL IM at 70°C in a batch reactor...

  9. Monitoring lipase-catalyzed butterfat interesterification with rapesee oil by Fourier transform near-infrared spectroscopy

    DEFF Research Database (Denmark)

    Zhang, Hong; Mu, Huiling; Xu, Xuebing

    2006-01-01

    This work demonstrates the application of FT-NIR spectroscopy to monitor the enzymatic interesterification process for butterfat modification. The reactions were catalyzed by Lipozyme TL IM at 70 C for the blend of butterfat/rapeseed oil (70/30, w/w) in a packed-bed reactor. The blend and intere...

  10. Improved enzymatic production of phenolated glycerides through alkyl phenolate intermediate

    DEFF Research Database (Denmark)

    Yang, Zhiyong; Feddern, Vivian; Glasius, Marianne

    2011-01-01

    This work reported a novel approach for synthesis of dihydrocaffoylated glycerides, consisting of 2 steps: enzymatic synthesis of octyl dihydrocaffeate (as a synthetic intermediate) from octanol and dihydrocaffeic acid (DHCA), and enzymatic interesterification of triglycerides with octyl dihydroc......This work reported a novel approach for synthesis of dihydrocaffoylated glycerides, consisting of 2 steps: enzymatic synthesis of octyl dihydrocaffeate (as a synthetic intermediate) from octanol and dihydrocaffeic acid (DHCA), and enzymatic interesterification of triglycerides with octyl...

  11. Enzymatically interesterified fats based on mutton tallow and walnut oil suitable for cosmetic emulsions.

    Science.gov (United States)

    Kowalska, M; Mendrycka, M; Zbikowska, A; Stawarz, S

    2015-02-01

    Formation of emulsion systems based on interesterified fats was the objective of the study. Enzymatic interesterification was carried out between enzymatic mutton tallow and walnut oil in the proportions 2 : 3 (w/w) to produce fats not available in nature. At the beginning of the interesterification process, the balance between the interesterification and fat hydrolysis was intentionally disturbed by adding more water to the catalyst (Lipozyme IR MR) of the reaction to produce more of the polar fraction monoacylglycerols [MAGs] and diacylglycerols [DAGs]. To obtain a greater quantity of MAGs and DAGs in the reaction environment via hydrolysis, water was added (11, 13, 14, 16 w-%) to the enzymatic preparation. The obtained fats were used to form emulsions. The emulsions were evaluated with respect to sensory and skin moisturizing properties by 83 respondents. Determination of emulsion stability using temperature and centrifugal tests was carried out. Morphology and the type of emulsions were determined. The respondents described the skin to which the emulsions in testing were applied as smooth, pleasant to touch and adequately moisturized. The work has demonstrated that interesterification of a mutton tallow and walnut oil blend resulted in new fats with very interesting characteristics of triacylglycerols that are not present in the environment. The results of the present work indicate the possibility of application of fats with the largest quantity of MAGs and DAGs as a fat base of emulsions in the cosmetic industries. The hypothesis assumed in this work of producing additional quantities of MAGs and DAGs (in the process of enzymatic interesterification) responsible for the stability of the system was confirmed. It should be pointed out that the emulsions based on interesterified fats exhibited a greater level of moisturization of the skin than the emulsions containing non-interesterified fat. Also, in the respondents' opinion, the emulsion containing fat, which

  12. Covalent immobilization of lipase onto aminopropyl-functionalized hydroxyapatite-encapsulated-γ-Fe2O3 nanoparticles: A magnetic biocatalyst for interesterification of soybean oil.

    Science.gov (United States)

    Xie, Wenlei; Zang, Xuezhen

    2017-07-15

    Hydroxyapatite-encapsulated γ-Fe 2 O 3 nanoparticles were prepared, and lipase from Candida rugosa was then covalently bound onto the magnetic materials via covalent linkages. The magnetic carrier and immobilized lipase were characterized by enzyme activity assays, XRD, FT-IR, TEM, VSM and N 2 adsorption-desorption techniques. Results demonstrated that γ-Fe 2 O 3 nanoparticles were coated with the hydroxyapatite, and the lipase was indeed tethered to the magnetic carriers without damage to their structure. The immobilized lipase showed a strong magnetic responsiveness and displayed high catalytic activities towards the interesterification of soybean oil. The interesterified products were evaluated for their total fatty acid (FA) composition, slip melting point (SMP), iodine value, triacylglycerols (TAGs) profile and FA composition at sn-2 position in TAGs. The FA positional distributions and TAG species significantly changed after the enzymatic interesterification. Besides this, the interesterified products showed an obvious reduction in their SMP in comparison with the physical blends. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. Triglyceride selectivity of immobilized Thermomyces lanuginosa lipase in interesterification

    DEFF Research Database (Denmark)

    Rønne, Torben Harald; Pedersen, Lars S.; Xu, Xuebing

    2005-01-01

    from tri-C4:0 to tri-C20:0, except for tri-C6:0, and in a series of unsaturated FA from tri-C18:1 to tri-C18:3. The quantification was performed by HPLC, and different methods of selectivity evaluation were used. None of the methods used showed any significant differences between the performances......The triglyceride (fatty acid) selectivity of an immobilized lipase from Thermomyces lanuginosa (Lipozyme TL IM) was investigated in lipase-catalyzed interesterification reactions between two mono-acid TG in n-hexane. Tristearin (tri-C18:0) was used as a reference in a series of TG with saturated FA...

  14. Clinical evaluation of chemokine and enzymatic biomarkers of Gaucher disease

    NARCIS (Netherlands)

    Deegan, Patrick B.; Moran, Mary Teresa; McFarlane, Ian; Schofield, J. Paul; Boot, Rolf G.; Aerts, Johannes M. F. G.; Cox, Timothy M.

    2005-01-01

    Purpose: Gaucher disease is an exemplary orphan disorder. Enzyme replacement therapy with imiglucerase is effective, but very expensive. To improve the assessment of severity of disease and responses to this costly treatment, we have evaluated several enzymatic biomarkers and a newly-described

  15. Process Evaluation Tools for Enzymatic Cascades Welcome Message

    DEFF Research Database (Denmark)

    Abu, Rohana

    improvement and implementation. Hence, the goal of this thesis is to evaluate the process concepts in enzymatic cascades in a systematic manner, using tools such as thermodynamic and kinetic analysis. Three relevant case studies have been used to exemplify the approach. In the first case study, thermodynamic......Biocatalysis is attracting significant attention from both academic and industrial scientists due to the excellent capability of enzyme to catalyse selective reactions. Recently, much interest has been shown in the application of enzymatic cascades as a useful tool in organic synthesis......, the kinetics can be controlled in a highly efficient way to achieve a sufficiently favourable conversion to a given target product. This is exemplified in the second case study, in the kinetic modelling of the formation of 2-ketoglutarate from glucoronate, the second case study. This cascade consists of 4...

  16. Interesterification of rapeseed oil catalyzed by tin octoate

    International Nuclear Information System (INIS)

    Galia, Alessandro; Centineo, Alessio; Saracco, Guido; Schiavo, Benedetto; Scialdone, Onofrio

    2014-01-01

    The interesterification of rapeseed oil was performed for the first time by using tin octoate as Lewis acid homogeneous catalysts and methyl or ethyl acetate as acyl acceptors in a batch reactor, within the temperature range 393–483 K. The yields in fatty acid ethyl esters (FAEE) and triacetin (TA) after 20 h of reaction time increased from 8% and 2%–to 61% and 22%, respectively, when the reaction temperature increased from 423 to 483 K. An optimum value of 40 for the acyl acceptor to oil molar ratio was found to be necessary to match good fatty acid alkyl ester yields with high enough reaction rate. The rate of generation of esters was significantly higher when methyl acetate was used as acyl acceptor instead of its ethyl homologue. The collected results suggest that tin octoate can be used as effective catalyst for the interesterification of rapeseed oil with methyl or ethyl acetate being highly soluble in the reaction system, less expensive than enzymes and allowing the operator to work under milder conditions than supercritical interesterification processes. - Highlights: • We study the interesterification of rapeseed oil catalyzed by tin(II) octoate. • Tin(II) octoate is an effective homogeneous catalyst at 483 K. • The acyl acceptor to oil molar ratio must be optimized. • Higher rate of reaction is obtained with methyl acetate as acyl acceptor

  17. Multicenter evaluation of an enzymatic method for glycated albumin.

    Science.gov (United States)

    Paleari, Renata; Bonetti, Graziella; Callà, Cinzia; Carta, Mariarosa; Ceriotti, Ferruccio; Di Gaetano, Nicola; Ferri, Marilisa; Guerra, Elena; Lavalle, Gabriella; Cascio, Claudia Lo; Martino, Francesca Gabriela; Montagnana, Martina; Moretti, Marco; Santini, Gabriele; Scribano, Donata; Testa, Roberto; Vero, Anna; Mosca, Andrea

    2017-06-01

    The use of glycated albumin (GA) has been proposed as an additional glycemic control marker particularly useful in intermediate-term monitoring and in situation when HbA 1c test is not reliable. We have performed the first multicenter evaluation of the analytical performance of the enzymatic method quantILab Glycated Albumin assay implemented on the most widely used clinical chemistry analyzers (i.e. Abbott Architect C8000, Beckman Coulter AU 480 and 680, Roche Cobas C6000, Siemens ADVIA 2400 and 2400 XPT). The repeatability of the GA measurement (expressed as CV, %) implemented in the participating centers ranged between 0.9% and 1.2%. The within-laboratory CVs ranged between 1.2% and 1.6%. A good alignment between laboratories was found, with correlation coefficients from 0.996 to 0.998. Linearity was confirmed in the range from 7.6 to 84.7%. The new enzymatic method for glycated albumin evaluated by our investigation is suitable for clinical use. Copyright © 2017 Elsevier B.V. All rights reserved.

  18. Radiochemical methods for studying lipase-catalyzed interesterification of lipids

    International Nuclear Information System (INIS)

    Schuch, R.; Mukherjee, K.D.

    1987-01-01

    Reactions involving lipase-catalyzed interesterification of lipids, which are of commendable interest in biotechnology, have been monitored and assayed by radiochemical methods using 14 C-labeled substrates. Medium chain (C 12 plus C 14 ) triacylglycerols were reacted in the presence of an immobilized lipase from Mucor miehei and hexane at 45 0 C with methyl [1- 14 C]oleate, [1- 14 C]oleic acid, [carboxyl- 14 C]trioleoylglycerol, [1- 14 C]octadecenyl alcohol, and [U- 14 C]glycerol, each of known specific activity. The reactions were monitored and the rate of interesterification determined by radio thin layer chromatography from the incorporation of radioactivity into acyl moieties of triacylglycerols (from methyl oleate, oleic acid, and trioleoylglycerol), alkyl moieties of wax esters (from octadecenyl alcohol), and into glycerol backbone of monoacylglycerols and diacylglycerols (from glycerol). (orig.)

  19. Enzymatic modification of egg lecithin to improve properties.

    Science.gov (United States)

    Asomaning, Justice; Curtis, Jonathan M

    2017-04-01

    This research studied the enzymatic modification of egg yolk phospholipids and its effect on physicochemical properties. Egg yolk lipids were extracted with food grade ethanol and egg phospholipids (ePL) produced by deoiling with acetone. Vegetable oils were used to interesterify ePL utilizing Lipozyme®: sn-1,3 specific lipase. The enzymatic interesterification resulted in a single phase liquid product, whereas simple blending of the ePL and vegetable oil resulted in a product with two phases. In addition solid fat content decreased by 50% at -10°C and 94% at 35°C when compared with egg yolk lipids extract. A decrease in melting temperature resulted from the interesterification process. Interesterification improved emulsion stability index when used as an emulsifier in oil-in-water emulsion and compared to the native and soy lecithin. Enzyme reusability test showed retention of 63% activity after 10 cycles. Overall, the properties of native egg phospholipids were significantly enhanced in a potentially useful manner through interesterification. Copyright © 2016 Elsevier Ltd. All rights reserved.

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

    DEFF Research Database (Denmark)

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

    2004-01-01

    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...

  1. Evaluation of enzymatic reactors for large-scale panose production.

    Science.gov (United States)

    Fernandes, Fabiano A N; Rodrigues, Sueli

    2007-07-01

    Panose is a trisaccharide constituted by a maltose molecule bonded to a glucose molecule by an alpha-1,6-glycosidic bond. This trisaccharide has potential to be used in the food industry as a noncariogenic sweetener, as the oral flora does not ferment it. Panose can also be considered prebiotic for stimulating the growth of benefic microorganisms, such as lactobacillus and bifidobacteria, and for inhibiting the growth of undesired microorganisms such as E. coli and Salmonella. In this paper, the production of panose by enzymatic synthesis in a batch and a fed-batch reactor was optimized using a mathematical model developed to simulate the process. Results show that optimum production is obtained in a fed-batch process with an optimum production of 11.23 g/l h of panose, which is 51.5% higher than production with batch reactor.

  2. The effect of interesterification on the bioavailability of fatty acids in structured lipids.

    Science.gov (United States)

    Farfán, M; Villalón, M J; Ortíz, M E; Nieto, S; Bouchon, P

    2013-08-15

    Fatty acid (FA) profile is a critical factor in the nutritional properties of fats, but, stereochemistry may also play a fundamental role in the rate and extent to which FAs are absorbed and become available. To better understand this phenomenon, we evaluated the bioavailability of FAs in linseed-oil and palm-stearin blends compared to their interesterified mix, using a sn-1,3 stereospecific lipase, to determine if there was any difference in terms of FA availability when using this technology. Test meals were fed through an intragastric feeding tube on Sprague-Dawley male rats after 18 h fasting. Postprandial blood samples were collected after meal or physiological serum (control) administration and the FA profile of plasma lipids was determined. Results showed that modification of the melting profile through interesterification, without altering the bioavailability determined by sn-2 stereochemistry, could delay lipid absorption at the beginning, but had no effect on total lipid absorption. Copyright © 2013. Published by Elsevier Ltd.

  3. Production of specific structured lipids by enzymatic interesterification: optimization of the reaction by response surface design

    DEFF Research Database (Denmark)

    Xu, Xuebing; Skands, Anja Rebecca Havegaard; Adler-Nissen, Jens

    1998-01-01

    Rapeseed oil and capric acid were interesterified in solvent-free media catalyzed by Lipozyme IM (Rhizomucor miehei) to produce specific-structured lipids (SSLs). The process was optimized by response surface design concerning the effects of acyl migration and the by-products of diacylglycerols...

  4. Pilot scale intensification of rubber seed (Hevea brasiliensis) oil via chemical interesterification using hydrodynamic cavitation technology.

    Science.gov (United States)

    Bokhari, Awais; Yusup, Suzana; Chuah, Lai Fatt; Klemeš, Jiří Jaromír; Asif, Saira; Ali, Basit; Akbar, Majid Majeed; Kamil, Ruzaimah Nik M

    2017-10-01

    Chemical interesterification of rubber seed oil has been investigated for four different designed orifice devices in a pilot scale hydrodynamic cavitation (HC) system. Upstream pressure within 1-3.5bar induced cavities to intensify the process. An optimal orifice plate geometry was considered as plate with 1mm dia hole having 21 holes at 3bar inlet pressure. The optimisation results of interesterification were revealed by response surface methodology; methyl acetate to oil molar ratio of 14:1, catalyst amount of 0.75wt.% and reaction time of 20min at 50°C. HC is compared to mechanical stirring (MS) at optimised values. The reaction rate constant and the frequency factor of HC were 3.4-fold shorter and 3.2-fold higher than MS. The interesterified product was characterised by following EN 14214 and ASTM D 6751 international standards. Copyright © 2017 Elsevier Ltd. All rights reserved.

  5. Synthesis of structured triacylglycerols containing medium-chain and long-chain fatty acids by interesterification with a stereoespecific lipase from Mucor miehei.

    Directory of Open Access Journals (Sweden)

    Nieto, Susana

    1999-06-01

    Full Text Available The preparation of structured triacylglycerols sn-1, sn-3 dilauryl, sn-2 eicosapentaenoyl glycerol and sn-1, sn-3 dilauryl, sn-2 docosahexaenoyl glycerol by enzymatic interesterification under restricted water availability is described. Laurie acid, one of the substrates for interesterification, was obtained by the controlled hydrolysis of coconut oil by a non-specific lipase obtained from Candida cylindracea. The fatty acid was separated from the hydrolysis products by silverresin column chromatography and converted to methyl ester, sn-2 Eicosapentaenoyl glycerol and sn-2 docosahexaenoyl glycerol were prepared by the hydrolysis of fish oil by the sn-1, sn-3 stereospecific immobilized lipase Lipozyme IM-20 obtained from Mucor miehei as described in the accompanying paper. The interesterification was carried out in a water jacketed glass reactor and the triacylglycerol products were separated and recovered through aluminum oxide column chromatography The interesterification procedure described allows to obtain In laboratory scale structured triacylglycerols containing medium-chain fatty acids at the sn-1 and sn-3 positions and long-chain polyunsaturated fatty acid from marine origin at the sn-2 glycerol position.

    Se describe la preparación de triacilgliceroles estructurados sn-1, sn-3 dilauril, sn-2 ecosapentaenoil glicerol y sn-1, sn-3 diiauril, sn-2 docosahexaenoil glicerol por interesterificación enzimática bajo disponibilidad de agua reducida. Acido láurico, uno de los sustratos para la interesterificación, se obtuvo mediante hidrólisis controlada del aceite de coco por una lipasa no-específica obtenida de Candida cylindracea. Los ácidos grasos se separaron de los productos de hidrólisis mediante cromatografía en columna de resina de plata y convertidos en sus esteres metílicos, sn-2 Eicosapentaenoil glicerol y sn-2 docosahexaenoil glicerol se prepararon mediante hidrólisis de aceite de pescado por la sn-1, sn

  6. Enzymatic production of alkyl esters through alcoholysis: A critical evaluation of lipases and alcohols

    DEFF Research Database (Denmark)

    Li, Deng; Xu, Xuebing; Gudmundur G, Haraldsson

    2005-01-01

    This paper focuses on a detailed evaluation of commercially available immobilized lipases and simple monohydric alcohols for the production of alkyl esters from sunflower oil by enzymatic alcoholysis. Six lipases were tested with seven alcohols, including straight and branched-chain primary...... in an increased degree of conversion for all lipases except Novozym 435. The secondary alcohol 2-propanol significantly reduced the alcoholysis reaction with all lipases; however, the branch-chain isobutanol was more advantageous than linear 1-butanol for Novozym 435, Lipozyme RM IM, and Lipase PS-C. Many...

  7. Evaluation of physical structural features on influencing enzymatic hydrolysis efficiency of micronized wood

    Science.gov (United States)

    Jinxue Jiang; Jinwu Wang; Xiao Zhang; Michael Wolcott

    2016-01-01

    Enzymatic hydrolysis of lignocellulosic biomass is highly dependent on the changes in structural features after pretreatment. Mechanical milling pretreatment is an effective approach to alter the physical structure of biomass and thus improve enzymatic hydrolysis. This study examined the influence of structural characteristics on the enzymatic hydrolysis of micronized...

  8. Process evaluation of enzymatic hydrolysis with filtrate recycle for the production of high concentration sugars.

    Science.gov (United States)

    Xue, Ying; Rusli, Jannov; Chang, Hou-Min; Phillips, Richard; Jameel, Hasan

    2012-02-01

    Process simulation and lab trials were carried out to demonstrate and confirm the efficiency of the concept that recycling hydrolysate at low total solid enzymatic hydrolysis is one of the options to increase the sugar concentration without mixing problems. Higher sugar concentration can reduce the capital cost for fermentation and distillation because of smaller retention volume. Meanwhile, operation cost will also decrease for less operating volume and less energy required for distillation. With the computer simulation, time and efforts can be saved to achieve the steady state of recycling process, which is the scenario for industrial production. This paper, to the best of our knowledge, is the first paper discussing steady-state saccharification with recycling of the filtrate form enzymatic hydrolysis to increase sugar concentration. Recycled enzymes in the filtrate (15-30% of the original enzyme loading) resulted in 5-10% higher carbohydrate conversion compared to the case in which recycled enzymes were denatured. The recycled hydrolysate yielded 10% higher carbohydrate conversion compared to pure sugar simulated hydrolysate at the same enzyme loading, which indicated hydrolysis by-products could boost enzymatic hydrolysis. The high sugar concentration (pure sugar simulated) showed inhibition effect, since about 15% decrease in carbohydrate conversion was observed compared with the case with no sugar added. The overall effect of hydrolysate recycling at WinGEMS simulated steady-state conditions with 5% total solids was increasing the sugar concentration from 35 to 141 g/l, while the carbohydrate conversion was 2% higher for recycling at steady state (87%) compared with no recycling strategy (85%). Ten percent and 15% total solid processes were also evaluated in this study.

  9. An integrated model for the technical and economic evaluation of an enzymatic biomass conversion process

    Energy Technology Data Exchange (ETDEWEB)

    Nguyen, Q A; Saddler, J N [Forintek Canada Corp., Ottawa, Ontario (CA). Dept. of Biotechnology and Chemistry

    1991-01-01

    A process simulation model was constructed, using the Lotus 123 spreadsheet, to evaluate the technical and economic feasibility of a lignocellulosic-to-ethanol bioconversion process. The major components of the process were: steam pretreatment, fractionation, enzyme production, enzymatic hydrolysis, ethanol fermentation of glucose and xylose, product recovery, and waste treatment. The model provided an estimation of the price of ethanol and evaluated the interdependence of process parameters and their effect on the production cost of ethanol. A case study of a plant processing 500 tonnes of aspenwood per day showed that the parameters which had the most significant impact on the price of ethanol were: the cost of wood, cost of enzymes, efficiency of cellulose hydrolysis, ethanol yield from xylose, efficiency of the fractionation process, and the selling price of the lignin by-product. (author).

  10. Evaluation of lime and hydrothermal pretreatments for efficient enzymatic hydrolysis of raw sugarcane bagasse.

    Science.gov (United States)

    Grimaldi, Maira Prearo; Marques, Marina Paganini; Laluce, Cecília; Cilli, Eduardo Maffud; Sponchiado, Sandra Regina Pombeiro

    2015-01-01

    Ethanol production from sugarcane bagasse requires a pretreatment step to disrupt the cellulose-hemicellulose-lignin complex and to increase biomass digestibility, thus allowing the obtaining of high yields of fermentable sugars for the subsequent fermentation. Hydrothermal and lime pretreatments have emerged as effective methods in preparing the lignocellulosic biomass for bioconversion. These pretreatments are advantageous because they can be performed under mild temperature and pressure conditions, resulting in less sugar degradation compared with other pretreatments, and also are cost-effective and environmentally sustainable. In this study, we evaluated the effect of these pretreatments on the efficiency of enzymatic hydrolysis of raw sugarcane bagasse obtained directly from mill without prior screening. In addition, we evaluated the structure and composition modifications of this bagasse after lime and hydrothermal pretreatments. The highest cellulose hydrolysis rate (70 % digestion) was obtained for raw sugarcane bagasse pretreated with lime [0.1 g Ca(OH)2/g raw] for 60 min at 120 °C compared with hydrothermally pretreated bagasse (21 % digestion) under the same time and temperature conditions. Chemical composition analyses showed that the lime pretreatment of bagasse promoted high solubilization of lignin (30 %) and hemicellulose (5 %) accompanied by a cellulose accumulation (11 %). Analysis of pretreated bagasse structure revealed that lime pretreatment caused considerable damage to the bagasse fibers, including rupture of the cell wall, exposing the cellulose-rich areas to enzymatic action. We showed that lime pretreatment is effective in improving enzymatic digestibility of raw sugarcane bagasse, even at low lime loading and over a short pretreatment period. It was also demonstrated that this pretreatment caused alterations in the structure and composition of raw bagasse, which had a pronounced effect on the enzymes accessibility to the

  11. Environmentally evaluated HPLC-ELSD method to monitor enzymatic synthesis of a non-ionic surfactant.

    Science.gov (United States)

    Gaber, Yasser; Akerman, Cecilia Orellana; Hatti-Kaul, Rajni

    2014-01-01

    N-Lauroyl-N-methylglucamide is a biodegradable surfactant derived from renewable resources. In an earlier study, we presented an enzymatic solvent-free method for synthesis of this compound. In the present report, the HPLC method developed to follow the reaction between lauric acid/methyl laurate and N-methyl glucamine (MEG) and its environmental assessment are described. Use of ultraviolet (UV) absorption or refractive index (RI) detectors did not allow the detection of N-methyl glucamine (MEG). With Evaporative light scattering detector ELSD, it was possible to apply a gradient elution, and detect MEG with a limit of detection, LOD = 0.12 μg. A good separation of the peaks: MEG, lauric acid, product (amide) and by-product (amide-ester) was achieved with the gradient program with a run time of 40 min. The setting of ELSD detector was optimized using methyl laurate as the analyte. LC-MS/MS was used to confirm the amide and amide-ester peaks. We evaluated the greenness of the developed method using the freely available software HPLC-Environmental Assessment Tool (HPLC-EAT) and the method got a scoring of 73 HPLC-EAT units, implying that the analytical procedure was more environmentally benign compared to some other methods reported in literature whose HPLC-EAT values scored up to 182. Use of ELSD detector allowed the detection and quantification of the substrates and the reaction products of enzymatic synthesis of the surfactant, N-lauroyl-N-methylglucamide. The developed HPLC method has acceptable environmental profile based on HPLC-EAT evaluation.

  12. Evaluation of peroxidases from roots of Cyperus hermaphroditus as enzymatic mechanisms in phenanthrene oxidation

    Energy Technology Data Exchange (ETDEWEB)

    Guerrero Zuniga, A. [Inst. Mexicano del Petroleo, Mexico City (Mexico). Environmental Protection Management Office; Rodriguez Dorantes, A.M. [Lab. Fisiologia Vegetal, Escuela Nacional de Ciencias Biologicas, Mexico City (Mexico). Depto Botanica

    2006-07-01

    Although phenanthrene is not mutagenic or carcinogenic, it has been shown to be toxic to aquatic organisms. This study evaluated in-vitro phenanthrene oxidation by peroxidases from radical extracts of Cyperus hermaphroditus plants. The characterization of oxidation products of phenanthrene related to the induction of root peroxidases was also examined. Concentrated ethanol stock of phenanthrene solution was added to the mineral solution of each plant container. The total radical biomass was placed in 4.5 ml of an ionic solution to analyze the enzymatic activity of the extracellular peroxidases. The total protein for each experiment was quantified by the Bradford method. Extracellular peroxidases activity was measured using the spectrophotometric method. The amount of radical biomass was quantified as high in the 80 and 120 ppm phenanthrene treatments relative to the control plants. It was suggested that the nature of the Cyperaceae roots combined with the high-octanol water coefficient and a low water solubility for phenanthrene may have facilitated the stabilization of the contaminant towards the roots. The ability of Cyperus hermaphroditus to immobilize phenanthrene through its adhesion was encouraged by the conditions of the hydroponic culture system. The adsorption of phenanthrene was increased with the time of exposure to the contaminant due to the greater total root mass. The study also showed the transformation of phenanthrene by radical extracts of Cyperus hermaphroditus containing guaiacol peroxidases with 12 per cent residual phenanthrene in the in vitro assays. The spectrophotometric analysis confirmed that the enzymatic systems are responsible for the phytotransformation of the pollutant. 9 refs., 2 tabs., 5 figs.

  13. Lipozyme IM-catalyzed interesterification for the production of margarine fats in a 1 kg scale stirred tank reactor

    DEFF Research Database (Denmark)

    Zhang, Hong; Xu, Xuebing; Mu, Huiling

    2000-01-01

    Lipozyme IM-catalyzed interesterification of the oil blend between palm stearin and coconut oil (75/25 w/w) was studied for the production of margarine fats in a 1 kg scale batch stirred tank reactor. Parameters such as lipase load, water content, temperature, and reaction time were investigated...

  14. Parameters affecting diacylglycerol formation during the production of specific-structured lipids by lipase-catalyzed interesterification

    DEFF Research Database (Denmark)

    Xu, Xuebing; Mu, Huiling; Skands, Anja

    1999-01-01

    Diacylglycerols (DAGs) are important intermediates in lipase-catalyzed interesterification, but a high DAG concentration in the reaction mixture results in a high DAG content in the final product. We have previously shown that a high DAG concentration in the reaction mixture increases the degree ...

  15. Interesterification reaction activity, fatty acid composition and selectivity ratio of soybean oil

    Directory of Open Access Journals (Sweden)

    El-Shattory, Y.

    1998-12-01

    Full Text Available The interesterification reaction was carried out by adding oleic acid to soybean oil by ratio 1:2 w/w under different conditions of temperature, stirring and catalyst percentages. Assessment of the interesterification of oils was reported by determination of saponification value, iodine value and fatty acids composition. This study showed that linolenic acid which is responsible for flavour instability of soybean oil and consider as primary factor contributing to deterioration of this oil could be reduced to less than or equals 3%.

    Se han llevado a cabo reacciones de interesterificación mediante la adición de ácido oleico a aceite de soja en la relación 1:2 w/w bajo diferentes condiciones de temperatura, agitación y porcentaje de catalizador. La evaluación de la interesterificación de los aceites se realizó por determinación del índice de saponificación, el índice de iodo y la composición en ácidos grasos. Este estudio mostró que el ácido linolénico, que es responsable de la inestabilidad del flavor del aceite de soja y considerado como factor primario que contribuye a la deterioración de este aceite, podría ser reducido a cantidades menores o iguales al 3%.

  16. Cloning, expression, and enzymatic activity evaluation of cholesterol oxidase gene isolated from a native Rhodococcus sp.

    Directory of Open Access Journals (Sweden)

    Hamed Esmaeil Lashgarian

    2016-10-01

    Full Text Available Cholesterol oxidase (CHO is one of the valuable enzymes that play an important role in: measurement of serum cholesterol, food industry as a biocatalyst and agriculture as a biological larvicide. This enzyme was produced by several bacterial strains. Wild type enzyme produced by Rhodococcus sp. secret two forms of CHO enzyme: extra cellular and membrane bound type which its amount is low and unstable. The goal of the study was cloning, expression, and enzymatic activity evaluation of cholesterol oxidase gene isolated from a native Rhodococcus sp. CHO gene was isolated from native bacteria and cloned into pET23a. In the next step, the construct was expressed in E.coli BL21 and induced by different concentration of IPTG ranges from 0.1 - 0.9 mM. This gene contains 1642 bp and encodes a protein consists of 533 amino acids. It has about 96 % homology with CHO gene isolated from Rhodococcus equi. The high expression was obtained in 0.5 mM concentration of IPTG after 4 hour induction. This recombinant enzyme had a molecular weight of 55 kDa, that secretion of intra cellular type is much more than extracellular form. The optimum pH and temperature conditions for the recombinant enzyme were 7.5 and 45°C, respectively. CHO enzyme obtained from Rhodococcus sp. is a cheap enzyme with medical and industrial applications that can be produced easily and purified in large scale with simple methods.

  17. Effect of micella interesterification on fatty acids composition and volatile components of soybean and rapeseed oils

    Directory of Open Access Journals (Sweden)

    Afifi, Sherine M.

    2000-10-01

    Full Text Available Micella interesterification of soybean and rapeseed oils was carried out using 0.2, 0.4 and 0.6 percentages of nickel catalyst, each at different temperatures of 60, 90 and 120ºC for 2, 4, and 6 hours. The proposed interesterification reaction conditions to obtain an oil with low linoleic acid level were 0.2 % nickel catalyst at 120ºC for 4 hours, 0.4% nickel catalyst at 90ºC for 4 hours and 0.6% at 60ºC for 4 hours. Fatty acid composition and chemical analysis of the interesterified and non-esterified oils were estimated. Selected samples undergo heating at 180ºC for 4 hours determining the volatile components. The appearance of some components supported the interesterification process for modification of fatty acid constituents of the oils.Se ha llevado a cabo la interesterificación en fase miscelar de aceites de soja y de colza usando un 0.2%, 0.4% y 0.6% de níquel como catalizador, a diferentes temperaturas (60, 90 y 120ºC durante 2, 4 y 6 horas. Las condiciones de reacción de interesterificación propuestas para obtener un aceite con niveles de ácidos linolénicos bajos fueron 0.2 % de níquel a 120ºC durante 4 horas, 0.4 % de níquel a 90ºC durante 4 horas y 0.6 % a 60ºC durante 4 horas. Se han estimado la composición en ácidos grasos y el análisis químico de los aceites interesterificados y no-esterificados. Las muestras seleccionadas se sometieron a calentamiento a 180ºC durante 4 horas determinando los componentes volátiles. La aparición de algunos componentes apoyó el proceso de interesterificación por modificación de los ácidos grasos constituyentes de los aceites.

  18. Storage stability of margarines produced from enzymatically interesterified fats compared to those prepared by conventional methods - Chemical properties

    DEFF Research Database (Denmark)

    Zhang, Hong; Jacobsen, Charlotte; Pedersen, Lars Saaby

    2006-01-01

    margarines in a pilot plant. Storage stability studies were carried out at storage temperatures of 5 and 25øC for 12wk. Margarines from the enzymatically interesterified fats were compared to the margarines produced by the conventional methods (chemical interesterification and physical blending......In this study, four margarine hardstocks were produced, two from enzymatically interesterified fats at 80 and 100% conversion, one from chemically randomized fat and one from physically mixed fat. These four hardstocks, blended with 50% sunflower oil, were mainly used for the production of table...... interesterified fat had higher PV in weeks4, 8 and10 than the margarines produced from the enzymatically interesterified fats and the physically blended fat. These differences were not caused by different contents of tocopherols in the hardstocks. The differences between the processes for chemical and enzymatic...

  19. Production of structured lipid with a low omega-6/omega-3 fatty acids ratio by enzymatic interesterification

    International Nuclear Information System (INIS)

    Ilyasoglu, H.

    2017-01-01

    A structured lipid (SL) constituting omega fatty acids was synthesized by using linseed and grape seed oils as substrates via a lipase-catalyzed reaction. Lipozyme® TL IM was used as a biocatalyst. Good quadratic models predicting the incorporation of omega fatty acids were achieved via the Response surface methodology (RSM). The optimal conditions for targeted omega-6/omega-3 fatty acid ratio (2:1) were obtained at a substrate molar ratio 1.4, time 8.4 h, and enzyme amount 6.4%. The SL contained linoleic acid (43 g 100g-1), which was mainly located in the sn-2 position (40 g 100g-1). α-Linoleic acid, and α-linolenic acid at the sn-2 position were 22 g 100g-1, and 11 g 100g-1, respectively. The oxidative stability of the SL, and SL with antioxidants was also investigated. The produced SL may be proposed as a source of a balanced intake of omega fatty acids and an ingredient in functional food formulations. [es

  20. A Miniature Membrane Reactor for Evaluation of Process Design Options on the Enzymatic Degradation of Pectin

    DEFF Research Database (Denmark)

    Zainal Alam, Muhd Nazrul Hisham; Pinelo, Manuel; Arnous, Anis

    2011-01-01

    was fabricated from poly(methylmethacrylate) (PMMA) and poly(dimethylsiloxane) (PDMS) with a working volume of ∼190 μL. The prototype also contained the necessary sensors and actuators, i.e., pressure transducer, mixing via magnetic stirrer bar and a temperature controller. The functionality of the prototype...... was demonstrated by performing a continuous enzymatic degradation of pectin experiment for a range of reactor conditions: different membrane molecular weight cutoff (MWCO) values, enzyme-to-substrate ratios (E/S), and substrate feeding rates (F) were assessed. Based on the experimental data, it was found...

  1. Evaluation of soluble fraction and enzymatic residual fraction of dilute dry acid, ethylenediamine, and steam explosion pretreated corn stover on the enzymatic hydrolysis of cellulose.

    Science.gov (United States)

    Qin, Lei; Liu, Li; Li, Wen-Chao; Zhu, Jia-Qing; Li, Bing-Zhi; Yuan, Ying-Jin

    2016-06-01

    This study is aimed to examine the inhibition of soluble fraction (SF) and enzymatic residual fraction (ERF) in dry dilute acid (DDA), ethylenediamine (EDA) and steam explosion (SE) pretreated corn stover (CS) on the enzymatic digestibility of cellulose. SF of DDA, EDA and SE pretreated CS has high xylose, soluble lignin and xylo-oligomer content, respectively. SF of EDA pretreated CS leads to the highest inhibition, followed by SE and DDA pretreated CS. Inhibition of ERF of DDA and SE pretreated CS is higher than that of EDA pretreated CS. The inhibition degree (A0/A) of SF is 1.76 and 1.21 times to that of ERF for EDA and SE pretreated CS, respectively. The inhibition degree of ERF is 1.05 times to that of SF in DDA pretreated CS. The quantitative analysis shows that SF of EDA pretreated CS, SF and ERF of SE pretreated CS cause significant inhibition during enzymatic hydrolysis. Copyright © 2016 Elsevier Ltd. All rights reserved.

  2. Hydrothermal treatment and enzymatic hydrolysis of Tamarix ramosissima: evaluation of the process as a conversion method in a biorefinery concept.

    Science.gov (United States)

    Xiao, Ling-Ping; Shi, Zheng-Jun; Xu, Feng; Sun, Run-Cang

    2013-05-01

    The present work investigated the effects of hydrothermal treatment (HTT) of Tamarix ramosissima by determination of sugar and inhibitor formation in the liquid fraction, and chemical and morphological changes of the pretreated solid material coupled with an evaluation of enzymatic hydrolysis. HTT was carried out in a batch reactor system at a maximal temperature (TMAX 180-240 °C) and evaluated for severities logRo ranging from 2.40 to 4.17. The liquid fractions were analyzed by HPLC, GPC, and GC-MS. The morphology and composition of the solid residues were characterized using an array of techniques, such as SEM, XRD, BET surface area, and CP/MAS (13)C NMR. Using a variety of tools, we have developed a better understanding of how HTT process affects biomass structure and cellulose properties that impact on its digestibility. These results provided new insights into the factors limiting enzymatic digestibility and mechanism of biomass deconstruction during hydrothermal process. Crown Copyright © 2012. Published by Elsevier Ltd. All rights reserved.

  3. Evaluation of oil removal efficiency and enzymatic activity in some fungal strains for bioremediation of petroleum-polluted soils

    Directory of Open Access Journals (Sweden)

    Mohsenzadeh Fariba

    2012-12-01

    Full Text Available Abstract Background Petroleum pollution is a global disaster and there are several soil cleaning methods including bioremediation. Methods In a field study, fugal strains were isolated from oil-contaminated sites of Arak refinery (Iran and their growth ability was checked in potato dextrose agar (PDA media containing 0-10% v/v crude oil, the activity of three enzymes (Catalase, Peroxidase and Phenol Oxidase was evaluated in the fungal colonies and bioremediation ability of the fungi was checked in the experimental pots containing 3 kg sterilized soil and different concentrations of petroleum (0-10% w/w. Results Four fungal strains, Acromonium sp., Alternaria sp., Aspergillus terreus and Penicillium sp., were selected as the most resistant ones. They were able to growth in the subjected concentrations and Alternaria sp. showed the highest growth ability in the petroleum containing media. The enzyme assay showed that the enzymatic activity was increased in the oil-contaminated media. Bioremediation results showed that the studied fungi were able to decrease petroleum pollution. The highest petroleum removing efficiency of Aspergillus terreus, Penicillium sp., Alternaria sp. and Acromonium sp. was evaluated in the 10%, 8%, 8% and 2% petroleum pollution respectively. Conclusions Fungi are important microorganisms in decreasing of petroleum pollution. They have bioremediation potency that is related to their enzymatic activities.

  4. Evaluation of Oil Removal Efficiency and Enzymatic Activity in Some fungal Strains for Bioremediation of Petroleum-Polluted Soils

    Directory of Open Access Journals (Sweden)

    Fariba Mohsenzadeh

    2012-12-01

    Full Text Available Background: Petroleum pollution is a global disaster and there are several soil cleaning methods including bioremediation.Methods: In a field study, fugal strains were isolated from oil-contaminated sites of Arak refinery (Iran and their growth ability was checked in potato dextrose agar (PDA media containing 0-10% v/v crude oil, the activity of three enzymes (Catalase, Peroxidase and Phenol Oxidase was evaluated in the fungal colonies and bioremediation ability of the fungi was checked in the experimental pots containing 3 kg sterilized soil and different concentrations of petroleum (0-10% w/w.Results: Four fungal strains, Acromonium sp., Alternaria sp., Aspergillus terreus and Penicillium sp., were selected asthe most resistant ones. They were able to growth in the subjected concentrations and Alternaria sp. showed thehighest growth ability in the petroleum containing media. The enzyme assay showed that the enzymatic activity was increased in the oil-contaminated media. Bioremediation results showed that the studied fungi were able to decrease petroleum pollution. The highest petroleum removing efficiency of Aspergillus terreus, Penicillium sp.,Alternaria sp. and Acromonium sp. was evaluated in the 10%, 8%, 8% and 2% petroleum pollution respectively.Conclusions: Fungi are important microorganisms in decreasing of petroleum pollution. They have bioremediation potency that is related to their enzymatic activities.

  5. Evaluation of Enzymatically Modified Soy Protein Isolate Film Forming Solution and Film at Different Manufacturing Conditions.

    Science.gov (United States)

    Mohammad Zadeh, Elham; O'Keefe, Sean F; Kim, Young-Teck; Cho, Jin-Hun

    2018-04-01

    The effects of transglutaminase on soy protein isolate (SPI) film forming solution and films were investigated by rheological behavior and physicochemical properties based on different manufacturing conditions (enzyme treatments, enzyme incubation times, and protein denaturation temperatures). Enzymatic crosslinking reaction and changes in molecular weight distribution were confirmed by viscosity measurement and SDS-PAGE, respectively, compared to 2 controls: the nonenzyme treated and the deactivated enzyme treated. Films treated with both the enzyme and the deactivated enzyme showed significant increase in tensile strength (TS), percent elongation (%E), and initial contact angle of films compared to the nonenzyme control film due to the bulk stabilizers in the commercial enzyme. Water absorption property, protein solubility, Fourier transform infrared (FTIR) and X-ray diffraction (XRD) spectroscopy revealed that enzyme treated SPI film matrix in the molecular structure level, resulted in the changes in physicochemical properties. Based on our observation, the enzymatic treatment at appropriate conditions is a practical and feasible way to control the physical properties of protein based biopolymeric film for many different scientific and industrial areas. Enzymes can make bridges selectively among different amino acids in the structure of protein matrix. Therefore, protein network is changed after enzyme treatment. The behavior of biopolymeric materials is dependent on the network structure to be suitable in different applications such as bioplastics applied in food and pharmaceutical products. In the current research, transglutaminase, as an enzyme, applied in soy protein matrix in different types of forms, activated and deactivated, and different preparation conditions to investigate its effects on different properties of the new bioplastic film. © 2018 Institute of Food Technologists®.

  6. Evaluation of alcohol dehydrogenase and aldehyde dehydrogenase enzymes as bi-enzymatic anodes in a membraneless ethanol microfluidic fuel cell

    Science.gov (United States)

    Galindo-de-la-Rosa, J.; Arjona, N.; Arriaga, L. G.; Ledesma-García, J.; Guerra-Balcázar, M.

    2015-12-01

    Alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (AldH) enzymes were immobilized by covalent binding and used as the anode in a bi-enzymatic membraneless ethanol hybrid microfluidic fuel cell. The purpose of using both enzymes was to optimize the ethanol electro-oxidation reaction (EOR) by using ADH toward its direct oxidation and AldH for the oxidation of aldehydes as by-products of the EOR. For this reason, three enzymatic bioanode configurations were evaluated according with the location of enzymes: combined, vertical and horizontally separated. In the combined configuration, a current density of 16.3 mA cm-2, a voltage of 1.14 V and a power density of 7.02 mW cm-2 were obtained. When enzymes were separately placed in a horizontal and vertical position the ocp drops to 0.94 V and to 0.68 V, respectively. The current density also falls to values of 13.63 and 5.05 mA cm-2. The decrease of cell performance of bioanodes with separated enzymes compared with the combined bioanode was of 31.7% and 86.87% for the horizontal and the vertical array.

  7. Determination of photosynthetic and enzymatic biomarkers sensitivity used to evaluate toxic effects of copper and fludioxonil in alga Scenedesmus obliquus

    International Nuclear Information System (INIS)

    Dewez, David; Geoffroy, Laure; Vernet, Guy; Popovic, Radovan

    2005-01-01

    Modulated PAM fluorometry and Plant Efficiency Analyser methods were used to investigate photosynthetic fluorescence parameters of alga Scenedesmus obliquus exposed to inhibitory effect of fungicides copper sulphate and fludioxonil (N-(4-nitrophenyl)-N'-propyl-uree). The change of those parameters were studied when alga S. obliquus have been exposed during 48 h to different concentrations of fungicides (1, 2 and 3 mg l -1 ). Under the same condition, enzymatic activities of catalase, ascorbate peroxidase, glutathione reductase and glutathione S-transferase were investigated to evaluate antioxidative response to fungicides effects. The change of sensitivity of those parameters was dependent to the mode of fungicide action, their concentration and time of exposure. For copper effects, the most indicative photosynthetic biomarkers were parameters Q N as non-photochemical fluorescence quenching, Q Emax as the proton induced fluorescence quenching and ABS/RC as the antenna size per photosystem II reaction center. Copper induced oxidative stress was indicated by increased activity of catalase serving as the most sensitive and valuable enzymatic biomarker. On the other hand, fludioxonil effect on photosynthetic parameters was very negligible and consequently not very useful as biomarkers. However, fludioxonil induced strong antioxidative activities associated with cytosol enzymes, as we found for catalase, ascorbate peroxidase and glutathione S-transferase activities. By obtained results, we may suggest for the activation of those enzymes to be sensitive and valuable biomarkers of oxidative stress induced by fludioxonil. Determination of biomarkers sensitivity may offer advantages in providing real criteria to use them for ecotoxicological diagnostic studies

  8. Pilot batch production of specific-structured lipids by lipase-catalyzed interesterification: preliminary study on incorporation and acyl migration

    DEFF Research Database (Denmark)

    Xu, Xuebing; Balchen, Steen; Høy, Carl-Erik

    1998-01-01

    Effects of water content, reaction time and their relationships in the production of two types of specific-structured lipids (sn-MLM- and sn-LML-types: L-long chain fatty acids; M-medium chain fatty acids) by lipase-catalyzed interesterification in a solvent-free system were studied...... of two totally position-opposed lipids can be observed. Presumably these are caused by the different chain length of the fatty acids. The relationships between reaction time and water content are inverse and give a quantitative prediction of incorporation and acyl migration in selected reaction...

  9. Empirical evaluation of inhibitory product, substrate, and enzyme effects during the enzymatic saccharification of lignocellulosic biomass.

    Science.gov (United States)

    Smith, Benjamin T; Knutsen, Jeffrey S; Davis, Robert H

    2010-05-01

    The cellulose hydrolysis kinetics during batch enzymatic saccharification are typified by a rapid initial rate that subsequently decays, resulting in incomplete conversion. Previous studies suggest that changes associated with the solution, substrate, or enzymes may be responsible. In this work, kinetic experiments were conducted to determine the relative magnitude of these effects. Pretreated corn stover (PCS) was used as a lignocellulosic substrate likely to be found in a commercial saccharification process, while Avicel and Kraft lignin were used to create model substrates. Glucose inhibition was observed by spiking the reaction slurry with glucose during initial-rate experiments. Increasing the glucose concentration from 7 to 48 g/L reduced the cellulose conversion rate by 94%. When product sugars were removed using ultrafiltration with a 10 kDa membrane, the glucose-based conversion increased by 9.5%. Reductions in substrate reactivity with conversion were compared directly by saccharifying PCS and Avicel substrates that had been pre-reacted to different conversions. Reaction of substrate with a pre-conversion of 40% resulted in about 40% reduction in the initial rate of saccharification, relative to fresh substrate with identical cellulose concentration. Overall, glucose inhibition and reduced substrate reactivity appear to be dominant factors, whereas minimal reductions of enzyme activity were observed.

  10. Oxyfluorfen toxic effect on S. obliquus evaluated by different photosynthetic and enzymatic biomarkers.

    Science.gov (United States)

    Geoffroy, L; Dewez, D; Vernet, G; Popovic, R

    2003-11-01

    The effect of oxyfluorfen was investigated when alga Scenedesmus obliquus has been exposed to different concentrations (7.5, 15, and 22.5 microg x L(-1)) at 12, 24, and 48 hours of exposure. Toxicity test was done by using 13 biomarkers concerning growth rate, chlorophyll content and indicators of photosynthetic and antioxidant enzyme activities. The change of the 13 parameters showed a great variation of sensitivity indicating differences in parameters' suitability to be used as biomarkers when alga culture was exposed to oxyfluorfen toxicity. The order of sensitivity between those biomarkers was: Antenna size (ABS/RC) > Chlorophyll content > Catalase (CAT) > Operational PSII quantum yield (phiS(PSII)) > Glutathione S-transferase (GST) > Functional plastoquinone pool (Q(PQ)) > Glutathione reductase (GR) > Growth rate > Nonphotochemical quenching (QN) > Proton gradient quenching (Q(Emax)) > Ascorbate peroxidase (APX) > Photochemical quenching (Q(p)) > Maximum PSII quantum yield (Phi(PSII)). The effect of oxyfluorfen on the changes of those parameters was interpreted as a result of herbicide mode of action at molecular level of alga cellular system. This study indicated for some photosynthetic and enzymatic biomarkers to be useful indicators of toxicity effect induced in non-target alga species. Determination of biomarkers' sensitivity order may facilitate their selection to be used in environmental risk assessment of polluted water.

  11. Evaluation of a third party enzymatic ammonia method for use on the Roche Cobas 6000 (c501) automated platform.

    Science.gov (United States)

    Seiden-Long, Isolde; Schnabl, Kareena; Skoropadyk, Wendy; Lennon, Nola; McKeage, Arlayne

    2014-08-01

    Adaptation of the Randox Enzymatic Manual UV Ammonia method to be used on the Roche Cobas 6000 (c501) automated analyzer platform. The Randox ammonia reagent was evaluated for precision, linearity, accuracy and interference from hemolysis, icterus and lipemia on the Roche c501 analyzer. Comparison studies were conducted for the Randox reagent between Roche c501, Siemens Vista, Ortho Vitros 250, and Beckman DxC methods. The Randox reagent demonstrates acceptable within-run (L1=65 μmol/L, CV 3.4% L2=168 μmol/L, CV 1.9%) and between-run precision (L1=29 μmol/L, CV 7.3% L2=102 μmol/L, CV 3.0%), Analytical Measurement Range (7-940 μmol/L), and accuracy. The method interference profile is superior for the Randox method (hemolysis index up to 600, icteric index up to 60, lipemic index up to 100) as compared to the Roche method (hemolysis index up to 200, icteric index up to 10, lipemic index up to 50). Comparison was very good between the Randox reagent and two other wet chemistry platforms. The Randox Enzymatic Manual UV Ammonia reagent is an available alternative to the Roche Cobas c501 reagent. The method is more robust to endogenous interferences and less prone to instrument error flags, thus allowing the majority of clinical specimens to be reported without additional sample handling at our institution. Copyright © 2014 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

  12. Determination of photosynthetic and enzymatic biomarkers sensitivity used to evaluate toxic effects of copper and fludioxonil in alga Scenedesmus obliquus

    Energy Technology Data Exchange (ETDEWEB)

    Dewez, David [Departement de Chimie et de Biochimie, Centre TOXEN, Universite du Quebec a Montreal, CP 8888, Succursale Centre-Ville, Montreal, Quebec, H3C 3P8 (Canada); Geoffroy, Laure [Laboratoire d' Eco-Toxicologie, Unite de recherche ' Vignes et Vins de Champagne' , UPRES-EA 2069, Universite de Reims Champagne-Ardenne BP 1039, F51687 REIMS CEDEX 2 (France); Vernet, Guy [Laboratoire d' Eco-Toxicologie, Unite de recherche ' Vignes et Vins de Champagne' , UPRES-EA 2069, Universite de Reims Champagne-Ardenne BP 1039, F51687 REIMS CEDEX 2 (France); Popovic, Radovan [Departement de Chimie et de Biochimie, Centre TOXEN, Universite du Quebec a Montreal, CP 8888, Succursale Centre-Ville, Montreal, Quebec, H3C 3P8 (Canada)]. E-mail: popovic.radovan@uqam.ca

    2005-08-30

    Modulated PAM fluorometry and Plant Efficiency Analyser methods were used to investigate photosynthetic fluorescence parameters of alga Scenedesmus obliquus exposed to inhibitory effect of fungicides copper sulphate and fludioxonil (N-(4-nitrophenyl)-N'-propyl-uree). The change of those parameters were studied when alga S. obliquus have been exposed during 48 h to different concentrations of fungicides (1, 2 and 3 mg l{sup -1}). Under the same condition, enzymatic activities of catalase, ascorbate peroxidase, glutathione reductase and glutathione S-transferase were investigated to evaluate antioxidative response to fungicides effects. The change of sensitivity of those parameters was dependent to the mode of fungicide action, their concentration and time of exposure. For copper effects, the most indicative photosynthetic biomarkers were parameters Q {sub N} as non-photochemical fluorescence quenching, Q {sub Emax} as the proton induced fluorescence quenching and ABS/RC as the antenna size per photosystem II reaction center. Copper induced oxidative stress was indicated by increased activity of catalase serving as the most sensitive and valuable enzymatic biomarker. On the other hand, fludioxonil effect on photosynthetic parameters was very negligible and consequently not very useful as biomarkers. However, fludioxonil induced strong antioxidative activities associated with cytosol enzymes, as we found for catalase, ascorbate peroxidase and glutathione S-transferase activities. By obtained results, we may suggest for the activation of those enzymes to be sensitive and valuable biomarkers of oxidative stress induced by fludioxonil. Determination of biomarkers sensitivity may offer advantages in providing real criteria to use them for ecotoxicological diagnostic studies.

  13. Otimização da reação de interesterificação química do óleo de palma Optimization of the cemical interesterification reaction of palm oil

    OpenAIRE

    Renato Grimaldi; Lireny Aparecida Guaraldo Gonçalves; Marlene Yumi Ando

    2005-01-01

    The Brazilian market has been showing a growing concern with nutritional values of oil components of foods. Chemical interesterification is a promising alternative to the current processes of modifying the consistency of oils. Chemical interesterification of deodorized palm oil was studied on a laboratory scale. The best results were obtained with 0.4% MeONa and heating for 20 min at 100 °C. These conditions are based on the largest variation in triacylglycerols as compared to a control. ...

  14. Evaluation of single and stack membraneless enzymatic fuel cells based on ethanol in simulated body fluids.

    Science.gov (United States)

    Galindo-de-la-Rosa, J; Arjona, N; Moreno-Zuria, A; Ortiz-Ortega, E; Guerra-Balcázar, M; Ledesma-García, J; Arriaga, L G

    2017-06-15

    The purpose of this work is to evaluate single and double-cell membraneless microfluidic fuel cells (MMFCs) that operate in the presence of simulated body fluids SBF, human serum and blood enriched with ethanol as fuels. The study was performed using the alcohol dehydrogenase enzyme immobilised by covalent binding through an array composed of carbon Toray paper as support and a layer of poly(methylene blue)/tetrabutylammonium bromide/Nafion and glutaraldehyde (3D bioanode electrode). The single MMFC was tested in a hybrid microfluidic fuel cell using Pt/C as the cathode. A cell voltage of 1.035V and power density of 3.154mWcm -2 were observed, which is the highest performance reported to date. The stability and durability were tested through chronoamperometry and polarisation/performance curves obtained at different days, which demonstrated a slow decrease in the power density on day 10 (14%) and day 20 (26%). Additionally, the cell was tested for ethanol oxidation in simulated body fluid (SBF) with ionic composition similar to human blood plasma. Those tests resulted in 0.93V of cell voltage and a power density close to 1.237mWcm -2 . The double cell MMFC (Stack) was tested using serum and human blood enriched with ethanol. The stack operated with blood in a serial connection showed an excellent cell performance (0.716mWcm -2 ), demonstrating the feasibility of employing human blood as energy source. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. STUDY OF THE PREPARATION OF SUGAR FROM HIGH-LIGNIN LIGNOCELLULOSE APPLYING SUBCRITICAL WATER AND ENZYMATIC HYDROLYSIS: SYNTHESIS AND CONSUMABLE COST EVALUATION

    Directory of Open Access Journals (Sweden)

    HANNY F. SANGIAN

    2015-05-01

    Full Text Available This study concern sugars hydrolyzed from the high-lignin coconut coir dust using moderate subcritical water (SCW hydrolysis at pressures 20-40 bar for 1 h and to evaluate the consumable costs driver generated. The SCW method produced two products, sugar liquid and solid (SCW-treated substrate. The solid was proceeded to prepare the sugar via enzymatic hydrolysis using pure cellulase. Yield of sugar hydrolyzed from lignocellulose by SCW technique was 0.25 gram sugar/gram cellulose +hemicellulose, or 0.09-gram sugar/gram lignocellulose at 160 °C and 40 bar. While, the maximum yield of sugar liberated enzymatically from SCW-treated solid was 0.35-gram sugar/gram cellulose+hemicellulose, or 0.13-gram sugar/gram SCW-treated solid. It was found that carbon dioxide gas was the highest cost driving in SCW hydrolysis.

  16. [Evaluations of newborn screening program performance and enzymatic diagnosis of glucose-6-phosphate dehydrogenase deficiency in Guangzhou].

    Science.gov (United States)

    Tang, F; Huang, Y L; Jiang, X; Jia, X F; Li, B; Feng, Y; Chen, Q Y; Tang, C F

    2018-05-02

    Objective: To reveal the molecular epidemiologic characteristics of glucose-6-phosphate dehydrogenase (G6PD) gene and to evaluate based on the genetic analysis the newborn screening program performance and enzymatic diagnosis of G6PD deficiency in Guangzhou. Methods: G6PD enzyme activities were measured by quantitative fluorescence assay in dry blood spots of 16 319 newborns(8 725 males, 7 594 females) 3-7 days after birth in Guangzhou Newborn Center. They were born in Guangzhou form Oct. 1 to 20, 2016. The cutoff value of G6PD was less than 2.6 U/g Hb in dry blood spots. G6PD deficiency was diagnosed when G6PDblood cells. Genetic analysis of G6PD gene was performed on the dry blood spot samples of 823 newborns (including positive 346, negative 477)with various levels of G6PD enzyme activities through fluorescence PCR melting curve analysis(FMCA) to detect 15 kinds of mutations reported to be common among Chinese.G6PD gene Sanger sequency was performed in seven highly suspicious patients with negative results by FMCA. Results: (1) Using the cutoff value of G6PDT, c.551C>T, c.835A>T hemizygote were found in 3 male's samples, respectively. (3) The estimated prevalence of harboring mutation was 6.0% in males and 13.5% in females according to rates of mutation in samples with various levels of G6PD enzyme activities. Six common mutations were c.1388G>A、c.1376G>T, c.95A> G, c.871G>A, c.1024C>T, c.392G>T, accounting for 95.5% of detected alleles .(4) based on results of G6PD gene analysis, the newborn scereening of G6PD deficiency with cutoff value G6PDblood cells were 95.5%, 97.2%, respectively. Conclusions: The prevalence of G6PD deficiency in males was 6.0% in Guangzhou. Six mutations c.1388G>A, c.1376G>T, c.95A>G, c.871G>A, c.1024C>T, c.392G>T accounted for 95.5%. The cutoff value of G6PD<2.6 U/g Hb innewborn screening program and the criteria of biochemical diagnosis could accurately identify G6PD deficiency . Combined with biochemical and molecular analysis will

  17. Evaluation of a next generation direct whole blood enzymatic assay for hemoglobin A1c on the ARCHITECT c8000 chemistry system.

    Science.gov (United States)

    Teodoro-Morrison, Tracy; Janssen, Marcel J W; Mols, Jasper; Hendrickx, Ben H E; Velmans, Mathieu H; Lotz, Johannes; Lackner, Karl; Lennartz, Lieselotte; Armbruster, David; Maine, Gregory; Yip, Paul M

    2015-01-01

    The utility of HbA1c for the diagnosis of type 2 diabetes requires an accurate, precise and robust test measurement system. Currently, immunoassay and HPLC are the most popular methods for HbA1c quantification, noting however the limitations associated with some platforms, such as imprecision or interference from common hemoglobin variants. Abbott Diagnostics has introduced a fully automated direct enzymatic method for the quantification of HbA1c from whole blood on the ARCHITECT chemistry system. Here we completed a method evaluation of the ARCHITECT HbA1c enzymatic assay for imprecision, accuracy, method comparison, interference from hemoglobin variants and specimen stability. This was completed at three independent clinical laboratories in North America and Europe. The total imprecision ranged from 0.5% to 2.2% CV with low and high level control materials. Around the diagnostic cut-off of 48 mmol/mol, the total imprecision was 0.6% CV. Mean bias using reference samples from IFCC and CAP ranged from -1.1 to 1.0 mmol/mol. The enzymatic assay also showed excellent agreement with HPLC methods, with slopes of 1.01 and correlation coefficients ranging from 0.984 to 0.996 compared to Menarini Adams HA-8160, Bio-Rad Variant II and Variant II Turbo instruments. Finally, no significant effect was observed for erythrocyte sedimentation or interference from common hemoglobin variants in patient samples containing heterozygous HbS, HbC, HbD, HbE, and up to 10% HbF. The ARCHITECT enzymatic assay for HbA1c is a robust and fully automated method that meets the performance requirements to support the diagnosis of type 2 diabetes.

  18. Immobilization of Lipase using Alginate Hydrogel Beads and Enzymatic Evaluation in Hydrolysis of p-Nitrophenol Butyrate

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Shuang; Shang, Wenting; Yang, Xiaoxi; Zhang, Shujuan; Zhang, Xiaogang; Chen, Jiawei [Renmin Univ. of China, Beijing (China)

    2013-09-15

    The immobilization of enzyme is one of the key issues both in the field of enzymatic research and industrialization. In this work, we reported a facile method to immobilize Candida Antarctica lipase B (CALB) in alginate carrier. In the presence of calcium cation, the enzyme-alginate suspension could be cross-linked to form beads with porous structure at room temperature, and the enzyme CALB was dispersed in the beads. Activity of the enzyme-alginate composite was verified by enzymatic hydrolysis reaction of p-nitrophenol butyrate in aqueous phase. The effects of reaction parameters such as temperature, pH, embedding and lyophilized time on the reactive behavior were discussed. Reuse cycle experiments for the hydrolysis of p-nitrophenol butyrate demonstrated that activity of the enzyme-alginate composite was maintained without marked deactivation up to 6 repeated cycles.

  19. Evaluation of jumbo squid (Dosidicus gigas byproduct hydrolysates obtained by acid-enzymatic hydrolysis and by autohydrolysis in practical diets for Pacific white shrimp (Litopenaeus vannamei

    Directory of Open Access Journals (Sweden)

    Mayra Lizett González-Félix

    2014-09-01

    Full Text Available The marine bioprocessing industry offers great potential to utilize byproducts for fish meal replacement in aquafeeds. Jumbo squid is an important fishery commodity in Mexico, but only the mantle is marketed. Head, fins, guts and tentacles are discarded in spite of being protein-rich byproducts. This study evaluated the use of two jumbo squid byproduct hydrolysates obtained by acid-enzymatic hydrolysis (AEH and by autohydrolysis (AH as ingredients in practical diets for shrimp. The hydrolysates were included at levels of 2.5 and 5.0% of the diet dry weight in four practical diets, including a control diet without hydrolysate. Shrimp growth and survival were not significantly affected by the dietary treatments. Postharvest quality of abdominal muscle was evaluated in terms of proximate composition and sensory evaluation. Significantly higher crude protein was observed in the muscle of shrimp fed the highest hydrolysate levels, AH 5% (204.8 g kg- 1 or AEH 5% (201.3 g kg- 1. Sensory analysis of cooked muscle showed significant differences for all variables evaluated: color, odor, flavor, and firmness. It was concluded that Jumbo squid byproducts can be successfully processed by autohydrolysis or acid-enzymatic hydrolysis, and that up to 5.0% of the hydrolysates can be incorporated into shrimp diets without affecting growth or survival.

  20. Evaluation of relationships between growth rate, tree size, lignocellulose composition and enzymatic saccharification in interspecific Corymbia hybrids and parental taxa.

    Directory of Open Access Journals (Sweden)

    Adam L Healey

    2016-11-01

    Full Text Available In order for a lignocellulosic bioenergy feedstock to be considered sustainable, it must possess a high rate of growth to supply biomass for conversion. Despite the desirability of a fast growth rate for industrial application, it is unclear what effect growth rate has on biomass composition or saccharification. We characterized Klason lignin, glucan, and xylan content with response to growth in Corymbia interspecific F1 hybrid families (HF and parental species C. torelliana (CT and C. citriodora subspecies variegata (CCV and measured the effects on enzymatic hydrolysis from hydrothermally pretreated biomass. Analysis of biomass composition within Corymbia populations found similar amounts of Klason lignin content (19.7-21.3% among parental and hybrid populations, whereas glucan content was clearly distinguished within CCV (52% and HF148 (60% as compared to other populations (28-38%. Multiple linear regression indicates that biomass composition is significantly impacted by tree size measured at the same age, with Klason lignin content increasing with diameter breast height (DBH (+0.12% per cm DBH increase, and glucan and xylan typically decreasing per DBH cm increase (-0.7% and -0.3%, respectively. Polysaccharide content within CCV and HF-148 were not significantly affected by tree size. High-throughput enzymatic saccharification of hydrothermally pretreated biomass found significant differences among Corymbia populations for total glucose production from biomass, with parental CT and hybrids HF-148 and HF-51 generating the highest amounts of glucose (~180 mg/g biomass, respectively, with HF-51 undergoing the most efficient glucan-to-glucose conversion (74%. Based on growth rate, biomass composition, and further optimization of enzymatic saccharification yield, high production Corymbia hybrid trees are potentially suitable for fast-rotation bioenergy or biomaterial production.

  1. Evaluation of Relationships between Growth Rate, Tree Size, Lignocellulose Composition, and Enzymatic Saccharification in Interspecific Corymbia Hybrids and Parental Taxa.

    Science.gov (United States)

    Healey, Adam L; Lee, David J; Lupoi, Jason S; Papa, Gabriella; Guenther, Joel M; Corno, Luca; Adani, Fabrizio; Singh, Seema; Simmons, Blake A; Henry, Robert J

    2016-01-01

    In order for a lignocellulosic bioenergy feedstock to be considered sustainable, it must possess a high rate of growth to supply biomass for conversion. Despite the desirability of a fast growth rate for industrial application, it is unclear what effect growth rate has on biomass composition or saccharification. We characterized Klason lignin, glucan, and xylan content with response to growth in Corymbia interspecific F1 hybrid families (HF) and parental species Corymbia torelliana and C. citriodora subspecies variegata and measured the effects on enzymatic hydrolysis from hydrothermally pretreated biomass. Analysis of biomass composition within Corymbia populations found similar amounts of Klason lignin content (19.7-21.3%) among parental and hybrid populations, whereas glucan content was clearly distinguished within C. citriodora subspecies variegata (52%) and HF148 (60%) as compared to other populations (28-38%). Multiple linear regression indicates that biomass composition is significantly impacted by tree size measured at the same age, with Klason lignin content increasing with diameter breast height (DBH) (+0.12% per cm DBH increase), and glucan and xylan typically decreasing per DBH cm increase (-0.7 and -0.3%, respectively). Polysaccharide content within C. citriodora subspecies variegata and HF-148 were not significantly affected by tree size. High-throughput enzymatic saccharification of hydrothermally pretreated biomass found significant differences among Corymbia populations for total glucose production from biomass, with parental Corymbia torelliana and hybrids HF-148 and HF-51 generating the highest amounts of glucose (~180 mg/g biomass, respectively), with HF-51 undergoing the most efficient glucan-to-glucose conversion (74%). Based on growth rate, biomass composition, and further optimization of enzymatic saccharification yield, high production Corymbia hybrid trees are potentially suitable for fast-rotation bioenergy or biomaterial production.

  2. Enzymatic production of wheat and ryegrass derived xylooligosaccharides and evaluation of their in vitro effect on pig gut microbiota

    DEFF Research Database (Denmark)

    Dotsenko, Gleb; Meyer, Anne S.; Canibe, Nuria

    2017-01-01

    This study examines enzymatic production of linear xylooligosaccharides (XOS) and branched arabinoxylooligosaccharides (AXOS) from monocotyledonous biomass, wheat straw and ryegrass, and compares the in vitro effects of these XOS and AXOS on pig gut microbiota. XOS and AXOS were obtained from...... the biomass by treatment with different endo-1,4-β-xylanases. XOS of DP2-6 from wheat straw, obtained after treatment with Aspergillus niger endo GH11, suppressed growth of Clostridium perfringens and resulted in a high level of lactic acid production when fermented in vitro by pig fecal microbiota...... (GH11). These results indicate that wheat straw as well as green grass biomass such as ryegrass have potential as new sources of putative prebiotics for pig feed....

  3. Biodegradation study of enzymatically catalyzed interpenetrating polymer network: Evaluation of agrochemical release and impact on soil fertility

    Directory of Open Access Journals (Sweden)

    Saruchi

    2016-03-01

    Full Text Available A novel interpenetrating polymer network (IPN has been synthesized through enzymatic initiation using lipase as initiator, glutaraldehyde as cross-linker, acrylic acid as primary monomer and acrylamide as secondary monomer. Biodegradability of synthesized interpenetrating polymer network was studied through soil burial and composting methods. Synthesized hydrogel was completely degraded within 70 days using composting method, while it was 86.03% degraded within 77 days using soil burial method. This was confirmed by Fourier transform Infrared spectroscopy (FTIR and Scanning electron microscopy (SEM techniques. Synthesized interpenetrating polymer network hydrogel was used as a device for controlled release of urea and also act as water releasing device. Their impact on soil fertility and plant growth was also studied. The initial diffusion coefficient has a greater value than the later diffusion coefficient indicating a higher fertilizer release rate during the early stage. Fertilizer release kinetic was also studied which showed Non-Fickian diffusion behavior, as the rate of fertilizer release was comparable to the relaxation time of the synthesized matrix. Synthesized IPN enhance the water uptake capacity up to 6.2% and 7.2% in sandy loam and clay soil, respectively.

  4. Response surface methodology for evaluation and optimization of process parameter and antioxidant capacity of rice flour modified by enzymatic extrusion.

    Science.gov (United States)

    Xu, Enbo; Pan, Xiaowei; Wu, Zhengzong; Long, Jie; Li, Jingpeng; Xu, Xueming; Jin, Zhengyu; Jiao, Aiquan

    2016-12-01

    For the purpose of investigating the effect of enzyme concentration (EC), barrel temperature (BT), moisture content (MC), and screw speed (SS) on processing parameters (product temperature, die pressure and special mechanical energy (SME)) and product responses (extent of gelatinization (GE), retention rate of total phenolic content (TPC-RR)), rice flour extruded with thermostable α-amylase was analyzed by response surface methodology. Stepwise regression models were computed to generate response surface and contour plots, revealing that both TPC-RR and GE increased as increasing MC while expressed different sensitivities to BT during enzymatic extrusion. Phenolics preservation was benefited from low SME. According to multiple-factor optimization, the conditions required to obtain the target SME (10kJ/kg), GE (100%) and TPC-RR (85%) were: EC=1.37‰, BT=93.01°C, MC=44.30%, and SS=171.66rpm, with the actual values (9.49kJ/kg, 99.96% and 87.10%, respectively) showing a good fit to the predicted values. Crown Copyright © 2016. Published by Elsevier Ltd. All rights reserved.

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

  6. Enzymatic modification of starch

    DEFF Research Database (Denmark)

    Jensen, Susanne Langgård

    In the food industry approaches for using bioengineering are investigated as alternatives to conventional chemical and physical starch modification techniques in development of starches with specific properties. Enzyme-assisted post-harvest modification is an interesting approach to this, since...... it is considered a clean and energy saving technology. This thesis aimed to investigate the effect of using reaction conditions, simulating an industrial process, for enzymatic treatment of starch with branching enzyme (BE) from Rhodothermus obamensis. Thus treatements were conducted at 70°C using very high...... substrate concentration (30-40% dry matter (DM)) and high enzyme activity (750-2250 BE units (BEU)/g sample). Starches from various botanical sources, representing a broad range of properties, were used as substrates. The effects of the used conditions on the BE-reaction were evaluated by characterization...

  7. A dual enzymatic-biosensor for simultaneous determination of glucose and cholesterol in serum and peritoneal macrophages of diabetic mice: Evaluation of the diabetes-accelerated atherosclerosis risk

    Energy Technology Data Exchange (ETDEWEB)

    Huang Qilin; An Yarui; Tang Linlin; Jiang Xiaoli; Chen Hua; Bi Wenji [Department of Chemistry, East China Normal University, Shanghai 200062 (China); Wang Zhongchuan [Department of Anorectal Surgery, Xinhua Hospital, Affiliated to School of Medicine of Shanghai Jiaotong University, Shanghai 200092 (China); Zhang Wen, E-mail: wzhang@chem.ecnu.edu.cn [Department of Chemistry, East China Normal University, Shanghai 200062 (China)

    2011-11-30

    Graphical abstract: In this paper, we reported a novel dual enzymatic-biosensor for simultaneous determination of glucose and cholesterol in serum and peritoneal macrophages (PMs) of diabetic mice to evaluate the diabetes-accelerated atherosclerosis risk. The biosensor was firstly modified with a poly-thionine (PTH) film as electron transfer mediator (ETM), then the gold nanoparticles (GNPs) were covered on the surface of PTH to act as tiny conduction centers for facilitating the electron transfer between enzymes and electrode. The schematic of the dual biosensor is shown in figure. The developed dual biosensor had good electrocatalytic activity toward the oxidations of glucose and cholesterol, exhibited a linear range from 0.008 mM to 6.0 mM for glucose with a detection limit of 2.0 {mu}M, and a linear range from 0.002 mM to 1.0 mM for cholesterol with a detection limit of 0.6 {mu}M. The results of the diabetic mice demonstrated that the cholesterol level was not changed obviously with the increase of glucose level in serum, while the cholesterol level was enhanced together with the increase of the glucose level in PMs. Previous studies have shown that the large accumulation of cholesterol in macrophage could lead to macrophage foam cell formation, the hallmark of early atherosclerosis. These findings indicated the possibility that high glucose induced by diabetes might increase the macrophage cholesterol level to further accelerate atherosclerosis development. Highlights: Black-Right-Pointing-Pointer A novel biosensor was developed to determine glucose and cholesterol simultaneously. Black-Right-Pointing-Pointer The dual enzymatic-biosensor has good selectivity and high sensitivity. Black-Right-Pointing-Pointer We determined glucose and cholesterol in the real samples of diabetic mice. Black-Right-Pointing-Pointer The results showed that high glucose might increase the macrophage cholesterol level. Black-Right-Pointing-Pointer It provided useful experimental

  8. A dual enzymatic-biosensor for simultaneous determination of glucose and cholesterol in serum and peritoneal macrophages of diabetic mice: Evaluation of the diabetes-accelerated atherosclerosis risk

    International Nuclear Information System (INIS)

    Huang Qilin; An Yarui; Tang Linlin; Jiang Xiaoli; Chen Hua; Bi Wenji; Wang Zhongchuan; Zhang Wen

    2011-01-01

    Graphical abstract: In this paper, we reported a novel dual enzymatic-biosensor for simultaneous determination of glucose and cholesterol in serum and peritoneal macrophages (PMs) of diabetic mice to evaluate the diabetes-accelerated atherosclerosis risk. The biosensor was firstly modified with a poly-thionine (PTH) film as electron transfer mediator (ETM), then the gold nanoparticles (GNPs) were covered on the surface of PTH to act as tiny conduction centers for facilitating the electron transfer between enzymes and electrode. The schematic of the dual biosensor is shown in figure. The developed dual biosensor had good electrocatalytic activity toward the oxidations of glucose and cholesterol, exhibited a linear range from 0.008 mM to 6.0 mM for glucose with a detection limit of 2.0 μM, and a linear range from 0.002 mM to 1.0 mM for cholesterol with a detection limit of 0.6 μM. The results of the diabetic mice demonstrated that the cholesterol level was not changed obviously with the increase of glucose level in serum, while the cholesterol level was enhanced together with the increase of the glucose level in PMs. Previous studies have shown that the large accumulation of cholesterol in macrophage could lead to macrophage foam cell formation, the hallmark of early atherosclerosis. These findings indicated the possibility that high glucose induced by diabetes might increase the macrophage cholesterol level to further accelerate atherosclerosis development. Highlights: ► A novel biosensor was developed to determine glucose and cholesterol simultaneously. ► The dual enzymatic-biosensor has good selectivity and high sensitivity. ► We determined glucose and cholesterol in the real samples of diabetic mice. ► The results showed that high glucose might increase the macrophage cholesterol level. ► It provided useful experimental evidences for diabetes-accelerate atherosclerosis. - Abstract: In this paper, a novel dual enzymatic-biosensor is described for

  9. Pretreating lignocellulosic biomass by the concentrated phosphoric acid plus hydrogen peroxide (PHP) for enzymatic hydrolysis: evaluating the pretreatment flexibility on feedstocks and particle sizes.

    Science.gov (United States)

    Wang, Qing; Wang, Zhanghong; Shen, Fei; Hu, Jinguang; Sun, Fubao; Lin, Lili; Yang, Gang; Zhang, Yanzong; Deng, Shihuai

    2014-08-01

    In order to seek a high-efficient pretreatment path for converting lignocellulosic feedstocks to fermentable sugars by enzymatic hydrolysis, the concentrated H₃PO₄ plus H₂O₂ (PHP) was attempted to pretreat different lignocellulosic biomass for evaluating the pretreatment flexibility on feedstocks. Meanwhile, the responses of pretreatment to particle sizes were also evaluated. When the PHP-pretreatment was employed (final H₂O₂ and H₃PO₄ concentration of 1.77% and 80.0%), 71-96% lignin and more than 95% hemicellulose in various feedstocks (agricultural residues, hardwood, softwood, bamboo, and their mixture, and garden wastes mixture) can be removed. Consequently, more than 90% glucose conversion was uniformly achieved indicating PHP greatly improved the pretreatment flexibility to different feedstocks. Moreover, when wheat straw and oak chips were PHP-pretreated with different sizes, the average glucose conversion reached 94.9% and 100% with lower coefficient of variation (7.9% and 0.0%), which implied PHP-pretreatment can significantly weaken the negative effects of feedstock sizes on subsequent conversion. Copyright © 2014 Elsevier Ltd. All rights reserved.

  10. Evaluation of the effects of a VEGFR-2 inhibitor compound on alanine aminotransferase gene expression and enzymatic activity in the rat liver.

    Science.gov (United States)

    Fuentealba, Carmen; Bera, Monali; Jessen, Bart; Sace, Fred; Stevens, Greg J; Trajkovic, Dusko; Yang, Amy H; Evering, Winston

    2011-08-17

    Traditional assessment of drug-induced hepatotoxicity includes morphological examination of the liver and evaluation of liver enzyme activity in serum. The objective of the study was to determine the origin of drug-related elevation in serum alanine aminotransferase (ALT) activity in the absence of morphologic changes in the liver by utilizing molecular and immunohistochemical techniques. Sixteen female Sprague-Dawley rats were divided into 2 groups (control and treated, n = 4 per group) and treated rats were dosed orally twice daily (400 mg/kg/day) for 7 days with a VEGFR-2 compound (AG28262), which in a previous study caused ALT elevation without morphological changes. Serum of both treated and control animals were evaluated on day 3 of treatment and at day 8. Three separate liver lobes (caudate, right medial, and left lateral) were examined for determination of ALT tissue activity, ALT gene expression and morphological changes. ALT activity was significantly (p < 0.01) elevated on day 3 and further increased on day 8. Histologic changes or increase in TUNEL and caspase3 positive cells were not observed in the liver lobes examined. ALT gene expression in the caudate lobe was significantly up-regulated by 63%. ALT expression in the left lateral lobe was not significantly affected. Statistically significant increased liver ALT enzymatic activity occurred in the caudate (96%) and right medial (41%) lobes but not in the left lateral lobe. AG28262, a VEFG-r2 inhibitor, causes an increase in serum ALT, due in part to both gene up-regulation. Differences between liver lobes may be attributable to differential distribution of blood from portal circulation. Incorporation of molecular data, such as gene and protein expression, and sampling multiple liver lobes may shed mechanistic insight to the evaluation of hepatotoxicity.

  11. Otimização da reação de interesterificação química do óleo de palma Optimization of the cemical interesterification reaction of palm oil

    Directory of Open Access Journals (Sweden)

    Renato Grimaldi

    2005-08-01

    Full Text Available The Brazilian market has been showing a growing concern with nutritional values of oil components of foods. Chemical interesterification is a promising alternative to the current processes of modifying the consistency of oils. Chemical interesterification of deodorized palm oil was studied on a laboratory scale. The best results were obtained with 0.4% MeONa and heating for 20 min at 100 °C. These conditions are based on the largest variation in triacylglycerols as compared to a control. The trisaturated values varied from 6.2 to 9.9%, showing that the consistency of the oil improved for it to be used in margarines, without the formation of trans isomers.

  12. Evaluating the Biodeterioration Enzymatic Activities of Fungal Contamination Isolated from Some Ancient Yemeni Mummies Preserved in the National Museum

    Directory of Open Access Journals (Sweden)

    Khalid Mohammed Naji

    2014-01-01

    Full Text Available Sophisticated mummification using chemical preservation was prevalent in ancient Yemeni civilization as noted in the 4th century B.C. mummies of the National Museum of Yemen, Sana’a, used in this study. Five of these mummies were used to evaluate hydrolytic enzymes produced as a result of fungal contamination. Forty-seven fungal species were isolated, thereby reflecting a high degree of contamination which may have resulted from the poor ventilation and preservation system. Aspergillus was the most common genus isolated (48.9%. Fifteen isolates exhibited ability to produce cellulase (EC; 3.2.1.4, Aspergillus candidus being the highest cellulose-producer. Pectin lyase (PL, EC; 4.2.2.2 and pectin methyl esterase (PME, EC; 3.1.1.11 were produced by Trichoderma hamatum, whereas chitinase (EC; 3.2.1.14 was produced by Aspergillus niger. Protease activity was noted by only Cladosporium herbarum. The higher activities of these fungal hydrolytic enzymes represent the major threats of biodeterioration including deteriorating linen bandages as well as the mummy bodies. Therefore, it is recommended to improve the preservation system of the mummies at the National Museum to minimize the contamination up to the lowest level and protect the mummies from biodeterioration.

  13. Studies on performance evaluation of a green plasticizer made by enzymatic esterification of furfuryl alcohol and castor oil fatty acid.

    Science.gov (United States)

    Mukherjee, Sohini; Ghosh, Mahua

    2017-02-10

    The esterification of furfuryl alcohol (FA) and castor oil fatty acid (COFA) at 3:1 molar ratio, by immobilized Candida antarctica Lipase B (NS 435 from Novozyme) in a solvent free system gave a maximum yield of 88.64% (%w/w) at 5h. Performance of the FA-COFA ester plasticized Ethyl Cellulose (EC) films were evaluated by surface morphologies, XRD analysis, mechanical properties,thermal properties, water vapor permeability and migration stability test. It was an effective plasticizer with better mechanical properties and thermal stability at the increasing concentration of FA-COFA ester (15-25%) containing EC film, than the traditional plasticizer, i.e; dibutyl phthalate (DBP) in producing good quality films. Chemical structure and the intermolecular interactions between FA-COFA ester and ethyl cellulose chains were the causative agents of these outstanding performances. Therefore, this FA-COFA ester, with significant plasticizing property, at a certain concentration, can be a substitute of DBP. Copyright © 2016 Elsevier Ltd. All rights reserved.

  14. Long-term liquid storage and reproductive evaluation of an innovative boar semen extender (Formula12®) containing a non-reducing disaccharide and an enzymatic agent.

    Science.gov (United States)

    Bresciani, Carla; Bianchera, Annalisa; Bettini, Ruggero; Buschini, Annamaria; Marchi, Laura; Cabassi, Clotilde Silvia; Sabbioni, Alberto; Righi, Federico; Mazzoni, Claudio; Parmigiani, Enrico

    2017-05-01

    There are no reports of saccharolytic enzymes being used in the preparation of formulations for animal semen extenders. In the present study, the use of an innovative semen extender (Formula12 ® ) in the long-term liquid storage of boar semen at 17°C was evaluated. The formulation included use of a disaccharide (sucrose) as the energy source precursor coupled to an enzymatic agent (invertase). The innovative extender was evaluated and compared in vitro to a commercial extender (Vitasem LD ® ) for the following variables: Total Motility (TM), Forward Progressive Motility (FPM), sperm morphology, membrane integrity, acrosome integrity, and chromatin instability. Boar sperm diluted in Formula12 ® and stored for 12 days at 17°C maintained a commercially acceptable FPM (>70%). Using the results from the in vitro study, an AI field trial was performed. A total of 170 females were inseminated (135 with Formula12 ® and 35 with Vitasem LD ® ). The pregnancy rates were 97.8% compared with 91.4%, and the farrowing rates were 96.3% compared with 88.6% when Formula12 ® and Vitasem LD ® were used, respectively. The mean number of piglets born/sow were 14.92±0.46 compared with 13.83±0.70, and the number of piglets born alive/sow were 14.07±0.46 compared with 12.12±0.70 (Pextender allowed for meeting the metabolic requirements of boar sperm during storage at 17°C. It is suggested that there was a beneficial effect on fertilizing capacity of boar sperm in the female reproductive tract with use of these technologies. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. Pollution in the urban soils of Lianyungang, China, evaluated using a pollution index, mobility of heavy metals, and enzymatic activities.

    Science.gov (United States)

    Li, Yu; Li, Hong-Guan; Liu, Fu-Cheng

    2017-01-01

    Soil samples from 16 urban sites in Lianyungang, China were collected and analyzed. A pollution index was used to assess the potential ecological risk of heavy metals and a sequential extraction procedure was used to evaluate the relative distribution of Cu, Zn, Pb, Cd, Cr, and As in exchangeable, carbonate, Fe/Mn oxide, organic/sulfide, and residual fractions. The mobility of heavy metals and urease (URE) activity, alkaline phosphatase (ALP) activity, and invertase (INV) activity of soils was determined. The results showed that the average concentrations of Cu, Zn, Pb, Cd, Cr, and As in Lianyungang soils were much higher than those in the coastal city soil background values of Jiangsu and China. Among the five studied regions (utilities, commercial, industrial, tourism, and roadside), the industrial region had the highest metal concentrations demonstrating that land use had a significant impact on the accumulation of heavy metals in Lianyungang soils. Compared to the other metals, Cd showed the highest ecological risk. According to chemical partitioning, Cu was associated with the organic/sulfides and Pb and Zn were mainly in the carbonate and the Fe/Mn oxide phase. The greatest amounts of Cd were found in exchangeable and carbonate fractions, while Cr and As were mainly in the residual fraction. Cd had the highest mobility of all metals, and the order of mobility (highest to lowest) of heavy metals in Lianyungang soils was Cd > Zn > Pb > Cu > As > Cr. Soil urease activity, alkaline phosphatase activity, and invertase activity varied considerably in different pollution degree sites. Soil enzyme activities had the lowest levels in roadside and industrial regions. Across all the soil data in the five regions, the total Cu, Zn, Pb, Cd, Cr, and As level was negatively correlated with urease activity, alkaline phosphatase activity, and invertase activity, but the relationship was not significant. In the industrial region, alkaline phosphatase activity had

  16. Photoelectrochemical enzymatic biosensors.

    Science.gov (United States)

    Zhao, Wei-Wei; Xu, Jing-Juan; Chen, Hong-Yuan

    2017-06-15

    Enzymatic biosensors have been valuable bioanalytical devices for analysis of diverse targets in disease diagnosis, biological and biomedical research, etc. Photoelectrochemical (PEC) bioanalysis is a recently emerged method that promptly becoming a subject of new research interests due to its attractive potential for future bioanalysis with high sensitivity and specificity. PEC enzymatic biosensors integrate the inherent sensitivities of PEC bioanalysis and the selectivity of enzymes and thus share their both advantages. Currently, PEC enzymatic biosensors have become a hot topic of significant research and the recent impetus has grown rapidly as demonstrated by increased research papers. Given the pace of advances in this area, this review will make a thorough discussion and survey on the fundamentals, sensing strategies, applications and the state of the art in PEC enzymatic biosensors, followed by future prospects based on our own opinions. We hope this work could provide an accessible introduction to PEC enzymatic biosensors for any scientist. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Enzymatic activity of the cellulolytic complex produced by trichoderma reesei. Enzymatic hydrolysis of cellulose

    International Nuclear Information System (INIS)

    Alfonsel Jaen, M.; Negro, M.J.; Saez, R.; Martin Moreno, C.

    1986-01-01

    The enzymatic activity characterization of the cellulolytic complex obtained from Trichoderma reese QM 9414 and the influence of the enzymatic hydrolysis conditions on the hydrolysis yield are studied. Pure cellulose and native or alkali pretreated biomass from Onopordum nervosum have been used as substrates. The values of pH, temperature, substrate concentration and enzyme-substrate ratio for the optimum activity of that complex, evaluated as glucose and reducing sugars productions, have been selected. Previous studies on enzymatic hydrolysis of O. nervosum have shown a remarkable effect of the alkaline pretreatments on the final hydrolysis yield. (author). 10 figs.; 10 refs

  18. Enzymatic activity of the cellulolytic complex produced by Trichoderma reesei. Enzymatic hydrolysis of cellulose

    International Nuclear Information System (INIS)

    Alfonsel J, M.; Negro A, M. J.; Saez A, R.; Martin M, C.

    1986-01-01

    The enzymatic activity characterization of the cellulolytic complex obtained from Trichoderma reesei QM 9414 and the influence of the enzymatic hydrolysis conditions on the hydrolysis yield are studied. Pure cellulose and native or alkali pretreated biomass Onopordum nervosum have been used as substrates. The values of pH, temperature, substrate concentration and enzyme-substrate ratio for the optimum activity of that complex, evaluated as glucose and reducing sugars production, have been selected. Previous studies on enzymatic hydrolysis of 0. nervosum have shown a remarkable effect of the alkaline pretreatments on the final hydrolysis yield. (Author) 10 refs

  19. Enzymatic synthesis of vanillin

    NARCIS (Netherlands)

    van den Heuvel, RHH; Fraaije, MW; Laane, C; van Berkel, WJH; Heuvel, Robert H.H. van den; Berkel, Willem J.H. van

    Due to increasing interest in natural vanillin, two enzymatic routes for the synthesis of vanillin were developed. The flavoprotein vanillyl alcohol oxidase (VAO) acts on a wide range of phenolic compounds and converts both creosol and vanillylamine to vanillin with high yield. The VAO-mediated

  20. Enzymatic synthesis of vanillin

    NARCIS (Netherlands)

    Heuvel, van den R.H.H.; Fraaije, M.W.; Laane, C.; Berkel, van W.J.H.

    2001-01-01

    Due to increasing interest in natural vanillin, two enzymatic routes for the synthesis of vanillin were developed. The flavoprotein vanillyl alcohol oxidase (VAO) acts on a wide range of phenolic compounds and converts both creosol and vanillylamine to vanillin with high yield. The VAO-mediated

  1. Enzymatic desulfurization of coal

    Energy Technology Data Exchange (ETDEWEB)

    Boyer, Y.N.; Crooker, S.C.; Kitchell, J.P.; Nochur, S.V.

    1991-05-16

    The overall objective of this program was to investigate the feasibility of an enzymatic desulfurization process specifically intended for organic sulfur removal from coal. Toward that end, a series of specific objectives were defined: (1) establish the feasibility of (bio)oxidative pretreatment followed by biochemical sulfate cleavage for representative sulfur-containing model compounds and coals using commercially-available enzymes; (2) investigate the potential for the isolation and selective use of enzyme preparations from coal-utilizing microbial systems for desulfurization of sulfur-containing model compounds and coals; and (3) develop a conceptual design and economic analysis of a process for enzymatic removal of organic sulfur from coal. Within the scope of this program, it was proposed to carry out a portion of each of these efforts concurrently. (VC)

  2. Enzymatic Synthesis of Psilocybin.

    Science.gov (United States)

    Fricke, Janis; Blei, Felix; Hoffmeister, Dirk

    2017-09-25

    Psilocybin is the psychotropic tryptamine-derived natural product of Psilocybe carpophores, the so-called "magic mushrooms". Although its structure has been known for 60 years, the enzymatic basis of its biosynthesis has remained obscure. We characterized four psilocybin biosynthesis enzymes, namely i) PsiD, which represents a new class of fungal l-tryptophan decarboxylases, ii) PsiK, which catalyzes the phosphotransfer step, iii) the methyltransferase PsiM, catalyzing iterative N-methyl transfer as the terminal biosynthetic step, and iv) PsiH, a monooxygenase. In a combined PsiD/PsiK/PsiM reaction, psilocybin was synthesized enzymatically in a step-economic route from 4-hydroxy-l-tryptophan. Given the renewed pharmaceutical interest in psilocybin, our results may lay the foundation for its biotechnological production. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. Ultrasonic-assisted enzymatic extraction of phenolics from broccoli (Brassica oleracea L. var. italica) inflorescences and evaluation of antioxidant activity in vitro.

    Science.gov (United States)

    Wu, Hao; Zhu, Junxiang; Yang, Long; Wang, Ran; Wang, Chengrong

    2015-06-01

    An efficient ultrasonic-assisted enzymatic extraction technique was applied to extracting phenolics from broccoli inflorescences without organic solvents. The synergistic model of enzymolysis and ultrasonication simultaneously was selected, and the enzyme combination was optimized by orthogonal test: cellulase 7.5 mg/g FW (fresh weight), pectinase 10 mg/g FW, and papain 1.0 mg/g FW. The operating parameters in ultrasonic-assisted enzymatic extraction were optimized with response surface methodology using Box-Behnken design. The optimal extraction conditions were as follows: ultrasonic power, 440 W; liquid to material ratio, 7.0:1 mL/g; pH value of 6.0 at 54.5 ℃ for 10 min. Under these conditions, the extraction yield of phenolics achieved 1.816 ± 0.0187 mg gallic acid equivalents/gram FW. The free radical scavenging activity of ultrasonic-assisted enzymatic extraction extracts was determined by 1,1-diphenyl-2-picrylhydrazyl·assay with EC50 values of 0.25, and total antioxidant activity was determined by ferric reducing antioxidant power assay with ferric reducing antioxidant power value of 0.998 mmol FeSO4/g compared with the referential ascorbic acid of 1.184 mmol FeSO4/g. © The Author(s) 2014 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.

  4. Antioxidant Potential and Modulatory Effects of Restructured Lipids from the Amazonian Palms on Liver Cells

    Directory of Open Access Journals (Sweden)

    Andrea de Oliveira Falcão

    2017-01-01

    Full Text Available Enzymatic interesterification is used to manipulate oil and fat in order to obtain improved restructured lipids with desired technological properties. However, with raw materials containing significant amounts of bioactive compounds, the influence of this enzymatic process on the bioactivity of the final product is still not clear. Thus, the aim of this study is to evaluate the antioxidant potential and modulatory effects of two raw materials from the Amazonian area, buriti oil and murumuru fat, before and after lipase interesterification, on human hepatoma cells (HepG2. The results indicate that minor bioactive compounds naturally found in the raw materials and their antioxidant capacity are preserved after enzymatic interesterification, and that the restructured lipids modulate HepG2 endogenous antioxidant enzyme.

  5. Evaluation of Biological and Enzymatic Activity of Soil in a Tropical Dry Forest: Desierto de la Tatacoa (Colombia) with Potential in Mars Terraforming and Other Similar Planets

    Science.gov (United States)

    Moreno Moreno, A. N.

    2009-12-01

    Desierto de la Tatacoa has been determined to be a tropical dry forest bioma, which is located at 3° 13" N 75° 13" W. It has a hot thermal floor with 440 msnm of altitude; it has a daily average of 28° C, and a maximum of 40° C, Its annual rainfall total can be upwards of 1250 mm. Its solar sheen has a daily average of 5.8 hours and its relative humidity is between 60% and 65%. Therefore, the life forms presents are very scant, and in certain places, almost void. It was realized a completely random sampling of soil from its surface down to 6 inches deep, of zones without vegetation and with soils highly loaded by oxides of iron in order to determine the number of microorganisms per gram and its subsequent identification. It was measured the soil basal respiration. Besides, it was determined enzymatic activity (catalase, dehydrogenase, phosphatase and urease). Starting with the obtained results, it is developes an alternative towards the study of soil genesis in Mars in particular, and recommendations for same process in other planets. Although the information found in the experiments already realized in Martian soil they demonstrate that doesnt exist any enzymatic activity, the knowledge of the same topic in the soil is proposed as an alternative to problems like carbonic fixing of the dense Martian atmosphere of CO2, the degradation of inorganic compounds amongst other in order to prepare the substratum for later colonization by some life form.

  6. Interesterification of engkabang (Shorea macrophylla) fat--canola oil blend with lipase from Candida antarctica to simulate the properties of lard.

    Science.gov (United States)

    Illiyin, Mohamed Roslan Nur; Marikkar, Jalaldeen Mohamed Nazrim; Loke, Mei Key; Shuhaimi, Musthafa; Mahiran, Basri; Miskandar, Mat Saari

    2014-01-01

    A study was carried out to compare the composition and thermal properties of lard (LD) and engkabang fat (EF) - canola oil (CaO) blend interesterified with Candida antartica lipase (C. antartica). A fat blend EF-4 (40% EF in CaO) was prepared and interesterified using C. antartica lipase at 60°C for different time intervals (6 h, 12 h and 24 h) with 200 rpm agitation. The fat blends before and after interesterification were compared to LD with respect to their slip melting points (SMP), fatty acid and triacyglycerol (TAG) compositions, melting, solidification and polymorphic properties. Result showed that the slip melting point (SMP) of the fat blend interesterified for 6 h was the closest to that of LD. The solid fat content (SFC) values of fat blends interesterified for 12 and 24 h were found to become equal to those of LD within the temperature range of 0 to 20°C. In addition, all three interesterified blends had SFC values similar to those of LD within the temperature range of 30-40°C. According to thermal analysis, the transition of the fat blend interesterified for 24 h appearing at -2.39°C was similar to the low melting thermal transition of LD and the transition of the fat blend interesterified for 12 h appearing at 26.25°C was similar to the high melting thermal transition of LD. However, there is no compatibility between LD and all three interesterified blends with regard to polymorphic behaviour.

  7. Enzymatic synthesis of {sup 125/131}I labeled 8-hydroxyquinoline glucuronide and in vitro/in vivo evaluation of biological influence

    Energy Technology Data Exchange (ETDEWEB)

    Yesilagac, Reyhan [Ege University, Institute of Nuclear Science, 35100 Bornova, Izmir (Turkey); Unak, Perihan, E-mail: perihan.unak@ege.edu.t [Ege University, Institute of Nuclear Science, 35100 Bornova, Izmir (Turkey); Medine, E. Ilker; Ichedef, Cigdem A. [Ege University, Institute of Nuclear Science, 35100 Bornova, Izmir (Turkey); Ertay, Turkan [Dokuz Eyluel University, Medical School, Department of Nuclear Medicine, Inciralti, Izmir (Turkey); Mueftueler, F.Z. Biber [Ege University, Institute of Nuclear Science, 35100 Bornova, Izmir (Turkey)

    2011-02-15

    8-Hydroxyquinoline (8-OHQ) is a long-known molecule which due to its metal-complexation ability is frequently used for analysis. It is also called oxine. Oxine and derivatives have been investigated to process antitumor and antimicrobial activities. 8-Hydroxyquinolyl-glucuronide (8-OHQ-Glu) was enzymatically synthesized using microsome preparates separated from Hutu-80 cells, labeled with {sup 125}I to perform a radionuclide labeled prodrug and investigated of its biological affinities on Hutu-80 (human duodenum intestinal adenocarcinoma), Caco-2 (human colorectal adenocarcinoma), Detroit 562 (human pharynx adenocarcinoma) cells and ACBRI 519 (primary human small intestine epithelial cells) in this work. UDP-glucuronyl transferase (UDPGT) rich microsome preparates, which are used for glucuronidation in enzymatic synthesis, were extracted from Hutu-80 cells. 8-OHQ-Glu components were labeled using iodogen method with {sup 125}I and {sup 131}I. Structural analyses were performed with LC/MS/MS, {sup 1}H NMR and {sup 13}C-MMR for identify and measure chemical constituents. Results confirmed expected molecular structure. 8-OHQ-Glu could successfully radioiodinated with {sup 125/131}I according to iodogen method. {sup 125}I-8-OHQ-glucuronide incorporated with human gastrointestinal cancer cells such as Detroit-562 (human pharynx adenocarcinoma) (12.6%), Caco-2 (human colorectal adenocarcinoma) (7.8%), Hutu- 80 (human duodenum intestinal adenocarcinoma) (9.5%) and ACBRI 519 (primary human small intestine epithelial cells) (6.40%). {sup 131}I-8-OHQ-Glu was tested in mice bearing subcutaneously implanted Caco-2 colorectal adenocarcinoma cells. The results demonstrated that radioiodinated 8-OHQ-Glu may be promising anticancer prodrug.

  8. Application of cavitation system to accelerate aqueous enzymatic extraction of seed oil from Cucurbita pepo L. and evaluation of hypoglycemic effect.

    Science.gov (United States)

    Li, Xiao-Juan; Li, Zhu-Gang; Wang, Xun; Han, Jun-Yan; Zhang, Bo; Fu, Yu-Jie; Zhao, Chun-Jian

    2016-12-01

    Cavitation-accelerated aqueous enzymatic extraction (CAEE) of seed oil from Cucurbita pepo was performed. An enzyme cocktail comprised of cellulose, pectinase and proteinase can work synergistically in releasing the oil. The CAEE extraction conditions were optimized by a Plackett-Burman design followed by a central composite methodology. A maximal extraction yield of 58.06% was achieved under optimal conditions of vacuum degree -0.07, enzyme amount 1.05% and extraction time 69min. As compared to soxhlet extraction (SE)-derived oil, CAEE-derived oil exhibited similar physical properties and better oxidation stability. In addition, chemical composition analyzing showed that the content of linoleic acid obtained by CAEE (47.67%) was higher than that of SE (44.51%). Moreover, the IC50 of oil obtained by CAEE and SE, as measured by α-amylase inhibition assay, were 40.68μg/mL and 45.46μg/mL. All results suggest that CAEE represents an excellent alternative protocol for production of oil from oil-bearing materials. Copyright © 2016 Elsevier Ltd. All rights reserved.

  9. Microwave-assisted aqueous enzymatic extraction of oil from pumpkin seeds and evaluation of its physicochemical properties, fatty acid compositions and antioxidant activities.

    Science.gov (United States)

    Jiao, Jiao; Li, Zhu-Gang; Gai, Qing-Yan; Li, Xiao-Juan; Wei, Fu-Yao; Fu, Yu-Jie; Ma, Wei

    2014-03-15

    Microwave-assisted aqueous enzymatic extraction (MAAEE) of pumpkin seed oil was performed in this study. An enzyme cocktail comprised of cellulase, pectinase and proteinase (w/w/w) was found to be the most effective in releasing oils. The highest oil recovery of 64.17% was achieved under optimal conditions of enzyme concentration (1.4%, w/w), temperature (44°C), time (66 min) and irradiation power (419W). Moreover, there were no significant variations in physicochemical properties of MAAEE-extracted oil (MAAEEO) and Soxhlet-extracted oil (SEO), but MAAEEO exhibited better oxidation stability. Additionally, MAAEEO had a higher content of linoleic acid (57.33%) than SEO (53.72%), and it showed stronger antioxidant activities with the IC50 values 123.93 and 152.84, mg/mL, according to DPPH radical scavenging assay and β-carotene/linoleic acid bleaching test. SEM results illustrated the destruction of cell walls and membranes by MAAEE. MAAEE is, therefore, a promising and environmental-friendly technique for oil extraction in the food industry. Copyright © 2013 Elsevier Ltd. All rights reserved.

  10. Microplate-Based Evaluation of the Sugar Yield from Giant Reed, Giant Miscanthus and Switchgrass after Mild Chemical Pre-Treatments and Hydrolysis with Tailored Trichoderma Enzymatic Blends.

    Science.gov (United States)

    Cianchetta, Stefano; Bregoli, Luca; Galletti, Stefania

    2017-11-01

    Giant reed, miscanthus, and switchgrass are considered prominent lignocellulosic feedstocks to obtain fermentable sugars for biofuel production. The bioconversion into sugars requires a delignifying pre-treatment step followed by hydrolysis with cellulase and other accessory enzymes like xylanase, especially in the case of alkali pre-treatments, which retain the hemicellulose fraction. Blends richer in accessory enzymes than commercial mix can be obtained growing fungi on feedstock-based substrates, thus ten selected Trichoderma isolates, including the hypercellulolytic strain Trichoderma reesei Rut-C30, were grown on giant reed, miscanthus, or switchgrass-based substrates. The produced enzymes were used to saccharify the corresponding feedstocks, compared to a commercial enzymatic mix (6 FPU/g). Feedstocks were acid (H 2 SO 4 0.2-2%, w/v) or alkali (NaOH 0.02-0.2%, w/v) pre-treated. A microplate-based approach was chosen for most of the experimental steps due to the large number of samples. The highest bioconversion was generally obtained with Trichoderma harzianum Or4/99 enzymes (78, 89, and 94% final sugar yields at 48 h for giant reed, miscanthus, and switchgrass, respectively), with significant increases compared to the commercial mix, especially with alkaline pre-treatments. The differences in bioconversion yields were only partially caused by xylanases (maximum R 2  = 0.5), indicating a role for other accessory enzymes.

  11. Development and evaluation of a self-cleaning custom-built auto sampler controlled by a low-cost RaspberryPi microcomputer for online enzymatic activity measurements.

    Science.gov (United States)

    Stadler, Philipp; Farnleitner, Andreas H; Zessner, Matthias

    2017-01-01

    A fully automated on-site device (SAMP-FIL) that enables water sampling with simultaneous filtration and effective cleaning procedures of the device's components was developed and field-tested. The SAMP-FIL was custom-built using commercially available components and was controlled by a RaspberryPi single-board computer operating open-source software. SAMP-FIL was designed for sample pre-treatment with minimal sample alteration to meet the requirements of on-site measurement devices that cannot handle coarse suspended solids within the measurement procedure or cycle. A highly effective cleaning procedure provides a fresh and minimally altered sample for the connected measurement device. The construction and programmed software facilitates the use of SAMP-FIL for different connected measurement devices. The SAMP-FIL sample pretreatment was tested for over one year for rapid and on-site enzymatic activity (beta-d-glucuronidase, GLUC) determination (BACTcontrol) in sediment-laden stream water. The formerly used proprietary sampling set-up was assumed to lead to significant damping of the measurement signal due to its susceptibility to clogging, debris accumulation and bio-film accumulation. The implementation of SAMP-FIL considerably increased the error-free running time and measurement accuracy of BACTcontrol devices. This paper describes how low-cost microcomputers, such as the RaspberryPi, can be used by operators to substantially improve established measuring systems via effective sampling devices. Furthermore, the results of this study highlight the importance of adequate sample pretreatment for the quality of on-site measurements. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

  12. Enzymatic and viability RT-qPCR assays for evaluation of enterovirus, hepatitis A virus and norovirus inactivation: Implications for public health risk assessment.

    Science.gov (United States)

    Monteiro, S; Santos, R

    2018-04-01

    To assess the potential of a viability dye and an enzymatic reverse transcription quantitative PCR (RT-qPCR) pretreatment to discriminate between infectious and noninfectious enteric viruses. Enterovirus (EntV), norovirus (NoV) GII.4 and hepatitis A virus (HAV) were inactivated at 95°C for 10 min, and four methods were used to compare the efficiency of inactivation: (i) cell culture plaque assay for HAV and EntV, (ii) RT-qPCR alone, (iii) RT-qPCR assay preceded by RNase treatment, and (iv) pretreatment with a viability dye (reagent D (RD)) followed by RT-qPCR. In addition, heat-inactivated NoV was treated with RD coupled with surfactants to increase the efficiency of the viability dye. No treatment was able to completely discriminate infectious from noninfectious viruses. RD-RT-qPCR reduced more efficiently the detection of noninfectious viruses with little to no removal observed with RNase. RD-RT-qPCR method was the closest to cell culture assay. The combination of surfactants and RD did not show relevant improvements on the removal of inactivated viruses signal compared with viability RT-qPCR, with the exception of Triton X-100. The use of surfactant/RD-RT-qPCR, although not being able to completely remove the signal from noninfectious viral particles, yielded a better estimation of viral infectivity. Surfactant/RD-RT-qPCR may be an advantageous tool for a better detection of infectious viruses with potential significant impact in the risk assessment of the presence of enteric viruses. © 2017 The Society for Applied Microbiology.

  13. Process technology for multi-enzymatic reaction systems

    DEFF Research Database (Denmark)

    Xue, Rui; Woodley, John M.

    2012-01-01

    In recent years, biocatalysis has started to provide an important green tool in synthetic organic chemistry. Currently, the idea of using multi-enzymatic systems for industrial production of chemical compounds becomes increasingly attractive. Recent examples demonstrate the potential of enzymatic...... synthesis and fermentation as an alternative to chemical-catalysis for the production of pharmaceuticals and fine chemicals. In particular, the use of multiple enzymes is of special interest. However, many challenges remain in the scale-up of a multi-enzymatic system. This review summarizes and discusses...... the technology options and strategies that are available for the development of multi-enzymatic processes. Some engineering tools, including kinetic models and operating windows, for developing and evaluating such processes are also introduced....

  14. Biosensing strategies based on enzymatic reactions and nanoparticles.

    Science.gov (United States)

    Díez-Buitrago, Beatriz; Briz, Nerea; Liz-Marzán, Luis M; Pavlov, Valeri

    2018-04-16

    Enzymes are pivotal elements in bioanalysis due to their specificity and extremely high catalytic activity. The sensitivity of bioanalytical assays depends mainly on the capacity of an observer to detect the product(s) of a biocatalytic reaction. Both natural and artificial compounds have been traditionally used to evaluate enzymatic activities. The drawbacks of chromogenic and fluorogenic organic enzymatic substrates are their high cost and low stability, resulting in high background signals. We review here state of the art assays in the detection of enzymatic activities using recent advances in nanoscience. Novel methods based on the use of nanoparticles lead to increased sensitivity and decreased costs for bioanalysis based on enzymes as recognition elements and signal amplifiers in Enzyme-Linked Immunosorbent Assays (ELISA). Novel approaches toward the detection of enzymatic activities are based on biocatalytic synthesis, modulation, etching, and aggregation of nanoparticles under physiological conditions.

  15. Poly(ethylene oxide monomethyl ether)- block-poly(propylene succinate) Nanoparticles: Synthesis and Characterization, Enzymatic and Cellular Degradation, Micellar Solubilization of Paclitaxel, and in Vitro and in Vivo Evaluation.

    Science.gov (United States)

    Jäger, Alessandro; Jäger, Eliézer; Syrová, Zdeňka; Mazel, Tomas; Kováčik, Lubomír; Raška, Ivan; Höcherl, Anita; Kučka, Jan; Konefal, Rafal; Humajova, Jana; Poučková, Pavla; Štěpánek, Petr; Hrubý, Martin

    2018-04-11

    Polyester-based nanostructures are widely studied as drug-delivery systems due to their biocompatibility and biodegradability. They are already used in the clinic. In this work, we describe a new and simple biodegradable and biocompatible system as the Food and Drug Administration approved polyesters (poly-ε-caprolactone, polylactic acid, and poly(lactic- co-glycolic acid)) for the delivery of the anticancer drug paclitaxel (PTX) as a model drug. A hydrophobic polyester, poly(propylene succinate) (PPS), was prepared from a nontoxic alcohol (propylene glycol) and monomer from the Krebs's cycle (succinic acid) in two steps via esterification and melt polycondensation. Furthermore, their amphiphilic block copolyester, poly(ethylene oxide monomethyl ether)- block-poly(propylene succinate) (mPEO- b-PPS), was prepared by three steps via esterification followed by melt polycondensation and the addition of mPEO to the PPS macromolecules. Analysis of the in vitro cellular behavior of the prepared nanoparticle carriers (NPs) (enzymatic degradation, uptake, localization, and fluorescence resonance energy-transfer pair degradation studies) was performed by fluorescence studies. PTX was loaded to the NPs of variable sizes (30, 70, and 150 nm), and their in vitro release was evaluated in different cell models and compared with commercial PTX formulations. The mPEO- b-PPS copolymer analysis displays glass transition temperature hydrolysis during transport in bloodstream, and simultaneous enzymatic degradability after uptake into the cells. The detailed cytotoxicity in vitro and in vivo tumor efficacy studies have shown the superior efficacy of the NPs compared with PTX and PTX commercial formulations.

  16. Determination of SFC, FFA, and equivalent reaction time for enzymatically interestified oils using NIRS

    DEFF Research Database (Denmark)

    Houmøller, Lars P.; Kristensen, Dorthe; Rosager, Helle

    2007-01-01

    The use of near infrared spectroscopy (NIRS) for rapid determination of the degree of interesterification of blends of palm stearin, coconut oil, and rapeseed oil obtained using an immobilized Thermomyces lanuginosa lipase at 70 ◦C was investigated. Interesterification was carried out by applying...... that NIRS could be used to replace the traditional methods for determining FFA and SFC in vegetable oils.It was possible to monitor the activity of the immobilized enzyme for interesterification of margarine oils by predicting the equivalent reaction time in a batch reactor from NIR spectra. Root mean...... square errors of prediction for two different oil blends interesterified for 300 and 170 min were 21 and 12 min, respectively....

  17. Physico-chemical properties of Moringa oleifera seed oil enzymatically interesterified with palm stearin and palm kernel oil and its potential application in food.

    Science.gov (United States)

    Dollah, Sarafhana; Abdulkarim, Sabo Mohammed; Ahmad, Siti Hajar; Khoramnia, Anahita; Mohd Ghazali, Hasanah

    2016-08-01

    High oleic acid Moringa oleifera seed oil (MoO) has been rarely applied in food products due to the low melting point and lack of plasticity. Enzymatic interesterification (EIE) of MoO with palm stearin (PS) and palm kernel oil (PKO) could yield harder fat stocks that may impart desirable nutritional and physical properties. Blends of MoO and PS or PKO were examined for triacylglycerol (TAG) composition, thermal properties and solid fat content (SFC). EIE caused rearrangement of TAGs, reduction of U3 and increase of U2 S in MoO/PS blends while reduction of U3 and S3 following increase of S2 U and U2 S in MoO/PKO blends (U, unsaturated and S, saturated fatty acids). SFC measurements revealed a wide range of plasticity, enhancements of spreadability, mouthfeel and cooling effect for interesterified MoO/PS, indicating the possible application of these blends in margarines. However, interesterified MoO/PKO was not suitable in margarine application, while ice-cream may be formulated from these blends. A soft margarine formulated from MoO/PS 70:30 revealed high oxidative stability during 8 weeks storage with no significant changes in peroxide and p-anisidine values. EIE of fats with MoO allowed nutritional and oxidative stable plastic fats to be obtained, suitable for possible use in industrial food applications. © 2015 Society of Chemical Industry. © 2015 Society of Chemical Industry.

  18. Enzymatic Browning: a practical class

    Directory of Open Access Journals (Sweden)

    Maria Teresa Pedrosa Silva Clerici

    2014-10-01

    Full Text Available This paper presents a practical class about the enzymes polyphenol oxidases, which have been shown to be responsible for the enzymatic browning of fruits and vegetables. Vegetables samples were submitted to enzymatic inactivation process with chemical reagents, as well as by bleaching methods of applying heat by conventional oven and microwave oven. Process efficiency was assessed qualitatively by both observing the guaiacol peroxidase activity and after the storage period under refrigeration or freezing. The practical results obtained in this class allow exploring multidisciplinary knowledge in food science, with practical applications in everyday life.

  19. Kinetic modelling of enzymatic starch hydrolysis

    NARCIS (Netherlands)

    Bednarska, K.A.

    2015-01-01

    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. Enzymatic hydrolsis of pretreated rice straw

    Energy Technology Data Exchange (ETDEWEB)

    Vlasenko, E.Y.; Shoemaker, S.P. [California Inst. of Food and Agricultural Research, Davis, CA (United States); Ding, H. [California Univ., Davis (Canada). Dept. of Food Science and Technology; Labavitch, J.M. [California Univ., Davis, CA (United States). Dept. of Pomology

    1997-02-01

    California rice straw is being evaluated as a feedstock for production of power and fuel. This paper examines the initial steps in the process: pretreatment of rice straw and enzymatic hydrolysis of the polysaccharides in the pretreated material to soluble sugars. Rice straw was subjected to three distinct pretreatment procedures: acid-catalyzed steam explosion (Swan Biomass Company), acid hydrolysis (U.S. DOE National Renewable Energy Laboratory), and ammonia fiber explosion or AFEX (Texas A and M University). Standard conditions for each pretreatment were used, but none was optimized for rice straw specifically. Six commercial cellulases, products of Genencor International (USA), Novo (Denmark), Iogen (Canada) and Fermtech (Russia) were used for hydrolysis. The Swan- and the acid-pretreatments effectively removed hemicellulose from rice straw, providing high yields of fermentable sugars. The AFEX-pretreatment was distinctly different from other pretreatments in that it did not significantly solubilize hemicellulose. All three pretreatment procedures substantially increased enzymatic digestibility of rice straw. Three commercial Trichoderma-reesei-derived enzyme preparations: Cellulase 100L (Iogen), Spezyme CP (Genencor), and Al (Fermtech), were more active on pretreated rice straw compared than others tested. Conditions for hydrolysis of rice straw using Cellulase 100L were evaluated. The supplementation of this enzyme preparation with cellobiase (Novozyme 188) significantly improved the parameters of hydrolysis for the Swan- and the acid-pretreated materials, but did not affect the hydrolysis of the AFEX-pretreated rice straw. (Author)

  1. The enzymatic determination of starch in food, feed and raw materials of the starch industry

    NARCIS (Netherlands)

    Brunt, K.; Sanders, P.; Rozema, T.

    1998-01-01

    An enzymatic starch determination which can be used for the analysis of starch in a very broad range of different samples is evaluated, ranging from starch in plants, feed and food to industrial applications as starch in starch. The method is based on a complete enzymatic conversion of the starch

  2. Antipathy of Trichoderma against Sclerotium rolfsii Sacc.: Evaluation of Cell Wall-Degrading Enzymatic Activities and Molecular Diversity Analysis of Antagonists.

    Science.gov (United States)

    Hirpara, Darshna G; Gajera, Harsukh P; Hirpara, Hitesh Z; Golakiya, Balubhai A

    2017-01-01

    The fungus Trichoderma is a teleomorph of the Hypocrea genus and associated with biological control of plant diseases. The microscopic, biochemical, and molecular characterization of Trichoderma was carried out and evaluated for in vitro antagonistic activity against the fungal pathogen Sclerotium rolfsii causing stem rot disease in groundnut. In total, 11 isolates of Trichoderma were examined for antagonism at 6 and 12 days after inoculation (DAI). Out of 11, T. virens NBAII Tvs12 evidenced the highest (87.91%) growth inhibition of the test pathogen followed by T. koningii MTCC 796 (67.03%), T. viride NBAII Tv23 (63.74%), and T. harzianum NBAII Th1 (60.44%). Strong mycoparasitism was observed in the best antagonist Tvs12 strain during 6-12 DAI. The specific activity of cell wall-degrading enzymes - chitinase and β-1,3-glucanase - was positively correlated with growth inhibition of the test pathogen. In total, 18 simple sequence repeat (SSR) polymorphisms were reported to amplify 202 alleles across 11 Trichoderma isolates. The average polymorphism information content for SSR markers was found to be 0.80. The best antagonist Tvs 12 was identified with 7 unique SSR alleles amplified by 5 SSR markers. Clustering patterns of 11 Trichoderma strains showed the best antagonist T. virens NBAII Tvs 12 outgrouped with a minimum 3% similarity from the rest of Trichoderma. © 2017 S. Karger AG, Basel.

  3. Therapeutic effectiveness of a new enzymatic bleaching dentifrice.

    Science.gov (United States)

    Forner, Leopaldo; Amengual, José; Liena, Carmen; Riutord, Pere

    2012-01-01

    Research into bleaching focuses on new products in order to minimize undesirable effects. This study evaluated the bleaching effectiveness of a new enzymatic-activated dentifrice. A total of 20 volunteers were bleached with a dentifrice containing 5% lactoperoxidase and 3% carbamide peroxide applied three times a day for two minutes over 21 days. Color was recorded before and after the treatment using a spectrophotometer. CIELAB differences were calculated before and after treatment using the paired t test (P whitening teeth. Enzymatic dental bleaching is able to increase the efficiency of low concentration peroxides, reducing the potential risk of peroxides on oral tissues.

  4. Moving towards a Competitive Fully Enzymatic Biodiesel Process

    Directory of Open Access Journals (Sweden)

    Silvia Cesarini

    2015-06-01

    Full Text Available Enzymatic biodiesel synthesis can solve several problems posed by the alkaline-catalyzed transesterification but it has the drawback of being too expensive to be considered competitive. Costs can be reduced by lipase improvement, use of unrefined oils, evaluation of soluble/immobilized lipase preparations, and by combination of phospholipases with a soluble lipase for biodiesel production in a single step. As shown here, convenient natural tools have been developed that allow synthesis of high quality FAMEs (EN14214 from unrefined oils in a completely enzymatic single-step process, making it fully competitive.

  5. Enzymatic Processes in Marine Biotechnology.

    Science.gov (United States)

    Trincone, Antonio

    2017-03-25

    In previous review articles the attention of the biocatalytically oriented scientific community towards the marine environment as a source of biocatalysts focused on the habitat-related properties of marine enzymes. Updates have already appeared in the literature, including marine examples of oxidoreductases, hydrolases, transferases, isomerases, ligases, and lyases ready for food and pharmaceutical applications. Here a new approach for searching the literature and presenting a more refined analysis is adopted with respect to previous surveys, centering the attention on the enzymatic process rather than on a single novel activity. Fields of applications are easily individuated: (i) the biorefinery value-chain, where the provision of biomass is one of the most important aspects, with aquaculture as the prominent sector; (ii) the food industry, where the interest in the marine domain is similarly developed to deal with the enzymatic procedures adopted in food manipulation; (iii) the selective and easy extraction/modification of structurally complex marine molecules, where enzymatic treatments are a recognized tool to improve efficiency and selectivity; and (iv) marine biomarkers and derived applications (bioremediation) in pollution monitoring are also included in that these studies could be of high significance for the appreciation of marine bioprocesses.

  6. Malondialdehyde level and some enzymatic activities in subclinical ...

    African Journals Online (AJOL)

    The purpose of this study was to evaluate the changes occurring in milk malondialdehyde (MDA) level and some enzymatic activities as a result of subclinical mastitis (SCM) in dairy cows. A total of 124 milk samples were collected from 124 lactating cows from the same herd in the period between the 2nd week after calving ...

  7. Enzymatic transesterification of used frying oils

    Energy Technology Data Exchange (ETDEWEB)

    Kovacs, S.; Hancsok, J. (Univ. of Pannonia, Veszprem (HU)), Email: hancsokj@almos.uni-pannon.hu

    2009-07-01

    The research of converting used frying oils to less harmful products with much higher value was forced by environmental, human biological and economical reasons. One possible pathway of the transformation is the enzymatic transesterification. Through the research work used frying oils (UFO) and sunflower oils (SO) from different origins were first properly pre-treated. Then the previously mentioned feeds and different mixtures of them were transesterified in the presence of Novozym 435 enzyme catalyst under different process conditions. Characteristics of the produced methyl esters were evaluated according to the requirements of EN 14214:2009 standard. We determined that the transesterification of used frying oils is not expediential in the presence of enzyme catalyst because the significant decreasing of catalyst activity. We have found proper UFO and SO mixtures and combination of process conditions (pressure: atmospheric, temperature: 54 +-1 deg C; methanol to triglyceride molar ratio: 4:1; reaction time: 16 hours) resulting in high (>90 %) yield of monoesters. We clearly established that the best results through the enzymatic transesterification were obtained with the improved sunflower oils containing the highest amount (>88 %) of oleic acid and the used frying oils originated from this source. (orig.)

  8. Avaliação do tempo de secagem e da atividade de óxido-redutases de yacon (Smallanthus sonchifolius sob tratamento químico Drying evaluation time and yacon (Smallanthus sonchifolius enzymatic activity inhibition under chemical treatment

    Directory of Open Access Journals (Sweden)

    Vivianne Montarroyos Padilha

    2009-10-01

    project aimed to evaluate the use of chemical agents in yacon processing to obtain flour in a way that inhibits enzymatic darkening of the product besides determining the enzymatic activity in these treatments. Samples of yacon without chemical inhibitions, yacon treated with 1.0g 100g-1 calcium chloride for 30 seconds and yacon treated with 0.5g 100g-1 potassium metabisulfite for 5 minutes were dried at 55oC in a ventilated greenhouse and the proportions of humidity and drying curves were determined. The peroxidase activities and polyphenol oxidase enzymes were checked before and after being dried with an enzymatic darkening possible biochemist marker of this tubercle. Regarding humidity Parameter all the three treatment were equivalent, but treatment 2 (calcium chloride reduced the humidity in lower time. Before and after the thermal treatment the enzymatic activity was higher in treatment 3 (potassium metabisulfite. The thermal action did not inhibit completely polyphenol oxidase and peroxidase. The treatment with calcium chloride at 1.0g 100g-1 for 30 minutes to obtain yacon flour was the one with better result despite the fact that it did not inhibit completely peroxidase and polyphenol oxidase enzymes activity, offering a better drying time better material of row firmness , facilitating the process for obtaining the meal.

  9. Enzymatic activity of the cellulolytic complex produced by Trichoderma reesei. Enzymatic hydrolysis of cellulose; Actividad enzimatica del complejo celulolitico producido por Trichoderma reesei. Hidrolisis enzimatica de la celulosa

    Energy Technology Data Exchange (ETDEWEB)

    Alfonsel, M; Negro, M J; Saez, R; Martin, C

    1986-07-01

    The enzymatic activity characterization of the cellulolytic complex obtained from Trichoderma reesei QM 9414 and the influence of the enzymatic hydrolysis conditions on the hydrolysis yield are studied. Pure cellulose and native or alkali pretreated biomass Onopordum nervosum have been used as substrates. The values of pH, temperature, substrate concentration and enzyme-substrate ratio for the optimum activity of that complex, evaluated as glucose and reducing sugars production, have been selected. Previous studies on enzymatic hydrolysis of 0. nervosum have shown a remarkable effect of the alkaline pretreatments on the final hydrolysis yield. (Author) 10 refs.

  10. Enzymatic production of ceramide from sphingomyelin

    DEFF Research Database (Denmark)

    Zhang, Long; Hellgren, Lars; Xu, Xuebing

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

  11. Pretreatment and enzymatic hydrolysis of lignocellulosic biomass

    Science.gov (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

  12. Evaluation of the Protective Role of Vitamin C on the Metabolic and Enzymatic Activities of the Liver in the Male Rats After Exposure to 2.45 GHz Of Wi-Fi Routers

    Directory of Open Access Journals (Sweden)

    Shekoohi-Shooli F.

    2016-09-01

    Full Text Available Background: The use of devices emitted microwave radiation such as mobile phones, wireless fidelity (Wi-Fi routers, etc. is increased rapidly. It has caused a great concern; the researchers should identify its effects on people’s health. We evaluated the protective role of Vitamin C on the metabolic and enzymatic activities of the liver after exposure to Wi-Fi routers. Material and Methods: 70 male Wistar rats weighing 200-250 g were randomly divided into 7 groups (10 rats in each group.The first stage one –day test: Group A (received vitamin C 250 mg/kg/day orally together with 8- hour/day Wi-Fi exposure. Group B (exposed to Wi-Fi radiation. Group C (received vitamin C. Group D or Control (was neither exposed to radiation of Wi-Fi modem nor did receive vitamin C. The second phase of experiment had done for five consecutive days. It involved Group E (received vitamin C, Group F (exposed to Wi-Fi radiation, Group G (received vitamin C together with Wi-Fi radiation. The distance between animals’ restrainers was 20 cm away from the router antenna. Finally, blood samples were collected and assayed the level of hepatic enzymes including alkaline phosphatase(ALP, alanine amino transferase(ALT aspartate amino transferase (ASL, gamma glutamyl transferase (GGT and the concentration of Blood Glucose, Cholesterol , Triglyceride(TG,High density lipoprotein (HDLand low density lipoprotein (LDL. Results: Data obtained from the One day test showed an increase in concentration of blood glucose, decrease in Triglyceride level and GGT factor (P<0.05, however no observed significant difference on the Cholesterol , HDL , LDL level and hepatic enzymes activities in compare to control group. Groups of the five-day test showed reduction in the amount of blood glucose, elevation of cholesterol level and LDL relative to control group(P<0.05. Conclusion: WiFi exposure may exert alternations on the metabolic parameters and hepatic enzymes activities through stress

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

    DEFF Research Database (Denmark)

    Prunescu, Remus Mihail; Sin, Gürkan

    2013-01-01

    The enzymatic hydrolysis process is one of the key steps in second generation biofuel production. After being thermally pretreated, the lignocellulosic material is liquefied by enzymes prior to fermentation. The scope of this paper is to evaluate a dynamic model of the hydrolysis process...... on a demonstration scale reactor. The following novel features are included: the application of the Convection–Diffusion–Reaction equation to a hydrolysis reactor to assess transport and mixing effects; the extension of a competitive kinetic model with enzymatic pH dependency and hemicellulose hydrolysis......; a comprehensive pH model; and viscosity estimations during the course of reaction. The model is evaluated against real data extracted from a demonstration scale biorefinery throughout several days of operation. All measurements are within predictions uncertainty and, therefore, the model constitutes a valuable...

  14. Physiological and enzymatic analyses of pineapple subjected to ionizing radiation

    International Nuclear Information System (INIS)

    Silva, Josenilda Maria da; Silva, Juliana Pizarro; Spoto, Marta Helena Fillet

    2007-01-01

    The physiological and enzymatic post-harvest characteristics of the pineapple cultivar Smooth Cayenne were evaluated after the fruits were gamma-irradiated with doses of 100 and 150 Gy and the fruits were stored for 10, 20 and 30 days at 12 deg C (±1) and relative humidity of 85% (±5). Physiological and enzymatic analyses were made for each storage period to evaluate the alterations resulting from the application of ionizing radiation. Control specimens showed higher values of soluble pectins, total pectins, reducing sugars, sucrose and total sugars and lower values of polyphenyloxidase and polygalacturonase enzyme activities. All the analyses indicated that storage time is a significantly influencing factor. The 100 Gy dosage and 20-day storage period presented the best results from the standpoint of maturation and conservation of the fruits quality. (author)

  15. Improving biogas production from microalgae by enzymatic pretreatment.

    Science.gov (United States)

    Passos, Fabiana; Hom-Diaz, Andrea; Blanquez, Paqui; Vicent, Teresa; Ferrer, Ivet

    2016-01-01

    In this study, enzymatic pretreatment of microalgal biomass was investigated under different conditions and evaluated using biochemical methane potential (BMP) tests. Cellulase, glucohydrolase and an enzyme mix composed of cellulase, glucohydrolase and xylanase were selected based on the microalgae cell wall composition (cellulose, hemicellulose, pectin and glycoprotein). All of them increased organic matter solubilisation, obtaining high values already after 6h of pretreatment with an enzyme dose of 1% for cellulase and the enzyme mix. BMP tests with pretreated microalgae showed a methane yield increase of 8 and 15% for cellulase and the enzyme mix, respectively. Prospective research should evaluate enzymatic pretreatments in continuous anaerobic reactors so as to estimate the energy balance and economic cost of the process. Copyright © 2015 Elsevier Ltd. All rights reserved.

  16. Role of enzymatic and non enzymatic antioxidant in ameliorating salinity induced damage in nostoc muscorum

    International Nuclear Information System (INIS)

    Hend, A.; Abeer, A.; Allah, A.

    2015-01-01

    Presence of high salt concentration in the growth medium adversely affected the plant growth and productivity by altering its metabolic activities. Experiments were conducted on cyanobacteriaum Nostoc muscorum grown in nitrogen free medium supplemented with 250 mM NaCl to evaluate the salt stress induced changes in growth, antioxidants and lipid composition. Salt stress significantly reduced the growth and physio-biochemical attributes. Salt stress increased malonaldehyde content thereby causing alterations in the lipid fraction. Significant reduction in polyunsaturated fatty acids including phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylglycerol (PG), phosphatidylinositol (PI) and phosphatidylserine (PS) was observed. Where as diacylglycerol, sterol ester and non-esterified fatty acids were increased. Activities of antioxidant enzymes and contents of non-enzymatic antioxidants including glutathione enhanced due to salt stress. An increase in accumulation of proline was also observed. Hence increased activity of antioxidants and altered fatty acid composition was observed in salt stressed Nostoc muscorum. (author)

  17. Thermal and enzymatic recovering of proteins from untanned leather waste.

    Science.gov (United States)

    Bajza, Z; Vrucek, V

    2001-01-01

    The laboratory trials of a process to treat untanned leather waste to isolate valuable protein products are presented. In this comparative study, both thermal and enzymatic treatments of leather waste were performed. The enzymatic method utilizes commercially available alkaline protease at moderate temperatures and for short periods of time. The concentration of the enzyme was 500 units per gram of leather waste which makes the method cost-effective. Amino acid composition in the hydrolysate obtained by the enzyme hydrolysis of untanned leather waste is determined. Chemical and physical properties of protein powder products from untanned leather waste were evaluated by spectrophotometric and chromatographic methods and by use of electron microscope. The results of microbiological assays confirm that these products agree to food safety standards. This relatively simple treatment of untanned leather waste may provide a practical and economical solution to the disposal of potentially dangerous waste.

  18. Detection of extracellular enzymatic activity in microorganisms ...

    African Journals Online (AJOL)

    sunny t

    2015-09-18

    Sep 18, 2015 ... microorganisms with all three enzymatic activities, thereby establishing these techniques as ... supplemented at 1% with vegetable oils, including olive (OLI) ..... cepacia lipase for biodiesel fuel production from soybean oil.

  19. Electrochemical, Chemical and Enzymatic Oxidations of Phenothiazines

    NARCIS (Netherlands)

    Blankert, B.; Hayen, H.; van Leeuwen, S.M.; Karst, U.; Bodoki, E.; Lotrean, S.; Sandulescu, R.; Mora Diaz, N.; Dominguez, O.; Arcos, J.; Kauffmann, J.-M.

    2005-01-01

    The oxidation of several phenothiazine drugs (phenothiazine, promethazine hydrochloride, promazine hydrochloride, trimeprazine hydrochloride and ethopropazine hydrochloride) has been carried out in aqueous acidic media by electrochemical, chemical and enzymatic methods. The chemical oxidation was

  20. Detection of extracellular enzymatic activity in microorganisms ...

    African Journals Online (AJOL)

    Detection of extracellular enzymatic activity in microorganisms isolated from waste vegetable oil contaminated soil using plate methodologies. Eugenia G. Ortiz Lechuga, Isela Quintero Zapata, Katiushka Arévalo Niño ...

  1. Enzymatic hydrolysis of plant extracts containing inulin

    Energy Technology Data Exchange (ETDEWEB)

    Guiraud, J.P.; Galzy, P.

    1981-10-01

    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 3.2.1.7). 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).

  2. Kinetics of enzymatic hydrolysis of methyl ricinoleate

    OpenAIRE

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

    2015-01-01

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

  3. Antimicrobial and enzymatic activity of anemophilous fungi of a public university in Brazil

    Directory of Open Access Journals (Sweden)

    LAUREANA V. SOBRAL

    2017-10-01

    Full Text Available ABSTRACT To the fungal microbiota the UFPE and biotechnological potential enzymatic and antimicrobial production. Air conditioned environments were sampled using a passive sedimentation technique, the air I ratio and the presence of aflatoxigenic strains evaluated for ANVISA. Icelles were to determine the enzymatic activity of lipase, amylase and protease metabolic liquids to determine antimicrobial activity. Diversity was observed in all CAV environments, CFU/m3 ranged from 14 to 290 and I/E ratio from 0.1 to 1.5. The of the fungal genera were: Aspergillus (50%, Penicillium (21%, Talaromyces (14%, Curvularia and Paecilomyces (7% each. Aspergillus sydowii (Bainier & Sartory Thom & Church presented enzymatic activity and the Talaromyces purpureogenus Samson, Yilmaz, Houbraken, Spierenb., Seifert, Peterson, Varga & Frisvad presented antibacterial activity against all bacteria that all environments present fungal species biodiversity no toxigenic or pathogenic fungi were found, according to ANVISA legislation for conditioned environments and airborne filamentous fungi present potential for enzymatic and antimicrobial activity.

  4. Application of enzymatic methods for chia (Salvia hispanica L oil extraction

    Directory of Open Access Journals (Sweden)

    Norma Ciau-Solís

    2016-07-01

    Full Text Available Aim. The aim was to evaluate the use of different enzymatic treatments on the oil extraction yield from Chia (Salvia hispanica L. seeds Methods. Enzymatic extraction was performed by treating of whole and degummed chia flours with different conditions of enzyme concentration, pH and temperature. Commercial enzymes were employed: Viscozyme LTM (endo-1,3 (4-betaglucanase derived from Aspergillus aculeatus, with 100 FBG g (Beta Glucanase-unit Fungal and Neutrase0.8LTM, neutral protease with 0.8 AU-NH/g of activity, derived from Bacillus amyloliquefaciens. Results. All treatments of enzymatic oil extraction were different (P <0.05 and the maximum oil yield obtained was 9.35%. Conclusion. Oil extraction using enzymatic methods is not a viable for chia seed

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

    Directory of Open Access Journals (Sweden)

    Raquel Cristine Kuhn

    2015-05-01

    Full Text Available 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 period were measured. The best results for the ultrasound-assisted enzymatic hydrolysis was obtained using 3 wt% of enzyme, 60 oC and moisture content of 65 wt%, yielding 0.38 g sugar/g rice bran, whereas for the hydrolysis in the absence of ultrasound the highest yield was 0.20 g sugar/g rice bran using 3 wt% of enzyme, 60 oC and moisture content of 50 wt%. The use of ultrasound-assisted enzymatic hydrolysis of rice bran was intensified, obtaining around 74% more fermentable sugar than in the absence, showing that the use of ultrasound is a promising technology to be used in enzymatic reaction as an alternative of process intensification.

  6. Enzymatic approaches to rare sugar production.

    Science.gov (United States)

    Zhang, Wenli; Zhang, Tao; Jiang, Bo; Mu, Wanmeng

    Rare sugars have recently attracted much attention because of their potential applications in the food, nutraceutical, and pharmaceutical industries. A systematic strategy for enzymatic production of rare sugars, named Izumoring, was developed >10years ago. The strategy consists of aldose-ketose isomerization, ketose C-3 epimerization, and monosaccharide oxidation-reduction. Recent development of the Izumoring strategy is reviewed herein, especially the genetic approaches to the improvement of rare sugar-producing enzymes and the applications of target-oriented bioconversion. In addition, novel non-Izumoring enzymatic approaches are also summarized, including enzymatic condensation, phosphorylation-dephosphorylation cascade reaction, aldose epimerization, ulosonic acid decarboxylation, and biosynthesis of rare disaccharides. Copyright © 2017 Elsevier Inc. All rights reserved.

  7. Enzymatic Inverse Opal Hydrogel Particles for Biocatalyst.

    Science.gov (United States)

    Wang, Huan; Gu, Hongcheng; Chen, Zhuoyue; Shang, Luoran; Zhao, Ze; Gu, Zhongze; Zhao, Yuanjin

    2017-04-19

    Enzymatic carriers have a demonstrated value for chemical reactions and industrial applications. Here, we present a novel kind of inverse opal hydrogel particles as the enzymatic carriers. The particles were negatively replicated from spherical colloidal crystal templates by using magnetic nanoparticles tagged acrylamide hydrogel. Thus, they were endowed with the features of monodispersity, small volume, complete penetrating structure, and controllable motion, which are all beneficial for improving the efficiency of biocatalysis. In addition, due to the ordered porous nanostructure, the inverse opal hydrogel particles were imparted with unique photonic band gaps (PBGs) and vivid structural colors for encoding varieties of immobilized enzymes and for constructing a multienzymes biocatalysis system. These features of the inverse opal hydrogel particles indicate that they are ideal enzymatic carriers for biocatalysis.

  8. Enzymatic biodiesel production: Technical and economical considerations

    DEFF Research Database (Denmark)

    Munk Nielsen, Per; Brask, Jesper; Fjerbæk, Lene

    2008-01-01

    It is well documented in the literature that enzymatic processing of oils and fats for biodiesel is technically feasible. However, with very few exceptions, enzyme technology is not currently used in commercial-scale biodiesel production. This is mainly due to non-optimized process design...... and a lack of available costeffective enzymes. The technology to re-use enzymes has typically proven insufficient for the processes to be competitive. However, literature data documenting the productivity of enzymatic biodiesel together with the development of new immobilization technology indicates...... that enzyme catalysts can become cost effective compared to chemical processing. This work reviews the enzymatic processing of oils and fats into biodiesel with focus on process design and economy....

  9. Operation and Control of Enzymatic Biodiesel Production

    DEFF Research Database (Denmark)

    Price, Jason Anthony; Huusom, Jakob Kjøbsted; Nordblad, Mathias

    This work explores the control of biodiesel production via an enzymatic catalyst. The process involves the transesterification of oils/fats with an alcohol (usually methanol or ethanol), using enzymatic catalysts to generate mono-alkyl esters (the basis of biodiesel) and glycerol as by......-product. Current literature indicates that enzymatic processing of oils and fats to produce biodiesel is technically feasible and developments in immobilization technology indicate that enzyme catalysts can become cost effective compared to chemical processing. However, with very few exceptions, enzyme technology...... is not currently used in commercial-scale biodiesel production. This is mainly due to non-optimized process designs, which do not use the full potential of the catalysts in a cost-efficient way. Furthermore is it unclear what process variables need to be monitored and controlled to ensure optimal economics...

  10. Fed-Batch Feeding Strategies for Enzymatic Biodiesel Production

    DEFF Research Database (Denmark)

    Price, Jason Anthony; Nordblad, Mathias; Woodley, John

    2014-01-01

    of the differences in the interfacial and bulk concentrations of the enzyme. The model is then used to evaluate various feeding strategies to improve the enzymatic biodiesel production. The feeding strategies investigated, gave insight into how the methanol should be fed to potentially mitigate enzyme deactivation...... while improving the biodiesel yield. The best experimental results gave a yield of 703 .76 g FAME L-1 and a reactor productivity of 28.12 g FAME L-1 h-1. In comparison, to reach the same yield, the optimised two step feeding strategy took 6.25 hours less, which equates to an increase the reactor...

  11. Tandem and sequential multi-enzymatic syntheses

    NARCIS (Netherlands)

    Kim, B.G.; Ahn, J.H.; Sello, G.; Di Gennaro, P.; van Herk, T.; Hartog, A.F.; Wever, R.; Oroz-Guinea, I.; Sánchez-Moreno, I.; García-Junceda, E.; Wu, B.; Szymanski, W.; Feringa, B.L.; Janssen, D.B.; Villo, L.; Kreen, M.; Kudryashova, M.; Metsala, A.; Tamp, S.; Lille, ü.; Pehk, T.; Parve, O.; McClean, K.; Eddowes, P.; Whittall, J.; Sutton, P.W.

    2012-01-01

    This chapter contains sections titled: Production of Isorhamnetin 3-O-Glucoside in Escherichia coli Using Engineered Glycosyltransferase Multienzymatic Preparation of (−)-3-(Oxiran-2-yl)Benzoic Acid Enzymatic Synthesis of Carbohydrates from Dihydroxyacetone and Aldehydes by a One Pot Enzyme Cascade

  12. Enzymatic assay for methotrexate in erythrocytes

    DEFF Research Database (Denmark)

    Schrøder, H; Heinsvig, E M

    1985-01-01

    Methotrexate (MTX) accumulates in erythrocytes in MTX-treated patients. We present a modified enzymatic assay measuring MTX concentrations between 10 and 60 nmol/l in erythrocytes, adapted for a centrifugal analyser (Cobas Bio). About 40 patient's samples could be analysed within 1 h. The detection...

  13. Enzymatic hydrolysis of pretreated soybean straw

    International Nuclear Information System (INIS)

    Xu Zhong; Wang Qunhui; Jiang Zhaohua; Yang Xuexin; Ji Yongzhen

    2007-01-01

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

  14. Starch: chemistry, microstructure, processing and enzymatic degradation

    Science.gov (United States)

    Starch is recognized as one of the most abundant and important commodities containing value added attributes for a vast number of industrial applications. Its chemistry, structure, property and susceptibility to various chemical, physical and enzymatic modifications offer a high technological value ...

  15. Coated tube for immunochemical and enzymatic assays

    International Nuclear Information System (INIS)

    Brown, J.L.; Lin, W.H.-T.; Woods, J.W.

    1979-01-01

    Containers such as test tubes suitable for use in solid phase immunochemical, enzymatical and particularly radioimmunoassay procedures are described. The lower part of the tube is a polymer, coated with an inert protein to which a biologically active substance eg an antibody to triiodothyronine, thyroxine or digoxin, is attached. (U.K.)

  16. Starch facilitates enzymatic wheat gluten hydrolysis

    NARCIS (Netherlands)

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

    2015-01-01

    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)

  17. Enzymatic conversion of lignocellulose into fermentable sugars

    DEFF Research Database (Denmark)

    Jørgensen, Henning; Kristensen, Jan Bach; Felby, Claus

    2007-01-01

    and hemicelluloses but these are not readily accessible to enzymatic hydrolysis and require a pretreatment, which causes an extensive modification of the lignocellulosic structure. A number of pretreatment technologies are under development and being tested in pilot scale. Hydrolysis of lignocellulose carbohydrates...

  18. Enzymatic production of polysaccharides from gum tragacanth

    DEFF Research Database (Denmark)

    2014-01-01

    Plant polysaccharides, relating to the field of natural probiotic components, can comprise structures similar to human milk oligosaccharides. A method for enzymatic hydrolysis of gum tragacanth from the bush-like legumes of the genus Astragalus, using a combination of pectin hydrolases...

  19. Algorithms Development in Detection of the Gelatinization Process during Enzymatic ‘Dodol’ Processing

    Directory of Open Access Journals (Sweden)

    Azman Hamzah

    2013-09-01

    Full Text Available Computer vision systems have found wide application in foods processing industry to perform quality evaluation. The systems enable to replace human inspectors for the evaluation of a variety of quality attributes. This paper describes the implementation of the Fast Fourier Transform and Kalman filtering algorithms to detect the glutinous rice flour slurry (GRFS gelatinization in an enzymatic „dodol. processing. The onset of the GRFS gelatinization is critical in determining the quality of an enzymatic „dodol.. Combinations of these two algorithms were able to detect the gelatinization of the GRFS. The result shows that the gelatinization of the GRFS was at the time range of 11.75 minutes to 14.75 minutes for 24 batches of processing. This paper will highlight the capability of computer vision using our proposed algorithms in monitoring and controlling of an enzymatic „dodol. processing via image processing technology.

  20. Algorithms Development in Detection of the Gelatinization Process during Enzymatic ‘Dodol’ Processing

    Directory of Open Access Journals (Sweden)

    Azman Hamzah

    2007-11-01

    Full Text Available Computer vision systems have found wide application in foods processing industry to perform the quality evaluation. The systems enable to replace human inspectors for the evaluation of a variety of quality attributes. This paper describes the implementation of the Fast Fourier Transform and Kalman filtering algorithms to detect the glutinous rice flour slurry (GRFS gelatinization in an enzymatic ‘dodol’ processing. The onset of the GRFS gelatinization is critical in determining the quality of an enzymatic ‘dodol’. Combinations of these two algorithms were able to detect the gelatinization of the GRFS. The result shows that the gelatinization of the GRFS was at the time range of 11.75 minutes to 15.33 minutes for 20 batches of processing. This paper will highlight the capability of computer vision using our proposed algorithms in monitoring and controlling of an enzymatic ‘dodol’ processing via image processing technology.

  1. Effect of aflatoxin B1 on growth and enzymatic activity of a native strain of Bacillus sp

    Directory of Open Access Journals (Sweden)

    Alex Sáez Vega

    2004-01-01

    Full Text Available The effect of different aflatoxin B1 (AFAB1 concentrations on alkaline protease growth and enzymatic activity was evaluated; a native strain of alkalophilic Bacillus sp cultivated in CSL (Corn Steep Liquor was used. It was found that the effect of AFAB1 on the strain inhibited its growth and enzymatic activity to 1 ppm, showing that the strain is highly sensible to AFAB1, meaning that medium obtained f rom Colombian corn contaminated with this mycotoxin cannot be easily used. Concentrations less than 0.1 ppm did not affect growth and enzymatic activity. Key words: Bacillus, aflatoxin, alkaline proteases.

  2. Enzymatic hydrolysis of pretreated barley and wheat straw

    DEFF Research Database (Denmark)

    Rosgaard, Lisa

    2007-01-01

    . The work involved evaluation of 1) possible ways to increase the glucose release from the commercial cellulase product Celluclast by boosting with other enzyme activities to increase the enzymatic hydrolysis, 2) comparing differently pretreated feedstock substrates and 3) evaluating a fed-batch substrate...... mixture resulted in a glucose release corresponding to ~84 % of the glucose release from Celluclast. It was therefore suggested that other enzyme activities than the 4 four main cellulase activities in Celluclast are necessary for optimal hydrolysis of lignocellulose. Even though Celluclast...... is a multicomponent cellulase mixture, there are still possibilities for further improvement in terms of providing the most efficient cellulase mixture for lignocellulose hydrolysis. It was shown that substrates evaluated all had some residual hemicellulose in the solid cellulose fraction after pretreatment...

  3. Understanding of alkaline pretreatment parameters for corn stover enzymatic saccharification

    Directory of Open Access Journals (Sweden)

    Chen Ye

    2013-01-01

    Full Text Available Abstract Background Previous research on alkaline pretreatment has mainly focused on optimization of the process parameters to improve substrate digestibility. To achieve satisfactory sugar yield, extremely high chemical loading and enzyme dosages were typically used. Relatively little attention has been paid to reduction of chemical consumption and process waste management, which has proven to be an indispensable component of the bio-refineries. To indicate alkali strength, both alkali concentration in pretreatment solution (g alkali/g pretreatment liquor or g alkali/L pretreatment liquor and alkali loading based on biomass solids (g alkali/g dry biomass have been widely used. The dual approaches make it difficult to compare the chemical consumption in different process scenarios while evaluating the cost effectiveness of this pretreatment technology. The current work addresses these issues through pretreatment of corn stover at various combinations of pretreatment conditions. Enzymatic hydrolysis with different enzyme blends was subsequently performed to identify the effects of pretreatment parameters on substrate digestibility as well as process operational and capital costs. Results The results showed that sodium hydroxide loading is the most dominant variable for enzymatic digestibility. To reach 70% glucan conversion while avoiding extensive degradation of hemicellulose, approximately 0.08 g NaOH/g corn stover was required. It was also concluded that alkali loading based on total solids (g NaOH/g dry biomass governs the pretreatment efficiency. Supplementing cellulase with accessory enzymes such as α-arabinofuranosidase and β-xylosidase significantly improved the conversion of the hemicellulose by 6–17%. Conclusions The current work presents the impact of alkaline pretreatment parameters on the enzymatic hydrolysis of corn stover as well as the process operational and capital investment costs. The high chemical consumption for alkaline

  4. Mechano-Enzymatic Deconstruction with a New Enzymatic Cocktail to Enhance Enzymatic Hydrolysis and Bioethanol Fermentation of Two Macroalgae Species

    Directory of Open Access Journals (Sweden)

    Sameh Amamou

    2018-01-01

    Full Text Available The aim of this study was to explore the efficiency of a mechano-enzymatic deconstruction of two macroalgae species for sugars and bioethanol production, by using a new enzymatic cocktail (Haliatase and two types of milling modes (vibro-ball: VBM and centrifugal milling: CM. By increasing the enzymatic concentration from 3.4 to 30 g/L, the total sugars released after 72 h of hydrolysis increased (from 6.7 to 13.1 g/100 g TS and from 7.95 to 10.8 g/100 g TS for the green algae U. lactuca and the red algae G. sesquipedale, respectively. Conversely, total sugars released from G. sesquipedale increased (up to 126% and 129% after VBM and CM, respectively. The best bioethanol yield (6 geth/100 g TS was reached after 72 h of fermentation of U. lactuca and no increase was obtained after centrifugal milling. The latter led to an enhancement of the ethanol yield of G. sesquipedale (from 2 to 4 g/100 g TS.

  5. Enzymatic degradation of polycaprolactone–gelatin blend

    International Nuclear Information System (INIS)

    Banerjee, Aditi; Chatterjee, Kaushik; Madras, Giridhar

    2015-01-01

    Blends of polycaprolactone (PCL), a synthetic polymer and gelatin, natural polymer offer a optimal combination of strength, water wettability and cytocompatibility for use as a resorbable biomaterial. The enzymatic degradation of PCL, gelatin and PCL–gelatin blended films was studied in the presence of lipase (Novozym 435, immobilized) and lysozyme. Novozym 435 degraded the PCL films whereas lysozyme degraded the gelatin. Though Novozym 435 and lysozyme individually could degrade PCL–gelatin blended films, the combination of these enzymes showed the highest degradation of these blended films. Moreover, the enzymatic degradation was much faster when fresh enzymes were added at regular intervals. The changes in physico-chemical properties of polymer films due to degradation were studied by scanning electron microscopy, Fourier transform infrared spectroscopy and differential scanning calorimetry. These results have important implications for designing resorbable biomedical implants. (paper)

  6. A singular enzymatic megacomplex from Bacillus subtilis.

    Science.gov (United States)

    Straight, Paul D; Fischbach, Michael A; Walsh, Christopher T; Rudner, David Z; Kolter, Roberto

    2007-01-02

    Nonribosomal peptide synthetases (NRPS), polyketide synthases (PKS), and hybrid NRPS/PKS are of particular interest, because they produce numerous therapeutic agents, have great potential for engineering novel compounds, and are the largest enzymes known. The predicted masses of known enzymatic assembly lines can reach almost 5 megadaltons, dwarfing even the ribosome (approximately 2.6 megadaltons). Despite their uniqueness and importance, little is known about the organization of these enzymes within the native producer cells. Here we report that an 80-kb gene cluster, which occupies approximately 2% of the Bacillus subtilis genome, encodes the subunits of approximately 2.5 megadalton active hybrid NRPS/PKS. Many copies of the NRPS/PKS assemble into a single organelle-like membrane-associated complex of tens to hundreds of megadaltons. Such an enzymatic megacomplex is unprecedented in bacterial subcellular organization and has important implications for engineering novel NRPS/PKSs.

  7. A Networks Approach to Modeling Enzymatic Reactions.

    Science.gov (United States)

    Imhof, P

    2016-01-01

    Modeling enzymatic reactions is a demanding task due to the complexity of the system, the many degrees of freedom involved and the complex, chemical, and conformational transitions associated with the reaction. Consequently, enzymatic reactions are not determined by precisely one reaction pathway. Hence, it is beneficial to obtain a comprehensive picture of possible reaction paths and competing mechanisms. By combining individually generated intermediate states and chemical transition steps a network of such pathways can be constructed. Transition networks are a discretized representation of a potential energy landscape consisting of a multitude of reaction pathways connecting the end states of the reaction. The graph structure of the network allows an easy identification of the energetically most favorable pathways as well as a number of alternative routes. © 2016 Elsevier Inc. All rights reserved.

  8. Production of MAG via enzymatic glycerolysis

    Science.gov (United States)

    Jamlus, Norul Naziraa Ahmad; Derawi, Darfizzi; Salimon, Jumat

    2015-09-01

    Enzymatic glycerolysis of a medium chain methyl ester, methyl laurate was performed using lipase Candida antarctica (Novozyme 435) for 6 hours at 55°C. The percentage of components mixture of product were determined by using gas chromatography technique. The enzymatic reaction was successfully produced monolaurin (45.9 %), dilaurin (47.1 %) and trilaurin (7.0 %) respectively. Thin layer chromatography (TLC) plate also showed a good separation of component spots. Fourier transformation infra-red (FTIR) spectrum showed the presence of ester carbonyl at wavenumber 1739.99 cm-1 and hydrogen bonded O-H at 3512.03 cm-1. The product is potentially to be used as emulsifier and additive in food industry, pharmaceutical, as well as antibacterial.

  9. Heavy metal pollution and soil enzymatic activity

    Energy Technology Data Exchange (ETDEWEB)

    Tyler, G

    1974-01-01

    The activity of hydrolytic soil enzymes was studied on spruce mor, polluted with Cu and Zn from a brass foundry in Sweden. Approximately straight regression lines were obtained between enzymatic activity or respiration rate and log Cu + Zn concentration, with highly significant negative regression coefficients for urease and acid phosphatase activity as well as respiration rate, whereas US -glucosidase activity was not measurably lower at high concentrations of Cu + Zn. 17 references, 5 figures.

  10. Enzymatic Activity Detection via Electrochemistry for Enceladus

    Science.gov (United States)

    Studemeister, Lucy; Koehne, Jessica; Quinn, Richard

    2017-01-01

    Electrochemical detection of biological molecules is a pertinent topic and application in many fields such as medicine, environmental spills, and life detection in space. Proteases, a class of molecules of interest in the search for life, catalyze the hydrolysis of peptides. Trypsin, a specific protease, was chosen to investigate an optimized enzyme detection system using electrochemistry. This study aims at providing the ideal functionalization of an electrode that can reliably detect a signal indicative of an enzymatic reaction from an Enceladus sample.

  11. Enzymatic Synthesis of Biobased Polyesters and Polyamides

    Directory of Open Access Journals (Sweden)

    Yi Jiang

    2016-06-01

    Full Text Available Nowadays, “green” is a hot topic almost everywhere, from retailers to universities to industries; and achieving a green status has become a universal aim. However, polymers are commonly considered not to be “green”, being associated with massive energy consumption and severe pollution problems (for example, the “Plastic Soup” as a public stereotype. To achieve green polymers, three elements should be entailed: (1 green raw materials, catalysts and solvents; (2 eco-friendly synthesis processes; and (3 sustainable polymers with a low carbon footprint, for example, (biodegradable polymers or polymers which can be recycled or disposed with a gentle environmental impact. By utilizing biobased monomers in enzymatic polymerizations, many advantageous green aspects can be fulfilled. For example, biobased monomers and enzyme catalysts are renewable materials that are derived from biomass feedstocks; enzymatic polymerizations are clean and energy saving processes; and no toxic residuals contaminate the final products. Therefore, synthesis of renewable polymers via enzymatic polymerizations of biobased monomers provides an opportunity for achieving green polymers and a future sustainable polymer industry, which will eventually play an essential role for realizing and maintaining a biobased and sustainable society.

  12. Enzymatic transformation of nonfood biomass to starch

    Science.gov (United States)

    You, Chun; Chen, Hongge; Myung, Suwan; Sathitsuksanoh, Noppadon; Ma, Hui; Zhang, Xiao-Zhou; Li, Jianyong; Zhang, Y.-H. Percival

    2013-01-01

    The global demand for food could double in another 40 y owing to growth in the population and food consumption per capita. To meet the world’s future food and sustainability needs for biofuels and renewable materials, the production of starch-rich cereals and cellulose-rich bioenergy plants must grow substantially while minimizing agriculture’s environmental footprint and conserving biodiversity. Here we demonstrate one-pot enzymatic conversion of pretreated biomass to starch through a nonnatural synthetic enzymatic pathway composed of endoglucanase, cellobiohydrolyase, cellobiose phosphorylase, and alpha-glucan phosphorylase originating from bacterial, fungal, and plant sources. A special polypeptide cap in potato alpha-glucan phosphorylase was essential to push a partially hydrolyzed intermediate of cellulose forward to the synthesis of amylose. Up to 30% of the anhydroglucose units in cellulose were converted to starch; the remaining cellulose was hydrolyzed to glucose suitable for ethanol production by yeast in the same bioreactor. Next-generation biorefineries based on simultaneous enzymatic biotransformation and microbial fermentation could address the food, biofuels, and environment trilemma. PMID:23589840

  13. Microbial Enzymatic Degradation of Biodegradable Plastics.

    Science.gov (United States)

    Roohi; Bano, Kulsoom; Kuddus, Mohammed; Zaheer, Mohammed R; Zia, Qamar; Khan, Mohammed F; Ashraf, Ghulam Md; Gupta, Anamika; Aliev, Gjumrakch

    2017-01-01

    The renewable feedstock derived biodegradable plastics are important in various industries such as packaging, agricultural, paper coating, garbage bags and biomedical implants. The increasing water and waste pollution due to the available decomposition methods of plastic degradation have led to the emergence of biodegradable plastics and biological degradation with microbial (bacteria and fungi) extracellular enzymes. The microbes utilize biodegradable polymers as the substrate under starvation and in unavailability of microbial nutrients. Microbial enzymatic degradation is suitable from bioremediation point of view as no waste accumulation occurs. It is important to understand the microbial interaction and mechanism involved in the enzymatic degradation of biodegradable plastics under the influence of several environmental factors such as applied pH, thermo-stability, substrate molecular weight and/or complexity. To study the surface erosion of polymer film is another approach for hydrolytic degradation characteristion. The degradation of biopolymer is associated with the production of low molecular weight monomer and generation of carbon dioxide, methane and water molecule. This review reported the degradation study of various existing biodegradable plastics along with the potent degrading microbes (bacteria and fungi). Patents available on plastic biodegradation with biotechnological significance is also summarized in this paper. This paper assesses that new disposal technique should be adopted for the degradation of polymers and further research is required for the economical production of biodegradable plastics along with their enzymatic degradation. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  14. Impact of the fouling mechanism on enzymatic depolymerization of xylan in different configurations of membrane reactors

    DEFF Research Database (Denmark)

    Mohd Sueb, Mohd Shafiq Bin; Luo, Jianquan; Meyer, Anne S.

    2017-01-01

    In order to maximize enzymatic xylan depolymerization while simultaneously purifying the resulting monosaccharide (xylose), different ultrafiltration (UF) membrane reactor configurations were evaluated. Initial results showed that the two hydrolytic enzymes required for complete depolymerization...... which hindered enzymatic attack in addition to fouling. Reaction with both enzymes followed by UF was found to be the optimal configuration, providing at least 40% higher xylan hydrolysis than the cascade configuration (involving sequential reaction with each of the enzymes separately......) and the simultaneous reaction-filtration with both enzymes, respectively. This study thus confirmed that the reactor configuration has a crucial impact on the performance of both the reaction and the separation process of xylose during enzymatic xylan degradation, and that the type of fouling mechanism varies...

  15. Production of xylitol from corn cob hydrolysate through acid and enzymatic hydrolysis by yeast

    Science.gov (United States)

    Mardawati, Efri; Andoyo, R.; Syukra, K. A.; Kresnowati, MTAP; Bindar, Y.

    2018-03-01

    The abundance of corn production in Indonesia offers the potential for its application as the raw material for biorefinery process. The hemicellulose content in corn cobs can be considered to be used as a raw material for xylitol production. The purpose of this research was to study the effect of hydrolysis methods for xylitol production and the effect of the hydrolyzed corn cobs to produce xylitol through fermentation. Hydrolysis methods that would be evaluated were acid and enzymatic hydrolysis. The result showed that the xylitol yield of fermented solution using enzymatic hydrolysates was 0.216 g-xylitol/g-xylose, which was higher than the one that used acid hydrolysates, which was 0.100 g-xylitol/g-xylose. Moreover, the specific growth rate of biomass in fermentation using enzymatic hydrolysates was also higher than the one that used acid hydrolysates, 0.039/h compared to 0.0056/h.

  16. Comparison of Enzymatic and Ultrasonic Extraction of Albumin from Defatted Pumpkin (Cucurbita pepo Seed Powder

    Directory of Open Access Journals (Sweden)

    Gia Loi Tu

    2015-01-01

    Full Text Available In this study, ultrasound- and enzyme-assisted extractions of albumin (water-soluble protein group from defatted pumpkin (Cucurbita pepo seed powder were compared. Both advanced extraction techniques strongly increased the albumin yield in comparison with conventional extraction. The extraction rate was two times faster in the ultrasonic extraction than in the enzymatic extraction. However, the maximum albumin yield was 16 % higher when using enzymatic extraction. Functional properties of the pumpkin seed albumin concentrates obtained using the enzymatic, ultrasonic and conventional methods were then evaluated. Use of hydrolase for degradation of cell wall of the plant material did not change the functional properties of the albumin concentrate in comparison with the conventional extraction. The ultrasonic extraction enhanced water-holding, oil-holding and emulsifying capacities of the pumpkin seed albumin concentrate, but slightly reduced the foaming capacity, and emulsion and foam stability.

  17. Comparison of different pretreatment strategies for enzymatic hydrolysis of wheat and barley straw

    DEFF Research Database (Denmark)

    Rosgaard, Lisa; Pedersen, Sven; Meyer, Anne Boye Strunge

    2007-01-01

    In biomass-to-ethanol processes a physico-chemical pretreatment of the lignocellulosic biomass is a critical requirement for enhancing the accessibility of the cellulose substrate to enzymatic attack. This report evaluates the efficacy on barley and wheat straw of three different pretreatment pro...

  18. Enzymatic polymerization of bio-based monomers for applications in hydrogels and coatings

    DEFF Research Database (Denmark)

    Hoffmann, Christian; Nguyen, Hiep Dinh; Storgaard, Thomas

    of the enzymatic catalysts that can provide control over polymer structure in functional polymers. Lipase catalyzed polymerizations (specifically CALB) has been applied to prepare functional polyesters and to evaluate the possibilities of using less stable bio-based monomers such as itaconic acid or its...

  19. Optimization of Pretreatment and Enzymatic Saccharification of Cogon Grass Prior Ethanol Production

    OpenAIRE

    Jhalique Jane R. Fojas; Ernesto J. Del Rosario

    2013-01-01

    The dilute acid pretreatment and enzymatic saccharification of lignocellulosic substrate, cogon grass (Imperata cylindrical, L.) was optimized prior ethanol fermentation using simultaneous saccharification and fermentation (SSF) method. The optimum pretreatment conditions, temperature, sulfuric acid concentration, and reaction time were evaluated by determining the maximum sugar yield at constant enzyme loading. Cogon grass, at 10% w/v substrate loading, has optimum pretr...

  20. Improvement of Soybean Oil Solvent Extraction through Enzymatic Pretreatment

    Directory of Open Access Journals (Sweden)

    F. V. Grasso

    2012-01-01

    Full Text Available The purpose of this study is to evaluate multienzyme hydrolysis as a pretreatment option to improve soybean oil solvent extraction and its eventual adaptation to conventional processes. Enzymatic action causes the degradation of the cell structures that contain oil. Improvements in terms of extraction, yield, and extraction rate are expected to be achieved. Soybean flakes and collets were used as materials and hexane was used as a solvent. Temperature, pH, and incubation time were optimized and diffusion coefficients were estimated for each solid. Extractions were carried out in a column, oil content was determined according to time, and a mathematical model was developed to describe the system. The optimum conditions obtained were pH 5.4, 38°C, and 9.7 h, and pH 5.8, 44°C, and 5.8h of treatment for flakes and collets, respectively. Hydrolyzed solids exhibited a higher yield. Diffusion coefficients were estimated between 10-11 and 10-10. The highest diffusion coefficient was obtained for hydrolyzed collets. 0.73 g oil/mL and 0.7 g oil/mL were obtained at 240 s in a column for collets and flakes, respectively. Hydrolyzed solids exhibited a higher yield. The enzymatic incubation accelerates the extraction rate and allows for higher yield. The proposed model proved to be appropriate.

  1. Enzymatic production of pectic oligosaccharides from onion skins.

    Science.gov (United States)

    Babbar, Neha; Baldassarre, Stefania; Maesen, Miranda; Prandi, Barbara; Dejonghe, Winnie; Sforza, Stefano; Elst, Kathy

    2016-08-01

    Onion skins are evaluated as a new raw material for the enzymatic production of pectic oligosaccharides (POS) with a targeted degree of polymerization (DP). The process is based on a two-stage process consisting of a chelator-based crude pectin extraction followed by a controlled enzymatic hydrolysis. Treatment of the extracted crude onion skin's pectin with various enzymes (EPG-M2, Viscozyme and Pectinase) shows that EPG-M2 is the most appropriate enzyme for tailored POS production. The experiments reveal that the highest amount of DP2 and DP3 is obtained at a time scale of 75-90min with an EPG-M2 concentration of 26IU/mL. At these conditions the production amounts 2.5-3.0% (w/w) d.m for DP2 and 5.5-5.6% (w/w) d.m for DP3 respectively. In contrast, maximum DP4 production of 5.2-5.5% (w/w) d.m. is obtained with 5.2IU/mL at a time scale of 15-30min. Detailed LC-MS analysis reveals the presence of more methylated oligomers compared to acetylated forms in the digests. Copyright © 2016 Elsevier Ltd. All rights reserved.

  2. Investigation of bi-enzymatic reactor based on hybrid monolith with nanoparticles embedded and its proteolytic characteristics.

    Science.gov (United States)

    Shangguan, Lulu; Zhang, Lingyi; Xiong, Zhichao; Ren, Jun; Zhang, Runsheng; Gao, Fangyuan; Zhang, Weibing

    2015-04-03

    The bottom-up strategy of proteomic profiling study based on mass spectrometer (MS) has drawn high attention. However, conventional solution-based digestion could not satisfy the demands of highly efficient and complete high throughput proteolysis of complex samples. We proposed a novel bi-enzymatic reactor by immobilizing two different enzymes (trypsin/chymotrypsin) onto a mixed support of hybrid organic-inorganic monolith with SBA-15 nanoparticles embedded. Typsin and chymotrypsin were crossly immobilized onto the mixed support by covalent bonding onto the monolith with glutaraldehyde as bridge reagent and chelation via copper ion onto the nanoparticles, respectively. Compared with single enzymatic reactors, the bi-enzymatic reactor improved the overall functional analysis of membrane proteins of rat liver by doubling the number of identified peptides (from 1184/1010 with trypsin/chymotrypsin enzymatic reactors to 2891 with bi-enzymatic reactor), which led to more proteins identified with deep coverage (from 452/336 to 620); the efficiency of the bi-enzymatic reactor is also better than that of solution-based tandem digestion, greatly shorting the digestion time from 24h to 50s. Moreover, more transmembrane proteins were identified by bi-enzymatic reactor (106) compared with solution-based tandem digestion (95) with the same two enzymes and enzymatic reactors with single enzyme immobilized (75 with trypsin and 66 with chymotrypsin). The proteolytic characteristics of the bi-enzymatic reactors were evaluated by applying them to digestion of rat liver proteins. The reactors showed good digestion capability for proteins with different hydrophobicity and molecular weight. Copyright © 2015 Elsevier B.V. All rights reserved.

  3. Enzymatic activity measured by microcalorimetry in soil amended with organic residues

    Directory of Open Access Journals (Sweden)

    Karina Cenciani

    2011-08-01

    Full Text Available Enzymatic activity is an important property for soil quality evaluation. Two sequences of experiments were carried out in order to evaluate the enzymatic activity in a soil (Rhodic Eutrudox amended with cattle manure, earthworm casts, or sewage sludges from the municipalities of Barueri and Franca. The activity of commercial enzymes was measured by microcalorimetry in the same soil samples after sterilization. In the first experiment, the enzyme activities of cellulase, protease, and urease were determined in the soil samples during a three month period. In the second sequence of experiments, the thermal effect of the commercial enzymes cellulase, protease, and urease on sterilized soil samples under the same tretaments was monitored for a period of 46 days. The experimental design was randomized and arranged as factorial scheme in five treatments x seven samplings with five replications. The treatment effects were statistically evaluated by one-way analysis of variance. Tukey´s test was used to compare means at p < 0.05. The presence of different sources of organic residues increased the enzymatic activity in the sampling period. Cattle manure induced the highest enzymatic activity, followed by municipal sewage sludge, whereas earthworm casts induced the lowest activity, but differed from control treatment. The thermal effect on the enzyme activity of commercial cellulase, protease, and urease showed a variety of time peaks. These values probably oscillated due to soil physical-chemical factors affecting the enzyme activity on the residues.

  4. Glucuronic Acid Derivatives in Enzymatic Biomass Degradation: Synthesis and Evaluation of Enzymatic Activity

    DEFF Research Database (Denmark)

    d'Errico, Clotilde

    An essential tool for biotechnology companies in enzyme development for biomass delignification is the access to well-defined model substrates. A deeper understanding of the enzymes substrate specificity can be used to address and optimize enzyme mixtures towards natural, complex substrates. Hence...

  5. Enzymatic Hydrolysis of Alkaline Pretreated Coconut Coir

    Directory of Open Access Journals (Sweden)

    Akbarningrum Fatmawati

    2013-06-01

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

  6. Enzymatic Hydrolysis of Pretreated Fibre Pressed Oil Palm Frond by using Sacchariseb C6

    Science.gov (United States)

    Hashim, F. S.; Yussof, H. W.; Zahari, M. A. K. M.; Rahman, R. A.; Illias, R. M.

    2017-06-01

    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 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.

  7. Enzymatic Hydrolysis of Oleuropein from Olea europea (Olive Leaf Extract and Antioxidant Activities

    Directory of Open Access Journals (Sweden)

    Jiao-Jiao Yuan

    2015-02-01

    Full Text Available Oleuropein (OE, the main polyphenol in olive leaf extract, is likely to decompose into hydroxytyrosol (HT and elenolic acid under the action of light, acid, base, high temperature. In the enzymatic process, the content of OE in olive leaf extract and enzyme are key factors that affect the yield of HT. A selective enzyme was screened from among 10 enzymes with a high OE degradation rate. A single factor (pH, temperature, time, enzyme quantity optimization process and a Box-Behnken design were studied for the enzymatic hydrolysis of 81.04% OE olive leaf extract. Additionally, enzymatic hydrolysis results with different substrates (38.6% and 81.04% OE were compared and the DPPH antioxidant properties were also evaluated. The result showed that the performance of hydrolysis treatments was best using hemicellulase as a bio-catalyst, and the high purity of OE in olive extract was beneficial to biotransform OE into HT. The optimal enzymatic conditions for achieving a maximal yield of HT content obtained by the regression were as follows: pH 5, temperature 55 °C and enzyme quantity 55 mg. The experimental result was 11.31% ± 0.15%, and the degradation rate of OE was 98.54%. From the present investigation of the antioxidant activity determined by the DPPH method, the phenol content and radical scavenging effect were both decreased after enzymatic hydrolysis by hemicellulase. However, a high antioxidant activity of the ethyl acetate extract enzymatic hydrolysate (IC50 = 41.82 μg/mL was demonstated. The results presented in this work suggested that hemicellulase has promising and attractive properties for industrial production of HT, and indicated that HT might be a valuable biological component for use in pharmaceutical products and functional foods.

  8. Isothermal calorimetry of enzymatic biodiesel reaction

    DEFF Research Database (Denmark)

    Fjerbæk Søtoft, Lene; Westh, Peter; Christensen, Knud Villy

    2010-01-01

      Isothermal calorimetry ITC has been used to investigate enzymatic biodiesel production. The transesterification of rapeseed oil with methanol and ethanol was catalyzed by the immobilized lipase Novozym 435 at 40°C. The ITC-experiments clearly demonstrate the possibilities of investigating complex...... and composition change in the system, the heat of reaction at 40°C for the two systems has been determined to -9.8 ± 0.9 kJ/mole biodiesel formed from rapeseed oil and methanol, and - 9.3 ± 0.7 kJ/mole when rapeseed oil and ethanol is used....

  9. Waste management and enzymatic treatment of Municipal Solid Waste

    DEFF Research Database (Denmark)

    Jensen, Jacob Wagner

    generation for subsequent biogas production. Municipal solid waste (MSW) is produced in large amounts every year in the developed part of the world. The household waste composition varies between geographical areas and between seasons. However the overall content of organic and degradable material is rather......The work carried out during the Ph.D. project is part of the Danish Energy Authority funded research project called PSO REnescience and is focussed on studying the enzymatic hydrolysis and liquefaction of waste biomass. The purpose of studying the liquefaction of waste biomass is uniform slurry...... constant between 50 - 60 % wet weight and therefore holds a potential for bioenergy production. The degradable fraction has positive effects for anaerobic digestion when evaluated to desired parameters of anaerobic digestion plants. Wanted parameters are: 1) high organic content (high volatile solid...

  10. A xylanase-aided enzymatic pretreatment facilitates cellulose nanofibrillation.

    Science.gov (United States)

    Long, Lingfeng; Tian, Dong; Hu, Jinguang; Wang, Fei; Saddler, Jack

    2017-11-01

    Although biological pretreatment of cellulosic fiber based on endoglucanases has shown some promise to facilitate cellulose nanofibrillation, its efficacy is still limited. In this study, a xylanase-aided endoglucanase pretreatment was assessed on the bleached hardwood and softwood Kraft pulps to facilitate the downstream cellulose nanofibrillation. Four commercial xylanase preparations were compared and the changes of major fiber physicochemical characteristics such as cellulose/hemicellulose content, gross fiber properties, fiber morphologies, cellulose accessibility/degree of polymerization (DP)/crystallinity were systematically evaluated before and after enzymatic pretreatment. It showed that the synergistic cooperation between endoglucanase and certain xylanase (Biobrite) could efficiently "open up" the hardwood Kraft pulp with limited carbohydrates degradation (cellulose nanofibrillation during mild sonication process (90Wh) with more uniform disintegrated nanofibril products (50-150nm, as assessed by scanning electron microscopy and UV-vis spectroscopy). Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. Enzymatic Activity of Candida spp. from Oral Cavity and Urine in Children with Nephrotic Syndrome.

    Science.gov (United States)

    Olczak-Kowalczyk, Dorota; Roszkowska-Blaim, Maria; Dąbkowska, Maria; Swoboda-Kopeć, Ewa; Gozdowski, Dariusz; Mizerska-Wasiak, Małgorzata; Demkow, Urszula; Pańczyk-Tomaszewska, Małgorzata

    2017-01-01

    Oral colonization with Candida spp. is not synonymous with a systemic active infection. The aim of the study was to evaluate enzymatic activity of Candida strains isolated from the oral cavity in patients with nephrotic syndrome (NS) and to compare it with the activity determined in urine. We studied 32 children with NS and 26 control healthy children. Children with NS were treated with glucocorticosteroids, cyclosporin A, mycophenolate mofetil or azathioprine. In all children, API-ZYM enzymatic tests were performed to evaluate hydrolytic enzymes of Candida isolated from the oral cavity and in urine. Candida spp. were isolated from the oral cavity in 11 patients with NS (34.4%), all receiving immunosuppressive treatment. All strains produced valine arylamidase, 9 alpha-glucosidase (E16), and 9 N-acetyl-beta-glucosaminidase (E18). A positive correlation between the presence of Candida in the oral cavity and E16 and E18 enzymatic activity in both oral cavity and urine was found. A dose of cyclosporin A had an effect on the enzymatic activity (p Candida invasion. The results of this study suggest that oral candida infection should be monitored in children with nephrotic syndrome, particularly those treated with immunosuppressive agents.

  12. Enzymatic deconstruction of xylan for biofuel production

    Science.gov (United States)

    DODD, DYLAN; CANN, ISAAC K. O.

    2010-01-01

    The combustion of fossil-derived fuels has a significant impact on atmospheric carbon dioxide (CO2) levels and correspondingly is an important contributor to anthropogenic global climate change. Plants have evolved photosynthetic mechanisms in which solar energy is used to fix CO2 into carbohydrates. Thus, combustion of biofuels, derived from plant biomass, can be considered a potentially carbon neutral process. One of the major limitations for efficient conversion of plant biomass to biofuels is the recalcitrant nature of the plant cell wall, which is composed mostly of lignocellulosic materials (lignin, cellulose, and hemicellulose). The heteropolymer xylan represents the most abundant hemicellulosic polysaccharide and is composed primarily of xylose, arabinose, and glucuronic acid. Microbes have evolved a plethora of enzymatic strategies for hydrolyzing xylan into its constituent sugars for subsequent fermentation to biofuels. Therefore, microorganisms are considered an important source of biocatalysts in the emerging biofuel industry. To produce an optimized enzymatic cocktail for xylan deconstruction, it will be valuable to gain insight at the molecular level of the chemical linkages and the mechanisms by which these enzymes recognize their substrates and catalyze their reactions. Recent advances in genomics, proteomics, and structural biology have revolutionized our understanding of the microbial xylanolytic enzymes. This review focuses on current understanding of the molecular basis for substrate specificity and catalysis by enzymes involved in xylan deconstruction. PMID:20431716

  13. Enzymatically crosslinked silk-hyaluronic acid hydrogels.

    Science.gov (United States)

    Raia, Nicole R; Partlow, Benjamin P; McGill, Meghan; Kimmerling, Erica Palma; Ghezzi, Chiara E; Kaplan, David L

    2017-07-01

    In this study, silk fibroin and hyaluronic acid (HA) were enzymatically crosslinked to form biocompatible composite hydrogels with tunable mechanical properties similar to that of native tissues. The formation of di-tyrosine crosslinks between silk fibroin proteins via horseradish peroxidase has resulted in a highly elastic hydrogel but exhibits time-dependent stiffening related to silk self-assembly and crystallization. Utilizing the same method of crosslinking, tyramine-substituted HA forms hydrophilic and bioactive hydrogels that tend to have limited mechanics and degrade rapidly. To address the limitations of these singular component scaffolds, HA was covalently crosslinked with silk, forming a composite hydrogel that exhibited both mechanical integrity and hydrophilicity. The composite hydrogels were assessed using unconfined compression and infrared spectroscopy to reveal of the physical properties over time in relation to polymer concentration. In addition, the hydrogels were characterized by enzymatic degradation and for cytotoxicity. Results showed that increasing HA concentration, decreased gelation time, increased degradation rate, and reduced changes that were observed over time in mechanics, water retention, and crystallization. These hydrogel composites provide a biologically relevant system with controllable temporal stiffening and elasticity, thus offering enhanced tunable scaffolds for short or long term applications in tissue engineering. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. Enzymatic activity of fungi isolated from crops

    Directory of Open Access Journals (Sweden)

    Wioletta A. Żukiewicz-Sobczak

    2016-12-01

    Full Text Available Aim: To detect and assess the activity of extracellular hydrolytic enzymes and to find differences in enzymograms between fungi isolated from wheat and rye samples and grown on Czapek-Dox Broth and Sabouraud Dextrose Broth enriched with cereal (wheat or rye. Isolated strains were also classified in the scale of biosafety levels (BSL. Material and methods: The study used 23 strains of fungi cultured from samples of wheat and rye (grain, grain dust obtained during threshing and soil collected in the Lublin region (eastern Poland. API ZYM test (bioMérieux was carried out according to the manufacturer’s instructions. Classification of BSL (Biosafety levels was based on the current literature. Results : High enzymatic activity was found in strains cultured in media containing 1% of wheat grain ( Bipolaris holmi, Penicillium decumbens and with an addition of 1% of rye grain ( Cladosporium herbarum, Aspergillus versicolor, Alternaria alternata . The total number of enzymes varied depending on the type of media, and in most cases it was higher in the culture where an addition of cereal grains was used. Conclusions : Isolated strains of fungi reveal differences in the profiles of the enzyme assay. It can be assumed that the substrate enriched in grains stimulate the higher activity of mold enzymes. Key words: enzymatic activity, mold fungi, zymogram, biohazards.

  15. Enzymatic hydrolysis of lactose of whey permeate

    Directory of Open Access Journals (Sweden)

    Karina Nascimento de Almeida

    2015-09-01

    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.

  16. Lactose hydrolysis in an enzymatic membrane reactor

    Energy Technology Data Exchange (ETDEWEB)

    Mertens, B; Huyghebaert, A

    1987-10-01

    The enzymatic hydrolysis of lactose in whey permeate with subsequent recuperation of Saccharomyces lactis lactase by means of ultrafiltration was investigated. In whey permeate, S. lactis lactase shows maximal activity at pH 6.5; the optimal temperature was found to be 45/sup 0/C and is limited by strong thermal inactivation beyond this temperature. High activity combined with acceptable thermal inactivation (< 10% after 5 h incubation) was established at 30/sup 0/C. S. lactis lactase also displays considerable activity at low temperature (5/sup 0/C). Enzyme stability is reduced drastically by demineralisation: addition of low concentrations of manganese ions (10/sup -3/ M) considerably enhances stability. Using a DDS Lab-Unit 35 fitted with GR61PP polysulphon membranes (cut-off: 20.000), pilot scale experiments were carried out (pH 6.5; 30/sup 0/C) in which whey permeate was hydrolyzed to a degree of hydrolysis of 82% minimum. Enzyme recuperation amounted to 96.5% per batch, all enzyme activity loss being due to thermal inactivation. Microbiological examination of the enzymatic membrane reactor showed that growth of mcicroorganisms can largely be suppressed by working at lower temperature (5/sup 0/C). Eventually, 50 ppm H/sub 2/O/sub 2/ or sterile filtration will adequately solve microbiological problems without affecting enzyme activity.

  17. Comparison of the role that entropy has played in processes of non-enzymatic and enzymatic catalysis

    International Nuclear Information System (INIS)

    Dixon Pineda, Manuel Tomas

    2012-01-01

    The function that entropy has played is compared in processes of non-enzymatic and enzymatic catalysis. The processes followed are showed: the kinetics of the acid hydrolysis of 3-pentyl acetate and cyclopentyl acetate catalyzed by hydrochloric acid and enzymatic hydrolysis of ethyl acetate and γ-butyrolactone catalyzed by pig liver esterase. The activation parameters of Eyring were determined for each process and interpreted the contribution of the entropy of activation for catalysis in this type of model reactions. (author) [es

  18. Avaliação da influência da temperatura e do tratamento enzimático no comportamento reológico do suco de abacaxi pérola (Ananas Comosus L. merr. Evaluation of influence from temperature and enzymatic treatment in the rheological behavior of pineapple (Ananas Comosus L. merr. juice

    Directory of Open Access Journals (Sweden)

    Adriano Cesar Calandrini Braga

    2013-03-01

    Full Text Available O objetivo deste trabalho foi determinar o comportamento reológico do suco de abacaxi- pérola natural e tratado com enzimas pectinolíticas. As condições de tratamento enzimático foram otimizadas através de um planejamento experimental do tipo fatorial completo 2k, com três repetições do ponto central. Na avaliação do comportamento reológico foram utilizadas duas amostras submetidas a peneiramento (N e D, analisadas em quatro diferentes temperaturas (10; 25; 50 e 65 °C. As análises reológicas foram realizadas utilizando um viscosímetro de cilindros concêntricos Brookfield e os dados experimentais foram ajustados ao modelo de Mizrahi-Berk. A relação entre temperatura e viscosidade aparente foi descrita por uma equação tipo Arrhenius. A otimização da atividade enzimática indicou, através da análise de variância e da metodologia de superfície de resposta, que as variáveis temperatura e tempo de tratamento exerceram efeito estatisticamente significativo (pThe objective this study was determinate the rheological behavior of pineapple juice natural and treated with pectinolytic enzymes. Tests for optimization of enzyme activity used in treatment were realized using a complete factorial planning 2k, with three repetitions of center point and two independent variables (temperature and time of treatment. In evaluation of rheological behavior of pineapple juice, were utilized two samples submitted to sieving (N and D, analyzed in four different temperatures (10, 25, 50 e 65 °C. The rheological analyses were realized using a Brookfield concentric cylinders viscometer and the relationship in the temperature and apparent viscosity was described by an equation type Arrhenius. The experimental data were adjusted by the Mizrahi-Berk model. The optimization tests indicated by ANOVA and response surface methodology that the variables temperature and time of treatment have statistically significant effect (p<0,05 on concentration of

  19. Size effects on acid bisulfite pretreatment efficiency: multiple product yields in spent liquor and enzymatic digestibility of pretreated solids

    Science.gov (United States)

    Yalan Liu; Jinwu Wang; Michael P. Wolcott

    2017-01-01

    Currently, feedstock size effects on chemical pretreatment performance were not clear due to the complexity of the pretreatment process and multiple evaluation standards such as the sugar recovery in spent liquor or enzymatic digestibility. In this study, we evaluated the size effects by various ways: the sugar recovery and coproduct yields in spent liquor, the...

  20. Enzymatic Saccharification and Ethanol Fermentation of Reed Pretreated with Liquid Hot Water

    Directory of Open Access Journals (Sweden)

    Jie Lu

    2012-01-01

    Full Text Available Reed is a widespread-growing, inexpensive, and readily available lignocellulosic material source in northeast China. The objective of this study is to evaluate the liquid hot water (LHW pretreatment efficiency of reed based on the enzymatic digestibility and ethanol fermentability of water-insoluble solids (WISs from reed after the LHW pretreatment. Several variables in the LHW pretreatment and enzymatic hydrolysis process were optimized. The conversion of glucan to glucose and glucose concentrations are considered as response variables in different conditions. The optimum conditions for the LHW pretreatment of reed area temperature of 180°C for 20min and a solid-to-liquid ratio of 1 : 10. These optimum conditions for the LHW pretreatment of reed resulted in a cellulose conversion rate of 82.59% in the subsequent enzymatic hydrolysis at 50°C for 72 h with a cellulase loading of 30 filter paper unit per gram of oven-dried WIS. Increasing the pretreatment temperature resulted in a higher enzymatic digestibility of the WIS from reed. Separate hydrolysis and fermentation of WIS showed that the conversion of glucan to ethanol reached 99.5% of the theoretical yield. The LHW pretreatment of reed is a suitable method to acquire a high recovery of fermentable sugars and high ethanol conversion yield.

  1. Increased release of fermentable sugars from elephant grass by enzymatic hydrolysis in the presence of surfactants

    International Nuclear Information System (INIS)

    Menegol, Daiane; Scholl, Angélica Luisi; Fontana, Roselei Claudete; Dillon, Aldo José Pinheiro; Camassola, Marli

    2014-01-01

    Highlights: • Milling is an attractive method to enhance the enzymatic hydrolysis of biomass. • Surfactants improve the efficiency of lignocellulose enzymatic hydrolysis. • Pretreatment with NaOH, smaller particle size and Tween 80® were more efficient. - Abstract: In the search for renewable energy sources, elephant grass is an alternative substrate for ethanol production, but this substrate must be hydrolyzed by cellulases and xylanases to liberate fermentable sugars. During enzymatic hydrolysis, cellulase activity is reduced by the irreversible adsorption of cellulase onto cellulose, decreasing the rate of hydrolysis. Adding surfactants during hydrolysis can improve the process. The effects of Tween® and Triton® surfactants on the enzymatic hydrolysis of elephant grass were evaluated in this context. The data indicate that pretreatment with sodium hydroxide, along with a smaller particle size (0.075–0.152 mm) and the use of Tween 80®, increased the efficiency of releasing reducing sugars from pretreated elephant grass biomass. Thus, it is possible to reduce grinding costs in second-generation ethanol production through the use of surfactants, as they allow efficient hydrolysis of larger biomass particles

  2. Enzymatic oxidative biodegradation of nanoparticles: Mechanisms, significance and applications

    Energy Technology Data Exchange (ETDEWEB)

    Vlasova, Irina I. [Department of Environmental and Occupational Health, Center for Free Radical and Antioxidant Health, Graduate School of Public Health, University of Pittsburgh, Pittsburgh, PA 15219 (United States); Research Institute for Physico-Chemical Medicine, Federal Medico-Biological Agency, Moscow 119453 (Russian Federation); Kapralov, Alexandr A. [Department of Environmental and Occupational Health, Center for Free Radical and Antioxidant Health, Graduate School of Public Health, University of Pittsburgh, Pittsburgh, PA 15219 (United States); Michael, Zachary P.; Burkert, Seth C. [Department of Chemistry, University of Pittsburgh, Pittsburgh, PA 15260 (United States); Shurin, Michael R. [Department of Pathology, University of Pittsburgh Medical Center, Pittsburgh, PA 15261 (United States); Department of Immunology, University of Pittsburgh Medical Center, Pittsburgh, PA 15261 (United States); Star, Alexander [Department of Chemistry, University of Pittsburgh, Pittsburgh, PA 15260 (United States); Shvedova, Anna A., E-mail: ats@cdc.gov [Pathology and Physiology Research Branch, Health Effects Laboratory Division (HELD), National Institute for Occupational Safety and Health (NIOSH) and Department of Physiology and Pharmacology, West Virginia University, Morgantown, WV 26505 (United States); Kagan, Valerian E., E-mail: kagan@pitt.edu [Department of Environmental and Occupational Health, Center for Free Radical and Antioxidant Health, Graduate School of Public Health, University of Pittsburgh, Pittsburgh, PA 15219 (United States); Department of Chemistry, University of Pittsburgh, Pittsburgh, PA 15260 (United States); Departments of Pharmacology and Chemical Biology and Radiation Oncology, University of Pittsburgh, Pittsburgh, PA 15260 (United States)

    2016-05-15

    Biopersistence of carbon nanotubes, graphene oxide (GO) and several other types of carbonaceous nanomaterials is an essential determinant of their health effects. Successful biodegradation is one of the major factors defining the life span and biological responses to nanoparticles. Here, we review the role and contribution of different oxidative enzymes of inflammatory cells – myeloperoxidase, eosinophil peroxidase, lactoperoxidase, hemoglobin, and xanthine oxidase – to the reactions of nanoparticle biodegradation. We further focus on interactions of nanomaterials with hemoproteins dependent on the specific features of their physico-chemical and structural characteristics. Mechanistically, we highlight the significance of immobilized peroxidase reactive intermediates vs diffusible small molecule oxidants (hypochlorous and hypobromous acids) for the overall oxidative biodegradation process in neutrophils and eosinophils. We also accentuate the importance of peroxynitrite-driven pathways realized in macrophages via the engagement of NADPH oxidase- and NO synthase-triggered oxidative mechanisms. We consider possible involvement of oxidative machinery of other professional phagocytes such as microglial cells, myeloid-derived suppressor cells, in the context of biodegradation relevant to targeted drug delivery. We evaluate the importance of genetic factors and their manipulations for the enzymatic biodegradation in vivo. Finally, we emphasize a novel type of biodegradation realized via the activation of the “dormant” peroxidase activity of hemoproteins by the nano-surface. This is exemplified by the binding of GO to cyt c causing the unfolding and ‘unmasking’ of the peroxidase activity of the latter. We conclude with the strategies leading to safe by design carbonaceous nanoparticles with optimized characteristics for mechanism-based targeted delivery and regulatable life-span of drugs in circulation. - Highlights: • Nanoparticles can be degraded by

  3. Shape and topology optimization of enzymatic microreactors

    DEFF Research Database (Denmark)

    Pereira Rosinha, Ines

    for effective and cost efficient reactors for pharmaceutical processes forces the industry to search for better technologies. In biochemical engineering, the used reactor design in a given process is usually limited to a range of well-established configurations and layouts. Usually the implemented reactors...... in a chemical process do not always yield in the best reaction conditions.This thesis develops an innovative application of topology and shape optimization methods to achemical engineering problem. The main goal is to design a reactor according to the limitations of the reaction system by modifying the reactor...... configuration. In this thesis structural optimization methods were exclusively applied to enzymatic microreactors. The case studies were chosen such that they can be experimentally tested afterwards. In this way, the design of the reactor is customized to the reaction system and itcontributes to the reduction...

  4. PRETREATMENT OF LIGNOCELLULOSIC BIOMASS FOR ENZYMATIC HYDROLYSIS

    Directory of Open Access Journals (Sweden)

    Doan Thai Hoa

    2017-11-01

    Full Text Available The cost of raw materials continues to be a limiting factor in the production of bio-ethanol from traditional raw materials, such as sugar and starch. At the same time, there are large amount of agricultural residues as well as industrial wastes that are of low or negative value (due to costs of current effluent disposal methods. Dilute sulfuric acid pretreatment of elephant grass and wood residues for the enzymatic hydrolysis of cellulose has been investigated in this study.    Elephant grass (agricultural residue and sawdust (Pulp and Paper Industry waste with a small particulate size were treated using different dilute sulfuric acid concentrations at a temperature  of 140-170°C within 0.5-3 hours. The appropriate pretreatment conditions give the highest yield of soluble saccharides and total reducing sugars.

  5. Structure of the enzymatically synthesized fructan inulin

    International Nuclear Information System (INIS)

    Heyer, A.G.; Schroeer, B.; Radosta, S.; Wolff, D.; Czapla, S.; Springer, J.

    1998-01-01

    Construction, purification and characterization of a fusion protein of maltose-binding protein of Escherichia coli and the fructosyltransferase of Streptococcus mutans is described. With the purified protein, in vitro synthesis of inulin was performed. The obtained polysaccharide was characterized by high-performance size-exclusion chromatography (HPSEC) and static light scattering (SLS) in dilute aqueous and dimethyl sulfoxide solution. For all samples very high molecular weights between 60x10 6 and 90x10 6 g/mol and a remarkable small polydispersity index of 1.1 have been determined. Small root-mean-square radii of gyration point to a compact conformation in dilute solution. No difference between native and enzymatically synthesized inulin was observed by X-ray powder diffraction and thermoanalysis of solid samples. (Copyright (c) 1998 Elsevier Science B.V., Amsterdam. All rights reserved.)

  6. Structure of the enzymatically synthesized fructan inulin

    Energy Technology Data Exchange (ETDEWEB)

    Heyer, A.G.; Schroeer, B. [Max-Planck-Institut fuer Molekulare Pflanzenphysiologie, Karl-Liebknecht-Str. 25, 14476 Golm (Germany); Radosta, S. [Fraunhofer-Institut fuer Angewandte Polymerforschung, Postfach 126, 14504 Teltow (Germany); Wolff, D.; Czapla, S.; Springer, J. [Technische Universitaet Berlin, FG Makromolekulare Chemie, Str. des 17. Juni 135, 10623 Berlin (Germany)

    1998-12-15

    Construction, purification and characterization of a fusion protein of maltose-binding protein of Escherichia coli and the fructosyltransferase of Streptococcus mutans is described. With the purified protein, in vitro synthesis of inulin was performed. The obtained polysaccharide was characterized by high-performance size-exclusion chromatography (HPSEC) and static light scattering (SLS) in dilute aqueous and dimethyl sulfoxide solution. For all samples very high molecular weights between 60x10{sup 6} and 90x10{sup 6} g/mol and a remarkable small polydispersity index of 1.1 have been determined. Small root-mean-square radii of gyration point to a compact conformation in dilute solution. No difference between native and enzymatically synthesized inulin was observed by X-ray powder diffraction and thermoanalysis of solid samples. (Copyright (c) 1998 Elsevier Science B.V., Amsterdam. All rights reserved.)

  7. Enzymatic hydrolysis of corn bran arabinoxylan

    DEFF Research Database (Denmark)

    Agger, Jane

    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...... in a complex and ridig cell wall structure. This thesis contains a thorough examination of the monosaccharide and structural composition of corn bran, which is used to assess and apply the relevant mono component enzyme preparations. In this way, the aim is to obtain the most effective minimal enzymatic......, especially with respect to xylose and glucose release, but vast amounts of the valuable monosaccharides are lost during this pretreatment and this is especially evident for arabinose. From a scientific point of view acid catalysed pretreatment renders the substrate in a state of disruption where assessment...

  8. Direct electron transfer based enzymatic fuel cells

    International Nuclear Information System (INIS)

    Falk, Magnus; Blum, Zoltan; Shleev, Sergey

    2012-01-01

    In this mini-review we briefly describe some historical developments made in the field of enzymatic fuel cells (FCs), discussing important design considerations taken when constructing mediator-, cofactor-, and membrane-less biological FCs (BFCs). Since the topic is rather extensive, only BFCs utilizing direct electron transfer (DET) reactions on both the anodic and cathodic sides are considered. Moreover, the performance of mostly glucose/oxygen biodevices is analyzed and compared. We also present some unpublished results on mediator-, cofactor-, and membrane-less glucose/oxygen BFCs recently designed in our group and tested in different human physiological fluids, such as blood, plasma, saliva, and tears. Finally, further perspectives for BFC applications are highlighted.

  9. Isothermal calorimetry on enzymatic biodiesel production

    DEFF Research Database (Denmark)

    Fjerbæk, Lene

    2008-01-01

    information about effects taking place when using lipases immobilized on an inert carrier for transesterification of a triglyceride and an alcohol as for biodiesel production. The biodiesel is produced by rapeseed oil and methanol as well as ethanol and a commercial biocatalyst Novozym 435 from Novozymes...... containing a Candida Antarctica B lipase immobilized on an acrylic resin. The reaction investigated is characterized by immiscible liquids (oil, methanol, glycerol and biodiesel) and enzymes imm. on an inert carrier during reaction, which allows several effects to take place that during normal reaction...... conditions can not be elucidated. These effects have been observed with isothermal calorimetry bringing forth new information about the reaction of enzymes catalyzing transesterification. Enzymatic biodiesel production has until now not been investigated with isothermal microcalorimetry, but the results...

  10. Cascade enzymatic reactions for efficient carbon sequestration.

    Science.gov (United States)

    Xia, Shunxiang; Zhao, Xueyan; Frigo-Vaz, Benjamin; Zheng, Wenyun; Kim, Jungbae; Wang, Ping

    2015-04-01

    Thermochemical processes developed for carbon capture and storage (CCS) offer high carbon capture capacities, but are generally hampered by low energy efficiency. Reversible cascade enzyme reactions are examined in this work for energy-efficient carbon sequestration. By integrating the reactions of two key enzymes of RTCA cycle, isocitrate dehydrogenase and aconitase, we demonstrate that intensified carbon capture can be realized through such cascade enzymatic reactions. Experiments show that enhanced thermodynamic driving force for carbon conversion can be attained via pH control under ambient conditions, and that the cascade reactions have the potential to capture 0.5 mol carbon at pH 6 for each mole of substrate applied. Overall it manifests that the carbon capture capacity of biocatalytic reactions, in addition to be energy efficient, can also be ultimately intensified to approach those realized with chemical absorbents such as MEA. Copyright © 2015 Elsevier Ltd. All rights reserved.

  11. Rapid enzymatic analysis of plasma for tyrosine.

    Science.gov (United States)

    Shimizu, H; Taniguchi, K; Sugiyama, M; Kanno, T

    1990-01-01

    In this rapid, simple, and convenient enzymatic method for measurement of tyrosine in plasma, tyrosine is converted to tyramine by action of tyrosine decarboxylase (EC 4.1.1.25) and the tyramine produced is oxidized to p-hydroxybenzyl aldehyde and hydrogen peroxide by action of tyramine oxidase (EC 1.4.3.9). The hydrogen peroxide is reacted with 4-aminoantipyrine and N-ethyl-N-(2-hydroxy-3-sulfopropyl)-m-toluidine in the presence of peroxidase (EC 1.11.1.7) to obtain quinoneimine dye, the absorbance of which is measured at 570 nm. Thus tyrosine is measured in the visible range. The CV was 4.6% or less, and the measurement was unaffected by other amino acids, except for phenylalanine. The values obtained (y) correlated well with those obtained with an amino acid analyzer (x): y = 0.902x + 3.92 mumol/L (Syx = 12.3; r = 0.985; n = 54).

  12. Radiation degradation and the subsequent enzymatic hydrolysis of waste paper

    International Nuclear Information System (INIS)

    Kamakura, M.; Kaetsu, I.

    1982-01-01

    Various studies have been carried out to find methods for the pretreatment of waste cellulosic materials to make them more susceptible to enzymatic hydrolysis. In the work reported here, the effects of preirradiating waste papers on subsequent enzymatic hydrolysis have been studied

  13. Enzymatic network for production of ether amines from alcohols

    NARCIS (Netherlands)

    Palacio, Cyntia M.; Crismaru, Gica Ciprian; Bartsch, Sebastian; Navickas, Vaidotas; Ditrich, Klaus; Breuer, Michael; Abu, Rohana; Woodley, John; Baldenius, Kai-Uwe; Wu, Bian; Janssen, Dick

    We constructed an enzymatic network composed of three different enzymes for the synthesis of valuable ether amines. The enzymatic reactions are interconnected to catalyze the oxidation and subsequent transamination of the substrate and to provide cofactor recycling. This allows production of the

  14. The Enzymatic Oxidation of Graphene Oxide

    Science.gov (United States)

    Kotchey, Gregg P.; Allen, Brett L.; Vedala, Harindra; Yanamala, Naveena; Kapralov, Alexander A.; Tyurina, Yulia Y.; Klein-Seetharaman, Judith; Kagan, Valerian E.; Star, Alexander

    2011-01-01

    Two-dimensional graphitic carbon is a new material with many emerging applications, and studying its chemical properties is an important goal. Here, we reported a new phenomenon – the enzymatic oxidation of a single layer of graphitic carbon by horseradish peroxidase (HRP). In the presence of low concentrations of hydrogen peroxide (~40 µM), HRP catalyzed the oxidation of graphene oxide, which resulted in the formation of holes on its basal plane. During the same period of analysis, HRP failed to oxidize chemically reduced graphene oxide (RGO). The enzymatic oxidation was characterized by Raman, UV-Vis, EPR and FT-IR spectroscopy, TEM, AFM, SDS-PAGE, and GC-MS. Computational docking studies indicated that HRP was preferentially bound to the basal plane rather than the edge for both graphene oxide and RGO. Due to the more dynamic nature of HRP on graphene oxide, the heme active site of HRP was in closer proximity to graphene oxide compared to RGO, thereby facilitating the oxidation of the basal plane of graphene oxide. We also studied the electronic properties of the reduced intermediate product, holey reduced graphene oxide (hRGO), using field-effect transistor (FET) measurements. While RGO exhibited a V-shaped transfer characteristic similar to a single layer of graphene that was attributed to its zero band gap, hRGO demonstrated a p-type semiconducting behavior with a positive shift in the Dirac points. This p-type behavior rendered hRGO, which can be conceptualized as interconnected graphene nanoribbons, as a potentially attractive material for FET sensors. PMID:21344859

  15. Characterizing Enzymatic Deposition for Microelectrode Neurotransmitter Detection

    Energy Technology Data Exchange (ETDEWEB)

    Hosein, W. K. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Yorita, A. M. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Tolosa, V. M. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)

    2016-08-12

    The enzyme immobilization process, one step in creating an enzymatic biosensor, was characterized and analyzed as a function of its physical properties. The neural glutamic biosensor is a flexible device, effectively minimizing trauma to the area of implantation. The Multielectrode Array (MEA) is composed primarily of a proprietary polymer which has been successfully implanted into human subjects in recent years. This polymer allows the device the pliability that other devices normally lack, though this poses some challenges to implantation. The electrodes are made of Platinum (Pt), and can range in number from eight to thirty two electrodes per device. These electrodes are electroplated with a semipermeable polymer layer to improve selectivity of the electrode to the neurotransmitter of interest, in this case glutamate. A signal is created from the interaction of glutamate in the brain with the glutamate oxidase (GluOx) which is immobilized on the surface of the electrode by using crosslinking chemistry in conjunction with glutaraldehyde and Bovine Serum Albumin (BSA). The glutamate is oxidized by glutamate oxidase, producing α-ketoglutarate and hydrogen peroxide (H2O2) as a by-product. The production of H2O2 is crucial for detection of the presence of the glutamate within the enzymatic coating, as it diffuses through the enzyme layer and oxidizes at the surface of the electrode. This oxidation is detectable by measurable change in the current using amperometry. Hence, the MEA allows for in vivo monitoring of neurotransmitter activity in real time. The sensitivity of the sensor to these neurotransmitters is dependent on the thickness of the layer, which is investigated in these experiments in order to optimize the efficacy of the device to detecting the substrate, once implanted.

  16. Quantitative determination of five metabolites of aspirin by UHPLC-MS/MS coupled with enzymatic reaction and its application to evaluate the effects of aspirin dosage on the metabolic profile.

    Science.gov (United States)

    Li, Jian-Ping; Guo, Jian-Ming; Shang, Er-Xin; Zhu, Zhen-Hua; Liu, Yang; Zhao, Bu-Chang; Zhao, Jing; Tang, Zhi-Shu; Duan, Jin-Ao

    2017-05-10

    Acetylsalicylic acid (Aspirin, ASA) is a famous drug for cardiovascular diseases in recent years. Effects of ASA dosage on the metabolic profile have not been fully understood. The purpose of our study is to establish a rapid and reliable method to quantify ASA metabolites in biological matrices, especially for glucuronide metabolites whose standards are not commercially available. Then we applied this method to evaluate the effects of ASA dosage on the metabolic and excretion profile of ASA metabolites in rat urine. Salicylic acid (SA), gentisic acid (GA) and salicyluric acid (SUA) were determined directly by UHPLC-MS/MS, while salicyl phenolic glucuronide (SAPG) and salicyluric acid phenolic glucuronide (SUAPG) were quantified indirectly by measuring the released SA and SUA from SAPG and SUAPG after β-glucuronidase digestion. SUA and SUAPG were the major metabolites of ASA in rat urine 24h after ASA administration, which accounted for 50% (SUA) and 26% (SUAPG). When ASA dosage was increased, the contributions dropped to 32% and 18%, respectively. The excretion of other three metabolites (GA, SA and SAPG) however showed remarkable increases by 16%, 6% and 4%, respectively. In addition, SUA and SUAPG were mainly excreted in the time period of 12-24h, while GA was excreted in the earlier time periods (0-4h and 4-8h). SA was mainly excreted in the time period of 0-4h and 12-24h. And the excretion of SAPG was equally distributed in the four time periods. We went further to show that the excretion of five metabolites in rat urine was delayed when ASA dosage was increased. In conclusion, we have developed a rapid and sensitive method to determine the five ASA metabolites (SA, GA, SUA, SAPG and SUAPG) in rat urine. We showed that ASA dosage could significantly influence the metabolic and excretion profile of ASA metabolites in rat urine. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Enzymatic hydrolysis of biomimetic bacterial cellulose-hemicellulose composites.

    Science.gov (United States)

    Penttilä, Paavo A; Imai, Tomoya; Hemming, Jarl; Willför, Stefan; Sugiyama, Junji

    2018-06-15

    The production of biofuels and other chemicals from lignocellulosic biomass is limited by the inefficiency of enzymatic hydrolysis. Here a biomimetic composite material consisting of bacterial cellulose and wood-based hemicelluloses was used to study the effects of hemicelluloses on the enzymatic hydrolysis with a commercial cellulase mixture. Bacterial cellulose synthesized in the presence of hemicelluloses, especially xylan, was found to be more susceptible to enzymatic hydrolysis than hemicellulose-free bacterial cellulose. The reason for the easier hydrolysis could be related to the nanoscale structure of the substrate, particularly the packing of cellulose microfibrils into ribbons or bundles. In addition, small-angle X-ray scattering was used to show that the average nanoscale morphology of bacterial cellulose remained unchanged during the enzymatic hydrolysis. The reported easier enzymatic hydrolysis of bacterial cellulose produced in the presence of wood-based xylan offers new insights to overcome biomass recalcitrance through genetic engineering. Copyright © 2018 Elsevier Ltd. All rights reserved.

  18. Enzymatic browning and after-cooking darkening of Jerusalem artichoke tubers (Helianthus tuberosus L.)

    DEFF Research Database (Denmark)

    Bach, Vibe; Bennedbæk-Jensen, Sidsel; Clausen, Morten Rahr

    2013-01-01

    Jerusalem artichoke tubers (Helianthus tuberosus L.) undergo enzymatic browning when peeled or cut, and turn grey after boiling, due to after-cooking darkening reactions between iron and phenolic acids. In an attempt to reveal the components responsible for these discolouration reactions, sensory...... evaluation and instrumental colour measurements were related to contents of total phenolics, phenolic acids, organic acids and iron in three varieties of raw and boiled Jerusalem artichoke tubers harvested in the autumn and the spring. No differences were found between varieties in sensory evaluated...... enzymatic browning, but Rema and Draga had higher scores than Mari in after-cooking darkening. Jerusalem artichoke tubers had higher contents of total phenolics, phenolic acids and citric acid in the autumn and low contents in the spring, while it was the opposite for malic acid. None of the chemical...

  19. Enzymatic browning and after-cooking darkening of Jerusalem artichoke tubers (Helianthus tuberosus L.).

    Science.gov (United States)

    Bach, Vibe; Jensen, Sidsel; Clausen, Morten R; Bertram, Hanne C; Edelenbos, Merete

    2013-11-15

    Jerusalem artichoke tubers (Helianthus tuberosus L.) undergo enzymatic browning when peeled or cut, and turn grey after boiling, due to after-cooking darkening reactions between iron and phenolic acids. In an attempt to reveal the components responsible for these discolouration reactions, sensory evaluation and instrumental colour measurements were related to contents of total phenolics, phenolic acids, organic acids and iron in three varieties of raw and boiled Jerusalem artichoke tubers harvested in the autumn and the spring. No differences were found between varieties in sensory evaluated enzymatic browning, but Rema and Draga had higher scores than Mari in after-cooking darkening. Jerusalem artichoke tubers had higher contents of total phenolics, phenolic acids and citric acid in the autumn and low contents in the spring, while it was the opposite for malic acid. None of the chemical parameters investigated could explain the discolouration of the Jerusalem artichoke tubers. Copyright © 2013 Elsevier Ltd. All rights reserved.

  20. Reduced need for grafting and low incidence of hypertrophic scarring in burns after enzymatic debridement

    OpenAIRE

    Corrales-Benítez, Carlos; Martínez-Méndez, José Ramón; González-Miranda, Álvaro; Serrano-Alonso, Maite; Casado-Pérez, César

    2016-01-01

    Background and Objective. Early burn scar removal is the standard of care for burn patients; excisional debridement, however, often leads to excision of dermal remnants, making autografting unavoidable. Enzymatic debridement with proteolytic enzymes enriched in Bromelain (Nexobrid®) leaves these dermal remnants to allow spontaneous healing of partial thickness burns. This study evaluates if Nexobrid® (MediWound Ltd., Israel) reduces the need for surgery and autografting in intermediate and de...

  1. Production of structured lipid with a low omega-6/omega-3 fatty acids ratio by enzymatic interesterification; Producción de lípidos estructurados con una baja relación de ácidos grasos omega-6/omega-3 mediante interesterificación enzimática.

    Energy Technology Data Exchange (ETDEWEB)

    Ilyasoglu, H.

    2017-07-01

    A structured lipid (SL) constituting omega fatty acids was synthesized by using linseed and grape seed oils as substrates via a lipase-catalyzed reaction. Lipozyme® TL IM was used as a biocatalyst. Good quadratic models predicting the incorporation of omega fatty acids were achieved via the Response surface methodology (RSM). The optimal conditions for targeted omega-6/omega-3 fatty acid ratio (2:1) were obtained at a substrate molar ratio 1.4, time 8.4 h, and enzyme amount 6.4%. The SL contained linoleic acid (43 g 100g-1), which was mainly located in the sn-2 position (40 g 100g-1). α-Linoleic acid, and α-linolenic acid at the sn-2 position were 22 g 100g-1, and 11 g 100g-1, respectively. The oxidative stability of the SL, and SL with antioxidants was also investigated. The produced SL may be proposed as a source of a balanced intake of omega fatty acids and an ingredient in functional food formulations. [Spanish] Se sintetizaron lípidos estructurados (SL), formados por ácidos grasos omega, utilizando aceites de linaza y semillas de uva como sustratos a través de una reacción catalizada por lipasa. Se utilizó Lipozyme® TL IM como biocatalizador. Los buenos modelos cuadráticos que predecían la incorporación de los ácidos grasos omega se lograron a través de la metodología de superficie de respuesta (RSM). Se obtuvieron las condiciones óptimas para una proporción de ácidos grasos omega-6/omega-3 (2:1) con una relación molar de sustrato 1:4, tiempo de 8,4 h, y cantidad de enzima 6,4%. El SL contenía ácido linoleico (43 g·100 g-1), que se localizaba principalmente en la posición sn-2 (40 g·100 g-1). El ácido α-linoleico y el ácido α-linolénico en la posición sn-2 fueron de 22 g·100 g-1y 11 g·100 g-1, respectivamente. También se investigó la estabilidad oxidativa del SL y SL con antioxidantes. El SL producido puede ser propuesto como una fuente para una ingesta equilibrada de ácidos grasos omega y un ingrediente en las formulaciones de alimentos funcionales.

  2. evaluation of microbial and enzymatic communities in soil

    African Journals Online (AJOL)

    ACSS

    2018-02-13

    Feb 13, 2018 ... The β-glucosidase, acid phosphatase and arylsulphatase activities were determined ... In view of these results, it is possible that the yield of soybean plants influences the number of .... Adapted from Santos et al. (2017) ...

  3. Application of extended Kalman filter to identification of enzymatic deactivation.

    Science.gov (United States)

    Caminal, G; Lafuente, J; López-Santín, J; Poch, M; Solà, C

    1987-02-01

    A recursive estimation scheme, the Extended Kalman Filter (EKF) technique, was applied to study enzymatic deactivation in the enzymatic hydrolysis of pretreated cellulose using a model previously developed by the authors. When no deactivation model was assumed, the results showed no variation with time for all the model parameters except for the maximum rate of cellobiose-to-glucose conversion (r'(m)).The r'(m) variation occurred in two zones with a grace period. A new model of enzymatic hydrolysis of pretreated cellulose deactivation was proposed and validated showing better behavior than the old deactivation model. This approach allows one to study enzyme deactivation without additional experiments and within operational conditions.

  4. Aqueous enzymatic extraction of Moringa oleifera oil.

    Science.gov (United States)

    Mat Yusoff, Masni; Gordon, Michael H; Ezeh, Onyinye; Niranjan, Keshavan

    2016-11-15

    This paper reports on the extraction of Moringa oleifera (MO) oil by using aqueous enzymatic extraction (AEE) method. The effect of different process parameters on the oil recovery was discovered by using statistical optimization, besides the effect of selected parameters on the formation of its oil-in-water cream emulsions. Within the pre-determined ranges, the use of pH 4.5, moisture/kernel ratio of 8:1 (w/w), and 300stroke/min shaking speed at 40°C for 1h incubation time resulted in highest oil recovery of approximately 70% (goil/g solvent-extracted oil). These optimized parameters also result in a very thin emulsion layer, indicating minute amount of emulsion formed. Zero oil recovery with thick emulsion were observed when the used aqueous phase was re-utilized for another AEE process. The findings suggest that the critical selection of AEE parameters is key to high oil recovery with minimum emulsion formation thereby lowering the load on the de-emulsification step. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. Electrochemical non-enzymatic glucose sensors

    International Nuclear Information System (INIS)

    Park, Sejin; Boo, Hankil; Chung, Taek Dong

    2006-01-01

    The electrochemical determination of glucose concentration without using enzyme is one of the dreams that many researchers have been trying to make come true. As new materials have been reported and more knowledge on detailed mechanism of glucose oxidation has been unveiled, the non-enzymatic glucose sensor keeps coming closer to practical applications. Recent reports strongly imply that this progress will be accelerated in 'nanoera'. This article reviews the history of unraveling the mechanism of direct electrochemical oxidation of glucose and making attempts to develop successful electrochemical glucose sensors. The electrochemical oxidation of glucose molecules involves complex processes of adsorption, electron transfer, and subsequent chemical rearrangement, which are combined with the surface reactions on the metal surfaces. The information about the direct oxidation of glucose on solid-state surfaces as well as new electrode materials will lead us to possible breakthroughs in designing the enzymeless glucose sensing devices that realize innovative and powerful detection. An example of those is to introduce nanoporous platinum as an electrode, on which glucose is oxidized electrochemically with remarkable sensitivity and selectivity. Better model of such glucose sensors is sought by summarizing and revisiting the previous reports on the electrochemistry of glucose itself and new electrode materials

  6. Enzymatic Systems for Cellulose Acetate Degradation

    Directory of Open Access Journals (Sweden)

    Oskar Haske-Cornelius

    2017-09-01

    Full Text Available Cellulose acetate (CA-based materials, like cigarette filters, contribute to landscape pollution challenging municipal authorities and manufacturers. This study investigates the potential of enzymes to degrade CA and to be potentially incorporated into the respective materials, enhancing biodegradation. Deacetylation studies based on Liquid Chromatography-Mass Spectrometry-Time of Flight (LC-MS-TOF, High Performance Liquid Chromatography (HPLC, and spectrophotometric analysis showed that the tested esterases were able to deacetylate the plasticizer triacetin (glycerol triacetate and glucose pentaacetate (cellulose acetate model compound. The most effective esterases for deacetylation belong to the enzyme family 2 (AXE55, AXE 53, GAE, they deacetylated CA with a degree of acetylation of up to 1.8. A combination of esterases and cellulases showed synergistic effects, the absolute glucose recovery for CA 1.8 was increased from 15% to 28% when an enzymatic deacetylation was performed. Lytic polysaccharide monooxygenase (LPMO, and cellobiohydrolase were able to cleave cellulose acetates with a degree of acetylation of up to 1.4, whereas chitinase showed no activity. In general, the degree of substitution, chain length, and acetyl group distribution were found to affect CA degradation. This study shows that, for a successful enzyme-based deacetylation system, a cocktail of enzymes, which will randomly cleave and generate shorter CA fragments, is the most suitable.

  7. Method for the enzymatic production of hydrogen

    Science.gov (United States)

    Woodward, J.; Mattingly, S.M.

    1999-08-24

    The present invention is an enzymatic method for producing hydrogen comprising the steps of: (a) forming a reaction mixture within a reaction vessel comprising a substrate capable of undergoing oxidation within a catabolic reaction, such as glucose, galactose, xylose, mannose, sucrose, lactose, cellulose, xylan and starch; the reaction mixture also comprising an amount of glucose dehydrogenase in an amount sufficient to catalyze the oxidation of the substrate, an amount of hydrogenase sufficient to catalyze an electron-requiring reaction wherein a stoichiometric yield of hydrogen is produced, an amount of pH buffer in an amount sufficient to provide an environment that allows the hydrogenase and the glucose dehydrogenase to retain sufficient activity for the production of hydrogen to occur and also comprising an amount of nicotinamide adenine dinucleotide phosphate sufficient to transfer electrons from the catabolic reaction to the electron-requiring reaction; (b) heating the reaction mixture at a temperature sufficient for glucose dehydrogenase and the hydrogenase to retain sufficient activity and sufficient for the production of hydrogen to occur, and heating for a period of time that continues until the hydrogen is no longer produced by the reaction mixture, wherein the catabolic reaction and the electron-requiring reactions have rates of reaction dependent upon the temperature; and (c) detecting the hydrogen produced from the reaction mixture. 8 figs.

  8. THEORY DEVELOPMENT OF ENZYMATIC AROMA RECOVERY

    Directory of Open Access Journals (Sweden)

    G. E. Dubova

    2014-01-01

    Full Text Available Summary. The fruit and vegetable pretreatment conditions and subsequent environment in which enzymatic reactions take place can be considered as potential factors in the formation of fresh flavors. The synthesis of aromatic components of fresh grass and green leaves occurs involving vegetable lipoxygenases. The molecules of a precursor-compound can withstand the processing modes, while enzymes and aromatic compounds break down frequently. Vegetable homogenates are potential sources of enzymes which produce natural aromatic substances. Formation of fresh favors is the most perceptible when it occurs as the result of the reaction between poliunsaturated fatty acids of cytoplasmic membranes and lipoxygenases and hydroperoxide lyase of plant material. Pre-treatment of samples positively influences binding energy in the complex of enzyme-substrate. The change of iodine number in treated homogenates, as compared to fresh ones, shows isomerization of flavor precursors. The minimal quantity of homogenates introduced (up to 20 g and the duration of aroma-restoring reaction (from 5 to 7 minutes were defined. Pre-cooling of homogenates activates enzymes, strengthens oxidability of the PUFA, and results in recovery of fresh aroma of plant material. Under conditions of enzyme inactivation, the synthesis of aromas is not possible. Conversely, production of aroma in food glazes and foams is possible in case of interphase activation between a substrate and enzymes.

  9. Effects of surface proteins and lipids on molecular structure, thermal properties, and enzymatic hydrolysis of rice starch

    Directory of Open Access Journals (Sweden)

    Pan HU

    Full Text Available Abstract Rice starches with different amylose contents were treated with sodium dodecyl sulfate (SDS to deplete surface proteins and lipids, and the changes in molecular structure, thermal properties, and enzymatic hydrolysis were evaluated. SDS treatment did not significantly change the molecular weight distribution, crystalline structure, short-range ordered degree, and gelatinization properties of starch, but significantly altered the pasting properties and increased the swelling power of starch. The removal of surface proteins and lipids increased the enzymatic hydrolysis and in vitro digestion of starch. The influences of removing surface proteins and lipids from starch on swelling power, pasting properties, and enzymatic hydrolysis were different among the various starches because of the differences in molecular structures of different starch styles. The aforementioned results indicated that removing the surface proteins and lipids from starch did not change the molecular structure but had significant effects on some functional properties.

  10. Hydrolysis of Miscanthus for bioethanol production using dilute acid presoaking combined with wet explosion pre-treatment and enzymatic treatment

    DEFF Research Database (Denmark)

    Sørensen, Annette; Teller, Philip Johan; Hilstrøm, Troels

    2008-01-01

    xylose prior to wet explosion. The acid presoaking extracted 63.2% xylose and 5.2% glucose. Direct enzymatic hydrolysis of the presoaked biomass was found to give only low sugar yields of 24-26% glucose. Wet explosion is a pre-treatment method that combines wet-oxidation and steam explosion. The effect...... of wet explosion on non-presoaked and presoaked Miscanthus was investigated using both atmospheric air and hydrogen peroxide as the oxidizing agent. All wet explosion pre-treatments showed to have a disrupting effect on the lignocellulosic biomass, making the sugars accessible for enzymatic hydrolysis......Miscanthus is a high yielding bioenergy crop. In this study we used acid presoaking, wet explosion, and enzymatic hydrolysis to evaluate the combination of the different pre-treatment methods for bioethanol production with Miscanthus. Acid presoaking is primarily carried out in order to remove...

  11. Comparison of Enzymatic and Ultrasonic Extraction of Albumin from Defatted Pumpkin (Cucurbita pepo)
Seed Powder.

    Science.gov (United States)

    Tu, Gia Loi; Bui, Thi Hoang Nga; Tran, Thi Thu Tra; Ton, Nu Minh Nguyet; Man Le, Van Viet

    2015-12-01

    In this study, ultrasound- and enzyme-assisted extractions of albumin (water-soluble protein group) from defatted pumpkin ( Cucurbita pepo ) seed powder were compared. Both advanced extraction techniques strongly increased the albumin yield in comparison with conventional extraction. The extraction rate was two times faster in the ultrasonic extraction than in the enzymatic extraction. However, the maximum albumin yield was 16% higher when using enzymatic extraction. Functional properties of the pumpkin seed albumin concentrates obtained using the enzymatic, ultrasonic and conventional methods were then evaluated. Use of hydrolase for degradation of cell wall of the plant material did not change the functional properties of the albumin concentrate in comparison with the conventional extraction. The ultrasonic extraction enhanced water-holding, oil-holding and emulsifying capacities of the pumpkin seed albumin concentrate, but slightly reduced the foaming capacity, and emulsion and foam stability.

  12. Onopordum nervosum as biomass source: some aspects of its production and transformation by enzymatic hydrolysis

    Energy Technology Data Exchange (ETDEWEB)

    Manzanares, P; Negro, M J; Saez, R; Martin, C [Centro de Investigaciones Energeticas, Medioambientales y Tecnologicas, Madrid (Spain). Inst. de Energias Renovables; Fernandez, J [ETSIA, Madrid (Spain). Dept. de Produccion Vegetal, Botanica y Proteccion Vegetal

    1993-01-01

    Onopordum nervosum, a lignocellulosic herbaceous species of the Iberian Peninsula, has been selected as a suitable biomass source to be used in transformation processes to obtain energy or industrial products. In this work, the effectiveness of different chemical pretreatments as a preliminary step to the enzymatic hydrolysis of this lignocellulosic biomass was evaluated. In order to determine biomass productivity, field assays were carried out in 1988 and 1989 using different planting densities and evaluating the effect to top fertilization. Biomass yields between 12 and 20 t ha[sup -1] were obtained, depending on the year and the planting density assayed. No significant differences were found in production rates when top fertilization was applied. Enzymatic hydrolysis of O.nervosum using a cellulolytic complex from Trichoderma longibrachiatum QM9414, gave low yields when untreated lignocellulosic biomass was used as substrate. Among different chemical pretreatments tested, ethanol and butanol solubilizations in the presence of a basic catalyst gave the best results. For the most effective pretreatment conditions, a delignification of about 30% and a complete recovery of glucose in the treated substrate were obtained both for butanol and ethanol. The highest enzymatic hydrolysis yields were found when ethanol was used as solvent, giving a saccharification efficiency of about 66% which, compared to the 23% for the native substrate, indicates the remarkable increment in the susceptibility of the cellulose to enzyme attack effected by this pretreatment. (author)

  13. Gamma radiation use to avoid enzymatic browning of cassava root (Manihot utilissima Pohl) in natura, peeled

    International Nuclear Information System (INIS)

    Costa Neto, Pedro Ramos da

    1997-01-01

    Cassava root was treated with gamma radiation at doses of 2, 4, 6, 8 and 10 kGy to avoid enzymatic browning. The irradiated samples were kept for 9 days at room temperature and evaluated for color and sensorial analysis. Two days after harvest, the control sample showed black spots and alterations of organoleptic characteristics. The irradiated sample with 2 to 6 kGy showed good appearance and acceptability. However, after 9 days of storage, the control and the irradiated (2 to 6 kGy) samples were not safe to eat, only the irradiated cassava with doses of 8 and 10 kGy did not show enzymatic browning and kept the good sensorial characteristics. (author)

  14. Enzymatic treatment of paper mill process waters; Entsyymit paperitehtaan kiertoveden kaesittelyssae - EKT 06

    Energy Technology Data Exchange (ETDEWEB)

    Mustranta, A; Buchert, J [VTT Biotechnology and Food Research, Espoo (Finland); Ekman, R; Spetz, P [Aabo Akademi, Turku (Finland). Lab. of Forest Products Chemistry; Luukko, K [Helsinki Univ. of Technology, Otaniemi (Finland). Paper Technology

    1999-12-31

    Dissolved and colloidal substances (DCS) are dispersed into the process waters during different stages of pulp and paper production. These are lipophilic extractives (pitch), hydrophilic extractives (lignan) and carbohydrates, mainly hemicelluloses. These dissolved and colloidal substances accumulate during water circulation and results in impaired paper machine runnability. DCS can also interfere with wet-end process chemicals. In this project the chemical composition of the process waters of spruce TMP pulping have been characterized. Simultaneously, potential enzymes for modification of DCS has been produced and purified. The enzymatic treatments have been started with lipase acting on triglycerides present in extractives. The effect of enzymatic treatment on the properties of process waters and technical properties of the pulp have been evaluated. (orig.)

  15. Enzymatic treatment of paper mill process waters; Entsyymit paperitehtaan kiertoveden kaesittelyssae - EKT 06

    Energy Technology Data Exchange (ETDEWEB)

    Mustranta, A.; Buchert, J. [VTT Biotechnology and Food Research, Espoo (Finland); Ekman, R.; Spetz, P. [Aabo Akademi, Turku (Finland). Lab. of Forest Products Chemistry; Luukko, K. [Helsinki Univ. of Technology, Otaniemi (Finland). Paper Technology

    1998-12-31

    Dissolved and colloidal substances (DCS) are dispersed into the process waters during different stages of pulp and paper production. These are lipophilic extractives (pitch), hydrophilic extractives (lignan) and carbohydrates, mainly hemicelluloses. These dissolved and colloidal substances accumulate during water circulation and results in impaired paper machine runnability. DCS can also interfere with wet-end process chemicals. In this project the chemical composition of the process waters of spruce TMP pulping have been characterized. Simultaneously, potential enzymes for modification of DCS has been produced and purified. The enzymatic treatments have been started with lipase acting on triglycerides present in extractives. The effect of enzymatic treatment on the properties of process waters and technical properties of the pulp have been evaluated. (orig.)

  16. ENZYMATIC HYDROLYSIS OF SWITCHGRASS AND COASTAL BERMUDA GRASS PRETREATED USING DIFFERENT CHEMICAL METHODS

    Directory of Open Access Journals (Sweden)

    Jiele Xu

    2011-06-01

    Full Text Available To investigate the effects of biomass feedstock and pretreatment method on the enzyme requirement during hydrolysis, swichgrass and coastal Bermuda grass pretreated using H2SO4, NaOH, and Ca(OH2 at the optimal conditions were subjected to enzymatic hydrolysis using two enzyme combinations: NS 50013 + NS 50010 and Cellic CTec + Cellic HTec. The enzyme loadings were optimized, and correlations between feedstock property, pretreatment strategy, and enzyme usage were evaluated. The results show that pretreatment methods resulting in greater lignin contents in the pretreated biomass were generally associated with higher enzyme requirements. More sugars could be recovered from alkaline-pretreated biomass during enzymatic hydrolysis due to the better carbohydrate preservation achieved at mild pretreatment temperatures. The cellulase enzyme, Cellic CTec, was more efficient in catalyzing the hydrolysis of coastal Bermuda grass, a feedstock more digestible than the pretreated swichgrass, following pretreatment with NaOH or Ca(OH2.

  17. Laboratory scale production of glucose syrup by the enzymatic ...

    African Journals Online (AJOL)

    Jen

    Laboratory scale production of glucose syrup by the enzymatic ... The industrial processing of starch to glucose, maltose and dextrin involves gelatinization, ... due to non-availability of appropriate technology and industry to harness these into.

  18. [Methods for enzymatic determination of triglycerides in liver homogenates].

    Science.gov (United States)

    Höhn, H; Gartzke, J; Burck, D

    1987-10-01

    An enzymatic method is described for the determination of triacylglycerols in liver homogenate. In contrast to usual methods, higher reliability and selectivity are achieved by omitting the extraction step.

  19. Recent insights in enzymatic synthesis of fructooligosaccharides from inulin.

    Science.gov (United States)

    Singh, Ram Sarup; Singh, Rupinder Pal; Kennedy, John F

    2016-04-01

    In the past few years, people are paying more attention to their dietary habits, and functional foods are playing a key role in maintaining the health of man. Prebiotics are considered as a main component of the functional foods which are usually composed of short chains of carbohydrates. Fructooligosaccharides (FOSs) are considered as one of the main group of prebiotics which have recognisable bifidogenic properties. FOSs are obtained either by extraction from various plant materials or by enzymatic synthesis from different substrates. Enzymatically, these can be obtained either from sucrose using fructosyltransferase or from inulin by endoinulinase. Inulin is a potent substrate for the enzymatic production of FOSs. This review article will provide an overview on the inulin as potent substrate, microbial sources of endoinulinases, enzymatic synthesis of FOSs from inulin, commercial status of FOSs, and their future perspectives. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. Enzymatic biosensors based on the use of metal oxide nanoparticles

    International Nuclear Information System (INIS)

    Shi, Xinhao; Gu, Wei; Li, Bingyu; Chen, Ningning; Zhao, Kai; Xian, Yuezhong

    2014-01-01

    Over the past decades, various techniques have been developed to obtain materials at a nanoscale level to design biosensors with high sensitivity, selectivity and efficiency. Metal oxide nanoparticles (MONPs) are of particular interests and have received much attention because of their unique physical, chemical and catalytic properties. This review summarizes the progress made in enzymatic biosensors based on the use of MONPs. Synthetic methods, strategies for immobilization, and the functions of MONPs in enzymatic biosensing systems are reviewed and discussed. The article is subdivided into sections on enzymatic biosensors based on (a) zinc oxide nanoparticles, (b) titanium oxide nanoparticles, (c) iron oxide nanoparticles, and (d) other metal oxide nanoparticles. While substantial advances have been made in MONPs-based enzymatic biosensors, their applications to real samples still lie ahead because issues such as reproducibility and sensor stability have to be solved. (author)

  1. Modelling of the enzymatic kinetically controlled synthesis of cephalexin

    NARCIS (Netherlands)

    Schroën, C.G.P.H.; Fretz, C.B.; Bruin, de V.H.; Berendsen, W.; Moody, H.M.; Roos, E.C.; Roon, van J.L.; Kroon, P.J.; Strubel, M.; Janssen, A.E.M.; Tramper, J.

    2002-01-01

    In this study the influence of diffusion limitation on enzymatic kinetically controlled cephalexin synthesis from phenylglycine amide and 7-aminodeacetoxycephalosporinic acid (7-ADCA) was investigated systematically. It was found that if diffusion limitation occurred, both the synthesis/hydrolysis

  2. Factors of enzymatic biodiesel production from sludge palm oil (SPO ...

    African Journals Online (AJOL)

    ika

    2013-07-31

    Jul 31, 2013 ... Biodiesel is a non-toxic, renewable and environmental friendly fuel. This study ... of biodiesel from sludge palm oil (SPO), a low-cost waste oil via enzymatic catalysis. ... Increasing energy crisis and environmental concerns by.

  3. Enzymatic labelling of. gamma. -globulin and insulin with iodine-125

    Energy Technology Data Exchange (ETDEWEB)

    Lucka, B; Russin, K [Institute of Nuclear Physics, Krakow (Poland)

    1979-01-01

    The parameters of enzymatic labelling of proteins with iodine 125 were examined. The manner and sequence of reagent addition, the effects of reagent concentration, reaction time and total Na/sup 125/I activity on the labelling yield were determined.

  4. Kinetics of enzymatic hydrolysis of methyl ricinoleate

    Directory of Open Access Journals (Sweden)

    Neeharika, T. S.V.R.

    2015-12-01

    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

  5. Nanocrystal Bioassembly: Asymmetry, Proximity, and Enzymatic Manipulation

    Energy Technology Data Exchange (ETDEWEB)

    Claridge, Shelley A. [Univ. of California, Berkeley, CA (United States)

    2008-05-01

    Research at the interface between biomolecules and inorganic nanocrystals has resulted in a great number of new discoveries. In part this arises from the synergistic duality of the system: biomolecules may act as self-assembly agents for organizing inorganic nanocrystals into functional materials; alternatively, nanocrystals may act as microscopic or spectroscopic labels for elucidating the behavior of complex biomolecular systems. However, success in either of these functions relies heavily uponthe ability to control the conjugation and assembly processes.In the work presented here, we first design a branched DNA scaffold which allows hybridization of DNA-nanocrystal monoconjugates to form discrete assemblies. Importantly, the asymmetry of the branched scaffold allows the formation of asymmetric2assemblies of nanocrystals. In the context of a self-assembled device, this can be considered a step toward the ability to engineer functionally distinct inputs and outputs.Next we develop an anion-exchange high performance liquid chromatography purification method which allows large gold nanocrystals attached to single strands of very short DNA to be purified. When two such complementary conjugates are hybridized, the large nanocrystals are brought into close proximity, allowing their plasmon resonances to couple. Such plasmon-coupled constructs are of interest both as optical interconnects for nanoscale devices and as `plasmon ruler? biomolecular probes.We then present an enzymatic ligation strategy for creating multi-nanoparticle building blocks for self-assembly. In constructing a nanoscale device, such a strategy would allow pre-assembly and purification of components; these constructs can also act as multi-label probes of single-stranded DNA conformational dynamics. Finally we demonstrate a simple proof-of-concept of a nanoparticle analog of the polymerase chain reaction.

  6. Enzymatic assays for the diagnosis of bradykinin-dependent angioedema.

    Directory of Open Access Journals (Sweden)

    Federica Defendi

    Full Text Available BACKGROUND: The kinins (primarily bradykinin, BK represent the mediators responsible for local increase of vascular permeability in hereditary angioedema (HAE, HAE I-II associated with alterations of the SERPING1 gene and HAE with normal C1-Inhibitor function (HAE-nC1INH. Besides C1-Inhibitor function and concentration, no biological assay of kinin metabolism is actually available to help physicians for the diagnosis of angioedema (AE. We describe enzymatic tests on the plasma for diagnosis of BK-dependent AE. METHODS: The plasma amidase assays are performed using the Pro-Phe-Arg-p-nitroanilide peptide substrate to evaluate the spontaneous amidase activity and the proenzyme activation. We analyzed data of 872 patients presenting with BK-dependent AE or BK-unrelated diseases, compared to 303 controls. Anti-high MW kininogen (HK immunoblot was achieved to confirm HK cleavage in exemplary samples. Reproducibility, repeatability, limit of blank, limit of detection, precision, linearity and receiver operating characteristics (ROC were used to calculate the diagnostic performance of the assays. RESULTS: Spontaneous amidase activity was significantly increased in all BK-dependent AE, associated with the acute phase of disease in HAE-nC1INH, but preserved in BK-unrelated disorders. The increase of the amidase activity was associated to HK proteolysis, indicating its relevance to identify kininogenase activity. The oestrogens, known for precipitating AE episodes, were found as triggers of enzymatic activity. Calculations from ROC curves gave the optimum diagnostic cut-off for women (9.3 nmol⋅min(-1⋅mL(-1, area under curve [AUC] 92.1%, sensitivity 80.0%, and specificity 90.1% and for men (6.6 nmol·min(-1⋅mL(-1, AUC 91.0%, sensitivity 87.0% and specificity 81.2%. CONCLUSION: The amidase assay represents a diagnostic tool to help physicians in the decision to distinguish between BK-related and -unrelated AE.

  7. Enzymatic Synthesis of Lignin-Based Concrete Dispersing Agents.

    Science.gov (United States)

    Jankowska, Dagmara; Heck, Tobias; Schubert, Mark; Yerlikaya, Alpaslan; Weymuth, Christophe; Rentsch, Daniel; Schober, Irene; Richter, Michael

    2018-03-15

    Lignin is the most abundant aromatic biopolymer, functioning as an integral component of woody materials. In its unmodified form it shows limited water solubility and is relatively unreactive, so biotechnological lignin valorisation for high-performance applications is greatly underexploited. Lignin can be obtained from the pulp and paper industry as a by-product. To expand its application, a new synthesis route to new dispersing agents for use as concrete additives was developed. The route is based on lignin functionalisation by enzymatic transformation. Screening of lignin-modifying systems resulted in functionalised lignin polymers with improved solubility in aqueous systems. Through grafting of sulfanilic acid or p-aminobenzoic acid by fungal laccases, lignin became soluble in water at pH≤4 or pH≤7, respectively. Products were analysed and evaluated in miniaturised application tests in cement paste and mortar. Their dispersing properties match the performance criteria of commercially available lignosulfonates. The study provides examples of new perspectives for the use of lignin. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Novel flavonolignan hybrid antioxidants: From enzymatic preparation to molecular rationalization.

    Science.gov (United States)

    Vavříková, Eva; Křen, Vladimír; Jezova-Kalachova, Lubica; Biler, Michal; Chantemargue, Benjamin; Pyszková, Michaela; Riva, Sergio; Kuzma, Marek; Valentová, Kateřina; Ulrichová, Jitka; Vrba, Jiří; Trouillas, Patrick; Vacek, Jan

    2017-02-15

    A series of antioxidants was designed and synthesized based on conjugation of the hepatoprotective flavonolignan silybin with l-ascorbic acid, trolox alcohol or tyrosol via a C 12 aliphatic linker. These hybrid molecules were prepared from 12-vinyl dodecanedioate-23-O-silybin using the enzymatic regioselective acylation procedure with Novozym 435 (lipase B) or with lipase PS. Voltammetric analyses showed that the silybin-ascorbic acid conjugate exhibited excellent electron donating ability, in comparison to the other conjugates. Free radical scavenging, antioxidant activities and cytoprotective action were evaluated. The silybin-ascorbic acid hybrid exhibited the best activities (IC 50  = 30.2 μM) in terms of lipid peroxidation inhibition. The promising protective action of the conjugate against lipid peroxidation can be attributed to modulated electron transfer abilities of both the silybin and ascorbate moieties, but also to the hydrophobic C 12 linker facilitating membrane insertion. This was supported experimentally and theoretically by density functional theory (DFT) and molecular dynamics (MD) calculations. The results presented here can be used in the further development of novel multipotent antioxidants and cytoprotective agents, in particular for substances acting at an aqueous/lipid interface. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  9. Inhibition of tyrosinase-mediated enzymatic browning by sulfite and natural alternatives

    NARCIS (Netherlands)

    Kuijpers, T.F.M.; Vincken, J.P.

    2013-01-01

    Although sulfite is widely used to counteract enzymatic browning, its mechanism has remained largely unknown. We describe a double inhibitory mechanism of sulfite on enzymatic browning, affecting both the enzymatic oxidation of phenols into o‑quinones, as well as the non‑enzymatic

  10. Bioelectrocatalytic NAD+/NADH inter-conversion: transformation of an enzymatic fuel cell into an enzymatic redox flow battery.

    Science.gov (United States)

    Quah, Timothy; Milton, Ross D; Abdellaoui, Sofiene; Minteer, Shelley D

    2017-07-25

    Diaphorase and a benzylpropylviologen redox polymer were combined to create a bioelectrode that can both oxidize NADH and reduce NAD + . We demonstrate how bioelectrocatalytic NAD + /NADH inter-conversion can transform a glucose/O 2 enzymatic fuel cell (EFC) with an open circuit potential (OCP) of 1.1 V into an enzymatic redox flow battery (ERFB), which can be rapidly recharged by operation as an EFC.

  11. Effect of the steam explosion pretreatment on enzymatic hydrolysis of eucalyptus wood and sweet sorghum baggages

    International Nuclear Information System (INIS)

    Negro, M. J.; Martinez, J. M.; Manero, J.; Saez, F.; Martin, C.

    1991-01-01

    The effect of steam explosion treatment on the enzymatic hydrolysis yield of two different lignocellulosic substrates is studied. Raw materials have been pretreated in a pilot plant designed to work in batch and equipped with a reactor vessel of 2 1 working volume where biomass was heated at the desired temperature and then exploded and recovered in a cyclone. Temperatures from 190 to 230 degree celsius and reaction times from 2 to 8 min. have been assayed. The efficiency of the steam explosion treatment has been evaluated on the composition of the lignocellulosic materials as well as on their enzymatic hydrolysis yield using a cellulolytic complex from T. reesel. Results show a high solubilization rate of hemicelluloses and variable losses of cellulose and lignin depending on the conditions tested. Enzymatic hydrolysis yields of both substrates experimented remarkable increments, corresponding the highest values obtained to 210 degree celsius; 2 min. and 21O degree celsius; 4 min. for sorghum bagasse and eucalyptus wood respectively. (Author) 13 refs

  12. Effect of the steam explosion pretreatment on enzymatic hydrolysis of eucalyptus wood and sweet sorghum bagasse

    International Nuclear Information System (INIS)

    Negro, M.J.; Martinez, J.M.; Manero, J.; Saez, F.; Martin, C.

    1990-01-01

    The effect of steam explosion treatment on the enzymatic hydrolysis yield of two different lignocellulosic substrates is studied. Raw materials have been pretreated in a pilot plant designed to work in batch and equiped with a reactor vessel of 2 1 working volume where biomass was heated at the desired temperature and then exploded and recovered in a cyclone. Temperatures from 190 to 230 o C and reaction times from 2 to 8 min. have been assayed. The efficiency of the steam explosion treatment has been evaluated on the composition of the lignocellulosic materials as well as on their enzymatic hydrolysis yield using a cellulolytic complex from T. reesei. Results show a high solubilization rate of hemicelluloses ands variable losses of cellulose and lignin depending on the conditions tested. Enzymatic hydrolysis yields of both substrates experimented remarkable increments, correspondig the highest values obtained to 210 o C; 2 min. and 210 o C; 4 min. for sorghum bagasse and eucaliptus wood respectivelly. (Author). 13 refs

  13. Nickel-functionalized reduced graphene oxide with polyaniline for non-enzymatic glucose sensing

    International Nuclear Information System (INIS)

    Zhang, Bing; He, Yu; Liu, Bingqian; Tang, Dianping

    2015-01-01

    We have developed a new class of organic–inorganic hybrid nanostructures based on the use of reduced graphene oxide (rGO), polyaniline, and a nickel metal nanostructure. It was applied to efficient non-enzymatic sensing of glucose based on its electrocatalytic oxidation. Scanning electron microscopy and energy-dispersive X-Ray were employed to characterize the material. It is shown that the doped polyaniline plays an important role in the formation of the hybrid nanostructures. Improved analytical performance is found when the hybrid nanostructures were placed on a glassy carbon electrode and used for non-enzymatic sensing of glucose at a typical working potential of +450 mV and a pH value of 13. Features include a fast response (∼2 s), high sensitivity (6,050 μA mM −1 cm −2 ), a linear range from 0.1 μM to 1.0 mM, and a low detection limit (0.08 μM). The response to glucose follows a Michaelis-Menten kinetic behavior, and the K M value was determined to be 0.241 μM. Reproducibility and specificity are acceptable. Fructose and maltose do not interfere significantly. Importantly, the methodology was validated and evaluated for the analysis of 15 spiked human serum specimens, receiving in a good accordance with the results obtained by the non-enzymatic glucose sensing and the commercialized personal glucose meter. (author)

  14. Genetic diversity in soybean genotypes using phenotypic characters and enzymatic markers.

    Science.gov (United States)

    Zambiazzi, E V; Bruzi, A T; Sales, A P; Borges, I M M; Guilherme, S R; Zuffo, A M; Lima, J G; Ribeiro, F O; Mendes, A E S; Godinho, S H M; Carvalho, M L M

    2017-09-21

    The objective of this study was to evaluate the genetic diversity of soybean cultivars by adopting phenotypic traits and enzymatic markers, the relative contribution of agronomic traits to diversity, as well as diversity between the level of technology used in soybean cultivars and genetic breeding programs in which cultivars were inserted. The experiments were conducted on the field at the Center for Scientific and Technological Development in crop-livestock production and the Electrophoresis Laboratory of Lavras Federal University. The agronomic traits adopted were grain yield, plant height, first legume insertion, plant lodging, the mass of one thousand seeds, and days for complete maturation, in which the Euclidean distance, grouped by Tocher and UPGMA criteria, was obtained. After electrophorese gels for enzymatic systems, dehydrogenase alcohol, esterase, superoxide dismutase, and peroxidase were performed. The genetic similarity estimative was also obtained between genotypes by the Jaccard coefficient with subsequent grouping by the UPGMA method. The formation of two groups was shown using phenotypic characters in the genetic diversity study and individually discriminating the cultivar 97R73 RR. The character with the greatest contribution to the genetic divergence was grain yield with contribution higher than 90.0%. To obtain six different groups, individually discriminating the cultivars CG 8166 RR, FPS Jupiter RR, and BRS MG 780 RR, enzymatic markers were used. Cultivars carrying the RR technology presented more divergence than conventional cultivars and IPRO cultivars.

  15. Structural Changes of Lignin after Liquid Hot Water Pretreatment and Its Effect on the Enzymatic Hydrolysis

    Directory of Open Access Journals (Sweden)

    Wen Wang

    2016-01-01

    Full Text Available During liquid hot water (LHW pretreatment, lignin is mostly retained in the pretreated biomass, and the changes in the chemical and structural characteristics of lignin should probably refer to re-/depolymerization, solubilization, or glass transition. The residual lignin could influence the effective enzymatic hydrolysis of cellulose. The pure lignin was used to evaluate the effect of LHW process on its structural and chemical features. The surface morphology of LHW-treated lignin observed with the scanning electron microscopy (SEM was more porous and irregular than that of untreated lignin. Compared to the untreated lignin, the surface area, total pore volume, and average pore size of LHW-treated lignin tested with the Brunner-Emmet-Teller (BET measurement were increased. FTIR analysis showed that the chemical structure of lignin was broken down in the LHW process. Additionally, the impact of untreated and treated lignin on the enzymatic hydrolysis of cellulose was also explored. The LHW-treated lignin had little impact on the cellulase adsorption and enzyme activities and somehow could improve the enzymatic hydrolysis of cellulose.

  16. Combined Mechanical Destruction and Alkaline Pretreatment of Wheat Straw for Enhanced Enzymatic Saccharification

    Directory of Open Access Journals (Sweden)

    Qianqian Wang

    2014-09-01

    Full Text Available Wheat straw was pretreated by combined mechanical destruction and alkaline pretreatments to enhance enzymatic saccharification. Four strategies were employed to evaluate the potential of wheat straw as a feedstock for fermentable sugar production. The effects of the pretreatments on the substrate morphology, size distribution, chemical composition, and cellulose crystallinity, along with the subsequent enzymatic digestibility, were investigated. Optical microscope images showed that mechanical pretreatment alone resulted in poor fiber defibrillation, wherein samples mostly consisted of rigid fiber bundles, while integrated mechanical destruction and alkaline pretreatment led to relatively good fiber defibrillation. Low temperature NaOH/urea pretreatment can fibrillate the rigid fiber bundles into a relatively loose network and alter the structure of the treated substrate to make cellulose more accessible. The glucan conversion rates were 77% and 95% for integrated mechanical destruction and alkaline pretreatments and mechanical destruction followed by low temperature NaOH/urea and ammonium/urea pretreatments, respectively, after 72 h of enzymatic hydrolysis with enzyme loadings of 10 FPU cellulase per g of oven-dry substrate.

  17. Blood parameters and enzymatic and oxidative activity in the liver of chickens fed with calcium anacardate

    Directory of Open Access Journals (Sweden)

    Carlos Eduardo Braga Cruz

    Full Text Available ABSTRACT The aim of this research was to evaluate the inclusion of calcium anacardate (CAC as a source of anacardic acid in the diet of broiler chickens on blood parameters, and enzymatic and oxidative activity in the liver. A total of 840 male chicks, one day old, were kept in a completely randomised experimental design, with six treatments and seven replications of 20 birds, totalling 140 birds per treatment. The treatments consisted of feed without the addition of growth promoter (GP, feed with GP, and feed with no GP and the addition of CAC at levels of 0.25, 0.50, 0.75 and 1%. The biochemical blood variables to be analysed were uric acid, total cholesterol, HDL, LDL, creatinine, AST, ALT, triglycerides, total erythrocytes, haemoglobin, haematocrit, mean corpuscular volume, corpuscular haemoglobin concentration, total plasma protein, total leukocytes, heterophils, lymphocytes, platelets and heterophil/lymphocyte ratio. The concentrations of superoxide dismutase, glutathione peroxidase and malondialdehyde were analysed for the enzymatic and oxidative parameters in the liver. There were no significant differences between treatments in the blood parameters or the enzymatic and oxidative activity in the liver of the chickens, demonstrating that the use of calcium anacardate as a source of anacardic acid is non-toxic, and does not affect these parameters.

  18. Effect of wheat and Miscanthus straw biochars on soil enzymatic activity, ecotoxicity, and plant yield

    Science.gov (United States)

    Mierzwa-Hersztek, Monika; Gondek, Krzysztof; Klimkowicz-Pawlas, Agnieszka; Baran, Agnieszka

    2017-07-01

    The variety of technological conditions and raw materials from which biochar is produced is the reason why its soil application may have different effects on soil properties and plant growth. The aim of this study was to evaluate the effect of the addition of wheat straw and Miscanthus giganteus straw (5 t DM ha-1) and biochar obtained from this materials in doses of 2.25 and 5 t DM ha-1 on soil enzymatic activity, soil ecotoxicity, and plant yield (perennial grass mixture with red clover). The research was carried out under field conditions on soil with the granulometric composition of loamy sand. No significant effect of biochar amendment on soil enzymatic activity was observed. The biochar-amended soil was toxic to Vibrio fischeri and exhibited low toxicity to Heterocypris incongruens. Application of wheat straw biochar and M. giganteus straw biochar in a dose of 5 t DM ha-1 contributed to an increase in plant biomass production by 2 and 14%, respectively, compared to the soil with mineral fertilisation. Biochars had a more adverse effect on soil enzymatic activity and soil ecotoxicity to H. incongruens and V. fischeri than non-converted wheat straw and M. giganteus straw, but significantly increased the grass crop yield.

  19. The Effect of Irradiation Treatment on the Non-Enzymatic Browning Reaction in Legume Seeds

    International Nuclear Information System (INIS)

    El-Niely, H.F.G.

    2013-01-01

    The present study was conducted to evaluate the effects of gamma irradiation treatment, at room temperature, on the non-enzymatic browning reaction (Millerd reaction products, MRPs) generated in soybeans, broad beans and dried peas seeds at dose levels of 10, 30 and 60 kGy and their effects on the chemical constituents, soluble protein, available lysine and in vitro protein digestibility. The formation of MRPs in the studied legumes was assayed by monitoring the formation of brown pigments (browning intensity) by spectrophotometric method. The results revealed that the chemical composition of irradiated legumes showed non-significant differences relative to the raw one. A dose dependent decrease in soluble proteins and available lysine in the three legumes were observed. The non-enzymatic browning reaction was significantly increased with increasing the radiation dose, which was proved by changes in browning index tests. At the same time, the in vitro protein digestibility was increased after irradiation up to 60 kGy. Irradiation of dried peas with 60 kGy produced higher browning index than the other legumes. A positive correlation was observed between the radiation dose and the browning index for soybeans (R2= 0.96), broad beans (R2 = 0.81) and dried peas (R2 = 0.97) which means that 96%, 81% and 97 of the variation in the incidence of non-enzymatic browning reaction in soybean, broad bean and dried peas, respectively, are due to the effect of irradiation treatments. The present study suggests that the formation of non-enzymatic browning reaction did not impair the nutritional quality of legumes, therefore, the process of irradiation was helpful in increasing the in vitro protein digestibility of studied legumes. These results clearly indicated that gamma irradiation processing at the studied doses can add valuable effects to the studied legumes

  20. ENZYMATIC AND NON-ENZYMATIC ANTIOXIDANT DEFENSE WITH ALZHEIMER DISEASE1

    Directory of Open Access Journals (Sweden)

    A. Vaisi-Raygani

    2007-07-01

    Full Text Available The etiopathogenesis of Alzheimer's disease (AD is still unclear.  However, long-term oxidative stress is believed to be one of the major contributing factors in progression of neuronal degeneration and decline of cognitive function in AD. In order to assess the presence of oxidative stress in AD, we examined the enzymatic activities of the erythrocyte Cu-Zn superoxide dismutase (Cu-Zn SOD, glutathione peroxidase (GSH-Px, catalase (CAT, and plasma level of total antioxidant status (TAS in AD and control groups (age and sex-matched. The results showed that the Cu-Zn SOD activity was significantly higher and the level of GSH-Px and TAS activities were significantly lower in AD subjects than that in the control group (2111 ± 324 U/grHb, 43.7 ± 11.6 U/grHb, and 1.17 ± 0.23 mmol/l compared with 1371 ± 211 U/grHb; t= -2.17, P = 0.036, 56.3 ± 9.5 U/grHb; t=3.8, P = 0.014, and 1.54±0.2 mmol/l; t=11.18, P < 0.001, respectively.  While, the erythrocyte CAT activity was lower in AD subjects compared to the control group, the difference was not statistically significant (t = 1.3, P = 0.15. These findings support the idea that the oxidative stress plays an important role in the pathogenesis underlying AD neurodegeneration. In addition, the enzymatic activity of the erythrocyte Cu-Zn SOD and GSH-Px and the plasma level of TAS can be used as a measure of the oxidative stress and a marker for pathological changes in the brain of patients with AD. 

  1. ASSOCIATION BETWEEN ENZYMATIC AND NON-ENZYMATIC ANTIOXIDANT DEFENSE WITH ALZHEIMER DISEASE

    Directory of Open Access Journals (Sweden)

    A. Vaisi-Raygani

    2008-04-01

    Full Text Available The etiopathogenesis of dementia in Alzheimer's disease (AD is still unclear. However, long-term oxidative stress is believed to be one of the major contributing factors in progression of neuronal degeneration and decline of cognitive function in AD. In order to assess the presence of oxidative stress in AD, we examined the enzymatic activities of the erythrocyte Cu-Zn superoxide dismutase (Cu-Zn SOD, glutathione peroxidase (GSH-Px, catalase (CAT, and plasma level of total antioxidant status (TAS in AD and control groups (age and sex-matched. The results showed that the Cu-Zn SOD activity was significantly higher and the level of GSH-Px and TAS activities were significantly lower in AD subjects than that in the control group (2111±324 U/grHb, 43.7±11.6 U/grHb, and 1.17 ±0.23 mmol/L compared with 1371±211 U/gHb; t= -2.17, p=0.036, 56.3±9.5 U/gHb; t=3.8, p=0.014, and 1.54±0.2 mmol/L; t=11.18, P<0.001, respectively. While, the erythrocyte CAT activity was lower in AD subjects compared to the control group, the difference was not statistically significant (t=1.3, P=0.15. These findings support the idea that the oxidative stress plays an important role in the pathogenesis underlying AD neurodegeneration. In addition, the enzymatic activity of the erythrocyte Cu-Zn SOD and GSH-Px and the plasma level of TAS can be used as a measure of the oxidative stress and a marker for pathological changes in the brain of patients with AD.

  2. Effect of topical application of fluoride gel NaF 2% on enzymatic and non-enzymatic antioxidant parameters of saliva.

    Science.gov (United States)

    Leite, Mariana Ferreira; Ferreira, Nayara Ferraz D'Assumpção; Shitsuka, Caleb David Willy Moreira; Lima, Amanda Martins; Masuyama, Mônica Miyuki; Sant'Anna, Giselle Rodrigues; Yamaguti, Paula Mochidome; Polotow, Tatiana G; de Barros, Marcelo Paes

    2012-06-01

    The aim of the study was to evaluate the effect of topical fluoride gel NaF 2% application on antioxidant parameters of whole saliva from children. The saliva mechanically stimulated with parafilm was collected from 25 children (6-12 years) attending the Clinic of Paediatric Dentistry of Universidade Cruzeiro do Sul, São Paulo, Brazil, before (control group) and immediately after application of neutral fluoride gel NaF 2% (fluoride-gel group), according to the Standards for Research Using Human Subjects, Resolution 196/96 of the USA National Health Council of 10/10/1996. Afterwards, pre-post ferric-reducing antioxidant power (FRAP), trolox-equivalent antioxidant capacity (TEAC), uric acid, reduced/oxidised glutathione content (GSH/GSSG) and total peroxidase activity (TPO) were evaluated in whole saliva of both groups. All non-enzymatic antioxidant parameters were augmented by fluoride-gel NaF 2% application, whereas a notable reduction (31%) of peroxidase activity was concomitantly observed in the children's saliva (p ≤ 0.05). Nevertheless, the reducing power of saliva was kept unaltered under these circumstances (p ≤ 0.05). Despite the reduced activity of peroxidase (an important antimicrobial and antioxidant enzyme), the topical fluoride gel NaF 2% favourably stimulated the release of non-enzymatic antioxidant components of saliva, sustaining the reducing power of saliva and the natural defences of the oral cavity. Copyright © 2011 Elsevier Ltd. All rights reserved.

  3. Recent Advances in Enzymatic Fuel Cells: Experiments and Modeling

    Directory of Open Access Journals (Sweden)

    Ivan Ivanov

    2010-04-01

    Full Text Available Enzymatic fuel cells convert the chemical energy of biofuels into electrical energy. Unlike traditional fuel cell types, which are mainly based on metal catalysts, the enzymatic fuel cells employ enzymes as catalysts. This fuel cell type can be used as an implantable power source for a variety of medical devices used in modern medicine to administer drugs, treat ailments and monitor bodily functions. Some advantages in comparison to conventional fuel cells include a simple fuel cell design and lower cost of the main fuel cell components, however they suffer from severe kinetic limitations mainly due to inefficiency in electron transfer between the enzyme and the electrode surface. In this review article, the major research activities concerned with the enzymatic fuel cells (anode and cathode development, system design, modeling by highlighting the current problems (low cell voltage, low current density, stability will be presented.

  4. Enzymatic saccharification of brown seaweed for production of fermentable sugars.

    Science.gov (United States)

    Sharma, Sandeep; Horn, Svein Jarle

    2016-08-01

    This study shows that high drying temperatures negatively affect the enzymatic saccharification yield of the brown seaweed Saccharina latissima. The optimal drying temperature of the seaweed in terms of enzymatic sugar release was found to be 30°C. The enzymatic saccharification process was optimized by investigating factors such as kinetics of sugar release, enzyme dose, solid loading and different blend ratios of cellulases and an alginate lyase. It was found that the seaweed biomass could be efficiently hydrolysed to fermentable sugars using a commercial cellulase cocktail. The inclusion of a mono-component alginate lyase was shown to improve the performance of the enzyme blend, in particular at high solid loadings. At 25% dry matter loading a combined glucose and mannitol concentration of 74g/L was achieved. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. High volumetric power density, non-enzymatic, glucose fuel cells.

    Science.gov (United States)

    Oncescu, Vlad; Erickson, David

    2013-01-01

    The development of new implantable medical devices has been limited in the past by slow advances in lithium battery technology. Non-enzymatic glucose fuel cells are promising replacement candidates for lithium batteries because of good long-term stability and adequate power density. The devices developed to date however use an "oxygen depletion design" whereby the electrodes are stacked on top of each other leading to low volumetric power density and complicated fabrication protocols. Here we have developed a novel single-layer fuel cell with good performance (2 μW cm⁻²) and stability that can be integrated directly as a coating layer on large implantable devices, or stacked to obtain a high volumetric power density (over 16 μW cm⁻³). This represents the first demonstration of a low volume non-enzymatic fuel cell stack with high power density, greatly increasing the range of applications for non-enzymatic glucose fuel cells.

  6. Biocolloids with ordered urease multilayer shells as enzymatic reactors.

    Science.gov (United States)

    Lvov, Y; Caruso, F

    2001-09-01

    The preparation of biocolloids with organized enzyme-containing multilayer shells for exploitation as colloidal enzymatic nanoreactors is described. Urease multilayers were assembled onto submicrometer-sized polystyrene spheres by the sequential adsorption of urease and polyelectrolyte, in a predetermined order, utilizing electrostatic interactions for layer growth. The catalytic activity of the biocolloids increased proportionally with the number of urease layers deposited on the particles, demonstrating that biocolloid particles with tailored enzymatic activities can be produced. It was further found that precoating the latex spheres with nanoparticles (40-nm silica or 12-nm magnetite) enhanced both the stability (with respect to adsorption) and enzymatic activity of the urease multilayers. The presence of the magnetite nanoparticle coating also provided a magnetic function that allowed the biocolloids to be easily and rapidly separated with a permanent magnet. The fabrication of such colloids opens new avenues for the application of bioparticles and represents a promising route for the creation of complex catalytic particles.

  7. Multiple enzymatic profiles of Vibrio parahaemolyticus strains isolated from oysters

    Directory of Open Access Journals (Sweden)

    Renata Albuquerque Costa

    Full Text Available The enzymatic characterization of vibrios has been used as a virulence indicator of sanitary interest. The objective of this study was to determine the enzymatic profile of Vibrio parahaemolyticus strains (n = 70 isolated from Crassostrea rhizophorae oysters. The strains were examined for the presence of gelatinase (GEL, caseinase (CAS, elastase (ELAS, phospholipase (PHOS, lipase (LIP, amilase (AML and DNase. All enzymes, except elastase, were detected in more than 60% of the strains. The most recurrent enzymatic profiles were AML + DNase + PHOS + GEL + LIP (n = 16; 22.9% and AML + CAS + DNase + PHOS + GEL + LIP (n = 21; 30%. Considering the fact that exoenzyme production by vibrios is closely related to virulence, one must be aware of the bacteriological risk posed to human health by the consumption of raw or undercooked oysters.

  8. ENZYMATIC CHANGES IN SNAKE ENVENOMATION- AN OBSERVATIONAL STUDY

    Directory of Open Access Journals (Sweden)

    Sidharth Kapoor

    2017-06-01

    Full Text Available BACKGROUND Snakes are the most feared and the most worshipped living creatures on the earth. Snakes are called venomous when envenomation or human fatalities after their bite are known. Snakebite is an acute medical emergency faced by temperate and tropical regions with heavy rainfall and humid climate. The specific therapy for snakebite in India is still polyvalent ASV and clinical practice ASV is not recommended until the victim of snakebite presents either with the evidence of bite by a poisonous snake such as definite fang marks, swelling or pain at the bite site or with clinical or laboratory evidence of envenomation such as local and systemic bleeding. In some cases, institution of ASV may also be initiated on the identification of offending snake brought by the patient or attendants, but most of these are subjective matters and subject to fallacies. Also, that out of polyvalent and monovalent ASV available, since it is monovalent ASV, which is desirable due to its less side effects and more effectiveness, but its use warrants the identification of snake, which is practically not possible in every case and/or on the objective evidence of peripheral neurological signs and symptoms and haematological alterations, which may not be dependable in many cases. MATERIALS AND METHODS Snake envenomation is in fact a multifactorial stress phenomenon, which produces altered physiological states including death and one of the consequences of the stress phenomenon is generation of several lysosomal enzymes and formation of free radicals. Extensive data search on Medline has failed to show study of this type in any part of the world, so this study being taken up as a preliminary attempt to evaluate the pattern of enzymatic changes in snake envenomation. RESULTS The patients included in the study were be those coming to the Emergency Department of Government Medical College, Jammu, bitten by poisonous snakes during the period May 2003 to April 2004. The

  9. Kinetic study of enzymatic hydrolysis of potato starch

    Directory of Open Access Journals (Sweden)

    Óscar Fernando Castellanos Domínguez

    2004-01-01

    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.

  10. Non-enzymatic U(VI) interactions with biogenic mackinawite

    Science.gov (United States)

    Veeramani, H.; Qafoku, N. P.; Kukkadapu, R. K.; Murayama, M.; Hochella, M. F.

    2011-12-01

    Reductive immobilization of hexavalent uranium [U(VI)] by stimulation of dissimilatory metal and/or sulfate reducing bacteria (DMRB or DSRB) has been extensively researched as a remediation strategy for subsurface U(VI) contamination. These bacteria derive energy by reducing oxidized metals as terminal electron acceptors, often utilizing organic substrates as electron donors. Thus, when evaluating the potential for in-situ uranium remediation in heterogeneous subsurface media, it is important to understand how the presence of alternative electron acceptors such as Fe(III) and sulfate affect U(VI) remediation and the long term behavior and reactivity of reduced uranium. Iron, an abundant subsurface element, represents a substantial sink for electrons from DMRB, and the reduction of Fe(III) leads to the formation of dissolved Fe(II) or to reactive biogenic Fe(II)- and mixed Fe(II)/Fe(III)- mineral phases. Consequently, abiotic U(VI) reduction by reactive forms of biogenic Fe(II) minerals could be a potentially important process for uranium immobilization. In our study, the DMRB Shewanella putrefaciens CN32 was used to synthesize a biogenic Fe(II)-bearing sulfide mineral: mackinawite, that has been characterized by XRD, SEM, HRTEM and Mössbauer spectroscopy. Batch experiments involving treated biogenic mackinawite and uranium (50:1 molar ratio) were carried out at room temperature under strict anoxic conditions. Following complete removal of uranium from solution, the biogenic mackinawite was analyzed by a suite of analytical techniques including XAS, HRTEM and Mössbauer spectroscopy to determine the speciation of uranium and investigate concomitant Fe(II)-phase transformation. Determining the speciation of uranium is critical to success of a remediation strategy. The present work elucidates non-enzymatic/abiotic molecular scale redox interactions between biogenic mackinawite and uranium.

  11. Enzymatic hydrolysis of organic phosphorus in swine manure and soil.

    Science.gov (United States)

    He, Zhongqi; Griffin, Timothy S; Honeycutt, C Wayne

    2004-01-01

    Organic phosphorus (Po) exists in many chemical forms that differ in their susceptibility to hydrolysis and, therefore, bioavailability to plants and microorganisms. Identification and quantification of these forms may significantly contribute to effective agricultural P management. Phosphatases catalyze reactions that release orthophosphate (Pi) from Po compounds. Alkaline phosphatase in tris-HCl buffer (pH 9.0), wheat (Triticum aestivum L.) phytase in potassium acetate buffer (pH 5.0), and nuclease P1 in potassium acetate buffer (pH 5.0) can be used to classify and quantify Po in animal manure. Background error associated with different pH and buffer systems is observed. In this study, we improved the enzymatic hydrolysis approach and tested its applicability for investigating Po in soils, recognizing that soil and manure differ in numerous physicochemical properties. We applied (i) acid phosphatase from potato (Solanum tuberosum L.), (ii) acid phosphatases from both potato and wheat germ, and (iii) both enzymes plus nuclease P1 to identify and quantify simple labile monoester P, phytate (myo-inositol hexakis phosphate)-like P, and DNA-like P, respectively, in a single pH/buffer system (100 mM sodium acetate, pH 5.0). This hydrolysis procedure released Po in sequentially extracted H2O, NaHCO3, and NaOH fractions of swine (Sus scrofa) manure, and of three sandy loam soils. Further refinement of the approach may provide a universal tool for evaluating hydrolyzable Po from a wide range of sources.

  12. Adhesion improvement of lignocellulosic products by enzymatic pre-treatment.

    Science.gov (United States)

    Widsten, Petri; Kandelbauer, Andreas

    2008-01-01

    Enzymatic bonding methods, based on laccase or peroxidase enzymes, for lignocellulosic products such as medium-density fiberboard and particleboard are discussed with reference to the increasing costs of presently used petroleum-based adhesives and the health concerns associated with formaldehyde emissions from current composite products. One approach is to improve the self-bonding properties of the particles by oxidation of their surface lignin before they are fabricated into boards. Another method involves using enzymatically pre-treated lignins as adhesives for boards and laminates. The application of this technology to achieve wet strength characteristics in paper is also reviewed.

  13. Optimization of Substrate Feeding for Enzymatic Biodiesel Production

    DEFF Research Database (Denmark)

    Price, Jason Anthony; Huusom, Jakob Kjøbsted; Nordblad, Mathias

    2013-01-01

    to be effective in mitigating the effects of substrate inhibition. Using enzymatic biodiesel production as a case study, the volumetric productivity of the reactor is increased while minimizing inactivation of the enzyme due to the alcohol. This is done by using a simple optimization routine where the substrate...... (both the vegetable oil and alcohol) feed rate/concentration is manipulated simultaneously. The results of the simulation were tested in the laboratory and are sufficiently positive to suggest the implementation of a feeding strategy for large scale enzymatic biodiesel production...

  14. Chemical and enzymatic stability of amino acid prodrugs containing methoxy, ethoxy and propylene glycol linkers.

    Science.gov (United States)

    Gupta, Deepak; Gupta, Sheeba Varghese; Lee, Kyung-Dall; Amidon, Gordon L

    2009-01-01

    We evaluated the chemical and enzymatic stabilities of prodrugs containing methoxy, ethoxy and propylene glycol linkers in order to find a suitable linker for prodrugs of carboxylic acids with amino acids. l-Valine and l-phenylalanine prodrugs of model compounds (benzoic acid and phenyl acetic acid) containing methoxy, ethoxy and propylene glycol linkers were synthesized. The hydrolysis rate profile of each compound was studied at physiologically relevant pHs (1.2, 4, 6 and 7.4). Enzymatic hydrolysis of propylene glycol containing compounds was studied using Caco-2 homogenate as well as purified enzyme valacyclovirase. It was observed that the stability of the prodrugs increases with the linker length (propyl > ethyl > methyl). The model prodrugs were stable at acidic pH as compared to basic pH. It was observed that the prodrug with the aliphatic amino acid promoiety was more stable compared to its aromatic counterpart. The comparison between benzyl and the phenyl model compounds revealed that the amino acid side chain is significant in determining the stability of the prodrug whereas the benzyl or phenyl carboxylic acid had little or no effect on the stability. The enzymatic activation studies of propylene glycol linker prodrug in the presence of valacyclovirase and cell homogenate showed faster generation of the parent drug at pH 7.4. The half-life of prodrugs at pH 7.4 was more than 12 h, whereas in the presence of cell homogenate the half-lives were less than 1 h. Hydrolysis by Caco-2 homogenate generated the parent compound in two steps, where the prodrug was first converted to the intermediate, propylene glycol benzoate, which was then converted to the parent compound (benzoic acid). Enzymatic hydrolysis of propylene glycol containing prodrugs by valacyclovirase showed hydrolysis of the amino acid ester part to generate the propylene glycol ester of model compound (propylene glycol benzoate) as the major product. The amino acid prodrugs containing methoxy

  15. Enzymatic Synthesis of Esculin Ester in Ionic Liquids Buffered with Organic Solvents

    DEFF Research Database (Denmark)

    Hu, Yifan; Guo, Zheng; Lue, Bena-Marie

    2009-01-01

    The enzymatic esterification of esculin catalyzed by Candida antarctica lipase B (Novozym 435) was carried out in ionic liquid (IL)-organic solvent mixed systems in comparison with individual systems. The reaction behaviors in IL-organic solvents were systemically evaluated using acetone as a model...... in IL-acetone mixtures made it possible to improve the solubility of esculin while the effects of ILs on lipase activity were minimized. Following the benignity of ILs to lipase activity, the anions of ILs were ranked in the order as [Tf2N](-) > [PF6](-) > [BF4](-) > [CF3SO3](-) > [C4F9SO3](-) > [TAF...

  16. Changes in non-enzymatic antioxidant capacity and lipid peroxidation during germination of white, yellow and purple maize seeds

    International Nuclear Information System (INIS)

    Deng, B.; Zhang, Y.; Yang, K.

    2016-01-01

    In this study, the changes in non-enzymatic antioxidant capacity and lipid peroxidation during the germination process of purple, yellow and white maize seeds were compared, under favorable conditions. Results showed that germination can increase non-enzymatic antioxidant capacity (evaluated with ferric reducing power and 2, 2-diphenyl-1-picryl-hydrazyl-hydrate radical scavenging capacity) and lipid peroxidation levels for all these seeds. In addition, non-enzymatic antioxidant capacity observed in the germinating seeds were in the order of purple > yellow > white. However, the highest and lowest levels of lipid peroxidation could be seen during the germination processes of the white and purple seeds, respectively. In addition, the germination rates of the seeds followed the order of white > yellow > purple. Further studies showed that H/sub 2/O/sub 2/ treatment can significantly promote seed germination, especially for purple seeds. In addition, DMTU (dimethylthiourea), a specific scavenger for H/sub 2/O/sub 2/, could slightly but significantly arrest dormancy release. Data analysis showed that a high negative correlation (R/sup 2/ = -0.955) existed between non-enzymatic antioxidant capacity and germination rates. However, a high positive correlation (R/sup 2/ = 0.860) could be detected between lipid peroxidation and germination rates. Finally, lipid peroxidation as a possible novel signaling mechanism for seed germination has been discussed under stress-free conditions. (author)

  17. Enzymatic generation of hydrogen peroxide shows promising antifouling effect

    DEFF Research Database (Denmark)

    Kristensen, J.B.; Olsen, Stefan Møller; Laursen, B.S.

    2010-01-01

    Proteobacteria, tested in microtiter plates. However, enzymatically produced H2O2 released from a coating did not impede biofilm formation by bacteria in natural seawater tested in a biofilm reactor. A field trial revealed a noticeable effect of the enzyme system: after immersion in the North Sea for 97 days...

  18. Reversible sol-gel-sol medium for enzymatic optical biosensors

    NARCIS (Netherlands)

    Safaryan, S.; Yakovlev, A.; Pidko, E.A.; Vinogradov, A.; Vinogradov, V.

    2017-01-01

    In this paper we for the first time report a reversible sol-gel-sol approach to obtain optical enzymatic biosensors with improved enzyme stability and good sensitivity by using desktop inkjet printing. The developed technique is based on the bio-inorganic inks allowing for a sol-gel-sol transition

  19. The properties of main oxidases associated with enzymatic ...

    African Journals Online (AJOL)

    Lenovo User

    2012-07-03

    Jul 3, 2012 ... 1College of Food Science and Engineering, Shandong Agricultural University, Taian, Shandong, 271018,. People's Republic of China. 2The Central Hospital of Taian, Taian, Shandong ... pear vinegar, preserved pear and canned pear. However, enzymatic browning during processing impairs sensory.

  20. Enzymatic Browning in Sugar Beet Leaves (Beta vulgaris L.)

    NARCIS (Netherlands)

    Vissers, Anne; Kiskini, Alexandra; Hilgers, Roelant; Marinea, Marina; Wierenga, Peter Alexander; Gruppen, Harry; Vincken, Jean Paul

    2017-01-01

    Sugar beet (Beta vulgaris L.) leaves of 8 month (8m) plants showed more enzymatic browning than those of 3 month (3m). Total phenolic content increased from 4.6 to 9.4 mg/g FW in 3m and 8m, respectively, quantitated by

  1. Enzymatic epoxidation of biodiesel optimized by response surface ...

    African Journals Online (AJOL)

    During the enzymatic epoxidation of biodiesel, stearic acid was selected as oxygen carrier. Enzyme screening and the load of stearic acid were investigated. The effects of four main reaction conditions including reaction time, temperature, enzyme load, and mole ratio of H2O2/C=C-bonds on the epoxy oxygen group content ...

  2. Enzymatic biodiesel production from sludge palm oil (SPO) using ...

    African Journals Online (AJOL)

    Biodiesel is a non-toxic, renewable and environmental friendly fuel. This study involved the production of biodiesel from sludge palm oil (SPO), a low-cost waste oil via enzymatic catalysis. The enzyme catalyst was a Candida cylindracea lipase, locally-produced using palm oil mill effluent as the low cost based medium.

  3. Short-time ultrasonication treatment in enzymatic hydrolysis of biomass

    Science.gov (United States)

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

    2013-01-01

    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...

  4. Enzymatic modification of phospholipids forfunctional applications and human nutrition

    DEFF Research Database (Denmark)

    Guo, Zheng; Vikbjerg, Anders / Falk; Xu, Xuebing

    2005-01-01

    analogs based on the latest understanding of pivotal role of phospholipids in manifold biological processes, exploration of remarkable application potentials of phospholipids in meliorating human health, as well as development of new chemical and biotechnological approaches applied to the modification...... design. This will of course provide fundamental bases also for the development of enzymatic technology to produce structured or modified phospholipids....

  5. Enzymatic activities of Azotobacter chroococcum and survival in ...

    African Journals Online (AJOL)

    Enzymatic activities of Azotobacter chroococcum and survival in schloropyrifos amended sterile and non-sterile. M Shukla, V Kumar, RL Thakur, N Narula. Abstract. No Abstract. Cameroon Journal of Experimental Biology Vol. 2 (2) 2006: pp. 88-94. AJOL African Journals Online. HOW TO USE AJOL... for Researchers · for ...

  6. ETHANOL ORGANOSOLV PRETREATMENT OF BAMBOO FOR EFFICIENT ENZYMATIC SACCHARIFICATION

    Directory of Open Access Journals (Sweden)

    Zhiqiang Li,

    2012-06-01

    Full Text Available Bamboo is a potential lignocellulosic biomass for the production of bioethanol because of its high cellulose and hemicelluloses content. In this research, ethanol organosolv pretreatment with dilute sulfuric acid as the catalyst was studied in order to enhance enzymatic saccharification of moso bamboo. The addition of 2% (w/w bamboo dilute sulfuric acid in 75% ethanol had a particularly strong effect on fractionation of bamboo. It yielded a solids fraction containing 83.4% cellulose in the treated substrate. The cellulose conversion to glucose yield reached 77.1 to 83.4% after enzymatic hydrolysis of the solids fraction for 48 h at an enzyme loading of 15 FPU cellulase/g cellulose and 30 IU β-glucosidase/g cellulose. The enzymatic hydrolysis rate was significantly accelerated as the ethanol organosolv pretreatment time increased, reaching the highest enzymatic glucose yield of 83.4% after 48 h at 50 °C. The concentrations of fermentation inhibitors such as HMF (5-hydroxy-2-methyl furfural and furfural were 0.96 g/L and 4.38 g/L in the spent liquor after the ethanol organosolv pretreatment, which were slightly lower than the concentrations quantified during H2SO4-water treatment. Spent liquor was diluted with water, and more than 87.2% of lignin in raw bamboo was recovered as ethanol organosolv lignin through the filtration process.

  7. Enzymatic cell disruption of microalgae biomass in biorefinery processes.

    Science.gov (United States)

    Demuez, Marie; Mahdy, Ahmed; Tomás-Pejó, Elia; González-Fernández, Cristina; Ballesteros, Mercedes

    2015-10-01

    When employing biotechnological processes for the procurement of biofuels and bio-products from microalgae, one of the most critical steps affecting economy and yields is the "cell disruption" stage. Currently, enzymatic cell disruption has delivered effective and cost competitive results when compared to mechanical and chemical cell disruption methods. However, the introduction of enzymes implies additional associated cost within the overall process. In order to reduce this cost, autolysis of microalgae is proposed as alternative enzymatic cell disruption method. This review aims to provide the state of the art of enzymatic cell disruption treatments employed in biorefinery processes and highlights the use of endopeptidases. During the enzymatic processes of microalgae life cycle, some lytic enzymes involved in cell division and programmed cell death have been proven useful in performing cell lysis. In this context, the role of endopeptidases is emphasized. Mirroring these natural events, an alternative cell disruption approach is proposed and described with the potential to induce the autolysis process using intrinsic cell enzymes. Integrating induced autolysis within biofuel production processes offers a promising approach to reduce overall global costs and energetic input associated with those of current cell disruption methods. A number of options for further inquiry are also discussed. © 2015 Wiley Periodicals, Inc.

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

    DEFF Research Database (Denmark)

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

    2014-01-01

    .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...

  9. Recombinant EXLX1 from Bacillus subtilis for enhancing enzymatic ...

    African Journals Online (AJOL)

    AJL

    2012-06-21

    Jun 21, 2012 ... enhancing enzymatic hydrolysis of corn stover with low cellulase loadings. Zhang Yan1, He Ming-Xiong2,3*, Wu Bo1, ... University, Chengdu 610064, China. 2Biogas Institute of Ministry of Agriculture, Biomass Energy Technology Research Centre, Section 4-13, Renming Nanlu,. Chengdu 610041, China.

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

    DEFF Research Database (Denmark)

    Prunescu, Remus Mihail; Sin, Gürkan

    2014-01-01

    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...

  11. Enzymatic cell wall degradation of high-pressure-homogenized tomato puree and its effect on lycopene bioaccessibility.

    Science.gov (United States)

    Palmero, Paola; Colle, Ines; Lemmens, Lien; Panozzo, Agnese; Nguyen, Tuyen Thi My; Hendrickx, Marc; Van Loey, Ann

    2016-01-15

    High-pressure homogenization disrupts cell structures, assisting carotenoid release from the matrix and subsequent micellarization. However, lycopene bioaccessibility of tomato puree upon high-pressure homogenization is limited by the formation of a process-induced barrier. In this context, cell wall-degrading enzymes were applied to hydrolyze the formed barrier and enhance lycopene bioaccessibility. The effectiveness of the enzymes in degrading their corresponding substrates was evaluated (consistency, amount of reducing sugars, molar mass distribution and immunolabeling). An in vitro digestion procedure was applied to evaluate the effect of the enzymatic treatments on lycopene bioaccessibility. Enzymatic treatments with pectinases and cellulase were proved to effectively degrade their corresponding cell wall polymers; however, no further significant increase in lycopene bioaccessibility was obtained. A process-induced barrier consisting of cell wall material is not the only factor governing lycopene bioaccessibility upon high-pressure homogenization. © 2015 Society of Chemical Industry.

  12. Pretreatment of sugarcane bagasse using the advanced oxidation process by electron beam for enzymatic hydrolysis of cellulose

    International Nuclear Information System (INIS)

    Ribeiro, Marcia Almeida

    2013-01-01

    The sugar cane bagasse is a renewable energy source and a raw material promise in the biofuel production, once represents about 30% of glucose contained in the plant with the potential to be hydrolyzed and then converted to ethanol. The bagasse is composed of cellulose, straight chain of glucose, of hemicellulose, an amorphous polymer consisting of xylose, arabinose, galactose, and mannose, and of lignin, a complex polymer consisting of fenilpropan units that acts as waterproof coating on the fibers, which is hard to remove due its recalcitrant nature. The aim of this work was to study the electron beam processing as a pretreatment of sugarcane bagasse to enzymatic hydrolysis of cellulose. The pretreatment of sugarcane bagasse is one of the most important steps to make this material economically viable and competitive on the energy production. As a pretreatment the electron beam processing can weak the hemicellulose and lignin structures by the action highly reactive radicals that breaks the links, reducing the degree of polymerization fibers. It was evaluated the chemical and structural modifications on fibers caused by the irradiation, the enzymatic hydrolysis of electron beam as the only pretreatment and combined to steam explosion. For enzymatic hydrolysis it was used the commercial enzymes from Novozymes. The radiation processing promotes changes in structure and composition of sugarcane bagasse, increasing the solubility, that is related to hemicellulose and cellulose cleavage, and also increasing the enzymatic conversion yield. In the case of exploded bagasse there is no changes in the enzymatic hydrolysis yield, however the electron beam processing promoted a 67% reduction of furfural, that is formed in the steam explosion process. (author)

  13. Enzymatic detection of formalin-fixed museum specimens for DNA analysis and enzymatic maceration of formalin-fixed specimens

    DEFF Research Database (Denmark)

    Sørensen, Margrethe; Redsted Rasmussen, Arne; Simonsen, Kim Pilkjær

    2016-01-01

    % ethanol. The method was subsequently tested on wild-living preserved specimens and an archived specimen. The protease enzyme used was SavinaseH 16 L, Type EX from Novozymes A/S. The enzymatic screening test demands only simple laboratory equipment. The method is useful for natural history collections...

  14. ENZYMATIC KINETIC STUDY HYDROLASE FROM CITRUS

    Directory of Open Access Journals (Sweden)

    Israel Hernández

    2015-09-01

    Full Text Available In this paper the degrading activity of enzymes derived from orange peels (Citrus x sinensis, grapefruit (Citrus paradise and pineapple (Ananas comosus on the organic matter in wastewater is evaluated. This activity is measured indirectly by quantifying the biochemical oxygen demand (COD before and after degradation process based on a period of time using the HACH DR / 2010, and then the kinetic study was performed by the differential method and integral with the experimental data, obtaining a reaction order of 1 to pectinase (orange, and order 2 for bromelain (pineapple.

  15. Non-enzymatic glucose sensing platform using self assembled cobalt oxide/graphene nanocomposites immobilized graphite modified electrode

    DEFF Research Database (Denmark)

    Vivekananth, R.; Babu, R. Suresh; Prasanna, K.

    2018-01-01

    A new strategy to prepare the densely packed cobalt oxide (Co3O4)/graphene nanocomposites by a self-assembly method were adopted in this work. A new non-enzymatic glucose determination has been fabricated by using Co3O4/graphene nanocomposites modified electrode as a sensing material. The nanocom...... of the modified electrode for glucose determination has been evaluated in urine samples....

  16. Fat content affects bioaccessibility and efficiency of enzymatic hydrolysis of lutein esters added to milk and yogurt

    OpenAIRE

    Xavier, Ana Augusta Odorissi; Mercadante, Adriana Zerlotti; Garrido-Fernández, Juan; Pérez-Gálvez, Antonio

    2014-01-01

    © 2014 Elsevier Ltd. Addition of lutein to dairy products is an alternative that widens the range of foods which could be lutein sources. However, bioaccessibility is an essential aspect to be considered during the development of products with added bioactive substances. We evaluated the in vitro bioaccessibility of lutein esters added to milk and yogurt with different fat contents, and determined the efficiency of enzymatic hydrolysis of the esters during digestion. Bioaccessibility of lutei...

  17. Oxidant and enzymatic antioxidant status (gene expression and activity) in the brain of chickens with cold-induced pulmonary hypertension

    Science.gov (United States)

    Hassanpour, Hossein; Khalaji-Pirbalouty, Valiallah; Nasiri, Leila; Mohebbi, Abdonnaser; Bahadoran, Shahab

    2015-11-01

    To evaluate oxidant and antioxidant status of the brain (hindbrain, midbrain, and forebrain) in chickens with cold-induced pulmonary hypertension, the measurements of lipid peroxidation, protein oxidation, antioxidant capacity, enzymatic activity, and gene expression (for catalase, glutathione peroxidase, and superoxide dismutases) were done. There were high lipid peroxidation/protein oxidation and low antioxidant capacity in the hindbrain of cold-induced pulmonary hypertensive chickens compared to control ( P pulmonary hypertension.

  18. Improvement Enzymatic Hydrolysis of Wheat Straw for Bioethanol Production by Combined Treatment of Radiation and Acid

    International Nuclear Information System (INIS)

    Hong, Sung Hyun; Lee, Seung Sik; Bai, Hyoung Woo; Chung, Byung Yeoup

    2012-01-01

    The cost of ethanol production from starch and sucrose for use as a vehicle fuel is ultimately high. Consequently, it has been suggested that the large-scale use of ethanol as a fuel will require the utilization of cellulosic feedstock. Lignocellulosic biomass has the potential to serve as a low cost and renewable feedstock for bioconversion into fermentable sugars, which can be further utilized for biofuel production. It is estimated that there is over one billion tons of biomass available for conversion into biofuels on a renewable basis to displace a substantial portion of the fossil fuels currently consumed within the transportation sector. Among different pretreatment methods such as biological, physical, chemical, and physic-chemical pretreatments, chemical pretreatment using dilute acid as catalyst, which has been extensively evaluated for treating a variety of lignocellulosic feedstocks, is reported as one of the leading pretreatment technologies. Ionizing radiation can easily penetrate lignocellulosic structure and undoubtedly produce free radicals useful in modification of lignin structure as well as breakdown cellulose crystal regions. Phenoxy radicals appeared to be important radical intermediates that ultimately transformed into o-quinonoid structures in lignin. Therefore, ionizing radiation such as gamma ray and electron beam can be a great alternative. In this study, the effect of ionizing irradiation of wheat straw prior to dilute sulfuric acid treatment is investigated. The combined pretreatment for wheat straw was performed to evaluate the efficiency of enzymatic hydrolysis and compared with that of the effect of enzymatic hydrolysis by individual pretreatment

  19. Analytical performances of a new enzymatic assay for hemoglobin A1c.

    Science.gov (United States)

    Jaisson, Stéphane; Desmons, Aurore; Renard, Benoît; Chevelle, Benjamin; Leroy, Nathalie; Gillery, Philippe

    2014-07-01

    HbA1c is considered the gold standard for the follow-up of diabetic patients and a new diagnostic tool for diabetes mellitus, which implies the availability of reliable assay methods. We have evaluated a new assay developed by Abbott Laboratories, based on the enzymatic quantification of HbA1c by a fructosyl dipeptide oxidase using Architect analyzers. Precision, linearity, correlation with a HPLC method, accuracy and potential impact interferences on HbA1c measurement have been evaluated. Intra-day and between-day CVs were lower than 1.2% and linearity was excellent from 19 mmol/mol (3.9%) to 163 mmol/mol (17.1%). The results were well correlated with those obtained by the HPLC (Variant II device, kit NU - BioRad): HbA1c [Architect, mmol/mol]=0.986×HbA1c [Variant II, mmol/mol]+0.713 (r=0.998, n=109). This method provided consistent results with IFCC titrated quality control samples. Classical interferences in HbA1c assays (i.e. labile HbA1c, carbamylated hemoglobin, triglycerides or bilirubin) did not have an impact on HbA1c quantification by this method. This new enzymatic assay proved to be a robust and reliable method for HbA1c measurement suitable for routine practice in clinical chemistry laboratories. Copyright © 2014 Elsevier B.V. All rights reserved.

  20. Enzymatic Hydrolysis Does Not Reduce the Biological Reactivity of Soybean Proteins for All Allergic Subjects.

    Science.gov (United States)

    Panda, Rakhi; Tetteh, Afua O; Pramod, Siddanakoppalu N; Goodman, Richard E

    2015-11-04

    Many soybean protein products are processed by enzymatic hydrolysis to attain desirable functional food properties or in some cases to reduce allergenicity. However, few studies have investigated the effects of enzymatic hydrolysis on the allergenicity of soybean products. In this study the allergenicity of soybean protein isolates (SPI) hydrolyzed by Alcalase, trypsin, chymotrypsin, bromelain, or papain was evaluated by IgE immunoblots using eight soybean-allergic patient sera. The biological relevance of IgE binding was evaluated by a functional assay using a humanized rat basophilic leukemia (hRBL) cell line and serum from one subject. Results indicated that hydrolysis of SPI by the enzymes did not reduce the allergenicity, and hydrolysis by chymotrypsin or bromelain has the potential to increase the allergenicity of SPI. Two-dimensional (2D) immunoblot and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis of the chymotrypsin-hydrolyzed samples indicated fragments of β-conglycinin protein are responsible for the apparent higher allergenic potential of digested SPI.

  1. Respiration and enzymatic activities as indicators of stabilization of sewage sludge composting.

    Science.gov (United States)

    Nikaeen, Mahnaz; Nafez, Amir Hossein; Bina, Bijan; Nabavi, BiBi Fatemeh; Hassanzadeh, Akbar

    2015-05-01

    The objective of this work was to study the evolution of physico-chemical and microbial parameters in the composting process of sewage sludge (SS) with pruning wastes (PW) in order to compare these parameters with respect to their applicability in the evaluation of organic matter (OM) stabilization. To evaluate the composting process and organic matter stability, different microbial activities were compared during composting of anaerobically digested SS with two volumetric ratios, 1:1 and 3:1 of PW:SS and two aeration techniques including aerated static piles (ASP) and turned windrows (TW). Dehydrogenase activity, fluorescein diacetate hydrolysis, and specific oxygen uptake rate (SOUR) were used as microbial activity indices. These indices were compared with traditional parameters, including temperature, pH, moisture content, organic matter, and C/N ratio. The results showed that the TW method and 3:1 (PW:SS) proportion was superior to the ASP method and 1:1 proportion, since the former accelerate the composting process by catalyzing the OM stabilization. Enzymatic activities and SOUR, which reflect microbial activity, correlated well with temperature fluctuations. Based on these results it appears that SOUR and the enzymatic activities are useful parameters to monitor the stabilization of SS compost. Copyright © 2015 Elsevier Ltd. All rights reserved.

  2. Fish protein hydrolysate production from sardine solid waste by crude pepsin enzymatic hydrolysis in a bioreactor coupled to an ultrafiltration unit

    International Nuclear Information System (INIS)

    Benhabiles, M.S.; Abdi, N.; Drouiche, N.; Lounici, H.; Pauss, A.; Goosen, M.F.A.; Mameri, N.

    2012-01-01

    The aims of the study were to optimize the production a fish protein hydrolysate (FPH) by enzymatic hydrolysis of sardine solid waste using crude pepsin, and to scale up the process in a bioreactor coupled to an ultrafiltration unit for product recovery. Results showed that the crude pepsin prepared by autolysis of the mucous membranes of a sheep stomach at optimal conditions (i. e. pH = 1.5–2 and incubation time of 6 h) could be satisfactory used for the enzymatic hydrolysis of fish solid waste. The optimal conditions for enzymatic reaction were: temperature 48 °C, and pH 1.5. The scale up of the enzymatic hydrolysis and the coupling of the reactor an ultrafiltration unit to concentrate the hydrolysate gave good results with a rejection coefficient for the protein hydrolysate product in the range of 90%. The volumetric concentration factor was 2.5, with a permeate flux of 200 L m −2 bar −1 . However, the results also suggest that the ultrafiltration product concentration process may be operating beyond the critical flux at which point irreversible membrane fouling occurs. - Highlights: ► Evaluating to produce a (FPH) by enzymatic hydrolysis of sardine solid wastes was achieved. ► Investigation of key parameters for optimal conditions for enzymatic hydrolysis have been studied. ► Valorization of sardine waste was realized by enzymatic hydrolysis process. ► Performances of this enzyme gave comparable results to those obtained with commercial pepsin. ► The nutritional quality of the FPH produced appears to be satisfactory.

  3. Fish protein hydrolysate production from sardine solid waste by crude pepsin enzymatic hydrolysis in a bioreactor coupled to an ultrafiltration unit

    Energy Technology Data Exchange (ETDEWEB)

    Benhabiles, M.S.; Abdi, N. [National Polytechnic school of Algiers, B.P. 182-16200, El Harrach, Algiers (Algeria); Drouiche, N., E-mail: nadjibdrouiche@yahoo.fr [National Polytechnic school of Algiers, B.P. 182-16200, El Harrach, Algiers (Algeria); Silicon Technology Development Unit (UDTS) 2, Bd Frantz Fanon BP140, Alger-7 Merveilles, 16000 (Algeria); Lounici, H. [National Polytechnic school of Algiers, B.P. 182-16200, El Harrach, Algiers (Algeria); Pauss, A. [University of Technology of Compiegne, Departement Genie chimique,B.P. 20.509, 60205 Compiegne cedex (France); Goosen, M.F.A. [Alfaisal University, Riyadh (Saudi Arabia); Mameri, N. [University of Technology of Compiegne, Departement Genie chimique,B.P. 20.509, 60205 Compiegne cedex (France)

    2012-05-01

    The aims of the study were to optimize the production a fish protein hydrolysate (FPH) by enzymatic hydrolysis of sardine solid waste using crude pepsin, and to scale up the process in a bioreactor coupled to an ultrafiltration unit for product recovery. Results showed that the crude pepsin prepared by autolysis of the mucous membranes of a sheep stomach at optimal conditions (i. e. pH = 1.5-2 and incubation time of 6 h) could be satisfactory used for the enzymatic hydrolysis of fish solid waste. The optimal conditions for enzymatic reaction were: temperature 48 Degree-Sign C, and pH 1.5. The scale up of the enzymatic hydrolysis and the coupling of the reactor an ultrafiltration unit to concentrate the hydrolysate gave good results with a rejection coefficient for the protein hydrolysate product in the range of 90%. The volumetric concentration factor was 2.5, with a permeate flux of 200 L m{sup -2} bar{sup -1}. However, the results also suggest that the ultrafiltration product concentration process may be operating beyond the critical flux at which point irreversible membrane fouling occurs. - Highlights: Black-Right-Pointing-Pointer Evaluating to produce a (FPH) by enzymatic hydrolysis of sardine solid wastes was achieved. Black-Right-Pointing-Pointer Investigation of key parameters for optimal conditions for enzymatic hydrolysis have been studied. Black-Right-Pointing-Pointer Valorization of sardine waste was realized by enzymatic hydrolysis process. Black-Right-Pointing-Pointer Performances of this enzyme gave comparable results to those obtained with commercial pepsin. Black-Right-Pointing-Pointer The nutritional quality of the FPH produced appears to be satisfactory.

  4. Enzymatic production of ceramide from sphingomyelin

    DEFF Research Database (Denmark)

    Zhang, Long; Hellgren, Lars; Xu, Xuebing

    2006-01-01

    -saturated organic solvent) system. Among the screened phospholipase C, the Clostridium petfringens enzyme had the highest sphingomyetin conversion rate, with very small temperature dependence. Addition of ethanol to the system markedly enhanced the rate of ceramide formation, and a mixture of ethyl acetate: hexane...... (50:50) was the best organic solvent tested. Other factors such as (NH4)(2)SO4, MCI and CaCl, were also tested but excluded for further consideration. On the basis of the initial experiments, the reaction system was optimized using response surface methodology including five factors (enzyme amount...... systern evaluation and optimization, with the optimal conditions at 75 min reaction time, 3 U ml(-1) enzyme amount, 6% water amount, 1.8% ethanol arnount and 46% hexane in ethylacetate. (c) 2005 Elsevier B.V. All rights reserved....

  5. Enzymatic spectrophotometric reaction rate determination of aspartame

    Directory of Open Access Journals (Sweden)

    Trifković Kata T.

    2015-01-01

    Full Text Available Aspartame is an artificial sweetener of low caloric value (approximately 200 times sweeter than sucrose. Aspartame is currently permitted for use in food and beverage production in more than 90 countries. The application of aspartame in food products requires development of rapid, inexpensive and accurate method for its determination. The new assay for determination of aspartame was based on set of reactions that are catalyzed by three different enzymes: α-chymotrypsin, alcohol oxidase and horseradish peroxidase. Optimization of the proposed method was carried out for: (i α-chymotrypsin activity; (ii time allowed for α-chymotrypsin action, (iii temperature. Evaluation of the developed method was done by determining aspartame content in “diet” drinks, as well as in artificial sweetener pills. [Projekat Ministarstva nauke Republike Srbije, br. III46010

  6. Improving chocolate flavor in poor-quality cocoa almonds by enzymatic treatment.

    Science.gov (United States)

    Oliveira, Hilana Salete Silva; Mamede, Maria Eugênia Oliveira; Góes-Neto, Aristóteles; Koblitz, Maria Gabriela Bello

    2011-01-01

    This paper proposes a method to enzymatically treat poor-quality cocoa almonds (known as "slate") to ensure the formation of chocolate flavor precursors. The production of flavor precursors improves the quality of these almonds, which are usually responsible for the low quality of the liquor produced. Proteases and carboxypeptidases from different sources were tested under various conditions. The different treatments were evaluated by chemical analysis (hydrolysis efficiency) and sensory analysis of the treated material compared to good-quality cocoa almonds. The results show that it is possible, through the use of microbial enzymes, to generate the mixture of compounds that will release, after roasting, the characteristic chocolate flavor in poor-quality almonds. However, it is necessary to optimize the conditions of enzymatic treatment to obtain better results and thus establish a process that can be used for industrial purposes for manufacturing cocoa and chocolate. The basidiomycete Moniliophtora perniciosa is the causative agent of witches' broom disease (WBD) of the cocoa tree, whose seeds are the source of chocolate. It is the most important phytopathological problem of cocoa-producing areas of the American continent, and has decimated the Brazilian cocoa industry. In Bahia (Brazil), M. perniciosa was identified in 1989 and, as a consequence of its spreading, the annual production of cocoa almonds dropped from 450,000 to 90,000 tons within 12 y, reducing export values from an all-time high of about US$ 1 billion to 110 million. The high incidence of WBD incapacitates Brazil to produce enough cocoa almonds even for the internal market, leading the country to import low-quality cocoa almonds mainly from African countries. Our work proposes an enzymatic treatment to increase the quality of that cocoa almonds and, consequently, to improve the quality of the chocolate produced and consumed in the country. © 2011 Institute of Food Technologists®

  7. Enzymatic depolymerization of gum tragacanth: bifidogenic potential of low molecular weight oligosaccharides.

    Science.gov (United States)

    Gavlighi, Hassan Ahmadi; Michalak, Malwina; Meyer, Anne S; Mikkelsen, J Dalgaard

    2013-02-13

    Gum tragacanth derived from the plant "goat's horn" (Astragalus sp.) has a long history of use as a stabilizing, viscosity-enhancing agent in food emulsions. The gum contains pectinaceous arabinogalactans and fucose-substituted xylogalacturonans. In this work, gum tragacanth from Astragalus gossypinus was enzymatically depolymerized using Aspergillus niger pectinases (Pectinex BE Color). The enzymatically degraded products were divided into three molecular weight fractions via membrane separation: HAG1 10 kDa. Compositional and linkage analyses showed that these three fractions also varied with respect to composition and structural elements: HAG1 and HAG2 were enriched in arabinose, galactose, and galacturonic acid, but low in fucose and xylose, whereas HAG3 was high in (terminal) xylose, fucose, and 1,4-bonded galacturonic acid, but low in arabinose and galactose content. The growth-stimulating potential of the three enzymatically produced gum tragacanth fractions was evaluated via growth assessment on seven different probiotic strains in single-culture fermentations on Bifidobacterium longum subsp. longum (two strains), B. longum subsp. infantis (three strains), Lactobacillus acidophilus , B. lactis, and on one pathogenic strain of Clostridium perfringens . The fractions HAG1 and HAG2 consistently promoted higher growth of the probiotic strains than HAG3, especially of the three B. longum subsp. infantis strains, and the growth promotion on HAG1 and HAG2 was better than that on galactan (control). HAG3 completely inhibited the growth of the C. perfringens strain. Tragacanth gum is thus a potential source of prebiotic carbohydrates that exert no viscosity effects and which may find use as natural functional food ingredients.

  8. The dual effects of Maillard reaction and enzymatic hydrolysis on the antioxidant activity of milk proteins.

    Science.gov (United States)

    Oh, N S; Lee, H A; Lee, J Y; Joung, J Y; Lee, K B; Kim, Y; Lee, K W; Kim, S H

    2013-08-01

    The objective of this study was to determine the enhanced effects on the biological characteristics and antioxidant activity of milk proteins by the combination of the Maillard reaction and enzymatic hydrolysis. Maillard reaction products were obtained from milk protein preparations, such as whey protein concentrates and sodium caseinate with lactose, by heating at 55°C for 7 d in sodium phosphate buffer (pH 7.4). The Maillard reaction products, along with untreated milk proteins as controls, were hydrolyzed for 0 to 3h with commercial proteases Alcalase, Neutrase, Protamex, and Flavorzyme (Novozymes, Bagsværd, Denmark). The antioxidant activity of hydrolyzed Maillard reaction products was determined by reaction with 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt, their 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity, and the ability to reduce ferric ions. Further characteristics were evaluated by the o-phthaldialdehyde method and sodium dodecyl sulfate-PAGE. The degree of hydrolysis gradually increased in a time-dependent manner, with the Alcalase-treated Maillard reaction products being the most highly hydrolyzed. Radical scavenging activities and reducing ability of hydrolyzed Maillard reaction products increased with increasing hydrolysis time. The combined products of enzymatic hydrolysis and Maillard reaction showed significantly greater antioxidant activity than did hydrolysates or Maillard reaction products alone. The hydrolyzed Maillard reaction products generated by Alcalase showed significantly higher antioxidant activity when compared with the other protease products and the antioxidant activity was higher for the whey protein concentrate groups than for the sodium caseinate groups. These findings indicate that Maillard reaction products, coupled with enzymatic hydrolysis, could act as potential antioxidants in the pharmaceutical, food, and dairy industries. Copyright © 2013 American Dairy Science Association

  9. Enzymatic Extraction of Hydroxycinnamic Acids from Coffee Pulp

    Directory of Open Access Journals (Sweden)

    Ernesto Favela-Torres

    2011-01-01

    Full Text Available Ferulic, caffeic, p-coumaric and chlorogenic acids are classified as hydroxycinnamic acids, presenting anticarcinogenic, anti-inflammatory and antioxidant properties. In this work, enzymatic extraction has been studied in order to extract high value-added products like hydroxycinnamic acids from coffee pulp. A commercial pectinase and enzyme extract produced by Rhizomucor pusillus strain 23aIV in solid-state fermentation using olive oil or coffee pulp (CP as an inducer of the feruloyl esterase activity were evaluated separately and mixed. The total content (covalently linked and free of ferulic, caffeic, p-coumaric and chlorogenic acids was 5276 mg per kg of coffee pulp. Distribution was as follows (in %: chlorogenic acid 58.7, caffeic acid 37.6, ferulic acid 2.1 and p-coumaric acid 1.5. Most of the hydroxycinnamic acids were covalently bound to the cell wall (in %: p-coumaric acid 97.2, caffeic acid 94.4, chlorogenic acid 76.9 and ferulic acid 73.4. The content of covalently linked hydroxycinnamic acid was used to calculate the enzyme extraction yield. The maximum carbon dioxide rate for the solid-state fermentation using olive oil as an inducer was higher and it was reached in a short cultivation time. Nevertheless, the feruloyl esterase (FAE activity (units per mg of protein obtained in the fermentation using CP as an inducer was 31.8 % higher in comparison with that obtained in the fermentation using olive oil as the inducer. To our knowledge, this is the first report indicating the composition of both esterified and free ferulic, caffeic, p-coumaric and chlorogenic acids in coffee pulp. The highest yield of extraction of hydroxycinnamic acids was obtained by mixing the produced enzyme extract using coffee pulp as an inducer and a commercial pectinase. Extraction yields were as follows (in %: chlorogenic acid 54.4, ferulic acid 19.8, p-coumaric acid 7.2 and caffeic acid 2.3. An important increase in the added value of coffee pulp was mainly

  10. Enzymatic aqueous technology for simultaneous coconut protein and oil extraction

    Directory of Open Access Journals (Sweden)

    Coelho, Maria A. Z.

    2003-03-01

    Full Text Available In this study, the following commercial enzymes were evaluated in the enzymatic extraction of oil and protein from coconut: Celluclast, Termamyl, Viscozyme, Neutrase y Protease. Preliminary experiments were carried out for selection of enzymes, enzyme concentration and incubation time. In order to verify the effects of pH and water/substrate ratio, a response surface methodology was applied based in a 32 full factorial experimental design. The pH is the most meaningful parameter on oil and protein extraction yields, with a significance level higher than 90%. The maximal extraction yields of oil and protein emulsion (83% was reached using Viscozyme L and subsequently Neutrase 1.5 MG at concentrations of 0.6% (w/w and 0.3% (w/w, respectively, total incubation time of 60 minutes, substrate/water ratio 1:6 and pH around 7 (no adjustment.En este trabajo diferentes enzimas hidroliticos fueron usados para la extracción de aceite y proteína de coco: Celluclast, Termamyl, Viscozyme, Neutrase y Protease producidos por Novo Nordisk. Experimientos preliminares fueron encaminados a la selección de los extractos enzimaticos, de las concentraciones de las enzimas y de los tiempos de incubación. La técnica de análisis de varianza, mediante un delineamiento factorial en tres niveles, fue usada para la optimización de los procesos con relación a los parámetros pH y razón sustrato/agua. Comparando con la extracción acuosa todos los enzimas hidroliticos incrementan la extracción de aceite y proteína vegetal de coco. Los coeficientes de correlación muestran que el pH fué el parámetro mas significativo (pViscozyme y Neutrase 0.6 % (w/w y 0.3 % (w/w, respectivamente, tiempo de incubación 30 minutos, razón sustrato/agua 1:6 y pH 7.

  11. Amyloglucosidase enzymatic reactivity inside lipid vesicles

    Directory of Open Access Journals (Sweden)

    Kim Jin-Woo

    2007-10-01

    Full Text Available Abstract Efficient functioning of enzymes inside liposomes would open new avenues for applications in biocatalysis and bioanalytical tools. In this study, the entrapment of amyloglucosidase (AMG (EC 3.2.1.3 from Aspergillus niger into dipalmitoylphosphatidylcholine (DPPC multilamellar vesicles (MLVs and large unilamellar vesicles (LUVs was investigated. Negative-stain, freeze-fracture, and cryo-transmission electron microscopy images verified vesicle formation in the presence of AMG. Vesicles with entrapped AMG were isolated from the solution by centrifugation, and vesicle lamellarity was identified using fluorescence laser confocal microscopy. The kinetics of starch hydrolysis by AMG was modeled for two different systems, free enzyme in aqueous solution and entrapped enzyme within vesicles in aqueous suspension. For the free enzyme system, intrinsic kinetics were described by a Michaelis-Menten kinetic model with product inhibition. The kinetic constants, Vmax and Km, were determined by initial velocity measurements, and Ki was obtained by fitting the model to experimental data of glucose concentration-time curves. Predicted concentration-time curves using these kinetic constants were in good agreement with experimental measurements. In the case of the vesicles, the time-dependence of product (glucose formation was experimentally determined and simulated by considering the kinetic behavior of the enzyme and the permeation of substrate into the vesicle. Experimental results demonstrated that entrapped enzymes were much more stable than free enyzme. The entrapped enzyme could be recycled with retention of 60% activity after 3 cycles. These methodologies can be useful in evaluating other liposomal catalysis operations.

  12. Influência das variáveis de processo na alcoólise enzimática de óleo de mamona Evaluation of the influence of process variables on enzimatic alcoholysis of castor oil

    Directory of Open Access Journals (Sweden)

    D. Oliveira

    2004-06-01

    intermediates to produce a number of oleochemicals and fuel alternatives (biodiesel. In this work, enzymatic alcoholysis of castor bean oil with an immobilized lipase (Lipozyme IM was accomplished using n-hexane as solvent. In order to maximize the esters production a Taguchi design was adopted varying temperature, water and enzyme concentrations in the reaction media and the oil:ethanol molar ratio. The reactions were monitorated by determining the glycerol production. An empirical model was built to evaluate the influence of the process variables on the conversion and afterwards the operating conditions that maximize esters production were established.

  13. Bio-based alkyds by direct enzymatic bulk polymerization

    DEFF Research Database (Denmark)

    Nguyen, Hiep Dinh

    to a corresponding classical reference. In a further development of the system, it has been found possible to use the esters of pentaerythritol and stearic acid in combination with the penta-aze derivative for the preparation of pseudo alkyds containing only pentaerythritol as polyol with high degree of branching....... Bio-based alkyds prepared from a combination of glycerol, and tall oil fatty acids, and azelaic acid by enzymatic polymerization show improved hydrophobicity and lower glass transition temperatures compared to an alkyd prepared from the same raw materials by a classical boiling method. The enzymatic...... of pentaerythritol derivatized with azelaic acid (or penta-aze) was examined and tested for the production of more branched alkyd systems. A photostability test validated the concept, and the method also resulted in alkyds with improved hydrophobicity and lower glass transition temperatures compared...

  14. Perspectives for the industrial enzymatic production of glycosides.

    Science.gov (United States)

    de Roode, B Mattheus; Franssen, Maurice C R; van der Padt, Albert; Boom, Remko M

    2003-01-01

    Glycosides are of commercial interest for industry in general and specifically for the pharmaceutical and food industry. Currently chemical preparation of glycosides will not meet EC food regulations, and therefore chemical preparation of glycosides is not applicable in the food industry. Thus, enzyme-catalyzed reactions are a good alternative. However, until now the low yields obtained by enzymatic methods prevent the production of glycosides on a commercial scale. Therefore, high yields should be established by a combination of optimum reaction conditions and continuous removal of the product. Unfortunately, a bioreactor for the commercial scale production of glycosides is not available. The aim of this article is to discuss the literature with respect to enzymatic production of glycosides and the design of an industrially viable bioreactor system.

  15. EFFECT OF LIGNIN CONTENT ON ENZYMATIC HYDROLYSIS OF FURFURAL RESIDUES

    Directory of Open Access Journals (Sweden)

    Jianxin Jiang

    2011-02-01

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

  16. Enhanced enzymatic cellulose degradation by cellobiohydrolases via product removal

    DEFF Research Database (Denmark)

    Ahmadi Gavlighi, Hassan; Meyer, Anne S.; Mikkelsen, Jørn Dalgaard

    2013-01-01

    Product inhibition by cellobiose decreases the rate of enzymatic cellulose degradation. The optimal reaction conditions for two Emericella (Aspergillus) nidulans-derived cellobiohydrolases I and II produced in Pichia pastoris were identified as CBHI: 52 °C, pH 4.5–6.5, and CBHII: 46 °C, pH 4.......8. The optimum in a mixture of the two was 50 °C, pH 4.9. An almost fourfold increase in enzymatic hydrolysis yield was achieved with intermittent product removal of cellobiose with membrane filtration (2 kDa cut-off): The conversion of cotton cellulose after 72 h was ~19 % by weight, whereas the conversion...

  17. Enzymatic biodiesel synthesis. Key factors affecting efficiency of the process

    Energy Technology Data Exchange (ETDEWEB)

    Szczesna Antczak, Miroslawa; Kubiak, Aneta; Antczak, Tadeusz; Bielecki, Stanislaw [Institute of Technical Biochemistry, Faculty of Biotechnology and Food Sciences, Technical University of Lodz, Stefanowskiego 4/10, 90-924 Lodz (Poland)

    2009-05-15

    Chemical processes of biodiesel production are energy-consuming and generate undesirable by-products such as soaps and polymeric pigments that retard separation of pure methyl or ethyl esters of fatty acids from glycerol and di- and monoacylglycerols. Enzymatic, lipase-catalyzed biodiesel synthesis has no such drawbacks. Comprehension of the latter process and an appreciable progress in production of robust preparations of lipases may soon result in the replacement of chemical catalysts with enzymes in biodiesel synthesis. Engineering of enzymatic biodiesel synthesis processes requires optimization of such factors as: molar ratio of substrates (triacylglycerols: alcohol), temperature, type of organic solvent (if any) and water activity. All of them are correlated with properties of lipase preparation. This paper reports on the interplay between the crucial parameters of the lipase-catalyzed reactions carried out in non-aqueous systems and the yield of biodiesel synthesis. (author)

  18. Quantifying the limits of transition state theory in enzymatic catalysis.

    Science.gov (United States)

    Zinovjev, Kirill; Tuñón, Iñaki

    2017-11-21

    While being one of the most popular reaction rate theories, the applicability of transition state theory to the study of enzymatic reactions has been often challenged. The complex dynamic nature of the protein environment raised the question about the validity of the nonrecrossing hypothesis, a cornerstone in this theory. We present a computational strategy to quantify the error associated to transition state theory from the number of recrossings observed at the equicommittor, which is the best possible dividing surface. Application of a direct multidimensional transition state optimization to the hydride transfer step in human dihydrofolate reductase shows that both the participation of the protein degrees of freedom in the reaction coordinate and the error associated to the nonrecrossing hypothesis are small. Thus, the use of transition state theory, even with simplified reaction coordinates, provides a good theoretical framework for the study of enzymatic catalysis. Copyright © 2017 the Author(s). Published by PNAS.

  19. Enzymatic hydrolysis and fermentation of agricultural residues to ethanol

    Energy Technology Data Exchange (ETDEWEB)

    Mes-Hartree, M.; Hogan, C.M.; Saddler, J.N.

    1984-01-01

    A combined enzymatic hydrolysis and fermentation process was used to convert steam-treated wheat and barley straw to ethanol. Maximum conversion efficiencies were obtained when the substrates were steamed for 90 s. These substrates could yield over 0.4 g ethanol/g cellulose following a combined enzymatic hydrolysis and fermentation process procedure using culture filtrates derived from Trichoderma harzianum E58. When culture filtrates from Trichoderma reesei C30 and T. reesei QM9414 were used, the ethanol yields obtained were 0.32 and 0.12 g ethanol/g cellulose utilized, respectively. The lower ethanol yields obtained with these strains were attributed to the lower amounts of ..beta..-glucosidase detected in the T. reesei culture filtrates.

  20. On-chip enzymatic microbiofuel cell-powered integrated circuits.

    Science.gov (United States)

    Mark, Andrew G; Suraniti, Emmanuel; Roche, Jérôme; Richter, Harald; Kuhn, Alexander; Mano, Nicolas; Fischer, Peer

    2017-05-16

    A variety of diagnostic and therapeutic medical technologies rely on long term implantation of an electronic device to monitor or regulate a patient's condition. One proposed approach to powering these devices is to use a biofuel cell to convert the chemical energy from blood nutrients into electrical current to supply the electronics. We present here an enzymatic microbiofuel cell whose electrodes are directly integrated into a digital electronic circuit. Glucose oxidizing and oxygen reducing enzymes are immobilized on microelectrodes of an application specific integrated circuit (ASIC) using redox hydrogels to produce an enzymatic biofuel cell, capable of harvesting electrical power from just a single droplet of 5 mM glucose solution. Optimisation of the fuel cell voltage and power to match the requirements of the electronics allow self-powered operation of the on-board digital circuitry. This study represents a step towards implantable self-powered electronic devices that gather their energy from physiological fluids.

  1. Enzymatic reduction of U(VI) in groundwaters

    International Nuclear Information System (INIS)

    Addelouas, A.; Gong, W.; Lutze, W.; Nuttall, E.; Fritz, B.; Crovisier, J.L.

    1999-01-01

    The use of enzymatic reduction of U(VI) in remediation of groundwater contaminated with U(VI) is receiving considerable attention. Certain strains of bacteria can combine the oxidation of an organic compound to the reduction of U(VI) to U(IV), which precipitates as uraninite. In the present study, we tested the reduction of U(VI) in groundwaters with various origins and compositions. In all groundwaters u(VI) was reduced by sulfate reducing bacteria that had been activated by ethanol and tri-metaphosphate. The reduction rate of U(VI) depends on sulfate concentration in water and the abundance of bacteria in the system. This work shows that bacteria capable of U(VI) reduction are ubiquitous in nature, and suggests the possibility of a large application of the enzymatic reduction of U(VI) for in situ clean up of groundwaters contaminated with uranium. (authors)

  2. Rational design of functional and tunable oscillating enzymatic networks

    Science.gov (United States)

    Semenov, Sergey N.; Wong, Albert S. Y.; van der Made, R. Martijn; Postma, Sjoerd G. J.; Groen, Joost; van Roekel, Hendrik W. H.; de Greef, Tom F. A.; Huck, Wilhelm T. S.

    2015-02-01

    Life is sustained by complex systems operating far from equilibrium and consisting of a multitude of enzymatic reaction networks. The operating principles of biology's regulatory networks are known, but the in vitro assembly of out-of-equilibrium enzymatic reaction networks has proved challenging, limiting the development of synthetic systems showing autonomous behaviour. Here, we present a strategy for the rational design of programmable functional reaction networks that exhibit dynamic behaviour. We demonstrate that a network built around autoactivation and delayed negative feedback of the enzyme trypsin is capable of producing sustained oscillating concentrations of active trypsin for over 65 h. Other functions, such as amplification, analog-to-digital conversion and periodic control over equilibrium systems, are obtained by linking multiple network modules in microfluidic flow reactors. The methodology developed here provides a general framework to construct dissipative, tunable and robust (bio)chemical reaction networks.

  3. From Fed-batch to Continuous Enzymatic Biodiesel Production

    DEFF Research Database (Denmark)

    Price, Jason Anthony; Nordblad, Mathias; Woodley, John M.

    2015-01-01

    In this this paper, we use mechanistic modelling to guide the development of acontinuous enzymatic process that is performed as a fed-batch operation. In this workwe use the enzymatic biodiesel process as a case study. A mechanistic model developedin our previous work was used to determine...... measured components (triglycerides, diglycerides, monoglycerides, free fatty acid and fatty acid methyl esters(biodiesel)) much better than using fed-batch data alone given the smaller residuals. We also observe a reduction in the correlation between the parameters.The model was then used to predict that 5...... reactors are required (with a combined residence time of 30 hours) to reach a final biodiesel concentration within 2 % of the95.6 mass % achieved in a fed-batch operation, for 24 hours....

  4. Novel investigation of enzymatic biodiesel reaction by isothermal calorimetry

    DEFF Research Database (Denmark)

    Søtoft, Lene Fjerbaek; Westh, Peter; Christensen, Knud V.

    2010-01-01

    Isothermal calorimetry (ITC) was used to investigate solvent-free enzymatic biodiesel production. The transesterification of rapeseed oil with methanol and ethanol was catalyzed by immobilized lipase Novozym 435 at 40 °C. The aim of the study was to determine reaction enthalpy for the enzymatic...... transesterification and to elucidate the mass transfer and energetic processes taking place. Based on the measured enthalpy and composition change in the system, the heat of reaction at 40 °C for the two systems was determined as −9.8 ± 0.9 kJ/mole biodiesel formed from rapeseed oil and methanol, and −9.3 ± 0.7 k...

  5. Structural Characterization and Enzymatic Modification of Soybean Polysaccharides

    DEFF Research Database (Denmark)

    Pierce, Brian; Wichmann, Jesper

    % galacturonic acid, 8% xylose, 3% rhamnose, and 3% fucose. Currently, the majority of this material is disposed of as waste, increasing production costs. Opportunities exist for the develop-ment of novel functional ingredients from this abundant and underutilized ma-terial; however, efforts in this area......The work in this thesis explores the structure of soybean polysaccharides, and examines approaches for the chemical and enzymatic degradation and solu-bilization of this material. Soybean polysaccharides are produced in large quantities globally as a by-product of various soy production processes...... are currently limited by the material’s insol-ubility. A central hypothesis of this work was that by obtaining a more complete understanding of the structure of this material, chemical and enzymatic ap-proaches could be developed to modify the polysaccharides, creating soluble polysaccharide fractions...

  6. Non-enzymatic palladium recovery on microbial and synthetic surfaces

    DEFF Research Database (Denmark)

    Rotaru, Amelia-Elena; Jiang, Wei; Finster, Kai

    2012-01-01

    in the presence of cells as compared to cell-free controls. We found no difference between native (untreated) and autoclaved cells, and could demonstrate that even a non-enzymatic protein (bovine serum albumin) stimulated Pd(II) reduction as efficiently as bacterial cells. Amine groups readily interact with Pd......(II), and to specifically test their role in surface-assisted Pd(II) reduction by formate, we replaced bacterial cells with polystyrene microparticles functionalized with amine or carboxyl groups. Amine-functionalized microparticles had the same effect on Pd(II) reduction as bacterial cells, and the effect could...... be hampered if the amine groups were blocked by acetylation. The interaction with amine groups was confirmed by infrared spectroscopy on whole cells and amine-functionalized microparticles. In conclusion, bio-supported Pd(II) reduction on microbial surfaces is possibly mediated by a non-enzymatic mechanism...

  7. Enzymatic network for production of ether amines from alcohols

    DEFF Research Database (Denmark)

    Palacio, Cyntia M.; Crismaru, Ciprian G.; Bartsch, Sebastian

    2016-01-01

    We constructed an enzymatic network composed of three different enzymes for the synthesis of valuable ether amines. The enzymatic reactions are interconnected to catalyze the oxidation and subsequent transamination of the substrate and to provide cofactor recycling. This allows production...... of the desired ether amines from the corresponding ether alcohols with inorganic ammonium as the only additional substrate. To examine conversion, individual and overall reaction equilibria were established. Using these data, it was found that the experimentally observed conversions of up to 60% observed...... for reactions containing 10mM alcohol and up to 280mM ammonia corresponded well to predicted conversions. The results indicate that efficient amination can be driven by high concentrations of ammonia and may require improving enzyme robustness for scale-up....

  8. Graphene paper based bioelectrodes for enzymatic biofuel cells

    DEFF Research Database (Denmark)

    Werchmeister, Rebecka Maria Larsen; Shen, Fei; Zhang, Jingdong

    We aim at developing bioelectrodes for enzymatic biofuel cells, where sustainable and renewable enzymes are used for catalyzing the oxidation and reduction of fuel molecules. Here glucose is chosen as fuel molecule and glucose oxidase (GOx) is target enzyme which catalyzes the oxidation of glucose...... of glucose. This indicates that the enzyme has been successfully immobilized and is actively consuming glucose while transferring electrons to the graphene paper-GOx bioanode. Stability and efficiency of the bioelectrodes are under investigation....

  9. Apple phenolics and their contribution to enzymatic browning reactions

    Directory of Open Access Journals (Sweden)

    Wiesław Oleszek

    2014-01-01

    Full Text Available Chlorogenic acid, epicatechin, procyanidin B2 and C1 were isolated from apple skin. These compounds as well as quercetine and phloretine glycosides isolated from apples were studied individually and as mixtures for their participation in the enzymatic browning reactions. The importance of quercetine glycosides and the synergistic effect of phloridzin and phloretine xyloglucoside with chlorogenic acid and flavans in the browning reaction are reported.

  10. Nanosilver: A Catalyst in Enzymatic Hydrolysis of Starch

    Directory of Open Access Journals (Sweden)

    Falkowska Marta

    2014-09-01

    Full Text Available Silver nanoparticles are widely used, because of their antimicrobial properties. In this paper, the rate of starch digestion in the presence of nanocatalyst was compared with the rate of reaction without nanosilver. The rate of enzymatic degradation of starch was found to be increased in the presence of silver nanoparticles. It is considered that α-amylase was immobilized onto the surface of nanoparticles.

  11. Recent Trends in Quantum Chemical Modeling of Enzymatic Reactions.

    Science.gov (United States)

    Himo, Fahmi

    2017-05-24

    The quantum chemical cluster approach is a powerful method for investigating enzymatic reactions. Over the past two decades, a large number of highly diverse systems have been studied and a great wealth of mechanistic insight has been developed using this technique. This Perspective reviews the current status of the methodology. The latest technical developments are highlighted, and challenges are discussed. Some recent applications are presented to illustrate the capabilities and progress of this approach, and likely future directions are outlined.

  12. Novel flavonolignan hybrid antioxidants: From enzymatic preparation to molecular rationalization

    Czech Academy of Sciences Publication Activity Database

    Vavříková, Eva; Křen, Vladimír; Ježová-Kalachová, Lubica; Biler, M.; Chantemargue, B.; Pyszková, M.; Riva, S.; Kuzma, Marek; Valentová, Kateřina; Ulrichová, J.; Vrba, J.; Trouillas, P.; Vacek, J.

    2017-01-01

    Roč. 127, FEB 15 (2017), s. 263-274 ISSN 0223-5234 R&D Projects: GA MŠk(CZ) LD14096; GA MŠk(CZ) LD15084; GA ČR(CZ) GA15-03037S Institutional support: RVO:61388971 Keywords : Silybin * Vitamin C * Enzymatic coupling Subject RIV: CE - Biochemistry OBOR OECD: Biochemistry and molecular biology Impact factor: 4.519, year: 2016

  13. Global Warming Potential Of A Waste Refinery Using Enzymatic Treatment

    DEFF Research Database (Denmark)

    Tonini, Davide; Astrup, Thomas

    2010-01-01

    and fossil resources. This is especially important with respect to the residual waste (i.e. the remains after source-separation and separate collection) which is typically incinerated or landfilled. In this paper the energy and Global Warming performance of a pilot-scale waste refinery for the enzymatic...... plants and utilization of the liquid fraction for biogas production turned out to be the best options with respect to energy and Global Warming performance....

  14. ASPECTS CONCERNING THE ENZYMATIC ACTIVITY IN SEVERAL THERMOACTINOMYCETE STRAINS

    Directory of Open Access Journals (Sweden)

    Simona Dunca

    2003-08-01

    Full Text Available In the thermoactinomycete strains subjected to examination the values of their recorded enzymatic activities (i.e. α-amy lase, protease, exo-β-1,4 – glucanase, endo -β-1,4 – glucanase and β-glucosidase were lower in the stationary cultures as compared to the stirred ones. The strain Thermomonospora fusca BB255 was found to be highly cellulase- producing and at the same time able to synthesize α-amy lases and proteases.

  15. Enzymatic determination of rare earth elements using pyrophosphatases

    International Nuclear Information System (INIS)

    Shekhovtsova, T.N.; Pirogova, S.V.; Fedorova, O.M.; Dolmanova, I.F.; Bajkov, A.A.

    1993-01-01

    A highly sensitive(determination limit 8x10 -6 -4x10 -4 μ g/m) and selective enzymatic method for determination of rare earth elements has been developed. The method is based on inhibition action of rare earths on the catalytic activity of pyrophosphates isolated from bakery geast and E.Coli. The mechanism of the rare earth element action, corresponding to competitive inhibition, has been established

  16. Enzymatic determination of rare earth elements by use of pyrophosphotases

    International Nuclear Information System (INIS)

    Shekhovtseva, T.N.; Pirogova, S.V.; Fedorova, O.M.; Dolmanova, I.F.; Bajkov, A.A.

    1993-01-01

    A highly sensitive (determination limit 8 x 10 -6 - 4 x 10 -4 μg/ml) and selective enzymatic method for determination of rare earth elements has been developed. The method is based on inhibition action of rare earths on the catalytic activity of pyrophosphates isolated from bakery geast and E. Coli. The mechanism of the rare earth element action, corresponding to competitive inhibition, has been established

  17. Enzymatic preparation and characterization of soybean lecithin-based emulsifiers

    OpenAIRE

    R. C. Reddy Jala; B. Chen; H. Li; Y. Zhang; L-Z Cheong; T. Yang; X. Xu

    2016-01-01

    Simple enzymatic methods were developed for the synthesis of lysolecithin, glycerolyzed lecithin and hydrolyzed lecithin. The products were characterized in terms of their acetone insoluble matter, hexane insoluble matter, moisture, phospholipid distribution and fatty acid composition. The HLB value ranges of different products with different acid values were detected. The efficiency of optimally hydrolyzed lecithin was examined at high calcium ion, low pH, and aqueous solutions and compared ...

  18. Bioethanol production: Pretreatment and enzymatic hydrolysis of softwood

    Energy Technology Data Exchange (ETDEWEB)

    Tengborg, Charlotte

    2000-05-01

    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.

  19. Inhibition of peptide aggregation by means of enzymatic phosphorylation

    Directory of Open Access Journals (Sweden)

    Kristin Folmert

    2016-11-01

    Full Text Available As is the case in numerous natural processes, enzymatic phosphorylation can be used in the laboratory to influence the conformational populations of proteins. In nature, this information is used for signal transduction or energy transfer, but has also been shown to play an important role in many diseases like tauopathies or diabetes. With the goal of determining the effect of phosphorylation on amyloid fibril formation, we designed a model peptide which combines structural characteristics of α-helical coiled-coils and β-sheets in one sequence. This peptide undergoes a conformational transition from soluble structures into insoluble amyloid fibrils over time and under physiological conditions and contains a recognition motif for PKA (cAMP-dependent protein kinase that enables enzymatic phosphorylation. We have analyzed the pathway of amyloid formation and the influence of enzymatic phosphorylation on the different states along the conformational transition from random-coil to β-sheet-rich oligomers to protofilaments and on to insoluble amyloid fibrils, and we found a remarkable directing effect from β-sheet-rich structures to unfolded structures in the initial growth phase, in which small oligomers and protofilaments prevail if the peptide is phosphorylated.

  20. Metal nanostructures for non-enzymatic glucose sensing

    International Nuclear Information System (INIS)

    Tee, Si Yin; Teng, Choon Peng; Ye, Enyi

    2017-01-01

    This review covers the recent development of metal nanostructures in electrochemical non-enzymatic glucose sensing. It highlights a variety of nanostructured materials including noble metals, other transition metals, bimetallic systems, and their hybrid with carbon-based nanomaterials. Particularly, attention is devoted to numerous approaches that have been implemented for improving the sensors performance by tailoring size, shape, composition, effective surface area, adsorption capability and electron-transfer properties. The correlation of the metal nanostructures to the glucose sensing performance is addressed with respect to the linear concentration range, sensitivity and detection limit. In overall, this review provides important clues from the recent scientific achievements of glucose sensor nanomaterials which will be essentially useful in designing better and more effective electrocatalysts for future electrochemical sensing industry. - Highlights: • Overview of recent development of metal nanostructures in electrochemical non-enzymatic glucose sensing. • Special attention is focussed on noble metals, other transition metals, bimetallic systems, and their hybrid with carbon-based nanomaterials. • Merits and limitations of various metal nanostructures in electrochemical non-enzymatic glucose sensing. • Strategies to improve the glucose sensing performance of metal nanostructures as electrocatalysts.

  1. Microstructural study of pre-treated and enzymatic hydrolyzed bamboo

    Directory of Open Access Journals (Sweden)

    Funsho O. KOLAWOLE

    2016-07-01

    Full Text Available Bamboo was used as biomass feedstock which was pre-treated using dilute acid hydrolysis followed by enzymatic hydrolysis. The bamboo was mechanical ground to particle sizes 212–500µm, followed by pre-treatment with dilute sulfuric acid at a concentration of 0.5 and 1.0 (%v/v at temperatures of 25, 110, 120, 150 and 200°C with time intervals of 2 and 4 hours. Pre-hydrolyzate was later analyzed for reducing sugar using UV-Vis spectrophotometry. Under the above conditions, a maximum glucose yield of 153.1 mg/g was obtained at 200°C and acid concentrations of 1% for 4 hours. Water insoluble solids obtained were subsequently hydrolyzed with Celluclast (Trichoderma reesi and β-glucosidase (Novozyme 188 for 72 hours. Optical Microscope and ESEM images of bamboo samples were obtained at various stages of pre-treatment and enzymatic hydrolysis. Result reveals a breakdown in the ligno-cellulosic structure of the bamboo during exposure to dilute acid and enzymatic hydrolysis.

  2. Enzymatic determination of cadmium, zinc, and lead in plant materials

    International Nuclear Information System (INIS)

    Muginova, S.V.; Veselova, I.A.; Parova, L.M.; Shekhovtseva, T.N.

    2008-01-01

    Prospects are outlined for using the following enzymes (native and immobilized on polyurethane foam) in the rapid and highly sensitive determination of cadmium, zinc, and lead ions in plant materials (wild grass, fresh pea, and grape): horseradish peroxidase and alkaline phosphatases isolated from chicken intestine and Greenland seal small intestine. The analytical ranges of the above metals are 1x10 -3 -25; 7x10 -3 -250, and 3x10 -2 -67 mg/kg dry matter, respectively. The enzymatic determination procedures developed are based on the inhibiting effect of metal ions on the catalytic activity of peroxidase in the oxidation of o-dianisidine with hydrogen peroxide and alkaline phosphatases in the hydrolysis of p-nitrophenyl phosphate. The rates of enzymatic reactions were monitored spectrophotometrically or visually. In the analysis of plant extracts, their high acidity was diminished by choosing optimum dilution factors and pH values for test samples and the nature and concentration of a buffer solution. The interference of iron(III) was removed by introducing a 0.1 M tartaric acid solution into the indicator reaction. The accuracy of the results of the enzymatic determination of cadmium, zinc, and lead in plant materials was supported by atomic absorption spectrometry and anodic stripping voltammetry [ru

  3. Comparison of the suitability of alkaline or enzymatic sample pre-treatment for characterization of silver nanoparticles in human tissue by single particle ICP-MS

    DEFF Research Database (Denmark)

    Vidmar, Janja; Buerki-Thurnherr, Tina; Löschner, Katrin

    2018-01-01

    and their size are required for studying NP accumulation in placental tissue. In the present study, we applied and compared two sample preparation techniques, alkaline and enzymatic treatment, followed by single particle ICP-MS (spICP-MS) analysis, for characterizing AgNPs spiked to human placental tissue. Both...... sample preparation approaches are currently used for AgNPs in biological tissues but have not been directly compared yet. We showed that the method using enzymatic tissue treatment followed by spICP-MS is efficient for determination of mass and number concentration and size distribution of AgNPs in human...... placental tissues. Properties of the AgNPs were preserved during enzymatic digestion and comparable with the primary particles. The matrix effect on the determination of Ag sensitivity and transport efficiency in spICP-MS analysis was systematically evaluated as well. The method was applied to human...

  4. Physical and sensory characteristics of pork sausages from enzymatically modified blends of lard and rapeseed oil during storage

    DEFF Research Database (Denmark)

    Cheong, L.Z.; Zhang, H.; Nersting, L.

    2010-01-01

    Physical and sensory characteristic of pork sausages produced from enzymatic interesterified blends of lard and rapeseed oil during storage were evaluated. All three enzymatic interesterified blends (IE90, IE70 and IE50) had ratios of unsaturated to saturated fatty acids within the range of 1.......47-2.84 which is favourable for cardiovascular disease risk reduction. Blends of IE90 and IE70 were found to have suitable solid fat content, melting and crystallization profile suitable for sausages production. Sausages were produced from blends of IE90 and IE70 with different muscle types (musculus...... longissimus dorsi and musculus sternomandibularis) and processing conditions such as cooling rates and final processing temperature. Cooling rate was found to have no significant (P>0.05) effect on hardness of the sausages throughout storage. Both musculus longissimus dorsi and high final processing...

  5. Removal of antibiotics in wastewater by enzymatic treatment with fungal laccase - Degradation of compounds does not always eliminate toxicity.

    Science.gov (United States)

    Becker, Dennis; Varela Della Giustina, Saulo; Rodriguez-Mozaz, Sara; Schoevaart, Rob; Barceló, Damià; de Cazes, Matthias; Belleville, Marie-Pierre; Sanchez-Marcano, José; de Gunzburg, Jean; Couillerot, Olivier; Völker, Johannes; Oehlmann, Jörg; Wagner, Martin

    2016-11-01

    In this study, the performance of immobilised laccase (Trametes versicolor) was investigated in combination with the mediator syringaldehyde (SYR) in removing a mixture of 38 antibiotics in an enzymatic membrane reactor (EMR). Antibiotics were spiked in osmosed water at concentrations of 10μg·L(-1) each. Laccase without mediator did not reduce the load of antibiotics significantly. The addition of SYR enhanced the removal: out of the 38 antibiotics, 32 were degraded by >50% after 24h. In addition to chemical analysis, the samples' toxicity was evaluated in two bioassays (a growth inhibition assay and the Microtox assay). Here, the addition of SYR resulted in a time-dependent increase of toxicity in both bioassays. In cooperation with SYR, laccase effectively removes a broad range of antibiotics. However, this enhanced degradation induces unspecific toxicity. If this issue is resolved, enzymatic treatment may be a valuable addition to existing water treatment technologies. Copyright © 2016 Elsevier Ltd. All rights reserved.

  6. Characterization of Enzymatic profiles of Aedes aegypti strains from the State of Rio Grande do Norte, Brazil

    Directory of Open Access Journals (Sweden)

    Renan Flávio de França Nunes

    2016-01-01

    Full Text Available Abstract This study was conducted in four strains of Aedes aegypti mosquitoes to evaluate the enzymatic activity profiles in the city of Mossoró, Rio Grande do Norte, and correlate them with biochemical mechanisms of resistance to insecticides. Mosquitos were used to quantify the following detoxification enzymes: Mixed-Function Oxidase (MFO, PNPA-esterase (PNPA-EST, and Acetylcholinesterase (AChE. The profiles were compared statistically with profiles from the Rockefeller strain, through the Kruskal-Wallis test and Dunn's multiple comparisons (p 15% and 50%. The statistical analysis revealed significant differences in the MFO and AChE profiles, which are fundamental in the determination of profiles of resistance to insecticides. Three populations were classified as “Substantially changed” for MFO. The altered enzymatic activity showed that the changes could have an important role in exposing resistance to insecticides.

  7. Comparison of non-volatile umami components in chicken soup and chicken enzymatic hydrolysate.

    Science.gov (United States)

    Kong, Yan; Yang, Xiao; Ding, Qi; Zhang, Yu-Yu; Sun, Bao-Guo; Chen, Hai-Tao; Sun, Ying

    2017-12-01

    Umami taste is an important part to the taste of chicken. To isolate and identify non-volatile umami compounds, fractions from chicken soup and hydrolysate were prepared and analyzed. Amino acids were analyzed by amino acid analyzer. Organic acids and nucleotides were determined by ultra-performance liquid chromatography. Separation procedures utilizing ultrafiltration, Sephadex G-15 and reversed-phase high-performance liquid chromatography were used to isolate umami taste peptides. Combined with sensory evaluation and LC-Q-TOF-MS, the amino acid sequences of 12 oligopeptides were determined. The amount of taste compounds was higher in chicken enzymatic hydrolysate than that of chicken soup. Eight oligopeptides from chicken enzymatic hydrolysate were identified, including Ala-Asp, Ala-Met, His-Ser, Val-Glu, Ala-Glu, Asp-Ala-Gly, Glu-Asp and Ala-Glu-Ala. Four oligopeptides from chicken soup were identified, including Val-Thr, Ala-His, Ala-Phe and Thr-Glu. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Alkali-aided enzymatic viscosity reduction of sugar beet mash for novel bioethanol production process

    International Nuclear Information System (INIS)

    Srichuwong, Sathaporn; Arakane, Mitsuhiro; Fujiwara, Maki; Zhang, Zilian; Takahashi, Hiroyuki; Tokuyasu, Ken

    2010-01-01

    Ethanol fermentation of fresh sugar beet mash (SBM) could give a benefit on reducing energy input for sugar diffusion, juice separation, and water evaporation as used in conventional practices, thus offering promise as a low energy process. Actions of cell-wall degrading enzymes provide a mash with low viscosity, which can be easily fermented to ethanol. However, a several-fold higher enzyme loading was required for viscosity reduction of SBM compared with that of potato mash. In this study, the use of dilute alkali treatment (0.025-0.15 N NaOH, 25 o C, 1 h) in enhancing enzymatic viscosity reduction of SBM was evaluated. The results showed that higher NaOH concentration enhanced demethylation and deacetylation of SBM, resulting in greater performances of the enzymes on reducing viscosity. Efficient enzymatic viscosity reduction of SBM was observed with the 0.1 N NaOH treatment. On the other hand, untreated SBM was highly resistant to viscosity reduction, even though a 20-fold more enzyme loading was used. The resulting mash containing 12-13% (w/v) sucrose yielded 7-8% (v/v) ethanol after 24 h of fermentation (90% efficiency). Accordingly, alkali treatment can be applied for facilitating the use of fresh sugar beet for ethanol production.

  9. Effect of depth beating on the fiber properties and enzymatic saccharification efficiency of softwood kraft pulp.

    Science.gov (United States)

    Gao, Wenhua; Xiang, Zhouyang; Chen, Kefu; Yang, Rendang; Yang, Fei

    2015-01-01

    Commercial bleached softwood kraft pulp was mechanically fibrillated by a PFI-mill with beating revolution from 5000 to 30,000 r. The extent of fibrillating on the pulp was evaluated by beating degree, fiber morphological properties (fiber length, width, coarseness and curls index), water retention value (WRV) and physical properties of paper made from the pulp. Depth beating process significantly affected the pulp fibrillations as showed by the decreased fiber length and width as well as the SEM analysis, but the effects were limited after beating revolution of 15,000. Depth beating process also improved the total internal pore and inter-fibril surface areas as shown by the increased WRV values. Substrate enzymatic digestibility (SED) of beaten pulp at 5000 revolutions could reach 95% at cellulase loading of 15 FPU/g of glucan. After the enzymatic hydrolysis, the size of the pulp residues was reduced to micro-scale, and a relative uniform size distribution of the residues appeared at 10,000 r beating revolution. Copyright © 2015 Elsevier Ltd. All rights reserved.

  10. Relationship between Porcine Sperm Motility and Sperm Enzymatic Activity using Paper-based Devices

    Science.gov (United States)

    Matsuura, Koji; Huang, Han-Wei; Chen, Ming-Cheng; Chen, Yu; Cheng, Chao-Min

    2017-04-01

    Mammalian sperm motility has traditionally been analyzed to determine fertility using computer-assisted semen analysis (CASA) systems. To develop low-cost and robust male fertility diagnostics, we created a paper-based MTT assay and used it to estimate motile sperm concentration. When porcine sperm motility was inhibited using sperm enzyme inhibitors for sperm enzymes related to mitochondrial activity and glycolysis, we simultaneously recorded sperm motility and enzymatic reactivity using a portable motility analysis system (iSperm) and a paper-based MTT assay, respectively. When using our paper-based MTT-assay, we calculated the area mean value signal intensity (AMV) to evaluate enzymatic reactivity. Both sperm motility and AMV decreased following treatment with iodoacetamide (IODO) and 3-bromopyruvic acid (3BP), both of which are inhibitors of glycolytic enzymes including glyceraldehyde-3-phosphate dehydrogenase (GAPDH). We found a correlation between recorded motility using iSperm and AMV from our paper-based assay (P Based on this inhibitor study, sperm motility can be estimated using our paper-based MTT-assay.

  11. Sporothrix schenckii COMPLEX: SUSCEPTIBILITIES TO COMBINED ANTIFUNGAL AGENTS AND CHARACTERIZATION OF ENZYMATIC PROFILES

    Directory of Open Access Journals (Sweden)

    Daniele Carvalho OLIVEIRA

    2015-08-01

    Full Text Available SUMMARY Sporothrix schenckiiwas reclassified as a complex encompassing six cryptic species, which calls for the reassessment of clinical and epidemiological data of these new species. We evaluated the susceptibility of Sporothrix albicans (n = 1 , S. brasiliensis (n = 6 , S. globosa (n = 1, S. mexicana(n = 1 and S. schenckii(n = 36 to terbinafine (TRB alone and in combination with itraconazole (ITZ, ketoconazole (KTZ, and voriconazole (VRZ by a checkerboard microdilution method and determined the enzymatic profile of these species with the API-ZYM kit. Most interactions were additive (27.5%, 32.5% and 5% or indifferent (70%, 50% and 52.5% for TRB+KTZ, TRB+ITZ and TRB+VRZ, respectively. Antagonisms were observed in 42.5% of isolates for the TRB+VRZ combination. Based on enzymatic profiling, the Sporothrix schenckii strains were categorized into 14 biotypes. Leucine arylamidase (LA activity was observed only for S. albicans and S. mexicana. The species S. globosaand S. mexicanawere the only species without β-glucosidase (GS activity. Our results may contribute to a better understanding of virulence and resistance among species of the genus Sporothrixin further studies.

  12. Sporothrix schenckii COMPLEX:SUSCEPTIBILITIES TO COMBINED ANTIFUNGAL AGENTS AND CHARACTERIZATION OF ENZYMATIC PROFILES.

    Science.gov (United States)

    Oliveira, Daniele Carvalho; de Loreto, Érico Silva; Mario, Débora Alves Nunes; Lopes, Paulo G Markus; Neves, Louise Vignolles; da Rocha, Marta Pires; Santurio, Janio Morais; Alves, Sydney Hartz

    2015-01-01

    Sporothrix schenckii was reclassified as a complex encompassing six cryptic species, which calls for the reassessment of clinical and epidemiological data of these new species. We evaluated the susceptibility of Sporothrix albicans(n = 1) , S. brasiliensis(n = 6) , S. globosa(n = 1), S. mexicana(n = 1) and S. schenckii(n = 36) to terbinafine (TRB) alone and in combination with itraconazole (ITZ), ketoconazole (KTZ), and voriconazole (VRZ) by a checkerboard microdilution method and determined the enzymatic profile of these species with the API-ZYM kit. Most interactions were additive (27.5%, 32.5% and 5%) or indifferent (70%, 50% and 52.5%) for TRB+KTZ, TRB+ITZ and TRB+VRZ, respectively. Antagonisms were observed in 42.5% of isolates for the TRB+VRZ combination. Based on enzymatic profiling, the Sporothrix schenckii strains were categorized into 14 biotypes. Leucine arylamidase (LA) activity was observed only for S. albicans and S. mexicana. The species S. globosa and S. Mexicana were the only species without β-glucosidase (GS) activity. Our results may contribute to a better understanding of virulence and resistance among species of the genus Sporothrix in further studies.

  13. Colorimetric determination of nitrate plus nitrite in water by enzymatic reduction, automated discrete analyzer methods

    Science.gov (United States)

    Patton, Charles J.; Kryskalla, Jennifer R.

    2011-01-01

    This report documents work at the U.S. Geological Survey (USGS) National Water Quality Laboratory (NWQL) to validate enzymatic reduction, colorimetric determinative methods for nitrate + nitrite in filtered water by automated discrete analysis. In these standard- and low-level methods (USGS I-2547-11 and I-2548-11), nitrate is reduced to nitrite with nontoxic, soluble nitrate reductase rather than toxic, granular, copperized cadmium used in the longstanding USGS automated continuous-flow analyzer methods I-2545-90 (NWQL laboratory code 1975) and I-2546-91 (NWQL laboratory code 1979). Colorimetric reagents used to determine resulting nitrite in aforementioned enzymatic- and cadmium-reduction methods are identical. The enzyme used in these discrete analyzer methods, designated AtNaR2 by its manufacturer, is produced by recombinant expression of the nitrate reductase gene from wall cress (Arabidopsis thaliana) in the yeast Pichia pastoris. Unlike other commercially available nitrate reductases we evaluated, AtNaR2 maintains high activity at 37°C and is not inhibited by high-phenolic-content humic acids at reaction temperatures in the range of 20°C to 37°C. These previously unrecognized AtNaR2 characteristics are essential for successful performance of discrete analyzer nitrate + nitrite assays (henceforth, DA-AtNaR2) described here.

  14. Role of conformational dynamics in kinetics of an enzymatic cycle in a nonequilibrium steady state

    Science.gov (United States)

    Min, Wei; Xie, X. Sunney; Bagchi, Biman

    2009-08-01

    Enzyme is a dynamic entity with diverse time scales, ranging from picoseconds to seconds or even longer. Here we develop a rate theory for enzyme catalysis that includes conformational dynamics as cycling on a two-dimensional (2D) reaction free energy surface involving an intrinsic reaction coordinate (X) and an enzyme conformational coordinate (Q). The validity of Michaelis-Menten (MM) equation, i.e., substrate concentration dependence of enzymatic velocity, is examined under a nonequilibrium steady state. Under certain conditions, the classic MM equation holds but with generalized microscopic interpretations of kinetic parameters. However, under other conditions, our rate theory predicts either positive (sigmoidal-like) or negative (biphasic-like) kinetic cooperativity due to the modified effective 2D reaction pathway on X-Q surface, which can explain non-MM dependence previously observed on many monomeric enzymes that involve slow or hysteretic conformational transitions. Furthermore, we find that a slow conformational relaxation during product release could retain the enzyme in a favorable configuration, such that enzymatic turnover is dynamically accelerated at high substrate concentrations. The effect of such conformation retainment in a nonequilibrium steady state is evaluated.

  15. Particle size distribution of rice flour affecting the starch enzymatic hydrolysis and hydration properties.

    Science.gov (United States)

    de la Hera, Esther; Gomez, Manuel; Rosell, Cristina M

    2013-10-15

    Rice flour is becoming very attractive as raw material, but there is lack of information about the influence of particle size on its functional properties and starch digestibility. This study evaluates the degree of dependence of the rice flour functional properties, mainly derived from starch behavior, with the particle size distribution. Hydration properties of flours and gels and starch enzymatic hydrolysis of individual fractions were assessed. Particle size heterogeneity on rice flour significantly affected functional properties and starch features, at room temperature and also after gelatinization; and the extent of that effect was grain type dependent. Particle size heterogeneity on rice flour induces different pattern in starch enzymatic hydrolysis, with the long grain having slower hydrolysis as indicated the rate constant (k). No correlation between starch digestibility and hydration properties or the protein content was observed. It seems that in intact granules interactions with other grain components must be taken into account. Overall, particle size fractionation of rice flour might be advisable for selecting specific physico-chemical properties. Copyright © 2013. Published by Elsevier Ltd.

  16. Highly selective apo-arginase based method for sensitive enzymatic assay of manganese (II) and cobalt (II) ions

    Science.gov (United States)

    Stasyuk, Nataliya; Gayda, Galina; Zakalskiy, Andriy; Zakalska, Oksana; Errachid, Abdelhamid; Gonchar, Mykhailo

    2018-03-01

    A novel enzymatic method of manganese (II) and cobalt (II) ions assay, based on using apo-enzyme of Mn2 +-dependent recombinant arginase I (arginase) and 2,3-butanedione monoxime (DMO) as a chemical reagent is proposed. The principle of the method is the evaluation of the activity of L-arginine-hydrolyzing of arginase holoenzyme after the specific binding of Mn2 + or Co2 + with apo-arginase. Urea, which is the product of enzymatic hydrolysis of L-arginine (Arg), reacts with DMO and the resulted compound is detected by both fluorometry and visual spectrophotometry. Thus, the content of metal ions in the tested samples can be determined by measuring the level of urea generated after enzymatic hydrolysis of Arg by reconstructed arginase holoenzyme in the presence of tested metal ions. The linearity range of the fluorometric apo-arginase-DMO method in the case of Mn2 + assay is from 4 pM to 1.10 nM with a limit of detection of 1 pM Mn2 +, whereas the linearity range of the present method in the case of Co2 + assay is from 8 pM to 45 nM with a limit of detection of 2.5 pM Co2 +. The proposed method being highly sensitive, selective, valid and low-cost, may be useful to monitor Mn2 + and Co2 + content in clinical laboratories, food industry and environmental control service.

  17. Effect of enzymatic pretreatment on the physical quality of plantain (Musa ssp., group AAB) employing airflow reversal drying.

    Science.gov (United States)

    Rodríguez-Miranda, J; Martínez-Sánchez, C E; Hernández-Santos, B; Juárez-Barrientos, J M; Ventura-Báez, E G; Herman-Lara, E

    2018-01-01

    This work aimed to evaluate the effect of enzymatic pretreatment on the color and texture of plantain ( Musa ssp., group AAB) dried by airflow reversal drying. Plantain slices 1.0 cm thick were used. Pretreatment with two commercial enzymes, Pectinex Ultra SPL ( Aspergillus aculeatus ) and Pectinex 3XL ( Aspergillus niger ), was performed. Drying kinetics were determined with and without pretreatment at temperatures of 50, 65 and 80 °C using a fixed bed convective dryer. An air speed of 6 m/s, a bed height of 5 cm and either unidirectional flow or airflow reversal (every 15 min) were used for drying. Color and texture were analyzed, and consumer acceptance of the results of the best treatments was determined. Pretreatment with the enzyme A. niger and airflow reversal gave the best drying kinetics and showed the greatest reduction in drying time (59.0%) at 80 °C. The best hardness results were found at 80 °C with A. niger enzymatic pretreatment with both types of air flow. Brightness and hue angle showed that samples pretreated with enzymes and dried at 65 °C had a lighter yellow color compared to non-pretreated samples. Plantain samples enzymatically pretreated and dried at 65 and 80 °C were the most accepted by consumers. This kind of enzymatic pretreatment on plantain could allow the conservation of some physical properties and reduction of drying times relative to the current methodology.

  18. Pregnancy Exercise Increase Enzymatic Antioxidant In Pregnant Women

    Directory of Open Access Journals (Sweden)

    Wagey Freddy Wagey

    2012-01-01

    Full Text Available Objectives: Pregnancy is a vulnerable condition to all kinds of "stress", resulting in changes of physiological and metabolic functions. This research aims to determine effect of exercise during pregnancy in increasing enzymatic antioxidant marked by increase of superoxide dismutase (SOD, gluthation peroxidase (GSHPx, and catalase (CAT levels. Methods: Randomized pre and posttest control group design was employed in this study. A number of 66 pregnant women were recruited in this study and grouped into two groups, i.e 30 of them as control group and the rest as treatment group. Pregnancy exercise was performed to all 36 pregnant women from 20 weeks gestation on treatment group. The exercise was performed in the morning for about 30 minutes, twice a weeks. On the other hand, daily activities was sugested for control group. Student’s t-test was then applied to determine the mean different of treatment and control group with 5 % of significant value. Results: This study reveals that there were significantly higher increase of (superoxide dismutase (SOD, gluthation peroxidase (GSHPx, and catalse (CAT levels of treatment group compare to control group. These enzymatic antioxidant increase among these two group were around 1.36 mg/gHb for SOD; 1.14 IU/gHb for GSHPx; and 0.97 IU/gHb for CAT, (p < 0.05.  Clinical outcomes, such as strengten of pelvic muscle and quality of life of treatment group were significantly better compared to control group (p < 0.05. Conclusions: This means that exercise during pregnancy ages of 20 weeks increase enzymatic antioxidant levels SOD, GSHPx, and CAT higher compare to control group without exercise.  

  19. Pregnancy Exercise Increase Enzymatic Antioxidant In Pregnant Women

    Directory of Open Access Journals (Sweden)

    Wagey Freddy Wagey

    2012-01-01

    Full Text Available Objectives: Pregnancy is a vulnerable condition to all kinds of "stress", resulting in changes of physiological and metabolic functions. This research aims to determine effect of exercise during pregnancy in increasing enzymatic antioxidant marked by increase of superoxide dismutase (SOD, gluthation peroxidase (GSHPx, and catalase (CAT levels. Methods: Randomized pre and posttest control group design was employed in this study. A number of 66 pregnant women were recruited in this study and grouped into two groups, i.e 30 of them as control group and the rest as treatment group. Pregnancy exercise was performed to all 36 pregnant women from 20 weeks gestation on treatment group. The exercise was performed in the morning for about 30 minutes, twice a weeks. On the other hand, daily activities was sugested for control group. Student’s t-test was then applied to determine the mean different of treatment and control group with 5 % of significant value. Results: This study reveals that there were significantly higher increase of (superoxide dismutase (SOD, gluthation peroxidase (GSHPx, and catalse (CAT levels of treatment group compare to control group. These enzymatic antioxidant increase among these two group were around 1.36 mg/gHb for SOD; 1.14 IU/gHb for GSHPx; and 0.97 IU/gHb for CAT, (p < 0.05. Clinical outcomes, such as strengten of pelvic muscle and quality of life of treatment group were significantly better compared to control group (p < 0.05. Conclusions: This means that exercise during pregnancy ages of 20 weeks increase enzymatic antioxidant levels SOD, GSHPx, and CAT higher compare to control group without exercise.

  20. Sugar ester surfactants: enzymatic synthesis and applications in food industry.

    Science.gov (United States)

    Neta, Nair S; Teixeira, José A; Rodrigues, Lígia R

    2015-01-01

    Sugar esters are non-ionic surfactants that can be synthesized in a single enzymatic reaction step using lipases. The stability and efficiency of lipases under unusual conditions and using non-conventional media can be significantly improved through immobilization and protein engineering. Also, the development of de novo enzymes has seen a significant increase lately under the scope of the new field of synthetic biology. Depending on the esterification degree and the nature of fatty acid and/or sugar, a range of sugar esters can be synthesized. Due to their surface activity and emulsifying capacity, sugar esters are promising for applications in food industry.

  1. Multicompartment Artificial Organelles Conducting Enzymatic Cascade Reactions inside Cells

    DEFF Research Database (Denmark)

    Gallardo, Maria Godoy; Labay, Cédric Pierre; Trikalitis, Vasileios

    2017-01-01

    Cell organelles are subcellular structures entrapping a set of enzymes to achieve a specific functionality. The incorporation of artificial organelles into cells is a novel medical paradigm which might contribute to the treatment of various cell disorders by replacing malfunctioning organelles....... In particular, artificial organelles are expected to be a powerful solution in the context of enzyme replacement therapy since enzymatic malfunction is the primary cause of organelle dysfunction. Although several attempts have been made to encapsulate enzymes within a carrier vehicle, only few intracellularly...

  2. Sequential enzymatic epoxidation involved in polyether lasalocid biosynthesis.

    Science.gov (United States)

    Minami, Atsushi; Shimaya, Mayu; Suzuki, Gaku; Migita, Akira; Shinde, Sandip S; Sato, Kyohei; Watanabe, Kenji; Tamura, Tomohiro; Oguri, Hiroki; Oikawa, Hideaki

    2012-05-02

    Enantioselective epoxidation followed by regioselective epoxide opening reaction are the key processes in construction of the polyether skeleton. Recent genetic analysis of ionophore polyether biosynthetic gene clusters suggested that flavin-containing monooxygenases (FMOs) could be involved in the oxidation steps. In vivo and in vitro analyses of Lsd18, an FMO involved in the biosynthesis of polyether lasalocid, using simple olefin or truncated diene of a putative substrate as substrate mimics demonstrated that enantioselective epoxidation affords natural type mono- or bis-epoxide in a stepwise manner. These findings allow us to figure out enzymatic polyether construction in lasalocid biosynthesis. © 2012 American Chemical Society

  3. Radiation degration and the subsequent enzymatic hydrolysis of waste papers

    International Nuclear Information System (INIS)

    Kamakura, M.; Kaetsu, I.

    1982-01-01

    In recent years, many methods have been proposed for the hydrolysis of waste cellulose to utilize it as a new source of alcohol. Because it is difficult to hydrolyze waste cellulosic materials effectivley with an enzyme, the effects of preirradiating waste papers on subsequent enzymatic hydrolysis was studied. Preirradiation (x rays from 60 Co) accelerated the hydrolysis rate of newspaper by cellulase and the reducing-sugar yield increased with increasing irradiation dose. It is thought that preirradiation probably contributes to loosening and releasing the compactly entangled structure of cellulose and lignin in the materials by radiation degradation

  4. Measure of enzymatic activity coincident with 2450 MHz microwave exposure

    Energy Technology Data Exchange (ETDEWEB)

    Ward, T R; Allis, J W; Elder, J A

    1975-09-01

    Enzyme preparations were exposed to microwave radiation at 2450 MHz and enzymatic activity was simultaneously monitored spectrophotometrically with a crossed-beam exposure detection system. Enzymes studied were glucose 6-phosphate dehydrogenase from human red blood cells and yeast, adenylate kinase from rat liver mitochondria and rabbit muscle, and rat liver microsomal NADPH cytochrome c reductase. No difference was found between the specific activity at 25/sup 0/C of unirradiated controls and enzyme preparations irradiated at an absorbed dose rate of 42 W/kg.

  5. Enzymatic preparation and characterization of soybean lecithin-based emulsifiers

    Directory of Open Access Journals (Sweden)

    R. C. Reddy Jala

    2016-12-01

    Full Text Available Simple enzymatic methods were developed for the synthesis of lysolecithin, glycerolyzed lecithin and hydrolyzed lecithin. The products were characterized in terms of their acetone insoluble matter, hexane insoluble matter, moisture, phospholipid distribution and fatty acid composition. The HLB value ranges of different products with different acid values were detected. The efficiency of optimally hydrolyzed lecithin was examined at high calcium ion, low pH, and aqueous solutions and compared with commercially available standard lecithin-based emulsifiers. Overall, lysolecithin powder was proven to be the best emulsifier even at strong and medium acidic conditions.

  6. Enzymatic preparation and characterization of soybean lecithin-based emulsifiers

    International Nuclear Information System (INIS)

    Reddy Jala, R.C.; Chen, B.; Li, H.; Zhang, Y.; Cheong, L.Z.; Yang, T.; Xu, X.

    2016-01-01

    Simple enzymatic methods were developed for the synthesis of lysolecithin, glycerolyzed lecithin and hydrolyzed lecithin. The products were characterized in terms of their acetone insoluble matter, hexane insoluble matter, moisture, phospholipid distribution and fatty acid composition. The HLB value ranges of different products with different acid values were detected. The efficiency of optimally hydrolyzed lecithin was examined at high calcium ion, low pH, and aqueous solutions and compared with commercially available standard lecithin-based emulsifiers. Overall, lysolecithin powder was proven to be the best emulsifier even at strong and medium acidic conditions. [es

  7. Modelling and operation of reactors for enzymatic biodiesel production

    DEFF Research Database (Denmark)

    Price, Jason Anthony

    to the production of high fructose corn syrup, upgrading of fats and oils and biodiesel production to name a few. Despite these examples of industrial enzymatic applications, it is still not “clear cut” how to implement biocatalyst in industry and how best to optimize the processes. This is because the processing...... aspects of the enzyme with reaction/reactor engineering is performed. This strategy is applied to a case study of biodiesel production catalysed by a liquid enzyme formulation. The use of enzymes for biodiesel production is still in its infancy with non-optimized process designs. Furthermore is it unclear...

  8. Enzymatic characterization of lipid-based drug delivery systems

    DEFF Research Database (Denmark)

    Ljusberg-Wahren, Helena; Seier Nielsen, Flemming; Brogård, Mattias

    2005-01-01

    The present work introduces a simple and robust in vitro method for enzymatic characterisation of surface properties of lipid dispersions in aqueous media. The initial lipolysis rate in biorelevant media, using pancreatic lipase and a self-microemulsifying formulation (SMEDDS) containing digestible...... lipids as substrate, was determined. The impact of incorporating two sparingly water soluble model drugs, probucol and halofantrine, into the SMEDDS was studied. It was found that both model drugs reduced the initial rate of lipolysis compared with the vehicle, probucol having a larger effect than...

  9. Enzymatic description of the anhydrofructose pathway of glycogen degradation. I

    DEFF Research Database (Denmark)

    Yu, Shukun; Refdahl, Charlotte; Lundt, Inge

    2004-01-01

    The anhydrofructose pathway describes the degradation of glycogen and starch to metabolites via 1,5-anhydro-D-fructose (1,5AnFru). The enzyme catalyzing the first reaction step of this pathway, i.e., a-1,4-glucan lyase (EC 4.2.1.13), has been purified, cloned and characterized from fungi and red...... possessed all enzymes needed for conversion of glycogen to APP, an a-1,4-glucan lyase from this fungus was isolated and partially sequenced. Based on this work, a scheme of the enzymatic description of the anhydrofructose pathway in A. melaloma was proposed. Keywords: Anhydrofructose pathway; Anthracobia...

  10. Benefits from additives and xylanase during enzymatic hydrolysis of bamboo shoot and mature bamboo.

    Science.gov (United States)

    Li, Kena; Wang, Xiao; Wang, Jingfeng; Zhang, Junhua

    2015-09-01

    Effects of additives (BSA, PEG 6000, and Tween 80) on enzymatic hydrolysis of bamboo shoot and mature bamboo fractions (bamboo green, bamboo timber, bamboo yellow, bamboo node, and bamboo branches) by cellulases and/or xylanase were evaluated. The addition of additives was comparable to the increase of cellulase loadings in the conversion of cellulose and xylan in bamboo fractions. Supplementation of xylanase (1 mg/g DM) with cellulases (10 FPU/g DM) in the hydrolysis of bamboo fractions was more efficient than addition of additives in the production of glucose and xylose. Moreover, addition of additives could further increase the glucose release from different bamboo fractions by cellulases and xylanase. Bamboo green exhibited the lowest hydrolyzability. Almost all of the polysaccharides in pretreated bamboo shoot fractions were hydrolyzed by cellulases with the addition of additives or xylanase. Additives and xylanase showed great potential for reducing cellulase requirement in the hydrolysis of bamboo. Copyright © 2015 Elsevier Ltd. All rights reserved.

  11. The effect of irradiation temperature on the non-enzymatic browning reaction in cooked rice

    International Nuclear Information System (INIS)

    Lee, Ju-Woon; Oh, Sang-Hee; Kim, Jae-Hun; Byun, Eui-Hong; Ree Kim, Mee; Baek, Min; Byun, Myung-Woo

    2007-01-01

    The effect of irradiation temperature on the non-enzymatic browning reaction in a sugar-glycine solution and cooked rice generated by gamma irradiation was evaluated in the present study. When the sugar-glycine solution and cooked rice were irradiated at room temperature, the browning reaction was dramatically increased during the post-irradiation period. In the case of irradiation at below the freezing point, the browning by irradiation was retarded during not only irradiation but also a post-irradiation period. The changes of the sugar profile, such as a sugar loss or reducing power of the irradiated sugar-glycine solution and the electron spin resonance signal intensity of the irradiated cooked rice were also decreased with lower irradiation temperature. The present results may suggest that the production of free radicals and a radiolysis product is inhibited during gamma irradiation in the frozen state and it may prevent the browning reaction generated by gamma irradiation from occurring

  12. The effect of irradiation temperature on the non-enzymatic browning reaction in cooked rice

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Ju-Woon [Radiation Application Research Division, Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute, PO Box 1266, Jeongeup, Jeonbuk 580-185 (Korea, Republic of); Oh, Sang-Hee [Radiation Application Research Division, Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute, PO Box 1266, Jeongeup, Jeonbuk 580-185 (Korea, Republic of); Kim, Jae-Hun [Radiation Application Research Division, Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute, PO Box 1266, Jeongeup, Jeonbuk 580-185 (Korea, Republic of); Byun, Eui-Hong [Radiation Application Research Division, Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute, PO Box 1266, Jeongeup, Jeonbuk 580-185 (Korea, Republic of); Ree Kim, Mee [Department of Food and Nutrition, Chungnam National University, Gung-Dong 220, Yuseong, Daejeon 305-764 (Korea, Republic of); Baek, Min [Atomic Energy Policy Division, Ministry of Science and Technology, Government Complex-Gwacheon, Kyunggi 427-715 (Korea, Republic of); Byun, Myung-Woo [Radiation Application Research Division, Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute, PO Box 1266, Jeongeup, Jeonbuk 580-185 (Korea, Republic of)]. E-mail: mwbyun@kaeri.re.kr

    2007-05-15

    The effect of irradiation temperature on the non-enzymatic browning reaction in a sugar-glycine solution and cooked rice generated by gamma irradiation was evaluated in the present study. When the sugar-glycine solution and cooked rice were irradiated at room temperature, the browning reaction was dramatically increased during the post-irradiation period. In the case of irradiation at below the freezing point, the browning by irradiation was retarded during not only irradiation but also a post-irradiation period. The changes of the sugar profile, such as a sugar loss or reducing power of the irradiated sugar-glycine solution and the electron spin resonance signal intensity of the irradiated cooked rice were also decreased with lower irradiation temperature. The present results may suggest that the production of free radicals and a radiolysis product is inhibited during gamma irradiation in the frozen state and it may prevent the browning reaction generated by gamma irradiation from occurring.

  13. Kinetic study on the inhibition of xanthine oxidase by acylated derivatives of flavonoids synthesised enzymatically.

    Science.gov (United States)

    de Araújo, Maria Elisa Melo Branco; Franco, Yollanda Edwirges Moreira; Alberto, Thiago Grando; Messias, Marcia Cristina Fernandes; Leme, Camila Wielewski; Sawaya, Alexandra Christine Helena Frankland; Carvalho, Patricia de Oliveira

    2017-12-01

    Studies have reported that flavonoids inhibit xanthine oxidase (XO) activity; however, poor solubility and stability in lipophilic media limit their bioavailability and applications. This study evaluated the kinetic parameters of XO inhibition and partition coefficients of flavonoid esters biosynthesised from hesperidin, naringin, and rutin via enzymatic acylation with hexanoic, octanoic, decanoic, lauric, and oleic acids catalysed by Candida antarctica lipase B (CALB). Quantitative determination by ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS) showed higher conversion yields (%) for naringin and rutin esters using acyl donors with 8C and 10C. Rutin decanoate had higher partition coefficients (0.95), and naringin octanoate and naringin decanoate showed greater inhibitory effects on XO (IC 50 of 110.35 and 117.51 μM, respectively). Kinetic analysis showed significant differences (p flavonoids before and after acylation regarding K m values, whereas the values for V max were the same, implying the competitive nature of XO inhibition.

  14. Optimizing the Isolation of Microfibrillated Bamboo in High Pressure Enzymatic Hydrolysis

    Directory of Open Access Journals (Sweden)

    N. A. Sri Aprilia

    2015-07-01

    Full Text Available Bleached bamboo fiber was treated with a high pressure enzymatic hydrolysis (HPEH process in order to produce microfibrillated bamboo fiber (MBF. Mixture design of experiments was utilized to determine the optimal constituents of fiber, enzymes, and water for the HPEH process on the isolation yield of the MBF. Results showed the optimal combination for the maximal yield isolation of the MBF was 1 g fiber, 1 g enzyme, and 1 L water at 90 MPa and 70 °C. The influence of the reaction time of the HPEH process (6 to 48 h was also evaluated in this study. Morphological and thermal property analyses of untreated and treated bamboo fibers revealed that the HPEH process was effective for removing non-cellulosic components from the fibers. Thus, the HPEH process is an effective method for the isolation of the MBF, with the benefits of elevated crystallinity and thermal stability.

  15. Enzymatic Transesterification of Ethyl Ferulate with Fish Oil and Its Optimization by Response Surface Methodology

    DEFF Research Database (Denmark)

    Yang, Zhiyong; Glasius, Marianne; Xu, Xuebing

    2012-01-01

    formation of feruloyl fish oil products as well when appropriate amount of glycerol was present in the reaction. Therefore, the addition of equivalent molar amount of glycerol to EF was decided for the practical optimization of the system. The mutual effects of temperature (40 to 70 oC), reaction time (1......The enzymatic transesterification of ethyl ferulate (EF) with cod liver fish oil was investigated with Novozym 435 as catalyst under solvent-free conditions. The purpose of the study is to evaluate the synthesis system for production of feruloyl fish oil in industry. The modified HPLC method...... to 5 days), enzyme load (2 to 20 %) and substrate amount ratio of fish oil/EF (1 to 5) were thus studied with assistance of response surface methodology (RSM) for the purpose of maximizing the formation towards feruloyl fish oil. The models were well fitted and verified. The optimized conditions were...

  16. Improvement in Limit of Detection of Enzymatic Biogas Sensor Utilizing Chromatography Paper for Breath Analysis.

    Science.gov (United States)

    Motooka, Masanobu; Uno, Shigeyasu

    2018-02-02

    Breath analysis is considered to be an effective method for point-of-care diagnosis due to its noninvasiveness, quickness and simplicity. Gas sensors for breath analysis require detection of low-concentration substances. In this paper, we propose that reduction of the background current improves the limit of detection of enzymatic biogas sensors utilizing chromatography paper. After clarifying the cause of the background current, we reduced the background current by improving the fabrication process of the sensors utilizing paper. Finally, we evaluated the limit of detection of the sensor with the sample vapor of ethanol gas. The experiment showed about a 50% reduction of the limit of detection compared to previously-reported sensor. This result presents the possibility of the sensor being applied in diagnosis, such as for diabetes, by further lowering the limit of detection.

  17. Efficiency of new fungal cellulase systems in boosting enzymatic degradation of barley straw lignocellulose

    DEFF Research Database (Denmark)

    Rosgaard, L.; Pedersen, S.; Meyer, Anne Boye Strunge

    2006-01-01

    This study examined the cellulytic effects on steam-pretreated barley straw of cellulose-degrading enzyme systems from the five thermophilic fungi Chaetomium thermophilum, Thielavia terrestris, Thermoascus aurantiacus, Corynascus thermophilus, and Myceliophthora thermophila and from the mesophile...... Penicillum funiculosum. The catalytic glucose release was compared after treatments with each of the crude enzyme systems when added to a benchmark blend of a commercial cellulase product, Celluclast, derived from Trichoderma reesei and a P-glucosidase, Novozym 188, from Aspergillus niger. The enzymatic...... treatments were evaluated in an experimental design template comprising a span of pH (3.5-6.5) and temperature (35-65 degrees C) reaction combinations. The addition to Celluclast + Novozym 188 of low dosages of the crude enzyme systems, corresponding to 10 wt % of the total enzyme protein load, increased...

  18. Enzymatic depolymerization of gum Tragacanth: Bifidogenic potential of low molecular weight oligosaccharides

    DEFF Research Database (Denmark)

    Ahmadi Gavlighi, Hassan; Michalak, Malwina; Meyer, Anne S.

    2013-01-01

    Gum tragacanth derived from the plant “goat’s horn” (Astragalus sp.) has a long history of use as a stabilizing, viscosity-enhancing agent in food emulsions. The gum contains pectinaceous arabinogalactans and fucose-substituted xylogalacturonans. In this work, gum tragacanth from Astragalus...... and galactose content. The growth-stimulating potential of the three enzymatically produced gum tragacanth fractions was evaluated via growth assessment on seven different probiotic strains in single culture fermentations on: Bifidobacterium longum subsp. longum (2 strains), B. longum subsp. infantis (3 strains...... that on galactan (control). HAG3 completely inhibited the growth of the Cl. perfringens strain. Tragacanth gum is thus a potential source of prebiotic carbohydrates that exert no viscosity effects and which may find use as natural functional food ingredients....

  19. Improvement on sugar cane bagasse hydrolysis using enzymatic mixture designed cocktail.

    Science.gov (United States)

    Bussamra, Bianca Consorti; Freitas, Sindelia; Costa, Aline Carvalho da

    2015-01-01

    The aim of this work was to study cocktail supplementation for sugar cane bagasse hydrolysis, where the enzymes were provided from both commercial source and microorganism cultivation (Trichoderma reesei and genetically modified Escherichia coli), followed by purification. Experimental simplex lattice mixture design was performed to optimize the enzymatic proportion. The response was evaluated through hydrolysis microassays validated here. The optimized enzyme mixture, comprised of T. reesei fraction (80%), endoglucanase (10%) and β-glucosidase (10%), converted, theoretically, 72% of cellulose present in hydrothermally pretreated bagasse, whereas commercial Celluclast 1.5L converts 49.11%±0.49. Thus, a rational enzyme mixture designed by using synergism concept and statistical analysis was capable of improving biomass saccharification. Copyright © 2015 Elsevier Ltd. All rights reserved.

  20. Electron beam application as pre treatment of sugar cane bagasse to enzymatic hydrolysis of cellulose

    International Nuclear Information System (INIS)

    Cardoso, Vanessa Miguel

    2008-01-01

    Due to increasing worldwide shortage of food and energy sources, sugarcane bagasse has been considered as a substrate for single cell protein, animal feed, and renewable energy production. Sugarcane bagasse generally contain up to 45% glucose polymer cellulose, much of which is in a crystalline structure, 40% hemicelluloses, an amorphous polymer usually composed of xylose, arabinose, galactose, glucose, and mannose and 20% lignin, which cannot be easily separated into readily usable components due to their recalcitrant nature. Pure cellulose is readily depolymerised by radiation, but in biomass the cellulose is intimately bonded with lignin, that protect it from radiation effects. The objective of this study was the evaluation of the electron beam irradiation efficiency as a pre-treatment to enzymatic hydrolysis of cellulose in order to facilitate its fermentation and improves the production of ethanol biofuel. Samples of sugarcane bagasse were obtained in sugar/ethanol Mill sited in Piracicaba, Brazil, and were irradiated using Radiation Dynamics Electron Beam Accelerator with 1,5 MeV energy and 37 kW, in batch systems. The applied absorbed doses of the fist sampling, Bagasse A, were 20 kGy, 50 kGy, 10 0 kGy and 200 kGy. After the evaluation the preliminary obtained results, it was applied lower absorbed doses in the second assay: 5 kGy, 10 kGy, 20 kGy, 30 kGy, 50 kGy, 70 kGy, 100 kGy and 150 kGy. The electron beam processing took to changes in the sugarcane bagasse structure and composition, lignin and cellulose cleavage. The yield of enzymatic hydrolyzes of cellulose in. (author)

  1. High Hydrostatic Pressure-Assisted Enzymatic Treatment Improves Antioxidant and Anti-inflammatory Properties of Phosvitin.

    Science.gov (United States)

    Yoo, Heejoo; Bamdad, Fatemeh; Gujral, Naiyana; Suh, Joo-Won; Sunwoo, Hoon

    2017-01-01

    Phosvitin (PV) is a highly-phosphorylated metal-binding protein in egg yolk. Phosphoserine clusters make PV resistant to enzymatic digestion, which might be nutritionally undesirable. This study was designed to determine the effects of high hydrostatic pressure and enzymatic hydrolysis (HHP-EH) on the antioxidant and anti-inflammatory properties of PV hydrolysates (PVHs). PV was hydrolyzed by alcalase, elastase, savinase, thermolysin, and trypsin at 0.1, 50, and 100 MPa pressure levels. PVHs were evaluated for degree of hydrolysis, molecular weight distribution patterns, antioxidant and anti-inflammatory properties in chemical and cellular models. The effect of PVH on gene expression of pro-inflammatory cytokines (TNF-α and IL-1β) was also evaluated using real time-PCR. The hydrolysate with most potent antioxidant and anti-inflammatory properties was subjected to LC-MS/MS analysis to identify the peptide sequence. Hydrolysates produced at 100 MPa exhibited higher degree of hydrolysis and greater reducing power and free radical scavenging activity compared to those obtained at atmospheric pressure. After adjusting the phosphate content, alcalase- and trypsin-digested PVHs showed superior iron chelation capacity (69-73%), regardless of pressure. Both alcalase- and trypsin-digested PVHs significantly inhibited nitric oxide production by RAW264.7 macrophage cells. LPS-stimulated up-regulation of proinflammatory cytokines was also suppressed by alcalase-digested PVH. The HHP-EH method could play a promising role in the production of bioactive peptides from hydrolysis-resistant proteins. HHP-assisted PVH may be useful in preparing a potential pharmaceutical with antioxidant and anti-inflammatory properties. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  2. Paper-based maskless enzymatic sensor for glucose determination combining ink and wire electrodes.

    Science.gov (United States)

    Amor-Gutiérrez, O; Costa Rama, E; Costa-García, A; Fernández-Abedul, M T

    2017-07-15

    In this work we have developed an amperometric enzymatic biosensor in a paper-based platform with a mixed electrode configuration: carbon ink for the working electrode (WE) and metal wires (from a low-cost standard electronic connection) for reference (RE) and auxiliary electrodes (AE). A hydrophobic wax-defined paper area was impregnated with diluted carbon ink. Three gold-plated pins of the standard connection are employed, one for connecting the WE and the other two acting as RE and AE. The standard connection works as a clip in order to support the paper in between. As a proof-of-concept, glucose sensing was evaluated. The enzyme cocktail (glucose oxidase, horseradish peroxidase and potassium ferrocyanide as mediator of the electron transfer) was adsorbed on the surface. After drying, glucose solution was added to the paper, on the opposite side of the carbon ink. It wets RE and AE, and flows by capillarity through the paper contacting the carbon WE surface. The reduction current of ferricyanide, product of the enzymatic reaction, is measured chronoamperometrically and correlates to the concentration of glucose. Different parameters related to the bioassay were optimized, adhering the piece of paper onto a conventional screen-printed carbon electrode (SPCE). In this way, the RE and the AE of the commercial card were employed for optimizing the paper-WE. After evaluating the assay system in the hybrid paper-SPCE cell, the three-electrode system consisting of paper-WE, wire-RE and wire-AE, was employed for glucose determination, achieving a linear range between 0.3 and 15mM with good analytical features and being able of quantifying glucose in real food samples. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. Enzymatic cybernetics: an unpublished work by Jacques Monod.

    Science.gov (United States)

    Gayon, Jean

    2015-06-01

    In 1959, Jacques Monod wrote a manuscript entitled Cybernétique enzymatique [Enzymatic cybernetics]. Never published, this unpublished manuscript presents a synthesis of how Monod interpreted enzymatic adaptation just before the publication of the famous papers of the 1960s on the operon. In addition, Monod offers an example of a philosophy of biology immersed in scientific investigation. Monod's philosophical thoughts are classified into two categories, methodological and ontological. On the methodological side, Monod explicitly hints at his preferences regarding the scientific method in general: hypothetical-deductive method, and use of theoretical models. He also makes heuristic proposals regarding molecular biology: the need to analyse the phenomena in question at the level of individual cells, and the dual aspect of all biological explanation, functional and evolutionary. Ontological issues deal with the notions of information and genetic determinism, "cellular memory", the irrelevance of the notion of "living matter", and the usefulness of a cybernetic comprehension of molecular biology. Copyright © 2015 Académie des sciences. Published by Elsevier SAS. All rights reserved.

  4. Enzymatic Digestion of Chronic Wasting Disease Prions Bound to Soil

    Science.gov (United States)

    SAUNDERS, SAMUEL E.; BARTZ, JASON C.; VERCAUTEREN, KURT C.; BARTELT-HUNT, SHANNON L.

    2010-01-01

    Chronic wasting disease (CWD) and sheep scrapie can be transmitted via indirect environmental routes, and it is known that soil can serve as a reservoir of prion infectivity. Given the strong interaction between the prion protein (PrP) and soil, we hypothesized that binding to soil enhances prion resistance to enzymatic digestion, thereby facilitating prion longevity in the environment and providing protection from host degradation. We characterized the performance of a commercially available subtilisin enzyme, the Prionzyme, to degrade soil-bound and unbound CWD and HY TME PrP as a function of pH, temperature, and treatment time. The subtilisin enzyme effectively degraded PrP adsorbed to a wide range of soils and soil minerals below the limits of detection. Signal loss occurred rapidly at high pH (12.5) and within 7 d under conditions representative of the natural environment (pH 7.4, 22°C). We observed no apparent difference in enzyme effectiveness between bound and unbound CWD PrP. Our results show that although adsorbed prions do retain relative resistance to enzymatic digestion compared with other brain homogenate proteins, they can be effectively degraded when bound to soil. Our results also suggest a topical application of a subtilisin enzyme solution may be an effective decontamination method to limit disease transmission via environmental ‘hot spots’ of prion infectivity. PMID:20450190

  5. Cellulase-lignin interactions in the enzymatic hydrolysis of lignocellulose

    Energy Technology Data Exchange (ETDEWEB)

    Rahikainen, J.

    2013-11-01

    Today, the production of transportation fuels and chemicals is heavily dependent on fossil carbon sources, such as oil and natural gas. Their limited availability and the environmental concerns arising from their use have driven the search for renewable alternatives. Lignocellulosic plant biomass is the most abundant, but currently underutilised, renewable carbon-rich resource for fuel and chemical production. Enzymatic degradation of structural polysaccharides in lignocellulose produces soluble carbohydrates that serve as ideal precursors for the production of a vast amount of different chemical compounds. The difficulty in full exploitation of lignocellulose for fuel and chemical production lies in the complex and recalcitrant structure of the raw material. Lignocellulose is mainly composed of structural polysaccharides, cellulose and hemicellulose, but also of lignin, which is an aromatic polymer. Enzymatic degradation of cellulose and hemicellulose is restricted by several substrate- and enzyme-related factors, among which lignin is considered as one of the most problematic issues. Lignin restricts the action of hydrolytic enzymes and enzyme binding onto lignin has been identified as a major inhibitory mechanism preventing efficient hydrolysis of lignocellulosic feedstocks. In this thesis, the interactions between cellulase enzymes and lignin-rich compounds were studied in detail and the findings reported in this work have the potential to help in controlling the harmful cellulase-lignin interactions, and thus improve the biochemical processing route from lignocellulose to fuels and chemicals.

  6. Chemical and Enzymatic Hydrolysis of Polyurethane/Polylactide Blends

    Directory of Open Access Journals (Sweden)

    Joanna Brzeska

    2015-01-01

    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.

  7. Evolutionary selection of enzymatically synthesized semiconductors from biomimetic mineralization vesicles.

    Science.gov (United States)

    Bawazer, Lukmaan A; Izumi, Michi; Kolodin, Dmitriy; Neilson, James R; Schwenzer, Birgit; Morse, Daniel E

    2012-06-26

    The way nature evolves and sculpts materials using proteins inspires new approaches to materials engineering but is still not completely understood. Here, we present a cell-free synthetic biological platform to advance studies of biologically synthesized solid-state materials. This platform is capable of simultaneously exerting many of the hierarchical levels of control found in natural biomineralization, including genetic, chemical, spatial, structural, and morphological control, while supporting the evolutionary selection of new mineralizing proteins and the corresponding genetically encoded materials that they produce. DNA-directed protein expression and enzymatic mineralization occur on polystyrene microbeads in water-in-oil emulsions, yielding synthetic surrogates of biomineralizing cells that are then screened by flow sorting, with light-scattering signals used to sort the resulting mineralized composites differentially. We demonstrate the utility of this platform by evolutionarily selecting newly identified silicateins, biomineralizing enzymes previously identified from the silica skeleton of a marine sponge, for enzyme variants capable of synthesizing silicon dioxide (silica) or titanium dioxide (titania) composites. Mineral composites of intermediate strength are preferentially selected to remain intact for identification during cell sorting, and then to collapse postsorting to expose the encoding genes for enzymatic DNA amplification. Some of the newly selected silicatein variants catalyze the formation of crystalline silicates, whereas the parent silicateins lack this ability. The demonstrated bioengineered route to previously undescribed materials introduces in vitro enzyme selection as a viable strategy for mimicking genetic evolution of materials as it occurs in nature.

  8. PARP1 Val762Ala polymorphism reduces enzymatic activity

    International Nuclear Information System (INIS)

    Wang Xiaogan; Wang Zhaoqi; Tong Weimin; Shen Yan

    2007-01-01

    Poly(ADP-ribose) polymerase 1 (PARP1) modifies a variety of nuclear proteins by poly(ADP-ribosyl)ation, and plays diverse roles in molecular and cellular processes. A common PARP1 single nucleotide polymorphism (SNP) at codon 762, resulting in the substitution of alanine (Ala) for valine (Val) in the catalytic domain has been implicated in susceptibility to cancer. To characterize the functional effect of this polymorphism on PARP1, we performed in vitro enzymatic analysis on PARP1-Ala762 and PARP1-Val762. We found that PARP1-Ala762 displayed 57.2% of the activity of PARP1-Val762 for auto-poly(ADP-ribosyl)ation and 61.9% of the activity of PARP1-Val762 for trans-poly(ADP-ribosyl)ation of histone H1. The kinetic characterization revealed that the K m of PARP1-Ala762 was increased to a 1.2-fold of the K m of PARP1-Val762 for trans-poly(ADP-ribosyl)ation. Thus, the PARP1 Val762Ala polymorphism reduces the enzymatic activity of PARP1 by increasing K m . This finding suggests that different levels of poly(ADP-ribosyl)ation by PARP1 might aid in understanding Cancer risk of carriers of the PARP1 Val762Ala polymorphism

  9. A characterization of scale invariant responses in enzymatic networks.

    Directory of Open Access Journals (Sweden)

    Maja Skataric

    Full Text Available An ubiquitous property of biological sensory systems is adaptation: a step increase in stimulus triggers an initial change in a biochemical or physiological response, followed by a more gradual relaxation toward a basal, pre-stimulus level. Adaptation helps maintain essential variables within acceptable bounds and allows organisms to readjust themselves to an optimum and non-saturating sensitivity range when faced with a prolonged change in their environment. Recently, it was shown theoretically and experimentally that many adapting systems, both at the organism and single-cell level, enjoy a remarkable additional feature: scale invariance, meaning that the initial, transient behavior remains (approximately the same even when the background signal level is scaled. In this work, we set out to investigate under what conditions a broadly used model of biochemical enzymatic networks will exhibit scale-invariant behavior. An exhaustive computational study led us to discover a new property of surprising simplicity and generality, uniform linearizations with fast output (ULFO, whose validity we show is both necessary and sufficient for scale invariance of three-node enzymatic networks (and sufficient for any number of nodes. Based on this study, we go on to develop a mathematical explanation of how ULFO results in scale invariance. Our work provides a surprisingly consistent, simple, and general framework for understanding this phenomenon, and results in concrete experimental predictions.

  10. Effect of irradiation on enzymatic digestion of cellulosic wastes

    International Nuclear Information System (INIS)

    Chowdhury, N.A.; Matsuhashi, Shinpei; Hashimoto, Shoji; Kume, Tamikazu.

    1993-03-01

    Combination treatments with irradiation and other methods were examined to enhance the digestion of cellulosic materials such as sugar cane bagasse and rice straw. The amount of crude fiber (CF), acid detergent fiber (ADF) and neutral detergent fiber (NDF) of bagasse and rice straw were changed with various treatments. Alkali treatment (0.2N NaOH) was the most efficient for the enzymatic hydrolysis of bagasse and rice straw. Combination treatments with radiation and alkali or other methods increased their efficiency, and synergistic effect of radiation and alkali treatment was observed. Enzymatic digestion of CF of bagasse and rice straw treated by degassed water yielded high reducing sugar comparable to that of CF treated by alkali. CF of bagasse and rice straw treated by ozone did not show the significant increase in the release of reducing sugar upon saccharification. ADF and acid detergent lignin (ADL) contents decreased with the fermentation of bagasse by Coriolus versicolor. Electron microscopic observations also revealed the degradation of lignocellulosic components of bagasse. (author)

  11. Enzymatic U(VI) reduction by Desulfosporosinus species

    International Nuclear Information System (INIS)

    Suzuki, Y.; Kelly, S.D.; Kemner, K.M.; Banfield, J.F.

    2004-01-01

    Here we tested U(VI) reduction by a Desulfosporosinus species (sp.) isolate and type strain (DSM 765) in cell suspensions (pH 7) containing 1 mM U(VI) and lactate, under an atmosphere containing N 2 -CO 2 -H 2 (90: 5: 5). Although neither Desulfosporosinus species (spp.) reduced U(VI) in cell suspensions with 0.25% Na-bicarbonate or 0.85% NaCl, U(VI) was reduced in these solutions by a control strain, desulfovibrio desulfuricans (ATCC 642). However, both Desulfosporosinus strains reduced U(VI) in cell suspensions depleted in bicarbonate and NaCl. No U(VI) reduction was observed without lactate and H 2 electron donors or with heat-killed cells, indicating enzymatic U(VI) reduction. Uranium(VI) reduction by both strains was inhibited when 1 mM CuCl 2 was added to the cell suspensions. Because the Desulfosporosinus DSM 765 does not contain cytochrome c 3 used by Desulfovibrio spp. to reduce U(VI), Desulfosporosinus species reduce uranium via a different enzymatic pathway. (orig.)

  12. Applications of immobilized Thermomyces lanuginosa lipase in interesterification

    DEFF Research Database (Denmark)

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

    2003-01-01

    (RSM). Thermomyces lanuginosa lipase had an activity similar to that of immobilized Rhizomucor miehei lipase (Lipozyme RM IM) in the glycerolysis of sunflower oil, but the former had higher activity at a low reaction temperature (5degreesC). Thermomyces lanuginosa lipase was found to have much lower...... catalytic activity than Lipozyme RM IM in the acidolysis of sunflower oil with caprylic acid. However, the activity of T. lanuginosa lipase was only slightly lower than that of Lipozyme RM IM in the ester-ester exchange between tripalmitin (PPP) and the ethyl esters of EPA and DHA (EE). For this reason...

  13. Enzymatic modification of natural and synthetic polymers using lipases and proteases

    Science.gov (United States)

    Chakraborty, Soma

    corresponding copolymers. As an alternative to chemical hydrolysis a mild and selective enzymatic method was discovered. Fifteen proteases were evaluated for this transformation. Of these, PROT 7 was the most active. Within 24h PROT 7 gave products with 44% hydrolysis. Further hydrolysis was not observed by extending the reaction time because poly(vinylformamide-co-40%vinylamine) is a poor substrate for further hydrolysis. The sequence distribution of copolymers formed by chemical hydrolysis and enzymatic hydrolysis was compared. Chemical hydrolysis gave random copolymer. In contrast, PROT 7 gave block-like arrangement of VAm units.

  14. The enzymatic and antioxidative stress response of Lemna minor to copper and a chloroacetamide herbicide.

    Science.gov (United States)

    Obermeier, Michael; Schröder, Christian A; Helmreich, Brigitte; Schröder, Peter

    2015-12-01

    Lemna minor L., a widely used model plant for toxicity tests has raised interest for its application to phytoremediation due to its rapid growth and ubiquitous occurrence. In rural areas, the pollution of water bodies with heavy metals and agrochemicals poses a problem to surface water quality. Among problematic compounds, heavy metals (copper) and pesticides are frequently found in water bodies. To establish duckweed as a potential plant for phytoremediation, enzymatic and antioxidative stress responses of Lemna minor during exposure to copper and a chloroacetamide herbicide were investigated in laboratory studies. The present study aimed at evaluating growth and the antioxidative and glutathione-dependent enzyme activity of Lemna plants and its performance in a scenario for phytoremediation of copper and a chloroacetamide herbicide. Lemna minor was grown in Steinberg medium under controlled conditions. Plants were treated with CuSO4 (ion conc. 50 and 100 μg/L) and pethoxamide (1.25 and 2.5 μg/L). Measurements following published methods focused on plant growth, oxidative stress, and basic detoxification enzymes. Duckweed proved to survive treatment with the respective concentrations of both pollutants very well. Its growth was inhibited scarcely, and no visible symptoms occurred. On the cellular basis, accumulation of O2(-) and H2O2 were detected, as well as stress reactions of antioxidative enzymes. Duckweed detoxification potential for organic pollutants was high and increased significantly with incubation. Pethoxamide was found to be conjugated with glutathione. Copper was accumulated in the fronds at high levels, and transient oxidative defense reactions were triggered. This work confirms the significance of L. minor for the removal of copper from water and the conjugation of the selective herbicide pethoxamide. Both organic and inorganic xenobiotics induced different trends of enzymatic and antioxidative stress response. The strong increase of stress

  15. Chemical composition and enzymatic digestibility of sugarcane clones selected for varied lignin content

    Directory of Open Access Journals (Sweden)

    Masarin Fernando

    2011-12-01

    Full Text Available Abstract Background The recalcitrance of lignocellulosic materials is a major limitation for their conversion into fermentable sugars. Lignin depletion in new cultivars or transgenic plants has been identified as a way to diminish this recalcitrance. In this study, we assessed the success of a sugarcane breeding program in selecting sugarcane plants with low lignin content, and report the chemical composition and agronomic characteristics of eleven experimental hybrids and two reference samples. The enzymatic digestion of untreated and chemically delignified samples was evaluated to advance the performance of the sugarcane residue (bagasse in cellulosic-ethanol production processes. Results The ranges for the percentages of glucan, hemicellulose, lignin, and extractive (based on oven-dry biomass of the experimental hybrids and reference samples were 38% to 43%, 25% to 32%, 17% to 24%, and 1.6% to 7.5%, respectively. The samples with the smallest amounts of lignin did not produce the largest amounts of total polysaccharides. Instead, a variable increase in the mass of a number of components, including extractives, seemed to compensate for the reduction in lignin content. Hydroxycinnamic acids accounted for a significant part of the aromatic compounds in the samples, with p-coumaric acid predominating, whereas ferulic acid was present only in low amounts. Hydroxycinnamic acids with ester linkage to the hemicelluloses varied from 2.3% to 3.6%. The percentage of total hydroxycinnamic acids (including the fraction linked to lignin through ether linkages varied from 5.0% to 9.2%, and correlated to some extent with the lignin content. These clones released up to 31% of glucose after 72 hours of digestion with commercial cellulases, whereas chemically delignified samples led to cellulose conversion values of more than 80%. However, plants with lower lignin content required less delignification to reach higher efficiencies of cellulose conversion during

  16. Enzymatic Xylose Release from Pretreated Corn Bran Arabinoxylan: Differential Effects of Deacetylation and Deferuloylation on Insoluble and Soluble Substrate Fractions

    DEFF Research Database (Denmark)

    Agger, Jane; Viksø-Nielsen, Ander; Meyer, Anne S.

    2010-01-01

    In the present work enzymatic hydrolysis of arabinoxylan from pretreated corn bran (190 °C, 10 min) was evaluated by measuring the release of xylose and arabinose after treatment with a designed minimal mixture of monocomponent enzymes consisting of α-l-arabinofuranosidases, an endoxylanase......, and a β-xylosidase. The pretreatment divided the corn bran material 50:50 into soluble and insoluble fractions having A:X ratios of 0.66 and 0.40, respectively. Addition of acetyl xylan esterase to the monocomponent enzyme mixture almost doubled the xylose release from the insoluble substrate fraction...

  17. Optimization of reaction parameters for enzymatic glyceride synthesis from fish oil: Ethyl esters versus free fatty acids

    DEFF Research Database (Denmark)

    Ravn, Helle Christine; Damstrup, Marianne L.; Meyer, Anne S.

    2012-01-01

    Enzymatic conversion of fish oil free fatty acids (FFA) or fatty acid ethyl esters (FAE) into glycerides via esterification or transesterification was examined. The reactions catalyzed by Lipozyme™ 435, a Candida antarctica lipase, were optimized. Influence on conversion yields of fatty acid chain...... length, saturation degree, temperature, enzyme dosage, molar ratio glycerol:fatty acids, acyl source composition (w/w ratio FFA:FAE), and reaction time was evaluated collectively by multiple linear regression. All reaction variables influenced the conversion into glycerides. Transesterification of FAE...

  18. Sequential enzymatic synthesis and separation of 13N-L-glutamic acid and 13N-L-alanine

    International Nuclear Information System (INIS)

    Cohen, M.B.; Spolter, L.; MacDonald, M.; Chang, C.C.; Takahashi, J.

    1975-01-01

    The sequential enzymatic synthesis and separation of 13 N-L-glutamic acid and 13 N-L-alanine are described. Basically, that involves the synthesis of 13 N-L-glutamic acid by one enzyme, the transamination of the labeled glutamic acid to form 13 N-L-alanine by a second enzyme, and the separation of the two amino acids by rapid column chromatography. The 13 N-L-alanine was evaluated in animals by imaging and tissue distribution studies and showed good potential as a pancreatic imaging agent

  19. Influence of Different Lignocellulose Sources on Endo-1,4-β-Glucanase Gene Expression and Enzymatic Activity of Bacillus amyloliquefaciens B31C

    Directory of Open Access Journals (Sweden)

    Rosangela Di Pasqua

    2014-01-01

    Full Text Available Conversion of cellulose into fermentable sugars for ethanol production is currently performed by enzymatic hydrolysis catalyzed by cellulases. The cellulases are produced by a wide variety of microorganisms, playing a major role in the recycling of biomass. The endo-1,4-β-glucanase (CelB31C from Bacillus amyloliquefaciens B31C, isolated from compost and previously selected on the basis of highest cellulase activity levels among Bacillus isolated, was characterized as being a potential candidate for a biocatalyst in lignocellulose conversion for second-generation bioethanol production. The aim of this work was to evaluate the changes in production of enzymatic activity of the endo-1,4-β-glucanase (CelB31C and the expression of its gene (bglC using a carboxymethylcellulase activity assay and qRT-PCR analysis, respectively, during growth of B. amyloliquefaciens B31C on different cellulose sources: carboxymethylcellulose (CMC, pure cellulose from Arundo donax, pretreated Arundo donax biomass (Chemtex, and microcrystalline cellulose (Avicel. The results showed that both the expression of bglC gene and the enzymatic activity production are related to the type of cellulose source. The strain showed a high enzymatic activity on lignocellulosic biomass and on microcrystalline cellulose. Furthermore, the highest gene expression occurred during the exponential phase of growth, except in the presence of Avicel.

  20. Effect of soya lecithin on the enzymatic system of the white-rot fungi Anthracophyllum discolor.

    Science.gov (United States)

    Bustamante, M; González, M E; Cartes, A; Diez, M C

    2011-01-01

    The present work optimized the initial pH of the medium and the incubation temperature for ligninolytic enzymes produced by the white-rot fungus Anthracophyllum discolor. Additionally, the effect of soya lecithin on mycelial growth and the production of ligninolytic enzymes in static batch cultures were evaluated. The critical micelle concentration of soya lecithin was also studied by conductivity. The effects of the initial pH (3, 4, and 5) and incubation temperature (20, 25, and 30°C) on different enzymatic activities revealed that the optimum conditions to maximize ligninolytic activity were 26°C and pH 5.5 for laccase and manganese peroxidase (MnP) and 30°C and pH 5.5 for manganese-independent peroxidase (MiP). Under these culture conditions, the maximum enzyme production was 10.16, 484.46, and 112.50 U L(-1) for laccase, MnP, and manganese-independent peroxidase MiP, respectively. During the study of the effect of soya lecithin on A. discolor, we found that the increase in soya lecithin concentration from 0 to 10 g L(-1) caused an increase in mycelial growth. On the other hand, in the presence of soya lecithin, A. discolor produced mainly MnP, which reached a maximum concentration of 30.64 ± 4.61 U L(-1) after 25 days of incubation with 1 g L(-1) of the surfactant. The other enzymes were produced but to a lesser extent. The enzymatic activity of A. discolor was decreased when Tween 80 was used as a surfactant. The critical micelle concentration of soya lecithin calculated in our study was 0.61 g L(-1).

  1. Epigallocatechin gallate incorporation into lignin enhances the alkaline delignification and enzymatic saccharification of cell walls

    Directory of Open Access Journals (Sweden)

    Elumalai Sasikumar

    2012-08-01

    Full Text Available Abstract Background Lignin is an integral component of the plant cell wall matrix but impedes the conversion of biomass into biofuels. The plasticity of lignin biosynthesis should permit the inclusion of new compatible phenolic monomers such as flavonoids into cell wall lignins that are consequently less recalcitrant to biomass processing. In the present study, epigallocatechin gallate (EGCG was evaluated as a potential lignin bioengineering target for rendering biomass more amenable to processing for biofuel production. Results In vitro peroxidase-catalyzed polymerization experiments revealed that both gallate and pyrogallyl (B-ring moieties in EGCG underwent radical cross-coupling with monolignols mainly by β–O–4-type cross-coupling, producing benzodioxane units following rearomatization reactions. Biomimetic lignification of maize cell walls with a 3:1 molar ratio of monolignols and EGCG permitted extensive alkaline delignification of cell walls (72 to 92% that far exceeded that for lignified controls (44 to 62%. Alkali-insoluble residues from EGCG-lignified walls yielded up to 34% more glucose and total sugars following enzymatic saccharification than lignified controls. Conclusions It was found that EGCG readily copolymerized with monolignols to become integrally cross-coupled into cell wall lignins, where it greatly enhanced alkaline delignification and subsequent enzymatic saccharification. Improved delignification may be attributed to internal trapping of quinone-methide intermediates to prevent benzyl ether cross-linking of lignin to structural polysaccharides during lignification, and to the cleavage of ester intra-unit linkages within EGCG during pretreatment. Overall, our results suggest that apoplastic deposition of EGCG for incorporation into lignin would be a promising plant genetic engineering target for improving the delignification and saccharification of biomass crops.

  2. [Effects of hot-NaOH pretreatment on Jerusalem artichoke stalk composition and subsequent enzymatic hydrolysis].

    Science.gov (United States)

    Wang, Qing; Qiu, Jingwen; Li, Yang; Shen, Fei

    2015-10-01

    In order to explore the possibility of Jerusalem artichoke stalk for bioenergy conversion, we analyzed the main composition of whole stalk, pitch, and core of the stalk. Meanwhile, these parts were pretreated with different NaOH concentrations at 121 degrees C. Afterwards, enzymatic hydrolysis was performed to evaluate the pretreatment efficiency. Jerusalem artichoke stalk was characterized by relatively high lignin content (32.0%) compared with traditional crop stalks. The total carbohydrate content was close to that of crop stalks, but with higher cellulose content (40.5%) and lower hemicellulose (19.6%) than those of traditional crop stalks. After pretreatment, the lignin content in the whole stalk, pitch, and core decreased by 13.1%-13.4%, 8.3%-13.5%, and 19.9%-27.2%, respectively, compared with the unpretreated substrates. The hemicellulose content in the whole stalk, pitch, and core decreased 87.8%-96.9%, 87.6%-95.0%, and 74.0%-90.2%, respectively. Correspondingly, the cellulose content in the pretreated whole stalk, pitch, and core increased by 56.5%-60.2%, 52.2%-55.4%, and 62.7%-73.2%, respectively. Moreover, increase of NaOH concentration for pretreatment could improve the enzymatic hydrolysis of the whole stalk and pitch by 2.3-2.6 folds and 10.3-18.5 folds, respectively. The hydrolysis of pretreated stalk core decreased significantly as 2.0 mol/L NaOH was employed, although the increased NaOH concentration can also improve its hydrolysis performance. Based on these results, hot-NaOH can be regarded as an option for Jerusalem artichoke stalk pretreatment. Increasing NaOH concentration was beneficial to hemicellulose and lignin removal, and consequently improved sugar conversion. However, the potential decrease of sugar conversion of the pretreated core by higher NaOH concentration suggested further optimization on the pretreatment conditions should be performed.

  3. Identification of odor volatile compounds and deodorization of Paphia undulata enzymatic hydrolysate

    Science.gov (United States)

    Chen, Deke; Chen, Xin; Chen, Hua; Cai, Bingna; Wan, Peng; Zhu, Xiaolian; Sun, Han; Sun, Huili; Pan, Jianyu

    2016-12-01

    Unfavorable fishy odour is an inevitable problem in aquatic products. In the present study, headspace solid-phase microextraction gas chromatography mass spectrometry (HS-SPME-GC-MS) analysis of volatiles from untreated samples and three deodorized samples (under the optimal conditions) of Paphia undulata enzymatic hydrolysate revealed that the compounds contributing to the distinctive odor were 1-octen-3-ol, n-hexanal, n-heptanal, 2,4-heptadienal, and 2,4-decadienal, whereas n-pentanal, n-octanal, n-octanol, benzaldehyde, 2-ethylfuran and 2-pentylfuran were the main contributors to the aromatic flavor. The deodorizing effects of activated carbon (AC) adsorption, yeast extract (YE) masking and tea polyphenol (TP) treatment on a P. undulata enzymatic hydrolysate were investigated using orthogonal experiments with sensory evaluation as the index. The following optimized deodorization conditions were obtained: AC adsorption (35 mg mL-1, 80°C, 40 min), YE masking (7 mg mL-1, 45°C, 30 min) and TP treatment (0.4 mg mL-1, 40°C, 50 min). AC adsorption effectively removed off-flavor volatile aldehydes and ketones. YE masking modified the odor profile by increasing the relative contents of aromatic compounds and decreasing the relative contents of aldehydes and ketones. The TP treatment was not effective in reducing the odor score, but it significantly reduced the relative content of aldehydes while increasing that of alkanes. It is also notable that TP effectively suppressed trimethylamine (TMA) formation in a P. undulate hydrolysate solution for a period of 72 h.

  4. Sol-Gel Derived Adsorbents with Enzymatic and Complexonate Functions for Complex Water Remediation

    Directory of Open Access Journals (Sweden)

    Roman P. Pogorilyi

    2017-09-01

    Full Text Available Sol-gel technology is a versatile tool for preparation of complex silica-based materials with targeting functions for use as adsorbents in water purification. Most efficient removal of organic pollutants is achieved by using enzymatic reagents grafted on nano-carriers. However, enzymes are easily deactivated in the presence of heavy metal cations. In this work, we avoided inactivation of immobilized urease by Cu (II and Cd (II ions using magnetic nanoparticles provided with additional complexonate (diethylene triamine pentaacetic acid or DTPA functions. Obtained nanomaterials were characterized by Fourier transform infrared spectroscopy (FTIR, thermogravimetric analysis (TGA, and scanning electron microscopy (SEM. According to TGA, the obtained Fe3O4/SiO2-NH2-DTPA nanoadsorbents contained up to 0.401 mmol/g of DTPA groups. In the concentration range Ceq = 0–50 mmol/L, maximum adsorption capacities towards Cu (II and Cd (II ions were 1.1 mmol/g and 1.7 mmol/g, respectively. Langmuir adsorption model fits experimental data in concentration range Ceq = 0–10 mmol/L. The adsorption mechanisms have been evaluated for both of cations. Crosslinking of 5 wt % of immobilized urease with glutaraldehyde prevented the loss of the enzyme in repeated use of the adsorbent and improved the stability of the enzymatic function leading to unchanged activity in at least 18 cycles. Crosslinking of 10 wt % urease on the surface of the particles allowed a decrease in urea concentration in 20 mmol/L model solutions to 2 mmol/L in up to 10 consequent decomposition cycles. Due to the presence of DTPA groups, Cu2+ ions in concentration 1 µmol/L did not significantly affect the urease activity. Obtained magnetic Fe3O4/SiO2-NH2-DTPA-Urease nanocomposite sorbents revealed a high potential for urease decomposition, even in presence of heavy metal ions.

  5. Enzymatic degradation of in vitro Staphylococcus aureus biofilms supplemented with human plasma

    Directory of Open Access Journals (Sweden)

    Watters CM

    2016-04-01

    Full Text Available Chase M Watters,1,2 Tarea Burton,1 Dickson K Kirui,1 Nancy J Millenbaugh1 1Maxillofacial Injury and Disease Department, Naval Medical Research Unit San Antonio, Joint Base San Antonio-Fort Sam Houston, TX, USA; 2Wound Infections Department, Naval Medical Research Center, Silver Spring, MD, USA Abstract: Enzymatic debridement is a therapeutic strategy used clinically to remove necrotic tissue from wounds. Some of the enzymes utilized for debridement have been tested against bacterial pathogens, but the effectiveness of these agents in dispersing clinically relevant biofilms has not been fully characterized. Here, we developed an in vitro Staphylococcus aureus biofilm model that mimics wound-like conditions and employed this model to investigate the antibiofilm activity of four enzymatic compounds. Human plasma at concentrations of 0%–50% was supplemented into growth media and used to evaluate biofilm biomass accumulation over 24 hours and 48 hours in one methicillin-sensitive and five methicillin-resistant strains of S. aureus. Supplementation of media with 10% human plasma resulted in the most robust biofilms in all six strains. The enzymes α-amylase, bromelain, lysostaphin, and papain were then tested against S. aureus biofilms cultured in 10% human plasma. Quantification of biofilms after 2 hours and 24 hours of treatment using the crystal violet assay revealed that lysostaphin decreased biomass by up to 76%, whereas a-amylase, bromelain, and papain reduced biomass by up to 97%, 98%, and 98%, respectively. Scanning electron microscopy confirmed that the dispersal agents detached the biofilm exopolysaccharide matrix and bacteria from the growth surface. Lysostaphin caused less visible dispersal of the biofilms, but unlike the other enzymes, induced morphological changes indicative of bacterial cell damage. Overall, our results indicate that use of enzymes may be an effective means of eradicating biofilms and a promising strategy to improve

  6. Enzymatic-fluorometric quantification of cholesterol in bovine milk

    DEFF Research Database (Denmark)

    Larsen, Torben

    2012-01-01

    The present paper describes an enzymatic–fluorometric method for the determination of cholesterol in milk and other opaque matrices. The initial step of the method is to liberate chemically and physically bound cholesterol from the milk fat globule membrane by enzymatic action. The method is able...... to discriminate between esterified and free cholesterol in milk. The analysis is cost effective and is developed to work directly on whole, fresh milk thereby eliminating time consuming and tedious pre-treatment procedures of the sample. More than 1000 milk samples were analysed on the day of sampling. The total...... concentration of milk cholesterol ranged from 80 to 756 μM (n = 1068; mean 351 μM). Milk cholesterol was significantly correlated to milk fat concentration as analysed by mid-infra red spectrometry (r = 0.630; n = 853) and by an enzymatic–fluorometric method (triacylglycerol) (r = 0.611; n = 842)....

  7. Rapid tryptic mapping using enzymatically active mass spectrometer probe tips

    Energy Technology Data Exchange (ETDEWEB)

    Dogruel, D.; Williams, P.; Nelson, R.W. [Arizona State Univ., Tempe, AZ (United States)

    1995-12-01

    A method has been developed for rapid, sensitive, and accurate tryptic mapping of polypeptides using matrix-assisted laser desorption/ionization time-of-flight mass analysis. The technique utilizes mass spectrometer probe tips which have been activated through the covalent immobilization of trypsin. The enzymatically active probe tips were used for the tryptic mapping of chicken egg lysozyme and the results compared with those obtained using either free trypsin or agarose-immobilized trypsin. A significant increase in the overall sensitivity of the process was observed using the active probe tips, as well as the production of more characteristic proteolytic fragments and the elimination of background signals due to the autolysis of the trypsin. Further, probe tip digestions were found to be rapid and convenient. 19 refs., 6 figs., 2 tabs.

  8. Enzymatic processes in alternative reaction media: a mini review

    Directory of Open Access Journals (Sweden)

    Mansour Ghaffari-Moghaddam

    2015-08-01

    Full Text Available Biocatalysis is a growing field in the production of fine chemicals and will most probably increase its share in the future. Enzymatic reactions are carried out under mild conditions, i.e., non-toxic solvents, low temperature and pressure, which eliminates most environmental drawbacks associated with conventional production methods. The superiority of chemo-, regio- and enantioselectivity of enzymes exhibit significant advantages over conventional catalysts for production of fine chemicals, flavors, fragrances, agrochemicals and pharmaceuticals. Enzymes can function both in aqueous and non-aqueous solvents. As a result of the growing scientific and industrial interest towards green chemistry, green solvent systems, which are mainly water, supercritical fluids, ionic liquids, fluorinated solvents, and solvent-free systems have become more popular in biocatalysis. However, the activity and selectivity of an enzyme is heavily dependent on solvent properties. In this review, various green solvents were classified and some of their influential features on enzyme activity were discussed.

  9. Synthesis of tritium labelled nucleoside triphosphates by enzymatic phosphorylation

    International Nuclear Information System (INIS)

    Shen Decun; Ji Linzhen; Liao Sha

    1986-01-01

    [5- 3 H]UMP, [5- 3 H]CMP, [8- 3 H]AMP and [8- 3 H]GMP were prepared from 5BrUMP 5BrCMP 8BrAMP and 8BrGMP by catalytic halogentritium replacement at the same time. [5- 3 H]UTP, [5- 3 H]CTP, [8- 3 H]ATP and [8- 3 H]GTP were subsequently synthesized from [5- 3 H]UMP, [5- 3 H]CMP, [8- 3 H]AMP and [8- 3 H]GMP by enzymatic phosphorylation with the crude enzyme prepared from brewer's yeasts and purified by paper chromatography simultaneously. In addition, four kinds of tritium labelled nucleoside monophosphates and four kinds of tritium labelled nucleoside diphosphates were obtained as the by-products. The specific activity of these products is between 14-19 Ci/mmol and the radiochemical purity is more than 98%

  10. Production of resistant starch by enzymatic debranching in legume flours.

    Science.gov (United States)

    Morales-Medina, Rocío; Del Mar Muñío, María; Guadix, Emilia M; Guadix, Antonio

    2014-01-30

    Resistant starch (RS) was produced by enzymatic hydrolysis of flours from five different legumes: lentil, chickpea, faba bean, kidney bean and red kidney bean. Each legume was firstly treated thermally, then hydrolyzed with pullulanase for 24h at 50°C and pH 5 and lyophilized. At the end of each hydrolysis reaction, the RS amount ranged from 4.7% for red kidney beans to 7.5% for chickpeas. With respect to the curves of RS against hydrolysis time, a linear increase was observed initially and a plateau was generally achieved by the end of reaction. These curves were successfully modeled by a kinetic equation including three parameters: initial RS, RS at long operation time and a kinetic constant (k). Furthermore, the relative increase in hydrolysis, calculated using the kinetic parameters, was successfully correlated to the percentage of amylose. Copyright © 2013 Elsevier Ltd. All rights reserved.

  11. Enzymatic Modification of Corn Starch Influences Human Fecal Fermentation Profiles.

    Science.gov (United States)

    Dura, Angela; Rose, Devin J; Rosell, Cristina M

    2017-06-14

    Enzymatically modified starches have been widely used in food applications to develop new products, but information regarding digestion and fecal fermentation of these products is sparse. The objective of this study was to determine the fermentation properties of corn starch modified with α-amylase, amyloglucosidase, or cyclodextrin glycosyltransferase and the possible role of hydrolysis products. Samples differed in their digestibility and availability to be fermented by the microbiota, resulting in differences in microbial metabolites produced during in vitro fermentation. The presence or absence of hydrolysis products and gelatinization affected starch composition and subsequent metabolite production by the microbiota. Amyloglucosidase-treated starch led to the greatest production of short- and branched-chain fatty acid production by the microbiota. Results from this study could be taken into consideration to confirm the possible nutritional claims and potential health benefits of these starches as raw ingredients for food development.

  12. Functional bio-based polyesters by enzymatic polymerization

    DEFF Research Database (Denmark)

    Daugaard, Anders Egede; Hoffmann, Christian; Andersen, Christian

    During recent years enzymatic polymerization has become increasingly popular as an alternative to classical polyesterification processes. The high regioselectivity observed for lipases permits preparation of novel polyesters with a high number of functional groups.1 This is particularly interesting...... polymerization was applied to prepare functional water soluble polyesters based on dimethyl itaconate and poly(ethyleneglycol).2 The monomer permits postfunctionalization using thiol-ene chemistry or aza-michael additions, which was used to illustrate the possibilites of preparing functional hydrogels. Hydrogels...... based on the polyesters were shown to be degradable and could be prepared either from the pure polyester or from prefunctionalized polyesters, though the thiol-ene reactions were found to be less effective. Since then a new monomer, trans-2,5-dihydroxy-3-pentenoic acid methyl ester (DPM) has been...

  13. Advances in the enzymatic production of L-hexoses.

    Science.gov (United States)

    Chen, Ziwei; Zhang, Wenli; Zhang, Tao; Jiang, Bo; Mu, Wanmeng

    2016-08-01

    Rare sugars have recently drawn attention because of their potential applications and huge market demands in the food and pharmaceutical industries. All L-hexoses are considered rare sugars, as they rarely occur in nature and are thus very expensive. L-Hexoses are important components of biologically relevant compounds as well as being used as precursors for certain pharmaceutical drugs and thus play an important role in the pharmaceutical industry. Many general strategies have been established for the synthesis of L-hexoses; however, the only one used in the biotechnology industry is the Izumoring strategy. In hexose Izumoring, four entrances link the D- to L-enantiomers, ketose 3-epimerases catalyze the C-3 epimerization of L-ketohexoses, and aldose isomerases catalyze the specific bioconversion of L-ketohexoses and the corresponding L-aldohexoses. In this article, recent studies on the enzymatic production of various L-hexoses are reviewed based on the Izumoring strategy.

  14. DEXTRINIZED SYRUPS OBTAINING THROUGH THE ENZYMATIC HYDROLYSIS OF SORGHUM STARCH

    Directory of Open Access Journals (Sweden)

    Leyanis Rodríguez Rodríguez

    2015-10-01

    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.

  15. Preparation of immobilized growing cells and enzymatic hydrolysis of sawdust

    International Nuclear Information System (INIS)

    Kumakura, M.; Kaetsu, I.

    1984-01-01

    Trichoderma reesei cells were immobilized by radiation polymerization using porous materials such as non-woven material and sawdust, and the enzymatic hydrolysis of sawdust with the enzyme solution from the immobilized growing cells was studied. The filter paper activity, which shows the magnitude of cellulase production in the immobilized cells, was comparable with that in the intact cells. The filter paper activity was affected by addition concentration of monomer and porous materials. The cells in the immobilized cells grew to be adhered on the surface of the fibrous polymers. Sawdust, which was pretreated by irradiation technique, was effectively hydrolyzed with the enzyme solution resulting from the culture of the immobilized cells, in which the glucose yield increased increasing the culture time of the immobilized cells. (author)

  16. Enzymatic Synthesis and Anti-Allergic Activities of Curcumin Oligosaccharides

    Directory of Open Access Journals (Sweden)

    Kei Shimoda

    2010-01-01

    Full Text Available Curcumin 4‘- O -glucooligosaccharides were synthesized by a two step-enzymatic method using almond β-glucosidase and cyclodextrin glucanotransferase (CGTase. Curcumin was glucosylated to curcumin 4‘- O -β-D-glucopyranoside by almond β-glucosidase in 19% yield. Curcumin 4‘- O -β-D-glucopyranoside was converted into curcumin 4‘- O -β-glucooligosaccharides, i.e. 4‘- O -β-maltoside (51% and 4‘- O -β-maltotrioside (25%, by further CGTase-catalyzed glycosylation. Curcumin 4‘- O -β-glycosides showed suppressive action on IgE antibody formation and inhibitory effects on histamine release from rat peritoneal mast cells.

  17. Enzymatic catalysis treatment method of meat industry wastewater using lacasse.

    Science.gov (United States)

    Thirugnanasambandham, K; Sivakumar, V

    2015-01-01

    The process of meat industry produces in a large amount of wastewater that contains high levels of colour and chemical oxygen demand (COD). So they must be pretreated before their discharge into the ecological system. In this paper, enzymatic catalysis (EC) was adopted to treat the meat wastewater. Box-Behnken design (BBD), an experimental design for response surface methodology (RSM), was used to create a set of 29 experimental runs needed for optimizing of the operating conditions. Quadratic regression models with estimated coefficients were developed to describe the colour and COD removals. The experimental results show that EC could effectively reduce colour (95 %) and COD (86 %) at the optimum conditions of enzyme dose of 110 U/L, incubation time of 100 min, pH of 7 and temperature of 40 °C. RSM could be effectively adopted to optimize the operating multifactors in complex EC process.

  18. Beam irradiation pretreatment on enzymatic hydrolysis of biomass

    International Nuclear Information System (INIS)

    Yoo, Hah Young; Choi, Han Suk; Yang, Soo Jeong; Lee, Ja Hyun; Kim, Sung Bong; Jung, Da Un; Kim, Seung Wook

    2013-01-01

    As a renewable energy resource, lignocellulosic biomass has become great attention these days. Miscanthus is considered as one of the best feed stock for sugar production due to its high carbohydrate conversion, more efficient pretreatment process was necessary for removal of enzymatic hydrolysis barriers. In this study, electron beam irradiation pretreatment was utilized to Miscanthus straw for the enhancement of sugar conversion. The prepared samples were exposed 20 ∼ 500 kGy of doses and 5 ∼ 100 kGy of dose rate under 1 MeV of energy. Optimum irradiation conditions were 300 kGy of doses, 10 kGy of doses rate and 7.4 mA of current. Finally, compared with untreated Miscanthus, the glucose conversion was 2 fold increased under optimal conditions

  19. Enzymatic Production of Monoclonal Stoichiometric Single-Stranded DNA Oligonucleotides

    Science.gov (United States)

    Ducani, Cosimo; Kaul, Corinna; Moche, Martin; Shih, William M.; Högberg, Björn

    2013-01-01

    Single-stranded oligonucleotides are important as research tools as probes for diagnostics and gene therapy. Today, production of oligonucleotides is done via solid-phase synthesis. However, the capabilities of current polymer chemistry are limited in comparison to what can be produced in biological systems. The errors in synthetic DNA increases with oligonucleotide length, and sequence diversity can often be a problem. Here, we present the Monoclonal Stoichiometric (MOSIC) method for enzymatic DNA oligonucleotide production. Using this method, we amplify oligonucleotides from clonal templates followed by digestion of a cutter-hairpin, resulting in pools of monoclonal oligonucleotides with precisely controlled relative stoichiometric ratios. We present data where MOSIC oligonucleotides, 14–378 nt long, were prepared either by in vitro rolling-circle amplification, or by amplification in Escherichia coli in the form of phagemid DNA. The formation of a DNA crystal and folding of DNA nanostructures confirmed the scalability, purity and stoichiometry of the produced oligonucleotides. PMID:23727986

  20. Recent Advances in Carbon Nanotube-Based Enzymatic Fuel Cells

    Energy Technology Data Exchange (ETDEWEB)

    Cosnier, Serge, E-mail: serge.cosnier@ujf-grenoble.fr; Holzinger, Michael; Le Goff, Alan [Département de Chimie Moléculaire (DCM) UMR 5250, Université Grenoble Alpes, Grenoble (France); Département de Chimie Moléculaire (DCM) UMR 5250, CNRS, Grenoble (France)

    2014-10-24

    This review summarizes recent trends in the field of enzymatic fuel cells. Thanks to the high specificity of enzymes, biofuel cells can generate electrical energy by oxidation of a targeted fuel (sugars, alcohols, or hydrogen) at the anode and reduction of oxidants (O{sub 2}, H{sub 2}O{sub 2}) at the cathode in complex media. The combination of carbon nanotubes (CNT), enzymes and redox mediators was widely exploited to develop biofuel cells since the electrons involved in the bio-electrocatalytic processes can be efficiently transferred from or to an external circuit. Original approaches to construct electron transfer based CNT-bioelectrodes and impressive biofuel cell performances are reported as well as biomedical applications.

  1. Non-enzymatic glucose detection using magnetic nanoemulsions

    International Nuclear Information System (INIS)

    Mahendran, V.; Philip, John

    2014-01-01

    We probe the optical properties and intermolecular interactions in magnetically responsive nanoemulsions in the presence of glucose. The equilibrium interdroplet distance between the emulsion droplets in an one-dimensional array increases by several nanometers in the presence of glucose because of intermolecular hydrogen bonding with sodium dodecyl sulphate molecules at the oil-water interface that gives rise to stretched lamellae-like structure. The observed large red shift in the diffracted Bragg peak (∼50–100 nm) and the linear response in the glucose concentration range of 0.25–25 mM offer a simple, fast, and cost effective non-enzymatic approach for glucose detection.

  2. Rapid enzymatic response to compensate UV radiation in copepods.

    Directory of Open Access Journals (Sweden)

    María Sol Souza

    Full Text Available Ultraviolet radiation (UVR causes physical damage to DNA, carboxylation of proteins and peroxidation of lipids in copepod crustaceans, ubiquitous and abundant secondary producers in most aquatic ecosystems. Copepod adaptations for long duration exposures include changes in behaviour, changes in pigmentation and ultimately changes in morphology. Adaptations to short-term exposures are little studied. Here we show that short-duration exposure to UVR causes the freshwater calanoid copepod, Eudiaptomus gracilis, to rapidly activate production of enzymes that prevent widespread collateral peroxidation (glutathione S-transferase, GST, that regulate apoptosis cell death (Caspase-3, Casp-3, and that facilitate neurotransmissions (cholinesterase-ChE. None of these enzyme systems is alone sufficient, but they act in concert to reduce the stress level of the organism. The interplay among enzymatic responses provides useful information on how organisms respond to environmental stressors acting on short time scales.

  3. Enzymatic synthesis of C-11 formaldehyde: concise communication

    International Nuclear Information System (INIS)

    Slegers, G.; Lambrecht, R.H.D.; Vandewalle, T.; Meulewaeter, L.; Vandecasteele, C.

    1984-01-01

    An enzymatic synthesis of C-11 formaldehyde from C-11 methanol is presented, with immobilized alcohol oxidase and catalase: a rapid, simple procedure, with a high and reproducible yield. Carbon-11 methanol is oxidized to C-11 formaldehyde by passage over a column on which the enzymes alcohol oxidase and catalase are immobilized. The catalase increases reaction velocity by recycling the oxygen, and prevents destruction of the alcohol oxidase by eliminating the excess of hydrogen peroxide. The yield of the enzyme-catalyzed oxidation was 80-95%. A specific activity of 400-450 mCi/μmole was obtained at EOB + 20 min. Various immobilization techniques and the optimal reaction conditions of the immobilized enzymes are investigated

  4. Oxidative enzymatic gelation of sugar beet pectin for emulsion stabilization

    DEFF Research Database (Denmark)

    Abang Zaidel, Dayang Norulfairuz; Meyer, Anne S.

    2013-01-01

    Pectin from sugar beet is derived from the sugar beet pulp residue which results when sugar beets are processed for sucrose extraction. The sugar beet pectin has poor gelationability by the classic divalentcation molecular mechanism because of a relatively high acetylation degree and short...... polygalacturonate backbone chain length. However, due to the feruloyl-substitutions on the side chains, the sugar beet pectic polysaccharides can be cross-linked via enzyme catalyzed oxidation. The enzyme kinetics and functionality of such oxidativelycross-linked sugar beet pectin, in relation to stabilizing...... emulsions has recently been investigated in model food emulsions. This paper reviews the pectin chemistry, enzymatic oxidative gelation mechanisms, interaction mechanisms of the sugar beet pectin with the emulsion droplets and explores how the gelation affects the rheology and stability of emulsion systems...

  5. Enzymatic gelation of sugar beet pectin in food products

    DEFF Research Database (Denmark)

    Bergsøe, Merete Norsker; Jensen, Mette; Adler-Nissen, Jens

    2000-01-01

    Sugar beet pectin is a food ingredient with specific functional properties. It may form gels by an oxidative cross-linking of ferulic acid. In the present study, the gel forming properties of three oxidative enzymes were examined in different food relevant conditions. The enzymes chosen were two...... laccases and one peroxidase. The textural properties of the produced gels were measured on a texture analyser. The influence of sugar, salt and protein were analysed. Finally, the enzymatic gelation was studied in three food products with added sugar beet pectin. These were black currant juice, milk...... and chopped heat-treated meat emulsion. The addition of salt resulted in softer, less stiff and chewy, and less adhesive gels. Generally speaking, sugar addition increased the hardness but at high concentration the gels were very brittle. However, Young's modulus was lower in gels containing sugar than...

  6. Intramolecular epistasis and the evolution of a new enzymatic function.

    Directory of Open Access Journals (Sweden)

    Sajid Noor

    Full Text Available Atrazine chlorohydrolase (AtzA and its close relative melamine deaminase (TriA differ by just nine amino acid substitutions but have distinct catalytic activities. Together, they offer an informative model system to study the molecular processes that underpin the emergence of new enzymatic function. Here we have constructed the potential evolutionary trajectories between AtzA and TriA, and characterized the catalytic activities and biophysical properties of the intermediates along those trajectories. The order in which the nine amino acid substitutions that separate the enzymes could be introduced to either enzyme, while maintaining significant catalytic activity, was dictated by epistatic interactions, principally between three amino acids within the active site: namely, S331C, N328D and F84L. The mechanistic basis for the epistatic relationships is consistent with a model for the catalytic mechanisms in which protonation is required for hydrolysis of melamine, but not atrazine.

  7. Enzymatic digestibility of peptides cross-linked by ionizing radiation

    International Nuclear Information System (INIS)

    Dizdaroglu, M.; Gajewski, E.; Simic, M.G.

    1984-01-01

    Digestibility by proteolytic enzymes of peptides cross-linked by ionizing radiation was investigated. Small peptides of alanine and phenylalanine were chosen as model compounds and aminopeptidases and carboxypeptidases were used as proteolytic enzymes. Peptides exposed to γ-radiation in aqueous solution were analysed by high-performance liquid chromatography before and after hydrolysis by aminopeptidase M, leucine aminopeptidase carboxypeptidase A and carboxypeptidase Y. The results obtained clearly demonstrate the different actions of these enzymes on cross-linked aliphatic and aromatic peptides. Peptide bonds of cross-linked dipeptides of alanine were completely resistant to enzymatic hydrolysis whereas the enzymes, except for carboxypeptidase Y, cleaved all peptide bonds of cross-linked peptides of phenylalanine. The actions of the enzymes on these particular compounds are discussed in detail. (author)

  8. Enzymatic transesterification of waste vegetable oil to produce biodiesel.

    Science.gov (United States)

    Lopresto, C G; Naccarato, S; Albo, L; De Paola, M G; Chakraborty, S; Curcio, S; Calabrò, V

    2015-11-01

    An experimental study on enzymatic transesterification was performed to produce biodiesel from waste vegetable oils. Lipase from Pseudomonas cepacia was covalently immobilized on a epoxy-acrylic resin support. The immobilized enzyme exhibited high catalytic specific surface and allowed an easy recovery, regeneration and reutilisation of biocatalyst. Waste vegetable oils - such as frying oils, considered not competitive with food applications and wastes to be treated - were used as a source of glycerides. Ethanol was used as a short chain alcohol and was added in three steps with the aim to reduce its inhibitory effect on lipase activity. The effect of biocatalyst/substrate feed mass ratios and the waste oil quality have been investigated in order to estimate the process performances. Biocatalyst recovery and reuse have been also studied with the aim to verify the stability of the biocatalyst for its application in industrial scale. Copyright © 2015 Elsevier Inc. All rights reserved.

  9. Transport equations in an enzymatic glucose fuel cell

    Science.gov (United States)

    Jariwala, Soham; Krishnamurthy, Balaji

    2018-01-01

    A mathematical model is developed to study the effects of convective flux and operating temperature on the performance of an enzymatic glucose fuel cell with a membrane. The model assumes isothermal operating conditions and constant feed rate of glucose. The glucose fuel cell domain is divided into five sections, with governing equations describing transport characteristics in each region, namely - anode diffusion layer, anode catalyst layer (enzyme layer), membrane, cathode catalyst layer and cathode diffusion layer. The mass transport is assumed to be one-dimensional and the governing equations are solved numerically. The effects flow rate of glucose feed on the performance of the fuel cell are studied as it contributes significantly to the convective flux. The effects of operating temperature on the performance of a glucose fuel cell are also modeled. The cell performances are compared using cell polarization curves, which were found compliant with experimental observations.

  10. Effect of Maize Biomass Composition on the Optimization of Dilute-Acid Pretreatments and Enzymatic Saccharification

    NARCIS (Netherlands)

    Torres Salvador, A.F.; Weijde, van der R.T.; Dolstra, O.; Visser, R.G.F.; Trindade, L.M.

    2013-01-01

    At the core of cellulosic ethanol research are innovations leading to reductions in the chemical and energetic stringency of thermochemical pretreatments and enzymatic saccharification. In this study, key compositional features of maize cell walls influencing the enzymatic conversion of biomass into

  11. 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

    2011-01-01

    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 ...... the initial part of enzymatic hydrolysis of cellulose. The implications of this phenomenon have not yet been recognized or explored within cellulosic biofuels....

  12. Xylanase supplementation on enzymatic saccharification of dilute acid pretreated poplars at different severities

    Science.gov (United States)

    Chao Zhang; Xinshu Zhuang; Zhao Jiang Wang; Fred Matt; Franz St. John; J.Y. Zhu

    2013-01-01

    Three pairs of solid substrates from dilute acid pretreatment of two poplar wood samples were enzymatically hydrolyzed by cellulase preparations supplemented with xylanase. Supplementation of xylanase improved cellulose saccharification perhaps due to improved cellulose accessibility by xylan hydrolysis. Total xylan removal directly affected enzymatic cellulose...

  13. Rapid and sensitive enzymatic-radiochemical assay for the determination of triglycerides

    International Nuclear Information System (INIS)

    Khoo, J.C.; Miller, E.; Goldberg, D.I.

    1987-01-01

    An enzymatic-radiochemical method suitable for the determination of triglyceride levels of cells in culture is described. The method is based on the enzymatic hydrolysis of triglycerides to free fatty acids which then complex with 63 Ni. The method is rapid, accurate, and inexpensive. The procedure extends the sensitivity of triglyceride measurement to as low as 0.25 nanomoles

  14. Enzymatic pH Control for Biomimetic Deposition of Calcium Phosphate Coatings

    NARCIS (Netherlands)

    Nijhuis, A.W.; Reza Nejadnik, M.; Nudelman, F.; Walboomers, X.F.; te Riet, J.; Habibovic, Pamela; Tahmasebi Birgani, Zeinab; Yubao, L.; Bomans, P.H.H.; Jansen, J.A.; Sommerdijk, N.A.J.M.; Leeuwenburgh, S.C.G.

    2014-01-01

    The current study has focused on enzymatic decomposition of urea into carbon dioxide and ammonia as a means to increase the pH during biomimetic deposition of Calcium Phospate (CaP) onto implant surfaces. The kinetics of the enzymatically induced pH increase were studied by monitoring pH, calcium

  15. Enzymatic pH control for biomimetic depostion of calcium phosphate coatings

    NARCIS (Netherlands)

    Nijhuis, A.W.G.; Nejadnik, M.R.; Nudelman, F.; Walboomers, X.F.; Riet, te J.; Habibovic, P.; Birgani, Z.T.; Li, Y.B.; Bomans, P.H.H.; Jansen, J.A.; Sommerdijk, N.A.J.M.; Leeuwenburgh, S.C.G.

    2014-01-01

    The current study examines the enzymatic decomposition of urea into carbon dioxide and ammonia as a means to increase the pH during biomimetic deposition of calcium phospate (CaP) onto implant surfaces. The kinetics of the enzymatically induced pH increase were studied by monitoring pH, calcium

  16. Enzymatic pH control for biomimetic deposition of calcium phosphate coatings

    NARCIS (Netherlands)

    Nijhuis, A.W.G.; Nejadnik, M.R.; Nudelman, F.; Walboomers, X.F.; Riet, J. te; Habibovic, P.; Tahmasebi Birgani, Z.; Li, Y.; Bomans, P.H.; Jansen, J.A.; Sommerdijk, N.A.; Leeuwenburgh, S.C.G.

    2014-01-01

    The current study examines the enzymatic decomposition of urea into carbon dioxide and ammonia as a means to increase the pH during biomimetic deposition of calcium phosphate (CaP) onto implant surfaces. The kinetics of the enzymatically induced pH increase were studied by monitoring pH, calcium

  17. The effect of high intensity mixing on the enzymatic hydrolysis of concentrated cellulose fiber suspensions

    Science.gov (United States)

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

    2011-01-01

    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...

  18. Understanding the effects of lignosulfonate on enzymatic saccharification of pure cellulose

    Science.gov (United States)

    Hongming Lou; Haifeng Zhou; Xiuli Li; Mengxia Wang; J.Y. Zhu; Xueqing Qiu

    2014-01-01

    The effects of lignosulfonate (LS) on enzymatic saccharification of pure cellulose were studied. Four fractions of LS with different molecular weight (MW) prepared by ultrafiltration of a commercial LS were applied at different loadings to enzymatic hydrolysis of Whatman paper under different pH. Using LS fractions with low MW and high degree of sulfonation can enhance...

  19. Cost analysis of enzymatic biodiesel production in small-scaled packed-bed reactors

    NARCIS (Netherlands)

    Budzaki, S.; Miljic, G.; Sundaram, S.; Tisma, M.; Hessel, V.

    2017-01-01

    A cost analysis of enzymatic biodiesel production in small-scaled packed-bed reactors using refined sunflower oil is performed in this work. A few enzymatic micro-flow reactors have so far reached a performance close to gram-scale, which might be sufficient for the pharmaceutical industry. This

  20. Enzymatic collection test as total gamma irradiation pronostic test in rat, rabbit and man

    International Nuclear Information System (INIS)

    Breuil, G.; Dinnequin, B.

    The purpose of this study is to known, during 30 days, what becomes the animal whose enzymatic co-ordinates are well known. Both 100, 160, 200, 325, 400, 650, 850, 975, 1000, 1300 rads irradiated rabbit serum enzymatic evolution and that of two 1000 rads in toto irradiated leucemic men for a cord graft are studied [fr

  1. Chemical catalysis in biodiesel production (I): enzymatic catalysis processes

    International Nuclear Information System (INIS)

    Jachmarian, I.; Dobroyan, M.; Veira, J.; Vieitez, I.; Mottini, M.; Segura, N.; Grompone, M.

    2009-01-01

    There are some well known advantages related with the substitution of chemical catalysis by enzymatic catalysis processes.Some commercial immobilized lipases are useful for the catalysis of bio diesel reaction, which permits the achievement of high conversions and the recovery of high purity products, like a high quality glycerine. The main disadvantage of this alternative method is related with the last inactivation of the enzyme (by both the effect of the alcohol and the absorption of glycerol on catalyst surface), which added to the high cost of the catalyst, produces an unfavourable economical balance of the entire process. In the work the efficiency of two commercial immobilized lipases (Lipozyme TL IM y Novozyme 435 NNovozymes-Dinamarca) in the catalysis of the continuous transesterification of sunflower oil with different alcohols was studied. The intersolubility of the different mixturesinvolving reactans (S oil/alkyl esters/alcohol) and products (P mixtures with a higher content of 1% of glycerol,while for ethanol homogeneous mixtures were obtained at 12% of glycerol (44.44 12).Using and ethanolic substrate at the proportion S=19:75:6 and Lipozyme TL IM, it was possible to achieve a 98% of convertion to the corresponding biodiesel.When Novozymes 435 catalyzed the process it was possible to increase the oil concentration in the substrateaccording to proportion S=35:30:35, and a 78% conversion was obtained. The productivity shown by the firt enzyme was 70mg biodiesel g enzime-1, hora-1 while with the second one the productivity increased to 230. Results suggested that the convenient adjustement of substrate composition with the addition of biodiesel to reactants offers an efficient method for maximizing the enzyme productivity, hence improving the profitability of the enzymatic catalyzed process. (author)

  2. Optimal information transfer in enzymatic networks: A field theoretic formulation

    Science.gov (United States)

    Samanta, Himadri S.; Hinczewski, Michael; Thirumalai, D.

    2017-07-01

    Signaling in enzymatic networks is typically triggered by environmental fluctuations, resulting in a series of stochastic chemical reactions, leading to corruption of the signal by noise. For example, information flow is initiated by binding of extracellular ligands to receptors, which is transmitted through a cascade involving kinase-phosphatase stochastic chemical reactions. For a class of such networks, we develop a general field-theoretic approach to calculate the error in signal transmission as a function of an appropriate control variable. Application of the theory to a simple push-pull network, a module in the kinase-phosphatase cascade, recovers the exact results for error in signal transmission previously obtained using umbral calculus [Hinczewski and Thirumalai, Phys. Rev. X 4, 041017 (2014), 10.1103/PhysRevX.4.041017]. We illustrate the generality of the theory by studying the minimal errors in noise reduction in a reaction cascade with two connected push-pull modules. Such a cascade behaves as an effective three-species network with a pseudointermediate. In this case, optimal information transfer, resulting in the smallest square of the error between the input and output, occurs with a time delay, which is given by the inverse of the decay rate of the pseudointermediate. Surprisingly, in these examples the minimum error computed using simulations that take nonlinearities and discrete nature of molecules into account coincides with the predictions of a linear theory. In contrast, there are substantial deviations between simulations and predictions of the linear theory in error in signal propagation in an enzymatic push-pull network for a certain range of parameters. Inclusion of second-order perturbative corrections shows that differences between simulations and theoretical predictions are minimized. Our study establishes that a field theoretic formulation of stochastic biological signaling offers a systematic way to understand error propagation in

  3. Hydrolytic And Enzymatic Degradation Characteristics Of Biodegradable Aliphatic Polysters

    Institute of Scientific and Technical Information of China (English)

    LI Suming

    2004-01-01

    Aliphatic polyesters, especially those derived from lactide (PLA), glycolide (PGA) and ε-caprolactone (PCL), are being investigated worldwide for applications in the field of surgery (suture material, devices for internal bone fracture fixation), pharmacology (sustained drug delivery systems), and tissue engineering (scaffold for tissue regeneration) [1,2]. This is mainly due to their good biocompatibility and variable degradability. These polymers present also a growing interest for environmental applications in agriculture (mulch films) and in our everyday life (packaging material)as the development of biodegradable materials is now considered as one of the potential solutions to the problem of plastic waste management.For both biomedical and environmental applications, it is of major importance to understand the degradation characteristics of the polymers. The hydrolytic degradation of aliphatic polyesters has been investigated by many research groups. Our group has shown that degradation of PLAGA large size devices is faster inside than at the surface. This heterogeneous degradation is due to the autocatalytic effect of carboxylic endgroups formed by ester bond cleavage. Moreover,degradation-induced morphological and compositional changes were also elucidated. In the case of PCL, the hydrolytic degradation is very slow due to its hydrophobicity and crystallinity.The enzymatic degradation of these polymers has been investigated by a number of authors. A specific enzyme, proteinase K, has been shown to have significant effects on PLA degradation. This enzyme preferentially degrade L-lactate units as opposed to D-lactate ones, amorphous zones as opposed to crystalline ones [3]. The enzymatic degradation of PCL polymers has also been investigated. A number of lipase-type enzymes were found to significantly accelerate the degradation of PCL despite its high crystallinity. In the case of PLA/PCL blends, the two components exhibited well separated crystalline domains

  4. Study of Enzymatic Hydrolysis of Dilute Acid Pretreated Coconut Husk

    Directory of Open Access Journals (Sweden)

    Rudy Agustriyanto

    2012-12-01

    Full Text Available Coconut husk is classified as complex lignocellulosic material that contains cellulose, hemicellulose, lignin, and some other extractive compounds. Cellulose from coconut husk can be used as fermentation substrate after enzymatic hydrolysis. In contrary, lignin content from the coconut husk will act as an inhibitor in this hydrolysis process. Therefore, a pretreatment process is needed to enhance the hydrolysis of cellulose. The objective of this research is to investigate the production of the glucose through dilute acid pretreatment and to obtain its optimum operating conditions. In this study, the pretreatment was done using dilute sulfuric acid in an autoclave reactor. The pretreatment condition were varied at 80°C, 100°C, 120°C and 0.9%, 1.2%, 1.5% for temperature and acid concentration respectively. The acid pretreated coconut husk was then hydrolyzed using commercial cellulase (celluclast and β-glucosidase (Novozyme 188. The hydrolysis time was 72 hours and the operating conditions were varied at several temperature and pH. From the experimental results it can be concluded that the delignification temperature variation has greater influence than the acid concentration. The optimum operating condition was obtained at pH 4 and 50°C which was pretreated at 100°C using 1.5% acid concentration. Copyright © 2012 by BCREC UNDIP. All rights reserved. (Selected Paper from International Conference on Chemical and Material Engineering (ICCME 2012Received: 28th September 2012, Revised: 2nd October 2012, Accepted: 4th October 2012[How to Cite: R. Agustriyanto, A. Fatmawati, Y. Liasari. (2012. Study of Enzymatic Hydrolysis of Dilute Acid Pretreated Coconut Husk. Bulletin of Chemical Reaction Engineering & Catalysis, 7(2: 137-141. doi:10.9767/bcrec.7.2.4046.137-141] [How to Link / DOI: http://dx.doi.org/10.9767/bcrec.7.2.4046.137-141 ] | View in 

  5. Acute hormonal, immunological and enzymatic responses to a basketball game

    Directory of Open Access Journals (Sweden)

    Denis Foschini

    2008-01-01

    Full Text Available The objective of the present study was to analyze the acute hormonal, immunological and enzymatic responses of professional basketball players to a basketball game. The sample was composed of eight basketball athletes, with a minimum of 4 years’ experience in basketball. A real game was simulated with a total duration of 40 minutes, divided into two halves of 20 minutes each and an interval of 10 minutes between halves. Blood samples were collected before and immediately after the game (20 ml, vacuum tube system. The variables analyzed were: testosterone and cortisol hormones, total leukocytes, neutrophils, lymphocytes, monocytes and the enzymes creatine kinase (CK and lactate dehydrogenase (LDH. Statistical analysis was with descriptive statistics and the Student’s t test for paired samples to p≤0.05. The pre (13.34 nmol/L and 301.97 nmol/L and post game (17.34 nmol/L and 395.91 nmol/L levels of testosterone and cortisol were statistically different, with higher levels after the game for both hormones. The immune cell counts exhibited significant differences for total leukocytes (6393.75 nmol/L and 9158.75 nmol/L and neutrophils (3532.5 nmol/L and 6392.62 nmol/L, with levels being higher after the game. No statistical differences were observed for the enzymatic variables. Therefore, based on the markers analyzed, testosterone and cortisol exhibited pronounced increases after the game and the same behavior was observed for total leukocytes and neutrophils.

  6. Optimization of the Enzymatic Saccharification Process of Milled Orange Wastes

    Directory of Open Access Journals (Sweden)

    Daniel Velasco

    2017-08-01

    Full Text Available Orange juice production generates a very high quantity of residues (Orange Peel Waste or OPW-50–60% of total weight that can be used for cattle feed as well as feedstock for the extraction or production of essential oils, pectin and nutraceutics and several monosaccharides by saccharification, inversion and enzyme-aided extraction. As in all solid wastes, simple pretreatments can enhance these processes. In this study, hydrothermal pretreatments and knife milling have been analyzed with enzyme saccharification at different dry solid contents as the selection test: simple knife milling seemed more appropriate, as no added pretreatment resulted in better final glucose yields. A Taguchi optimization study on dry solid to liquid content and the composition of the enzymatic cocktail was undertaken. The amounts of enzymatic preparations were set to reduce their impact on the economy of the process; however, as expected, the highest amounts resulted in the best yields to glucose and other monomers. Interestingly, the highest content in solid to liquid (11.5% on dry basis rendered the best yields. Additionally, in search for process economy with high yields, operational conditions were set: medium amounts of hemicellulases, polygalacturonases and β-glucosidases. Finally, a fractal kinetic modelling of results for all products from the saccharification process indicated very high activities resulting in the liberation of glucose, fructose and xylose, and very low activities to arabinose and galactose. High activity on pectin was also observed, but, for all monomers liberated initially at a fast rate, high hindrances appeared during the saccharification process.

  7. Acute hormonal, immunological and enzymatic responses to a basketball game

    Directory of Open Access Journals (Sweden)

    Denis Foschini

    2008-12-01

    Full Text Available The objective of the present study was to analyze the acute hormonal, immunological and enzymatic responses of professional basketball players to a basketball game. The sample was composed of eight basketball athletes, with a minimum of 4 years’ experience in basketball. A real game was simulated with a total duration of 40 minutes, divided into two halves of 20 minutes each and an interval of 10 minutes between halves. Blood samples were collected before andimmediately after the game (20 ml, vacuum tube system. The variables analyzed were: testosterone and cortisol hormones, total leukocytes, neutrophils, lymphocytes, monocytes and the enzymes creatine kinase (CK and lactate dehydrogenase (LDH. Statistical analysis was with descriptive statistics and the Student’s t test for paired samples to p≤0.05. The pre (13.34 nmol/L and 301.97 nmol/L and post game (17.34 nmol/L and 395.91 nmol/L levels of testosterone and cortisol were statistically different, with higher levels after the game for both hormones. The immune cell counts exhibited significant differences for total leukocytes (6393.75 nmol/L and 9158.75 nmol/L and neutrophils (3532.5 nmol/L and 6392.62 nmol/L, with levels being higher after the game. No statistical differences were observed for the enzymatic variables. Therefore, based on the markers analyzed, testosterone and cortisol exhibited pronounced increases after the game and the samebehavior was observed for total leukocytes and neutrophils.

  8. [Non-enzymatic glycosylation of dietary protein in vitro].

    Science.gov (United States)

    Bednykh, B S; Evdokimov, I A; Sokolov, A I

    2015-01-01

    Non-enzymatic glycosylation of proteins, based on discovered by Mayarn reaction of carbohydrate aldehyde group with a free amino group of a protein molecule, is well known to experts in biochemistry of food industry. Generated brown solid in some cases give the product marketable qualities--crackling bread--in others conversely, worsen the product. The biological effects of far-advanced products of non-enzymatic protein glycosylation reaction have not been studied enough, although it was reported previously that they are not split by digestive enzymes and couldn't be absorbed by animals. The objective of this work was to compare the depth of glycosylation of different food proteins of animal and vegetable origin. The objects of the study were proteins of animal (casein, lactoglobulin, albumin) and vegetable (soy isolate, proteins of rice flour, buckwheat, oatmeal) origin, glucose and fructose were selected as glycosylation agents, exposure 15 days at 37 degrees C. Lactoglobulin was glycosylated to a lesser extent among the proteins of animal origin while protein of oatmeal was glycosylated in the least degree among vegetable proteins. Conversely, such proteins as casein and soya isolate protein bound rather large amounts of carbohydrates. Fructose binding with protein was generally higher than the binding of glucose. The only exception was a protein of oatmeal. When of glucose and fructose simultaneously presented in the incubation medium, glucose binding usually increased while binding of fructose, in contrast, reduced. According to the total amount of carbohydrate (mcg), which is able to attach a protein (mg) the studied food proteins located in the following order: albumin (38) > soy protein isolate (23) > casein (15,) > whey protein rice flour protein (6) > protein from buckwheat flour (3) > globulin (2) > protein of oatmeal (0.3). The results obtained are to be used to select the optimal combination of proteins and carbohydrates, in which the glycosylation

  9. Identification of Key Residues for Enzymatic Carboxylate Reduction

    Directory of Open Access Journals (Sweden)

    Holly Stolterfoht

    2018-02-01

    Full Text Available Carboxylate reductases (CARs, E.C. 1.2.1.30 generate aldehydes from their corresponding carboxylic acid with high selectivity. Little is known about the structure of CARs and their catalytically important amino acid residues. The identification of key residues for carboxylate reduction provides a starting point to gain deeper understanding of enzymatic carboxylate reduction. A multiple sequence alignment of CARs with confirmed activity recently identified in our lab and from the literature revealed a fingerprint of conserved amino acids. We studied the function of conserved residues by multiple sequence alignments and mutational replacements of these residues. In this study, single-site alanine variants of Neurospora crassa CAR were investigated to determine the contribution of conserved residues to the function, expressability or stability of the enzyme. The effect of amino acid replacements was investigated by analyzing enzymatic activity of the variants in vivo and in vitro. Supported by molecular modeling, we interpreted that five of these residues are essential for catalytic activity, or substrate and co-substrate binding. We identified amino acid residues having significant impact on CAR activity. Replacement of His 237, Glu 433, Ser 595, Tyr 844, and Lys 848 by Ala abolish CAR activity, indicating their key role in acid reduction. These results may assist in the functional annotation of CAR coding genes in genomic databases. While some other conserved residues decreased activity or had no significant impact, four residues increased the specific activity of NcCAR variants when replaced by alanine. Finally, we showed that NcCAR wild-type and mutants efficiently reduce aliphatic acids.

  10. Non-enzymatic palladium recovery on microbial and synthetic surfaces.

    Science.gov (United States)

    Rotaru, Amelia-Elena; Jiang, Wei; Finster, Kai; Skrydstrup, Troels; Meyer, Rikke Louise

    2012-08-01

    The use of microorganisms as support for reduction of dissolved Pd(II) to immobilized Pd(0) nanoparticles is an environmentally friendly approach for Pd recovery from waste. To better understand and engineer Pd(0) nanoparticle synthesis, one has to consider the mechanisms by which Pd(II) is reduced on microbial surfaces. Escherichia coli, Shewanella oneidensis, and Pseudomonas putida were used as model organisms in order to elucidate the role of microbial cells in Pd(II) reduction under acidic conditions. Pd(II) was reduced by formate under acidic conditions, and the process occurred substantially faster in the presence of cells as compared to cell-free controls. We found no difference between native (untreated) and autoclaved cells, and could demonstrate that even a non-enzymatic protein (bovine serum albumin) stimulated Pd(II) reduction as efficiently as bacterial cells. Amine groups readily interact with Pd(II), and to specifically test their role in surface-assisted Pd(II) reduction by formate, we replaced bacterial cells with polystyrene microparticles functionalized with amine or carboxyl groups. Amine-functionalized microparticles had the same effect on Pd(II) reduction as bacterial cells, and the effect could be hampered if the amine groups were blocked by acetylation. The interaction with amine groups was confirmed by infrared spectroscopy on whole cells and amine-functionalized microparticles. In conclusion, bio-supported Pd(II) reduction on microbial surfaces is possibly mediated by a non-enzymatic mechanism. We therefore suggest the use of amine-rich biomaterials rather than intact cells for Pd bio-recovery from waste. Copyright © 2012 Wiley Periodicals, Inc.

  11. pH catalyzed pretreatment of corn bran for enhanced enzymatic arabinoxylan degradation

    DEFF Research Database (Denmark)

    Agger, Jane; Johansen, Katja Salomon; Meyer, Anne S.

    2011-01-01

    Corn bran is mainly made up of the pericarp of corn kernels and is a byproduct stream resulting from the wet milling step in corn starch processing. Through statistic modeling this study examined the optimization of pretreatment of corn bran for enzymatic hydrolysis. A low pH pretreatment (pH 2......, 150°C, 65min) boosted the enzymatic release of xylose and glucose and maximized biomass solubilization. With more acidic pretreatment followed by enzymatic hydrolysis the total xylose release was maximized (at pH 1.3) reaching ∼50% by weight of the original amount present in destarched corn bran......, but the enzyme catalyzed xylose release was maximal after pretreatment at approx. pH 2. The total glucose release peaked after pretreatment of approx. pH 1.5 with an enzymatic release of approx. 68% by weight of the original amounts present in destarched corn bran. For arabinose the enzymatic release...

  12. Structural changes in lignin during organosolv pretreatment of Liriodendron tulipifera and the effect on enzymatic hydrolysis

    International Nuclear Information System (INIS)

    Koo, Bon-Wook; Min, Byeong-Cheol; Gwak, Ki-Seob; Lee, Soo-Min; Choi, Joon-Weon; Yeo, Hwanmyeong; Choi, In-Gyu

    2012-01-01

    Although organosolv pretreatment removed substantial amounts of lignin and xylan, the yield of glucan which is a major sugar source for fermentation to ethanol is more than 90% in most conditions of the organosolv pretreatment. Relative lignin contents of all pretreated biomass were more than 200 g kg −1 , however enzymatic conversions were increased dramatically comparing to untreated biomass. Therefore the correlation between lignin and enzymatic hydrolysis could not be explained just by lignin content, and other changes resulting from lignin removal affected enzymatic hydrolysis. Results on enzymatic conversion and sugar recovery suggested that the critical temperature improving enzymatic hydrolysis significantly was between 120 °C and 130 °C. Microscopic analysis using Field emission scanning electron microscopy (FE-SEM) showed that structural lignin changes happened through organosolv pretreatment. Lignins were isolated from lignin carbohydrate complex (LCC) at the initial stage and then migrated to the surface of biomass. The isolated and migrated lignins were finally redistributed onto surface. These structural changes formed droplets on surface and increased pore volume in pretreated biomass. The increase in pore volume also increased available surface area and enzyme adsorption at initial stage, and thus enzymatic conversion increased significantly through organosolv pretreatment. It was verified that the droplets were mainly composed of lignin and the lignin droplets inhibited enzymatic hydrolysis through adsorption with cellulase. -- Highlights: ► Just lignin contents cannot explain a correlation with enzymatic hydrolysis. ► Several changes resulted from lignin removal must affect enzymatic hydrolysis. ► Droplets are formed by structural changes in lignin during organosolv pretreatment. ► Formation of the lignin droplet increases the pore volume in biomass. ► The increase in pore volume enhances the enzymatic hydrolysis.

  13. Topical formulations with superoxide dismutase: influence of formulation composition on physical stability and enzymatic activity.

    Science.gov (United States)

    Di Mambro, Valéria M; Borin, Maria F; Fonseca, Maria J V

    2003-04-24

    Three different topical formulations were supplemented with superoxide dismutase (SOD) and evaluated concerning physical and chemical stabilities in order to determine the most stable formulation that would maintain SOD activity. Physical stability was evaluated by storing the formulation at room temperature, and at 37 and 45 degrees C for 28 days. Samples were collected at 7-day intervals for assessment of rheological behavior. Chemical stability was evaluated by the measurement of enzymatic activity in formulations stored at room temperature and at 45 degrees C for 75 days. The formulations showed a pseudoplastic behavior, with a flow index of less than 1. There was no significant difference in the initial values of flow index, hysteresis loop or minimum apparent viscosity. The simple emulsion and the one stabilized with hydroxyethylcellulose showed decreased viscosity by the 21st day and with higher temperature, but no significant changes concerning the presence of SOD. Although there were no significant changes concerning storage time or temperature, the formulation stabilized with hydroxyethylcellulose showed a marked loss of SOD activity. The addition of SOD to the formulations studied did not affect their physical stability. Simple emulsions or emulsions stabilized with carboxypolymethylene seem to be better bases for enzyme addition than emulsion stabilized with hydroxyethylcellulose.

  14. An integrated chemo-enzymatic route for preparation of ß-thymidine, a key intermediate in the preparation of antiretrovirals

    CSIR Research Space (South Africa)

    Gordon, GER

    2011-01-01

    Full Text Available A chemo-enzymatic method for production of ß-thymidine, an intermediate in the synthesis of antiretrovirals, is described. Guanosine and thymine were converted by means of enzymatic transglycosylation to yield 5-methyluridine (5-MU), which...

  15. Physiological and enzymatic analyses of pineapple subjected to ionizing radiation; Analises fisiologicas e enzimaticas em abacaxi submetido a tecnologia de radiacao ionizante

    Energy Technology Data Exchange (ETDEWEB)

    Silva, Josenilda Maria da [Centro de Energia Nuclear na Agricultura (CENA), Piracicaba, SP (Brazil)]. E-mail: jmnilda@yahoo.com.br; Silva, Juliana Pizarro; Spoto, Marta Helena Fillet [Escola Superior de Agricultura Luiz de Queiroz (ESALQ/USP), Piracicaba, SP (Brazil)

    2007-07-15

    The physiological and enzymatic post-harvest characteristics of the pineapple cultivar Smooth Cayenne were evaluated after the fruits were gamma-irradiated with doses of 100 and 150 Gy and the fruits were stored for 10, 20 and 30 days at 12 deg C ({+-}1) and relative humidity of 85% ({+-}5). Physiological and enzymatic analyses were made for each storage period to evaluate the alterations resulting from the application of ionizing radiation. Control specimens showed higher values of soluble pectins, total pectins, reducing sugars, sucrose and total sugars and lower values of polyphenyloxidase and polygalacturonase enzyme activities. All the analyses indicated that storage time is a significantly influencing factor. The 100 Gy dosage and 20-day storage period presented the best results from the standpoint of maturation and conservation of the fruits quality. (author)

  16. Improving the enzymatic hydrolysis of thermo-mechanical fiber from Eucalyptus urophylla by a combination of hydrothermal pretreatment and alkali fractionation.

    Science.gov (United States)

    Sun, Shaoni; Cao, Xuefei; Sun, Shaolong; Xu, Feng; Song, Xianliang; Sun, Run-Cang; Jones, Gwynn Lloyd

    2014-01-01

    The recalcitrance of lignocellulosic biomass is a major limitation for its conversion into biofuels by enzymatic hydrolysis. The use of a pretreatment technology is an essential step to diminish biomass recalcitrance for bioethanol production. In this study, a two-step pretreatment using hydrothermal pretreatment at various temperatures and alkali fractionation was performed on eucalyptus fiber. The detailed chemical composition, physicochemical characteristics, and morphology of the pretreated fibers in each of the fractions were evaluated to advance the performance of eucalyptus fiber in enzymatic digestibility. The hydrothermal pretreatment (100 to 220°C) significantly degraded hemicelluloses, resulting in an increased crystallinity of the pretreated fibers. However, as the pretreatment temperature reached 240°C, partial cellulose was degraded, resulting in a reduced crystallinity of cellulose. As compared to the hydrothermal pretreatment alone, a combination of hydrothermal and alkali treatments significantly removed hemicelluloses and lignin, resulting in an improved enzymatic hydrolysis of the cellulose-rich fractions. As compared with the raw fiber, the enzymatic hydrolysis rate increased 1.1 to 8.5 times as the hydrothermal pretreatment temperature increased from 100 to 240°C. Interestingly, after a combination of hydrothermal pretreatment and alkali fractionation, the enzymatic hydrolysis rate increased 3.7 to 9.2 times. Taking into consideration the consumption of energy and the production of xylo-oligosaccharides and lignin, an optimum pretreatment condition was found to be hydrothermal pretreatment at 180°C for 30 min and alkali fractionation with 2% NaOH at 90°C for 2.5 h, in which 66.3% cellulose was converted into glucose by enzymatic hydrolysis. The combination of hydrothermal pretreatment and alkali fractionation was a promising method to remove hemicelluloses and lignin as well as overcome the biomass recalcitrance for enzymatic hydrolysis

  17. Oxidative stability during storage of structured lipids produced from fish oil and caprylic acid

    DEFF Research Database (Denmark)

    Nielsen, Nina Skall; Xu, Xuebing; Timm Heinrich, Maike

    2004-01-01

    Structured lipids produced by enzymatic or chemical methods for different applications have been receiving considerable attention. The oxidative stability of a randomized structured lipid (RFO), produced by chemical interesterification from fish oil (FO) and tricaprylin, and a specific structured...... lipid (SFO), produced by enzymatic interesterification from the same oil and caprylic acid, was compared with the stability of FO. Oils were stored at 2degreesC for 11 wk followed by storage at 20degreesC for 6 wk. In addition, the antioxidative effect of adding the metal chelators EDTA or citric acid...

  18. Testing the applicability of rapid on-site enzymatic activity detection for surface water monitoring

    Science.gov (United States)

    Stadler, Philipp; Vogl, Wolfgang; Juri, Koschelnik; Markus, Epp; Maximilian, Lackner; Markus, Oismüller; Monika, Kumpan; Peter, Strauss; Regina, Sommer; Gabriela, Ryzinska-Paier; Farnleitner Andreas, H.; Matthias, Zessner

    2015-04-01

    On-site detection of enzymatic activities has been suggested as a rapid surrogate for microbiological pollution monitoring of water resources (e.g. using glucuronidases, galactosidases, esterases). Due to the possible short measuring intervals enzymatic methods have high potential as near-real time water quality monitoring tools. This presentation describes results from a long termed field test. For twelve months, two ColiMinder devices (Vienna Water Monitoring, Austria) for on-site determination of enzymatic activity were tested for stream water monitoring at the experimental catchment HOAL (Hydrological Open Air Laboratory, Center for Water Resource Systems, Vienna University of Technology). The devices were overall able to follow and reflect the diverse hydrological and microbiological conditions of the monitored stream during the test period. Continuous data in high temporal resolution captured the course of enzymatic activity in stream water during diverse rainfall events. The method also proofed sensitive enough to determine diurnal fluctuations of enzymatic activity in stream water during dry periods. The method was able to capture a seasonal trend of enzymatic activity in stream water that matches the results gained from Colilert18 analysis for E. coli and coliform bacteria of monthly grab samples. Furthermore the comparison of ColiMinder data with measurements gained at the same test site with devices using the same method but having different construction design (BACTcontrol, microLAN) showed consistent measuring results. Comparative analysis showed significant differences between measured enzymatic activity (modified fishman units and pmol/min/100ml) and cultivation based analyses (most probable number, colony forming unit). Methods of enzymatic activity measures are capable to detect ideally the enzymatic activity caused by all active target bacteria members, including VBNC (viable but nonculturable) while cultivation based methods cannot detect VBNC

  19. Physicochemical and sensory characterization of refined and deodorized tuna (Thunnus albacares) by-product oil obtained by enzymatic hydrolysis.

    Science.gov (United States)

    de Oliveira, Dayse A S B; Minozzo, Marcelo G; Licodiedoff, Silvana; Waszczynskyj, Nina

    2016-09-15

    In this study, the effects of chemical refining and deodorization on fatty acid profiles and physicochemical and sensory characteristics of the tuna by-product oil obtained by enzymatic hydrolysis were evaluated. Enzymatic extraction was conducted for 120 min at 60 °C and pH 6.5 using Alcalase at an enzyme-substrate ratio of 1:200 w/w. The chemical refining of crude oil consisted of degumming, neutralization, washing, drying, bleaching, and deodorization; deodorization was conducted at different temperatures and processing times. Although chemical refining was successful, temperature and chemical reagents favored the removal of polyunsaturated fatty acids (PUFA) from the oil. Aroma attributes of fishy odor, frying odor, and rancid odor predominantly contributed to the sensory evaluation of the product. Deodorization conditions of 160 °C for 1h and 200 °C for 1h were recommended for the tuna by-product oil, which is rich in PUFA. Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. Optimization of enzymatic saccharification of Chaetomorpha linum biomass for the production of macroalgae-based third generation bioethanol

    Directory of Open Access Journals (Sweden)

    Ahmed Slaheddine Masmoudi

    2016-08-01

    Full Text Available To evaluate the efficacy of marine macro-algae Chaetomorpha linum as a potential biofuel resource, the effects of the enzymatic treatment conditions on sugar yield were evaluated using a three factor three level Box-Behnken design. The hydrothermally pretreated C. linum biomass was treated with Aspergillus niger cellulase at various liquid to solid ratios (50–100 mL/g, enzyme concentrations (10–60 U/g and incubations times (4–44 h. Data obtained from the response surface methodology were subjected to the analysis of variance and analyzed using a second order polynomial equation. The fitted model was found to be robust and was used to optimize the sugar yield (% during enzymatic hydrolysis. The optimum saccharification conditions were: L/S ratio 100 mL/g; enzyme concentration 52 U/g; and time 44 h. Their application led to a maximum sugar yield of 30.2 g/100g dry matter. Saccharomyces cerevisiae fermentation of the algal hydrolysate provided 8.6 g ethanol/100g dry matter. These results showed a promising future of applying C. linum biomass as potential feedstock for third generation bioethanol production.

  1. Improvements on enzymatic hydrolysis of human hair for illicit drug determination by gas chromatography/mass spectrometry.

    Science.gov (United States)

    Míguez-Framil, Martha; Moreda-Piñeiro, Antonio; Bermejo-Barrera, Pilar; López, Patricia; Tabernero, María Jesús; Bermejo, Ana María

    2007-11-15

    The use of ultrasound energy for accelerating the pronase E enzymatic hydrolysis of human hair for extracting illicit drugs has been novelty tested. The enzymatic extracts obtained after 30 min of sonication in an ultrasonic water bath were subjected to an optimized solid-phase extraction process, which involved a solution of 2.0% (v/v) acetic acid in methanol as eluting solution and concentration by N2 stream evaporation. A gas chromatography/mass spectrometry method was used to separate and determine cocaine, benzoylecgonine, codeine, morphine, and 6-monoacethylmorphine in 20 min. Variables affecting ultrasound-assisted pronase E hydrolysis such as hydrolysis temperature, hydrolysis time, enzyme concentration, catalyzer (1,4-dithiothreitol) concentration, ionic strength, pH, and ultrasound frequency were simultaneously evaluated by a Plackett-Burman design 2(8) PBD of resolution III. The most statistically significant variables were ionic strength and pH, which means that analyte extraction is mainly attributed to pronase E activity. The optimization or evaluation of all the factors has led to an accelerated pronase E hydrolysis of human hair, which can be completed in 30 min. Results have been found to be statistically similar to those obtained with conventional pronase E hydrolysis. The accelerated method was finally applied to several human hair samples from multidrug abusers.

  2. Real-time ESI-MS of enzymatic conversion: impact of organic solvents and multiplexing.

    Science.gov (United States)

    Scheerle, Romy K; Grassmann, Johanna; Letzel, Thomas

    2012-01-01

    Different enzymatic assays were characterized systematically by real-time electrospray ionization mass spectrometry (ESI-MS) in the presence of organic solvents as well as in multiplex approaches and in a combination of both. Typically, biological enzymatic reactions are studied in aqueous solutions, since most enzymes show their full activity solely in aqueous solutions. However, in recent years, the use of organic solvents in combination with enzymatic reactions has gained increasing interest due to biotechnological advantages in chemical synthesis, development of online coupled setups screening for enzyme regulatory compounds, advantages regarding mass spectrometric detection and others. In the current study, the influence of several common organic solvents (methanol, ethanol, isopropanol, acetone, acetonitrile) on enzymatic activity (hen egg white lysozyme, chitinase, α-chymotrypsin, elastase from human neutrophils and porcine pancreas, acetylcholinesterase) was tested. Moreover, multiplexing is a promising approach enabling fast and cost-efficient screening methods, e.g. for determination of inhibitors in complex mixtures or in the field of biomedical research. Although in multiplexed setups the enzymatic activity may be affected by the presence of other substrates and/or enzymes, the expected advantages possibly will predominate. To investigate those effects, we measured multiple enzymatic assays simultaneously. For all conducted measurements, the conversion rate of the substrate(s) was calculated, which reflects the enzymatic activity. The results provide an overview about the susceptibility of the selected enzymes towards diverse factors and a reference point for many applications in analytical chemistry and biotechnology.

  3. Enzymatic saccharification of dilute acid pretreated saline crops for fermentable sugar production

    Energy Technology Data Exchange (ETDEWEB)

    Zheng, Yi; Zhang, Ruihong [Biological and Agricultural Engineering Department, University of California, Davis One Shields Avenue, Davis, CA 95616 (United States); Pan, Zhongli [Biological and Agricultural Engineering Department, University of California, Davis One Shields Avenue, Davis, CA 95616 (United States); Processed Foods Research Unit, USDA-ARS-WRRC, 800 Buchanan Street, Albany, CA 94710 (United States); Wang, Donghai [Biological and Agricultural Engineering Department, Kansas State University, Manhattan, KS 66506 (United States)

    2009-11-15

    Four saline crops [athel (Tamarix aphylla L), eucalyptus (Eucalyptus camaldulensis), Jose Tall Wheatgrass (Agropyron elongatum), and Creeping Wild Ryegrass (Leymus triticoides)] that are used in farms for salt uptake from soil and drainage irrigation water have the potential for fuel ethanol production because they don't take a large number of arable lands. Dilute sulfuric acid pretreatment and enzymatic hydrolysis were conducted to select the optimum pretreatment conditions and the best saline crop for further enzymatic hydrolysis research. The optimum dilute acid pretreatment conditions included T = 165 C, t = 8 min, and sulfuric acid concentration 1.4% (w/w). Creeping Wild Ryegrass was decided to be the best saline crop. Solid loading, cellulase and {beta}-glucosidase concentrations had significant effects on the enzymatic hydrolysis of dilute acid pretreated Creeping Wild Ryegrass. Glucose concentration increased by 36 mg/mL and enzymatic digestibility decreased by 20% when the solid loading increased from 4 to 12%. With 8% solid loading, enzymatic digestibility increased by over 30% with the increase of cellulase concentration from 5 to 15 FPU/g-cellulose. Under given cellulase concentration of 15 FPU/g-cellulose, 60% increase of enzymatic digestibility of pretreated Creeping Wild Ryegrass was obtained with the increase of {beta}-glucosidase concentration up to 15 CBU/g-cellulose. With a high solid loading of 10%, fed-batch operation generated 12% and 18% higher enzymatic digestibility and glucose concentration, respectively, than batch process. (author)

  4. Optimization of enzymatic clarification of green asparagus juice using response surface methodology.

    Science.gov (United States)

    Chen, Xuehong; Xu, Feng; Qin, Weidong; Ma, Lihua; Zheng, Yonghua

    2012-06-01

    Enzymatic clarification conditions for green asparagus juice were optimized by using response surface methodology (RSM). The asparagus juice was treated with pectinase at different temperatures (35 °C-45 °C), pH values (4.00-5.00), and enzyme concentrations (0.6-1.8 v/v%). The effects of enzymatic treatment on juice clarity and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging capacity were investigated by employing a 3-factor central composite design coupled with RSM. According to response surface analysis, the optimal enzymatic treatment condition was pectinase concentration of 1.45%, incubation temperature of 40.56 °C and pH of 4.43. The clarity, juice yield, and soluble solid contents in asparagus juice were significantly increased by enzymatic treatment at the optimal conditions. DPPH radical-scavenging capacity was maintained at the level close to that of raw asparagus juice. These results indicated that enzymatic treatment could be a useful technique for producing green asparagus juice with high clarity and high-antioxidant activity. Treatment with 1.45% pectinase at 40.56 ° C, pH 4.43, significantly increased the clarity and yield of asparagus juice. In addition, enzymatic treatment maintained antioxidant activity. Thus, enzymatic treatment has the potential for industrial asparagus juice clarification. © 2012 Institute of Food Technologists®

  5. Characterization of Volatile Flavor Compounds in Chinese Rice Wine Fermented from Enzymatic Extruded Rice.

    Science.gov (United States)

    Xu, Enbo; Long, Jie; Wu, Zhengzong; Li, Hongyan; Wang, Fang; Xu, Xueming; Jin, Zhengyu; Jiao, Aiquan

    2015-07-01

    Enzymatic extrusion, instead of traditional steam cooking, to treat rice is an efficient and alternative pretreatment for Chinese rice wine fermentation. In order to determine the formation of volatiles in enzymatic extrusion-processed rice wine (EE), and to confirm its characteristic flavor compounds, headspace solid-phase micro-extraction followed by GC-MS was used. A total of 66 volatile compounds were identified in EE. During fermentation, most volatiles generated from enzymatic extruded rice had the similar trends with those from steam-cooked rice, but the differences in the concentration of volatiles indicated a changed balance of flavors release caused by enzymatic extrusion. Besides, the concentrations and sorts of volatiles in EEs fermented from different rice particle sizes, were not dramatically different. By principal component analysis, EE could be distinctly separated from other traditional Chinese rice wines according to its characteristic volatiles, namely, 2-heptanol, 1-octen-3-ol, ethyl 4-hydroxybenzoate, methylpentyl 2-propenoate, γ-hexalactone, and 4-vinylguaiacol. Enzymatic extrusion liquefaction has been a popular thermal treatment for cereals, and gradually being applied in fermentation and liquor-making industry all over the world. The characterization of volatile flavor compounds in Chinese rice wine processed by enzymatic extrusion liquefaction pretreatment, might be made use not only for a better understanding of this new-type rice wine, but for the further utilization of enzymatic extrusion in other wine or alcohol production as well. © 2015 Institute of Food Technologists®

  6. Inhibition of Lipid Peroxidation by Enzymatic Hydrolysates from Wheat Bran

    Directory of Open Access Journals (Sweden)

    Yanping Cao

    2011-01-01

    Full Text Available Wheat bran, an important by-product of the cereal industry, is rich in potentially health-promoting phenolic compounds. The phenolics are mainly esterified to the cell wall polysaccharides. In our previous paper, wheat bran was destarched and deproteinated by α-amylase, protease and amyloglucosidase successively and further hydrolyzed using Bacillus subtilis xylanases, and the enzymatic hydrolysates from wheat bran (EHWB showed good scavenging activity in vitro. The aim of this study is to further characterize the antioxidant potential of EHWB against various systems, both ex vivo and in vivo, namely, rat liver microsomal lipid peroxidation systems induced by Fe2+/H2O2 and Fe3+-adenosine diphosphate (ADP/dihydronicotinamide adenine dinucleotide phosphate (NADPH, copper- and 2,2’-azo-bis(2-amidinopropane dihydrochloride (AAPH-induced human low-density lipoprotein (LDL oxidation systems, and alloxan-induced in vivo lipid peroxidation in mice. EHWB inhibited lipid peroxidation in rat liver microsomes induced by Fe2+/H2O2 and Fe3+-ADP/NADPH in a concentration-dependent manner with 90.3 and 87 % inhibition of lipid peroxidation at 50 mg/L, respectively, which were similar to that of butylated hydroxytoluene (BHT at 20 mg/L. The antioxidant potential of EHWB at a concentration ranging from 10 to 20 mg/L in the nonenzymatic system was more effective than in the enzymatic system. EHWB strongly inhibited in vitro copper- and AAPH-mediated oxidation of LDL in a concentration- and time-dependent manner with 52.41 and 63.03 % inhibition at 20 mg/L, respectively, which were similar to that of ascorbate at 10 mg/L. EHWB significantly decreased the level of thiobarbituric acid reactive substances (TBARS and increased the activities of glutathione peroxidase (GSH-Px, catalase (CAT and superoxide dismutase (SOD in serum and liver of alloxan-treated mice compared with the control. These results demonstrated that EHWB might be efficient in the protection of

  7. Optimizing conditions for enzymatic extraction of sunflower oil

    Directory of Open Access Journals (Sweden)

    Badr, F. H.

    1992-10-01

    Full Text Available Sunflower seed oil was extracted with an enzymatic processes using different hydrolytic enzymes: cellulase, hemicellulase, animal proteinase, acid proteinase, pectinase and pectinex, as compared to enzyme - free aqueous extraction. All the hydrolytic enzymes enhanced oil extraction from sunflower seeds. The most optimal conditions for oil extraction from sunflower seeds were: 2% enzyme concentration, 30% substrate concentration and 3 hrs period. Using Boganov and Buchkov equation showed that time must be prolonged to get higher yields. The maximum yield during 3 hrs extraction with enzymatic process ranged between 44,5%-57,1% of the soxhlet extractable oil. The potency of the investigated enzymes in extracting oil was in the following order: acid proteinase > cellulase > hemicellulase > animal proteinase > pectinex > pectinase when compared at the previous optimal conditions.

    Aceite de semilla de girasol fue extraído mediante un proceso enzimático usando diferentes enzimas hidrolíticos: celulasa, hemicelulasa, proteinasa animal, proteinasa acida, pectinasa y pectinex, comparando con la extracción acuosa libre de enzima. Todos los enzimas hidrolíticos incrementan la extracción de aceites de semilla de girasol. Las condiciones óptimas para la extracción de aceite a partir de semillas de girasol fueron: 2% de concentración de enzima, 30% de concentración de sustrato y un período de 3 horas. La ecuación de Boganov y Buchkov mostró que el tiempo debe ser prolongado para alcanzar altos rendimientos. El máximo rendimiento durante tres horas de extracción con proceso enzimático osciló entre el 44,5%-57,1% del aceite extraído con soxhlet. La potencia de los enzimas investigados en la extracción de aceite siguió el orden: proteinasa acida > celulasa > hemicelulasa > proteinasa animal > pectinex > pectinasa cuando fue previamente comparado con las condiciones óptimas.

  8. Enzymatic Phorbol Esters Degradation using the Germinated Jatropha Curcas Seed Lipase as Biocatalyst: Optimization Process Conditions by Response Surface Methodology

    Directory of Open Access Journals (Sweden)

    Avita Kusuma Wardhani

    2016-10-01

    Full Text Available Utilization of Jatropha curcas seed cake is limited by the presence of phorbol esters (PE, which are the main toxic compound and heat stable. The objective of this research was to optimize the reaction conditions of the enzymatic PE degradation of the defatted Jatropha curcas seed cake (DJSC using the acetone-dried lipase from the germinated Jatropha curcas seeds as a biocatalyst. Response Surface Methodology (RSM using three-factors-three-levels Box-Behnken design was used to evaluate the effects of the reaction time, the ratio of buffer volume to DJSC, and the ratio of enzyme to DJSC on PE degradation. The results showed that the optimum conditions of PE degradation were 29.33 h, 51.11 : 6 (mL/g, and 30.10 : 5 (U/g cake for the reaction time, the ratio of buffer volume to DJSC, and the ratio of enzyme to DJSC, respectively. The predicted degradation of PE was 98.96% and not significantly different with the validated data of PE degradation. PE content was 0.035 mg/g, in which it was lower than PE in non-toxic Jatropha seeds. The results indicated that enzymatic degradation of PE might be a promising method for degradation of PE.  Copyright © 2016 BCREC GROUP. All rights reserved Received: 22nd December 2015; Revised: 1st April 2016; Accepted: 14th April 2016 How to Cite: Wardhani, A.K., Hidayat, C., Hastuti, P. (2016. Enzymatic Phorbol Esters Degradation using the Germinated Jatropha Curcas Seed Lipase as Biocatalyst: Optimization Process Conditions by Response Surface Methodology. Bulletin of Chemical Reaction Engineering & Catalysis, 11 (3: 346-353 (doi:10.9767/bcrec.11.3.574.346-353 Permalink/DOI: http://doi.org/10.9767/bcrec.11.3.574.346-353

  9. Haematology, genotoxicity, enzymatic activity and histopathology as biomarkers of metal pollution in the shrew Crocidura russula

    International Nuclear Information System (INIS)

    Sanchez-Chardi, A.; Marques, C.C.; Gabriel, S.I.; Capela-Silva, F.; Cabrita, A.S.; Lopez-Fuster, M.J.; Nadal, J.; Mathias, M.L.

    2008-01-01

    Haematological (WBC, RBC, Hgb and Hct) and genotoxicity (MNT) parameters, hepatic enzymatic activities (GST, GPx and GR), and a histopathological evaluation of liver, kidneys and gonads were assessed as general biomarkers of metal pollution in the shrew Crocidura russula inhabiting a pyrite mining area. Specimens exposed to metals presented a few significant alterations when compared with reference animals: GST activity decreased; micronuclei increased; and evident liver alterations related to metal exposure were observed. On the basis of all the parameters studied, age was an important factor that partly explained the observed variation, whereas sex was the least important factor. Significant correlations were also found between heavy metal concentrations and biomarkers evaluated, demonstrating the great influence of these metals in the metabolic alterations. To the best of our knowledge, these data constitute the first measurements of a battery of biomarkers in shrews from a mine site and are among the few available for insectivorous mammals. - Metals from an abandoned pyrite mine produce alterations in haematological parameters, GST, MNT, and histopathology in shrews

  10. Photosynthetic and enzymatic metabolism of Schinus terebinthifolius Raddi seedlings under water deficit

    Directory of Open Access Journals (Sweden)

    Danieli Pieretti Nunes

    Full Text Available ABSTRACT Schinus terebinthifolius Raddi is a tree species that can be used in the recovery of degraded areas, as it exhibits rapid growth and has a very expansive root system, facilitating water uptake from the deeper layers of the soil. The objective of this study was to evaluate photosynthesis and enzymatic activity in S. terebinthifolius seedlings under conditions of water deficit and their potential to recover following re-irrigation. The experiment was conducted in a greenhouse under a plastic covering where plants were distributed into two groups: Group 1 - control plants, where irrigation was maintained at 70% of the water retention capacity, and Group 2 - stressed plants, where irrigation was suspended until the photosynthetic rate neared zero, followed by rehydration for 12 days, then a further suspension of irrigation. At the beginning of the experiment and during the suspension of irrigation and rehydration, plants were evaluated for gas and antioxidant enzyme exchanges. Hydric stress significantly reduced photosynthesis, stomatal transpiration conductance, carboxylation efficiency of Rubisco, and the chlorophyll content of the S. terebinthifolius plants. Following rehydration, plants recovered the carboxylation efficiency of Rubisco, but not the photosynthetic rate. Antioxidant enzyme activity increased in both the aerial part and the root in response to water deficit.

  11. Production of xylooligosaccharide from wheat bran by microwave assisted enzymatic hydrolysis.

    Science.gov (United States)

    Wang, Tseng-Hsing; Lu, Shin

    2013-06-01

    The effective production of xylooligosaccharides (XOS) from wheat bran was investigated. Wheat bran contains rich hemicellulose which can be hydrolyzed by enzyme; the XOS were obtained by microwave assisted enzymatic hydrolysis. To improve the productivity of XOS, repeated microwave assisted enzymatic hydrolysis and activated carbon adsorption method was chosen to eliminate macromolecules in the XOS. On the basis of experimental data, an industrial XOS production process consisting of pretreatment, repeated microwave assisted enzymatic treatment and purification was designed. Using the designed process, 3.2g dry of purified XOS was produced from 50 g dry wheat bran powder. Copyright © 2012 Elsevier Ltd. All rights reserved.

  12. Pretreatment by radiation and acids of chaff and its effect on enzymatic hydrolysis of cellulose

    International Nuclear Information System (INIS)

    Kumakura, M.; Kaetsu, I.

    1984-01-01

    The effect of pretreatment by radiation and acids—sulfuric, hydrochloric and acetic—on the enzymatic hydrolysis of chaff was studied. The combination of radiation and acids accelerates subsequent crushing and enzymatic hydrolysis. The percentage of fine powder below 115 mesh, after the crushing and the glucose yield on subsequent enzymatic hydrolysis, increased with increasing acid concentration, treatment time and irradiation dose. Radiation and hydrochloric acid pretreatment was the most effective in giving a high glucose conversion yield (about 90%). Irradiation dose, acid concentration, treatment temperature and treatment time were 20 Mrad, 0·5%, 70°C, and 5 h, respectively

  13. Ship-borne measurements of microbial enzymatic activity: A rapid biochemical indicator for microbial water quality monitoring

    Science.gov (United States)

    Stadler, Philipp; Loken, Luke; Crawford, John; Schramm, Paul; Sorsa, Kirsti; Kuhn, Catherine; Savio, Domenico; Striegl, Rob; Butman, David; Stanley, Emily; Farnleitner, Andreas H.; Zessner, Matthias

    2017-04-01

    for ship-borne measurements of physico-chemical parameters, such as the FLAMe (Crawford et al., 2015), paves new ground for data interpretation and process understanding. Cabral, J.P.S., 2010. Water Microbiology. Bacterial Pathogens and Water. Int. J. Environ. Res. Public. Health 7, 3657-3703. doi:10.3390/ijerph7103657 Crawford, J.T., Loken, L.C., Casson, N.J., Smith, C., Stone, A.G., Winslow, L.A., 2015. High-speed limnology: using advanced sensors to investigate spatial variability in biogeochemistry and hydrology. Environ. Sci. Technol. 49, 442-450. doi:10.1021/es504773x Farnleitner, A. h., Hocke, L., Beiwl, C., Kavka, G. c., Zechmeister, T., Kirschner, A. k. t., Mach, R. l., 2001. Rapid enzymatic detection of Escherichia coli contamination in polluted river water. Lett. Appl. Microbiol. 33, 246-250. doi:10.1046/j.1472-765x.2001.00990.x Farnleitner, A.H., Hocke, L., Beiwl, C., Kavka, G.G., Mach, R.L., 2002. Hydrolysis of 4-methylumbelliferyl-β-d-glucuronide in differing sample fractions of river waters and its implication for the detection of fecal pollution. Water Res. 36, 975-981. doi:10.1016/S0043-1354(01)00288-3 Ryzinska-Paier, G., Lendenfeld, T., Correa, K., Stadler, P., Blaschke, A.P., Mach, R.L., Stadler, H., Kirschner, A.K.T., Farnleitner, A.H., 2014. A sensitive and robust method for automated on-line monitoring of enzymatic activities in water and water resources. Water Sci. Technol. J. Int. Assoc. Water Pollut. Res. 69, 1349-1358. doi:10.2166/wst.2014.032 Stadler, P., Blöschl, G., Vogl, W., Koschelnik, J., Epp, M., Lackner, M., Oismüller, M., Kumpan, M., Nemeth, L., Strauss, P., Sommer, R., Ryzinska-Paier, G., Farnleitner, A.H., Zessner, M., 2016. Real-time monitoring of beta-d-glucuronidase activity in sediment laden streams: A comparison of prototypes. Water Res. 101, 252-261. doi:10.1016/j.watres.2016.05.072 Stadler, P., Farnleitner, A.H., Zessner, M., 2017. Development and evaluation of a self-cleaning custom-built auto sampler controlled by a low

  14. OPTIMIZATION OF CHEMICAL AND ENZYMATIC DEINKING OF PHOTOCOPIER WASTE PAPER

    Directory of Open Access Journals (Sweden)

    Puneet Pathak

    2011-02-01

    Full Text Available The utilization of post-consumer papers in the production of new paper products is increasing all over the world in recent years. Recycling of photocopier paper is a major problem due to difficulty in removal of non-impact ink. Enzymes offer potential advantages in ecofriendly deinking of recovered paper. In this study the deinking of photocopier paper was examined using chemicals and a commercial cellulase enzyme. Parameters of deinking experiments were optimized for hydrapulping. The ink was removed by flotation and washing processes. Then these parameters were compared in terms of ink removal ability of the process, as well as optical and strength properties of the deinked paper. The application of enzymatic deinking improved ink removal efficiency by 24.6% and freeness by 21.6% with a reduction in drainage time of 11.5% in comparison to those obtained with chemical deinking. The physical properties, namely burst index and tensile index, were observed to improve by 15.3% and 2.7%, respectively and brightness and tear index decreased by 2.1% and 21.9%, respectively. Results of deinking efficiency of photocopier paper showed that the enzyme used in the present work performed better than the conventional chemicals used for deinking.

  15. ENZYMATIC BIODIESEL SYNTHESIS FROM ACID OIL USING A LIPASE MIXTURE

    Directory of Open Access Journals (Sweden)

    Kelly C. N. R. Pedro

    Full Text Available The conventional biodiesel production process has some disadvantages. It is necessary to use refined vegetable oils with low free fatty acids (FFAs content. An alternative route is to use low-cost acid oils in an enzymatic process. The use of lipases allows simultaneous esterification of FFAs and transesterification of triglycerides present in raw material forming alkyl esters. The aim of this work was to study the production of biodiesel using soybean oils with different acid contents (Acid Value of 8.5, 50, 90 and ethanol catalyzed by commercial immobilized lipases (Novozym 435, Lipozyme RM IM and Lipozyme TL IM. A significant decrease of acid value was observed mainly with Novozym 435 and Lipozyme RM IM. The use of a mixture of two immobilized lipases was also investigated to decrease catalyst cost and increase the amount of ester produced. The three commercial immobilized lipases were mixed in a dual system and tested for biodiesel synthesis from acid oil (AV of 8.5, 50 and 90. A positive synergistic effect occurred mainly for Lipozyme TL IM (1,3-specific lipase and Novozym 435 (non-specific lipase blend. The ester content doubled when this lipase mixture was used in ethanolysis of acid oil with AV of 90.

  16. A novel chemo-enzymatic synthesis of hydrophilic phytosterol derivatives.

    Science.gov (United States)

    He, Wen-Sen; Hu, Di; Wang, Yu; Chen, Xue-Yan; Jia, Cheng-Sheng; Ma, Hai-Le; Feng, Biao

    2016-02-01

    In this study, a novel method was developed for chemo-enzymatic synthesis of hydrophilic phytosterol derivatives, phytosteryl polyethylene glycol succinate (PPGS), through an intermediate phytosteryl hemisuccinate (PSHS), which was first chemically prepared and subsequently coupled with polyethylene glycol (PEG) through lipase-catalyzed esterification. The chemical structure of intermediate and goal product were finally confirmed to be PSHS and PPGS by FT-IR, MS and NMR, suggesting that hydrophilic phytosterol derivatives were successfully synthesized. The effects of various parameters on the conversion of PSHS to PPGS were investigated and the highest conversion (>78%) was obtained under the selected conditions: 75 mmol/L PSHS, 1:2M ratio of PSHS to PEG, 50 g/L Novozym 435, 120 g/L 3 Å molecular sieves in tert-butanol, 55 °C, 96 h and 200 rpm. The solubility of phytosterols in water was significantly improved by coupling with PEG, facilitating the incorporation into a variety of foods containing water. Copyright © 2015 Elsevier Ltd. All rights reserved.

  17. Industrial enzymatic production of cephalosporin-based beta-lactams.

    Science.gov (United States)

    Barber, Michael S; Giesecke, Ulrich; Reichert, Arno; Minas, Wolfgang

    2004-01-01

    Cephalosporins are chemically closely related to penicillins both work by inhibiting the cell wall synthesis of bacteria. The first generation cephalosporins entered the market in 1964. Second and third generation cephalosporins were subsequently developed that were more powerful than the original products. Fourth generation cephalosporins are now reaching the market. Each newer generation of cephalosporins has greater Gram-negative antimicrobial properties than the preceding generation. Conversely, the 'older' generations of cephalosporins have greater Gram-positive (Staphylococcus and Streptococcus) coverage than the 'newer' generations. Frequency of dosing decreases and palatability generally improve with increasing generations. The advent of fourth generation cephalosporins with the launch of cefepime extended the spectrum against Gram-positive organisms without a significant loss of activity towards Gram-negative bacteria. Its greater stability to beta-lactamases increases its efficacy against drug-resistant bacteria. In this review we present the current situation of this mature market. In addition, we present the current state of the technologies employed for the production of cephalosporins, focusing on the new and environmentally safer 'green' routes to the products. Starting with the fermentation and purification of CPC, enzymatic conversion in conjunction with aqueous chemistry will lead to some key intermediates such as 7-ACA, TDA and TTA, which then can be converted into the active pharmaceutical ingredient (API), again applying biocatalytic technologies and aqueous chemistry. Examples for the costing of selected products are provided as well.

  18. Enzymatic Mercury Detoxification: The Regulatory Protein MerR

    CERN Multimedia

    Ctortecka, B; Walsh, C T; Comess, K M

    2002-01-01

    Mercury ions and organomercurial reagents are extremely toxic due to their affinity for thiol groups. Many bacteria contain an elaborate detoxification system for a metabolic conversion of toxic Hg$^{2+}$ or organomercurials to less toxic elemental Hg$^0$. The main components of the enzymatic mercury detoxification (see Fig. 1) are the regulatory protein MerR (mercury responsive genetic switch), the organomercurial lyase MerB (cleavage of carbon mercury bonds), and the mercuric ion reductase MerA (reduction of mercuric ions). In these proteins Hg$^{2+}$ is usually coordinated by the thiol groups of cysteines. We utilize the nuclear quadrupole interaction (NQI) of ${\\rm^{199m}}$Hg detected by time differential perturbed angular correlation (TDPAC) to identify the Hg metal site geometries in these proteins in order to elucidate the molecular origin of the ultrasensitivity, selectivity and reaction mechanism of this detoxification system. The short lived TDPAC probe ${\\rm^{199m}}$Hg ($\\tau_{1/2} =$ 43 min) is su...

  19. Enzymatic desulfation of the red seaweeds agar by Marinomonas arylsulfatase.

    Science.gov (United States)

    Wang, Xueyan; Duan, Delin; Fu, Xiaoting

    2016-12-01

    Agar and sulfated galactans were isolated from the red seaweeds Gracilariopsis lemaneiformis and Gelidium amansii. A previously purified arylsulfatase from Marinomonas sp. FW-1 was used to remove sulfate groups in agar and sulfated galactans. After enzymatic desulfation, the sulfate content decreased to about 0.16% and gel strength increased about two folds. Moreover, there was no difference between the DNA electrophoresis spectrum on the gel of the arylsulfatase-treated agar and that of the commercial agarose. In order to reveal the desulfation ratio and site, chemical and structural identification of sulfated galactan were carried out. G. amansii sulfated galactan with 7.4% sulfated content was composed of galactose and 3,6-anhydro-l-galactose. Meanwhile, G. lemaneiformis sulfated galactan with 8.5% sulfated content was composed of galactose, 3,6-anhydro-l-galactose, 2-O-methyl-3,6-anhydro-l-galactose and xylose. Data from 13 C NMR, FT-IR, GC-MS provided evidence of sulfate groups at C-4 and C-6 of d-galactose and C-6 of l-galactose both in GRAP and GEAP. Data from GC-MS revealed that desulfation was carried out by the arylsulfatase at the sulfate bonds at C-4 and C-6 of d-galactose and C-6 of l-galactose, with a desulfation ratio of 83.4% and 86.0% against GEAP and GRAP, respectively. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. Modeling the minimum enzymatic requirements for optimal cellulose conversion

    International Nuclear Information System (INIS)

    Den Haan, R; Van Zyl, W H; Van Zyl, J M; Harms, T M

    2013-01-01

    Hydrolysis of cellulose is achieved by the synergistic action of endoglucanases, exoglucanases and β-glucosidases. Most cellulolytic microorganisms produce a varied array of these enzymes and the relative roles of the components are not easily defined or quantified. In this study we have used partially purified cellulases produced heterologously in the yeast Saccharomyces cerevisiae to increase our understanding of the roles of some of these components. CBH1 (Cel7), CBH2 (Cel6) and EG2 (Cel5) were separately produced in recombinant yeast strains, allowing their isolation free of any contaminating cellulolytic activity. Binary and ternary mixtures of the enzymes at loadings ranging between 3 and 100 mg g −1 Avicel allowed us to illustrate the relative roles of the enzymes and their levels of synergy. A mathematical model was created to simulate the interactions of these enzymes on crystalline cellulose, under both isolated and synergistic conditions. Laboratory results from the various mixtures at a range of loadings of recombinant enzymes allowed refinement of the mathematical model. The model can further be used to predict the optimal synergistic mixes of the enzymes. This information can subsequently be applied to help to determine the minimum protein requirement for complete hydrolysis of cellulose. Such knowledge will be greatly informative for the design of better enzymatic cocktails or processing organisms for the conversion of cellulosic biomass to commodity products. (letter)

  1. Enzymatic Halogenation and Dehalogenation Reactions: Pervasive and Mechanistically Diverse.

    Science.gov (United States)

    Agarwal, Vinayak; Miles, Zachary D; Winter, Jaclyn M; Eustáquio, Alessandra S; El Gamal, Abrahim A; Moore, Bradley S

    2017-04-26

    Naturally produced halogenated compounds are ubiquitous across all domains of life where they perform a multitude of biological functions and adopt a diversity of chemical structures. Accordingly, a diverse collection of enzyme catalysts to install and remove halogens from organic scaffolds has evolved in nature. Accounting for the different chemical properties of the four halogen atoms (fluorine, chlorine, bromine, and iodine) and the diversity and chemical reactivity of their organic substrates, enzymes performing biosynthetic and degradative halogenation chemistry utilize numerous mechanistic strategies involving oxidation, reduction, and substitution. Biosynthetic halogenation reactions range from simple aromatic substitutions to stereoselective C-H functionalizations on remote carbon centers and can initiate the formation of simple to complex ring structures. Dehalogenating enzymes, on the other hand, are best known for removing halogen atoms from man-made organohalogens, yet also function naturally, albeit rarely, in metabolic pathways. This review details the scope and mechanism of nature's halogenation and dehalogenation enzymatic strategies, highlights gaps in our understanding, and posits where new advances in the field might arise in the near future.

  2. Enzymatic synthesis of 13N-β-nicotinamide adenine dinucleotide

    International Nuclear Information System (INIS)

    Lambrecht, R.H.D.; Slegers, G.; Claeys, A.; Vandecasteele, C.

    1985-01-01

    Nitrogen-13-labelled β-nicotinamide adenine dinucleotide ( 13 N-NAD) is an interesting new compound for positron emission tomography. A semi-automatic production method is developed that yields a solution of 13 N-NAD of radiopharmaceutical quality, suitable for human intravenous administration. The 13 N-NAD is prepared enzymatically in one step from cyclotron-produced 13 NH 3 and nicotinic acid adenine dinucleotide (deamido-NAD). The enzyme NAD synthetase (E.C. 6.3.1.5), catalysing this reaction, is extracted and purified from Escherichia coli. The purified enzyme is immobilized by glutaraldehyde coupling to γ-aminopropylsilane-coated porous glass beads. The enzyme-loaded glass beads are packed in a column. The kinetic properties of the column are optimized. For synthetizing 13 N-NAD, the mixture of co-factors and substrates, containing 13 NH 3 , is pumped over the enzyme column. The unreacted 13 NH 3 is separated from 13 N-NAD by on-line passage over a cation exchanger. After passing over a millipore filter, a sterile solution of radiochemically pure 13 N-NAD is obtained, containing 70 mCi in 10 mL. The total synthesis time is 10 minutes. The specific activity is about 120 mCi/μmol at EOB. Quality control includes sterility and pyrogen tests, HPLC and HPTLC analysis. (author)

  3. Enzymatically and chemically oxidized lignin nanoparticles for biomaterial applications.

    Science.gov (United States)

    Mattinen, Maija-Liisa; Valle-Delgado, Juan José; Leskinen, Timo; Anttila, Tuomas; Riviere, Guillaume; Sipponen, Mika; Paananen, Arja; Lintinen, Kalle; Kostiainen, Mauri; Österberg, Monika

    2018-04-01

    Cross-linked and decolorized lignin nanoparticles (LNPs) were prepared enzymatically and chemically from softwood Kraft lignin. Colloidal lignin particles (CLPs, ca. 200 nm) in a non-malodorous aqueous dispersion could be dried and redispersed in tetrahydrofuran (THF) or in water retaining their stability i.e. spherical shape and size. Two fungal laccases, Trametes hirsuta (ThL) and Melanocarpus albomyces (MaL) were used in the cross-linking reactions. Reactivity of ThL and MaL on Lignoboost™ lignin and LNPs was confirmed by high performance size exclusion chromatography (HPSEC) and oxygen consumption measurements with simultaneous detection of red-brown color due to the formation of quinones. Zeta potential measurements verified oxidation of LNPs via formation of surface-oriented carboxylic acid groups. Dynamic light scattering (DLS) revealed minor changes in the particle size distributions of LNPs after laccase catalyzed radicalization, indicating preferably covalent intraparticular cross-linking over polymerization. Changes in the surface morphology of laccase treated LNPs were imaged by atomic force (AFM) and transmission emission (TEM) microscopy. Furthermore, decolorization of LNPs without degradation was obtained using ultrasonication with H 2 O 2 in alkaline reaction conditions. The research results have high impact for the utilization of Kraft lignin as nanosized colloidal particles in advanced bionanomaterial applications in medicine, foods and cosmetics including different sectors from chemical industry. Copyright © 2018 The Author(s). Published by Elsevier Inc. All rights reserved.

  4. Enzymatic synthesis of rubber polymer in Hevea brasiliensis

    Energy Technology Data Exchange (ETDEWEB)

    Tang, F.; Hu, S.; Benedict, C.R. (Texas A and M Univ., College Station (United States))

    1991-05-01

    Light and Dennis purified serum soluble rubber transferase from Hevea latex to homogeneity. Prenyl transferase co-purified with rubber transferase. In the absence of washed rubber particles (WRP) the prenyl transferase catalyzed the formation of trans FPP from DMAPP and IPP. In the presence of WRP the transferase catalyzed cis additions of IPP to pre-existing rubber chains. Control mixtures of WRP, Mg{sup 2+} and FPP were not included to test for the contributions of the bound rubber transferase on WRP to the incorporation of IPP into polyisoprene. Bound rubber transferase catalyzes the repetitive addition of IPP to allylic-PP starter molecules to form polyisoprene. The order of utilization of allylic-PP starters was GGPP > FPP > GPP > DMAPP. The authors have shown that the polyisoprene enzymatically synthesized on WRP is a bimodal polymer consisting of different mol wt rubber chains similar to the polymeric characteristics of natural rubber. The bound rubber transferase was solubilized with Chaps and purified on DEAE-cellulose. The polymerization reaction catalyzed by the purified preparation showed a 98% requirement for pre-existing rubber chains. Results suggest that the prenyl transferase from Hevea serum may be part of the polymer starter system furnishing allylic-PP for the bound rubber transferase.

  5. Multiphoton manipulations of enzymatic photoactivity in aspartate aminotransferase.

    Science.gov (United States)

    Hill, Melissa P; Freer, Lucy H; Vang, Mai C; Carroll, Elizabeth C; Larsen, Delmar S

    2011-04-21

    The aspartate aminotransferase (AAT) enzyme utilizes the chromophoric pyridoxal 5'-phosphate (PLP) cofactor to facilitate the transamination of amino acids. Recently, we demonstrated that, upon exposure to blue light, PLP forms a reactive triplet state that rapidly (in microseconds) generates the high-energy quinonoid intermediate when bound to PLP-dependent enzymes [J. Am. Chem. Soc.2010, 132 (47), 16953-16961]. This increases the net catalytic activity (k(cat)) of AAT, since formation of the quinonoid is partially rate limiting via the thermally activated enzymatic pathway. The magnitude of observed photoenhancement initially scales linearly with pump fluence; however when a critical threshold is exceeded, the photoactivity saturates and is even suppressed at greater excitation fluences. The photodynamic mechanisms associated with this suppression behavior are characterized with the use of ultrafast multipulse pump-dump-probe and pump-repump-probe transient absorption techniques in combination with complementary two-color, steady-state excitation assays. Via multistate kinetic modeling of the transient ultrafast data and the steady-state assay data, the nonmonotonic incident power dependence of the photoactivty in AAT is decomposed into contributions from high-intensity dumping of the excited singlet state and repumping of the excited triplet state with induces the repopulation of the ground state via rapid intersystem crossing in the higher-lying triplet electronic manifold.

  6. Highly Branched Bio-Based Unsaturated Polyesters by Enzymatic Polymerization

    Directory of Open Access Journals (Sweden)

    Hiep Dinh Nguyen

    2016-10-01

    Full Text Available A one-pot, enzyme-catalyzed bulk polymerization method for direct production of highly branched polyesters has been developed as an alternative to currently used industrial procedures. Bio-based feed components in the form of glycerol, pentaerythritol, azelaic acid, and tall oil fatty acid (TOFA were polymerized using an immobilized Candida antarctica lipase B (CALB and the potential for an enzymatic synthesis of alkyds was investigated. The developed method enables the use of both glycerol and also pentaerythritol (for the first time as the alcohol source and was found to be very robust. This allows simple variations in the molar mass and structure of the polyester without premature gelation, thus enabling easy tailoring of the branched polyester structure. The postpolymerization crosslinking of the polyesters illustrates their potential as binders in alkyds. The formed films had good UV stability, very high water contact angles of up to 141° and a glass transition temperature that could be controlled through the feed composition.

  7. Enzymatic carotenoid cleavage in star fruit (Averrhoa carambola).

    Science.gov (United States)

    Fleischmann, Peter; Watanabe, Naoharu; Winterhalter, Peter

    2003-05-01

    This paper presents the first description of an enzyme fraction exhibiting carotenoid cleavage activity isolated from fruit skin of Averrhoa carambola. Partial purification of the enzyme could be achieved by acetone precipitation, ultrafiltration (300 kDa, 50 kDa), isoelectric focusing (pH 3-10) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (7.5%). In this way, an enzymatically active protein fraction was obtained, consisting of four proteins in the molecular weight range of between 12 and 90 kDa. Using beta-carotene as substrate, the enzyme activity was detected spectrophotometrically at 505 nm. The main reaction product, detected by GC analysis, was beta-ionone. This proves that the isolated enzymes are closely related to aroma metabolism and release of star fruit. The time constant of the reaction was 16.6 min, the Michaelis Constant K(m)=3.6 micromol 1(-1) and the maximum velocity V(max)=10.5 x 10(-3) micromol l(-1) s(-1) mg((Protein))(-1). The optimum temperature was 45 degrees C.

  8. One-step process of hydrothermal and alkaline treatment of wheat straw for improving the enzymatic saccharification.

    Science.gov (United States)

    Sun, Shaolong; Zhang, Lidan; Liu, Fang; Fan, Xiaolin; Sun, Run-Cang

    2018-01-01

    To increase the production of bioethanol, a two-step process based on hydrothermal and dilute alkaline treatment was applied to reduce the natural resistance of biomass. However, the process required a large amount of water and a long operation time due to the solid/liquid separation before the alkaline treatment, which led to decrease the pure economic profit for production of bioethanol. Therefore, four one-step processes based on order of hydrothermal and alkaline treatment have been developed to enhance concentration of glucose of wheat straw by enzymatic saccharification. The aim of the present study was to systematically evaluated effect for different one-step processes by analyzing the physicochemical properties (composition, structural change, crystallinity, surface morphology, and BET surface area) and enzymatic saccharification of the treated substrates. In this study, hemicelluloses and lignins were removed from wheat straw and the morphologic structures were destroyed to various extents during the four one-step processes, which were favorable for cellulase absorption on cellulose. A positive correlation was also observed between the crystallinity and enzymatic saccharification rate of the substrate under the conditions given. The surface area of the substrate was positively related to the concentration of glucose in this study. As compared to the control (3.0 g/L) and treated substrates (11.2-14.6 g/L) obtained by the other three one-step processes, the substrate treated by one-step process based on successively hydrothermal and alkaline treatment had a maximum glucose concentration of 18.6 g/L, which was due to the high cellulose concentration and surface area for the substrate, accompanying with removal of large amounts of lignins and hemicelluloses. The present study demonstrated that the order of hydrothermal and alkaline treatment had significant effects on the physicochemical properties and enzymatic saccharification of wheat straw. The one

  9. Recyclability of PET/WPI/PE Multilayer Films by Removal of Whey Protein Isolate-Based Coatings with Enzymatic Detergents

    Directory of Open Access Journals (Sweden)

    Patrizia Cinelli

    2016-06-01

    yield, stress and elongation at break, evaluated by tensile testing on films before and after cleaning, were are not significantly affected by washing with enzymatic detergents.

  10. Recyclability of PET/WPI/PE Multilayer Films by Removal of Whey Protein Isolate-Based Coatings with Enzymatic Detergents

    Science.gov (United States)

    Cinelli, Patrizia; Schmid, Markus; Bugnicourt, Elodie; Coltelli, Maria Beatrice; Lazzeri, Andrea

    2016-01-01

    elongation at break, evaluated by tensile testing on films before and after cleaning, were are not significantly affected by washing with enzymatic detergents. PMID:28773592

  11. Metabolite profiling of enzymatically hydrolyzed and fermented forms of Opuntia ficus-indica and their effect on UVB-induced skin photoaging.

    Science.gov (United States)

    Cho, Dong-Woon; Kim, Dae-Eung; Lee, Dae-Hee; Jung, Kyung-Hoon; Hurh, Byung-Serk; Kwon, Oh Wook; Kim, Sun Yeou

    2014-01-01

    Fermentation of natural products is emerging as an important processing method and is attracting a lot of attention because it may have the advantage of having a new biological function. In this study, fruits of Opuntia ficus-indica were enzymatically hydrolyzed and then fermented with two species of yeast. We identified novel prominent markers in enzymatically hydrolyzed O. ficus-indica (EO) and fermented O. ficus-indica (FO) samples by using an ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry. We also evaluated the effect of EO and FO on photoaging of skin cells exposed to ultraviolet radiation. We identified the major fermented metabolite in the FO as ferulic acid. Our in vitro study indicated that FO significantly enhanced the concentration of pro-collagen type 1 than the EO, by increasing the TGF-β1 production.

  12. Physico-chemical, spectroscopical and thermal characterization of bio diesel obtained by enzymatic route as a tool to select the most efficient immobilized lipase

    International Nuclear Information System (INIS)

    Silva, G.A.M.; Ros, P.C.M. da; Souza, L.T.A.; Costa, A.P.O.; Castro, H.F. de

    2012-01-01

    Two microbial lipases from Burkholderia cepacia and Pseudomonas fluorescens were evaluated as catalysts for the enzymatic transesterification of beef tallow with ethanol and the most efficient lipase source was selected by taking into account the properties of the product to be used as fuel. Both lipases were immobilized on an epoxy silica-polyvinyl alcohol composite by covalent immobilization and used to perform the reactions under the following operational conditions: beef tallow-to-ethanol molar ratio of 1:9, 45 degree C and 400 units of enzymatic activity per gram of fat. Products, characterized using Fourier Transform Infrared spectroscopy (FTIR), viscosimetry, thermogravimetry and 1 H NMR spectroscopy, suggested that the bio diesel sample obtained in the reaction catalyzed by Burkholderia cepacia lipase has the best set of properties for fuel usage. (author)

  13. Physico-chemical, spectroscopical and thermal characterization of bio diesel obtained by enzymatic route as a tool to select the most efficient immobilized lipase

    Energy Technology Data Exchange (ETDEWEB)

    Silva, G.A.M.; Ros, P.C.M. da; Souza, L.T.A.; Costa, A.P.O.; Castro, H.F. de, E-mail: heizir@dequi.eel.usp.br [Engineering School of Lorena. University of Sao Paulo (EEL/USP), Lorena, SP (Brazil)

    2012-01-15

    Two microbial lipases from Burkholderia cepacia and Pseudomonas fluorescens were evaluated as catalysts for the enzymatic transesterification of beef tallow with ethanol and the most efficient lipase source was selected by taking into account the properties of the product to be used as fuel. Both lipases were immobilized on an epoxy silica-polyvinyl alcohol composite by covalent immobilization and used to perform the reactions under the following operational conditions: beef tallow-to-ethanol molar ratio of 1:9, 45 degree C and 400 units of enzymatic activity per gram of fat. Products, characterized using Fourier Transform Infrared spectroscopy (FTIR), viscosimetry, thermogravimetry and {sup 1}H NMR spectroscopy, suggested that the bio diesel sample obtained in the reaction catalyzed by Burkholderia cepacia lipase has the best set of properties for fuel usage. (author)

  14. Influence of enzymatic hydrolysis on the allergenic reactivity of processed cashew and pistachio.

    Science.gov (United States)

    Cuadrado, Carmen; Cheng, Hsiaopo; Sanchiz, Africa; Ballesteros, Isabel; Easson, Michael; Grimm, Casey C; Dieguez, M Carmen; Linacero, Rosario; Burbano, Carmen; Maleki, Soheila J

    2018-02-15

    Cashew and pistachio allergies are considered a serious health problem. Previous studies have shown that thermal processing, pressurization and enzymatic hydrolysis may reduce the allergenic properties of food by changing the protein structure. This study assesses the allergenic properties of cashew and pistachio after thermal treatment (boiling and autoclaving), with or without pressure (autoclaving), and multiple enzymatic treatments under sonication, by SDS-PAGE, western blot and ELISA, with serum IgE of allergic individuals, and mass spectroscopy. Autoclaving and enzymatic hydrolysis under sonication separately induced a measurable reduction in the IgE binding properties of pastes made from treated cashew and pistachio nuts. These treatments were more effective with pistachio allergens. However, heat combined with enzymatic digestion was necessary to markedly lower IgE binding to cashew allergens. The findings identify highly effective simultaneous processing conditions to reduce or even abolish the allergenic potency of cashew and pistachio. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. Numerical prediction of kinetic model for enzymatic hydrolysis of cellulose using DAE-QMOM approach

    Science.gov (United States)

    Jamil, N. M.; Wang, Q.

    2016-06-01

    Bioethanol production from lignocellulosic biomass consists of three fundamental processes; pre-treatment, enzymatic hydrolysis, and fermentation. In enzymatic hydrolysis phase, the enzymes break the cellulose chains into sugar in the form of cellobiose or glucose. A currently proposed kinetic model for enzymatic hydrolysis of cellulose that uses population balance equation (PBE) mechanism was studied. The complexity of the model due to integrodifferential equations makes it difficult to find the analytical solution. Therefore, we solved the full model of PBE numerically by using DAE-QMOM approach. The computation was carried out using MATLAB software. The numerical results were compared to the asymptotic solution developed in the author's previous paper and the results of Griggs et al. Besides confirming the findings were consistent with those references, some significant characteristics were also captured. The PBE model for enzymatic hydrolysis process can be solved using DAE-QMOM method. Also, an improved understanding of the physical insights of the model was achieved.

  16. Switchgrass storage effects on the recovery of carbohydrates after liquid hot water pretreatment and enzymatic hydrolysis

    Directory of Open Access Journals (Sweden)

    Danielle Julie Carrier

    2016-08-01

    Full Text Available Perennial grasses that would be used for bioenergy and bioproducts production will need to be stored for various periods of time to ensure a continual feedstock supply to a bioprocessing facility. The effects of storage practices on grass composition and the response of grasses to subsequent bioprocesses such as pretreatment and enzymatic hydrolysis needs to be understood to develop the most efficient storage protocols. This study examined the effect of outdoor storage of round switchgrass bales on composition before and after liquid hot water pretreatment (LHW and enzymatic hydrolysis. This study also examined the effect of washing LHW pretreated biomass prior to enzymatic hydrolysis. It was determined that switchgrass composition after baling was stable. As expected, glucan and lignin contents increased after LHW due to decreases in xylan and galactan. Washing biomass prior to enzymatic hydrolysis reduced saccharification, especially in samples from the interior of the bale, by at least 5%.

  17. Rapid near infrared spectroscopy for prediction of enzymatic hydrolysis of corn bran after various pretreatments

    DEFF Research Database (Denmark)

    Baum, Andreas; Wittrup Agger, Jane; Meyer, Anne S.

    2012-01-01

    Efficient generation of a fermentable hydrolysate is a primary requirement in the utilization of fibrous plant biomass as feedstocks in bioethanol processes. The first biomass conversion step usually involves a hydrothermal pretreatment before enzymatic hydrolysis. The purpose of the pretreatment...... step is to increase the responsivity of the substrate to enzymatic attack and the type of pretreatment affects the enzymatic conversion efficiency. Destarched corn bran is a fibrous, heteroxylan-rich side-stream from the starch industry which may be used as a feedstock for bioethanol production...... release of different levels of arabinose, xylose and glucose from all the differently pretreated destarched corn bran samples. The present study also demonstrates a generic, non-destructive solution to determine the enzymatic monosaccharide release from polymers in biomass side-streams, thereby...

  18. Enzymatic browning control in cut apples (Red delicious through a system of active packaging

    Directory of Open Access Journals (Sweden)

    Felipe Jadán Piedra

    2017-03-01

    Full Text Available Enzymatic browning is one of the most relevant mechanisms of deterioration that take place in fresh-cut fruit and vegetables, as a consequence of the activity of the polyphenol oxidase enzyme on the phenolic compounds release after cellular lysis . This work is focused on the reduction of these enzymatic activity by an active packaging technology, which make use of a material that incorporates antioxidant active agents. Thus, films of ethylene-vynil alcohol copolymer (EVOH containing a typical food antioxidant, such as ascorbic acid and a polyphenol oxidase-inhibiting agent, the 4-hexylresorcinol have been developed and used to wrap apple slices. The evolution of color, the enzymatic activity and the kinetic of agents release to food simulants were monitored. The results showed an improvement of apple slice color stability and a reduction of the enzymatic activity. The film with 10 % of agents in 3/1 ratio (4-hexylresorcinol/ascorbic acid provided the best results.

  19. Enzymatic vegetable organic extracts as soil biochemical biostimulants and atrazine extenders.

    Science.gov (United States)

    García-Martínez, Ana María; Tejada, Manuel; Díaz, Ana Isabel; Rodríguez-Morgado, Bruno; Bautista, Juan; Parrado, Juan

    2010-09-08

    The purpose of this study was to gather information on the potential effects of organic biostimulants on soil activity and atrazine biodegradation. Carob germ enzymatic extract (CGEE) and wheat condensed distiller solubles enzymatic extract (WCDS-EE) have been obtained using an enzymatic process; their main organic components are soluble carbohydrates and proteins in the form of peptides and free amino acids. Their application to soil results in high biostimulation, rapidly increased dehydrogenase, phosphatase and glucosidase activities, and an observed atrazine extender capacity due to inhibition of its mineralization. The extender capacity of both extracts is proportional to the protein/carbohydrate ratio content. As a result, these enzymatic extracts are highly microbially available, leading to two independent phenomena, fertility and an atrazine persistence that is linked to increased soil activity.

  20. Method of reduction of nitroaromatics by enzymatic reaction with redox enzymes

    Science.gov (United States)

    Shah, Manish M.

    2000-01-01

    A method for the controlled reduction of nitroaromatic compounds such as nitrobenzene and 2,4,6-trinitrotoluene by enzymatic reaction with redox enzymes, such as Oxyrase (Trademark of Oxyrase, Inc., Mansfield, Ohio).