Search results for: ligninolytic enzyme

Authors: Debora B. Moretti, Wiolene M. Nordi, Thaline Maira P. Cruz, José Eurico P. Cyrino, Raul Machado-Neto

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Enzyme activity was evaluated in the intestine of juvenile dourado (Salminus brasiliensis) fed with diets containing 0, 10 or 20% of lyophilized bovine colostrum (LBC) inclusion for either 30 or 60 days. The intestinal enzymes acid and alkaline phosphatase (ACP and ALP, respectively), non-specific esterase (NSE), lipase (LIP), dipeptidyl aminopeptidase IV (DAP IV) and leucine aminopeptidase (LAP) were studied using histochemistry in four intestinal segments (S1, S2, S3 and posterior intestine). Weak proteolitic activity was observed in all intestinal segments for DAP IV and LAP. The activity of NSE and LIP was also weak in all intestines, except for the moderate activity of NSE in the S2 of 20% LBC group after 30 days and in the S1 of 0% LBC group after 60 days. The ACP was detected only in the S2 and S3 of the 10% LBC group after 30 days. Moderate and strong staining was observed in the first three intestinal segments for ALP and weak activity in the posterior intestine. The activity of DAP IV, LAP and ALP were also present in the cytoplasm of the enterocytes. In the present results, bovine colostrum feeding did not cause alterations in activity of intestinal enzymes.

Keywords: Carnivorous fish, enterocyte, intestinal epithelium, teleost.

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Authors: Ahmad M. Khalil, Ahmad M. Alshamali, Marwan H. Gagaa

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We investigated oxidative DNA damage caused by radio frequency radiation using 8-oxo-7, 8-dihydro-2'- deoxyguanosine (8-oxodG) generated in mice tissues after exposure to 900 MHz mobile phone radio frequency in three independent experiments. The RF was generated by a Global System for Mobile Communication (GSM) signal generator. The radio frequency field was adjusted to 25 V/m. The whole body specific absorption rate (SAR) was 1.0 W/kg. Animals were exposed to this field for 30 min daily for 30 days. 24 h post-exposure, blood serum, brain and spleen were removed and DNA was isolated. Enzyme-linked immunosorbent assay (ELISA) was used to measure 8-oxodG concentration. All animals survived the whole experimental period. The body weight of animals did not change significantly at the end of the experiment. No statistically significant differences observed in the levels of oxidative stress. Our results are not in favor of the hypothesis that 900 MHz RF induces oxidative damage.

Keywords: Mice, Mobile phone radiation, oxidative stress, 8-oxo-7, 8-dihydro-2'-deoxyguanosine

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Authors: Ivan Živanović, Žužana Vaštag, Senka Popović, Ljiljana Popović, Draginja Peričin

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The present work represents an investigation of the hydrolysis of hull-less pumpkin (Cucurbita Pepo L.) oil cake protein isolate (PuOC PI) by pepsin. To examine the effectiveness and suitability of pepsin towards PuOC PI the kinetic parameters for pepsin on PuOC PI were determined and then, the hydrolysis process was studied using Response Surface Methodology (RSM). The hydrolysis was carried out at temperature of 30°C and pH 3.00. Time and initial enzyme/substrate ratio (E/S) at three levels were selected as the independent parameters. The degree of hydrolysis, DH, was mesuared after 20, 30 and 40 minutes, at initial E/S of 0.7, 1 and 1.3 mA/mg proteins. Since the proposed second-order polynomial model showed good fit with the experimental data (R2 = 0.9822), the obtained mathematical model could be used for monitoring the hydrolysis of PuOC PI by pepsin, under studied experimental conditions, varying the time and initial E/S. To achieve the highest value of DH (39.13 %), the obtained optimum conditions for time and initial E/S were 30 min and 1.024 mA/mg proteins.

Keywords: Enzymatic hydrolysis, Pepsin, Pumpkin (CucurbitaPepo L.) oil cake protein isolate, Response surface methodology.

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Authors: Noraziah A. Y., Mashitah M. D., Subhash Bhatia

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Fructooligosaccharides derived from microbial enzyme especially from fungal sources has been received particular attention due to its beneficial effects as prebiotics and mass production. However, fungal fermentation is always cumbersome due to its broth rheology problem that will eventually affect the production of FOS. This study investigated the efficiency of immobilized cell system using rotating fibrous bed bioreactor (RFBB) in producing fructooligosaccharides (FOS). A comparative picture with respect to conventional stirred tank bioreactor (CSTB) and RFBB has been presented. To demonstrate the effect of agitation intensity and aeration rate, a laboratory-scale bioreactor 2.5 L was operated in three phases (high, medium, low) for 48 hours. Agitation speed has a great influence on P. simplicissimum fermentation for FOS production, where the volumetric FOS productivity using RFBB is increased with almost 4 fold compared to the FOS productivity in CSTB that only 0.319 g/L/h. Rate of FOS production increased up to 1.2 fold when immobilized cells system was employed at aeration rate similar to the freely suspended cells at 2.0 vvm.

Keywords: Fructooligosaccharides, immobilized, productivity, prebiotics.

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Authors: Hala O. El-Mesallamy, Salwa M. Suwailem, Mae M. Seleem

Abstract:

Visfatin and apelin are two new adipokines that recently gained a special interest in diabetes research. This study was conducted to study the interplay between these two adipokines and their correlation with other inflammatory and biochemical parameters in type 2 diabetic (T2D) postmenopausal women with CAD. Visfatin and apelin were measured by enzyme-linked immunoassay (ELISA). Visfatin was found to be significantly higher in the following groups: T2D patients without CAD, non-obese and obese T2D patients with CAD when compared to control group. Apelin was found to be significantly lower in non-obese and obese T2D patients with CAD when compared to control group. Visfatin and apelin were found to be significantly associated with each other and with other biochemical parameters. The current study provides evidence for the interplay between visfatin and apelin through the inflammatory milieu characteristic of T2D and their possible role in the pathogenesis of CAD complication of T2D. 

Keywords: Apelin, Coronary artery disease, Inflammation, Type 2 diabetes, Visfatin.

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Authors: Sang-Ho Baik

Abstract:

D-erythro-cyclohexylserine (D chiral unnatural β-hydroxy amino acid expected for the synthesis of drug for AIDS treatment. To develop a continuous bioconversion system with whole cell biocatalyst of D-threonine aldolase (D genes for the D-erythro-CHS production, D-threonine aldolase gene was amplified from Ensifer arboris 100383 by direct PCR amplication using two degenerated oligonucleotide primers designed based on genomic sequence of Shinorhizobium meliloti Sequence analysis of the cloned DNA fragment revealed one open-reading frame of 1059 bp and 386 amino acids. This putative D-TA gene was cloned into NdeI and EcoRI (pEnsi His-tag sequence or BamHI (pEnsi-DTA[2]) sequence of the pET21(a) vector. The expression level of the cloned gene was extremely overexpressed by E. coli BL21(DE3) transformed with pEnsi-DTA[1] compared to E. coli BL21(DE3) transformed with pEnsi-DTA[2]. When the cells expressing the wild used for D-TA enzyme activity, 12 mM glycine was successfully detected in HPLC analysis. Moreover, the whole cells harbouring the recombinant D-TA was able to synthesize D-erythro of 0.6 mg/ml in a batch reaction.

Keywords: About four key words or phrases in alphabetical order, separated by commas.

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Authors: R. Haddad, K. Morris, V. Buchanan-Wollaston

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Biochemical and molecular analysis of some antioxidant enzyme genes revealed different level of gene expression on oilseed (Brassica napus). For molecular and biochemical analysis, leaf tissues were harvested from plants at eight different developmental stages, from young to senescence. The levels of total protein and chlorophyll were increased during maturity stages of plant, while these were decreased during the last stages of plant growth. Structural analysis (nucleotide and deduced amino acid sequence, and phylogenic tree) of a complementary DNA revealed a high level of similarity for a family of Catalase genes. The expression of the gene encoded by different Catalase isoforms was assessed during different plant growth phase. No significant difference between samples was observed, when Catalase activity was statistically analyzed at different developmental stages. EST analysis exhibited different transcripts levels for a number of other relevant antioxidant genes (different isoforms of SOD and glutathione). The high level of transcription of these genes at senescence stages was indicated that these genes are senescenceinduced genes.

Keywords: Biochemical analysis, Oilseed, Expression pattern, Growth phases

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Authors: S. K. Mohan, T. Viruthagiri, C. Arunkumar

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Tannase (tannin acyl hydrolase, E.C.3.1.1.20) is an important hydrolysable enzyme with innumerable applications and industrial potential. In the present study, a kinetic model has been developed for the batch fermentation used for the production of tannase by A.flavus MTCC 3783. Maximum tannase activity of 143.30 U/ml was obtained at 96 hours under optimum operating conditions at 35oC, an initial pH of 5.5 and with an inducer tannic acid concentration of 3% (w/v) for a fermentation period of 120 hours. The biomass concentration reaches a maximum of 6.62 g/l at 96 hours and further there was no increase in biomass concentration till the end of the fermentation. Various unstructured kinetic models were analyzed to simulate the experimental values of microbial growth, tannase activity and substrate concentration. The Logistic model for microbial growth , Luedeking - Piret model for production of tannase and Substrate utilization kinetic model for utilization of substrate were capable of predicting the fermentation profile with high coefficient of determination (R2) values of 0.980, 0.942 and 0.983 respectively. The results indicated that the unstructured models were able to describe the fermentation kinetics more effectively.

Keywords: Aspergillus flavus, Batch fermentation, Kinetic model, Tannase, Unstructured models.

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Authors: Nahid A. Ali, Salma H. Ahmed, ElShazali A. Mohamed, Isam A. Mohamed Ahmed, Elfadil E.Babiker

Abstract:

The effect of cross linking of the protein isolates of three legumes with the microbial enzyme transglutaminase (EC 2.3.2.13) on the functional properties at different NaCl concentration was studied. The reduction in the total free amino groups (OD340) of the polymerized protein showed that TGase treatment cross-linking the protein subunit of each legume. The solubility of the protein polymer of each legume was greatly improved at high concentration of NaCl. At 1.2 M NaCl the solubility of the native legumes protein was significantly decreased but after polymerization slightly improved. Cross linked proteins were less turbid on heating to higher temperature as compared to native proteins and the temperature at which the protein turns turbid also increased in the polymerized proteins. The emulsifying and foaming properties of the protein polymer were greatly improved at all concentrations of NaCl for all legumes.

Keywords: Functional properties, Legumes, Protein isolate, NaCl, Transglutaminase.

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Authors: Chanin Park, Sundarapandian Thangapandian, Yuno Lee, Minky Son, Shalini John, Young-sik Sohn, Keun Woo Lee

Abstract:

Hypertension is characterized with stress on the heart and blood vessels thus increasing the risk of heart attack and renal diseases. The Renin angiotensin system (RAS) plays a major role in blood pressure control. Renin is the enzyme that controls the RAS at the rate-limiting step. Our aim is to develop new drug-like leads which can inhibit renin and thereby emerge as therapeutics for hypertension. To achieve this, molecular dynamics (MD) simulation and receptor-based pharmacophore modeling were implemented, and three rennin-inhibitor complex structures were selected based on IC50 value and scaffolds of inhibitors. Three pharmacophore models were generated considering conformations induced by inhibitor. The compounds mapped to these models were selected and subjected to drug-like screening. The identified hits were docked into the active site of renin. Finally, hit1 satisfying the binding mode and interaction energy was selected as possible lead candidate to be used in novel renin inhibitors.

Keywords: Renin inhibitor, Molecular dynamics simulation, Structure-based pharmacophore modeling

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Authors: S.H. Mirdamadian, S.M. Khayam-Nekoui, H. Ghanavati

Abstract:

Composting is the process in which municipal solid waste (MSW) and other organic waste materials such as biosolids and manures are decomposed through the action of bacteria and other microorganisms into a stable granular material which, applied to land, as soil conditioner. Microorganisms, especially those that are able to degrade polymeric organic material have a key role in speed up this process. The aim of this study has been established to isolation of microorganisms with high ability to production extracellular enzymes for degradation of natural polymers that are exists in MSW for decreasing time of degradation phase. Our experimental study for isolation designed in two phases: in first phase we isolated degrading microorganism with selected media that consist a special natural polymer such as cellulose, starch, lipids and etc as sole source of carbon. In second phase we selected microorganism that had high degrading enzyme production with enzymatic assay for seed production. However, our findings in pilot scale have indicated that usage of this microbial consortium had high efficiency for decreasing degradation phase.

Keywords: Biodegradation, Compost, Municipal Solid Waste, Waste Management.

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Authors: J. Carlos Kuri, Varun Vij, Raymond J. Turner, Orly Yadid-Pecht

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Celiac disease (CD) is an immune system disorder that is related to eating gluten. As gluten-free (GF) diet has become a concern of many people for health reasons, a gold standard had to be nominated. Enzyme-linked immunosorbent assay (ELISA) has taken the seat of this role. However, multiple limitations were discovered, and with that, the desire for an alternative method now exists. Nucleic acid based aptamers have become of great interest due to their selectivity, specificity, simplicity, and rapid-testing advantages. However, fluorescence-based aptasensors have been tagged as unstable, but lifespan details are rarely stated. In this work, the lifespan stability of a fluorescence-based aptasensor is shown over a 8-week long study displaying the accuracy of the sensor and false negatives. This study follows 22 different samples, including GF and gluten-rich (GR) and soy sauce products, off-the-shelf products, and reference material from laboratories; giving a total of 836 tests. The analysis shows an accuracy of correctly classifying GF and GR products of 96.30% and 100%, respectively, when the protocol is augmented with molecular sieves. The overall accuracy remains around 94% within the first 4 weeks and then decays to 63%.

Keywords: Aptasensor, PEG, rGO, FAM, RM, ELISA.

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Authors: Feng Chia Hsieh, Sheng Kuo Hsieh, Tzyy Rong Jinn

Abstract:

Characterization and evaluation of the activity of Vespa basalis DPP-IV, which expressed in Spodoptera frugiperda 21 cells. The expression of rDPP-IV was confirmed by SDS–PAGE, Western blot analyses, LC-MS/MS and measurement of its peptidase specificity. One-step purification by Ni-NTA affinity chromatography and the total amount of rDPP-IV recovered was approximately 6.4mg per liter from infected culture medium; an equivalent amount would be produced by 1x109 infected Sf21 insect cells. Through the affinity purification led to highly stable rDPP-IV enzyme was recovered and with significant peptidase activity. The rDPP-IV exhibited classical Michaelis–Menten kinetics, with kcat/Km in the range of 10-500 mM-1×S-1 for the five synthetic substrates and optimum substrate is Ala-Pro-pNA. As expected in inhibition assay, the enzymatic activity of rDPP-IV was significantly reduced by 80 or 60% in the presence of sitagliptin (a DPP-IV inhibitor) or PMSF (a serine protease inhibitor), but was not apparently affected by iodoacetamide (a cysteine protease inhibitor).

Keywords: Dipeptidyl-Peptidase IV, Phenylmethylsulfonyl fluoride; Serine protease, Sitagliptin, Vespa basalis

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Authors: M. Maleki, M. Latifi, M. Amani-Tehran

Abstract:

Nano fibers produced by electrospinning are of industrial and scientific attention due to their special characteristics such as long length, small diameter and high surface area. Applications of electrospun structures in nanotechnology are included tissue scaffolds, fibers for drug delivery, composite reinforcement, chemical sensing, enzyme immobilization, membrane-based filtration, protective clothing, catalysis, solar cells, electronic devices and others. Many polymer and ceramic precursor nano fibers have been successfully electrospun with diameters in the range from 1 nm to several microns. The process is complex so that fiber diameter is influenced by various material, design and operating parameters. The objective of this work is to apply genetic algorithm on the parameters of electrospinning which have the most significant effect on the nano fiber diameter to determine the optimum parameter values before doing experimental set up. Effective factors including initial polymer concentration, initial jet radius, electrical potential, relaxation time, initial elongation, viscosity and distance between nozzle and collector are considered to determine finest diameter which is selected by user.

Keywords: Electrospinning, genetic algorithm, nano fiber diameter, optimization.

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Authors: Ishtiaq Ahmed, Muhammad Anjum Zia, Hafiz Muhammad Nasir Iqbal

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The purpose of the present work was to study the production and process parameters optimization for the synthesis of cellulase from Trichoderma viride in solid state fermentation (SSF) using an agricultural wheat straw as substrates; as fungal conversion of lignocellulosic biomass for cellulase production is one among the major increasing demand for various biotechnological applications. An optimization of process parameters is a necessary step to get higher yield of product. Several kinetic parameters like pretreatment, extraction solvent, substrate concentration, initial moisture content, pH, incubation temperature and inoculum size were optimized for enhanced production of third most demanded industrially important cellulase. The maximum cellulase enzyme activity 398.10±2.43 μM/mL/min was achieved when proximally analyzed lignocellulosic substrate wheat straw inocubated at 2% HCl as pretreatment tool along with distilled water as extraction solvent, 3% substrate concentration 40% moisture content with optimum pH 5.5 at 45°C incubation temperature and 10% inoculum size.

Keywords: Cellulase, Lignocellulosic residue, Processoptimization, Proximal analysis, SSF, Trichoderma viride.

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Authors: S. Manzoor, A. Nadeem, M. Javed, ME. Babar

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A major goal in animal genetics is to understand the role of common genetic variants in diseases susceptibility and production traits. Sahiwal cattle can be considered as a global animal genetic resource due to its relatively high milk producing ability, resistance against tropical diseases and heat tolerant. CYP11B1 gene provides instructions for making a mitochondrial enzyme called steroid 11-beta-hydroxylase. It catalyzes the 11deoxy-cortisol to cortisol and 11deoxycorticosterone to corticosterone in cattle. The bovine CYP11B1 gene is positioned on BTA14q12 comprises of eight introns and nine exons and protein is associated with mitochondrial epithelium. The present study was aimed to identify the single-nucleotide polymorphisms in CYP11B1 gene in Sahiwal cattle breed of Pakistan. Four polymorphic sites were identified in exon one of CYP11B1 gene through sequencing approach. Significant finding was the incidence of the C→T polymorphism in 5'-UTR, causing amino acid substitution from alanine to valine (A30V) in Sahiwal cattle breed. That Ala/Val polymorphism may serve as a powerful genetic tool for the development of DNA markers that can be used for the particular traits for different local cattle breeds.

Keywords: CYP11B1, single nucleotide polymorphism, sahiwal cattle, Pakistan.

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Authors: Natasha Hussain, Maleeha Aslam, Robina Farooq

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Hepatitis B and hepatitis C are among the most significant hepatic infections all around the world that may lead to hepatocellular carcinoma. This study is first time performed at the blood transfussion centre of Omar hospital, Lahore. It aims to determine the sero-prevalence of these diseases by screening the apparently healthy blood donors who might be the carriers of HBV or HCV and pose a high risk in the transmission. It also aims the comparison between the sensitivity of two diagnostic tests; chromatographic immunoassay – one step test device and Enzyme Linked Immuno Sorbant Assay (ELISA). Blood serum of 855 apparently healthy blood donors was screened for Hepatitis B surface antigen (HBsAg) and for anti HCV antibodies. SPSS version 12.0 and X2 (Chi-square) test were used for statistical analysis. The seroprevalence of HCV was 8.07% by the device method and by ELISA 9.12% and that of HBV was 5.6% by the device and 6.43% by ELISA. The unavailability of vaccination against HCV makes it more prevalent. Comparing the two diagnostic methods, ELISA proved to be more sensitive.

Keywords: ELISA, Sensitivity comparison of diagnostic tests, seroprevalence of Hepatitis B and C

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Authors: K. Obikeze, P. Mugabo, I. Green, D. Dietrich, A. Burger

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Leonotisleonurus a shrub indigenous to Southern Africa is widely used in traditional medicine to treat a variety of conditions ranging from skin diseases and cough to epileptic fits and ‘heart problems’. Studies on the aqueous extract of the leaves have indicated cycloxegenase enzyme inhibitory activity and an antihypertensive effect. Five methanol leaf extract fractions (MLEa - MLEe) of L. leonurus were tested on anaesthetized normotensive male Wistar rats (AWR) and isolated perfused working rat hearts (IWH). Fraction MLEc (0.01mg/kg – 0.05mg/kg) induced significant increases in BP and HR in AWR and positive chronotropic and inotropic effects in IWH (1.0mg/ml – 5.0mg/ml). Pre-administration of atenolol (2.0mg/kg) and prazosin (60μg/kg) significantly inhibited MLEc effect on HR and MAP respectively in vivo, while atenolol (7.0mg/ml) pre-perfusion significantly inhibited MLEc effect in vitro. The hypertensive effect of MLEc is probably via β1agonism. Results also indicate the presence of multiple cardioactive compounds in L. leonurus.

Keywords: Cardiovascular effect, in vitro, in vivo, isolated perfused working heart, Leonotis leonurus, rat.

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Authors: Minky Son, Chanin Park, Ayoung Baek, Shalini John, Keun Woo Lee

Abstract:

3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) catalyzes the conversion of HMG-CoA to mevalonate using NADPH and the enzyme is involved in rate-controlling step of mevalonate. Inhibition of HMGR is considered as effective way to lower cholesterol levels so it is drug target to treat hypercholesterolemia, major risk factor of cardiovascular disease. To discover novel HMGR inhibitor, we performed structure-based pharmacophore modeling combined with molecular dynamics (MD) simulation. Four HMGR inhibitors were used for MD simulation and representative structure of each simulation were selected by clustering analysis. Four structure-based pharmacophore models were generated using the representative structure. The generated models were validated used in virtual screening to find novel scaffolds for inhibiting HMGR. The screened compounds were filtered by applying drug-like properties and used in molecular docking. Finally, four hit compounds were obtained and these complexes were refined using energy minimization. These compounds might be potential leads to design novel HMGR inhibitor.

Keywords: Anti-hypercholesterolemia drug, HMGR inhibitor, Molecular dynamics simulation, Structure-based pharmacophore modeling.

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Authors: A. Doosti, P. Ghasemi Dehkordi, M. Zamani, S. Taheri, M. Banitalebi, M. Mahmoudzadeh

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The p53 tumor suppressor gene plays two important roles in genomic stability: blocking cell proliferation after DNA damage until it has been repaired, and starting apoptosis if the damage is too critical. Codon 72 exon4 polymorphism (Arg72Pro) of the P53 gene has been implicated in cancer risk. Various studies have been done to investigate the status of p53 at codon 72 for arginine (Arg) and proline (Pro) alleles in different populations and also the association of this codon 72 polymorphism with various tumors. Our objective was to investigate the possible association between P53 Arg72Pro polymorphism and susceptibility to colorectal cancer among Isfahan and Chaharmahal Va Bakhtiari (a part of south west of Iran) population. We investigated the status of p53 at codon 72 for Arg/Arg, Arg/Pro and Pro/Pro allele polymorphisms in blood samples from 145 colorectal cancer patients and 140 controls by Nested-PCR of p53 exon 4 and digestion with BstUI restriction enzyme and the DNA fragments were then resolved by electrophoresis in 2% agarose gel. The Pro allele was 279 bp, while the Arg allele was restricted into two fragments of 160 and 119 bp. Among the 145 colorectal cancer cases 49 cases (33.79%) were homozygous for the Arg72 allele (Arg/Arg), 18 cases (12.41%) were homozygous for the Pro72 allele (Pro/Pro) and 78 cases (53.8%) found in heterozygous (Arg/Pro). In conclusion, it can be said that p53Arg/Arg genotype may be correlated with possible increased risk of this kind of cancers in south west of Iran.

Keywords: TP53, Polymorphism, Colorectal Cancer, Iran

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Authors: Jaya Mahar Maligan, Teti Estiasih, Joni Kusnadi

Abstract:

Structured phospholipids from commercial soybean lecithin with oil enriched omega-3 fatty acid form by product of tuna canning is alternative procedure to provides the stability of omega-3 fatty acid structure and increase these bioactive function in metabolism. Best treatment condition was obtain in 18 hours acidolysis reaction with 30% enzyme concentration, which EPADHA incorporation level was 127,47 mg/g and incorporation percentage of EPA-DHA was 51,04% at phospholipids structure. This structured phospolipids could reduce atherosclerosis risk in male Sprague dawley rat. Provision of structured phospholipids has significant effect (α = 0.05) on changes in lipid profile, intima-media thickness of aorta rats (male Sprague dawley) fed atherogenic diet. Structured phospholipids intake can lower total cholesterol 78.36 mg/dL, total triglycerides 94,57 mg/dL, LDL levels 87.08 mg/dL and increased HDL level as much as 12,64 mg/dL in 10 weeks cares. Structured phospholipids intake also can prevent the thickening of the intima-media layer of the aorta.

Keywords: Structured phospholipids, commercial soybean lecithin, omega-3 fatty acid, atherosclerosis risk.

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Authors: Rafayel A. Azizyan, Aram E. Gevorgyan, Valeri B. Arakelyan, Emil S. Gevorgyan

Abstract:

Currently, mathematical and computer modeling are widely used in different biological studies to predict or assess behavior of such a complex systems as a biological are. This study deals with mathematical and computer modeling of bi-substrate enzymatic reactions, which play an important role in different biochemical pathways. The main objective of this study is to represent the results from in silico investigation of bi-substrate enzymatic reactions in the presence of uncompetitive inhibitors, as well as to describe in details the inhibition effects. Four models of uncompetitive inhibition were designed using different software packages. Particularly, uncompetitive inhibitor to the first [ES₁] and the second ([ES₁S₂]; [FS₂]) enzyme-substrate complexes have been studied. The simulation, using the same kinetic parameters for all models allowed investigating the behavior of reactions as well as determined some interesting aspects concerning influence of different cases of uncompetitive inhibition. Besides, it has been shown that uncompetitive inhibitors exhibit specific selectivity depending on mechanism of bi-substrate enzymatic reaction. 

Keywords: Mathematical modeling, bi-substrate enzymatic reactions, sequential mechanism, ping-pong mechanism, uncompetitive inhibition.

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Authors: J. Salaenoi, A. Thongpan, M. Mingmuang

Abstract:

The activities of alkaline phosphatase and Ca2+ATPase in mud crab (Scylla serrata) collected from a soft-shell crab farm in Chantaburi Province, Thailand, in several stages of molting cycle were observed. The results showed that the activity of alkaline phosphatase in gill after molting was highly significant (p<0.05) comparing to those at intermolt and premolt stages. The activity profiles of alkaline phosphatase in integument and haemolymph were similar showing a decrease from intermolt to 2- week premolt stage and increased during 2-day premolt to 6-h postmolt stage before dropping at 7-day postmolt stage, while this enzyme in the gill was quite low at intermolt and premolt stages. For Ca2+ATPase, the activity profiles in gill and integument corresponded to the molting variation, especially the activities increased during 5-7 day postmolt stage were at highly significant levels (p<0.05) comparing to those at premolt and early postmolt stages. The highest activity of Ca2+ATPase in haemolymph was found at 2-week premolt stage (p<0.05). Changes in alkaline phosphatase and Ca2+ATPase activities over the molting cycle clearly indicated their active functions on calcification.

Keywords: Scylla serrata, molting cycle, alkaline phosphatase, Ca2+ATPase

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Authors: Glauber Cruz, Patrícia A. S. Monteiro, Carlos E. M. Braz, Paulo Seleghin Jr., Igor Polikarpov, Paula M.Crnkovic

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Enzymatic hydrolysis is one of the major steps involved in the conversion from sugarcane bagasse to yield ethanol. This process offers potential for yields and selectivity higher, lower energy costs and milder operating conditions than chemical processes. However, the presence of some factors such as lignin content, crystallinity degree of the cellulose, and particle sizes, limits the digestibility of the cellulose present in the lignocellulosic biomasses. Pretreatment aims to improve the access of the enzyme to the substrate. In this study sugarcane bagasse was submitted chemical pretreatment that consisted of two consecutive steps, the first with dilute sulfuric acid (1 % (v/v) H2SO4), and the second with alkaline solutions with different concentrations of NaOH (1, 2, 3 and 4 % (w/v)). Thermal Analysis (TG/ DTG and DTA) was used to evaluate hemicellulose, cellulose and lignin contents in the samples. Scanning Electron Microscopy (SEM) was used to evaluate the morphological structures of the in natura and chemically treated samples. Results showed that pretreatments were effective in chemical degradation of lignocellulosic materials of the samples, and also was possible to observe the morphological changes occurring in the biomasses after pretreatments.

Keywords: Alkaline solutions, bioethanol production, dilute acid, enzymatic hydrolysis, lignocellulosic biomass.

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Authors: Ashish Kumar Nayak, Dhamodharan K., Ajay S. Kalamdhad

Abstract:

The present study is aimed at alteration of sewage sludge into stable compost product using vermicomposting of sewage sludge mixed with cattle manure and saw dust in five different proportions based on C/N ratios (C/N 15 (R1), 20 (R2), 25 (R3) and 30 (R4); and control (R5)) by employing an epigeic earthworm Eisenia fetida. Higher reductions in C/N ratio, CO₂ evolution and OUR were observed in R4 demonstrated the compost stability. In addition, R4 proved to be best combination for the growth of the earthworms. In order to observe the optimal degradation, kinetics for degradation of organic matter in vermicomposting were quantitatively evaluated. An approach model was developed by assuming that composting process is carried out in a homogeneous way and the kinetics for decomposition reaction is represented by a Monod-type equation. The results exhibit comparable variations in the kinetic constants K_m and K₃ under varying parameters during vermicomposting process. Results suggested that higher R² value in R4, enhanced suitability towards Lineweaver-Burke plot. R4 yields higher degradability coefficient (K) reveals that the occurrence of optimal nutrient balance, which not only enhanced the affinity of enzymes towards substrate but also improved its degradation process. Therefore, it can be proved that R4 provided to be the best feed combination for vermicomposting process as compared to other reactors.

Keywords: Vermicomposting, Eisenia fetida, Sewage sludge, C/N ratio, Stability, Enzyme kinetics concept.

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Authors: P. Thongmuang, Y. Sudjaroen

Abstract:

The methanolic extracts from seeds of tamarind (Tamarindus indica) was prepared by Soxhlet apparatus extraction and evaluated for total phenolic content by Folin-Ciocalteu method. Then, methanolic extract was screened biological activities (In vitro) for anti-melanogenic activity by tyrosinase inhibition test, antiinflammation activity by cyclooxygenase 1 (COX-1) and cyclooxygenase 2 (COX-2) inhibition test, and cytotoxic screening test with Vero cells. The results showed that total phenolic content, which contained in extract, was contained 27.72 mg of gallic acid equivalent per g of dry weight. The ability to inhibit tyrosinase enzyme, which exerted by Tamarind seed extracts (1 mg/ml) was 52.13 ± 0.42 %. The extract was not possessed inhibitory effect to COX-1 and COX-2 enzymes and cytotoxic effect to Vero cells. The finding is concludes that tested seed extract was possessed antimelanogenic activity with non-toxic effects. However, there was not exhibited anti-inflammatory activity. Further studies include the use of advance biological models to confirm this biological activity, as well as, the isolation and characterization of the purified compounds that it was contained.

Keywords: Tamarindus indica, anti-melanogenic, antiinflammatotion, cytotoxicity, seed, phenolic compounds.

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Authors: T. A. Abegunde, O. B. Oyewole, T. A. Sanni

Abstract:

A process of conversion of flour from three varieties of cassava, namely Odongbo, ofege and TMS30752 to ethanol using α-amylase locally sourced from germinated unhusked paddy rice and yeast isolated from palm wine was developed. It involves the germination of paddy rice for a period of 15days to produce α-amylase for starch hydrolysis and isolation of yeast from palm wine for fermentation. The results showed that optimum amylase yield of “ofada” rice paddy was at 6^th day germination which was 576.9ml/g. Ethanol yield for TMS30572 (440.3%) was significantly higher than “Odongbo” (160.2%) and “Ofege’’ (115.1%), Sugar conversion efficiency were 311.0%v/v, 268.2%v/v and 186.84%v/v for TMS30572, “Odongbo” and “Ofege” respectively. The ethanol boiling points were 78^oC, 76^oC and 80^oC for TMS30572, “Odongbo” and “Ofege” respectively. This study showed that cassava varieties affects quality of ethanol produced and germination of “ofada” rice for 6 days ensures optimum production of crude amylase enzyme.

Keywords: Cassava, ethanol, fermentation, hydrolysis, α-amylase.

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Authors: Payel Ghosh, Rama Chandra Pradhan, Sabyasachi Mishra

Abstract:

Jamun (Syzygium cuminii L.) is one of the important indigenous minor fruit with high medicinal value. The jamun cultivation is unorganized and there is huge loss of this fruit every year. The perishable nature of the fruit makes its postharvest management further difficult. Due to the strong cell wall structure of pectin-protein bonds and hard seeds, extraction of juice becomes difficult. Enzymatic treatment has been commercially used for improvement of juice quality with high yield. The objective of the study was to optimize the best treatment method for juice extraction. Enzymes (Pectinase and Tannase) from different stains had been used and for each enzyme, best result obtained by using response surface methodology. Optimization had been done on the basis of physicochemical property, nutritional property, sensory quality and cost estimation. According to quality aspect, cost analysis and sensory evaluation, the optimizing enzymatic treatment was obtained by Pectinase from Aspergillus aculeatus strain. The optimum condition for the treatment was 44 ^oC with 80 minute with a concentration of 0.05% (w/w). At these conditions, 75% of yield with turbidity of 32.21NTU, clarity of 74.39%T, polyphenol content of 115.31 mg GAE/g, protein content of 102.43 mg/g have been obtained with a significant difference in overall acceptability.

Keywords: Jamun, enzymatic treatment, physicochemical property, sensory analysis, optimization.

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Authors: Mardiati Zain, N. Jamarun, A. S. Tjakradidjaja

Abstract:

The effect of phosphorus supplementation of ammoniated rice straw was studied. The in vitro experiment was carried out following the first stage of Tilley and Terry method. The treatments consisting of four diets were A = 50% ammoniated rice straw + 50% concentrate (control), B = A + 0.2% Phosphor (P) supplement, C = A + 0.4% Phosphor (P) supplement, and D = A + 0.6% Phosphor (P) supplement of dry matter. Completely randomized design was used as the experimental design with differences among treatment means were examined using Duncan multiple range test. Variables measured were total bacterial and cellulolytic bacterial population, cellulolytic enzyme activity, ammonia (NH3) and volatile fatty acid (VFA) concentrations, as fermentability indicators and synthesized microbial protein, as well as degradability indicators including dry matter (DM), organic matter (OM), neutral detergent fibre (NDF), acid detergent fibre (ADF) and cellulose. The results indicated that fermentability and degradability of diets consisting ammoniated rice straw with P supplementation were significantly higher than the control diet (P< 0.05). It is concluded that P supplementation is important to improve fermentability and degradability of rations containing ammoniated RS and concentrate. In terms of the most effective level of P supplementation occurred at a supplementation rate of 0.4% of dry matter.

Keywords: Ammoniated rice straw, phosphorus, fermentability, degradability and synthesized microbial protein.

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Authors: Waranya Pongpaiboon, Warangkana Srichamnong, Supat Chaiyakul

Abstract:

Enzymatic modification of rice flour can produce highly functional derivatives use in food industries. This study aimed to evaluate the physical properties and resistant starch content of rice flour residues hydrolyzed by α-amylase. Rice flour hydrolyzed by α-amylase (60 and 300 u/g) for 1, 24 and 48 hours were investigated. Increasing enzyme concentration and hydrolysis time resulted in decreased rice flour residue’s lightness (L*) but increased redness (a*) and yellowness (b*) of rice flour residues. The resistant starch content and peak viscosity increased when hydrolysis time increased. Pasting temperature, trough viscosity, breakdown, final viscosity, setback and peak time of the hydrolyzed flours were not significantly different (p>0.05). The morphology of native flour was smooth without observable pores and polygonal with sharp angles and edges. However, after hydrolysis, granules with a slightly rough and porous surface were observed and a rough and porous surface was increased with increasing hydrolyzed time. The X-ray diffraction patterns of native flour showed A-type configuration, which hydrolyzed flour showed almost 0% crystallinity indicated that both amorphous and crystalline structures of starch were simultaneously hydrolyzed by α-amylase.

Keywords: α-Amylase, Enzymatic hydrolysis, Pasting properties, Resistant starch

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