Search results for: enzyme pre-treatment

Authors: M. Mydlárová Blaščáková, J. Poráčová, Z. Tomková, Ľ. Blaščáková, M. Nagy, M. Konečná, E. Petrejčíková, Z. Gogaľová, V. Sedlák, J. Mydlár, M. Zahatňanská, K. Hricová

Abstract:

Osteoporosis is a multifactorial disease that results in reduced quality of life, causes decreased bone strength, and changes in their microarchitecture. Mostly postmenopausal women are at risk. In our study, we measured anthropometric parameters of postmenopausal women (104 women of control group – CG and 105 women of osteoporotic group - OG) and determined TSH hormone levels and PTH as well as mineral elements - Ca, P, Mg and enzyme alkaline phosphatase. Through the correlation analysis in CG, we have found association based on age and BMI, P and Ca, as well as Mg and Ca; in OG we determined interdependence based on an association of age and BMI, age and Ca. Using the Student's t test, we found significantly important differences in biochemical parameters of Mg (p ˂ 0,001) and TSH (p ˂ 0,05) between CG and OG.

Keywords: Factors, bone mass density, Central Europe, biomarkers.

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Authors: P. Chaijak, M. Lertworapreecha, C. Sukkasem

Abstract:

A huge of dark color palm oil mill effluent (POME) cannot pass the discharge standard. It has been identified as the major contributor to the pollution load into ground water. Here, lignin-degrading yeast isolated from a termite nest was tested to treat the POME. Its lignin-degrading and decolorizing ability was determined. The result illustrated that Galactomyces sp. was successfully grown in POME. The decolorizing test demonstrated that 40% of Galactomyces sp. could reduce the color of POME (50% v/v) about 74-75% in 5 days without nutrient supplement. The result suggested that G. reessii has a potential to apply for decolorizing the dark wastewater like POME and other industrial wastewaters.

Keywords: Decolorization, palm oil mill effluent, ligninolytic enzyme, yeast, termite.

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Authors: Ž. Vaštag, Lj. Popović, S. Popović

Abstract:

After cold pressing of pumpkin oil, the defatted oil cake (PUOC) was utilised as raw material for processing of bio-functional hydrolysates. In this study, the in vitro bioactivity of an alcalase (AH) and a pepsin hydrolysate (PH) prepared from the major pumpkin 12S globulin (cucurbitin) are compared. The hydrolysates were produced at optimum reaction conditions (temperature, pH) for the enzymes, during 60min. The bioactivity testing included antioxidant and angiotensin I converting enzyme inhibitory activity assays. The hydrolysates showed high potential as natural antioxidants and possibly antihypertensive agents in functional food or nutraceuticals. Additionally, preliminary studies have shown that both hydrolysates could exhibit modest α-amylase inhibitory activity, which indicates on their hypoglycemic potential.

Keywords: Cucurbitin, alcalase, pepsin, protein hydrolysates, in vitro bioactivity.

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Authors: Anabel Fernandez, Leandro Rodriguez-Ortiz, Rosa Rodríguez

Abstract:

Due to the negative impact of fossil fuels, renewable energies are promising sources to limit global temperature rise and damage to the environment. Also, the development of technology is focused on obtaining energetic products from renewable sources. In this study, a thermodynamic model including exergy balance and a subsequent Life Cycle Assessment (LCA) were carried out for four subsystems of the integrated gasification-combustion of pinewood. Results of exergy analysis and LCA showed the process feasibility in terms of exergy efficiency and global energy efficiency of the life cycle (GEELC). Moreover, the energy return on investment (EROI) index was calculated. The global exergy efficiency resulted in 67%. For pretreatment, reaction, cleaning, and electric generation subsystems, the results were 85%, 59%, 87%, and 29%, respectively. Results of LCA indicated that the emissions from the electric generation caused the most damage to the atmosphere, water, and soil. GEELC resulted in 31.09% for the global process. This result suggested the environmental feasibility of an integrated gasification-combustion system. EROI resulted in 3.15, which determines the sustainability of the process.

Keywords: Exergy analysis, Life Cycle Assessment, LCA, renewability, sustainability.

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Authors: Reihaneh Ahmadzadeh Ghavidel, Mehdi Ghiafeh Davoodi

Abstract:

Dehydration process was carried out for tomato slices of var. Avinash after giving different pre-treatments such as calcium chloride (CaCl2), potassium metabisulphite (KMS), calcium chloride and potassium metabisulphite (CaCl2 +KMS), and sodium chloride (NaCl). Untreated samples served as control. Solar drier and continuous conveyor (tunnel) drier were used for dehydration. Quality characteristics of tomato slices viz. moisture content, sugar, titratable acidity, lycopene content, dehydration ratio, rehydration ratio and non-enzymatic browning as affected by dehydration process were studied. Storage study was also carried out for a period of six months for tomato powder packed into different types of packaging materials viz. metalized polyester (MP) film and low density poly ethylene (LDPE). Changes in lycopene content and non-enzymatic browning (NEB) were estimated during storage at room temperature. Pretreatment of 5 mm thickness of tomato slices with calcium chloride in combination with potassium metabisulphite and drying using a tunnel drier with subsequent storage of product in metalized polyester bags was selected as the best process.

Keywords: Drying pretreatments, Solar drying, Tomato powder, Tunnel drying

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Authors: Seung Pil Pack

Abstract:

Deaminated lesions were produced via nitrosative oxidation of natural nucleobases; uracul (Ura, U) from cytosine (Cyt, C), hypoxanthine (Hyp, H) from adenine (Ade, A), and xanthine (Xan, X) and oxanine (Oxa, O) from guanine (Gua, G). Such damaged nucleobases may induce mutagenic problems, so that much attentions and efforts have been poured on the revealing of their mechanisms in vivo or in vitro. In this study, we employed these deaminated lesions as useful probes for analysis of DNA-binding/recognizing proteins or enzymes. Since the pyrimidine lesions such as Hyp, Oxa and Xan are employed as analogues of guanine, their comparative uses are informative for analyzing the role of Gua in DNA sequence in DNA-protein interaction. Several DNA oligomers containing such Hyp, Oxa or Xan substituted for Gua were designed to reveal the molecular interaction between DNA and protein. From this approach, we have got useful information to understand the molecular mechanisms of the DNA-recognizing enzymes, which have not ever been observed using conventional DNA oligomer composed of just natural nucleobases.

Keywords: Deaminated lesion, DNA-protein interaction, DNA-recognizing enzymes

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Authors: Shu-Ying Marissa Pang, Stephen Tristram, Simon Brown

Abstract:

Candida spp. are common and aggressive pathogens. Because of the growing resistance of Candida spp. to current antifungals, novel targets, found in Candida spp. but not in humans or other flora, have to be identified. The alternative oxidase (AOX) is one such possibility. This enzyme is insensitive to cyanide, but is sensitive to compounds such as salicylhydroxamic acid (SHAM), disulfiram and n-alkyl gallates. The growth each of six Candida spp. was inhibited significantly by ~13 mM SHAM or 2 mM cyanide, albeit to differing extents. In C. dubliniensis, C. krusei and C. tropicalis the rate of O2 uptake was inhibited by 18-36% by 25 mM SHAM, but this had little or no effect on C. glabrata, C. guilliermondii or C. parapsilosis. Although SHAM substantially inhibited the growth of Candida spp., it is unlikely that the inhibition of AOX was the cause. Salicylhydroxamic acid is used therapeutically in the treatment of urinary tract infections and urolithiasis, but it also has some potential in the treatment of Candida spp. infection.

Keywords: alternative oxidase, Candida spp., growth, respiration, salicylhydroxamic acid.

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Authors: Khosro Mohammadi

Abstract:

Fertilization plays an important role in crop growth and soil improvement. This study was conducted to determine the best fertilization system for wheat production. Experiments were arranged in a complete block design with three replications in two years. Main plots consisted of six methods of fertilization including (N1): farmyard manure; (N2): compost; (N3): chemical fertilizers; (N4): farmyard manure + compost; (N5): farmyard manure + compost + chemical fertilizers and (N6): control were arranged in sub plots. The addition of compost or farm yard manure significantly increased the soil microbial biomass carbon in comparison to the chemical fertilizer. The dehydrogenase, phosphatase and urease activities in the N3 treatment were significantly lower than in the farm yard manure and compost treatments.

Keywords: Enzyme activity, fertilization, microbial biomasscarbon, wheat.

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Authors: Malinee Pongsavee

Abstract:

Sodium formate is the chemical substance used for food additive. Catalase is the important antioxidative enzyme in protecting the cell from oxidative damage by reactive oxygen species (ROS). The resultant level of oxidative stress in sodium formatetreated lymphocytes was investigated. The sodium formate concentrations of 0.05, 0.1, 0.2, 0.4 and 0.6 mg/mL were treated in human lymphocytes for 12 hours. After 12 treated hours, catalase activity change was measured in sodium formate-treated lymphocytes. The results showed that the sodium formate concentrations of 0.4 and 0.6 mg/mL significantly decreased catalase activities in lymphocytes (P < 0.05). The change of catalase activity in sodium formate-treated lymphocytes may be the oxidative damage marker for detect sodium formate exposure in human.

Keywords: Sodium formate, catalase activity, oxidative damage marker, toxicity.

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Authors: Asim Cengiz, Dede Basturk, Hakan Ozalp

Abstract:

Practicing recurring resistance workout such as may cause changes in human muscle. These changes may be because combination if several factors determining physical fitness. Thus, it is important to identify these changes. Several studies were reviewed to investigate these changes. As a result, the changes included positive modifications in amplified citrate synthase (CS) maximal activity, increased capacity for pyruvate oxidation, improvement on molecular signaling on human performance, amplified resting muscle glycogen and whole GLUT4 protein content, better health outcomes such as enhancement in cardiorespiratory fitness. Sprint training also have numerous long long-term changes inhuman body such as better enzyme action, changes in muscle fiber and oxidative ability. This is important because SV is the critical factor influencing maximal cardiac output and therefore oxygen delivery and maximal aerobic power.

Keywords: Sprint, training, performance, exercise.

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Authors: Ruchi Vyas, Sanjay Singh, Rashmi Sisodia

Abstract:

Chlorophytum borivillianum root extract (CBE) was chosen as a reducing agent to fabricate silver nanoparticles with the aim of studying its radioprotective efficacy. The formation of synthesized nanoparticles was characterized by UV–visible analysis (UV–vis), Fourier transform infra-red (FT-IR), Transmission electron microscopy (TEM), Scanning electron microscope (SEM). TEM analysis showed particles size in the range of 20-30 nm. For this study, Swiss albino mice were selected from inbred colony and were divided into 4 groups: group I- control (irradiated-6 Gy), group II- normal (vehicle treated), group III- plant extract alone and group IV- CB-AgNPs (dose of 50 mg/kg body wt./day) administered orally for 7 consecutive days before irradiation to serve as experimental. CB-AgNPs pretreatment rendered significant increase in body weight and testes weight at various post irradiation intervals in comparison to irradiated group. Supplementation of CB-AgNPs reversed the adverse effects of gamma radiation on biochemical parameters as it notably ameliorated the elevation in lipid peroxidation and decline in glutathione concentration in testes. These observations indicate the radio-protective potential of CB-AgNPs in testicular constituents against gamma irradiation in mice.

Keywords: Chlorophytum borivillianum, gamma radiation, radioprotective, silver nanoparticles.

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Authors: Supanun Kangrang, Kraipat Cheenkachorn, Kittiphong Rattanaporn, Malinee Sriariyanun

Abstract:

Rice straw is lignocellulosic biomass which can be utilized as substrate for the biogas production. However, due to the property and composition of rice straw, it is difficult to be degraded by hydrolysis enzymes. One of the pretreatment methods that modify such properties of lignocellulosic biomass is the application of lignocellulose-degrading microbial consortia. The aim of this study is to investigate the effect of microbial consortia to enhance biogas production. To select the high efficient consortium, cellulase enzymes were extracted and their activities were analyzed. The results suggested that microbial consortium culture obtained from cattle manure is the best candidate compared to decomposed wood and horse manure. A microbial consortium isolated from cattle manure was then mixed with anaerobic sludge and used as inoculum for biogas production. The optimal conditions for biogas production were investigated using response surface methodology (RSM). The tested parameters were the ratio of amount of microbial consortium isolated and amount of anaerobic sludge (MI:AS), substrate to inoculum ratio (S:I) and temperature. Here, the value of the regression coefficient R2 = 0.7661 could be explained by the model which is high to advocate the significance of the model. The highest cumulative biogas yield was 104.6 ml/g-rice straw at optimum ratio of MI:AS, ratio of S:I, and temperature of 2.5:1, 15:1 and 44°C respectively.

Keywords: Lignocellulolytic biomass, microbial consortium, cellulase, biogas, Response Surface Methodology.

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Authors: R. Kokilavani, K. Gurusamy, K. Arumugasamy

Abstract:

Protective effect of ethanolic extract of polyherbal formulation (PHF) was studied on carbon tetrachloride induced liver damage on carbon tetrachloride induced liver damage. Treatment of rats with 250mg /kg body weight of ethanolic extract of PHF protected rats against carbon tetrachloride liver injury by significant lowerering 5’ nucleotidase (5’NT), Gamma Glutamyl transferase (GGT), Glutamate dehdyrogenasse (GDH) and Succinate Dehydrogenase (SDH) levels compared to control. Normalization in these enzyme levels indicates strong hepatoprotective property of PHF extract.

Keywords: Carbon tetrachloride, ethanolic, hepatoprotective, polyherbal formulation.

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Authors: S. Gais, F. Fazouane, A. Mechakra

Abstract:

The present study describes the biosynthesis of a milkclotting protease by solid state fermentation (SSF) of a locally isolated mould, Rhizopus stolonifer. The production medium was prepared using wheat bran at 50% (w/v). The production conditions are optimized by varying 7 parameters: carbon and nitrogen sources, medium moisture, temperature, pH, fermentation time and inoculum-s size. The maximum enzyme synthesis was measured after 96 h of incubation time at temperature of 28°C. The optimum pH determined was 6 and the inoculum size was 3.106spores/ml. The optimum initial moisture content is comprised between 50 to 70%. The formation of milk clotting protease is enhanced when galactose and peptone are used at 10% (w/v) and 1% (w/v) concentrations respectively. The maximum production of milk clotting protease is 120 US/ml.

Keywords: Milk clotting activity, protease production, Rhizopus stolonifer, Solid state fermentation.

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Authors: M. S. Kilic, S. Korkut, B. Hazer

Abstract:

Newly synthesized Polypropylene-g-Polyethylene glycol polymer was first time used for a compartment-less enzymatic fuel cell. Working electrodes based on Polypropylene-g-Polyethylene glycol were operated as unmediated and mediated system (with ferrocene and gold/cobalt oxide nanoparticles). Glucose oxidase and bilirubin oxidase was selected as anodic and cathodic enzyme, respectively. Glucose was used as fuel in a single-compartment and membrane-less cell. Maximum power density was obtained as 0.65 nW cm-2, 65 nW cm-2 and 23500 nW cm-2 from the unmediated, ferrocene and gold/cobalt oxide modified polymeric film, respectively. Power density was calculated to be ~16000 nW cm-2 for undiluted wastewater sample with gold/cobalt oxide nanoparticles including system.

Keywords: Bilirubin oxidase, Enzymatic fuel cell, Glucose oxidase, Nanoparticles.

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Authors: S. M. Shahinul Islam, Israt Ara, Narendra Tuteja, Sreeramanan Subramaniam

Abstract:

Through anther and microspore culture methods, complete homozygous plants can be produced within a year as compared to the long inbreeding method. Isolated microspore culture is one of the most important techniques for rapid development of haploid plants. The efficiency of this method is influenced by several factors such as cultural conditions, growth regulators, plant media, pretreatments, physical and growth conditions of the donor plants, pollen isolation procedure, etc. The main purpose of this study was to improve the isolated microspore culture protocol in order to increase the efficiency of embryoids, its regeneration and reducing albinisms. Under this study we have tested mainly three different microspore isolation procedures by glass rod, homozeniger and by blending and found the efficiency on gametic embryogenesis. There are three types of media viz. washing, pre-culture and induction was used. The induction medium as AMC (modified MS) supplemented by 2, 4-D (2.5 mg/l), kinetin (0.5 mg/l) and higher amount of D-Manitol (90 g/l) instead of sucrose and two types of amino acids (L-glutamine and L-serine) were used. Out of three main microspore isolation procedure by homogenizer isolation (P₄) showed best performance on ELS induction (177%) and green plantlets (104%) compared with other techniques. For all cases albinisims occurred but microspore isolation from excised anthers by glass rod and homogenizer showed lesser numbers of albino plants that was also one of the important findings in this study.

Keywords: Androgenesis, pretreatment, microspore culture, regeneration, albino plants, Oryza sativa.

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Authors: Norlin Pauzi, Ahmad R.M. Yahya, Zairossani Nor, Amirul A. Abdullah

Abstract:

Strain M was isolated from the latex of Hevea brasiliensis that grow in the rubber farm area of Malaysia Rubber Board. Strain M was tentatively identified as Bacillus sp. Strain M demonstrated high protease production at pH 9, and this was suitable to be applied in rubber processing that was in alkaline conditions. The right and suitable proportion to be used in applying supernatant into the latex was two parts of latex and one part of enzyme. In this proportion, the latex was stable throughout the 72 hours of treatment. The potential of strain M to degrade protein in the natural rubber latex was proven with the reduction of 79.3% nitrogen in 24 hours treatment. Centrifugation process of the latex before undergoing the treatment had increased the protein degradation in latex. Although the centrifugation process did not achieve zero nitrogen content, it had improved the performance of protein denaturing in the natural rubber.

Keywords: Hevea brasiliensis, Bacillus sp., protease, latex.

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Authors: A. Nusrath Unissa, Sameer Hassan, Luke Elizabeth Hanna

Abstract:

Altered drug binding may be an important factor in isoniazid (INH) resistance, rather than major changes in the enzyme’s activity as a catalase or peroxidase (KatG). The identification of structural or functional defects in the mutant KatGs responsible for INH resistance remains as an area to be explored. In this connection, the differences in the binding affinity between wild-type (WT) and mutants of KatG were investigated, through the generation of three mutants of KatG, Ser315Thr [S315T], Ser315Asn [S315N], Ser315Arg [S315R] and a WT [S315]) with the help of software-MODELLER. The mutants were docked with INH using the software-GOLD. The affinity is lower for WT than mutant, suggesting the tight binding of INH with the mutant protein compared to WT type. These models provide the in silico evidence for the binding interaction of KatG with INH and implicate the basis for rationalization of INH resistance in naturally occurring KatG mutant strains of Mycobacterium tuberculosis.

Keywords: Mycobacterium tuberculosis, KatG, INH resistance, Mutants, Modeling, Docking.

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Authors: N. Matveev, A. Pervov

Abstract:

Local utilities often face problems of local industrial wastes, storm water disposal due to existing strict regulations. For many local industries, the problem of wastewater treatment and discharge into surface reservoirs can’t be solved through the use of conventional biological treatment techniques. Current discharge standards require very strict removal of a number of impurities such as ammonia, nitrates, phosphate, etc. To reach this level of removal, expensive reagents and sorbents are used. The modern concept of rational water resources management requires the development of new efficient techniques that provide wastewater treatment and reuse. As RO membranes simultaneously reject all dissolved impurities such as BOD, TDS, ammonia, phosphates etc., they become very attractive for the direct treatment of wastewater without biological stage. To treat wastewater, specially designed membrane "open channel" modules are used that do not possess "dead areas" that cause fouling or require pretreatment. A solution to RO concentrate disposal problem is presented that consists of reducing of initial wastewater volume by 100 times. Concentrate is withdrawn from membrane unit as sludge moisture. The efficient use of membrane RO techniques is connected with a salt balance in water system. Thus, to provide high ecological efficiency of developed techniques, all components of water supply and wastewater discharge systems should be accounted for.

Keywords: Reverse osmosis, stormwater treatment, openchannel module, wastewater reuse.

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Authors: Rashid S S, Alam M Z, Karim M I A, Salleh, M H

Abstract:

A statistical optimization was studied to design a media composition to produce optimum cellulolytic enzyme where palm oil mill effluent (POME) as a basal medium and filamentous fungus, Trichoderma reesei RUT-C30 were used in the liquid state bioconversion(LSB). 2% (w/v) total suspended solid, TSS, of the POME supplemented with 1% (w/v) cellulose, 0.5%(w/v) peptone and 0.02% (v/v) Tween 80 was estimated to produce the optimum CMCase activity of 18.53 U/ml through the statistical analysis followed by the faced centered central composite design(FCCCD). The probability values of cellulose (<0.0011) and peptone (0.0021) indicated the significant effect on the production of cellulase with the determination coefficient (R2) of 0.995.

Keywords: Face centered central composite design (FCCCD), Liquid state bioconversion (LSB), Palm oil mill effluent, Trichoderma reesei RUT C-30.

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Authors: N. N. A. K. Shah, R. A. Rahman, R. Shamsuddin, N. M. Adzahan

Abstract:

Pummelo is known to be the largest of all citrus fruits, with expected ratio of 2:1 (w/v) of producing juice, is an attractive opportunity for Malaysia to expand pummelo-s influence and marketability over the international market of juices. The purpose of this study is to identify and quantify the phenolic compounds in two Malaysian varieties of pummelo fruit juice: Ledang (PO55) and Tambun (PO52). Identifications of polyphenols composition were done using High Performance Liquid Chromatography Diode Array Detection (HPLC-DAD). The phenolic compounds that were found in both varieties were hydroxycinnamic acids and flavonones. This study proved that Tambun variety has the highest antioxidant and phenolic compounds in comparison to Ledang variety. However, considerations have to be made to suit consumer-s taste bud and the amount of enzyme needed to clarify the juice for its marketability.

Keywords: Antioxidant, HPLC, phenolic contents and pummelo fruit juice

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Authors: A.C. Duarte, C.I.L. Justino, K. Duarte, A.C. Freitas, R. Pereira, P. Chaves, P. Bettencourt, S. Cardoso, T.A.P. Rocha-Santos

Abstract:

This paper aims at overviewing the topics of a research project (CARDIOSENSOR) on the field of health sciences (biomaterials and biomedical engineering). The project has focused on the development of a nanosensor for the assessment of the risk of cardiovascular diseases by the monitoring of C-reactive protein (CRP), which has been currently considered as the best validated inflammatory biomarker associated to cardiovascular diseases. The project involves tasks such as: 1) the development of sensor devices based on field effect transistors (FET): assembly, optimization and validation; 2) application of sensors to the detection of CRP in standard solutions and comparison with enzyme-linked immunosorbent assay (ELISA); and 3) application of sensors to real samples such as blood and saliva and evaluation of their ability to predict the risk of cardiovascular disease.

Keywords: Carbon nanotubes field effect transistors, cardiovascular diseases, C-reactive protein, sensor.

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Authors: Yahia M., Laanani I., Benbia S., Hachemi M., Massinissa Y.

Abstract:

Uterine and oviducal fluids are necessary for capacitation of the spermatozoa and early embryonic development. The aim of the present study was to determine the effects of estrous cycle phases (follicular and luteal) on some biological parameters (enzymes, electrolytes and total proteins) in uterine and oviducal secretions of ewes. Oviducal and uterine fluids were collected, diluted and centrifuged. According to our results, concentrations of GPT, G6PDH, total proteins, K and Na were significantly (P <0.05) higher at the luteal phase, however, the levels of aldolase, Mg, Ca and P were significantly (P <0.05) higher at the follicular phase in uterine secretions. While, only oviducal K and Ca were significantly (P<0.05) higher at the follicular phase. Our study revealed the existence of significant cyclic variations for some uterine and oviducal parameters which indicates the effect of ovarian hormones on the components of genital secretions.

Keywords: Biochemical parameters, estrous cycle, ewe, genital secretion.

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Authors: I. Savitskaya, A. Kistaubayeva, A. Zhubanova, I. Blavachinskaiya, D. Ibrayeva, M. Abdulzhanova, A. Otarbay, A.Isabekova

Abstract:

This study was conducted for the investigation of number of cellulolytic bacteria and their ability in decomposition. Seven samples surface soil were collected on cellulose Zailiskii Alatau slopes. Cellulolitic activity of new strains of Bacillus, isolated from soil is determined. Isolated cellulose degrading bacteria were screened for determination of the highest cellulose activity by quantitative assay using Congo red, gravimetric assay and colorimetric DNS method trough of the determination of the parameters of sugar reduction. Strains are assigned to: B.subtilis, B.licheniformis, B. cereus and, В. megaterium. Bacillus strains consisting of several different types of cellulases have broad substrate specificity of cellulase complexes formed by them. Cellulolitic bacteria were recorded to have highest cellulase activity and selected for optimization of cellulase enzyme production.

Keywords: Cellulose-degrading bacteria, cellulase complex, foothills soil, screening.

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Authors: Ahmad Manbohi, Seyyed Hamid Ahmadi

Abstract:

A low-cost paper-based microfluidic device (PAD) for the multiplex electrochemical determination of glucose, uric acid, and dopamine in biological fluids was developed. Using wax printing, PAD containing a central zone, six channels, and six detection zones was fabricated, and the electrodes were printed on detection zones using pre-made electrodes template. For each analyte, two detection zones were used. The carbon working electrode was coated with chitosan-BSA (and enzymes for glucose and uric acid). To detect glucose and uric acid, enzymatic reactions were employed. These reactions involve enzyme-catalyzed redox reactions of the analytes and produce free electrons for electrochemical measurement. Calibration curves were linear (R^² > 0.980) in the range of 0-80 mM for glucose, 0.09–0.9 mM for dopamine, and 0–50 mM for uric acid, respectively. Blood samples were successfully analyzed by the proposed method.

Keywords: Multiplex, microfluidic paper-based electrochemical biosensors, biomarkers, biological fluids.

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Authors: Jayashree Ramana

Abstract:

An alarming emergence of multidrug-resistant strains of the tuberculosis pathogen Mycobacterium tuberculosis and continuing high worldwide incidence of tuberculosis has invigorated the search for novel drug targets. The enzyme glutamate racemase (MurI) in bacteria catalyzes the stereoconversion of L-glutamate to D-glutamate which is a component of the peptidoglycan cell wall of the bacterium. The inhibitors targeted against MurI from several bacterial species have been patented and are advocated as promising antibacterial agents. However there are none available against MurI from Mycobacterium tuberculosis, due to the lack of its threedimensional structure. This work accomplished two major objectives. First, the tertiary structure of MtMurI was deduced computationally through homology modeling using the templates from bacterial homologues. It is speculated that like in other Gram-positive bacteria, MtMurI exists as a dimer and many of the protein interactions at the dimer interface are also conserved. Second, potent candidate inhibitors against MtMurI were identified through docking against already known inhibitors in other organisms.

Keywords: Glutamate racemase, homology modeling, docking, drug resistance.

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Authors: Sirilak Namwong , Wonnop Visessanguan, Soottawat Benjakul, Somboon Tanasupawat

Abstract:

The halophilic proteinase showed a maximal activity at 50°C and pH 9~10, in 20% NaCl and was highly stabilized by NaCl. It was able to hydrolyse natural actomyosin (NAM), collagen and anchovy protein. For NAM hydrolysis, the myosin heavy chain was completely digested by halophilic proteinase as evidenced by the lowest band intensity remaining, but partially hydrolysed actin. The SR5-3 proteinase was also capable hydrolyzing two major components of collagen, β- and α-compounds, effectively. The degree of hydrolysis (DH) of the halophilic proteinase and commercial proteinases (Novozyme, Neutrase, chymotrypsin and Flavourzyme) on the anchovy protein, were compared, and it was found that the proteinase showed a greater degree of hydrolysis towards anchovy protein than that from commercial proteinases. DH of halophilic proteinase was sharply enhanced according to the increase in the concentration of enzyme from 0.035 U to 0.105 U. The results warranting that the acceleration of the production of fish sauce with higher quality, may be achieved by adding of the halophilic proteinase from this bacterium.

Keywords: Halophilic proteinase, Halobacillus sp. SR5-3, anchovy (Spolephorus spp.) proteins, fish sauce

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Authors: D.T. Mirzarakhmetova

Abstract:

The studying of enzymatic esterification of carboxylic acids and higher alcohols was performed by esterase Saccharomyces cerevisiae in water-organic medium. Investigation of the enzyme specificity to acetic substrates showed the best result with acetic acid in esterification reactions with ethanol whereas within other carboxylic acids the esterification decreased with acids: hexanoic > pentanoic > butyric > decanoic. In relation to higher alcohols C3-C5, esterification increased with alcohols propanol < butanol < amylol. Also it was determined that esterase was more specific to alcohols with branched chain such as isobutyl alcohol and isoamyl alcohol. Data obtained may have important practical implications, for example, for application of yeast esterase in producing various volatile esters as well as in enzymatic transformation of volatile acids and toxic fusel alcohols into volatile esters by providing the production of the high quality alcoholic beverages with redused content of higher alcohols as well as with improved degustational and hygienic properties.

Keywords: enzymes in non-conventional media, esterification, higher alcohols, volatile esters, yeast esterase

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Authors: Vedansh Gupta, Allyson Lutz, Ameen Razavi, Fatemeh Shirazi

Abstract:

The agriculture industry is especially vulnerable to forecasted water shortages. In the fresh and fresh-cut produce sector, conventional flume-based washing with recirculation exhibits high water demand. This leads to a large water footprint and possible cross-contamination of pathogens. These can be alleviated through advanced water reuse processes, such as membrane technologies including reverse osmosis (RO). Water reuse technologies effectively remove dissolved constituents but can easily foul without pre-treatment. Biological treatment is effective for the removal of organic compounds responsible for fouling, but not at the low temperatures encountered at most produce processing facilities. This study showed that the Microvi MicroNiche Engineering (MNE) technology effectively removes organic compounds (> 80%) at low temperatures (6-8 °C) from wash water. The MNE technology uses synthetic microorganism-material composites with negligible solids production, making it advantageously situated as an effective bio-pretreatment for RO. A preliminary technoeconomic analysis showed 60-80% savings in operation and maintenance costs (OPEX) when using the Microvi MNE technology for organics removal. This study and the accompanying economic analysis indicated that the proposed technology process will substantially reduce the cost barrier for adopting water reuse practices, thereby contributing to increased food safety and furthering sustainable water reuse processes across the agricultural industry.

Keywords: Biological pre-treatment, innovative technology, vegetable processing, water reuse, agriculture, reverse osmosis, MNE biocatalysts.

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Authors: Hazim Saadoon Aljewari, Mohammed Ibraheem Nader, Abdul Hussain M. Alfaisal, NatthidaWeerapreeyakul, Sahapat

Abstract:

L-asparaginase was extracted from pathogenic Escherichia coli which was isolated from urinary tract infection patients. L-asparaginase was purified 96-fold by ultrafiltration, ion exchange and gel filtration giving 39.19% yield with final specific activity of 178.57 IU/mg. L-asparaginase showed 138,356±1,000 Dalton molecular weight with 31024±100 Dalton molecular mass. Kinetic properties of enzyme resulting 1.25×10-5 mM Km and 2.5×10-3 M/min Vmax. L-asparaginase showed a maximum activity at pH 7.5 when incubated at 37 ºC for 30 min and illustrated its full activity (100%) after 15 min incubation at 20-37 ºC, while 70% of its activity was lost when incubated at 60 ºC. L-asparaginase showed cytotoxicity to U937 cell line with IC50 0.5±0.19 IU/ml, and selectivity index (SI=7.6) about 8 time higher selectivity over the lymphocyte cells. Therefore, the local pathogenic E. coli strains may be used as a source of high yield of L-asparaginase to produce anti cancer agent with high selectivity.

Keywords: L-asparaginase, Purification, Cytotoxicity, selectivity index

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