Search results for: industrial enzymes production

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

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Extracellular laccases are copper-containing microbial enzymes with many industrial biotechnological applications. This study evaluated the ability of nutrients in coconut coir to enhance the yield of extracellular laccase of Galactomyces reesii IFO 10823 and develop a co-culture between this yeast and other filamentous fungi isolated from the fungus comb of Macrotermes sp. The co-culture between G. reesii IFO 10823 and M. indicus FJ-M-5 (G3) gave the highest activity at 580.20 U/mL. When grown in fermentation media prepared from coconut coir and distilled water at 70% of initial moisture without supplement addition, G3 produced extracellular laccase of 113.99 U/mL.

Keywords: extracellular laccase, production, yeast, natural inducer

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Authors: H. M. Takematsu, B. R. De Camargo, E. F. Noronha

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The use of hydrolyzing enzymes for degradation of lignocellulosic biomass is of great concern for the production of second generation ethanol. Although many hollocelulases have already been described in the literature, much more has to be discovered. Therefore, the aim of this study to evaluate hollocelulase production of P. polonicum grown in liquid media containing sugarcane bagasse as the carbon source. From a collection of twenty fungi isolated from Cerrado biome soil, P. polonicum was molecular identified by sequencing of ITS4, βtubulin and Calmodulin genes, and has been chosen to be further investigated regarding its potential production of hydrolyzing enzymes. Spore suspension (1x10-6 ml-1) solution was inoculated in sterilized minimal liquid medium containing 0,5%(w/v) of non-pretreated sugarcane bagasse as the carbon source, and incubated in shaker incubator at 28°C and 120 rpm. The supernatant obtained, was subjected to enzymatic assays to analyze xylanase, mannanase, pectinase and endoglucanase activities. Xylanase activity showed better results (67,36 UI/mg). Xylanases bands were indicated by zymogram and SDS-PAGE, and one of them was partially purified and characterized. It showed maximum activity at 50 °C, was thermostable for 72h at 40°C, and pH5.0 was the optimum observed. This study presents P. polonicum as an interesting source of hollocelulases, especially xylanase, for lignocellulose bio-conversion processes with commercial use.

Keywords: sugarcane bagasse, Cerrado biome , hollocelulase, lignocellulosic biomass

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Authors: M. Umar Dahot, G. S. Mangrio

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In this study Agro-industrial waste was used as a carbon source, which is a low cost substrate. Along with this, various sugars and molasses of 2.5% and 5% were investigated as substrate/carbon source for the growth of A.niger and Pectinase production. Different nitrogen sources were also used. An overview of results obtained show that 5% sucrose, 5% molasses and 0.4% (NH4)2SO4 were found the best carbon and nitrogen sources for the production of pectinase by A. niger. The maximum production of pectinase (26.87units/ml) was observed at pH 6.0 after 72 hrs incubation. The optimum temperature for the maximum production of pectinase was achieved at 35ºC when maximum production of pectinase was obtained as 28.25Units/ml.Pectinase enzyme was purified with ammonium sulphate precipitation and dialyzed sample was finally applied on gel filtration chromatography (Sephadex G-100) and Ion Exchange DEAE A-50. The enzyme was purified 2.5 fold by gel chromatography on Sephadex G-100 and Four fractions were obtained, Fraction 1, 2, 4 showed single band while Fraction -3 showed multiple bands on SDS Page electrophoresis. Fraction -3 was pooled, dialyzed and separated on Sephdex A-50 and two fractions 3a and 3b showed single band. The molecular weights of the purified fractions were detected in the range of 33000 ± 2000 and 38000± 2000 Daltons. The purified enzyme was specifically most active with pure pectin, while pectin, Lemon pectin and orange peel given lower activity as compared to (control). The optimum pH and temperature for pectinase activity was found between pH 5.0 and 6.0 and 40°- 50°C, respectively. The enzyme was stable over the pH range 3.0-8.0. The thermostability of was determined and it was observed that the pectinase activity is heat stable and retains activity more than 40% when incubated at 90°C for 10 minutes. The pectinase activity of F3a and F3b was increased with different metal ions. The Pectinase activity was stimulated in the presence of CaCl2 up to 10-30%. ZnSO4, MnSO4 and Mg SO4 showed higher activity in fractions F3a and F3b, which indicates that the pectinase belongs to metalo-enzymes. It is concluded that A. niger is capable to produce pH stable and thermostable pectinase, which can be used for industrial purposes.

Keywords: pectinase, a. niger, production, purification, characterization

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Authors: Jessica Pinheiro Silva, Brenda Rabello De Camargo, Daniel Gusmao De Morais, Eliane Ferreira Noronha

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The utilization of lignocellulosic biomass as source of polysaccharides for industrial applications requires an arsenal of enzymes with different mode of action able to hydrolyze its complex and recalcitrant structure. Clostridium thermocellum is gram-positive, thermophilic bacterium producing lignocellulosic hydrolyzing enzymes in the form of multi-enzyme complex, termed celulossomes. This complex has several hydrolytic enzymes attached to a large and enzymically inactive protein known as Cellulosome-integrating protein (CipA), which serves as a scaffolding protein for the complex produced. This attachment occurs through specific interactions between cohesin modules of CipA and dockerin modules in enzymes. The present work aims to construct celulosomes in vitro with the structural protein CipA, a xylanase called Xyn10D and a cellulose called CelJ from C.thermocellum. A mini-scafoldin was constructed from modules derived from CipA containing two cohesion modules. This was cloned and expressed in Escherichia coli. The other two genes were cloned under the control of the alcohol oxidase 1 promoter (AOX1) in the vector pPIC9 and integrated into the genome of the methylotrophic yeast Pichia pastoris GS115. Purification of each protein is being carried out. Further studies regarding enzymatic activity of the cellulosome is going to be evaluated. The cellulosome built in vitro and composed of mini-CipA, CelJ and Xyn10D, can be very interesting for application in industrial processes involving the degradation of plant biomass.

Keywords: cellulosome, CipA, Clostridium thermocellum, cohesin, dockerin, yeast

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Authors: Hakan Arslan, Deniz Ekinci, Alper Gungor, Gurkan Bilir, Omer Tas, Mehmet Altun

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Drought is one of the major abiotic stresses affecting the agricultural production worldwide. In this study, Leaf samples were taken from the carrot plants grown under drought stress conditions during the harvesting period. The plants were irrigated in three irrigation interval (4, 6 and 8 days) and Irrigation water regime was set up in pots. The changes in activities of antioxidant enzymes such as glutathione reductase (GR), glutathione s-transferase (GST), superoxide dismutase (SOD)) in leaves of black carrot were investigated. The activities of antioxidant enzymes (GR, GST, SOD) were varied significantly with irrigation intervals. The highest value of GR, GST and SOD were determined in the irrigation interval of 6 days. All antioxidant activity values were decreased in 8 days of irrigation interval. As a result of the study, it has been suggested that optimum irrigation intervals for plants can be used in antioxidant enzymes.

Keywords: antioxidant enzyme, carrot, drought, irrigation interval

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Authors: P. Pasomboon, P. Chumnanpuen, T. E-kobon

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Hyaluronic acid (HA) consists of linear heteropolysaccharides repeat of D-glucuronic acid and N-acetyl-D-glucosamine. HA has various useful properties to maintain skin elasticity and moisture, reduce inflammation, and lubricate the movement of various body parts without causing immunogenic allergy. HA can be found in several animal tissues as well as in the capsule component of some bacteria including Pasteurella multocida. This study aimed to modify a genome-scale metabolic model of Escherichia coli using computational simulation and flux analysis methods to predict HA productivity under different carbon sources and nitrogen supplement by the addition of two enzymes (GLMU2 and HYAD) from P. multocida to improve the HA production under the specified amount of carbon sources and nitrogen supplements. Result revealed that threonine and aspartate supplement raised the HA production by 12.186%. Our analyses proposed the genome-scale metabolic model is useful for improving the HA production and narrows the number of conditions to be tested further.

Keywords: Pasteurella multocida, Escherichia coli, hyaluronic acid, genome-scale metabolic model, bioinformatics

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Authors: Fiyinfoluwa Giwa, Nicholas Ngepah

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This paper investigates the relationship between the Fourth Industrial Revolution and food security in South Africa. The Ordinary Least Square was adopted from 2012 Q1 to 2021 Q4. The study used artificial intelligence investment and the food production index as the measure for the fourth industrial revolution and food security, respectively. Findings reveal a significant and positive coefficient of 0.2887, signifying a robust statistical relationship between AI adoption and the food production index. As a policy recommendation, this paper recommends the introduction of incentives for farmers and agricultural enterprises to adopt AI technologies -and the expansion of digital connectivity and access to technology in rural areas.

Keywords: Fourth Industrial Revolution, food security, artificial intelligence investment, food production index, ordinary least square

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Authors: Joseph A. Bentil, Anders Thygesen, Lene Langea, Moses Mensah, Anne Meyer

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The study investigated the saccharification potential of in-house enzymes produced from a white-rot basidiomycete strain, Polyporus ciliatus CBS 366.74. The in-house enzymes were produced by growing the fungus on mono and composite substrates of cocoa pod husk (CPH) and green seaweed (GS) (Ulva lactuca sp.) with and without the addition of 25mM ammonium nitrate at 4%w/v substrate concentration in submerged condition for 144 hours. The crude enzyme extracts preparations (CEE 1-5 and CEE 1-5+AN) obtained from the fungal cultivation process were sterile-filtered and used as enzyme sources for enzymatic hydrolysis of hydrothermally pretreated corn stover using a commercial cocktail enzyme, Cellic Ctec3, as benchmark. The hydrolysis was conducted at 50ᵒC with 50mM sodium acetate buffer, pH 5 based on enzyme dosages of 5 and 10 CMCase Units/g biomass at 1%w/v dry weight substrate concentration at time points of 6, 24, and 72 hours. The enzyme activity profile of the in-house enzymes varied among the growth substrates with the composite substrates (50-75% GS and AN inclusion), yielding better enzyme activities, especially endoglucanases (0.4-0.5U/mL), β-glucosidases (0.1-0.2 U/mL), and xylanases (3-10 U/mL). However, nitrogen supplementation had no significant effect on enzyme activities of crude extracts from 100% GS substituted substrates. From the enzymatic hydrolysis, it was observed that the in-house enzymes were capable of hydrolysing the pretreated corn stover at varying degrees; however, the saccharification yield was less than 10%. Consequently, theoretical glucose yield was ten times lower than Cellic Ctec3 at both dosage levels. There was no linear correlation between glucose yield and enzyme dosage for the in-house enzymes, unlike the benchmark enzyme. It is therefore recommended that the in-house enzymes are used to complement the dosage of commercial enzymes to reduce the cost of biomass saccharification.

Keywords: enzyme production, hydrolysis yield, feedstock, enzyme blend, Polyporus ciliatus

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Authors: Mohamed Elbehery

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Historical textiles in the burial environment or in museums are exposed to many types of stains and dirt that are associated with historical textiles by multiple chemical bonds that cause damage to historical textiles. The cleaning process must be carried out with great care, with no irreversible damage, and sediments removed without affecting the original material of the surface being cleaned. Science and technology continue to provide innovative systems in the bio-cleaning process (using pure enzymes) of historical textiles and artistic surfaces. Lipase and α-amylase were immobilized on nanoparticles of alginate/κ-carrageenan nanoparticle complex and used in historical textiles cleaning. Preparation of nanoparticles, activation, and enzymes immobilization were characterized. Optimization of loading time and units of the two enzymes were done. It was found that, the optimum time and units of amylase were 4 hrs and 25U, respectively. While, the optimum time and units of lipase were 3 hrs and 15U, respectively. The methods used to examine the fibers using a scanning electron microscope equipped with an X-ray energy dispersal unit: SEM with EDX unit.

Keywords: nanoparticles, enzymes, immobilization, textiles

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Authors: S. Ameddah, O. Deffa, H. Aissaoui, A. Menad, R. Mekkiou, F. Benayache, S. Benayache

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Acetylhydrazide (ACHD) is a metabolite of the anti-tubercular drug isoniazid (INH) that has been implicated in liver damage. This study was designed to evaluate hapatoprotective of n-BuOH extract of Heliotrpium undulatum (HUBE) in ACHD hepatotoxicity in rats. Hepatic damage was induced by administration of ACHD (300 mg/Kg op). The protection was affected by the administration of HUBE (200 mg/Kg op) for 14 days before ACHD administration, caused a decrease in LPO levels and in the transaminase and ALP levels and restored the GSH and its related enzymes (GPx, GST, GR) (50-62 %). Simultaneous administration of HUBE afforded a partial protection in statue of hepatic GSH conjugating enzymes upon administration of ACHD.

Keywords: heliotrpium undulatum, acetylhydrazide, glutathione conjugating enzymes, oxydatif stress, heaptoprotectif effect

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

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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 method that modifies 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 (RSM)

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Authors: Nadjib Melkemi, Salah Belaidi

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Quantitative Structure-Activity Relationship (QSAR) studies have been performed on nineteen molecules of 1,2-dithiole-3-thione analogues. The compounds used are the potent inducers of enzymes involved in the maintenance of reduced glutathione pools as well as phase-2 enzymes important to electrophile detoxication. A multiple linear regression (MLR) procedure was used to design the relationships between molecular descriptor and detoxication properties of the 1,2-dithiole-3-thione derivatives. The predictivity of the model was estimated by cross-validation with the leave-one-out method. Our results suggest a QSAR model based of the following descriptors: qS2, qC3, qC5, qS6, DM, Pol, log P, MV, SAG, HE and EHOMO for the specific activity of quinone reductase; qS1, qS2, qC3, qC4, qC5, qS6, DM, Pol, logP, MV, SAG, HE and EHOMO for the production of growth hormone. To confirm the predictive power of the models, an external set of molecules was used. High correlation between experimental and predicted activity values was observed, indicating the validation and the good quality of the derived QSAR models.

Keywords: QSAR, quinone reductase activity, production of growth hormone, MLR

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Authors: A. U. Adamu, Hamisu Abdu, A. A. Saidu

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The enzymatic synthesis of 3-O-β-D-glucopyranoside-betulinic acid using Novozyme-435 as a catalyst was studied. The effect of various parameters such as substrate molar ratio, reaction temperature, reaction time, re-used enzymes and amount of enzymes were investigated. The optimum rection conditions for the enzymatic glycosidation of betulinic acid in an organic solvent using Novozym-435 was found to be at 1:1.2 substrate molar ratio, 55oC, 24 h and 180 mg of enzymes with percentage conversion of 88.69 %.

Keywords: betulinic acid, glycosidation, novozyme-435, optimization

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Authors: Vandana Bali, Manab B. Bera, Parmjit S. Panesar

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Microbial production of antimicrobials as biopreservatives is the major area of focus nowadays due to increased interest of consumers towards natural and safe preservation of ready to eat food products. The agro-industrial byproduct based medium and optimized process conditions can contribute in economical production of bacteriocins. Keeping this in view, the present investigation was carried out on agro-industrial byproducts utilization for the production of bacteriocin using Enterococcus faecium BS13 isolated from local fermented food. Different agro-industrial byproduct based carbon sources (whey, potato starch liquor, kinnow peel, deoiledrice bran and molasses), nitrogen sources (soya okra, pea pod and corn steep liquor), metal ions and surfactants were tested for optimal bacteriocin production. The effect of various process parameters such as pH, temperature, inoculum level, agitation and time were also tested on bacteriocin production. The optimized medium containing whey, supplemented with 4%corn steep liquor and polysorbate-80 displayed maximum bacteriocin activity with 2% inoculum, at pH 6.5, temperature 40oC under shaking conditions (100 rpm).

Keywords: Bacteriocin, biopreservation, corn steep liquor, Enterococcus faecium, waste utilization, whey

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Authors: Mehmet Savsar

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This paper presents a practical case application of simulation modeling and analysis in a specific industrial setting. Various maintenance related parameters of the equipment in the system under consideration are determined and a simulation model is developed to study system behavior. System performance is determined based on established parameters and operational policies, which included system operation with and without preventive maintenance implementation. The results show that preventive maintenance practice has significant effects on improving system productivity. The simulation procedures outlined in this paper can be used by operation managers to perform production line analysis under different maintenance policies in various industrial settings.

Keywords: simulation, production line, machine failures, maintenance, industrial bakery

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Authors: L. Heidari, M. Jalili Ghazizade

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In recent years, different petrochemical complexes have been established to produce aromatic compounds. Among them, Nouri Petrochemical Complex (NPC) is the largest producer of aromatic raw materials in the world, and is located in south of Iran. Environmental concerns have been raised in this region due to generation of different types of solid waste generated in the process of aromatics production, and subsequently, industrial waste characterization has been thoroughly considered. The aim of this study is qualitative and quantitative characterization of industrial waste generated in the aromatics production process and determination of the best method for industrial waste management. For this purpose, all generated industrial waste during the production process was determined using a checklist. Four main industrial wastes were identified as follows: spent industrial soil, spent catalyst, spent molecular sieves and spent N-formyl morpholine (NFM) solvent. The amount of heavy metals and organic compounds in these wastes were further measured in order to identify the nature and toxicity of such a dangerous compound. Then industrial wastes were classified based on lab analysis results as well as using different international lists of hazardous waste identification such as EPA, UNEP and Basel Convention. Finally, the best method of waste disposal is selected based on environmental, economic and technical aspects. 

Keywords: aromatic compounds, industrial soil, molecular sieve, normal formyl morpholine solvent

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Authors: Bassam Al Johny

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Halophilic bacteria are organisms which thrive in salt-rich environments, such as salt lakes, solar salterns and salt mines which contain large populations of these organisms. In biotechnology, such salt-tolerant bacteria are widely used for the production of valuable enzymes, and more than a thousand years ago humans began using salt to cure and thereby preserve perishable foods and other materials, such as hides; halophiles can be detrimental to the preservation of salt brine cured hides. The aim of this review is to provide an overview of the taxonomy of these organisms including novel isolates from rock salt, and also to discuss their current and future biotechnological and environmental uses.

Keywords: hypersaline environments, halophilic bacteria, environmental application, industrial application

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Authors: Sunbul Jassim Hamodi, Muna Khalid Khudayer, Firas Muzahem Hussein

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The experiment was conducted to study the effect of supplement sources of Phytase enzyme (bacterial, fungal, enzymes mixture) using levels (250, 500, 750) FTY/ kg feed to diets compared with control on the performance for one thousand fifty broiler chicks (Ross 308) from 1day old with initial weight 39.78 gm till 42 days. The study involved 10 treatments, three replicates per treatment (35 chicks/replicate). Treatments were as follows: T1: control diet (without any addition). T2: added bacterial phytase enzyme 250FTY/ kg feed. T3: added bacterial phytase enzyme 500FTY/ kg feed. T4: added bacterial phytase enzyme 750FTY/ kg feed. T5: added fungal phytase enzyme 250FTY/ kg feed. T6: added fungal phytase enzyme 500FTY/ kg feed. T7: added fungal phytase enzyme 750FTY/ kg feed. T8 added enzymes mixture 250U/ kg feed. T9: added enzymes mixture 500U/ kg feed. T10: added enzymes mixture 750U/ kg feed. The results revealed that supplementing 750 U from enzymes mixture to broiler diet increased significantly (p <0.05) body weight compared with (250 FTY bacterial phytase/Kgfeed), (750 FTY bacterial phytase/Kg feed), (750FTY fungal phytase/Kgfeed) at 6 weeks, also supplemented different sources and levels from phytase enzyme improved a cumulative weight gain for (500 FTY bacterial phytase/Kgfeed), (250FTY fungal phytase/Kgfeed), (500FTY fungal phytase/Kgfeed), (250 Uenzymes mixture/Kgfeed), (500 Uenzymes mixture/Kgfeed) and (750 U enzymes mixture/Kgfeed) treatments compared with (750 FTY fungal phytase/Kgfeed)treatment, about accumulative feed consumption (500 FTY fungal phytase/Kgfeed) and (250 Uenzymes mixture/Kgfeed) increased significantly compared with control group and (750FTY fungal phytase/Kgfeed) during 1-6 weeks. There were significantly improved in cumulative feed conversion for (500U enzymes mixture/Kgfeed) compared with the worse feed conversion ratio that recorded in (250 FTY bacterial phytase/Kgfeed). No significant differences between treatments in internal organs relative weights, carcass cuts, dressing percentage and production index. Mortality was increased in (750FTY fungal phytase/Kgfeed) compared with other treatments.

Keywords: phytase, phytic acid, broiler, productive performance

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Authors: José Luiz da Silva Neto, Renato Sipelli Silva, Érick Aragão Ribeiro

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The automation of industrial processes plays a vital role in industries today, becoming an integral and important part of the industrial process and modern production. The process control systems are designed to maximize production, reduce costs and minimize risks in production. However, these systems are generally not deployed methodologies and planning. So that this article describes the development of an automation system of a kitchen preparation of chemicals in the textile industry based on a retrofitting methodology that provides more quality into the process at a lower cost.

Keywords: automation, textile industry, kitchen chemical, information integration

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Authors: Natcha Ruamyat, Nichakorn Khondee

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The potential of an alkaliphilic bacteria isolated from soil in Thailand to utilized agro-industrial and agricultural wastes for the production of biosurfactants was evaluated in this study. Among five isolates, Pseudomonas mendocina NK41 used soapstock as substrate showing a high biosurfactant concentration of 7.10 g/L, oil displacement of 97.8 %, and surface tension reduction to 29.45 mN/m. Various agricultural residues were applied as mixed substrates with soapstock to enhance the synthesis of biosurfactants. The production of biosurfactant and bacterial growth was found to be the highest with coconut oil cake as compared to Sacha inchi shell, coconut kernel cake, and durian shell. The biodegradability of agro-industrial wastes was better than agricultural wastes, which allowed higher bacterial growth. The pretreatment of coconut oil cake by combined alkaline and hydrothermal method increased the production of biosurfactant from 12.69 g/L to 13.82 g/L. The higher microbial accessibility was improved by the swelling of the alkali-hydrothermal pretreated coconut oil cake, which enhanced its porosity and surface area. The pretreated coconut oil cake was reused twice in the repeated batch production, showing higher biosurfactant concentration up to 16.94 g/L from the second cycle. These results demonstrated the capability of using lignocellulosic wastes from agricultural and agro-industrial activities to produce a highly valuable biosurfactant. High biosurfactant yield with low-cost substrate reveals its potential towards further commercialization of biosurfactant on large-scale production.

Keywords: alkaliphilic bacteria, agricultural/agro-industrial wastes, biosurfactant, combined alkaline-hydrothermal pretreatment

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Authors: L. Kutateldze, T. Urushadze, R. Khvedelidze, N. Zakariashvili, I. Khokhashvili, T. Sadunishvili

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Microbial cellulase/xylanase have shown their potential application in various industries including pulp and paper, textile, laundry, biofuel production, food and feed industry, brewing, and agriculture. Extremophilic micromycetes and their enzymes that are resistant to critical values of temperature and pH, and retaining enzyme activity for a long time are of great industrial interest. Among strains of microscopic fungi from the collection of S. Durmishidze Institute of Biochemistry and Biotechnology, strains isolated from different ecological niches of Southern Caucasus-active producers of cellulase/xylanase have been selected by means of screening under deep cultivation conditions. Extremophilic micromycetes and their enzymes that are resistant to critical values of temperature and pH, and retaining enzyme activity for a long time are of great industrial interest. Among strains of microscopic fungi from the collection of S. Durmishidze Institute of Biochemistry and Biotechnology, strains isolated from different ecological niches of Southern Caucasus-active producers of cellulase/xylanase have been selected by means of screening under deep cultivation conditions. Representatives of the genera Aspergillus, Penicillium and Trichoderma are outstanding by relatively high activities of these enzymes. Among the producers were revealed thermophilic strains, representatives of the genus Aspergillus-Aspergillus terreus, Aspergillus versicolor, Aspergillus wentii, also strains of Sporotrichum pulverulentum and Chaetomium thermophile. As a result of optimization of cultivation media and conditions, activities of enzymes produced by the strains have been increased by 4 -189 %. Two strains, active producers of cellulase/xylanase – Penicillium canescence E2 (mesophile) and Aspergillus versicolor Z17 (thermophile) were chosen for further studies. Cellulase/xylanase enzyme preparations from two different genera of microscopic fungi Penicillium canescence E2 and Aspergillus versicolor Z 17 were obtained with activities 220 U/g /1200 U/g and 125 U/g /940 U/g, correspondingly. Main technical characteristics were as follows: the highest enzyme activities were obtained for mesophilic strain Penicillium canescence E2 at 45-500C, while almost the same enzyme activities were fixed for the thermophilic strain Aspergillus versicolor Z 17 at temperature 60-65°C, exceeding the temperature optimum of the mesophile by 150C. Optimum pH of action of the studied cellulase/xylanases from mesophileic and thermophilic strains were similar and equaled to 4.5-5.0 It has been shown that cellulase/xylanase technical preparations from selected strains of Penicillium canescence E2 and Aspergillus versicolor Z17 hydrolyzed cellulose of untreated wheat straw to reducible sugars by 46-52%, and to glucose by 22-27%. However the thermophilic enzyme preparations from the thermophilic A.versicolor strains conducted the process at 600C higher by 100C as compared to mesophlic analogue. Rate of hydrolyses of the pretreated substrate by the same enzyme preparations to reducible sugars and glucose conducted at optimum for their action 60 and 500C was 52-61% and 29-33%, correspondingly. Thus, maximum yield of glucose and reducible sugars form untreated and pretreated wheat straw was achieved at higher temperature (600C) by enzyme preparations from thermophilic strain, which gives advantage for their industrial application.

Keywords: cellulase/xylanase, cellulose hydrolysis, microscopic fungi, thermophilic strain

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Authors: Reshmi Vijayan

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Alkaline protease is one of the most important enzymes in the biological world. Microbial production of alkaline protease is getting more attention from researchers due to its unique properties and substantial activity. Microorganisms are the most common sources of commercial enzymes due to their physiological and biochemical properties. The study was conducted on Ayiramthenghu mangrove sediments to isolate protease producing bacteria. All the isolates were screened for proteolytic activity on a skim milk agar plate at 37˚C for 48hrs. Protease activities were determined by the formation of a clear zone around the colonies on Skim milk agar medium. The activity of the enzyme was measured by the tyrosine standard curve, and it was found to be 0.186285 U/ml/min.

Keywords: protease, protease assay, skim milk agar medium, mangrove ecosystem

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Authors: Martina Perinkova, Lenka Kolarcikova, Marketa Twrda

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Brownfields are one of the most important problems that must be solved by today's cities. The topic of this article is description of developing a comprehensive transformation of post-industrial area of the former iron factory national cultural heritage Lower Vítkovice. City of Ostrava used to be industrial superpower of the Czechoslovak Republic, especially in the area of coal mining and iron production, after declining industrial production and mining in the 80s left many unused areas of former factories generally brownfields and backfields. Since the late 90s we are observing how the city officials or private entities seeking to remedy this situation. Regeneration of brownfields is a very expensive and long-term process. The area is now rebuilt for tourists and residents of the city in the entertainment, cultural, and social center. It was necessary do the reconstruction of the industrial monuments. Equally important was the construction of new buildings, which helped reusing of the entire complex. This is a unique example of transformation of technical monuments and completion of necessary new objects, so that the area could start working again and reintegrate back into the urban system.

Keywords: brown fields, conversion, historical and industrial buildings, reconstruction

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Authors: Justina I. R. Udotong, Essien U. Essien

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Diagnostic enzymes like aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) were determined as indices of heavy metal pollution in Tilapia guinensis. Three different sets of fishes treated with lead (Pb), iron (Fe) and copper (Cu) were used for the study while a fourth group with no heavy metal served as a control. Fishes in each of the groups were exposed to 2.65 mg/l of Pb, 0.85 mg/l of Fe and 0.35 mg/l of Cu in aerated aquaria for 96 hours. Tissue fractionation of the liver tissues was carried out and the three diagnostic enzymes (AST, ALT, and ALP) were estimated. Serum levels of the same diagnostic enzymes were also measured. The mean values of the serum enzyme activity for ALP in each experimental group were 19.5±1.62, 29.67±2.17 and 1.15±0.27 IU/L for Pb, Fe and Cu groups compared with 9.99±1.34 IU/L enzyme activity in the control. This result showed that Pb and Fe caused increased release of the enzyme into the blood circulation indicating increased tissue damage while Cu caused a reduction in the serum level as compared with the level in the control group. The mean values of enzyme activity obtained in the liver were 102.14±6.12, 140.17±2.06 and 168.23±3.52 IU/L for Pb, Fe and Cu groups, respectively compared to 91.20±9.42 IU/L enzyme activity for the control group. The serum and liver AST and ALT activities obtained in Pb, Fe, Cu and control groups are reported. It was generally noted that the presence of the heavy metal caused liver tissues damage and consequent increased level of the diagnostic enzymes in the serum.

Keywords: diagnostic enzymes, enzyme activity, heavy metals, tissues investigations

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Authors: Kira Rysakova, Vitaly Novikov

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An improved method of obtaining enzymatic collagen hydrolysate from the tissues of marine hydrobionts is proposed, which allows to obtain hydrolysate without pre-isolation of pure collagen. The method can be used to isolate enzymatic collagen hydrolysate from the waste of industrial processing of Red King crab and non-traditional objects - marine holothurias. Comparative analysis of collagen hydrolysates has shown the possibility of their use in a number of nutrient media, but this requires additional optimization of their composition and biological tests on wide sets of test strains of microorganisms.

Keywords: collagen hydrolysate, marine hydrobionts, red king crab, marine holothurias, enzymes, exclusive HPLC

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Authors: Sánchez Acuña, Juan Camilo, Granados Gómez, Mildred Magaly, Navarrete Rodríguez, Luisa Fernanda

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Nowadays, the main biofuels source production as bioethanol is food crops. This means a high competition between foods and energy production. For this reason, it is necessary to take into account the use of new raw materials friendly to the environment. The main objective of this paper is to evaluate the potential of the agro-industrial banana crop residues in the production of bioethanol. A factorial design of 2⁴ was used, the design has variables such as pH, time and concentration of hydrolysis, another variable is the time of fermentation that is of 7 or 15 days. In the hydrolysis phase, the pH is acidic (H₂SO₄) or basic (NaOH), the time is 30 or 15 minutes and the concentration is 0.1 or 0.5 M. It was observed that basic media, low concentrations, fermentation, and higher pretreatment times produced better performance in terms of biofuel obtained.

Keywords: bioethanol, biofuels, banana waste, hydrolysis

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Authors: Parastoo Motallebi, Vahid Niknam, Hassan Ebrahimzadeh, Majid Hashemi

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Wheat, the most strategically important worldwide crop, is widely grown in various countries. Based on international wheat production statistics (FAOSTAT database), the total production of wheat in 2012 was 13.8 in Iran. Fusarium culmorum is one of the principal causative agents of Fusarium crown rot (FCR), an overwhelming disease of wheat and barley which is in the early stages causing yield losses, stand reductions and rotting of root and lower stem tissues. In this study inoculation of two wheat seedlings of the susceptible cultivar Falat and the partially field-resistant cultivar Pishtaz were carried out in greenhouse conditions and root samples were taken for 6 days. The activity of peroxidase (POX) and polyphenoloxidase (PPO) enzymes were analyzed to identify possible relations between resistance and enzymatic activities. Although the POX and PPO activities in both geno types increased, this significant increase was more dominant in Pishtaz. The results showed an earlier elevation in the activity of these defense related enzymes in semi-resistant cv. Pishtaz after inoculation, suggested that the activities of POX and PPO in wheat geno types play an important role in the induction of resistance to this disease.

Keywords: Defense responses, Fusarium culmorum, Wheat

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Authors: Razieh Karimi Aghcheh, Christian Kubicek, Joseph Strauss, Gerhard Braus

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Filamentous fungi are of considerable economic and social significance for human health, nutrition and in white biotechnology. These organisms are dominant producers of a range of primary metabolites such as citric acid, microbial lipids (biodiesel) and higher unsaturated fatty acids (HUFAs). In particular, they produce also important but structurally complex secondary metabolites with enormous therapeutic applications in pharmaceutical industry, for example: cephalosporin, penicillin, taxol, zeranol and ergot alkaloids. Several fungal secondary metabolites, which are significantly relevant to human health do not only include antibiotics, but also e.g. lovastatin, a well-known antihypercholesterolemic agent produced by Aspergillus. terreus, or aflatoxin, a carcinogen produced by A. flavus. In addition to their roles for human health and agriculture, some fungi are industrially and commercially important: Species of the ascomycete genus Hypocrea spp. (teleomorph of Trichoderma) have been demonstrated as efficient producer of highly active cellulolytic enzymes. This trait makes them effective in disrupting and depolymerization of lignocellulosic materials and thus applicable tools in number of biotechnological areas as diverse as clothes-washing detergent, animal feed, and pulp and fuel productions. Fungal LaeA/LAE1 (Loss of aflR Expression A) homologs their gene products act at the interphase between secondary metabolisms, cellulase production and development. Lack of the corresponding genes results in significant physiological changes including loss of secondary metabolite and lignocellulose degrading enzymes production. At the molecular level, the encoded proteins are presumably methyltransferases or demethylases which act directly or indirectly at heterochromatin and interact with velvet domain proteins. Velvet proteins bind to DNA and affect expression of secondary metabolites (SMs) genes and cellulases. The dynamic interplay between LaeA/LAE1, velvet proteins and additional interaction partners is the key for an understanding of the coordination of metabolic and morphological functions of fungi and is required for a biotechnological control of the formation of desired bioactive products. Aspergilli and Trichoderma represent different biotechnologically significant species with significant differences in the LaeA/LAE1-Velvet protein machinery and their target proteins. We, therefore, performed a comparative study of the interaction partners of this machinery and the dynamics of the various protein-protein interactions using our robust proteomic and mass spectrometry techniques. This enhances our knowledge about the fungal coordination of secondary metabolism, cellulase production and development and thereby will certainly improve recombinant fungal strain construction for the production of industrial secondary metabolite or lignocellulose hydrolytic enzymes.

Keywords: cellulases, LaeA/1, proteomics, secondary metabolites

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Authors: Aleya Ferdausi, Meriel Jones, Anthony Halls

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The Amaryllidaceae genus Narcissus contains secondary metabolites, which are important sources of bioactive compounds such as pharmaceuticals indicating that their biological activity extends from the native plant to humans. Transcriptome analysis (RNA-seq) is an effective platform for the identification and functional characterization of candidate genes as well as to identify genes encoding uncharacterized enzymes. The biotechnological production of secondary metabolites in plant cell or organ cultures has become a tempting alternative to the extraction of whole plant material. The biochemical pathways for the production of secondary metabolites require primary metabolites to undergo a series of modifications catalyzed by enzymes such as cytochrome P450s, methyltransferases, glycosyltransferases, and acyltransferases. Differential gene expression analysis of Narcissus was obtained from two conditions, i.e. field and in vitro callus. Callus was obtained from modified MS (Murashige and Skoog) media supplemented with growth regulators and twin-scale explants from Narcissus cv. Carlton bulb. A total of 2153 differentially expressed transcripts were detected in Narcissus bulb and in vitro callus, and 78.95% of those were annotated. It showed the expression of genes involved in the biosynthesis of alkaloids were present in both conditions i.e. cytochrome P450s, O-methyltransferase (OMTs), NADP/NADPH dehydrogenases or reductases, SAM-synthetases or decarboxylases, 3-ketoacyl-CoA, acyl-CoA, cinnamoyl-CoA, cinnamate 4-hydroxylase, alcohol dehydrogenase, caffeic acid, N-methyltransferase, and NADPH-cytochrome P450s. However, cytochrome P450s and OMTs involved in the later stage of Amaryllidaceae alkaloids biosynthesis were mainly up-regulated in field samples. Whereas, the enzymes involved in initial biosynthetic pathways i.e. fructose biphosphate adolase, aminotransferases, dehydrogenases, hydroxyl methyl glutarate and glutamate synthase leading to the biosynthesis of precursors; tyrosine, phenylalanine and tryptophan for secondary metabolites were up-regulated in callus. The knowledge of probable genes involved in secondary metabolism and their regulation in different tissues will provide insight into the Narcissus plant biology related to alkaloid production.

Keywords: narcissus, callus, transcriptomics, secondary metabolites

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Authors: Carlos A. C. G. Neto, Natan C. G. e Silva , Thaís de O. Costa, Luciana R. B. Gonçalves, Maria V. P. Rocha

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β-galactosidases (E.C. 3.2.1.23) are enzymes that have attracted by catalyzing the hydrolysis of lactose and in producing galacto-oligosaccharides by favoring transgalactosylation reactions. These enzymes, when immobilized, can have some enzymatic characteristics substantially improved, and the coating of supports with multifunctional polymers is a promising alternative to enhance the stability of the biocatalysts, among which polyethylenimine (PEI) stands out. PEI has certain properties, such as being a flexible polymer that suits the structure of the enzyme, giving greater stability, especially for multimeric enzymes such as β-galactosidases. Besides that, protects them from environmental variations. The use of chitosan support coated with PEI could improve the catalytic efficiency of β-galactosidase from Kluyveromyces lactis in the transgalactosylation reaction for the production of prebiotics, such as lactulose since this strain is more effective in the hydrolysis reaction. In this context, the aim of the present work was first to develop biocatalysts of β-galactosidase from K. lactis immobilized on chitosan-coated with PEI, determining the immobilization parameters, its operational and thermal stability, and then to apply it in hydrolysis and transgalactolisation reactions to produce lactulose using whey as a substrate. The immobilization of β-galactosidase in chitosan previously functionalized with 0.8% (v/v) glutaraldehyde and then coated with 10% (w/v) PEI solution was evaluated using an enzymatic load of 10 mg protein per gram support. Subsequently, the hydrolysis and transgalactosylation reactions were conducted at 50 °C, 120 RPM for 20 minutes, using whey supplemented with fructose at a ratio of 1:2 lactose/fructose, totaling 200 g/L. Operational stability studies were performed in the same conditions for 10 cycles. Thermal stabilities of biocatalysts were conducted at 50 ºC in 50 mM phosphate buffer, pH 6.6 with 0.1 mM MnCl2. The biocatalyst whose support was coated was named CHI_GLU_PEI_GAL, and the one that was not coated was named CHI_GLU_GAL. The coating of the support with PEI considerably improved the parameters of immobilization. The immobilization yield increased from 56.53% to 97.45%, biocatalyst activity from 38.93 U/g to 95.26 U/g and the efficiency from 3.51% to 6.0% for uncoated and coated support, respectively. The biocatalyst CHI_GLU_PEI_GAL was better than CHI_GLU_GAL in the hydrolysis of lactose and production of lactulose, converting 97.05% of lactose at 5 min of reaction and producing 7.60 g/L lactulose in the same time interval. QUI_GLU_PEI_GAL biocatalyst was stable in the hydrolysis reactions of lactose during the 10 cycles evaluated, converting 73.45% lactose even after the tenth cycle, and in the lactulose production was stable until the fifth cycle evaluated, producing 10.95 g/L lactulose. However, the thermal stability of CHI_GLU_GAL biocatalyst was superior, with a half-life time 6 times higher, probably because the enzyme was immobilized by covalent bonding, which is stronger than adsorption (CHI_GLU_PEI_GAL). Therefore, the strategy of coating the supports with PEI has proven to be effective for the immobilization of β-galactosidase from K. lactis, considerably improving the immobilization parameters, as well as, the catalytic action of the enzyme. Besides that, this process can be economically viable due to the use of an industrial residue as a substrate.

Keywords: β-galactosidase, immobilization, kluyveromyces lactis, lactulose, polyethylenimine, transgalactosylation reaction, whey

Procedia PDF Downloads 97