Search results for: enzymes production

Authors: Zhanara B. Suleimenova, Zhazira K. Saduyeva

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Filamentous fungi are major producers of enzymes that have important applications in the food and beverage industries. The overall objective of this research is a strain improvement technology for efficient industrial enzymes production. The new way of filamentous fungi cultivation method has been developed. Such technology prolong producers’ cultivation period up to 60 days and create the opportunity to obtain enzymes repeatedly in every 2-3 days of fungal cultivation. This method is based on immobilizing enzymes producers with solid support in submerged conditions of growth. Immobilizing has a range of advantages: Decreasing the price of the final product, absence of foreign substances, controlled process of enzyme-genesis, ability of various enzymes simultaneous production, etc. Design of proposed technology gives the opportunity to increase the activity of immobilized cells culture filtrate comparing to free cells, growing in periodic culture conditions. Thus, proposed research focuses on new, more versatile, microorganisms capable of squeezing more end-products as well as proposed cultivation technology led to increased enzymatic productivity by several times.

Keywords: filamentous fungi, immobilization, industrial enzymes production, strain improvement

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Authors: Jwan J. Abdullah, Greetham Darren, Gregory A, Tucker, Chenyu Du

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Solid-state fermentation (SSF) is an alternative to liquid fermentations for the production of commercially important products such as antibiotics, single cell proteins, enzymes, organic acids, or biofuels from lignocellulosic material. This paper describes the optimisation of SSF on municipal solid waste (MSW) for the production of cellulase enzyme. Production of cellulase enzymes was optimised by Trichoderma reesei or Aspergillus niger for temperature, moisture content, inoculation, and period of incubation. Also, presence of minerals, and alternative carbon and nitrogen sources. Optimisation revealed that production of cellulolytic enzymes was optimal when using Trichoderma spp at 30°C with an incubation period of 168 hours with a 60% moisture content. Crude enzymes produced from MSW, by Trichoderma were evaluated for the saccharification of MSW and compared with activity of a commercially available enzyme, results demonstrated that MSW can be used as inexpensive lignocellulosic material for the production of cellulase enzymes using Trichoderma reesei.

Keywords: SSF, enzyme hydrolysis, municipal solid waste (MSW), optimizing conditions, enzyme hydrolysis

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Authors: Abeer A. Q. Ahmed, Tracey McKay

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The increasing demand for renewable fuels and chemicals is pressuring manufacturing industry toward finding more sustainable cost-effective resources. Lignocellulose, such as agro-food wastes, is a suitable equivalent to petroleum for fine chemicals and fuels production. The complex structure of lignocellulose, however, requires a variety of enzymes in order to degrade its components into their respective building blocks that can be used further for the production of various value added products. This study aimed to isolate bacterial strain with the ability to produce a variety of lignocellulosic enzymes. One bacterial isolate was identified by 16S rRNA gene sequencing and phylogenetic analysis as Bacillus safensis LCX found to have CMCase, xylanase, manganese peroxidase, lignin peroxidase, and laccase activities. The enzymes production was induced by growing Bacillus safensis LCX in solid state fermentation using wheat straw, wheat bran, and corn stover. The activities of enzymes were determined by specific colorimetric assays. This study presents Bacillus safensis LCX as a promising source for lignocellulosic enzymes. These findings can extend the knowledge on agro-food wastes valorization strategies toward a sustainable production of fuels and chemicals.

Keywords: Bacillus safensis LCX, high valued chemicals, lignocellulosic enzymes, solid state fermentation

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Authors: Ayuko Itsuki, Sachiyo Aburatani

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In recent years, pollutions of the environment by toxic substances become a serious problem. While there are many methods of environmental clean-up, the methods by microorganisms are considered to be reasonable and safety for environment. Compost is known that it catabolize the meladorous substancess in its production process, however the mechanism of its catabolizing system is not known yet. In the catabolization process, organic matters turn into inorganic by the released enzymes from lots of microorganisms which live in compost. In other words, the cooperative of activated enzymes in the compost decomposes malodorous substances. Thus, clarifying the interaction among enzymes is important for revealing the catabolizing system of meladorous substance in compost. In this study, we utilized statistical method to infer the interaction among enzymes. We developed a method which combined partial correlation with cross correlation to estimate the relevance between enzymes especially from time series data of few variables. Because of using cross correlation, we can estimate not only the associative structure but also the reaction pathway. We applied the developed method to the enzyme measured data and estimated an interaction among the enzymes in decomposition mechanism of toluene.

Keywords: enzyme activities, comparative analysis, compost, toluene

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Authors: Zhanara Suleimenova, Raushan Blieva, Aigerim Zhakipbekova, Inkar Tapenbayeva, Zhanar Narmuratova

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Green processes are based on innovative technologies that do not negatively affect the environment. Industrial enzymes originated from biological systems can effectively contribute to sustainable development through being isolated from microorganisms which are fermented using primarily renewable resources. Many widespread microorganisms secrete a significant amount of biocatalysts into the environment, which greatly facilitates the task of their isolation and purification. The ability to control the enzyme production through the regulation of their biosynthesis and the selection of nutrient media and cultivation conditions allows not only to increase the yield of enzymes but also to obtain enzymes with certain properties. In this regard, large potentialities are embedded in immobilized cells. Enzyme production technology in a secreted active form enabling industrial application on an economically feasible scale has been developed. This method is based on the immobilization of enzyme producers on a solid career. Immobilizing has a range of advantages: decreasing the price of the final product, absence of foreign substances, controlled process of enzyme-genesis, the ability of various enzymes' simultaneous production, etc. Design of proposed equipment gives the opportunity to increase the activity of immobilized cell culture filtrate comparing to free cells, growing in periodic culture conditions. Such technology allows giving a 10-times raise in culture productivity, to prolong the process of fungi cultivation and periods of active culture liquid generation. Also, it gives the way to improve the quality of filtrates (to make them more clear) and exclude time-consuming processes of recharging fermentative vials, that require manual removing of mycelium.

Keywords: industrial enzymes, immobilization, submerged fermentation, microscopic fungi

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Authors: Astorga-Trejo Rebeca, Fonseca-Peralta Héctor Manuel, Beltrán-Arredondo Laura Ivonne, Castro-Martínez Claudia

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The use of biomass as feedstock for the production of fuels and other chemicals of interest is an ever-growing accepted option in the way to the development of biorefinery complexes; in the Mexican state of Sinaloa, two million tons of residues from corn crops are produced every year, most of which can be converted to bioethanol and other products through biotechnological conversion using yeast and other microorganisms. Therefore, the objective of this work was to take advantage of corn stover and evaluate its potential as a substrate for the production of second-generation bioethanol (2G), enzymes, and xylitol. To produce bioethanol 2G, an acid-alkaline pretreatment was carried out prior to saccharification and fermentation. The microorganisms used for the production of enzymes, as well as for the production of xylitol, were isolated and characterized in our workgroup. Statistical analysis was performed using Design Expert version 11.0. The results showed that it is possible to obtain 2G bioethanol employing corn stover as a carbon source and Saccharomyces cerevisiae ItVer01 and Candida intermedia CBE002 with yields of 0.42 g and 0.31 g, respectively. It was also shown that C. intermedia has the ability to produce xylitol with a good yield (0.46 g/g). On the other hand, qualitative and quantitative studies showed that the native strains of Fusarium equiseti (0.4 IU/mL - xylanase), Bacillus velezensis (1.2 IU/mL – xylanase and 0.4 UI/mL - amylase) and Penicillium funiculosum (1.5 IU / mL - cellulases) have the capacity to produce xylanases, amylases or cellulases using corn stover as raw material. This study allowed us to demonstrate that it is possible to use corn stover as a carbon source, a low-cost raw material with high availability in our country, to obtain bioproducts of industrial interest, using processes that are more environmentally friendly and sustainable. It is necessary to continue the optimization of each bioprocess.

Keywords: biomass, corn stover, biorefinery, bioethanol 2G, enzymes, xylitol

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Authors: Wafaa M. Abd El-Rahim, Magda A. El-Meleigy, Eman Refaat

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This study dealing with optimization the conditions affecting the formation of extracellular lignin- degrading enzymes to achieve maximal decolorization activity of Direct Violet dye by one fungal strain. In this study Aspergillus fumigates fungal strain used for production extracellular ligninolytic enzymes for removing Direct Violet dye under different conditions: culture medium, incubation period, pH and temperatures. The results indicted that the removal efficiency of A. fumigatus was enhanced by addition glucose and peptone to the culture medium. The addition of peptone and glucose was found to increase the decolorization activity of the fungal isolate from 51.38% to 93.74% after 4 days of incubation. The highest production of extracellular lignin degrading enzymes also recorded in Direct Violet dye medium supplemented with peptone and glucose. It was also found the decolorization activity of A. fumigatus was decreased gradually by increasing the incubation period up to 4 days. Also it was found that the fungal strain can grow and produce extracellular ligninolytic enzymes which accompanied by efficient removal of Direct Violet dye in a wide pH range of 4-8. The results also found that the maximal biosynthesis of ligninolytic enzymes which accompanied with maximal removal of Direct Violet dye was obtained at a temperature of 28C. This indicates that the different conditions of culture medium, incubation period, pH and temperatures are effective on dye decolorization on the fungal biomass and played a role in Direct Violet dye removal along with enzymatic activity of A. fumigatus.

Keywords: A. fumigates, extracellular lignin- degrading enzymes, textile dye, dye removing

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Authors: V. Surena, B. Nazemisalman, F. Noghrehkar

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Introduction: Gingival overgrowth (GO) is a common side effect involving users of antiepileptic, immunosuppressive and calcium channel blocker drugs. Cyclosporine and phenytoin are amongst the most widely used drugs associated with GO. Gingival fibroblasts seem to have a significant role in the production of certain enzymes after administration of the drugs contributing to GO. Previous studies have shown a higher prevalence of GO in children and adolescents. The aim of this study was to compare normal human gingival fibroblasts with those exposed to Cyclosporine or phenytoin in measuring the production levels of certain enzymes that could have a possible role in GO. Methods: samples were obtained from the gingival biopsies of seven adult and seven children and were cultured into plates. With the growth of fibroblast cells, they were treated with or without either Cyclosporine or phenytoin. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to determine the expressed levels of R-EGF, cathepsin B,L, Lysyl oxidase, COL1, TGF β1, MMP-1,2, and TIMP1. Results: according to RT-PCR analyses, the expressed levels of R-EGF, cathepsin B, L, Lysyl oxidase, COL1, TGF β1, MMP-1, 2 and TIMP1 were affected by Cyclosporine and phenytoin. TGF-β1, TIMP, Cathepsin B and EGF showed comparable values in the adult and pediatric groups. Conclusions: Different expressed levels of enzymes after treatment of the gingival fibroblasts of adults and pediatrics with phenytoin or Cyclosporine could be the reason for the higher severity of GO in children. More studies need to be performed on the pathogenesis of GO at different age groups.

Keywords: cyclosporine, fibroblasts, phenytoin, gingivae

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Authors: Paula K. Novelli, Margarida M. Barros, Luciana F. Flueri

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Utilization of agricultural by-products, wheat ban and soybean bran, as substrate for solid state fermentation (SSF) was studied, aiming the achievement of different enzymes from Aspergillus sp. with distinct biological characteristics and its application and improvement on animal nutrition. Aspergillus niger and Aspergillus oryzea were studied as they showed very high yield of phytase and protease production, respectively. Phytase activity was measure using p-nitrophenilphosphate as substrate and a standard curve of p-nitrophenol, as the enzymatic activity unit was the quantity of enzyme necessary to release one μmol of p-nitrophenol. Protease activity was measure using azocasein as substrate. Activity for phytase and protease substantially increased when the different biochemical characteristics were considered in the study. Optimum pH and stability of the phytase produced by A. niger with wheat bran as substrate was between 4.0 - 5.0 and optimum temperature of activity was 37oC. Phytase fermented in soybean bran showed constant values at all pHs studied, for optimal and stability, but low production. Phytase with both substrates showed stable activity for temperatures higher than 80oC. Protease from A. niger showed very distinct behavior of optimum pH, acid for wheat bran and basic for soybean bran, respectively and optimal values of temperature and stability at 50oC. Phytase produced by A. oryzae in wheat bran had optimum pH and temperature of 9 and 37oC, respectively, but it was very unstable. On the other hand, proteases were stable at high temperatures, all pH’s studied and showed very high yield when fermented in wheat bran, however when it was fermented in soybean bran the production was very low. Subsequently the upscale production of phytase from A. niger and proteases from A. oryzae were applied as an enzyme additive in fish fed for digestibility studies. Phytases and proteases were produced with stable enzyme activity of 7,000 U.g-1 and 2,500 U.g-1, respectively. When those enzymes were applied in a plant protein based fish diet for digestibility studies, they increased protein, mineral, energy and lipids availability, showing that these new enzymes can improve animal production and performance. In conclusion, the substrate, as well as, the microorganism species can affect the biochemical character of the enzyme produced. Moreover, the production of these enzymes by SSF can be up to 90% cheaper than commercial ones produced with the same fungi species but submerged fermentation. Add to that these cheap enzymes can be easily applied as animal diet additives to improve production and performance.

Keywords: agricultural by-products, animal nutrition, enzymes production, solid state fermentation

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Authors: Shaohua Li, Aihua Zhang, Kelly Zatopek, Saba Parvez, Andrew F. Gardner, Ivan R. Corrêa Jr., Christopher J. Noren, Ming-Qun Xu

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Covalent immobilization of enzymes on solid supports is an alternative approach to biocatalysis with the added benefits of simple enzyme removal, improved stability, and adaptability to automation and high-throughput applications. Nevertheless, immobilized enzymes generally suffer from reduced activities compared to their soluble counterparts. One major factor leading to activity loss is the intrinsic hydrophobic property of the supporting material surface, which could result in the conformational change/confinement of enzymes. We report a strategy of utilizing flexible poly (ethylene oxide) (PEO) moieties as to improve the surface hydrophilicity of solid supports used for enzyme immobilization. DNA modifying enzymes were covalently conjugated to PEO-coated magnetic-beads. Kinetics studies proved that the activities of the covalently-immobilized DNA modifying enzymes were greatly enhanced by the PEO modification on the bead surface.

Keywords: immobilized enzymes, biocatalysis, poly(ethylene oxide), surface modification

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Authors: Nivedita Sharma

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The global demand for fossil fuels is very high, but their use is not sustainable since its reserves are declining. Additionally, fossil fuels are responsible for the accumulation of greenhouse gases. The emission of greenhouse gases from the transport sector can be reduced by substituting fossil fuels by biofuels. Thus, renewable fuels capable of sequestering carbon dioxide are in high demand. Second‐generation biofuels, which require lignocellulosic biomass as a substrate and ultimately producing ethanol, fall largely in this category. Bioethanol is a favorable and near carbon-neutral renewable biofuel leading to reduction in tailpipe pollutant emission and improving the ambient air quality. Lignocellulose consists of three main components: cellulose, hemicellulose and lignin which can be converted to ethanol with the help of microbial enzymes. Enzymatic hydrolysis of lignocellulosic biomass in 1st step is considered as the most efficient and least polluting methods for generating fermentable hexose and pentose sugars which subsequently are fermented to power alcohol by yeasts in 2nd step of the process. In the present technology, a complete bioconversion process i.e. potential hydrolytic enzymes i.e. cellulase and xylanase producing microorganisms have been isolated from different niches, screened for enzyme production, identified using phenotyping and genotyping, enzyme production, purification and application of enzymes for saccharification of different lignocellulosic biomass followed by fermentation of hydrolysate to ethanol with high yield is to be presented in detail.

Keywords: cellulase, xylanase, lignocellulose, bioethanol, microbial enzymes

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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: 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: Claudio Lamilla, Andrés Santos, Vicente Llanquinao, Jocelyn Hermosilla, Leticia Barrientos

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The industry in general is very interested in improving and optimizing industrial processes in order to reduce the costs involved in obtaining raw materials and production. Thus, an interesting and cost-effective alternative is the incorporation of bioactive metabolites in such processes, being an example of this enzymes which catalyze efficiently a large number of enzymatic reactions of industrial and biotechnological interest. In the search for new sources of these active metabolites, Antarctica is one of the least explored places on our planet where the most drastic cold conditions, salinity, UVA-UVB and liquid water available are present, features that have shaped all life in this very harsh environment, especially bacteria that live in different Antarctic ecosystems, which have had to develop different strategies to adapt to these conditions, producing unique biochemical strategies. In this work the production of cellulolytic enzymes of seven bacterial strains isolated from marine sediments at different sites in the Antarctic was evaluated. Isolation of the strains was performed using serial dilutions in the culture medium at M115°C. The identification of the strains was performed using universal primers (27F and 1492R). The enzyme activity assays were performed on R2A medium, carboxy methyl cellulose (CMC)was added as substrate. Degradation of the substrate was revealed by adding Lugol. The results show that four of the tested strains produce enzymes which degrade CMC substrate. The molecular identifications, showed that these bacteria belong to the genus Streptomyces and Pseudoalteromonas, being Streptomyces strain who showed the highest activity. Only some bacteria in marine sediments have the ability to produce these enzymes, perhaps due to their greater adaptability to degrade at temperatures bordering zero degrees Celsius, some algae that are abundant in this environment and have cellulose as the main structure. The discovery of new enzymes adapted to cold is of great industrial interest, especially for paper, textiles, detergents, biofuels, food and agriculture. These enzymes represent 8% of industrial demand worldwide and is expected to increase their demand in the coming years. Mainly in the paper and food industry are required in extraction processes starch, protein and juices, as well as the animal feed industry where treating vegetables and grains helps improve the nutritional value of the food, all this clearly puts Antarctic microorganisms and their enzymes specifically as a potential contribution to industry and the novel biotechnological applications.

Keywords: antarctic, bacteria, biotechnological, cellulolytic enzymes

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Authors: Dake Xiong, Ben Hankamer, Ian Ross

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The most urgent challenge of our time is to replace the depleting resources of fossil fuels by sustainable environmentally friendly alternatives. Hydrogen is a promising CO2-neutral fuel for a more sustainable future especially when produced photo-biologically. Hydrogen can be photosynthetically produced in unicellular green alga like Chlamydomonas reinhardtii, catalysed by the inducible highly active and bidirectional [FeFe]-hydrogenase enzymes (HydA). However, evolutionary and physiological constraints severely restrict the hydrogen yield of algae for industrial scale-up, mainly due to its competition among other metabolic pathways on photosynthetic electrons. Among them, a major challenge to be resolved is the inferior competitiveness of hydrogen production (catalysed by HydA) with NADPH production (catalysed by ferredoxin-NADP+-reductase (FNR)), which is essential for cell growth and takes up ~95% of photosynthetic electrons. In this work, the in vivo hydrogen production efficiency of mutants with ferredoxin-hydrogenase (Fd*-HydA1*) fusion protein construct, where the electron donor ferredoxin (Fd*) is fused to HydA1* and expressed in the model organism C. reinhardtii was investigated. Once Fd*-HydA1* fusion gene is expressed in algal cells, the fusion enzyme is able to draw the redistributed photosynthetic electrons and use them for efficient hydrogen production. From preliminary data, mutants with Fd*-HydA1* transgene showed a ~2-fold increase in the photosynthetic hydrogen production rate compared with its parental strain, which only possesses the native HydA in vivo. Therefore, a solid method of having more efficient hydrogen production in microalgae can be achieved through the expression of the synthetic enzymes.

Keywords: Chlamydomonas reinhardtii, ferredoxin, fusion protein, hydrogen production, hydrogenase

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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: 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: Elsayed Ahmed Elsayed, Azza Noor El-Deen, Mohamed A. Farid, Mohamed A. Wadaan

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Recently, a great attention has been paid to the use of dietary carbohydrates as prebiotic functional foods. Among the new commercially available products, fructooligosaccharides (FOS), which are microbial produced from sucrose, have attracted special interest due to their valuable properties and, thus, have a great economic potential for the sugar industrial branch. They are non-cariogenic sweeteners of low caloric value, as they are not hydrolyzed by the gastro-intestinal enzymes, promoting selectively the growth of the bifidobacteria in the colon, helping to eliminate the harmful microbial species to human and animal health and preventing colon cancer. FOS has been also found to reduce cholesterol, phospholipids and triglyceride levels in blood. FOS has been mainly produced by microbial fructosyltransferase (FTase) enzymes. The present work outlines bioprocess optimization for different cultivation parameters affecting the production of FTase by Penicillium aurantiogriseum AUMC 5605. The optimization involves both traditional as well as fractional factorial design approaches. Additionally, the production process will be compared under batch and fed-batch conditions. Finally, the optimized process conditions will be applied to 5-L stirred tank bioreactor cultivations.

Keywords: prebiotics, fructooligosaccharides, optimization, cultivation

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Authors: Parmjit S. Panesar, Rupinder Kaur, Ram S. Singh

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Enzymes are the biocatalysts which catalyze the biochemical processes and thus have a wide variety of applications in the industrial sector. β-Galactosidase (E.C. 3.2.1.23) also known as lactase, is one of the prime enzymes, which has significant potential in the dairy and food processing industries. It has the capability to catalyze both the hydrolytic reaction for the production of lactose hydrolyzed milk and transgalactosylation reaction for the synthesis of prebiotics such as lactulose and galactooligosaccharides. These prebiotics have various nutritional and technological benefits. Although, the enzyme is naturally present in almonds, peaches, apricots and other variety of fruits and animals, the extraction of enzyme from these sources increases the cost of enzyme. Therefore, focus has been shifted towards the production of low cost enzyme from the microorganisms such as bacteria, yeast and fungi. As compared to yeast and bacteria, fungal β-galactosidase is generally preferred as being extracellular and thermostable in nature. Keeping the above in view, the present study was carried out for the isolation of the β-galactosidase producing fungal strain from the food as well as the agricultural wastes. A total of more than 100 fungal cultures were examined for their potential in enzyme production. All the fungal strains were screened using X-gal and IPTG as inducers in the modified Czapek Dox Agar medium. Among the various isolated fungal strains, the strain exhibiting the highest enzyme activity was chosen for further phenotypic and genotypic characterization. The strain was identified as Rhizomucor pusillus on the basis of 5.8s RNA gene sequencing data.

Keywords: beta-galactosidase, enzyme, fungal, isolation

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Authors: Golnoosh Torabian, P. Valtchev, F. Dehghani

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The need for efficient nutraceutical products has been dramatically changing the approach of the industrial processes. The development of novel mild processes is highly demanded for the production of such products; especially when both quality and safety need to be guaranteed during their long shelf life. Within this research, for the first time, we investigated the effect of supercritical carbon dioxide treatment for the inactivation of microbes and enzymes in a berry juice possessing therapeutic effect. We demonstrated that a complete inactivation of microbes can be achieved at optimized conditions of treatment. However, the bottle neck of the process was represented by the unpromising inactivation of the degradative enzyme by supercritical carbon dioxide treatment. However, complete enzyme inactivation was achieved by applying two strategies: the first was optimizing juicing method by adding a mechanical step and the second strategy was addition of natural inhibitors to the juice. Overall these results demonstrate that our hybrid process has a significant effect on the inactivation of microorganism and enzymes in the fresh juice. The developed process opens the possibility for the evolution of new products with optimal nutritional and sensorial characteristics, as well as offering a competitive cost and an environmentally friendly alternative for pasteurization and extension of shelf life in a wide range of natural therapeutic products.

Keywords: hybrid method, berry juice, pasteurization, enzymes inactivation

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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

Abstract:

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: Sidra Pervez, Afsheen Aman, Shah Ali Ul Qader

Abstract:

The importance of attaining an optimum performance of an enzyme is often a question of devising an effective method for its immobilization. Cross-linked enzyme aggregate (CLEAs) is a new approach for immobilization of enzymes using carrier free strategy. This method is exquisitely simple (involving precipitation of the enzyme from aqueous buffer followed by cross-linking of the resulting physical aggregates of enzyme molecules) and amenable to rapid optimization. Among many industrial enzymes, amyloglucosidase is an important amylolytic enzyme that hydrolyzes alpha (1→4) and alpha (1→6) glycosidic bonds in starch molecule and produce glucose as a sole end product. Glucose liberated by amyloglucosidase can be used for the production of ethanol and glucose syrups. Besides this amyloglucosidase can be widely used in various food and pharmaceuticals industries. For production of amyloglucosidase on commercial scale, filamentous fungi of genera Aspergillus are mostly used because they secrete large amount of enzymes extracellularly. The current investigation was based on isolation and identification of filamentous fungi from genus Aspergillus for the production of amyloglucosidase in submerged fermentation and optimization of cultivation parameters for starch saccharification. Natural isolates were identified as Aspergillus niger KIBGE-IB36, Aspergillus fumigatus KIBGE-IB33, Aspergillus flavus KIBGE-IB34 and Aspergillus terreus KIBGE-IB35 on taxonomical basis and 18S rDNA analysis and their sequence were submitted to GenBank. Among them, Aspergillus fumigatus KIBGE-IB33 was selected on the basis of maximum enzyme production. After optimization of fermentation conditions enzyme was immobilized on CLEA. Different parameters were optimized for maximum immobilization of amyloglucosidase. Data of enzyme stability (thermal and Storage) and reusability suggested the applicability of immobilized amyloglucosidase for continuous saccharification of starch in industrial processes.

Keywords: aspergillus, immobilization, industrial processes, starch saccharification

Procedia PDF Downloads 462

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

Abstract:

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

Procedia PDF Downloads 394

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

Abstract:

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: Ehsan Khorshidian, Afshin Farahbakhsh, Sina Aghili

Abstract:

Enzymes are promising biocatalysts for many organic reactions. They have excellent features like high activity, specificity and selectivity, and can catalyze under mild and environment friendly conditions. Epoxy-activated supports are almost-ideal ones to perform very easy immobilization of proteins and enzymes at both laboratory and industrial scale. The activated epoxy supports (chitosan/alginate, Eupergit C) may be very suitable to achieve the multipoint covalent attachment of proteins and enzymes, therefore, to stabilize their three-dimensional structure. The enzyme is firstly covalently immobilized under conditions pH 7.0 and 10.0. The remaining groups of the support are blocked to stop additional interaction between the enzyme and support by mercaptoethanol or Triton X-100. The results show support allowed obtaining biocatalysts with high immobilized protein amount and hydrolytic activity. The immobilization of lipases on epoxy support may be considered as attractive tool for obtaining highly active biocatalysts to be used in both aqueous and anhydrous aqueous media.

Keywords: immobilization of enzymes, epoxy supports, enzyme multipoint covalent attachment, microbial lipases

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Authors: Reshmi Vijayan

Abstract:

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

Abstract:

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

Procedia PDF Downloads 260