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Mikotoksinų kaupimosi koncentruotuose pašaruose tyrimų analizė / Analysis of mycotoxins accumuliation in concentrated fodderLebedevaitė, Erika 08 April 2005 (has links)
Aim of study. The purpose was to investigate and analyse mycotoxicological contamination of concentrated fodder during 2 years and to evaluate the use of chemical conserving CALPRONA NC.
Main tasks:
1) To investigate the amounts of aflatoxin B1, zearelenone, T2 and deoxivalenol (DON) mycotoxins in fodder using imunoferment analysis and chromatography (TLC) method’s.
2) To determine minimum and maximum levels of mycotoxins in fodder and compare with the formative’s.
3) Compare levels of mycotoxins in fodder during period of two years.
4) Investigate the influence of chemical conserving CALPRONA NC mycostatical effect on the grain after the harvest.
Material and methods:
Conservated (CALPRONA NC) and non conservated samples were taken from experimental farm.
Quantitative contamination of fungi (cfu – colony forming units) was determined using dilution method and Czapec Dox agar (Курасова, 1971).
Level of mycotoxins was determined using IFA (imunoferment analyses) method with commercial set VERATOX®DON, T- 2 toxin, Zearalenone. Also the TLC method was applied using silicon gel plates (methology of Romer Lab.)
Diffusion of mycotoxins in fodder during year 2003- 2004 was determined using processing of facts with “R” statistical pack.
Results and discussion:
1) Probe fodder is contaminated with fungi and their toxins: DON, zearalenone, T- 2 and aflatoxin B1.
2) Highest levels of mycotoxins in concentrated fodder (mg/kg or ppm) are DON (0.095 ppm), comparing to zearalenone (ZON) and... [to full text]
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The effect of vesicular-arbuscular mycorrhiza on the growth of two indigenous grass species Themeda triandra and Trachypogon spicatus grown on coalmine spoil topsoil.Lee, Alan. 23 December 2013 (has links)
The main project was an assessment of the effect that colonization by five different
Vesicular-arbuscular mycorrhiza (VAM) cultures have on the growth of the indigenous-grasses
Themeda triandra and Trachypogon spicatus, when grown on coalmine topsoil.
With unamended topsoil, VAM showed the ability to significantly increase the growth of
the grasses compared to non-VAM control plants. The amount of effect varied with the VAM
inoculum culture type, with a VAM culture originally from the Cape Flats being the most
effective. In a second trial, soil fertilized with nitrogen, potassium and low concentrations of
phosphate (P) was used. Again VAM displayed the ability to improve grass plant growth. The
increase in P caused the Large spore inoculum to become the most effective. This indicated that
different VAM cultures are inhibited to different degrees by an increase in phosphate fertilization.
The low level of VAM infection, in both trials, seemed to preclude most of the VAM associated nutrient uptake control.
Varying reports have been published on the effect of fertilization on VAM infection and
colonization. In an attempt to further elucidate the role of fertilizer in VAM inhibition,
rhizosphere soil from a long term fertility trial near Witbank, S.A. was sampled. Amcoal
environmental services fertilized forty-two plots with varying concentrations of nitrogen,
potassium, phosphate and lime to assess the growth of a variety of grasses. The trial had been
maintained for ten years before sampling was completed for this project. Samples from each plot
were taken from the rhizosphere soil of the most prominent grass (Digitaria eriantha). VAM
spores were extracted from all the samples and five different types of spores were identified and
counted for each sample. By comparing spore counts from each plot, the effect that the fertilizer
regime had on the VAM on that plot could be assessed.
Variation in the concentrations of nitrogen (N) and potassium had no significant effect on
VAM colonization. Very low concentrations of N could not be assessed as all plots had been
initially top dressed with nitrogen fertilizer. Phosphate (P) fertilizer concentration had a marked
effect on spore concentrations. There was a significant increase in spore concentration as P levels
were increased from zero P fertilization to 80kgs P/ha. Further increase in P to ≥ 60kgs P/ha
resulted in a significant decrease in spore concentrations. From this it would appear that a low
level of soil P is needed to give maximum VAM colonization and further increase in soil P causes
VAM inhibition. Lime ameliorated the VAM inhibition caused by high concentrations of P.
Increase in P caused spore concentrations of low abundance propagules (LAP) too decreased
more rapidly than high abundance propagules (HAP). In high P soils VAM with LAP would
eventually be eliminated from the system resulting in a decrease in VAM diversity.
A project was attempted to use the recently developed Randomly Amplified Polymorphic
DNA in conjunction with the Polymerase Chain Reaction (RAPD PCR) techniques to identify different VAM families. The technique causes the amplification of segments of DNA which can
be visualized by gel electrophoresis and staining. Band patterns formed can be related to the VAM
of origin and hence used in identification of that VAM. An attempt was made to amplify DNA
from a single spore in this manner which would, in conjunction with morphological observations,
make identification of VAM easier and more accurate. Problems with either releasing the DNA
from the spores, or substances in the spore inhibiting the PCR reaction made obtaining band
patterns difficult. After many PCR attempts, varying extraction methods and PCR conditions, no repeatable results could be obtained and work on this project was discontinued. / Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 1997.
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Oleoegelio su čiobrelių eteriniu aliejumi sudėties moduliavimas ir kokybės vertinimas / The Composition Modeling and Quality Assessment of Oleogel With Thyme Essential OilStukaitė, Aurelija 30 June 2014 (has links)
Pėdų grybelis (lot. Tinea pedis) – tai dermatofitų Trichophyton rubrum, Trichophyton mentagrophytes ir Epidermophyton floccosum sukeltas lėtinis pėdų, kojų tarpupirščių pažeidimas, pasireiškiantis odos įtrūkimais, pleiskanojimu, nemaloniu kvapu ir niežuliu. Vaistinio čiobrelio (lot. Thymus vulgaris L.) eterinis aliejus, savyje turintis timolio (36 – 55 proc.), gali apsaugoti pėdas nuo grybelio atsiradimo. Čiobrelių eterinį aliejų patogu įvesti į aliejinę terpę, tačiau aliejinius tirpalus naudoti nėra patogu. Todėl šio darbo tikslas pagaminti lipofilinį gelį, skirtą pėdų grybelio prevencijai.
Šis tyrimas apima: 1) timolio kiekio nustatymą čiobrelių eteriniame aliejuje; 2) optimalaus aliejaus ir gelifikuojančios medžiagos santykio parinkimą; 3) pagaminto lipofilinio gelio su čiobrelių eteriniu aliejumi klampos, pH reikšmės ir išvaizdos įvertinimą; 4) oleogelio veiksmingumo nustatymą, atliekant mikrobiologinį tyrimą.
Rezultatai: ESC būdu nustatyta, kad čiobrelių eteriniame aliejuje timolio yra 41,06±0,03 proc. Oleogelio pagrindu nuspręsta naudoti alyvuogių aliejų, nes skystojo parafino ir alyvuogių aliejaus mišinys nesudarė stabilios gelinės struktūros. Pagaminti oleogeliai visą stebėjimo laiką (6 mėn.) išlaikė savo išvaizdą – vientisą struktūrą, skaidrumą, šviesiai geltoną spalvą, čiobrelių kvapą ir pusiau kietą konsistenciją. Atlikus pH reikšmės stebėjimus nustatyta, kad oleogelių pH reikšmė reikšmingai nekito ir išliko atitinkanti žmogaus fiziologinę odos pH reikšmę... [toliau žr. visą tekstą] / Foot fungus (lat. Tinea pedis) is a skin disease caused by dermatophytes Trichophyton rubrum, Trichophyton mentagrophytes and Epidermophyton floccosum. The symptoms are cracks, itch, dandruff and unpleasant odor of the feet skin. Thyme (lat. Thymus vulgaris L.) essential oil containing of 36 – 55 percent of thymol can protect feet from fungus. Thyme essential oil is convenient to introduce in oil medium, but oil solutions are uncomfortable to use. Therefore, the aim of this work is to produce a lipophilic gel for prevention of foot fungus.
This study includes the following: 1) determining the amount of thymol in the thyme essential oil; 2) determination of the optimal ratio of oil and gelling agent material; 3) pH value and appearance evaluation of the produced lipophilic gel with thyme essential oil viscosity; 4) the effectiveness of the lipophilic gel in microbiological research.
The results: using HPLC method it has been found out that thyme essential oil contains 41.06±0.03 percent thymol. It was decided to use olive oil on oleogel basis because liquid paraffin and olive oil mixture didn’t make stable gel structure. Produced oleogels kept the same appearance features for all observation time (6 months) such as homogenous structure, clearness, light yellow color, the smell of thyme and semi – solid consistency. During the measurement of pH values of oleogels, it has been found out that pH values matched human physiological pH value. During the measurement of viscosity of... [to full text]
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Production And Biochemical Characterization Of Polyphenol Oxidase From Thermomyces LanuginosusAstarci, Erhan 01 January 2003 (has links) (PDF)
Polyphenol oxidases are enzymes that catalyze the oxidation of certain phenolic substrates to quinones in the presence of molecular oxygen. Polyphenol oxidases are widely used in several applications. In food industry, they are used for enhancement of flavor in coffee, tea and cocoa production, and determination of food quality. In medicine, they have several uses in treatments of Parkinson&rsquo / s disease, phenlyketonurea and leukemia. In wastewater treatment, they are used for the removal of phenolic pollutants from wastewaters. In pharmaceutical industry, differentiation of morphine from codeine is possible by means of polyphenol oxidase immobilized electrodes. In this study, a thermophilic fungus, Thermomyces lanuginosus was evaluated in terms of poyphenol oxidase production. The effect of different nutrient sources, inducers and fermentation parameters on enzyme production were investigated and maximum PPO activity of 97 U/ml was observed in bioreactor experiments at 50° / C, 400 rpm and pH 8.0 in a fermentation medium containing 1.4% yeast extract, 0.3% MgSO4, 1% KH2PO4, 0.003% CuSO4, 0.032% gallic acid. Type of polyphenol oxidase produced by Thermomyces lanuginosus was determined as laccase. For biochemical characterization studies, the enzyme was enriched by electrophoresis. Temperature and pH optima for the enzyme were determined as 60° / C and 8.0, respectively. Enzyme retained 67% activity after 1 h incubation at 80° / C and retained 87% of its activity after 1 hour of incubation at pH 9.0 at room temperature. The enzyme obeys Michealis-Menten kinetics with Km and Vmax values being 5 mg /ml catechol and 38 U/ml, respectively. Molecular weight of the enzyme was determined as 29 kDa and isoelectric point of enzyme was found to be approximately 6.0.
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Studies on charcoal rot of mungbeanFuhlbohm, Michael John Unknown Date (has links)
The fungus Macrophomina phaseolina is the causal agent of several diseases of mungbean (Vigna radiata). One of these diseases, known as charcoal rot, causes spoilage of germinating seed lots, and occurs when seed contaminated with M. phaseolina is used for sprouting. The detection of the pathogen during seed testing prior to export leads to downgrading of the seed and a resultant financial penalty to the grain grower. The aims of this research were: to determine the location of M. phaseolina in diseased and symptomless tissue of mungbean plants; to determine the mode, site and timing of seed colonisation of mungbean; to determine the impact of biotic and abiotic factors on seed colonisation; to conduct an assessment of genotypic variation of M. phaseolina within single plants; to determine modes of transport of inoculum that contribute to foliage infection and seed colonisation of mungbean; and to assess both pre- and post-harvest management strategies in order to reduce or prevent seed colonisation, or to minimise the process of seed transmission. The location of M. phaseolina in mungbean plants was determined through serial sectioning of, and subsequent isolation from, naturally infected host tissue. A large proportion of the mungbean tissue infected by M. phaseolina was found to be symptomless. Moreover, there were large areas of ostensibly pathogen-free tissue separating infection foci, thus indicating a strong likelihood of independent aerial infections. A selection of 14 isolates obtained from serial sectioning were assessed for genotypic variation with six primers using RAPD analysis. Of the 36 bands that were scored, 78% were polymorphic and as a result, 12 distinct genotypes were detected. Polymorphisms were also detected amongst isolates obtained from the same discrete infection area on single plants, which strongly suggests the occurrence of multiple aerial infections. Various methods of controlled inoculation including soil infestation, pod and foliar inoculations, and artificial seed infestation, were used to determine how mungbean seeds are colonised by M. phaseolina. Additionally, most of these inoculation methods were coupled with a series of abiotic treatments (temperature regimes, watering regimes, application of herbicides) that were designed to initiate stress conditions within infected plants, and possibly trigger growth of M. phaseolina from the infection courts and colonise seed. Seed colonisation was established in vitro and in vivo when immature pods were directly inoculated with microsclerotia of M. phaseolina. At least two days exposure at 100% relative humidity (RH) was necessary to establish seed infection in detached mungbean pods that were inoculated with microsclerotia of M. phaseolina. Extensive seed infection was still obtained when one or more days of 100% RH was interrupted by up to three days at low humidity. All except one of the other methods of controlled inoculation failed to produce colonised seed even when combinations of stresses were applied. Only when the bipyridylium herbicide Spray Seed 250 was applied to plants following the inoculation of mungbean stems within 13 cm of the pods, was seed colonised by M. phaseolina. This result raises the possibility that delayed harvesting of desiccated mungbean crops may promote further colonisation of mungbean tissue, including seed. Very strong evidence for the colonisation of mungbean seeds after deposition of soil-splashed inoculum of M. phaseolina onto pods was obtained through field and laboratory-based studies. Soil-splashed inoculum (most likely microsclerotia) was also found to be the source of inoculum responsible for the development of Macrophomina leaf blight of mungbean at several regional sites. To further investigate this finding, areas of several mungbean crops growing in naturally infested soil were covered in hessian cloth to prevent soil-splash and assessments of the levels of seed colonisation between covered and uncovered areas were made. Although colonisation of seed in the covered areas was significantly lower than in the uncovered areas, covering the soil did not eliminate colonisation of seed. This result suggests that inoculum dispersal, leading to pod infection and subsequent seed colonisation, occurs not only in splashed-soil but also by other means. Soil transported to the extra-floral nectaries of mungbeans by ants was found to contain infective inoculum of M. phaseolina. Furthermore, air-borne debris and dust collected in a trap contained viable microsclerotia of the pathogen. Isolates of M. phaseolina collected from both sources were pathogenic on mungbean seedlings, and suspensions of ant-transported soil and air-borne dust/debris infected mungbean pods and seed. This is the first report of both modes of inoculum dispersal. All three modes undoubtedly contribute to the total level of colonised seed, but their relative importance remains to be determined. Several options for the management of charcoal rot in mungbean seeds were investigated. Application of the fungicide carbendazim to mungbean plants after flowering significantly decreased, but did not prevent, colonisation of mungbean seed by M. phaseolina. Consequently, this method of management holds little promise for mungbean growers. A large number of weeds common in Australian mungbean fields were newly reported as hosts of M. phaseolina. Isolates obtained from the infected, but symptomless weeds were pathogenic on mungbean seedlings, thus indicating a lack of host-specificity toward mungbean. It is strongly suspected that the use of herbicides to control weeds in reduced and zero-tillage farming systems is increasing the risk of infection in subsequent mungbean crops through the build-up of inoculum in soil. This increased risk of infection may be further exacerbated by retaining stubble infested with M. phaseolina on the soil surface, thereby increasing the amount of inoculum that could be splashed onto plant organs. Surface sterilisation, using sodium hypochlorite, significantly reduced colonisation levels in heavily colonised mungbean seed lines, but the process was not enhanced when a partial-vacuum was introduced to the process. In some seed lots, up to 32% of the seed was colonised only on the seed coat (defined here as contamination), whereas the remainder was internally infected. Surface sterilisation also reduced the overall colonisation of 132 commercial lines by approximately one-third. Storage of seed for one month at either 4°C or 15°C significantly reduced colonisation levels in seed, whereas freezing treatments did not. Eradication of M. phaseolina from mungbean seed was possible through thermotherapy. However, the conditions required for eradication also contributed to large increases in abnormal germination levels and large losses in overall germination - an unacceptable trade-off for sprouters. A combination of thermotherapy and surface sterilisation of colonised mungbean seed may provide a more efficient process of seed treatment.
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Studies on charcoal rot of mungbeanFuhlbohm, Michael John Unknown Date (has links)
The fungus Macrophomina phaseolina is the causal agent of several diseases of mungbean (Vigna radiata). One of these diseases, known as charcoal rot, causes spoilage of germinating seed lots, and occurs when seed contaminated with M. phaseolina is used for sprouting. The detection of the pathogen during seed testing prior to export leads to downgrading of the seed and a resultant financial penalty to the grain grower. The aims of this research were: to determine the location of M. phaseolina in diseased and symptomless tissue of mungbean plants; to determine the mode, site and timing of seed colonisation of mungbean; to determine the impact of biotic and abiotic factors on seed colonisation; to conduct an assessment of genotypic variation of M. phaseolina within single plants; to determine modes of transport of inoculum that contribute to foliage infection and seed colonisation of mungbean; and to assess both pre- and post-harvest management strategies in order to reduce or prevent seed colonisation, or to minimise the process of seed transmission. The location of M. phaseolina in mungbean plants was determined through serial sectioning of, and subsequent isolation from, naturally infected host tissue. A large proportion of the mungbean tissue infected by M. phaseolina was found to be symptomless. Moreover, there were large areas of ostensibly pathogen-free tissue separating infection foci, thus indicating a strong likelihood of independent aerial infections. A selection of 14 isolates obtained from serial sectioning were assessed for genotypic variation with six primers using RAPD analysis. Of the 36 bands that were scored, 78% were polymorphic and as a result, 12 distinct genotypes were detected. Polymorphisms were also detected amongst isolates obtained from the same discrete infection area on single plants, which strongly suggests the occurrence of multiple aerial infections. Various methods of controlled inoculation including soil infestation, pod and foliar inoculations, and artificial seed infestation, were used to determine how mungbean seeds are colonised by M. phaseolina. Additionally, most of these inoculation methods were coupled with a series of abiotic treatments (temperature regimes, watering regimes, application of herbicides) that were designed to initiate stress conditions within infected plants, and possibly trigger growth of M. phaseolina from the infection courts and colonise seed. Seed colonisation was established in vitro and in vivo when immature pods were directly inoculated with microsclerotia of M. phaseolina. At least two days exposure at 100% relative humidity (RH) was necessary to establish seed infection in detached mungbean pods that were inoculated with microsclerotia of M. phaseolina. Extensive seed infection was still obtained when one or more days of 100% RH was interrupted by up to three days at low humidity. All except one of the other methods of controlled inoculation failed to produce colonised seed even when combinations of stresses were applied. Only when the bipyridylium herbicide Spray Seed 250 was applied to plants following the inoculation of mungbean stems within 13 cm of the pods, was seed colonised by M. phaseolina. This result raises the possibility that delayed harvesting of desiccated mungbean crops may promote further colonisation of mungbean tissue, including seed. Very strong evidence for the colonisation of mungbean seeds after deposition of soil-splashed inoculum of M. phaseolina onto pods was obtained through field and laboratory-based studies. Soil-splashed inoculum (most likely microsclerotia) was also found to be the source of inoculum responsible for the development of Macrophomina leaf blight of mungbean at several regional sites. To further investigate this finding, areas of several mungbean crops growing in naturally infested soil were covered in hessian cloth to prevent soil-splash and assessments of the levels of seed colonisation between covered and uncovered areas were made. Although colonisation of seed in the covered areas was significantly lower than in the uncovered areas, covering the soil did not eliminate colonisation of seed. This result suggests that inoculum dispersal, leading to pod infection and subsequent seed colonisation, occurs not only in splashed-soil but also by other means. Soil transported to the extra-floral nectaries of mungbeans by ants was found to contain infective inoculum of M. phaseolina. Furthermore, air-borne debris and dust collected in a trap contained viable microsclerotia of the pathogen. Isolates of M. phaseolina collected from both sources were pathogenic on mungbean seedlings, and suspensions of ant-transported soil and air-borne dust/debris infected mungbean pods and seed. This is the first report of both modes of inoculum dispersal. All three modes undoubtedly contribute to the total level of colonised seed, but their relative importance remains to be determined. Several options for the management of charcoal rot in mungbean seeds were investigated. Application of the fungicide carbendazim to mungbean plants after flowering significantly decreased, but did not prevent, colonisation of mungbean seed by M. phaseolina. Consequently, this method of management holds little promise for mungbean growers. A large number of weeds common in Australian mungbean fields were newly reported as hosts of M. phaseolina. Isolates obtained from the infected, but symptomless weeds were pathogenic on mungbean seedlings, thus indicating a lack of host-specificity toward mungbean. It is strongly suspected that the use of herbicides to control weeds in reduced and zero-tillage farming systems is increasing the risk of infection in subsequent mungbean crops through the build-up of inoculum in soil. This increased risk of infection may be further exacerbated by retaining stubble infested with M. phaseolina on the soil surface, thereby increasing the amount of inoculum that could be splashed onto plant organs. Surface sterilisation, using sodium hypochlorite, significantly reduced colonisation levels in heavily colonised mungbean seed lines, but the process was not enhanced when a partial-vacuum was introduced to the process. In some seed lots, up to 32% of the seed was colonised only on the seed coat (defined here as contamination), whereas the remainder was internally infected. Surface sterilisation also reduced the overall colonisation of 132 commercial lines by approximately one-third. Storage of seed for one month at either 4°C or 15°C significantly reduced colonisation levels in seed, whereas freezing treatments did not. Eradication of M. phaseolina from mungbean seed was possible through thermotherapy. However, the conditions required for eradication also contributed to large increases in abnormal germination levels and large losses in overall germination - an unacceptable trade-off for sprouters. A combination of thermotherapy and surface sterilisation of colonised mungbean seed may provide a more efficient process of seed treatment.
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The growth response of Eucalyptus grandis x E. camaldulensis to salt stress, ectomycorrhizae and endomycorrhizae double colonisation /Hengari, Simeon Ngaitungue. January 2007 (has links)
Thesis (MScBosb)--University of Stellenbosch, 2007. / Bibliography. Also available via the Internet.
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Changes in growth and survival by three co-occurring grass species in response to mycorrhizae, fire, and droughtWilkinson, Melinda M. January 2003 (has links)
Thesis (Ph. D.)--University of Hawaii at Manoa, 2003. / Includes bibliographical references.
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Predators associated with hemlock woolly adelgid (Hemiptera: Adelgidae) infested western hemlock in the Pacific Northwest /Kohler, Glenn R. January 1900 (has links)
Thesis (M.S.)--Oregon State University, 2007. / Printout. Includes bibliographical references. Also available on the World Wide Web.
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Produção de poligalacturonase termoestável pelo fungo Rhizomucor pusillus A 13.36 em fermentação em estado sólido, purificação e caracterização da enzimaFreitas, Paula Mendes de [UNESP] 27 November 2009 (has links) (PDF)
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freitas_pm_dr_rcla.pdf: 670514 bytes, checksum: 22df87c8cdf056a03825df30dc778c89 (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / O fungo Rhizomucor pusillus A 13.36, quando cultivado por fermentação em estado sólido utilizando farelo de algodão como fonte de carbono por 72 horas, produziu altos níveis de poligalacturonase (18,6U/g). Essa enzima apresentou atividade máxima em pH 5,5 e em 55oC. A poligalacturonase bruta apresentou estabilidade na faixa de pH entre 4,5 e 6,5, com uma atividade residual de 91,3% no pH 6,5, e também até 70oC por 1h, com atividade residual de 80% nas temperaturas de 55 a 60oC. Esta enzima foi purificada por concentração e uma combinação de procedimentos de gel filtração e cromatografia de troca-iônica. A massa molecular da enzima foi 53,7kDa. A focalização isoelétrica mostrou uma única banda cujo ponto isoelétrico (pI) foi 3,8. A atividade máxima da enzima purificada também ocorreu em pH 5,5 e a 55oC. A enzima purificada foi estável na faixa de pH 4,5 a 6,5, com atividade residual na faixa de 80 a 100%, e também até 70oC por 1h, com atividade residual de 100% nas temperaturas de 55 a 60oC. A enzima purificada foi totalmente inibida por Ca2+, Cu2+, Hg2+ e 80% inibida por Fe2+ e Ag+. Mg2+ estimulou a atividade da poligalacturonase em 100%. Entre os substratos avaliados, a pectina de citrus (D.E. 92%) foi o que mais estimulou a ação da enzima. A afinidade por pectinas de alta esterificação e a ausência de atividade quando o ácido poligalacturônico foi usado como substrato, indica que a enzima purificada é uma polimetilgalacturonase. Os resultados obtidos pela cromatografia de papel demonstraram que a polimetilgalacturonase purificada possui o modo de clivagem endo/exo misto. O Km com pectina de citrus foi 10,78mg mL-1 e a Vmax foi 2379,04μmolmin-1mg-1. As possíveis aplicações desta enzima incluem composição de detergentes, processamento têxtil e de fibras de celulose, degradação ou modificação de material vegetal, aditivo... / Rhizomucor pusillus A 13.36 produced high levels of polygalacturonase (18.6U/g) in solid state fermentation in medium composed by cotton bran for 72 hours. This enzyme showed maximal activity at pH 5.5 and 55oC. Crude polygalacturonase was stable from pH 4.5 to 6.5, with remaining activity of 91,3% at pH 6.5, and stable until 70oC for 1h, with remaining activity of 80% at 55-60oC. This enzyme was purified by concentration and a combination of gel filtration and ion exchange chromatographic procedures. The molecular mass of the enzyme was 53.7kDa. Isoelectric focusing showed a single band with an isoelectric point (pI) of 3.8. The purified enzyme also showed maximum activity at pH 5.5 and 55oC. The purified enzyme was stable from pH 4.5 to 6.5, with remaining activity range of 80 to 100%, and stable until 70oC for 1h, with remaining activity of 100% at 55-60oC. The purified enzyme was totally inhibited by Ca2+, Cu2+, Hg2+ and 80% inhibited by Fe2+ and Ag+. Mg2+ increased poligalacturonase activity in 100%. Citrus pectin (D.E. 92%) was found to be the preferred substrate among the different substrates tested in the enzyme assay. The affinity for high pectin esterification and the lack of activity when polygalacturonic acid was used as substrate, indicates that the enzyme is a polymethylgalacturonase. The results obtained by paper chromatography showed that the purified polymethylgalacturonase has the mixed endo/exo mode of cleavage. The Km with citrus pectin was 10.78mg mL-1 and the Vmax was 2379.04μmolmin-1mg- 1. The possible applications of this enzyme include the composition of detergents, textile and cellulosic fiber processing, degradation or modification of plant material, animal feed additive and wine and juice processing.
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