Scielo RSS <![CDATA[Electronic Journal of Biotechnology]]> vol. 9 num. 4 lang. en <![CDATA[SciELO Logo]]> <![CDATA[<b>A new approach to chemical modification protocols of <i>Aspergillus niger </i>and sorption of lead ion by fungal species</b>]]> Biomasses of five fungi species (Aspergillus niger, Penicillium austurianum, Saccharomyces cerevisiae, Mucor arcindloides and Trichoderma reesi were evaluated for their uptake of lead ion from aqueous solution using batch systems. Both dead and live fungal biomasses were comparatively studied for their adsorption efficiencies. The effect of pH as one of the primary factors that influences sorption efficiency of metal ions in solution was also studied. Modification of the fungal biomass with the least sorption capacity was carried out using a four-step procedure. Three different modifying agents under the optimised experimental conditions were used. The percentage uptake of lead ion by fungi species ranged from: Aspergillus niger: 6.71-64.95% and 66.91-95.27%; Penicillium austurianum: 44.47-98.85% and 75.57-94.21%; Saccharomyces cerevisiae: 52.61-88.68% and 61.20-89.95%; Mucor arcindloides: 83.78-93.13% and 62.91-97.65% and Trichoderma reesi: 52.52-80.70% and 35.31-88.13% for dead and live biomass respectively. The influence of pH on metal uptake was evaluated at pH of 2, 4, 6 and 7. A regular pattern of sorption efficiency among the dead fungi species with respect to pH was observed and the % adsorption decreased in the order pH 7 > 2 > 6 > 4 with the exception of Mucor (pH 7 > 2 > 4 > 6). Modified biomass of Aspergillus niger with oxalic acid, malic acid and ethylenediamine tetraacetic acid (EDTA) recorded 92.84%, 48.11% and 39.83% uptake of Pb respectively which correspond to 69.65%, 41.23% and 29.25% increase when compared to 28.18% of the unmodified biomass. These quantitative adsorptions demonstrate the potential application of modified biomass for the removal of Pb ion from aqueous solution. <![CDATA[<B><I>Agrobacterium</I> <I>rhizogenes </I>mediated genetic transformation resulting in hairy root formation is enhanced by ultrasonication and acetosyringone treatment</B>]]> The phytopathogenic bacteria Agrobacterium rhizogenes genetically transforms plants by transferring a portion of the resident Ri plasmid, the T-DNA to the plant. Plant species differ in their temporal competence for transformation. But various physical and chemical methods are found to enhance transformation frequency. Agrobacterium rhizogenes mediated transformation efficiency was assessed under the influence of sonication, calcium treatment, acetosyringone and macerating enzymes in suitable combinations in Nicotiana tabacum as a model system. Manual wounding resulted in 21% transformation frequency. Where as sonication resulted in 2.2 fold increase, followed by sonication with CaCl2 treatment resulted in 2.5 fold increase and sonication with acetosyringone treatment resulted in 4.1 fold increase in transformation frequency. However, sonication with macerating enzyme treatment resulted in 1.5 to 5.25-fold decrease in transformation frequency. Micro wounding through sonication followed by acetosyringone treatment enhanced transformation frequency substantially. The results of this study may be very useful in genetic manipulation of plants by Agrobacterium rhizogenes mediated gene delivery to higher plants, which are recalcitrant to A. tumefaciens mediated genetic manipulation. <![CDATA[<b>Effect of glutaraldehyde biocide on laboratory-scale rotating biological contactors and biocide efficacy</b>]]> The effect of glutaraldehyde, a commercial biocide widely used in paper and pulp industry, on the performance of laboratory-scale rotating biological contactors (RBCs) as well as biocide efficacy was studied. Biofilms were established on the RBCs and then exposed to 0 - 180 ppm glutaraldehyde at a dilution rate of 1.60 h-1. The results showed that the biofilms became acclimated to glutaraldehyde and eventually could degrade it. Acclimation to the biocide took longer at the higher biocide concentrations. The degree of biocide degradation and chemical oxygen demand (COD) removal depended on acclimation period, the presence of other organic matters and the amount of mineral salts available. Glutaraldehyde at up to 80 ppm had no effect on treatment efficiency and populations of biofilms and planktonic phase of the system whereas glutaraldehyde at 180 ppm caused a progressive decline in all measured values. However, no glutaraldehyde concentration used in the study was sufficiently high to kill microorganisms in the RBC system. The presence of biofilm provided additional resistance to glutaraldehyde to bacteria because the biocide had to penetrate through biofilm to reach bacteria. The increased resistance of bacteria to glutaraldehyde due to acclimation should be considered in biocide applications. <![CDATA[Effect of sulphate concentration and sulphide desorption on the combined removal of organic matter and sulphate from wastewaters using expanded granular sludge bed (EGSB) reactors<B> </B>]]> During the application of anaerobic processes to high sulphate concentration wastewaters, operational problems are expected due to the occurrence of sulphate reduction. Sulphide production reduces effluent quality and may produce inhibition. The application of Expanded Granular Sludge Bed (EGSB) reactors for the combined removal of organic matter and sulphate was studied at different COD/sulphate and 3 values of pH. During the EGSB reactor operation, most of the sulphide remains in the liquid phase reducing effluent quality. The inclusion of a desorption column in the recirculation of the EGSB reactor promotes mass transfer to the gas phase, reducing the sulphide concentration in the liquid phase, significantly decreasing the chemical oxygen demand of the effluent. <![CDATA[<b>Improvement of myrosinase activity of <i>Aspergillus </i>sp. NR4617 by chemical mutagenesis</b>]]> A myrosinase (thioglucoside glucohydrolase or thioglucosidase, EC producing fungus, Aspergillus sp. NR4617, was newly isolated from decayed soil sample obtained in Thailand and was subjected to single exposure to two chemical mutagens, ethyl methanesulfonate (EMS) and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). Its myrosinase production was selected on low cost medium prepared from mustard seed cake (Brassica juncea). Studies of production and stability of the enzyme showed that EMS mutagenesis increased myrosinase activity. Aspergillus sp. NR4617E1 produced myrosinase 1.90 U ml-1 at 36 hrs of the cultivation equivalent to 171% of the enzyme production in wild-type. The stability studies revealed that myrosinase from the mutant strains retained activity similar to wild-type at 30&ordm;C. Aspergillus sp. NR4617E1 degraded 10 mM of glucosinolate completely in 36 hrs. Enhanced myrosinase production and high yields of products (allylisothiocyanate) demonstrated that this mutant could be a new found candidate for feed detoxification and industrial allylisothiocyanate production. <![CDATA[<b>Modulation of unsaturated fatty acids content in algae <i>Spirulina</i><i> platensis </i>and <i>Chlorella minutissima</i> in response to herbicide SAN 9785</b>]]> The accumulation of polyunsaturated fatty acids by algae Spirulina platensis and Chlorella minutissima was studied. Response of these organisms to the substituted pyridazinone, SAN 9785, an inhibitor of the long chain fatty acid desaturase, indicated that fatty acid synthesis and their desaturation were regulated differently in these organisms. While the pool of palmitic acid, the precursor for the unsaturated C18 fatty acids, was stringently maintained in the green alga C. minutissima, in the cyanobacterium S. platensis the level of palmitic acid was liberally maintained in spite of the enhanced accumulation of unsaturated C18 fatty acids. <![CDATA[<b>Optimization of culture conditions for exopolysaccharides production in <i>Rhizobium</i> sp. using the response surface method </b>]]> The combined effects of the processing parameters for exopolysaccharides production by Rhizobium sp. was studied using the experimental design and response surface methodology. The experiments were carried out using a fermenter with 20 L capacity, as the reactor. All processing parameters were online monitored. The temperature [(30 &plusmn; 1)&ordm;C] and pH value (7.0 &plusmn; 0.1) were kept constant throughout the experimental time. As statistical tools, a complete 2³ factorial planning with central point and response surface were used to study the interactions among three relevant variables of the fermentation process: calcium carbonate concentration, aeration and agitation. The processing parameters setup for reaching a maximum response for exopolysaccharides production was obtained when applying the highest values for calcium carbonate concentration (1.1 g/L), aeration (1.3 vvm) and agitation (800 rpm). In addition, the combination of these optimum processing parameters yielded Y P/S (g/g) = 0.35. <![CDATA[<B>Production and stability studies of the bioemulsifier obtained from a new strain of<I> Candida glabrata</I> UCP 1002</B>]]> Evaluation of both tenso-active and emulsifying activities indicated that a biosurfactant was produced by the newly isolated and promising strain Candida glabrata isolated from mangrove sediments. The extracellular water-soluble emulsifying agent was isolated and identified as a heteropolymer. The maximum of bioemulsifier production was observed when the strain was grown on soluble and insoluble substrates cotton seed oil plus glucose, reaching values of 10.0 g/l after 144 hrs at 200 rpm. The cell-free culture broth containing the examined agent lowered the surface tension of the medium to 31 mN/m. Stable and compact emulsions with emulsifying activity of 75% of cotton seed oil were detected. The emulsification capacity remained practically unaltered within a wide pH (2-12), temperature (4-80ºC) ranges and under NaCl concentrations up to 10%. <![CDATA[<B>Production of <I>Rhodotorula glutinis</B></I>: <B>a yeast that secretes <FONT FACE=Symbol>a</FONT>-L-arabinofuranosidase</B>]]> Rhodotorula glutinis is a yeast that secretes the enzyme alpha-L-arabinofuranosidase (E.C. into the culture medium and thus has an interesting biotechnological potential. To determine improved culture conditions of this organism, different factors of the culture media were evaluated such as the use of peptone as nitrogen source, salts composition, pH and growth temperature. Likewise, beet molasses and beet cosette were tested as industrial carbon sources to induce the production of the enzyme and how they influence the yeast growth. Based on these studies a culture medium is proposed for growth of this yeast in a continuous system. By assaying different dilution rates an average specific activity for the enzyme of 82.4 U/mg of protein was obtained. <![CDATA[<b>Oxygen and temperature effect on continuous citric acid secretion in <i>Candida oleophila</i></b>]]> The influence of air saturation and temperature on continuous citric acid secretion was studied in chemostat cultures of Candida oleophila ATCC 20177 (var.). Simultaneous measurements of intra- and extracellular concentration of glucose, citric and isocitric acid confirmed the involvement of a specific active transport system in citrate secretion, favouring citric acid over isocitrate. An optimum air oxygen saturation of 20% and temperature of 30-31&ordm;C were determined for the continuous citric acid secretion. The highest values of citric acid concentration (98 g/L), citrate to isocitrate ratio (33.3:1), volumetric citric productivity (1.8 g/(L x h)), and specific citric acid productivity (0.1 g/(g x h)), were reached at 20% air saturation at a residence time of 54 hrs by the experiment's lowest biomass of 18 g/L. The highest isocitic acid volumetric productivity (55.6 mg/(L x h)) and specific productivity (0.99 mg/(g x h)) were identified at 50%, instead. The fastest citrate excretion rate of the generic product of 0.046 g/(g*h) was found at 30-31&ordm;C. A concentration ratio between extra- and intracellular concentration of citrate of up to 9 was identified. The highest extra-/intracellular ratio of citrate and lowest intracellular concentrations of glucose, citric and isocitric acid were determined at optimum air saturation as a consequence of active citrate export. <![CDATA[<b>Safe use of genetically modified lactic acid bacteria in food</b>: <b>Bridging the gap between consumers, green groups, and industry</b>]]> Within the European Union (EU), the use of genetically modified organisms (GMOs) in food production is not widely applied and accepted. In contrast to the United States of America, the current EU legislation limits the introduction of functional foods derived from GMOs that may bring a clear benefit to the consumer. Genetically modified lactic acid bacteria (GM-LAB) can be considered as a different class of GMOs, and the European Union is preparing regulations for the risk assessment of genetically modified microorganisms. Since these procedures are not yet implemented, the current risk assessment procedure is shared for GMOs derived from micro organisms, plants, or animals. At present, the use of organisms in food production that have uncontrolled genetic alterations made through random mutagenesis, is permitted, while similar applications with organisms that have controlled genetic alterations are not allowed. The current paper reviews the opportunities that genetically modified lactic acid bacteria may offer the food industry and the consumer. An objective risk profile is described for the use of GM-LAB in food production. To enhance the introduction of functional foods with proven health claims it is proposed to adapt the current safety assessment procedures for (GM)-LAB and suggestions are made for the related cost accountability. A qualified presumption of safety as proposed by SANCO (<a href="#36">EU SANCO 2003</a>), based on taxonomy and on the history of safe use of LAB applied in food, could in the near future be applied to any kind of LAB or GM-LAB provided that a series of modern profiling methods are used to verify the absence of unintended effects of altered LAB that may cause harm to the health of the consumer. <![CDATA[<b>The lift pool method for isolation of cDNA clones from lambda phage libraries </b>]]> A PCR based strategy was developed to screen a Xenopus oocyte &lambda;gt10 cDNA library. The PCR-based lift pool (LP) method follows the same two tiered strategy as conventional screening of phage libraries by filter hybridization. Two rounds of plating, one at high density to detect the clone, and one at low density to purify the clone to homogeneity, are performed. In the first round, lysates from high density plates, termed plate pools (PP), serve as template for PCR. In the second round, phage particles adhering to plaque lifts of low density plates are washed off nitrocellulose filters to create LPs, which are used as template for PCR. The integrity of the plaques on the low-density plates is preserved. Once a positive LP has been identified, plaques on the corresponding plate are screened individually by PCR. Using isoform specific primer pairs for Xenopus myosin 7a and myosin 1d, two lambda clones were isolated. Subsequent DNA sequence analysis confirmed their identities as myosin isoforms (GenBank accession numbers: DQ100353 and AF540952). This method offers a time saving, cost-effective alternative to other hierarchical pooling strategies for the repeated screening by PCR of an arrayed lambda phage library. <![CDATA[<b>Cotton genetic diversity study by AFLP markers</b>]]> Amplified fragmentlength polymorphism (AFLP) markers have been used to ascertain the intensity of inherent diversity and relatedness in cotton (Gossypium spp.) plants. The effectiveness of this method to distinguish inter and intra specific difference in cotton could be handy in cultivar recognition and in marker assisted parental selection tool for plant breeders. Twenty cotton cultivars belonging to Gossypium hirsutum L., and G. arborium L. from the Pakistan and US origin were used for AFLP based genetic diversity estimates. The objective of this study was to assess the level of genetic variation among some cotton cultivars belonging to the old and new world species of cotton. Four EcoRI-MseI primer-pair combinations were used forthe AFLP analysis. The AFLP data assigned the genotypes into groups that corresponded with their origin and lineage relationships and showed a narrow genetic base among these cultivars.