Studies on the respiration rate of free and immobilized cells of Cephalosporium acremonium in cephalosporin C production

Bioprocesses using filamentous fungi immobilized in inert supports present many advantages when compared to conventional free cell processes. However, assessment of the real advantages of the unconventional process demands a rigorous study of the limitations to diffusional mass transfer of the reagents, especially concerning oxygen. In this work, a comparative study was carried out on the cephalosporin C production process in defined medium containing glucose and sucrose as main carbon and energy sources, by free and immobilized cells of Cephalosporium acremonium ATCC 48272 in calcium alginate gel beads containing alumina. The effective diffusivity of oxygen through the gel beads and the effectiveness factors related to the respiration rate of the microorganism were determined experimentally. By applying Monod kinetics, the respiration kinetics parameters were experimentally determined in independent experiments in a complete production medium. The effectiveness factor experimental values presented good agreement with the theoretical values of the approximated zero-order effectiveness factor, considering the dead core model. Furthermore, experimental results obtained with immobilized cells in a 1.7-L tower bioreactor were compared with those obtained in 5-L conventional fermenter with free cells. It could be concluded that it is possible to attain rather high production rates working with relatively large diameter gel beads (ca. 2.5 mm) and sucrose consumption-based productivity was remarkably higher with immobilized cells, i.e., 0.33 gCPC/kg sucrose/h against 0.24 gCPC/kg sucrose/h in the aerated stirred tank bioreactor process. (C) 1999 John Wiley & Sons, Inc.

Acid Black 48 dye biosorption using Saccharomyces cerevisiae immobilized with treated sugarcane bagasse

The textile industry consumes large quantities of water and chemicals, especially in dyeing and finishing processes. Textile dye adsorption can be accomplished with natural or synthetic compounds. Cell immobilization using biomaterials allows the reduction of toxicity and mechanical resistance and opens spaces within the matrix for cell growth. The use of natural materials, such as sugarcane bagasse, is promising due to the low costs involved. The aim of the present study was to evaluate the use of sugarcane bagasse treated with either polyethyleneimine (PEI), NaOH or distilled water in the cell immobilization of Saccharomyces cerevisiae for textile dye removal. Three different adsorption tests were conducted: treated sugarcane bagasse alone, free yeast cells and bagasse-immobilized yeast cells. Yeast immobilization was 31.34% with PEI-treated bagasse, 8.56% with distilled water and 22.54% with NaOH. PEI-treated bagasse exhibited the best removal rates of the dye at all pH values studied (2.50, 4.50 and 6.50). The best Acid Black 48 adsorption rates were obtained with use of free yeast cells. At pH 2.50, 1 mg of free yeast cells was able to remove 5488.49 g of the dye. The lowest adsorption capacity rates were obtained using treated bagasse alone. However, the use of bagasse-immobilized cells increased adsorption efficiency from 20 to 40%. The use of immobilized cells in textile dye removal is very attractive due to adsorbed dye precipitation, which eliminates the industrial need for centrifugation processes. Dye adsorption using only yeast cells or sugarcane bagasse requires separation methods.

Electrocatalytic study of an electrode modified with Reactive Blue 4 dye covalently immobilized on amine-functionalized silica

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Speciation of lead in seawater and river water by using Saccharomyces cerevisiae immobilized in agarose gel as a binding agent in the diffusive gradients in thin films technique

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

On-line redox speciation analysis of antimony using L-proline immobilized on controlled pore glass and hydride generation inductively coupled plasma optical emission spectrometry for detection

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Use of Saccharomyces cerevisiae immobilized in agarose gel as a binding agent for diffusive gradients in thin films

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Stimulation of polygalacturonase production in an immobilized system by Aspergillus sp.: effect of pectin and glucose

The synthesis of polygalacturonases (PG) is known to be influenced by Aspergillus growth conditions, namely, environmental factors and pectin content in the cultivation medium containing a mixed carbon source. Optimal conditions were attained at a temperature of 30 A degrees C and an initial pH of 4.5. PG activity (3.29 and 2.48 U/mL) was determined after a two-day culture of Aspergillus sp. HC1 and Aspergillus sp. CC1, respectively, in a basic medium containing 2% citrus pectin as the sole carbon source. The addition of glucose (2% w/v) to the basic medium led to a 2-fold increase in PG production. However, enzyme synthesis was repressed when a higher concentration of glucose was used in the medium containing the mixed carbon source. Spores from the two fungi were immobilized in a 3% Ca-alginate system and the mechanical strength of the gel beads allowed the use of this process system 6-fold longer (288 h) than the free culture. In the Aspergillus sp. CC1 immobilized system, PG production increased nearly 10-fold in the medium with 2% glucose added (5.95 U/mL) in comparison to the medium without sugar (0.55 U/mL). The results demonstrate that a different response in activity was produced by free and entrapped spore systems. PG production remained approximately constant throughout the six 48 h cycles in the medium containing citrus pectin (2% w/v) as the sole carbon source.

Synthesis and characterization of ruthenium complexes immobilized on poly(4-vinylpyridine)

The [Ru(NH3)5(H2O)]2+ and trans-[Ru(NH3)4SO2(H2O)]2+ complexes ions were immobilized on poly(4-vinylpyridine) (4-PVP) through reactions in aqueous solutions. The stability of the imobilized complexes was checked in aqueous solution in the pH 2.0-8.0 range. The number of pyridinic nitrogens in the polymer 4-PVP is 2.80±0.05 mmol/g according to nitrogen elemental analysis. Potentiometric titration experiments showed that the accessible nitrogen, in aqueous medium, was 0.94±0.02 mmol/g with a p Ka value of 7.4±0.2. In addition, ruthenium and sulfate analysis has demonstrated that about 15% of the accessible nitrogen sites are able to coordinate to the metal centers. The characterization of the immobilized complexes was made through diffuse electronic and infrared spectroscopies and differential pulse and cyclic voltammetries. © 1993 Plenum Publishing Corporation.

Oxalate determination in urine using an immobilized enzyme on Sorghum vulgare seeds in a flow injection conductimetric system

A flow-injection (FI) method was developed for the determination of oxalate in urine. It was based on the use of oxalate oxidase (E.C. 1.2.3.4) immobilized on ground seeds of the BR-303 Sorghum vulgare variety. A reactor was filled with this activated material, and the samples (200 μL) containing oxalate were passed through it, carried by a deionized water flow. The carbon dioxide produced by the enzyme reaction permeated through a microporous PTFE membrane, and was received in a water acceptor stream, promoting conductivity changes proportional to the oxalate concentration in the sample. The results obtained showed a useful linear range from 0.05 to 0.50 mmol dm-3. The proposed method, when compared with the Sigma enzymatic procedure, showed good correlation (Y = 0.006(±0.016) + 0.98(±0.019)X; r = 0.9995, Y = conductivity in μS, and X = concentration in mmol dm-3), selectivity, and sensitivity. The new immobilization approach promotes greater stability, allowing oxalate determination for 6 months. About 13 determinations can be performed per hour. The precision of the proposed method is about ± 3.2 % (r.s.d).

A novel potentiometric naproxenate ion sensor immobilized in a graphite matrix for determination of naproxen in pharmaceuticals

The characteristics, performance, and application of an electrode, namely, Pt|Hg|Hg 2(NAP) 2| Graphite, where NAP stands for naproxenate ion, are described. This electrode responds to NAP with sensivity of (58.1± 0.9) mV decade -1 over the range 5.0 × 10 -5 - 1.0 × 10 -2 mol L -1 at pH 6.0-9.0 and a detection limit of 3.9 × 10 -5 mol L -1. The electrode is easily constructed at a relatively low cost with fast response time (within 10-35 s) and can be used for a period of 6 months without any considerable divergence in potentials. The proposed sensor displayed good selectivity for naproxen in the presence of several substances, especially concerning carboxylate and inorganic anions. It was used for the direct assay of naproxen in commercial tablets by means of the standard additions method. The analytical results obtained by using this electrode are in good agreement with those given by the United States Pharmacopeia procedure. ©2006 Sociedade Brasileira de Química.

Solubilization of iron phosphate by free or immobilized spores and pellets of Aspergillus niger

The production of pellets by Aspergillus niger and the FePO 4 (FeP) solubilization through the use of free or immobilized spores and pellets were studied. The media Sabouraud, MS-FeP, MS-K 2HPO 4, Czapek and Malt were used for pellets yield. Enhanced growth and pellets production were found in the Sabouraud and MS-FeP than those produced by the other media. Pellets produced in Sabouraud were larger in size and formed well-defined spheres than those produced in MS-FeP. Pellets amounts varying from 2 to 8 mg mL, -1 were inoculated in the MS-FeP medium. The greatest quantity of solubilized FeP was found when 6 mg of pellets mL, -1 were used. While the free spores were the worst form used for FeP solubilization in culture medium, free pellets allowed for the production of the greatest quantities of soluble phosphate, even after repeated use of the pellets. In the soil, pellets solubilized similar quantities of FeP compared to the immobilized A. niger spores and can be used with advantage since they are easily produced. © 2010 Academic Journals Inc.

Characterization of a tannase from Emericela nidulans immobilized on ionic and covalent supports for propyl gallate synthesis

The extracellular tannase from Emericela nidulans was immobilized on different ionic and covalent supports. The derivatives obtained using DEAE-Sepharose and Q-Sepharose were thermally stable from 60 to 75 °C, with a half life (t50) >24 h at 80 °C at pH 5. 0. The glyoxyl-agarose and amino-glyoxyl derivatives showed a thermal stability which was lower than that observed for ionic supports. However, when the stability to pH was considered, the derivatives obtained from covalent supports were more stable than those obtained from ionic supports. DEAE-Sepharose and Q-Sepharose derivatives as well as the free enzyme were stable in 30 and 50 % (v/v) 1-propanol. The CNBr-agarose derivative catalyzed complete tannic acid hydrolysis, whereas the Q-Sepharose derivative catalyzed the transesterification reaction to produce propyl gallate (88 % recovery), which is an important antioxidant. © 2012 Springer Science+Business Media Dordrecht.

Acid dye biodegradation using saccharomyces cerevisiae immobilized with polyethyleneimine-treated sugarcane bagasse

Chemical reagents used by the textile industry are very diverse in their composition, ranging from inorganic compounds to polymeric compounds. Strong color is the most notable characteristic of textile effluents, and a large number of processes have been employed for color removal. In recent years, attention has been directed toward various natural solid materials that are able to remove pollutants from contaminated water at low cost, such as sugarcane bagasse. Cell immobilization has emerged as an alternative that offers many advantages in the biodegradation process, including the reuse of immobilized cells and high mechanical strength, which enables metabolic processes to occur under adverse conditions of pH, sterility, and agitation. Support treatment also increases the number of charges on the surface, thereby facilitating cell immobilization processes through adsorption and ionic bonds. Polyethyleneimine (PEI) is a polycationic compound known to have a positive effect on enzyme activity and stability. The aim of the present study was to investigate a low-cost alternative for the biodegradation and bioremediation of textile dyes, analyzing Saccharomyces cerevisiae immobilization in activated bagasse for the promotion of Acid Black 48 dye biodegradation in an aqueous solution. A 1 % concentration of a S. cerevisiae suspension was evaluated to determine cell immobilization rates. Once immobilization was established, biodegradation assays with free and immobilized yeast in PEI-treated sugarcane bagasse were evaluated for 240 h using UV-vis spectrophotometry. The analysis revealed significant relative absorbance values, indicating the occurrence of biodegradation in both treatments. Therefore, S. cerevisiae immobilized in sugarcane bagasse is very attractive for use in biodegradation processes for the treatment of textile effluents. © 2012 Springer Science+Business Media Dordrecht.

Production of xylo-oligosaccharides by immobilized-stabilized derivatives of endo-xylanase from Streptomyces halstedii

An endoxylanase from Streptomyces halstedii was stabilized by multipoint covalent immobilization on glyoxyl-agarose supports. The immobilized enzyme derivatives preserved 65% of the catalytic activity corresponding to the one of soluble enzyme that had been immobilized. These immobilized derivatives were 200 times more stable 200 times more stable than the one-point covalently immobilized derivative in experiments involving thermal inactivation at 60 °C. The activity and stability of the immobilized enzyme was higher at pH 5.0 than at pH 7.0. The optimal temperature for xylan hydrolysis was 10 °C higher for the stabilized derivative than for the non-stabilized derivative. On the other hand, the highest loading capacity of activated 10% agarose gels was 75 mg of enzyme per mL of support. To prevent diffusional limitations, low loaded derivatives (containing 0.2 mg of enzyme per mL of support) were used to study the hydrolysis of xylan at high concentration (close to 1% (w/v)). 80% of the reducing sugars were released after 3 h at 55 °C. After 80% of enzymatic hydrolysis, a mixture of small xylo-oligosaccharides was obtained (from xylobiose to xylohexose) with a high percentage of xylobiose and minimal amounts of xylose. The immobilized-stabilized derivatives were used for 10 reaction cycles with no loss of catalytic activity. © 2013 Elsevier Ltd. All rights reserved.