Bioluminescent sensor for naphthalene in air: Cell immobilization and evaluation with a dynamic standard atmosphere generator

A dynamic atmosphere generator with a naphthalene emission source has been constructed and used for the development and evaluation of a bioluminescence sensor based on the bacteria Pseudomonas fluorescens HK44 immobilized in 2% agar gel (101 cell mL(-1)) placed in sampling tubes. A steady naphthalene emission rate (around 7.3 nmol min(-1) at 27 degrees C and 7.4 mLmin(-1) of purified air) was obtained by covering the diffusion unit containing solid naphthalene with a PTFE filter membrane. The time elapsed from gelation of the agar matrix to analyte exposure (""maturation time"") was found relevant for the bioluminescence assays, being most favorable between 1.5 and 3 h. The maximum light emission, observed after 80 min, is dependent on the analyte concentration and the exposure time (evaluated between 5 and 20 min), but not on the flow rate of naphthalene in the sampling tube, over the range of 1.8-7.4 nmol min(-1). A good linear response was obtained between 50 and 260 nmol L-1 with a limit of detection estimated in 20 nmol L-1 far below the recommended threshold limit value for naphthalene in air. (c) 2008 Elsevier B.V. All rights reserved.

Cell immobilization improves isomaltulose production

The Laboratory of Industrial Biotechnology at the Biological Sciences Department in Sao Paulo State University, Brazil is working to improve the technologies involved with isomaltulose production. The study evaluated enzymatic reaction parameters with the goal of improving isomaltulose production which is grown with a medium of 1% cane molasses and 0.5% yeast extract thereby using calcium alginate, glutaraldehyde and polyethyleneimine. The best results were obtained using P. rubrum immobilized pellets in calcium alginate with 705 and 60% sucrose solution. The developed technology apparently allows the reuse of the cell-containing enzymes more times compared to conventional technologies, which ultimately results in decreased costs. The researchers are also involved in alcohol and biopolymer production and seeking interested industrial collaborators.

The whole-cell immobilization of D-hydantoinase-engineered Escherichia coli for D-CpHPG biosynthesis

Background: D-Hydroxyphenylglycine is considered to be an important chiral molecular building-block of antibiotic reagents such as pesticides, and β-lactam antibiotics. The process of its production is catalyzed by D-hydantoinase and D-carbamoylase in a two-step enzyme reaction. How to enhance the catalytic potential of the two enzymes is valuable for industrial application. In this investigation, an Escherichia coli strain genetically engineered with D-hydantoinase was immobilized by calcium alginate with certain adjuncts to evaluate the optimal condition for the biosynthesis of D-carbamoyl-p-hydroxyphenylglycine (D-CpHPG), the compound further be converted to D-hydroxyphenylglycine (D-HPG) by carbamoylase. Result: The optimal medium to produce D-CpHPG by whole-cell immobilization was a modified Luria-Bertani (LB) added with 3.0% (W/V) alginate, 1.5% (W/V) diatomite, 0.05% (W/V) CaCl2 and 1.00 mM MnCl2. The optimized diameter of immobilized beads for the whole-cell biosynthesis here was 2.60 mm. The maximized production rates of D-CpHPG were up to 76%, and the immobilized beads could be reused for 12 batches. Conclusions: This investigation not only provides an effective procedure for biological production of D-CpHPG, but gives an insight into the whole-cell immobilization technology. © 2016 Pontificia Universidad Católica de Valparaíso. Production and hosting by Elsevier B.V. All rights reserved.

Studies on immobilization of Bacteria

Cell immnhilizatinn technology in a rapidly expanding arna in the endeavour of microbial fnrmentatiwn.During the lnmt 15 years anveral prnceafinn have been developed and more are in developmental atage of approaching commercial utilizatinn.In the present programme it was planned to develop an optimized process for the innobilization of alpha amylase producing Bacillus polymyxa (CBTB 25) an isolate obtained from Cochin University campus primarily for the production of alpha-amylase.Optimal concentration of support material that attributaa stability and maximal activity to the immobilized cell beads was determined using different concentrations of sodium aliginate as support and estimation of amylase production.An overeall assessment of the data obtained for the various studies conducted denotes that immobilized cells synthesize alpha-amylase at comparable rates with free cells and produce reducing sugara at a higher level than free cells.Results indicated that both phosphate and citrate buffers could be used for disrupting the immobilized beads since they enforced maximal release of cells through leaching from the beads within one hour.On comparative analysis it was observed that immobilized cells could synthesize alpha amylase at similar levels with free cells of B.polymyxa.On Co-immobilization of B.Polymyxa with S.cerevisiae,the co-immobilizate beads could effeciently convert starch directly to ethanol with a yield of 14.8% at 1 : 2 ratio.

In vitro study on the cell adhesion ability of immobilized lactobacilli on natural supports

The aim of the present study was to investigate the effect of probiotic immobilization onto wheat grains, both wet and freeze dried, on the adhesion properties of the probiotic cells and make comparisons with wet and freeze dried free cells. Lactobacillus casei ATCC 393 and Lactobacillus plantarum NCIMB 8826 were used as model probiotic strains. The results showed satisfactory adhesion ability of free cells to a monolayer of Caco-2 cells (> 1000 CFU/100 Caco-2 cells for wet cells). Cell immobilization resulted in a significant decrease in adhesion, for both wet and freeze dried formulations, most likely because immobilized cells did not have direct access to the Caco-2 cells, but it still remained in adequate levels (> 100 CFU/100 Caco-2 cells for wet cells). No clear correlation could be observed between cell adhesion and the hydrophobicity of the bacterial cells, measured by the hexadecane adhesion assay. Most notably, immobilization enhanced the monolayer integrity of Caco-2 cells, demonstrated by a more than 2-fold increase in transepithelial electrical resistance (TEER) compared to free cells. SEM micrographs ascertained the adhesion of both immobilized and free cells to the brush border microvilli. Finally, the impact of the food matrix on the adhesion properties of probiotic bacteria and on the design of novel functional products is discussed.

Ferrous iron oxidation by foam immobilized Acidithiobacillus ferrooxidans: Experiments and modeling

Ferrous iron bio-oxidation by Acidithiobacillus ferrooxidans immobilized on polyurethane foam was investigated. Cells were immobilized on foams by placing them in a growth environment and fully bacterially activated polyurethane foams (BAPUFs) were prepared by serial subculturing in batches with partially bacterially activated foam (pBAPUFs). The dependence of foam density on cell immobilization process, the effect of pH and BAPUF loading on ferrous oxidation were studied to choose operating parameters for continuous operations. With an objective to have high cell densities both in foam and the liquid phase, pretreated foams of density 50 kg/m3 as cell support and ferrous oxidation at pH 1.5 to moderate the ferric precipitation were preferred. A novel basket-type bioreactor for continuous ferrous iron oxidation, which features a multiple effect of stirred tank in combination with recirculation, was designed and operated. The results were compared with that of a free cell and a sheet-type foam immobilized reactors. A fivefold increase in ferric iron productivity at 33.02 g/h/L of free volume in foam was achieved using basket-type bioreactor when compared to a free cell continuous system. A mathematical model for ferrous iron oxidation by Acidithiobacillus ferrooxidans cells immobilized on polyurethane foam was developed with cell growth in foam accounted by an effectiveness factor. The basic parameters of simulation were estimated using the experimental data on free cell growth as well as from cell attachment to foam under nongrowing conditions. The model predicted the phase of both oxidation of ferrous in shake flasks by pBAPUFs as well as by fully activated BAPUFs for different cell loadings in foam. Model for stirred tank basket bioreactor predicted within 5% both transient and steady state of the experiments closely for the simulated dilution rates. Bio-oxidation at high Fe2+ concentrations were simulated with experiments when substrate and product inhibition coefficients were factored into cell growth kinetics.

Influence of Major Operational Parameters on Aerobic Granulation for the Treatment of Wastewater

Effective solids-liquid separation is the basic concept of any wastewater treatment system. Biological treatment methods involve microorganisms for the treatment of wastewater. Conventional activated sludge process (ASP) poses the problem of poor settleability and hence require a large footprint. Biogranulation is an effective biotechnological process which can overcome the drawbacks of conventional ASP to a great extent. Aerobic granulation represents an innovative cell immobilization strategy in biological wastewater treatment. Aerobic granules are selfimmobilized microbial aggregates that are cultivated in sequencing batch reactors (SBRs). Aerobic granules have several advantages over conventional activated sludge flocs such as a dense and compact microbial structure, good settleability and high biomass retention. For cells in a culture to aggregate, a number of conditions have to be satisfied. Hence aerobic granulation is affected by many operating parameters. The organic loading rate (OLR) helps to enrich different bacterial species and to influence the size and settling ability of granules. Hence, OLR was argued as an influencing parameter by helping to enrich different bacterial species and to influence the size and settling ability of granules. Hydrodynamic shear force, caused by aeration and measured as superficial upflow air velocity (SUAV), has a strong influence and hence it is used to control the granulation process. Settling time (ST) and volume exchange ratio (VER) are also two key influencing factors, which can be considered as selection pressures responsible for aerobic granulation based on the concept of minimal settling velocity. Hence, these four parameters - OLR, SUAV, ST and VER- were selected as major influencing parametersfor the present study. Influence of these four parameters on aerobic granulation was investigated in this work

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.

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.

Bacterial nanocellulose for medical implants

Bacterial cellulose (BC) has established to be a remarkably versatile biomaterial and can be used in wide variety of applied scientific endeavours, especially for medical devices. In fact, biomedical devices recently have gained a significant amount of attention because of an increased interest in tissue-engineered products for both wound care and the regeneration of damaged or diseased organs. Due to its unique nanostructure and properties, microbial cellulose is a natural candidate for numerous medical and tissue-engineered applications. Hydrophilic bacterial cellulose fibers of an average diameter of 50 nm are produced by the bacterium Acetobacter xylinum, using a fermentation process. The microbial cellulose fiber has a high degree of crystallinity. Using direct nanomechanical measurement, determined that these fibers are very strong and when used in combination with other biocompatible materials, produce nanocomposites particularly suitable for use in human and veterinary medicine. Moreover, the nanostructure and morphological similarities with collagen make BC attractive for cell immobilization and cell support. The architecture of BC materials can be engineered over length scales ranging from nano to macro by controlling the biofabrication process. The chapter describes the fundamentals, purification and morphological investigation of bacterial cellulose. This chapter deals with the modification of microbial cellulose and how to increase the compatibility between cellulosic surfaces and a variety of plastic materials. Furthermore, provides deep knowledge of fascinating current and future applications of bacterial cellulose and their nanocomposites especially in the medical field, materials with properties closely mimic that of biological organs and tissues were described. © Springer-Verlag Berlin Heidelberg 2013.

FT-IR analysis of acid black dye biodegradation using saccharomyces cerevisiae immobilized with treated sugarcane bagasse

Textile industries use large amounts of water in dyeing processes and a wide variety of synthetic dyes. A small concentration of these dyes in the environment can generate highly visible pollution and changes in aquatic ecosystems. Adsorption, biosorption, and biodegradation are the most advantageous dye removal processes. Biodegradation occurs when enzymes produced by certain microorganisms are capable of breaking down the dye molecule. To increase the efficiency of these processes, cell immobilization enables the reuse of the immobilized cells and offers a high degree of mechanical strength, allowing metabolic processes to take place under adverse conditions. The aim of the present study was to investigate the use of Saccharomyces cerevisiae immobilized in activated sugarcane bagasse for the degradation of Acid Black 48 dye in aqueous solutions. For such, sugarcane bagasse was treated with polyethyleneimine (PEI). Concentrations of a 1 % S. cerevisiae suspension were evaluated to determine cell immobilization rates. Once immobilization was established, biodegradation assays for 240 h with free and immobilized yeast in PEI-treated sugarcane bagasse were evaluated by Fourier transform infrared spectrophotometry. The results indicated a probable change in the dye molecule and the possible formation of new metabolites. Thus, S. cerevisiae immobilized in sugarcane bagasse is very attractive for biodegradation processes in the treatment of textile effluents. © 2013 Springer Science+Business Media Dordrecht.

Seleção de um suporte sintético para imobilizar células do Botryospaheria rhodina e comparação da produção de lacase por células livres e imobilizadas

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Produção de etanol e levana por células de Zymomonas mobilis imobilizadas em alginato

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Produção de L-ácido lático a partir de células bacterianas imobilizadas

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Utilização de Saccharomyces cerevisiae imobilizada em bagaço de cana de açúcar para a biossorção e biodegradação do corante acid black 48

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)