Recovery and partial purification of penicillin G acylase from E. coli homogenate and B. megaterium culture medium using an expanded bed adsorption column

Pinotti, L.; Fonseca, Luís P.; Prazeres, D.M.F.; Rodrigues, Dasciana S.; Nucci, Edson Romano; Giordano, Raquel de Lima Camargo

doi:10.1016/j.bej.2008.11.006

Cited by 17 publications

(15 citation statements)

References 37 publications

(26 reference statements)

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“…The enzyme activity was determined by colorimetric PDAB method [23] and the cell concentration by optical density [24]. More details can be found in [9,15,24].…”

Section: Methodsmentioning

confidence: 99%

“…For production medium (g L -1 ), the same solution of amino acids (10.0), salt solution (0.5), cheese whey (19.6) and phenyl acetic acid (2.7) as inductor [9,15] have been used. All reactants used in the assays reported in this paper were taken from the same flasks.…”

Section: Culture Mediamentioning

confidence: 99%

“…This bacterium secretes the enzyme to the medium (one important aspect in this process), thus reducing downstream costs. Results concerning medium optimization for this process may be found elsewhere [7][8][9][10][11][12][13][14][15].…”

Section: Introductionmentioning

confidence: 99%

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Monitoring bioreactors using principal component analysis: production of penicillin G acylase as a case study

Nucci

Cruz

Giordano

2009

Bioprocess Biosyst Eng

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The complexity of biological processes often makes impractical the development of detailed, structured phenomenological models of the cultivation of microorganisms in bioreactors. In this context, data pre-treatment techniques are useful for bioprocess control and fault detection. Among them, principal component analysis (PCA) plays an important role. This work presents a case study of the application of this technique during real experiments, where the enzyme penicillin G acylase (PGA) was produced by Bacillus megaterium ATCC 14945. PGA hydrolyzes penicillin G to yield 6-aminopenicilanic acid (6-APA) and phenyl acetic acid. 6-APA is used to produce semi-synthetic beta-lactam antibiotics. A static PCA algorithm was implemented for on-line detection of deviations from the desired process behavior. The experiments were carried out in a 2-L bioreactor. Hotteling's T(2) was the discrimination criterion employed in this multivariable problem and the method showed a high sensibility for fault detection in all real cases that were studied.

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“…The enzyme activity was determined by colorimetric PDAB method [23] and the cell concentration by optical density [24]. More details can be found in [9,15,24].…”

Section: Methodsmentioning

confidence: 99%

Section: Culture Mediamentioning

confidence: 99%

See 1 more Smart Citation

Monitoring bioreactors using principal component analysis: production of penicillin G acylase as a case study

Nucci

Cruz

Giordano

2009

Bioprocess Biosyst Eng

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“…The EBA method is able to integrate primary purification, concentration, centrifugation, microfiltration, and other clarification prior steps into a single processing unit. Having these advantages, makes it possible to ensure high process efficiency, decrease the operation time, reduce the final costs of the product, and enhance the product quality [3][4][5][6][7][8][9]. It is desirable to know detailed information on the hydrodynamic characteristics and adsorption performance of nano/bioproducts in the EBA system to achieve better design, tighter control, scale-up, optimization, and finally greater efficiency of the EBA operation [10][11][12][13].…”

Section: Introductionmentioning

confidence: 98%

Hydrodynamic Characteristics and Adsorption Particularity of Nanobiological Feedstock Along the Bed Height in a Novel Chromatography Column

2012

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Expanded bed adsorption (EBA) is a practical method for the separation of nanoparticulates. In order to analysis the local hydrodynamic and adsorption behavior of nanoparticle (NP)-based biological feedstock, a modified Nano Biotechnology Group EBA column with a 26-mm inner diameter was used to withdraw liquid from different axial positions of the column. Fabricated egg albumin (EA) NPs with an average size of 70 nm were employed as a model system and viral size/charge mimic to assess the relationship between hydrodynamic and adsorption performance of NPs at the different column regions. The effects of influential factors, including flow velocity and initial concentration of NPs, on NP hydrodynamic behavior and adsorption kinetics along the bed height were investigated. NP hydrodynamic studies confirmed that non-uniform behavior dominated the system and a decreasing trend of liquid mixing/dispersion with increase of bed height was observed in this column. The results demonstrated an increase in the mixing/dispersion at certain bed heights with the increase in both the velocity and feed initial concentration. Breakthrough curves were measured at various column points to determine the adsorption performance [dynamic binding capacity (DBC) and yield] in different bed positions/zones. Yield and DBC of NPs were improved along the bed height, whereas liquid velocity had the opposite effect. Increasing the initial concentration of NPs enhanced only the DBC. Separation of EA NPs under optimal conditions was 87 %, which is an excellent result for a one-pass frontal chromatography method. KeywordsEBA chromatography Á Modified NBG column Á EA NPs Á Axial hydrodynamic behavior Á Axial adsorption performance List of symbols B o Bodenstein number C Exit concentration of NPs in bed effluent (kg m -3 ) C o Initial concentration of NPs (kg m -3 ) C 9Exit concentration that has left the 9-cm sample port and is measured at this port (kg m -3 ) C 12Exit concentration that has left the 12-cm sample port and is measured at this port (kg m -3 )Volume of feed loaded onto the column until C 9 reaches a certain degree of NP breakthrough (m 3 ) V f,12Volume of feed loaded onto the column until C 12 reaches a certain degree of NP breakthrough (m 3 ) V l Liquid volume above the particles in the sampling syringe (m 3 ) V L,0-9 Liquid volume present in the bed voidage for the 0-to 9-cm column zone (m 3 ) V L,0-12 Liquid volume present in the bed voidage for the 0-to 12-cm column zone (m 3 ) V s,0-9 Settled volume of adsorbent in the 0-to 9-cm column zone (m 3 ) V s,9-12 Settled volume of adsorbent in the 9-to 12-cm column zone (m 3 ) V o Volume of the particles in the sampling syringe (m 3 )Greek letters e Bed voidage e i Local bed voidage at ith layer of bed d Variance (s)

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“…On the other hand, EBA is a single-unit that combines several unit operation steps such as concentration, centrifugation, micro-filtration and other prior clarification steps. Therefore, this method has advantages over other conventional separation procedures such as decreasing the time of operation and total production costs and increasing product quality and process yield (Pinotti et al, 2009;Ng et al, 2007;da Silva Padilha et al, 2009;Vennapusa et al, 2009;. Analysis of adsorption performance is quite complex because of influential factors, namely fluid hydrodynamics, particle classification, and desired component transfer.…”

Section: Introductionmentioning

confidence: 99%

Investigation of hydrodynamic parameters in a novel expanded bed configuration: local axial dispersion characterization and an empirical correlation study

Taheri

Jahanshahi

Mosavian

et al. 2012

Braz. J. Chem. Eng.

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-Study of liquid behavior in an expanded bed adsorption (EBA) system is important for understanding, modeling and predicting nanobioproduct/biomolecule adsorption performance in such processes. In this work, in order to analyze the local axial dispersion parameters, simple custom NBG (Nano Biotechnology Group) expanded bed columns with 10 and 26 mm inner diameter were modified by insertion of sampling holes. Based on this configuration, the particles and liquid can be withdrawn directly from various axial positions of the columns. Streamline DEAE particles were used as solid phase in this work. The effects of factors such as liquid velocity, viscosity, settled bed height and column diameter on the hydrodynamic parameters were investigated. Local bed voidages in different axial bed positions were measured by a direct procedure within the column with 26 mm diameter. Increasing trend of voidage with velocity at a certain position of the bed and with bed height at a certain degree of expansion was observed. Residence time distribution (RTD) analysis at various bed points showed approximately uniform hydrodynamic behavior in the column with 10 mm diameter while a decreasing trend of mixing/dispersion along the bed height at a certain degree of expansion was seen in the column with 26 mm diameter. Also lower mixing/dispersion occured in the smaller diameter column. Finally, a combination of two empirical correlations proposed by Richardson-Zaki and Tong-Sun was successfully employed for identification of the bed voidage at various bed heights (RSSE=99.9%). Among the empirical correlations presented in the literatures for variation of the axial dispersion coefficient, the Yun correlation gave good agreement with our experimental data (RSSE=87%) in this column.

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Recovery and partial purification of penicillin G acylase from E. coli homogenate and B. megaterium culture medium using an expanded bed adsorption column

Cited by 17 publications

References 37 publications

Monitoring bioreactors using principal component analysis: production of penicillin G acylase as a case study

Monitoring bioreactors using principal component analysis: production of penicillin G acylase as a case study

Hydrodynamic Characteristics and Adsorption Particularity of Nanobiological Feedstock Along the Bed Height in a Novel Chromatography Column

Investigation of hydrodynamic parameters in a novel expanded bed configuration: local axial dispersion characterization and an empirical correlation study

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