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1.
Bioprocess Biosyst Eng ; 32(5): 625-32, 2009 Aug.
Article in English | MEDLINE | ID: mdl-19093135

ABSTRACT

The purification of clavulanic acid (CA), which is an important beta-lactam antibiotic produced by submerged cultivation of Streptomyces clavuligerus, was studied through the use of phosphate and polyethylene glycol-based aqueous two-phase systems. The parameters' effect on the yield and purification was evaluated through an experimental design and the preliminary results showed that the polyethylene molecular mass and tie-line length and phase volume ratio exerted the strongest effect on the yield and distribution coefficient in the range tested. In addition, the response surface methodology was used to optimize the distribution coefficient, yield, and purification factor. The optimal conditions of yield and purification factor are in the regions where polyethylene has a low molecular mass, pH close to the isoelectric point, and lower top phase volume. A 100% yield and a 1.5-fold purification factor are obtained when extracting CA by maximizing the conditions of an aqueous two-phase system.


Subject(s)
Bioreactors/microbiology , Clavulanic Acid/isolation & purification , Clavulanic Acid/metabolism , Models, Biological , Streptomyces/metabolism , Water/metabolism , Computer Simulation , Fermentation , Phase Transition , Water/chemistry
2.
Appl Biochem Biotechnol ; 105 -108: 705-13, 2003.
Article in English | MEDLINE | ID: mdl-12721408

ABSTRACT

This work compared the performance of three hydrophobic resins for the adsorption of ampicillin (AMP), D-phenylglycine (PG), D-phenylglycine methyl ester (PGME), and 6- aminopenicillanic acid (6-APA). The influence of pH on adsorption efficiencies was assessed in the range of 4.5-8.5, at 4 and 25 degrees C. The values at 4 degrees C were slightly higher than those at 25 degrees C. The adsorption efficiency of AMP and 6-APA decreased at higher pHs, for the three resins. An opposite behavior was found for PGME, and the pH did not affect PG adsorption efficiency. Isotherm models were fitted to experimental equilibrium data and the best models were discriminated.


Subject(s)
Ampicillin/metabolism , Penicillanic Acid/analogs & derivatives , Adsorption , Ampicillin/isolation & purification , Chromatography/methods , Hydrogen-Ion Concentration , Kinetics , Penicillanic Acid/isolation & purification , Thermodynamics
3.
Appl Biochem Biotechnol ; 105 -108: 867-79, 2003.
Article in English | MEDLINE | ID: mdl-12721424

ABSTRACT

Adsorption kinetics and equilibrium data of clavulanic acid, a beta-lactam antibiotic, on ion-exchange resin Amberlite IRA 400 were utilized to carry out the modeling and simulation of a continuous adsorption process. These simulations allowed the estimation of yield, concentration, and purification factors of the process utilizing the product final concentration. Experimental runs of this process were carried out using the conditions pointed out by simulation studies. Comparison of the experimental results and those calculated by the proposed model showed that the model could describe very well the main features of the continuous process.


Subject(s)
Anti-Bacterial Agents/isolation & purification , Clavulanic Acid/isolation & purification , Adsorption , Chromatography, Ion Exchange/methods , Computer Simulation , Kinetics , Models, Theoretical
4.
Appl Biochem Biotechnol ; 171(8): 2121-8, 2013 Dec.
Article in English | MEDLINE | ID: mdl-24026415

ABSTRACT

Cephamycin C (CepC) is a ß-lactam antibiotic that belongs to the cephalosporin class of drugs. This compound stands out from other cephalosporins for its greater resistance to ß-lactamases, which are enzymes produced by pathogenic microorganisms that present a major mechanism of bacterial resistance to ß-lactam antibiotics. Cephamycin C is produced by the bacterium Streptomyces clavuligerus. Knowledge about the stability of the compound under different values of pH is important for the development of the process of production, extraction, and purification aimed at obtaining higher yields. Therefore, the stability of cephamycin C under different pH levels (2.2, 6.0, 7.0, 7.6, and 8.7) at 20 °C was evaluated in this study. Ultrafiltered broth from batch fermentations of S. clavuligerus was used in the trials. The results indicated that cephamycin C is a more stable compound than other ß-lactam compounds such as penicillin and clavulanic acid. A higher degradation rate was observed at very acidic or basic pH levels, while this rate was lower at quasi-neutral pH levels. After 100 h of trial, the initial CepC showed 46 % degradation at pH 2.2, 71 % degradation at pH 8.7, and varied from 15 to 20 % at quasi-neutral pH levels.


Subject(s)
Cephamycins/chemistry , Cephamycins/isolation & purification , Streptomyces antibioticus/chemistry , beta-Lactam Resistance , Cephamycins/metabolism , Fermentation , Hydrogen-Ion Concentration , Kinetics , Streptomyces/chemistry , Streptomyces/metabolism , Streptomyces antibioticus/metabolism , beta-Lactamases/chemistry
5.
Appl Biochem Biotechnol ; 168(3): 651-71, 2012 Oct.
Article in English | MEDLINE | ID: mdl-22836750

ABSTRACT

This study evaluates the potential for using different effluents for simultaneous H(2) and CH(4) production in a two-stage batch fermentation process with mixed microflora. An appreciable amount of H(2) was produced from parboiled rice wastewater (23.9 mL g(-1) chemical oxygen demand [COD]) and vinasse (20.8 mL g(-1) COD), while other effluents supported CH(4) generation. The amount of CH(4) produced was minimum for sewage (46.3 mL g(-1) COD), followed by parboiled rice wastewater (115.5 mL g(-1) COD) and glycerol (180.1 mL g(-1) COD). The maximum amount of CH(4) was observed for vinasse (255.4 mL g(-1) COD). The total energy recovery from vinasse (10.4 kJ g(-1) COD) corresponded to the maximum COD reduction (74.7 %), followed by glycerol (70.38 %, 7.20 kJ g(-1) COD), parboiled rice wastewater (63.91 %, 4.92 kJ g(-1) COD), and sewage (51.11 %, 1.85 kJ g(-1) COD). The relatively high performance of vinasse in such comparisons could be attributed to the elevated concentrations of macronutrients contained in raw vinasse. The observations are based on kinetic parameters of H(2) and CH(4) production and global energy recovery of the process. These observations collectively suggest that organic-rich effluents can be deployed for energy recovery with sequential generation of H(2) and CH(4).


Subject(s)
Bacteria/metabolism , Hydrogen/metabolism , Methane/metabolism , Organic Chemicals/metabolism , Wastewater/microbiology , Biodegradation, Environmental , Bioreactors/microbiology , Fermentation , Hydrogen/chemistry , Kinetics , Methane/chemistry , Organic Chemicals/chemistry , Waste Disposal, Fluid , Wastewater/chemistry
6.
Appl Biochem Biotechnol ; 166(1): 208-21, 2012 Jan.
Article in English | MEDLINE | ID: mdl-22057909

ABSTRACT

A study was made for purification of cephamycin C from fermentation of Streptomyces clavuligerus. Initially, the culture broth was clarified by microfiltration and ultrafiltration, after which the resulting permeates were subjected to nonspecific adsorption and ion-exchange chromatography on resin columns. The antibiotic activity was measured by the biological method at each stage by assaying its activity against the Escherichia coli ESS, super sensitive to ß-lactam antibiotic. The purification processes were assessed in relation to the variables affecting each step. The purification efficiency by nonspecific adsorption was monitored by UV spectrophotometry, while the ion-exchange adsorption fractions were assessed by NMR spectroscopy. Some of the fractions obtained during purification were also analyzed by mass spectrometry (LC/MS and LC/MS/MS) to identify the cephamycin C molecule. These preliminary results proved the process feasibility.


Subject(s)
Anti-Infective Agents/chemistry , Anti-Infective Agents/isolation & purification , Cephamycins/chemistry , Cephamycins/isolation & purification , Streptomyces/chemistry , Anti-Infective Agents/pharmacology , Cephamycins/pharmacology , Chromatography, Ion Exchange , Escherichia coli/drug effects , Fermentation , Magnetic Resonance Spectroscopy , Mass Spectrometry , Recombinant Proteins , Spectrophotometry, Ultraviolet , beta-Lactams/chemistry
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