RESUMEN
Exopolysaccharide (EPS) secretion by Sclerotium rolfsii ATCC 201126 in submerged cultures, already identified as high-osmolarity responsive, was assessed by reducing C-source without compromising EPS yields. A designed medium with 80 g sucrose L-1 (MOPT80) was tested at 3 L-bioreactor scale at different temperature, agitation, aeration and pH (uncontrolled vs. controlled) values. Optimal operative conditions (200 rpm, 28 °C, 0.5 vvm and initial pH -pHi- 4.5) were validated, as well as the possibility to work at pHi 5.5 to reduce biomass production. Purified EPSs produced in MOPT80 at optimal and other valid operative conditions exhibited refined grade (<1 % proteins and ash, 3-4 % reducing sugars, 87-99 % total sugars). EPS purity, MW and rheological parameters led to discourage pH controlled at 4.5. Relatively constant MW (6-8 × 106 Da) and outstanding viscosifying ability were found. Polyphasic EPS analysis (titre, purity, macromolecular features and rheological fitness) would support to properly select production conditions.
Asunto(s)
Basidiomycota/crecimiento & desarrollo , Reactores Biológicos , Glucanos/metabolismo , Basidiomycota/metabolismo , Biomasa , Reactores Biológicos/economía , Medios de Cultivo/química , Concentración de Iones de Hidrógeno , Reología , TemperaturaRESUMEN
BACKGROUND: Food residuals (FR) were anaerobically biotransformed to produce biogases (e.g. methane and hydrogen), and different pre-treatment conditions, including particle size, oil content, pH and salt content, were controlled in this study. The bio-solids of a municipal solid waste (MSW) from a wastewater treatment plant were added to assess its effect on anaerobic transformation efficiency and gas yields. RESULTS: The breaking of FR and the application of MSW were effective in enhancing the transformation efficiency and yield of biogases. The energy transfer efficiency value of the combined FRs used in this study was probably 23%. However, it can be very cost effective to apply arbitrary proportions to treat two types of FR in the anaerobic digestion tank of a wastewater treatment plant. It was also found that the alkalinity and pH value were two major parameters that controlled the success of the transformation. About 0.16-0.17 kg of alkalinity was needed during the anaerobic digestion of 1 kg dry FR, but this requirement was decreased by the treatment applying MSW. Olive oil had higher reducing rates when used as a substitute for heat-oxidized oil to study the effect of oil content on methylation. CONCLUSION: The conditions for anaerobic digestion established in this study were practical for the digestion of FR in wastewater treatment plants in Taiwan. However, we nonetheless found that it was cost effective to use arbitrary proportions for both types of FR and integrate the anaerobic digestion process used in wastewater treatment plants. © 2020 Society of Chemical Industry.
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Biocombustibles/análisis , Hidrógeno/análisis , Metano/análisis , Residuos Sólidos/análisis , Administración de Residuos/métodos , Anaerobiosis , Bacterias/metabolismo , Biocombustibles/microbiología , Reactores Biológicos/economía , Reactores Biológicos/microbiología , Biotransformación , Industria de Alimentos/economía , Hidrógeno/metabolismo , Residuos Industriales/análisis , Metano/metabolismo , Taiwán , Administración de Residuos/economíaRESUMEN
Biopharmaceutical protein production using transgenic plant cell bioreactor processes offers advantages over microbial and mammalian cell culture platforms in its ability to produce complex biologics with simple chemically defined media and reduced biosafety concerns. A disadvantage of plant cells from a traditional batch bioprocessing perspective is their slow growth rate which has motivated us to develop semicontinuous and/or perfusion processes. Although the economic benefits of plant cell culture bioprocesses are often mentioned in the literature, to our knowledge no rigorous technoeconomic models or analyses have been published. Here we present technoeconomic models in SuperPro Designer® for the large-scale production of recombinant butyrylcholinesterase (BChE), a prophylactic/therapeutic bioscavenger against organophosphate nerve agent poisoning, in inducible transgenic rice cell suspension cultures. The base facility designed to produce 25 kg BChE per year utilizing two-stage semicontinuous bioreactor operation manufactures a single 400 mg dose of BChE for $263. Semicontinuous operation scenarios result in 4-11% reduction over traditional two-stage batch operation scenarios. In addition to providing a simulation tool that will be useful to the plant-made pharmaceutical community, the model also provides a computational framework that can be used for other semicontinuous or batch bioreactor-based processes.
Asunto(s)
Productos Biológicos/economía , Reactores Biológicos/economía , Simulación por Computador/normas , Oryza/genética , Perfusión/métodos , Células Vegetales/metabolismo , Transgenes , Productos Biológicos/uso terapéutico , Reactores Biológicos/normas , Técnicas de Cultivo de Célula , Medios de Cultivo , Oryza/metabolismoRESUMEN
Process intensification has shown great potential to increase productivity and reduce costs in biomanufacturing. This case study describes the evolution of a manufacturing process from a conventional processing scheme at 1000-L scale (Process A, n = 5) to intensified processing schemes at both 1000-L (Process B, n = 8) and 2000-L scales (Process C, n = 3) for the production of a monoclonal antibody by a Chinese hamster ovary cell line. For the upstream part of the process, we implemented an intensified seed culture scheme to enhance cell densities at the seed culture step (N-1) prior to the production bioreactor (N) by using either enriched N-1 seed culture medium for Process B or by operating the N-1 step in perfusion mode for Process C. The increased final cell densities at the N-1 step allowed for much higher inoculation densities in the production bioreactor operated in fed-batch mode and substantially increased titers by 4-fold from Process A to B and 8-fold from Process A to C, while maintaining comparable final product quality. Multiple changes were made to intensify the downstream process to accommodate the increased titers. New high-capacity resins were implemented for the Protein A and anion exchange chromatography (AEX) steps, and the cation exchange chromatography (CEX) step was changed from bind-elute to flow-through mode for the streamlined Process B. Multi-column chromatography was developed for Protein A capture, and an integrated AEX-CEX pool-less polishing steps allowed semi-continuous Process C with increased productivity as well as reductions in resin requirements, buffer consumption, and processing times. A cost-of-goods analysis on consumables showed 6.7-10.1 fold cost reduction from the conventional Process A to the intensified Process C. The hybrid-intensified process described here is easy to implement in manufacturing and lays a good foundation to develop a fully continuous manufacturing with even higher productivity in the future.
Asunto(s)
Anticuerpos Monoclonales/metabolismo , Reactores Biológicos/economía , Biotecnología/organización & administración , Animales , Biotecnología/economía , Células CHO , Técnicas de Cultivo de Célula , Proliferación Celular , Costos y Análisis de Costo , Cricetinae , Cricetulus , Eficiencia , Humanos , Invenciones , Modelos EconómicosRESUMEN
Lignin is an abundant and heterogeneous waste byproduct of the cellulosic industry, which has the potential of being transformed into valuable biochemicals via microbial fermentation. In this study, we applied a fast-pyrolysis process using softwood lignin resulting in a two-phase bio-oil containing monomeric and oligomeric aromatics without syringol. We demonstrated that an additional hydrodeoxygenation step within the process leads to an enhanced thermochemical conversion of guaiacol into catechol and phenol. After steam bath distillation, Pseudomonas putida KT2440-BN6 achieved a percent yield of cis, cis-muconic acid of up to 95 mol% from catechol derived from the aqueous phase. We next established a downstream process for purifying cis, cis-muconic acid (39.9 g/L) produced in a 42.5 L fermenter using glucose and benzoate as carbon substrates. On the basis of the obtained values for each unit operation of the empirical processes, we next performed a limited life cycle and cost analysis of an integrated biotechnological and chemical process for producing adipic acid and then compared it with the conventional petrochemical route. The simulated scenarios estimate that by attaining a mixture of catechol, phenol, cresol, and guaiacol (1:0.34:0.18:0, mol ratio), a titer of 62.5 (g/L) cis, cis-muconic acid in the bioreactor, and a controlled cooling of pyrolysis gases to concentrate monomeric aromatics in the aqueous phase, the bio-based route results in a reduction of CO2 -eq emission by 58% and energy demand by 23% with a contribution margin for the aqueous phase of up to 88.05 euro/ton. We conclude that the bio-based production of adipic acid from softwood lignins brings environmental benefits over the petrochemical procedure and is cost-effective at an industrial scale. Further research is essential to achieve the proposed cis, cis-muconic acid yield from true lignin-derived aromatics using whole-cell biocatalysts.
Asunto(s)
Adipatos/metabolismo , Reactores Biológicos , Lignina/metabolismo , Reactores Biológicos/economía , Reactores Biológicos/microbiología , Fermentación , Fenoles/metabolismo , Pseudomonas putida/metabolismo , Pirólisis , Ácido Sórbico/análogos & derivados , Ácido Sórbico/metabolismoRESUMEN
Omega-3 fatty acids, and specifically docosahexaenoic acid (DHA), are important and essential nutrients for human health. Thraustochytrids are recognised as commercial strains for nutraceuticals production, they are group of marine oleaginous microorganisms capable of co-synthesis of DHA and other valuable carotenoids in their cellular compartment. The present study sought to optimize DHA and squalene production by the thraustochytrid Schizochytrium limacinum SR21. The highest biomass yield (0.46 g/gsubstrate) and lipid productivity (0.239 g/gsubstrate) were observed with 60 g/L of glucose, following cultivation in a bioreactor, with the DHA content to be 67.76% w/wtotal lipids. To reduce costs, cheaper feedstocks and simultaneous production of various value-added products for pharmaceutical or energy use should be attempted. To this end, we replaced pure glucose with organosolv-pretreated spruce hydrolysate and assessed the simultaneous production of DHA and squalene from S. limacinum SR21. After the 72 h of cultivation period in bioreactor, the maximum DHA content was observed to 66.72% w/wtotal lipids that was corresponded to 10.15 g/L of DHA concentration. While the highest DHA productivity was 3.38 ± 0.27 g/L/d and squalene reached a total of 933.72 ± 6.53 mg/L (16.34 ± 1.81 mg/gCDW). In summary, we show that the co-production of DHA and squalene makes S. limacinum SR21 appropriate strain for commercial-scale production of nutraceuticals.
Asunto(s)
Reactores Biológicos/microbiología , Ácidos Docosahexaenoicos/biosíntesis , Microbiología Industrial/métodos , Escualeno/metabolismo , Estramenopilos/metabolismo , Biocombustibles/economía , Biomasa , Reactores Biológicos/economía , Análisis Costo-Beneficio , Suplementos Dietéticos , Ácidos Docosahexaenoicos/aislamiento & purificación , Bosques , Microbiología Industrial/economía , Escualeno/aislamiento & purificación , Desarrollo SostenibleRESUMEN
The aim of this work was to investigate the technical as well as the economic feasibility of producing 2-ethyl hexyl oleate (2-EHO), a non-phthalate plasticizer in a solvent free medium. The esterification reaction between oleic acid and 2-ethyl hexyl alcohol was carried out in a packed bed reactor (PBR) using Candida antarctica lipase B (Novozym 435; Novozymes; Copenhagen-Denmark) as biocatalyst. RSM was employed to optimize the esterification reaction conditions. The optimum reaction conditions were found to be flow rate of 1.5 mL/min, No. of cycles of 12 and molar ratio of 4:1 2-ethyl hexanol to oleic acid. The maximum experimental and predicated conversions were found to be 95.8% and 95.61% respectively. Formation of 2-EHO was approved by FTIR, 1HNMR and 13CNMR. From the economic prospective, PBR was capable of producing 2-EHO with a purity of more than 94% over 480 h without remarkable reduction of enzyme activity. This revealed an economic production of 2-EHO at a yield of 2 tons kg-1 lipase. The manufacturing cost was found to be $ 1.88 /kg 2-EHO, this contributed to a profit of about 30% compared to the commercial price of 2-EHO. Such results approve the technical and economic feasibility for this sustainable method in esters production.
Asunto(s)
Reactores Biológicos , Éteres de Etila/síntesis química , Plastificantes/síntesis química , Reactores Biológicos/economía , Costos y Análisis de Costo , Enzimas Inmovilizadas , Esterificación , Éteres de Etila/química , Proteínas Fúngicas , Lipasa/química , Ácido Oléico/química , Plastificantes/químicaRESUMEN
The present study deals with optimizing, producing, characterizing, application and techno- economic analysis of oxidative enzymes [Laccase (Lac), manganese peroxidase (MnP), and lignin peroxidase (LiP)] from Aspergillus iizukae EAN605 in submerged fermentation process using pumpkin peels as a production substrate. The best operating parameters for producing Lac, MnP and LiP (6.15, 2.58 and 127.99 U mg-1 respectively) were recorded with 20â¯g 100â¯mL-1 of substrate, 4.6â¯mL 100â¯mL-1 of inoculum size at pH 5.5 after 10 days. The crude enzyme exhibited high stability at pH (3-9) and temperatures (20-60⯰C). Km (Michaelis-Menten) of Lac, MnP and LiP crude enzyme was 2.25, 1.79 and 0.72â¯mM respectively. The decolourization of Remazol Brilliant Blue R by the crude enzyme was 84.84 %. The techno-economic analysis was assessed for a production unit with an annual operating time for enzymatic production and application is 7920â¯h/year and 100 m3 of the capacity. The process would produce 27,000â¯cm3 of crude enzyme with a price of USD 0.107 per cm3 compared to USD 1 per cm3 of the current commercial enzyme. The findings indicated that pumpkin peels have potential as a production substrate for oxidative enzymes from A. iizukae EAN605 and is economically feasible.
Asunto(s)
Aspergillus/enzimología , Reactores Biológicos/microbiología , Cucurbita/química , Lacasa/aislamiento & purificación , Peroxidasas/aislamiento & purificación , Reactores Biológicos/economía , Celulosa/química , Análisis Costo-Beneficio , Fermentación , Especificidad por SustratoRESUMEN
As antibiotic-resistant bacterial pathogens become an ever-increasing concern, antimicrobial peptides (AMPs) have grown increasingly attractive as alternatives. Potentially, plants could be used as cost-effective AMP bioreactors; however, reported heterologous AMP expression is much lower in plants than in E. coli expression systems and often results in plant cytotoxicity, even for AMPs fused to carrier proteins. This suggests that there may be a physical characteristic of the previously described heterologous AMPs which impedes efficient expression in plants. Using a meta-analysis of protein databases, this study has determined that native plant AMPs were significantly less cationic than AMPs native to other taxa. To apply this finding to plant expression, the transient expression of 10 different heterologous AMPs, ranging in charge from +7 to -5, was tested in the tobacco, Nicotiana benthamiana. Elastin-like polypeptide (ELP) was used as the carrier protein for AMP expression. ELP fusion allowed for a simple, cost-effective temperature shift purification. Using this system, all five anionic AMPs expressed well, with two at unusually high levels (375 and 563⯵g/gfw). Furthermore, antimicrobial activity against Staphylococcus epidermidis was an order of magnitude greater (average minimum inhibitory concentration MIC of 0.26µM) than that typically seen for AMPs expressed in E. coli systems and was associated with the uncleaved fusion peptide. In summary, this study describes a means of expressing AMP fusions in plants in high yield, purified by a simple temperature-shift protocol, resulting in a fusion peptide with high antimicrobial activity and without the need for a peptide cleavage step.
Asunto(s)
Antibacterianos/farmacología , Péptidos Catiónicos Antimicrobianos/farmacología , Reactores Biológicos/economía , Nicotiana/química , Staphylococcus epidermidis/efectos de los fármacos , Antibacterianos/química , Antibacterianos/metabolismo , Péptidos Catiónicos Antimicrobianos/química , Péptidos Catiónicos Antimicrobianos/metabolismo , Bases de Datos de Proteínas , Relación Dosis-Respuesta a Droga , Pruebas de Sensibilidad Microbiana , Temperatura , Nicotiana/metabolismoRESUMEN
Biological sulfur reduction is an attractive sulfidogenic technology for the treatment of organics-deficient metal-laden wastewater, because it theoretically reduces the electron donor consumption by 75%, compared to sulfate reduction. However, reducing the external organic substrate dosage may lower the sulfur reduction rate. Supplying with a more biodegradable organic substrate could possibly enhance sulfidogenic activity but also increase the chemical cost. Therefore, the sulfide production performance of a sulfur-reducing bioreactor feeding with varied levels of organic supply, and different types of organic substrates were investigated. The results showed that high-rate sulfide production (12.30â¯mgâ¯S/L/h) in a sulfur-reducing bioreactor can be achieved at the minimal dosage of organic substrate as low as 39â¯mgâ¯C/L of organic carbon in the influent. Changing the type of organic substrate posed a significant effect on the sulfidogenic activity in the sulfur-reducing bioreactor. Sodium acetate was found to be the optimal substrate to achieve the highest sulfide production rate (28.20â¯mgâ¯S/L/h) by sulfur-reducing bacteria (S0RB), followed by ethanol, methanol, glycerol, pyruvic acid, acetic acid, glucose, sucrose, malic acid, sodium formate, formic acid, N-propanol, N-butanol, lactic acid, sodium lactate, propionic acid and sodium propionate (2.87â¯mgâ¯S/L/h as the lowest rate). However, the cost-effectiveness analysis showed that glucose was the most cost-effective organic substrate to realize the sulfur reduction process in high sulfide production rate (20.13â¯mgâ¯S/L/h) and low chemical cost (5.94â¯kgâ¯S/$). The utilization pathway of the different organic substrates in the sulfur-reducing bioreactor was also discussed.
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Reactores Biológicos/microbiología , Compuestos Orgánicos/análisis , Bacterias Reductoras del Azufre/metabolismo , Azufre/metabolismo , Purificación del Agua/métodos , Reactores Biológicos/economía , Análisis Costo-Beneficio , Metales/metabolismo , Compuestos Orgánicos/economía , Compuestos Orgánicos/metabolismo , Oxidación-Reducción , Sulfatos/metabolismo , Sulfuros/metabolismo , Eliminación de Residuos Líquidos/métodos , Aguas Residuales/microbiología , Purificación del Agua/economíaRESUMEN
Traditionally, aeration units, used as a polishing stage after anaerobic digestion (AD) of wastes, are operated at ambient temperature. Yet, when effluent quality is the main design criterion, raising the temperature of the aeration stage can be justified by improved removal efficiencies. In this study, an anaerobic-aerobic sequential system (AASS) was operated to co-digest raw wastewater and food waste. The aerobic compartment was tested under psychrophilic and mesophilic temperatures. At the design loading rate of 2 gVS L-1 d-1, the anaerobic digester achieved removal efficiencies of 85 ± 2% of volatile solids (VS), 84 ± 3% of total chemical oxygen demand (CODT) and a biogas yield of 1,035 ± 30 mL gVSfed-1 (50% methane). The aerobic reactor achieved additional removal of 8% CODT and 7 % VS. By raising the temperature of the aerobic reactor to the mesophilic range, COD and solids concentrations of the effluent dropped to approximately half their values. This was accompanied by an increase in nitrification (from 68% to 91%) and denitrification (from 10% to 16%). The energy analysis showed that total energy consumption slightly increases (from 0.45 to 0.49 kWh kgCODfed-1) by raising the temperature of the aerobic reactor to mesophilic range. A preliminary evaluation of the sludge disposal cost, revealed a saving increase of 5-6% under mesophilic operation with respect to psychrophilic conditions. Implications: In order to cope with the globally increasing constraints on the disposal of urban wastes, efficient post-processing of effluents becomes a crucial requirement for the anaerobic digestion industry. In this context, the submitted manuscript shows that the quality of the effluent, of an anaerobic digester, treating food waste with raw wastewater, can be substantially improved by optimizing the aerobic polishing stage. Raising the temperature of the aerobic reactor to the mesophilic range resulted in a drop of solids and COD concentrations to approximately half their values. Equally important, the implications on operational costs were found to be favorable, compared to traditional psychrophilic aerobic post-treatment, when taking into consideration indirect sludge treatment costs and energy selling revenues.
Asunto(s)
Reactores Biológicos , Eliminación de Residuos Líquidos/métodos , Aerobiosis , Anaerobiosis , Reactores Biológicos/economía , Costos y Análisis de Costo , Alimentos , Temperatura , Eliminación de Residuos Líquidos/economía , Aguas ResidualesRESUMEN
In the current work, the attempt was made to apply best-fitted artificial neural network (ANN) architecture and the respective training process for predicting final titer of hepatitis B surface antigen (HBsAg), produced intracellularly by recombinant Pichia pastoris Mut+ in the commercial scale. For this purpose, in large-scale fed-batch fermentation, using methanol for HBsAg induction and cell growth, three parameters of average specific growth rate, biomass yield, and dry biomass concentration-in the definite integral form with respect to fermentation time-were selected as input vectors; the final concentration of HBsAg was selected for the ANN output. Used dataset consists of 38 runs from previous batches; feed-forward ANN 3:5:1 with training algorithm of backpropagation based on a Bayesian regularization was trained and tested with a high degree of accuracy. Implementing the verified ANN for predicting the HBsAg titer of the five new fermentation runs, excluded from the dataset, in the full-scale production, the coefficient of regression and root-mean-square error were found to be 0.969299 and 2.716774, respectively. These results suggest that this verified soft sensor could be an excellent alternative for the current relatively expensive and time-intensive analytical techniques such as enzyme-linked immunosorbent assay in the biopharmaceutical industry.
Asunto(s)
Reactores Biológicos , Fermentación , Antígenos de Superficie de la Hepatitis B/metabolismo , Redes Neurales de la Computación , Pichia/metabolismo , Reactores Biológicos/economía , Antígenos de Superficie de la Hepatitis B/química , Antígenos de Superficie de la Hepatitis B/economía , Pichia/química , Proteínas Recombinantes/química , Proteínas Recombinantes/economía , Proteínas Recombinantes/metabolismoRESUMEN
Accumulation of plastic pollution in aquatic ecosystems is the predictable result of high demand for plastic functionalities, optimized production with economies of scale, and recalcitrance. Strategies are needed for end-of-life conversion of recalcitrant plastics into useful feedstocks and for transition to materials that are biodegradable, non-bioaccumulative, and non-toxic. Promising alternatives are the polyhydroxyalkanoates (PHAs), a vast family of polymers amenable to decentralized production from renewable feedstocks. Establishment of a global-scale PHA-based industry will require identification of PHAs with tailored properties for use as 'drop-in' replacements for existing plastics; use of low-cost renewable/waste-derived feedstocks; high productivity cultures that may be genetically modified microorganisms or non-axenic mixed cultures maintained by selection pressures that favor high PHA-producing strains; and low-cost extraction/purification schemes.
Asunto(s)
Biotecnología/métodos , Plásticos/análisis , Biodegradación Ambiental , Reactores Biológicos/economía , Biotecnología/economía , Desarrollo SostenibleRESUMEN
Scalable production of kilobase single-stranded DNA (ssDNA) with sequence control has applications in therapeutics, gene synthesis and sequencing, scaffolded DNA origami, and archival DNA memory storage. Biological production of circular ssDNA (cssDNA) using M13 addresses these needs at low cost. However, one unmet goal is to minimize the essential protein coding regions of the exported DNA while maintaining its infectivity and production purity to produce sequences less than 3,000 nt in length, relevant to therapeutic and materials science applications. Toward this end, synthetic miniphage with inserts of custom sequence and size offers scalable, low-cost synthesis of cssDNA at milligram and higher scales. Here, we optimize growth conditions using an E. coli helper strain combined with a miniphage genome carrying only an f1 origin and a ß-lactamase-encoding (bla) antibiotic resistance gene, enabling isolation of pure cssDNA with a minimum sequence genomic length of 1,676 nt, without requiring additional purification from contaminating DNA. Low-cost scalability of isogenic, custom-length cssDNA is demonstrated for a sequence of 2,520 nt using a bioreactor, purified with low endotoxin levels (<5 E.U./ml). We apply these exonuclease-resistant cssDNAs to the self-assembly of wireframe DNA origami objects and to encode digital information on the miniphage genome for biological amplification.
Asunto(s)
Reactores Biológicos/virología , ADN de Cadena Simple/biosíntesis , Escherichia coli/metabolismo , Microbiología Industrial/métodos , Bacteriófago M13/genética , Reactores Biológicos/economía , ADN de Cadena Simple/aislamiento & purificación , Escherichia coli/genética , Escherichia coli/virología , Microbiología Industrial/economía , Nanotecnología/economía , Nanotecnología/métodos , Plásmidos/genéticaRESUMEN
Biodiesel production depends to a great extent on the use of cheap raw materials, since biodiesel itself is a mass product, not a high-value product. New processing methods, such as micro-flow continuous processing combined with enzymatic catalysis, open doors to the latter. As reported here, the window of opportunity in enzyme-catalyzed biodiesel production is the conversion of waste cooking oil. The main technological challenge for this is to obtain efficient immobilization of the lipase catalyst on beads. The beads can be filled into tubular reactors where designed packed-bed provide porous channels, forming micro-flow. It turns out, that in this way, the immobilization costs become the decisive economic factor. This paper reports a solution to that issue. The use of oil cake enables economic viability, which is not given by any of the commercial polymeric substrates used so far for enzyme immobilization. The costs of immobilization are mirrored in the earnings and cash flow of the new biotechnological process.
Asunto(s)
Biocombustibles , Reactores Biológicos , Biotecnología/economía , Grasas Insaturadas en la Dieta , Enzimas Inmovilizadas/metabolismo , Lipasa/metabolismo , Biocombustibles/análisis , Biocombustibles/economía , Reactores Biológicos/economía , Biotecnología/instrumentación , Biotecnología/métodos , Catálisis , Culinaria , Análisis Costo-Beneficio , Grasas Insaturadas en la Dieta/economía , Grasas Insaturadas en la Dieta/metabolismo , Enzimas Inmovilizadas/economía , Esterificación , Humanos , Dispositivos Laboratorio en un Chip/economía , Microtecnología/instrumentación , Microtecnología/métodos , Aceites de Plantas/química , Aceites de Plantas/metabolismoRESUMEN
This study focuses on the optimisation of 2,3-butanediol (BDO) production in fed-batch cultures carried out with the bacterial strain Enterobacter ludwigii using very high polarity (VHP) sugar from sugarcane mills. Various kLa values were evaluated using either complex or synthetic fermentation media demonstrating that the latter enhance BDO production efficiency with low by-product formation. The pH (6.3) and temperature (33.9⯰C) employed in fed-batch bioreactor cultures has been optimised via experimental design. Fed-batch cultures carried out at the optimum temperature and pH and varying kLa values resulted in BDO concentration, yield and productivity of 86.8â¯g/L, 0.37â¯g/g and 3.95â¯gâ¯L-1â¯h-1. Using this fermentation efficiency, the minimum selling price of BDO for annual production capacities of 10,000â¯t and 50,000â¯t was estimated at $3.12/kg and $2.67/kg, respectively, for a VHP cane sugar market price of $0.4/kg.
Asunto(s)
Reactores Biológicos , Butileno Glicoles/metabolismo , Saccharum/metabolismo , Técnicas de Cultivo Celular por Lotes , Reactores Biológicos/economía , Reactores Biológicos/microbiología , Fermentación , TemperaturaRESUMEN
Mesenchymal stromal cell (MSC) therapies have been pursued for a broad spectrum of indications but mixed reports on clinical efficacy have given rise to some degree of skepticism regarding the effectiveness of this approach. However, recent reports of successful clinical outcomes and regulatory approvals for graft-versus-host disease, Crohn's disease and critical limb ischemia have prompted a shift in this perspective. With hundreds of clinical trials involving MSCs currently underway and an increasing demand for large-scale manufacturing protocols, there is a critical need to develop standards that can be applied to processing methods and to establish consensus assays for both MSC processing control and MSC product release. Reference materials and validated, uniformly applied tests for quality control of MSC products are needed. Here, we review recent developments in MSC manufacturing technologies, release testing and potency assays. We conclude that, although MSCs hold considerable promise clinically, economies of scale have yet to be achieved although numerous bioreactor technologies for scalable production of MSCs exist. Additionally, rigorous disease-specific product testing and comprehensive understanding of mechanisms of action, which are linked to relevant process and product release potency assays, will be required to ensure that these therapies continue to be successful.
Asunto(s)
Técnicas de Cultivo Celular por Lotes/métodos , Bioensayo/métodos , Trasplante de Células Madre Mesenquimatosas/métodos , Células Madre Mesenquimatosas , Control de Calidad , Reactores Biológicos/clasificación , Reactores Biológicos/economía , Supervivencia Celular/fisiología , Enfermedad de Crohn/terapia , Endotoxinas/análisis , Enfermedad Injerto contra Huésped/terapia , Humanos , Mycoplasma , Resultado del TratamientoRESUMEN
The changing landscape of the biopharmaceutical market is driving a paradigm shift toward continuous manufacturing. To date, integrated continuous bioprocessing has not been realized as enabling technologies are nascent. In this work, a fully integrated continuous process is successfully demonstrated from pilot scale bioreactor to drug substance. Comparable product quality is observed between the continuous process and a 500 L fed-batch conventional process. The continuous process generated material at a rate of 1 kg of purified mAb every 4 days, achieving a 4.6-fold increase in productivity compared to the fed-batch process A plant throughput analysis using BioSolve software shows that a fed-batch facility with 4 × 12 500 L stainless steel bioreactors and purification train of the corresponding scale can be replaced by a continuous facility consisting of 5 × 2000 L single use bioreactors and smaller purification train, with a cost reduction of 15%.
Asunto(s)
Anticuerpos Monoclonales/biosíntesis , Anticuerpos Monoclonales/aislamiento & purificación , Técnicas de Cultivo Celular por Lotes/economía , Técnicas de Cultivo Celular por Lotes/métodos , Industria Farmacéutica/economía , Técnicas de Cultivo Celular por Lotes/instrumentación , Biofarmacia/economía , Biofarmacia/métodos , Reactores Biológicos/economía , Costos y Análisis de Costo , Modelos Teóricos , Programas Informáticos , Factores de Tiempo , Flujo de TrabajoRESUMEN
Over the last several years, continuous manufacturing of pharmaceuticals has evolved from bulk APIs and solid oral dosages into the more complex realm of biologics. The development of continuous downstream processing techniques has allowed biologics manufacturing to realize the benefits (e.g., improved economics, more consistent quality) that come with continuous processing. If relevant processing techniques and principles are selected, the opportunity arises to develop continuous manufacturing designs for additional pharmaceutical products including liposomal drug formulations. Liposome manufacturing has some inherent aspects that make it favorable for a continuous process. Other aspects such as formulation refinement, materials of construction, and aseptic processing need development, but present an achievable challenge. This paper reviews the current state of continuous manufacturing technology applicable to liposomal drug product manufacturing and an assessment of the challenges and potential of this application.
Asunto(s)
Productos Biológicos/aislamiento & purificación , Reactores Biológicos , Industria Farmacéutica/tendencias , Liposomas/aislamiento & purificación , Tecnología Farmacéutica/tendencias , Productos Biológicos/economía , Reactores Biológicos/economía , Reactores Biológicos/normas , Costos y Análisis de Costo , Industria Farmacéutica/economía , Industria Farmacéutica/normas , Liposomas/economía , Control de Calidad , Tecnología Farmacéutica/economía , Tecnología Farmacéutica/normas , Flujo de TrabajoRESUMEN
The biopharmaceutical industry is evolving in response to changing market conditions, including increasing competition and growing pressures to reduce costs. Single-use (SU) technologies and continuous bioprocessing have attracted attention as potential facilitators of cost-optimized manufacturing for monoclonal antibodies. While disposable bioprocessing has been adopted at many scales of manufacturing, continuous bioprocessing has yet to reach the same level of implementation. In this study, the cost of goods of Pall Life Science's integrated, continuous bioprocessing (ICB) platform is modeled, along with that of purification processes in stainless-steel and SU batch formats. All three models include costs associated with downstream processing only. Evaluation of the models across a broad range of clinical and commercial scenarios reveal that the cost savings gained by switching from stainless-steel to SU batch processing are often amplified by continuous operation. The continuous platform exhibits the lowest cost of goods across 78% of all scenarios modeled here, with the SU batch process having the lowest costs in the rest of the cases. The relative savings demonstrated by the continuous process are greatest at the highest feed titers and volumes. These findings indicate that existing and imminent continuous technologies and equipment can become key enablers for more cost effective manufacturing of biopharmaceuticals.