Cultivo de células de hibridoma murino en el bioreactor-fermentador LAMBDA MINIFOR. 2017:
Mouse Hybridoma Cell Culture in a Protein-Free Medium Using a Bio-Mimicking Fish-Tail Disc Stirred Bioreactor. Valdés R, Aragón H, González M, Hernández D, Geada D, Goitizolo D et al. BioProcessing Journal, Volume 16, Issue: 1, Spring 2017. Pages: 51–64. https://doi.org/10.12665/J161.Valdes. Center for Genetic Engineering and Biotechnology. Cuba. Palabras claves: Cultivo de hibridoma, CB.Hep-1 , Biomimicking fish-tail, bioreactor.
Resumen: Because the Lambda MINIFOR bioreactor provides good mixing of cell culture, nutrients, and gases without any damaging hydrodynamic forces by using a bio-mimicking “fish-tail“ disc stirrer, it can be successfully applied for the cultivation of bacteria and yeast, insect, plant, and mammalian cells. However, reports on its application in mouse hybridoma cell culture using protein-free media is non-existent in the scientific literature. Therefore, this study describes preliminary findings of the Lambda MINIFOR bioreactor suitability in mouse hybridoma cell culture and antibody production using the SP2/O-Ag14-CB.Hep-1 mouse hybridoma cell and the PFHM-II protein-free medium as models. [showhide type="link" more_text="Leer más" less_text="Ocultar"]Results verified 2.45 × 106 viable cells/mL as the highest cell concentration, 86% as maximum cell viability, 0.0156/h as the exponential growth rate, 44 h as cell population doubling time, a stable phenotype measured by limiting dilution after 2.5 months, no antibiotic and antifoam requirements, 71.4% of IgG SDS-PAGE purity in the cell culture harvested supernatant, 38.68 ± 22.29 μg/mL, 39.23 ± 10.66 pg/cell/day, up to 99.5% of purity (sample measured by SDS-PAGE and SE-HPLC) after an IgG capture step based on protein A-Sepharose, a low pH incubation, and size-exclusion chromatography, no molecule aggregation, specificity for the CKTCTT epitope (located in the HBsAg “a” determinant), an IgG affinity constant equal to 1.11 × 1010 M-1, and < 78 pg mouse DNA/mg of IgG. In conclusion, this study corroborated a cumulative CB.Hep-1 mAb production of 1.77 g/15 days and validated the usefulness of the Lambda MINIFOR bioreactor in mouse hybridoma cell culture in protein-free media for research applications...
1. Evaluación del efecto de la carencia de azúcares bajo varias condiciones de oxigenación O2 en células aisladas de vegetales y cultivadas en el Lambda Minifor. 2017:
Metabolic profiling reveals a coordinated response of isolated lamb's (Valerianella locusta, L.) lettuce cells to sugar starvation and low oxygen stress. Baiye Mfortaw Mbong Victor, Jerry Ampofo-Asiama, Maarten Hertog, Annemie H Geeraerd and Bart M. Nicolai. Postharvest Biology and Technology. Volume 126, April 2017, Pages: 23-33. Department of Biochemistry, University of Cape Coast, Ghana. Palabras claves: Células de Lechuga Lamb (Valerianella locusta L.); Carencia de azúcares; Estrés a baja concentración de O2; Respuesta metabólica; Perfil de metabolitos; Marcaje 13C.
Resumen: Sugar starvation is a common phenomenon occurring in most leafy vegetables after harvest and storage. Additionally, leafy vegetables are subjected to low O2 stress when stored in modified atmosphere conditions. In this study, the metabolism of isolated lamb’s lettuce cells was studied upon sugar starvation under O2 stress conditions, using 13C labelled glucose. Fast depletions of the soluble sugars were observed, being more pronounced under aerobic conditions than under low O2 stress conditions. Sugar starvation under aerobic conditions resulted in increased levels and decreased 13C label incorporation of TCA cycle intermediates and amino and fatty acids originating from glycolytic and TCA cycle pathways, compared to starving cells incubated under low O2stress. [showhide type="link" more_text="Leer más" less_text="Ocultar"]On incubation under low O2 stress a switch in metabolism from aerobic to fermentation metabolism was observed. Under low O2 stress conditions, increased levels and 13C label incorporated in hexose phosphates, pyruvate, lactate, GABA, alanine, together with increased levels of acetaldehyde, ethanol and ethyl acetate was observed indicating fermentative metabolism was triggered.
2. Robusta producción de etanol celulósico a partir de bagazo de la caña de azúcar con Saccharomyces cerevisiae ATCC 20602 en el bioreactor LAMBDA MINIFOR bajo condiciones aeróbicas y anaeróbicas con medición controlada del potencial Redox: 2016.
Catalytic conversion of sugarcane bagasse to cellulosic ethanol: TiO2 coupled nanocellulose as an effective hydrolysis enhancer. Jabasingh, S. Anuradha, D. Lalith, M. Arun Prabhu, Abubekker Yimam, and Taye Zewdu. Carbohydrate Polymers (2016.Carbohydrate Polymers, Volume 136, 20 January 2016, Pages 700-709). Addis Ababa Institute of Technology, Ethiopia; Sathyabama University, India. Palabras claves: Etanol celulósico, bagazo, dióxido de titanio, nanocelulosa, celulasa de Saccharomyces cerevisiae
Resumen: The present study deals with the production of cellulosic ethanol from bagasse using the synthesized TiO2 coupled nanocellulose (NC-TiO2) as catalyst. Aspergillus nidulans AJSU04 cellulase was used for the hydrolysis of bagasse. NC-TiO2 at various concentrations was added to bagasse in order to enhance the yield of reducing sugars. Complex interaction between cellulase, bagasse, NC-TiO2 and the reaction environment is thoroughly studied. A mathematical model was developed to describe the hydrolysis reaction. Ethanol production from enzymatically hydrolyzed sugarcane bagasse catalyzed with NC-TiO2 was carried out using Saccharomyces cerevisiae ATCC 20602. The glucose release rates and ethanol concentrations were determined. [showhide type="link" more_text="Leer más" less_text="Ocultar"]Ethanol produced was found to be strongly dependent on pretreatment given, hydrolysis and fermentation conditions. The study confirmed the promising accessibility of NC-TiO2, for enhanced glucose production rates and improved ethanol yield.
3. Expresión de la proteína S. pyogenes Cas9 usando el bioreactor MINIFOR con vaso de 3L controlado por una computadora en modo de operación batch seguida de una alimentación exponencial: 2015.
Homology-directed repair in rodent zygotes using Cas9 and TALEN engineered proteins.Ménoret, Séverine, Anne De Cian, Laurent Tesson, Séverine Remy, Claire Usal, Jean-Baptiste Boulé, Charlotte Boix et al. Scientific reports5 (2015). 2015; 7 Oct 2015. Nature ublishing Group. INSERM UMR 1064-ITUN; CNRS UMS3556 Nantes; CNRS UMR7196; Sorbonne Universities; University Pierre & Marie Curie; France.
Resumen: The generation of genetically-modified organisms has been revolutionized by the development of new genome editing technologies based on the use of gene-specific nucleases, such as meganucleases, ZFNs, TALENs and CRISPRs-Cas9 systems. The most rapid and cost-effective way to generate genetically-modified animals is by microinjection of the nucleic acids encoding gene-specific nucleases into zygotes. However, the efficiency of the procedure can still be improved. In this work we aim to increase the efficiency of CRISPRs-Cas9 and TALENs homology-directed repair by using TALENs and Cas9 proteins, instead of mRNA, microinjected into rat and mouse zygotes along with long or short donor DNAs. [showhide type="link" more_text="Leer más" less_text="Ocultar"]We observed that Cas9 protein was more efficient at homology-directed repair than mRNA, while TALEN protein was less efficient than mRNA at inducing homology-directed repair. Our results indicate that the use of Cas9 protein could represent a simple and practical methodological alternative to Cas9 mRNA in the generation of genetically-modified rats and mice as well as probably some other mammals.
4. Fermentación de un microorganismo modificado por ingeniería genética en el bioreactor de escala de laboratorio MINIFOR, para lograr una conversión eficiente de lactosa a etanol: 2015.
"Methods for genetic optimization of biocatalysts for biofuel production from dairy waste through synthetic biology."Pasotti, L., S. Zucca, M. Casanova, N. Politi, I. Massaiu, G. Mazzini, G. Micoli, C. Calvio, M. G. Cusella De Angelis, and P. Magni. In Engineering in Medicine and Biology Society (EMBC), 2015 37th Annual International Conference of the IEEE, pp. 953-956. IEEE, 2015. Department of Electrical, Computer & Biomedical Engineering and Interdepartmental Research Centre for Tissue Engineering, University of Pavia, Italy. Palabras claves: conversión lactosa a etanol; optimización de microorganismos, biología sintética, proteíanas del trigo, permeato, disposición de desechos de polutantes; optimización genética; producción de energía verde, producción de biofuel, proceso de producción de queso, desecho diario, biocatalizador.
Resumen: Whey is an abundant by-product of cheese production process and it is considered a special waste due to its high nutritional load and hypertrophic potential. Technologies for whey valorization are available. They can convert such waste into high-value products, like whey proteins. However, the remaining liquid (called permeate) is still considered as a polluting waste due to its high lactose concentration. The alcoholic fermentation of lactose into ethanol will simultaneously achieve two important goals: safe disposal of a pollutant waste and green energy production. This methodology paper illustrates the workflow carried out to design and realize an optimized microorganism that can efficiently perform the lactose-to-ethanol conversion, engineered via synthetic biology experimental and computational approaches.
5. Un modelo de biofilme celular de flujo con 6 especies se desarrolló y se cultivaron las bacterias en el fermentador LAMBDA MINIFOR para evaluar el desarrollo del biofilme bajo condiciones de fuerzas de corte y flujo: 2015.
The use of in vitro model systems to study dental biofilms associated with caries: a short review. Salli, Krista M., and Arthur C. Ouwehand.Journal of oral Microbiology 7 (2015). J Oral Microbiol. 2015; 7: 10.3402/jom.v7.26149. DuPont Nutrition and Health, Kantvik Active Nutrition, Finland. Palabras claves: caries dental, cultivo en batch o lote, cultivo continuo, boca artificial, flujo celular y microcosmos.
Resumen: A dental biofilm forms a distinct environment where microorganisms live in a matrix of extracellular polysaccharides. The biofilm favors certain bacteria and creates a habitat that functions differently compared to planktonic bacteria. Reproducible model systems which help to address various questions related to biofilm formation, the process of caries development, and its prevention are needed and are continuously developed. Recent research using both batch culture, continuous culture and flow cells in caries biofilm formation is presented. The development of new techniques and equipment has led to a deeper understanding of how caries biofilms function. Biofilm models have also been used in the development of materials inhibiting secondary caries. This short review summarizes available models to study these questions.
6. Cuantificación de las proteínas ribosomales (RPs) a partir de células de levaduras cultivadas en el MINIFOR y células madres embrionarias de ratón (ESC) para estudiar la estequeometría del core o núcleo de las RPs: 2015.
Differential stoichiometry among core ribosomal proteins, arXiv:1406.0399 Nikolai Slavov, Stefan Semrau, Edoardo Airoldi, Bogdan Budnik, Alexander van Oudenaarden, , Jun 2015. Cell Reports 13 , 865–873 November 3, 2015. Harvard University, USA; Broad Institute of MIT and Harvard, USA and Hubrecht Institute, Netherlands . Palabras claves: Levaduras, células madres embrionarias (ESC), Proteínas ribosomales, RP, ribosomas, mRNA, espectrometría de masa, modificaciones posttraduccionales.
Resumen: Understanding the regulation and structure of ribosomes is essential to understanding protein synthesis and its deregulation in disease. While ribosomes are believed to have a fixed stoichiometry among their core ribosomal proteins (RPs), some experiments suggest a more variable composition. Testing such variability requires direct and precise quantification of RPs. We used mass-spectrometry to directly quantify RPs across monosomes and polysomes of mouse embryonic stem cells (ESC) and budding yeast. Our data show that the stoichiometry among core RPs in wild-type yeast cells and ESC depends both on the growth conditions and on the number of ribosomes bound per mRNA. Furthermore, we find that the fitness of cells with a deleted RP-gene is inversely proportional to the enrichment of the corresponding RP in polysomes. Together, our findings support the existence of ribosomes with distinct protein composition and physiological function.
7. Crecimiento de cultivos de levaduras (DBY12007) en el MINIFOR en estado estacionario para estudiar la glicólisis aerobia y el flujo de energía: 2014.
Constant Growth Rate Can Be Supported by Decreasing Energy Flux and Increasing Aerobic Glycolysis. Nikolai Slavov, Bogdan A. Budnik, David Schwab, Edoardo M. Airoldi, and Alexander van Oudenaarden, Cell Reports 7, 705–714, May 8, 2014 Massachusetts Institute of Technology, USA; Harvard University, USA; Hubrecht Institute, Netherlands and Princeton University, USA. Palabras claves: Levaduras, glicólisis aerobia, crecimiento exponencial, consumo de O2, producción de CO2, aminoácidos, mRNAs, proteínas, modificaciones posttraduccionales, sensibilidad al estrés, coeficiente respiratorio (RQ).
Resumen: Fermenting glucose in the presence of enough oxygen to support respiration, known as aerobic glycolysis, is believed to maximize growth rate. We observed increasing aerobic glycolysis during exponential growth, suggesting additional physiological roles for aerobic glycolysis. We investigated such roles in yeast batch cultures by quantifying O2 consumption, CO2 production, amino acids, mRNAs, proteins, posttranslational modifications, and stress sensitivity in the course of nine doublings at constant rate. During this course, the cells support a constant biomass-production rate with decreasing rates of respiration and ATP production but also decrease their stress resistance. As the respiration rate decreases, so do the levels of enzymes catalyzing rate-determining reactions of the tricarboxylic-acid cycle (providing NADH for respiration) and of mitochondrial folate-mediated NADPH production (required for oxidative defense). The findings demonstrate that exponential growth can represent not a single metabolic/physiological state but a continuum of changing states and that aerobic glycolysis can reduce the energy demands associated with respiratory metabolism and stress survival.
8. El bioreactor LAMBDA MINIFOR es usado para crecer bacterias orales (Streptococcus oralis, Actinomyces naeslundii, Veillonella parvula, Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis) bajo condiciones de plakton: 2014.
Characterization and application of a flow system for in vitro multispecies oral biofilm formation. Blanc V, Isabal S, Sánchez MC, Llama-Palacios A, Herrera D, Sanz M, León R. J Periodont Res 2014; Journal of Periodontal Research. Keywords: modelo de biofilme; clorhexidina; microscopía confocal laser de mapeo scanning; bacterias orales.
Resumen: Bacteria in the oral cavity grow in the form of biofilms; these structures are subject to constant saliva or gingival crevicular fluid flow conditions. The aims of this study were: (i) to develop and to characterize an in-vitro biofilm model with oral bacteria growing under flow and shear conditions; and (ii) to demonstrate the usefulness of the model for evaluating the activity of three antiplaque agents.We used a bioreactor to grow the oral bacteria Streptococcus oralis, Actinomyces naeslundii, Veillonella parvula, Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis under planktonic conditions. Biofilms were established using a modified Robbins device on hydroxyapatite (HAP) discs. [showhide type="link" more_text="Leer más" less_text="Ocultar"]Three- to 7-d-old biofilms were analysed using culture methods, scanning electron microscopy, Live/Dead staining and fluorescence in-situ hybridization (confocal laser scanning microscopy). Finally, we assessed the antimicrobial activity of three mouthrinses [0.12% chlorhexidine (CHX), 0.12% chlorhexidine and sodium fluoride (CHX+NaF) and 0.12% chlorhexidine and 0.05% cetylpyridinium chloride (CHX+CPC)] using a planktonic test (short interval–killing test) and in our 4-d biofilm model. The viable cell counts showed that each species was consistently found in the biofilms throughout the study. The architecture and cell distribution were similar to those described for biofilms in situ, with the exception of a thin layer of living cells that was found close to the HAP. The effectiveness test of the mouthwashes demonstrated that cells in biofilms showed more tolerance compared with planktonic cells. Moreover, it was observed that in 4-d biofilm formed in vitro, CHX+CPC caused significantly higher mortality compared with CHX (p = 0.003) and CHX+NaF (p < 0.001). Our results suggest that we have a highly reproducible system for multispecies oral biofilm formation and that it is a useful tool for assessing antibacterial molecules before their clinical evaluation. It also has great potential to be used in basic research on supragingival and subgingival biofilms.
9. Cultivo de la microalga (Chlorella vulgaris Beyerinck) en el bioreactor de laboratorio MINIFOR: 2014.
Mikrovetikate Chlorella vulgaris kasvatamise eelkatsed bioreaktoris Lambda Minifor
Heitur, Heiko. "Mikrovetika Chlorella vulgaris Beyerincki kasvatamine CO2 sidumise eesmärgil." PhD diss., 2014. Estonian University of Life Sciences, Estonia.
10. Fermentación en lote, selectiva y no-selectiva de extractos de dátiles usando Saccharomyces cerevisiae (línea comercial usada en reposterías (línea silvestre), líneas glucosa selectivas ATCC 36858 y ATCC 36859) en el fermentador LAMBDA MINIFOR: 2014.
Selective fermentation of pitted dates by S. cerevisiae for the production of concentrated fructose syrups and ethanol, Meilana Dharma Putra, Ahmed E. Abasaeed, Mohamed A. Zeinelabdeen, Mohamed H. Gaily, Ashraf K. Sulieman, Journal of Physics: Conference Series 495 (2014) 012034 Chemical Engineering Department, King Saud University, Saudi Arabia. Palabras claves: Fermentación selectiva y no selectiva, levadura, S. cerevisiae, fructosa, etanol, dátiles, HPLC, perfil cinético, lote.
Resumen: About half of worldwide production of dates is unconsumed. Dates contain over 75 % reduced sugars (mostly glucose and fructose with nearly equal amount). Compared to the commercial Saccharomyces cerevisiae wild strain, the strains ATCC 36858 and 36859 could produce high concentration fructose syrups. The fructose fractions obtained were 95.9 and 97.4% for ATCC 36858 and 86.5 and 91.4% for ATCC 36859 at 30 and 33°C, respectively. Fructose yields higher than 90% were obtained using ATCC 36858 compared to those obtained using ATCC 36859 which were 87.3 and 66.1% at 30 and 33°C, respectively. The ethanol yield using ATCC 36858 was higher than that using ATCC 36859 by 16 and 9% at 30 and 33°C, respectively. Through this finding, the production of fructose and ethanol from date extract is a promising process. Moreover, the fructose fractions obtained here (about 90%) are much higher than those obtained with the commercial process, i.e. 55 % fructose syrups.
11. Crecimiento controlado de Staphylococcus aureus en varias concentraciones de BAC (cloruro de benzalconio) en el fermentador MINIFOR. 2013.
The Role of the qacA Gene in Mediating Resistance to Quaternary Ammonium Compounds. Dana Cervinkova, Vladimir Babak, Durdica Marosevic, Iva Kubikova, and Zoran Jaglic. Microbial Drug Resistance. June 2013, 19(3): 160-167. Veterinary Research Institute, Brno, Czech Republic. Palabras claves: Staphylococcus aureus, cloruro de benzalconio (BAC), fase exponencial, expresión, PCR en tiempo real, cultivo, concentración
Resumen: Conditions facilitating resistance to quaternary ammonium compounds (QACs) were investigated in Staphylococcus aureus SK982 exposed to benzalkonium chloride (BAC; a member of QACs) under various circumstances. S. aureus SK982 carrying the qacA gene encoding for resistance to QACs was grown in the presence of stable or gradually increasing concentrations of BAC, or it was exposed to this antiseptic in the exponential phase of growth. Bacteria cultivated in the highest BAC concentrations that did not retard their growth comparing to the untreated control were subjected to real-time quantitative polymerase chain reaction analysis for relative expression of the efflux genes qacA and norA. Under such conditions, S. aureus SK982 tolerated a relatively low stable concentration of BAC (1.22 mg/L) when compared with a gradually increasing antiseptic concentration (tolerance of 4.88 mg/L). However, in both cases, qacA expression was not significant. [showhide type="link" more_text="Leer más" less_text="Ocultar"]The culture exposed in the exponential phase of growth tolerated the highest concentration of BAC (9.76 mg/L) as also accompanied by significant overexpression of qacA. Expression of norA was relatively low regardless of the conditions tested. It seems that under the short-term conditions, the phase of bacterial growth is more important for the expression of BAC resistance than the capability to adapt to this antiseptic. This study provides a deeper insight into the relevance of the qac genes in conferring resistance to QACs.
12. Sistemas para el crecimiento de hibridomas en alta densidad y cultivos celular para la producción de anticuerpos monoclonales (mAbs) con alto rendimiento: Bioreactores de tanque agitado a escala de mesa, 1-5 L (MINIFOR - LAMBDA Laboratory Instruments). 2013.
Gary C. Howard, Matthew R. Kaser. 2013. Making and using antibodies - a practical handbook. CRC Press. Boca Raton. ISBN: 1439869081, 9781439869086.
CRC Press, 2013.
Resumen: Antibodies protect us from a wide range of infectious diseases and cancers and have become an indispensable tool in science—both for conventional immune response research as well as other areas related to protein identification analysis. This second edition of Making and Using Antibodies: A Practical Handbook provides clear guidance on all aspects of how to make and use antibodies for research along with their commercial and industrial applications... From the eradication of smallpox to combating cancer, antibodies present an attractive solution to a range of biomedical problems. They are relatively easy to make and use, have great flexibility in applications, and are cost effective for most labs. This volume will assist biomedical researchers and students and pave the way for future discovery of new methods for making and using antibodies for a host of applications.
13. La respuesta de estrés metabólico en cultivo de células de tomate a baja concentración de oxígeno usando el bioreactor LAMBDA MINIFOR. 2013.
The metabolic response of cultured tomato cells to low oxygen stress. Ampofo-Asiama, J., Baiye, V. M. M., Hertog, M. L. A. T. M., Waelkens, E., Geeraerd, A. H., Nicolai, B. M. (2013), Plant Biology. German Botanical Society and The Royal Botanical Society of the Netherlands. Palabras claves: 3C marcaje; cultivo celular, estrés a baja concentración de O2; Lycopersicum esculentum; metaboloma
Resumen: The storage of fruits and vegetables under a controlled atmosphere can induce low oxygen stress, which can lead to post-harvest losses through the induction of disorders such as core breakdown and browning. To gain better understanding of the metabolic response of plant organs to low oxygen, cultured tomato cells (Lycopersicum esculentum) were used as a model system to study the metabolic stress response to low oxygen (0 and 1 kPa O2). By adding 13C labelled glucose, changes in the levels of polar metabolites and their 13C label accumulation were quantified. Low oxygen stress altered the metabolite profile of tomato cells, with the accumulation of the intermediates of glycolysis in addition to increases in lactate and sugar alcohols. 13C label data showed reduced label accumulation in almost all metabolites except lactate and some sugar alcohols. The results showed that low oxygen stress in tomato cell culture activated fermentative metabolism and sugar alcohol synthesis while inhibiting the activity of the TCA cycle and the biosynthesis of metabolites whose precursors are derived from central metabolism, including fluxes to most organic acids, amino acids and sugars.
14. LAMBDA MINIFOR Bioreactor utilizado para la expresión de proteína recombinante (Chemokines) en E. coli. 2013.
PhD Thesis. 2013. Birgit Kramp, "Establishing the interaction between the CC chemokine ligand 5 and the receptors CCR1 and CCR5
Fakultät für Mathematik, Informatik und Naturwissenschaften, RWTH Aachen, Belgium.
15. Expresión recombinante de Met-CCL5, proteasa resistente CXCL12 (S4V) y F1-CX3CL1 en E. coli para estudiar su rol en la enfermedad cardiovascular (CVD). 2013.
PhD Thesis. 2013. Projahn, D., Generation, function and therapeutic application of chemotactic cytokines in cardiovascular diseases
Institut für Molekulare Herz-Kreislaufforschung, Fakultät für Mathematik, Informatik und Naturwissenschaften, RWTH Aachen, Belgium. Fakultät für Mathematik, Informatik und Naturwissenschaften, RWTH Aachen, Belgium.
16. Expresión de la proteína Caf1 usando la línea Escherichia coli para estudiar la adhesión de células de mamíferos, forma y números de focos de adhesión. 2013.
PhD Thesis. 2013. Machado Roque, A. I., Protein scaffolds for cell culture, 2013 , Newcastle University
17. MINIFOR Bioreactores para el cultivo de células madres. 2012.
A non-rotational, computer-controlled suspension bioreactor for expansion of umbilical cord blood mononuclear cells. Shayan, N., Ebrahimi, M., Beiki, B. and Janzamin, E. 2012. BIOTECHNOLOGY LETTERS. Department of Regenerative Medicine and Stem Cells and Developmental Biology, Cell Research Center, Royan Institute for Stem Cell Biology and Technology, Royan Cord Blood Bank, Tehran, Iran. Palabras claves: Cultivo estático; suspensión celular, sangre del cordón unbical y mezclado vertical
Resumen: The proliferation and differentiation characteristics of umbilical cord blood mononuclear cells were examined in a non-rotational suspension bioreactor with a fishtail mixer. The system consisted of a glass vessel, a mixer that moved vertically, a data acquisition and control system to continuously monitor pH, temperature and dissolved O2. The bioreactor provided superior expansion of total HSCs and not total cell number, as well as expression of stemness-related genes which followed with increasing in number of colony-forming cells during 14 days of culture compared to T -lask culture. Vertical agitation thus reduces the total cell number, which may be related to increased shear stress, but has no effect on HSC function.
18. Producción efectiva de biobutanol a partir de desechos agrícolas. 2012.
Biobutanol Production from Agricultural Waste: A Simple Approach for Pre-Treatment and Hydrolysis. Mežule, L., Tihomirova, K., Neščerecka, A., Juhna, T. Latvian Journal of Chemistry, 2012, 51, No. 4. pp.407-414. Riga Technical University, Latvia Palabras claves: biofuel, biobutanol, desechos agrícolas, hidrósis
Resumen: One of the main concerns regarding extensive production of bio-butanol has been associated with the high costs of the substrate (preparation of fermentable sugars) and the relatively low tolerance of Clostridia to butanol. In this study a simple, mild approach was tested to obtain fermentable sugars from agricultural waste. Giant hogweed and hay was pre-treated with simple boiling and enzymatically hydrolysed. The results demonstrated that after adaptation of the genus Clostridium bacteria to the new substrate, the growth kinetics and sugar consumption of these bacteria we are similar to the ones obtained in traditional culture media.
19. Producción de bioetanol usando levaduras (S. cerevisiae) en el fermentador LAMBDA MINIFOR. 2011.
Effect of wheat gluten proteins on bioethanol yield from grain. Burešová, I., Hřivna, L., Applied Energy Volume 88, Issue 4, April 2011, Pages 1205–1210 Agrotest Fyto, Ltd., Kroměříž, Czech Republic; Mendel University in Brno, Brno, Czech Republic Palabras claves: Bioetanol; Triticale; trigo; Gluten; Proteéna
Resumen: Bioethanol can be used as motor fuel and/or as a gasoline enhancer. A high yield feedstock for bioethanol production is cereal grain. Cereal grains containing less gluten proteins (glutenin and gliadin), but high starch, are favoured by distillers because they increase the bioethanol conversion. The direct effect of wheat gluten proteins on bioethanol yield was studied on triticale grain. Examined triticale Presto 1R.1D5+10-2 and Presto Valdy were developed by introducing selected segments of wheat chromosome 1D into triticale chromosome 1R. Even if the samples analysed in this study do not afford to make definitive assumptions, it can be noticed that in analysed cases the presence of gliadin had more significant effect on investigated parameters than the presence of glutenin. Despite the presence of glutenin subunits did not significantly decrease the investigated parameters – specific weight, [showhide type="link" more_text="Leer más" less_text="Ocultar"]Hagberg falling number and starch content in grain met the requirements for grain for bioethanol production – protein content was higher than is optimal. The fermentation experiments demonstrated good bioethanol yields but depression in grain yields caused by the presence of wheat gliadin and glutenin decreased the energy balance of Presto Valdy and Presto 1R.1D5+10-2.
20. Fermentación anaeróbica del componente de glucosa en los extractos de dátiles por la levadura Saccharomyces cerevisiae. 2010.
A Direct Process for the Production of High Fructose Syrups from Dates Extracts; International Journal of Food Engineering. Mohamed H. Gaily, Basheir M. Elhassan, Ahmed E Abasaeed, Saeed M. Al-Zahrani. Volume 6, Issue 3, ISSN (Online) 1556-3758 King Saud University, Saudi Arabia; University of Khartoum, Sudan Palabras claves: dátiles, fructosa, glucosa, etanol, fermentación, S. Cerevisiae, levadura, mesófilo, lote
Resumen: A process comprising of extraction, fermentation, separation and decolorization for the production of high fructose syrups and/or ethanol from dates' extract was developed. In this paper, selective fermentation of the glucose component in dates' extract by Saccharomyces cerevisiae was employed to tailor the composition of the final product (fructose- or ethanol-rich). Over 90% of dates sugar was extracted by water. The fermentation experiments were conducted at 30 °C and 120 rpm. The commonly marketed 55% fructose syrup was obtained at two operating conditions, i.e., low (<50%) and high (>92%) sugar conversions. Maximum fructose concentrations (77.2-78.2%) were obtained at moderately high sugar conversions (74-82%). Decolorization by activated carbon for the fructose-rich syrup resulted in 87% of color removal. Preliminary economic analysis revealed the viability of this process.
21. Expresión anaeróbica usando el LAMBDA MINIFOR. 2007.
Glyceraldehyde-3-Phosphate Ferredoxin Oxidoreductase from Methanococcus maripaludis. Park, M. -O., Mizutani, T. and Jones, P. R. 2007. Journal of Bacteriology, Tokyo, Japan. Research and Development Division, Fujirebio, Inc., 51 Komiya-cho, Hachioji-shi, Tokyo 192-0031, Japan.
Resumen: The genome sequence of the non-sugar-assimilating mesophile Methanococcus maripaludis contains three genes encoding enzymes: a nonphosphorylating NADP+-dependent glyceraldehyde-3-phosphate dehydrogenase (GAPN), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and glyceraldehyde-3-phosphate ferredoxin oxidoreductase (GAPOR); all these enzymes are potentially capable of catalyzing glyceraldehyde-3-phosphate (G3P) metabolism. GAPOR, whose homologs have been found mainly in archaea, catalyzes the reduction of ferredoxin coupled with oxidation of G3P. GAPOR has previously been isolated and characterized only from a sugar-assimilating hyperthermophile, Pyrococcus furiosus (GAPORPf), and contains the rare metal tungsten as an irreplaceable cofactor. Active recombinant M. maripaludis GAPOR (GAPORMm) was purified from Escherichia coli grown in minimal medium containing 100 μM sodium molybdate. [showhide type="link" more_text="Leer más" less_text="Ocultar"]In contrast, GAPORMm obtained from cells grown in medium containing tungsten (W) and W and molybdenum (Mo) or in medium without added W and Mo did not display any activity. Activity and transcript analysis of putative G3P-metabolizing enzymes and corresponding genes were performed with M. maripaludis cultured under autotrophic conditions in chemically defined medium. The activity of GAPORMm was constitutive throughout the culture period and exceeded that of GAPDH at all time points. As GAPDH activity was detected in only the gluconeogenic direction and GAPN activity was completely absent, only GAPORMm catalyzes oxidation of G3P in M. maripaludis. Recombinant GAPORMm is posttranscriptionally regulated as it exhibits pronounced and irreversible substrate inhibition and is completely inhibited by 1 μM ATP. With support from flux balance analysis, it is concluded that the major physiological role of GAPORMm in M. maripaludis most likely involves only nonoptimal growth conditions.
22. pH y temperatura fueron registrados de forma continua con el LAMBDA MINIFOR y el SIAM software. 2005.
Photochemical Reactivity of Trifluoromethyl Aromatic Amines: The Example of 3,5-diamino-trifluoromethyl-benzene (3,5-DABTF) Chaignon, P., Cortial, S., Guerineau, V., Adeline, M. T., Giannotti, C., Fan, G. and Ouazzani, J. 2005. Photochem Photobiol Volume 81, Issue 6, pages 1539–1543, November 2005. Institut de Chimie des Substances Naturelles, C.N.R.S, Avenue de la Terrasse, 91198, Gif-sur-Yvette Cedex. France.
Resumen: This work presents the application of an on-line photoreactor to the detection of 3,5-diamino-trifluoromethyl-benzene (3,sDABTF) in aqueous solutions. When irradiated at 310 nm, this compound is defluorinated to 3,s-diaminobenzoic acid by a nucleophilic substitution of the fluoride by water. Concomitantly, defluorination intermediates polymerize through amide bonds to give dark-colored compounds. We take advantage of the photocatalyzed defluorination and the subsequent decrease in pH to develop an original and specific photoreactor. Continuous recording of pH and temperature in the outlet of the reactor by a dual electrode gives us an opportunity to optimize the system. In the photoreactor, 3,5-DABTF is immediately and totally transformed as attested by the rapid drop of the flowing solution pH from 6.2 to 3.2 and the chromatographic analysis of the outgoing solutions. The detection remains effective from 1 to 1000 parts per million.
23. Bioreactores - Un resumen de las innovaciones instrumentadas en el bioreactor MINIFOR. 2003.
Lehky, P. 2003. Bioreactors - New Solutions for Old Problems. International Congress on Bioreactor Technology, Tampere, Finland. Proceedings of the 1st International Congress on Bioreactor Technology in Cell, Tissue Culture and Biomedical Applications: Tampere, Finland, 14-18 July, 2003. ISBN: 9529930208, 9789529930203. Palabras claves: bioreactor, fermentador, cultivo celular, electrodo DO, caudal del flujo de gas, estación de gases.
Resumen: A major effort has been invested in making bioreactors more efficient. The goal has been achieved in several steps. Firstly, a replacement of the traditional rotation movement by an oscillating up and down movement of stirring discs driven by an electromagnet yields several important advantages. A soft, whole volume movement of the medium results in optimal gas distribution throughout the medium and is also advantageous for cell growth. No vortex is formed and the elimination of baffles simplifies the construction of the bioreactor. A single silicone membrane efficiently replaces traditional seals and assures perfect sterility. In the case of extremely sensitive cells gas distribution tubing can be wound on a spiral fixed to the axis. The up and down movement facilitates the gas transfer and simultaneously provides a gentle movement of the medium. Secondly, a new heat radiation system has also been introduced. A heating spiral in a gilded parabolic reflector has been placed under the bottom of the vessel. The IR rays efficiently heat up the culture without overheating it at any volume of medium. Several innovations allow the construction of high quality bioreactors at lower cost.