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1.
Braz J Microbiol ; 55(2): 1167-1177, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38557863

RESUMO

It is essential to evaluate the effects of operating conditions in submerged cultures of filamentous microorganisms. In particular, the impeller type influences the flow pattern, power consumption, and energy dissipation, leading to differences in the hydrodynamic environment that affect the morphology of the microorganism. This work investigated the effect of different impeller types, namely the Rushton turbine (RT-RT) and Elephant Ear impellers in up-pumping (EEUP) and down-pumping (EEDP) modes, on cellular morphology and clavulanic acid (CA) production by Streptomyces clavuligerus in a stirred-tank bioreactor. At 800 rpm and 0.5 vvm, the cultivations performed using RT-RT and EEUP impellers provided higher shear conditions and oxygen transfer rates than those observed with EEDP. These conditions resulted in higher clavulanic acid production using RT-RT (380.7 mg/L) and EEUP (453.3 mg/L) impellers, compared to EEDP (196.6 mg/L). Although the maximum CA concentration exhibited the same order of magnitude for RT-RT and EEUP impellers, the latter presented 40% of the specific power consumption (4.9 kW/m3) compared to the classical RT-RT (12.0 kW/m3). The specific energy for CA production ( E CA ), defined as the energy cost to produce 1 mg of CA, was 3.5 times lower using the EEUP impeller (1.91 kJ/mgCA) when compared to RT-RT (5.91 kJ/mgCA). Besides, the specific energy for O2 transfer ( E O 2 ), the energy required to transfer 1 mmol of O2, was 2.3 times lower comparing the EEUP impeller (3.28 kJ/mmolO2) to RT-RT (7.65 kJ/mmolO2). The results demonstrated the importance of choosing the most suitable impeller configuration in conventional bioreactors to manufacture bioproducts.


Assuntos
Reatores Biológicos , Ácido Clavulânico , Streptomyces , Ácido Clavulânico/biossíntese , Streptomyces/metabolismo , Streptomyces/crescimento & desenvolvimento , Reatores Biológicos/microbiologia , Fermentação , Antibacterianos/biossíntese
2.
Nat Prod Res ; 38(10): 1662-1669, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-37211779

RESUMO

Microbial lipids are a valuable source of potential biofuels and essential polyunsaturated fatty acids. The optimization of the fermentation conditions is a strategy that affects the total lipid concentration. The genus Nigrospora sp. has been the target of investigations based on its potential bioherbicidal action. Therefore, this study developed a strategy to maximize the biomass concentration and lipid accumulation by Nigrospora sp. in submerged fermentation. Different media compositions and process variables were investigated in shaken flasks and bioreactor in batch and fed-batch modes. Maximum biomass concentration and lipid accumulations were 40.17 g/L and 21.32 wt% in the bioreactor, which was 2.1 and 5.4 times higher than the same condition in shaken flasks, respectively. This study presents relevant information to the production of fungal lipids since few investigations are exploring the fed-batch strategy to increase the yield of fungi lipids, as well as few studies investigating Nigrospora sp. to produce lipids.


Assuntos
Ascomicetos , Reatores Biológicos , Fermentação , Lipídeos , Biomassa , Biocombustíveis
3.
Mol Biotechnol ; 66(2): 354-364, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-37162721

RESUMO

This work aimed to assess the Sf9 cell metabolism during growth, and infection steps with recombinant baculovirus bearing rabies virus proteins, to finally obtain rabies VLP in two culture systems: Schott flask (SF) and stirred tank reactor (STR). Eight assays were performed in SF and STR (four assays in each system) using serum-free SF900 III culture medium. Two non-infection growth kinetics assays and six recombinant baculovirus infection assays. The infection runs were carried out at 0.1 pfu/cell multiplicity of infection (MOI) for single baculovirus bearing rabies glycoprotein (BVG) and matrix protein (BVM) and a coinfection with both baculoviruses at MOI of 3 and 2 pfu/cell for BVG and BVM, respectively. The SF assays were done in triplicate. The glucose, glutamine, glutamate, lactate, and ammonium uptake or release specific rates were quantified over the exponential growth phase and infection stage. The highest uptake specific rate was observed for glucose (42.5 × 10-12 mmol cell/h) in SF and for glutamine (30.8 × 10-12 mmol/cell/h) in STR, in the exponential growth phases. A wave pattern was observed for assessed analytes throughout the infection phase and the glucose had the highest wave amplitude within the 10-10 mmol cell/h order. This alternative uptake and release behavior is in harmony with the lytic cycle of baculovirus in insect cells. The virus propagation and VLP generation were not limited by glucose, glutamine, and glutamate, neither by the toxicity of lactate nor ammonium under the conditions appraised in this work. The findings from this work can be useful to set baculovirus infection processes at high cell density to improve rabies VLP yield, purity, and productivity.


Assuntos
Compostos de Amônio , Vírus da Raiva , Raiva , Animais , Células Sf9 , Vírus da Raiva/genética , Glutamina , Baculoviridae/genética , Proteínas Recombinantes/genética , Meios de Cultura Livres de Soro , Ácido Glutâmico , Lactatos , Glucose , Spodoptera
4.
Front Bioeng Biotechnol ; 11: 1324417, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-38152287

RESUMO

The presence of sulfur impurities in complex iron ores represents a significant challenge for the iron mining and steel-making industries as their removal often necessitates the use of hazardous chemicals and energy-intensive processes. Here, we examined the microbial and mineralogical composition of both primary and secondary iron concentrates, identifying the presence of Sulfobacillus spp. and Leptospirillum spp., while sulfur-oxidizing bacteria were absent. We also observed that these concentrates displayed up to 85% exposed pyrrhotite. These observations led us to explore the capacity of Acidithiobacillus thiooxidans to remove pyrrhotite-sulfur impurities from iron concentrates. Employing stirred tank bioreactors operating at 30°C and inoculated with 5·106 (At. thiooxidans cells mL-1), we achieved 45.6% sulfur removal over 16 days. Then, we evaluated packed leaching columns operated at 30°C, where the At. thiooxidans enriched system reached 43.5% desulfurization over 60 days. Remarkably, sulfur removal increased to 80% within 21 days under potassium limitation. We then compared the At. thiooxidans-mediated desulfurization process, with and without air supply, under potassium limitation, varying the initial biomass concentration in 1-m columns. Aerated systems facilitated approximately 70% sulfur removal across the entire column with minimal iron loss. In contrast, non-aerated leaching columns achieved desulfurization levels of only 6% and 26% in the lower and middle sections of the column, respectively. Collectively, we have developed an efficient, scalable biological sulfur-removal technology for processing complex iron ores, aligning with the burgeoning demand for sustainable practices in the mining industry.

5.
Mol Biotechnol, v. 66, p. 354-364, abr. 2023
Artigo em Inglês | Sec. Est. Saúde SP, SESSP-IBPROD, Sec. Est. Saúde SP | ID: bud-4915

RESUMO

This work aimed to assess the Sf9 cell metabolism during growth, and infection steps with recombinant baculovirus bearing rabies virus proteins, to fnally obtain rabies VLP in two culture systems: Schott fask (SF) and stirred tank reactor (STR). Eight assays were performed in SF and STR (four assays in each system) using serum-free SF900 III culture medium. Two non-infection growth kinetics assays and six recombinant baculovirus infection assays. The infection runs were carried out at 0.1 pfu/cell multiplicity of infection (MOI) for single baculovirus bearing rabies glycoprotein (BVG) and matrix protein (BVM) and a coinfection with both baculoviruses at MOI of 3 and 2 pfu/cell for BVG and BVM, respectively. The SF assays were done in triplicate. The glucose, glutamine, glutamate, lactate, and ammonium uptake or release specifc rates were quantifed over the exponential growth phase and infection stage. The highest uptake specifc rate was observed for glucose (42.5× 10–12 mmol cell/h) in SF and for glutamine (30.8× 10–12 mmol/cell/h) in STR, in the exponential growth phases. A wave pattern was observed for assessed analytes throughout the infection phase and the glucose had the highest wave amplitude within the 10–10 mmol cell/h order. This alternative uptake and release behavior is in harmony with the lytic cycle of baculovirus in insect cells. The virus propagation and VLP generation were not limited by glucose, glutamine, and glutamate, neither by the toxicity of lactate nor ammonium under the conditions appraised in this work. The fndings from this work can be useful to set baculovirus infection processes at high cell density to improve rabies VLP yield, purity, and productivity.

6.
Vaccines, v. 11, n. 9, 39, 2023.
Artigo em Inglês | Sec. Est. Saúde SP, SESSP-IBPROD, Sec. Est. Saúde SP | ID: bud-4762

RESUMO

This work aimed to assess, following upstream optimization in Schott flasks, the scalability from this culture platform to a stirred-tank bioreactor in order to yield rabies-recombinant baculovirus, bearing genes of G (BVG) and M (BVM) proteins, and to obtain rabies virus-like particles (VLP) from them, using Sf9 insect cells as a host. Equivalent assays in Schott flasks and a bioreactor were performed to compare both systems and a multivariate statistical approach was also carried out to maximize VLP production as a function of BVG and BVM’s multiplicity of infection (MOI) and harvest time (HT). Viable cell density, cell viability, virus titer, BVG and BVM quantification by dot-blot, and BVG quantification by Enzyme-Linked Immunosorbent Assay (ELISA) were monitored throughout the assays. Furthermore, transmission electron microscopy was used to characterize rabies VLP. The optimal combination for maximum VLP expression was BVG and BVM MOI of 2.3 pfu/cell and 5.1 pfu/cell, respectively, and 108 h of harvest time. The current study confirmed that the utilization of Schott flasks and a benchtop bioreactor under the conditions applied herein are equivalent regarding the cell death kinetics corresponding to the recombinant baculovirus infection process in Sf9 cells. According to the results, the hydrodynamic and chemical differences in both systems seem to greatly affect the virus and VLP integrity after release.

7.
Proc Bioch, v. 124, p. 189-200, jan. 2023
Artigo em Inglês | Sec. Est. Saúde SP, SESSP-IBPROD, Sec. Est. Saúde SP | ID: bud-4718

RESUMO

The technologies used in rabies vaccines manufacturing for human use are based on the inactivated virus platform. An alternative to traditional vaccines is virus-like particles (VLPs). This work aimed to characterize the oxygen uptake and transfer rate parameters throughout recombinant baculovirus (rBV) and rabies VLPs production using Sf9 cells in stirred tank bioreactor (STB) for a better bioprocess understanding and scalability. Four runs in a bench STB were performed: cell culture without infection; cells infected singly with rBV bearing rabies virus glycoprotein (rBVG, multiplicity of infection, MOI=0.1 pfu/cell) and matrix protein (rBVM, MOI=0.1 pfu/cell), and coinfected with BVG and BVM at MOI of 3 and 2 pfu/cell, respectively. The specific oxygen uptake rate () and volumetric oxygen transfer coefficient () were monitored throughout the reactions, as well as viable cell concentration, viability, rBV titers, and protein concentration. According to the results herein, the aeration and agitation systems in a bioreactor at a higher scale could be designed using the criterium for scale-up of constant , without oxygen facilities. Besides, rabies VLPs volumetric yield of 2.8 mg/L with a typical size (55–68 nm) was obtained. These findings suggest a promising bioprocess for rabies VLPs at a commercial scale.

8.
Bioprocess Biosyst Eng ; 45(10): 1635-1644, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-35974197

RESUMO

L-Asparaginase (L-ASNase) is an enzyme applied in the treatment of lymphoid malignancies. However, an innovative L-ASNase with high yield and lower side effects than the commercially available preparations are still a market requirement. Here, a new-engineered Bacillus subtilis strain was evaluated for Aliivibrio fischeri L-ASNase II production, being the bioprocess development and the enzyme characterization studied. The pBS0E plasmid replicative in Bacillus sp and containing PxylA promoter inducible by xylose and its repressive molecule sequence (XylR) was used for the genetic modification. Initially, cultivations were carried out in orbital shaker, and then the process was scaled up to stirred tank bioreactor (STB). After the bioprocess, the cells were recovered and submitted to ultrasound sonication for cells disruption and intracellular enzyme recovery. The enzymatic extract was characterized to assess its biochemical, kinetic and thermal properties using L-Asparagine and L-Glutamine as substrates. The results indicated the potential enzyme production in STB achieving L-ASNase activity up to 1.539 U mL-1. The enzymatic extract showed an optimum pH of 7.5, high L-Asparagine affinity (Km = 1.2275 mmol L-1) and low L-Glutaminase activity (0.568-0.738 U mL-1). In addition, thermal inactivation was analyzed by two different Kinect models to elucidate inactivation mechanisms, low kinetic thermal inactivation constants for 25 ºC and 37 ºC (0.128 and 0.148 h-1, respectively) indicate an elevated stability. The findings herein show that the produced recombinant L-ASNase has potential to be applied for pharmaceutical purposes.


Assuntos
Antineoplásicos , Produtos Biológicos , Aliivibrio fischeri , Antineoplásicos/química , Asparaginase/química , Asparaginase/genética , Asparaginase/uso terapêutico , Asparagina , Bacillus subtilis/genética , Glutaminase , Glutamina , Preparações Farmacêuticas , Xilose
9.
J Biosci Bioeng ; 134(1): 21-28, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-35461767

RESUMO

Bacillus circulans E9 (now known as Niallia circulans) promotes plant growth-producing indole-3-acetic acid (IAA), showing potential for use as a biofertilizer. In this work, the use of a low-cost medium containing industrial substrates, soybean, pea flour, Solulys, Pharmamedia, yeast extract, and sodium chloride (NaCl), was evaluated as a substitute for microbiological Luria Broth (LB) medium for the growth of B. circulans E9 and the production of IAA. In Erlenmeyer flasks with pea fluor medium (PYM), the maximum production of IAA was 7.81 ± 0.16 µg mL-1, while in microbiological LB medium, it was 3.73 ± 0.15 µg mL-1. In addition, an oxygen transfer rate (OTR) of 1.04 kg O2 m-3 d-1 allowed the highest bacterial growth (19.3 ± 2.18 × 1010 CFU mL-1) and IAA production (10.7 µg mL-1). Consequently, the OTR value from the flask experiments was used to define the conditions for the operation of a 1 L stirred tank bioreactor. The growth and IAA production of B. circulans cultured in a bioreactor with PYM medium were higher (8 and 1.6 times, respectively) than those of bacteria cultured in Erlenmeyer flasks. IAA produced in a bioreactor by B. circulans was shown to induce the root system in Arabidopsis thaliana, similar to synthetic IAA. The results of this study demonstrate that PYM medium may be able to be used for the mass production of B. circulans E9 in bioreactors, increasing both bacterial growth and IAA production. This low-cost medium has the potential to be employed to grow other IAA-producing bacterial species.


Assuntos
Arabidopsis , Bacillus , Reatores Biológicos , Meios de Cultura , Ácidos Indolacéticos , Cloreto de Sódio
10.
Biopolymers ; 113(6): e23488, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35338709

RESUMO

Produced water (PW) and crude glycerin (CG) are compounds overproduced by the oil and biodiesel industry and significant scientific efforts are being applied for properly recycling them. The aim of this research is to combine such industrial byproducts for sustaining the production of xanthan by Xanthomonas campestris. Xanthan yields and viscosity on distinct PW ratios (0, 10, 15, 25, 50, 100) and on 100% dialyzed PW (DPW) in shaker batch testing identified DPW treatment as the best approach for further bioreactor experiments. Such experiments showed a xanthan yield of 17.3 g/L within 54 h and a viscosity of 512 mPa s. Physical-chemical characterization (energy dispersive X-ray spectroscopy, scanning electron microscopy and Raman spectroscopy) showed similarities between the produced gum and the experimental control. This research shows a clear alternative for upcycling high salinity PW and CG for the generation of a valued bioproduct for the oil industry.


Assuntos
Polissacarídeos Bacterianos , Xanthomonas campestris , Glicerol , Polissacarídeos Bacterianos/química , Viscosidade , Água
11.
Vaccines (Basel) ; 11(1)2022 Dec 24.
Artigo em Inglês | MEDLINE | ID: mdl-36679884

RESUMO

This work aimed to assess, following upstream optimization in Schott flasks, the scalability from this culture platform to a stirred-tank bioreactor in order to yield rabies-recombinant baculovirus, bearing genes of G (BVG) and M (BVM) proteins, and to obtain rabies virus-like particles (VLP) from them, using Sf9 insect cells as a host. Equivalent assays in Schott flasks and a bioreactor were performed to compare both systems and a multivariate statistical approach was also carried out to maximize VLP production as a function of BVG and BVM's multiplicity of infection (MOI) and harvest time (HT). Viable cell density, cell viability, virus titer, BVG and BVM quantification by dot-blot, and BVG quantification by Enzyme-Linked Immunosorbent Assay (ELISA) were monitored throughout the assays. Furthermore, transmission electron microscopy was used to characterize rabies VLP. The optimal combination for maximum VLP expression was BVG and BVM MOI of 2.3 pfu/cell and 5.1 pfu/cell, respectively, and 108 h of harvest time. The current study confirmed that the utilization of Schott flasks and a benchtop bioreactor under the conditions applied herein are equivalent regarding the cell death kinetics corresponding to the recombinant baculovirus infection process in Sf9 cells. According to the results, the hydrodynamic and chemical differences in both systems seem to greatly affect the virus and VLP integrity after release.

12.
Bioprocess Biosyst Eng ; 44(8): 1721-1732, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-33821325

RESUMO

The production of biocompounds through the cultivation of filamentous microorganisms is mainly affected by Oxygen Transfer Rate (OTR) and shear rate ([Formula: see text]) conditions. Despite efforts have been made to evaluate the effect of operating variables (impeller speed, N; and airflow rate, ϕair) on clavulanic acid production, no analysis regarding the effect of OTR and [Formula: see text] was made. Then, the aim of this study was to evaluate the dissociated effect of physical phenomena such as oxygen transfer and shear rate in the production of clavulanic acid from Streptomyces clavuligerus using a stirred tank bioreactor. Streptomyces clavuligerus cultivations were performed at five different OTR and [Formula: see text] conditions by manipulating the operating conditions (N, ϕair, and gas inlet composition). Cultivations performed at equal impeller speed (600 rpm, similar [Formula: see text]) using oxygen enrichment, showed that CA productivity (ProdCA) was positively affected by OTR increase. Subsequently, the different shear conditions (achieved by varying the impeller speed) lead to an increase in CA production levels. Despite both OTR and shear rate positively enhanced CA productivity, [Formula: see text] exhibited the highest impact: an increase of 145% in OTRinitial enhanced the clavulanic acid productivity of about 29%, while an increment in the shear rate of 134% raised the ProdCA in 53%.


Assuntos
Ácido Clavulânico/química , Microbiologia Industrial/métodos , Oxigênio/química , Streptomyces/metabolismo , Reatores Biológicos , Biotecnologia/métodos , Meios de Cultura , Desenho de Equipamento , Resistência ao Cisalhamento , Fatores de Tempo
13.
J Fungi (Basel) ; 6(4)2020 Nov 02.
Artigo em Inglês | MEDLINE | ID: mdl-33147713

RESUMO

Natural colorants from microbial fermentation have gained significant attention in the market to replace the synthetic ones. Talaromyces spp. produce yellow-orange-red colorants, appearing as a potential microorganism to be used for this purpose. In this work, the production of natural colorants by T. amestolkiae in a stirred-tank bioreactor is studied, followed by its application as additives in bio-based films. The effect of the pH-shift control strategy from 4.5 to 8.0 after 96 h of cultivation is evaluated at 500 rpm, resulting in an improvement of natural colorant production, with this increase being more significant for the orange and red ones, both close to 4-fold. Next, the fermented broth containing the colorants is applied to the preparation of cassava starch-based films in order to incorporate functional activity in biodegradable films for food packaging. The presence of fermented broth did not affect the water activity and total solids of biodegradable films as compared with the standard one. In the end, the films are used to pack butter samples (for 45 days) showing excellent results regarding antioxidant activity. It is demonstrated that the presence of natural colorants is obtained by a biotechnology process, which can provide protection against oxidative action, as well as be a functional food additive in food packing biomaterials.

14.
Braz. arch. biol. technol ; Braz. arch. biol. technol;63: e20190015, 2020. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1132271

RESUMO

Abstract (1) Background: Oxygen supply is an important parameter to be considered in submerged cultures. This study evaluated the influence of different conditions for dissolved oxygen (DO) concentration on laccases activities and growth of Pleurotus sajor-caju PS-2001 in submerged process in stirred-tank bioreactor. (2) Methods: Initially, three different conditions were tested: uncontrolled DO and minimum levels of 30% and 80% of saturation, with the pH controlled between 4.5 and 7.0. (3) Results: Best results were observed at 30% DO (26 U mL-1 of laccases at 96 h), whereas higher mycelial biomass was observed at 30% and 80% DO (above 4.5 g L-1). Four different conditions of DO (uncontrolled, 10%, 30% and 50% of saturation) were tested at pH 6.5, with higher laccases activity (80 U mL-1 at 66 h) and lower mycelial growth (1.36 g L-1 at 90 h) being achieved with DO of 30%. In this test, the highest values for volumetric productivity and specific yield factor were determined. Under the different pH conditions tested, the production of laccases is favoured at DO concentration of 30% of saturation, while superior DO levels favours fungal growth. (4) Conclusion: The results indicate that dissolved oxygen concentration is a critical factor for the culture of P. sajor-caju PS-2001 and has important effects not only on laccases production but also on fungal growth.


Assuntos
Oxigênio Dissolvido , Biomassa , Reatores Biológicos , Pleurotus/crescimento & desenvolvimento , Pleurotus/enzimologia , Lacase/biossíntese
15.
Braz J Chem Eng, v. 35, n. 2, p. 441-458
Artigo em Inglês | Sec. Est. Saúde SP, SESSP-IBPROD, Sec. Est. Saúde SP | ID: bud-2580

RESUMO

Baby Hamster Kidney cells (BHK-21) are commonly used in research and the biopharmaceutical industry. This work aimed to model the kinetic performance in batch operation mode of BHK-21 cells cultured in two stirred tank configurations using different dissolved oxygen concentrations and pH control strategies. Viable and dead cell concentrations, as well as glucose, glutamine, lactate and ammonium concentrations, were monitored. Statistical multiple linear regression, logistic equation and multiplicative Monod kinetic models were fitted. Statistical models for viable cells concentration as a function of nutrient and metabolite concentrations were significant (R2 >0.91). Logistic model parameters: intrinsic growth rate, cell density level in the medium and time for reaching maximum cell concentrations were within 0.061-0.083 h-1, 1.85-5.39 x 109 cell L-1 and 52-90 h ranges, respectively. A Monod-type model was satisfactorily fitted to the experimental data. Relative errors were lower than 10% for six monitored state variables in most of the assessed experimental conditions. The three models developed in this work can be used in bioprocesses involving BHK-21 with good fitting.

16.
Biotechnol J ; 11(8): 1048-59, 2016 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-27168373

RESUMO

Mesenchymal stem/stromal cells (MSC) are being widely explored as promising candidates for cell-based therapies. Among the different human MSC origins exploited, umbilical cord represents an attractive and readily available source of MSC that involves a non-invasive collection procedure. In order to achieve relevant cell numbers of human MSC for clinical applications, it is crucial to develop scalable culture systems that allow bioprocess control and monitoring, combined with the use of serum/xenogeneic (xeno)-free culture media. In the present study, we firstly established a spinner flask culture system combining gelatin-based Cultispher(®) S microcarriers and xeno-free culture medium for the expansion of umbilical cord matrix (UCM)-derived MSC. This system enabled the production of 2.4 (±1.1) x10(5) cells/mL (n = 4) after 5 days of culture, corresponding to a 5.3 (±1.6)-fold increase in cell number. The established protocol was then implemented in a stirred-tank bioreactor (800 mL working volume) (n = 3) yielding 115 million cells after 4 days. Upon expansion under stirred conditions, cells retained their differentiation ability and immunomodulatory potential. The development of a scalable microcarrier-based stirred culture system, using xeno-free culture medium that suits the intrinsic features of UCM-derived MSC represents an important step towards a GMP compliant large-scale production platform for these promising cell therapy candidates.


Assuntos
Técnicas de Cultura Celular por Lotes/métodos , Células-Tronco Mesenquimais/citologia , Cordão Umbilical/citologia , Reatores Biológicos , Contagem de Células , Diferenciação Celular , Proliferação de Células , Humanos , Imunofenotipagem , Células-Tronco Mesenquimais/imunologia , Cordão Umbilical/imunologia
17.
Methods Mol Biol ; 1391: 173-86, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27108317

RESUMO

Brugmansia candida (syn. Datura candida) is a South American native plant that produces tropane alkaloids. Hyoscyamine, 6ß-hydroxyhyoscyamine (anisodamine), and scopolamine are the most important ones due to their anticholinergic activity. These bioactive compounds have been historically and widely applied in medicine and their demand is continuous. Their chemical synthesis is costly and complex, and thereby, these alkaloids are industrially produced from natural producer plants. The production of these secondary metabolites by plant in vitro cultures such as hairy roots presents certain advantages over the natural source and chemical synthesis. It is well known that hairy roots produced by Agrobacterium rhizogenes infection are fast-growing cultures, genetically stable and able to grow in hormone-free media. Additionally, recent progress achieved in the scaling up of hairy root cultures makes this technology an attractive tool for industrial processes. This chapter is focused on the methods for the induction and establishment of B. candida hairy roots. In addition, the scaling up of hairy root cultures in bioreactors and tropane alkaloid analysis is discussed.


Assuntos
Biotecnologia/métodos , Datura/fisiologia , Raízes de Plantas/fisiologia , Tropanos/metabolismo , Agrobacterium/crescimento & desenvolvimento , Reatores Biológicos , Biotecnologia/instrumentação , Cromatografia Líquida de Alta Pressão/métodos , Técnicas de Cultura/instrumentação , Técnicas de Cultura/métodos , DNA de Plantas/genética , DNA de Plantas/isolamento & purificação , Datura/genética , Datura/crescimento & desenvolvimento , Datura/microbiologia , Desenho de Equipamento , Melhoramento Vegetal/métodos , Raízes de Plantas/genética , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/microbiologia , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Plantas Geneticamente Modificadas/fisiologia , Esterilização/métodos , Tropanos/análise , Tropanos/isolamento & purificação
18.
Appl Biochem Biotechnol ; 177(6): 1201-15, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26304128

RESUMO

Aspergillus flavipes FP-500 grew up on submerged cultures using lemon peel as the only carbon source, developing several batch and pulsed fed-batch trials on a stirred tank reactor. The effect of carbon source concentration, reducing sugar presence and initial pH on exopectinase and endopectinase production, was analyzed on batch cultures. From this, we observed that the highest substrate concentration favored biomass (X max) but had not influence on the corresponding specific production (q p) of both pectinases; the most acid condition provoked higher endopectinase-specific productions but had not a significant effect on those corresponding to exopectinases; and reducing sugar concentrations higher than 1.5 g/L retarded pectinase production. On the other hand, by employing the pulsed fed-batch operation mode, we observed a prolonged growth phase, and an increase of about twofold on endopectinase production without a significant raise on biomass concentration. So, pulsed fed-batch seems to be a good alternative for obtaining higher endopectinase titers by using high lemon peel quantities without having mixing and repression problems to the system. The usefulness of unstructured kinetic models for explaining, under a theoretic level, the behavior of the fungus along the batch culture with regard to pectinase production was evident.


Assuntos
Aspergillus/crescimento & desenvolvimento , Reatores Biológicos , Citrus/química , Frutas/química
19.
Electron. j. biotechnol ; Electron. j. biotechnol;10(2): 230-239, Apr. 15, 2007. graf, tab
Artigo em Inglês | LILACS | ID: lil-499178

RESUMO

Acremonium chrysogenum NCIM 1069 was used for the biosynthesis of cephalosporin-C (CPC) in batch mode of cultivation. The effect of different medium constituents for better yield of CPC was thoroughly investigated. From the results of the fermentation, it was found that ammonium sulphate as inorganic nitrogen source and methionine at the concentration of 0.4 percent are most suitable for higher yield of antibiotic. The variation in the C/N ratio on the biosynthesis of CPC showed that a C/N ratio of 8.0 is most suitable for maximum production of CPC


Assuntos
Acremonium/metabolismo , Cefalosporinas/biossíntese , Reatores Biológicos , Meios de Cultura , Fermentação , Metionina/metabolismo
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