ABSTRACT
The simulations and predictions obtained from mathematical models of bioprocesses conducted by microorganisms are not overvalued. Mechanistic models are bringing a better process understanding and the possibility of simulating unmeasurable variables. The Dynamic Energy Budget (DEB) model is an energy balance that can be formulated for any living organism and can be classified as a structured model. In this study, the DEB model was used to describe E. coli growth in a batch reactor in carbon and nitrogen substrate limitation conditions. The DEB model provides a possibility to follow the changes in the microbes' cells including their elemental composition and content of some important cell ingredients in different growth phases in substrate limitation conditions which makes it more informative compared to Monod's model. The model can be used as an optimal choice between Monod-like models and flux-based approaches. KEY POINTS: ⢠The DEB model can be used to catch changes in elemental composition of E. coli ⢠Bacteria batch culture growth phases can be explained by the DEB model ⢠The DEB model is more informative compared to Monod's based models.
Subject(s)
Bioreactors , Carbon , Energy Metabolism , Escherichia coli , Nitrogen , Escherichia coli/growth & development , Escherichia coli/metabolism , Nitrogen/metabolism , Carbon/metabolism , Bioreactors/microbiology , Models, Biological , Culture Media/chemistry , Batch Cell Culture Techniques , Models, TheoreticalABSTRACT
The gut microbiota of healthy breastfed infants is often dominated by bifidobacteria. In an effort to mimic the microbiota of breastfed infants, modern formulas are fortified with bioactive and bifidogenic ingredients. These ingredients promote the optimal health and development of infants as well as the development of the infant microbiota. Here, we used INFOGEST and an in vitro batch fermentation model to investigate the gut health-promoting effects of a commercial infant formula supplemented with a blend containing docosahexaenoic acid (DHA) (20 mg/100 kcal), polydextrose and galactooligosaccharides (PDX/GOS) (4 g/L, 1:1 ratio), milk fat globule membrane (MFGM) (5 g/L), lactoferrin (0.6 g/L), and Bifidobacterium animalis subsp. lactis, BB-12 (BB-12) (106 CFU/g). Using fecal inoculates from three healthy infants, we assessed microbiota changes, the bifidogenic effect, and the short-chain fatty acid (SCFA) production of the supplemented test formula and compared those with data obtained from an unsupplemented base formula and from the breast milk control. Our results show that even after INFOGEST digestion of the formula, the supplemented formula can still maintain its bioactivity and modulate infants' microbiota composition, promote faster bifidobacterial growth, and stimulate production of SCFAs. Thus, it may be concluded that the test formula containing a bioactive blend promotes infant gut microbiota and SCFA profile to something similar, but not identical to those of breastfed infants.
Subject(s)
Bifidobacterium animalis , Microbiota , Infant , Female , Humans , Infant Formula , Milk, Human , Dietary Supplements , Breast Feeding , Bifidobacterium , Feces/microbiology , Oligosaccharides/pharmacologyABSTRACT
The rising generation of waste activated sludge (WAS) demands a fundamental shift towards resource reuse and recovery. The conventional methodologies used to manage this by-product derived from wastewater treatment plants are increasingly constrained due to stringent regulatory measures aimed at mitigating its adverse impacts on the environment and public health. Therefore, this work evaluated a promising strategy for the efficient management of WAS, transforming it into a valuable renewable source to produce high-value-added compounds, such as lipids and a slow-release fertilizer (struvite). Wet oxidation (WO) was identified as a suitable technique for solubilising WAS while generating short-chain fatty acids (primarily acetic acid). It was found that conducting WO at 200 °C for 120 min resulted in a 65% reduction of the total suspended solids (TSS) content and 87% of the volatile suspended solids (VSS) content. Additionally, under these conditions, 4440 ± 105 mg/L and 593 ± 21 mg/L of acetic and propionic acid were obtained, respectively, which were assimilated by Yarrowia lipolytica to produce biolipids. Furthermore, the rupture of WAS flocs also led to the solubilisation of 980 ± 8 mg/L of ammonium. During the struvite precipitation stage, a NH4:PO4:Mg ratio of 1:1.5:1.5 was found to be the most effective for removing soluble ammonium (97.4 ± 0.8%), resulting in a high-purity struvite formation, and enhancing the carbon/nitrogen (C/N) ratio of the oxidised WAS from 3 to 105. This improvement in the C/N ratio raised the lipid content from 36 ± 1% to 49 ± 1% during the cultivation of Y. lipolytica. The application of the sequencing batch culture strategy further increased lipid content to 59 ± 1%, with 6.0 ± 0.3 g/L as the final concentration after the fifth cycle. The lipids produced, mainly monounsaturated fatty acids with 40% of oleic acid, offer potential as biodiesel feedstock. This lipid composition led to biodiesel properties, including cetane number, iodine value, kinematic viscosity and density that met international standards. Therefore, this research presents a promising alternative not only for WAS management but also for harnessing valuable resources, thereby establishing a basis for large-scale studies.
Subject(s)
Lipids , Sewage , Yarrowia , Yarrowia/metabolism , Lipids/chemistry , Waste Disposal, Fluid/methods , Nutrients/metabolism , Fertilizers/analysisABSTRACT
Erythritol is a polyol with a sweet taste but low energy value. Thanks to its valuable properties, as well as growing social awareness and nutritional trends, its popularity is growing rapidly. The aim of this study was to increase the effectiveness of erythritol production from glucose using new UV mutants of the yeast Yarrowia lipolytica obtained in the Wratislavia K1 strain. The ability of the new strains to biosynthesize erythritol and utilize this polyol was examined in shake-flask cultures and fed-batch processes conducted in a stirred tank reactor with a total glucose concentration of 300 and 400 g/L. The Wratislavia K1 strain produced erythritol most efficiently (97.5 g/L; 192 h) at an initial glucose concentration of 250 g/L (total: 300 g/L). New strains were assessed under such conditions, and it was noted that the highest erythritol concentration (145 g/L; 183 h) was produced by the K1UV15 strain. A significant improvement in the erythritol biosynthesis efficiency (148 g/L; 150 h) was achieved upon the increase in (NH4)2SO4 to 3.6 g/L. Further, in the culture with such a concentration of the nitrogen source and increased total glucose level (400 g/L), the K1UV15 strain produced 226 g/L of erythritol within 281 h.
Subject(s)
Erythritol , Glucose , Mutation , Yarrowia , Erythritol/metabolism , Yarrowia/metabolism , Yarrowia/genetics , Yarrowia/growth & development , Glucose/metabolism , Fermentation , Ultraviolet Rays , BioreactorsABSTRACT
The strain Gluconobacter oxydans LMG 1385 was used for the bioconversion of crude glycerol to dihydroxyacetone. The suitability of fed-batch cultures for the production of dihydroxyacetone was determined, and the influence of the pH of the culture medium and the initial concentration of glycerol on maximizing the concentration of dihydroxyacetone and on the yield and speed of obtaining dihydroxyacetone by bioconversion was examined. The feeding strategy of the substrate (crude glycerol) during the process was based on measuring the dissolved oxygen tension of the culture medium. The highest concentration of dihydroxyacetone PK = 175.8 g·L-1 and the highest yield YP/Sw = 94.3% were obtained when the initial concentration of crude glycerol was S0 = 70.0 g·L-1 and the pH of the substrate was maintained during the process at level 5.0.
Subject(s)
Batch Cell Culture Techniques , Culture Media , Dihydroxyacetone , Gluconobacter oxydans , Glycerol , Gluconobacter oxydans/metabolism , Dihydroxyacetone/metabolism , Dihydroxyacetone/biosynthesis , Glycerol/metabolism , Batch Cell Culture Techniques/methods , Culture Media/chemistry , Hydrogen-Ion Concentration , FermentationABSTRACT
BACKGROUND: Low rumen pH is proposed to be a major mechanism for low methane (CH4) emissions from sheep fed forage rape. However, it is difficult to separate this from other in vivo factors, such as rumen passage rate. The objective of this study was to determine the effect of pH alone on CH4 production in vitro using different pH buffers. Ryegrass, white clover and forage rape were incubated in vitro using three different incubation buffers with starting pH values of 5.5, 6.2 and 6.8. RESULTS: Decreasing pH reduced overall in vitro CH4 emission relative to fermented hexoses (CH4/FHex) by up to 54% and overall fermentation by 40%. pH also changed fermentation profiles where the acetate + butyrate to propionate + valerate ratio decreased when pH decreased. Within the three forages, forage rape led to the lowest CH4/FHex, but only in pH 5.5 and 6.2 buffer, and this was enhanced when the pH fell below 6. CONCLUSION: Reducing pH in vitro decreased CH4 production and overall fermentation across all forages. The lower pH reached by forage rape compared to ryegrass and white clover appears to drive the lower CH4 production relative to the extent of fermentation from forage rape compared to the other forages. © 2024 The Author(s). Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.
ABSTRACT
Adenoviruses are efficient and safe vectors for delivering target antigens and adenovirus-based vaccines have been used against a wide variety of pathogens, including tuberculosis and COVID-19. Cost-effective and scalable biomanufacturing processes are critical for the commercialization of adenovirus-vectored vaccines. Adenoviral vectors are commonly produced through the infection of batch cultures at low cell density cultures, mostly because infections at high cell densities result in reduced cell-specific virus productivity and does not improve volumetric productivity. In this study, we have investigated the feasibility of improving the volumetric productivity by infecting fed-batch cultures at high cell densities. Four commercial and one in-house developed serum-free media were first tested for supporting growth of HEK 293 cells and production of adenovirus type 5 (Ad5) in batch culture. Two best media were then selected for development of fed-batch culture to improve cell growth and virus productivity. A maximum viable cell density up to 16 × 106 cells/mL was achieved in shake flask fed-batch cultures using the selected media and commercial or in-house developed feeds. The volumetric virus productivity was improved by up to six folds, reaching 3.0 × 1010 total viral particles/mL in the fed-batch culture cultivated with the media and feeds developed in house and infected at a cell density of 5 × 106 cells/mL. Additional rounds of optimization of media and feed were required to maintain the improved titer when the fed-batch culture was scaled up in a bench scale (3 L) bioreactor. Overall, the results suggested that fed-batch culture is a simple and feasible process to significantly improve the volumetric productivity of Ad5 through optimization and balance of nutrients in culture media and feeds.
ABSTRACT
This in vitro batch culture study investigated the effects of red osier dogwood (ROD) extract supplementation on gas production (GP), dry matter disappearance (DMD), and fermentation characteristics in high forage (HF) and high grain (HG) diets with varying media pH level. The experiment was a factorial arrangement of treatments in a completely randomized design with 2 media pH (5.8 and 6.5) × 4 dose rates of ROD extract (0, 1, 3, and 5% of DM substrate). An additional treatment of monensin was added as a positive control for each pH level. The HF substrate consisted of 400 and 600 g/kg DM barley-based concentrate and barley silage, respectively, while the HG substrate contained 100 and 900 g/kg DM barley silage and barley-based concentrate, respectively. Treatments were incubated for 24 h with GP, DMD and fermentation parameters determined. No interaction was detected between the media pH level and ROD extract dose rate on GP, DMD and most of the fermentation parameters. The GP, DMD, and total volatile fatty acid (VFA) concentration were greater (P = 0.01) with media pH of 6.5 in both HF and HG diets. The GP were not affected by increasing ROD dose rate, except that GP linearly decreased in the HF (P = 0.04) and HG (P = 0.01) diets at 24 h; the DMD tended to linearly decrease at pH 6.5 (P = 0.06) for both HF and HG diets and at pH 5.8 (P = 0.02) for the HG diet. Adding ROD extract to the HF and HG diets linearly (P = 0.01) increased the acetate molar proportion at high or low media pH and consequently, the acetate to propionate (A:P) ratio linearly (P ≤ 0.04) increased. Supplementation of ROD extract to the HF diet linearly (P = 0.04) decreased the molar proportion of propionate at pH 6.5 (interaction between pH and ROD extract; P = 0.05), but had no effect on propionate proportion when added to the HG diet. Moreover, the proportion of branched-chain fatty acids linearly (P = 0.03) decreased with ROD extract supplementation at low pH (interaction, P < 0.05) for HF diet and linearly decreased (P = 0.05) at pH 6.5 for HG diet (interaction, P < 0.05). The NH3-N concentration was not affected by ROD supplementation in the HF diet but it linearly (P = 0.01) decreased with increasing dose rate in the HG diet. Methane concentration tended to linearly (P = 0.06) increase with ROD extract supplementation at high pH for HF diet and linearly increased at pH 5.8 (P = 0.06) and pH 6.5 (P = 0.02) for HG diet. These results indicate that the decreased DMD and increased A:P ratio observed with addition of ROD extract may be beneficial to HG-fed cattle to reduce the risk of rumen acidosis without negatively impacting fiber digestion.
ABSTRACT
Background: Batch cultures used for various purposes, such as expression screening and recombinant protein production in laboratories, usually have some drawbacks due to the bolus addition of carbon sources, such as glucose and buffers, that lead to overflow metabolism, decreased pH, high osmolality, low biomass yield, and low protein production. Objectives: This study aimed to overcome the problems of batch culture using the controlled release concept by a controlled porosity osmotic pump (CPOP) system. Methods: The CPOP was formulated with glucose as a carbon source feeding and sodium carbonate as a pH modifier in the core of the tablet that was coated with a semipermeable membrane containing cellulose acetate and polyethylene glycol (PEG) 400. The release rate was regulated with Eudragit L100 as a retardant agent in the core and PEG 400 as a pore-former agent in the coating membrane. Fourier-transform infrared spectroscopy (FT-IR) and scanning electron microscopy (SEM) were used to elucidate compatibility between components and release mechanism, respectively. The in-vitro release of glucose and Na2CO3 studies were performed for 24 hours in a mineral culture medium (M9). Then, the effectiveness of CPOP in the growth of Escherichia coli (E. coli BL21) as a microorganism model was evaluated. Glucose consumption, changes in medium's pH, and acetate concentration as a by-product were also monitored during the bacterial growth. Results: Fourier-transform infrared spectroscopy confirmed the compatibility between the components in the osmotic pump, and SEM elucidated the release mechanism due to in-situ delivery pores created by dissolving soluble components (PEG 400) on the coated membrane upon contact with the dissolution medium. The in-vitro release studies indicated that the osmotic pump was able to deliver glucose and sodium carbonate in a zero-order manner. The use of CPOP in E. coli (BL21) cultivation resulted in a statistically significant improvement in biomass (over 80%), maintaining the pH of the medium (above 6.8) during the exponential phase, and reducing metabolic by-product formation (acetate), compared to bolus feeding (P < 0.05). Conclusions: The use of CPOP, which is capable of controlled release of glucose as a carbon source and sodium carbonate as a pH modifier, can overcome the drawbacks of bolus feeding, such as decreased pH, increased acetate concentration, and low productivity. It has a good potential for commercialization.
ABSTRACT
Due to the rapid increase in the world's population, many developing countries are facing malnutrition problems, including famine and food insecurity. Particularly, the deficiency of protein sources becomes a serious problem for human and animal nutrition. In this context, Single Cell Proteins, could be exploited as an alternative source of unconventional proteins. The aim of the study was to investigate SCP production and composition by Cupriavidus necator under various environmental conditions, temperature and pH values. A mono-factorial approach was implemented using batch bioreactor cultures under well-controlled conditions. Results were compared in terms of bacterial growth and SCP composition (proteins, nucleic acids, amino acids and elemental formula). Complementary analyses were performed by flow cytometry to study cell morphology, membrane permeability and the presence of Poly(3-hydroxybutyrate) (PHB) production. Our data confirmed the ability of C. necator to produce high amount of proteins (69â¯%DW at 30⯰C and pH7). The results showed that temperature and pH independently impact SCP production and composition. This impact was particularly observed at the highest temperature (40⯰C) and also the lowest pH value (pH5) providing lower growth rates, cell elongation, changes in granularity and lower amounts of proteins (down to 44â¯%DW at pH5) and nucleic acids. These low percentages were related to the production of PHB production (up to 44â¯%DW at 40⯰C) which is the first report of a PHB accumulation in C. necator under nutrient unlimited conditions.
Subject(s)
Bioreactors , Cupriavidus necator , Polyesters , Temperature , Cupriavidus necator/metabolism , Cupriavidus necator/growth & development , Hydrogen-Ion Concentration , Bioreactors/microbiology , Polyesters/metabolism , Bacterial Proteins/metabolism , Hydroxybutyrates/metabolism , Prohibitins , Amino Acids/metabolism , Polyhydroxybutyrates , Dietary ProteinsABSTRACT
Tribonema minus, a promising filamentous oleaginous microalga, was cultured under different nutrient concentrations and different culture modes (fed-batch culture, two-step culture) to study the method of rapid regulation of its lipid metabolism. In contrast to many other oleaginous microalgae, T. minus did not show that nitrogen stress promoted lipid accumulation; however, sulfur deficiency promoted rapid lipid accumulation with a maximum lipid content of 54% of dry weight. Increasing the MgSO4 concentration significantly increased nitrogen uptake and biomass (10.09 g/L). Lipid productivity was significantly increased by the two-step culture using a medium with a high concentration of MgSO4 in the first step and a sulfur-free medium in the second step. In addition, it was found that the lipid content of T. minus was negatively correlated with the intracellular sulfur content when the intracellular sulfur content was below 0.6%. This study provides a new approach for industrial lipid production in T. minus.
Subject(s)
Microalgae , Stramenopiles , Stramenopiles/metabolism , Microalgae/metabolism , Biomass , Nitrogen/metabolism , LipidsABSTRACT
To improve the cell productivity of Corynebacterium glutamicum, its initial specific growth rate was improved by medium improvement using deep neural network (DNN)-assisted design with Bayesian optimization (BO) and a genetic algorithm (GA). To obtain training data for the DNN, experimental design with an orthogonal array was set up using a chemically defined basal medium (GC XII). Based on the cultivation results for the training data, specific growth rates were observed between 0.04 and 0.3/h. The resulting DNN model estimated the test data with high accuracy (R2test ≥ 0.98). According to the validation cultivation, specific growth rates in the optimal media components estimated by DNN-BO and DNN-GA increased from 0.242 to 0.355/h. Using the optimal media (UCB_3), the specific growth rate, along with other parameters, was evaluated in batch culture. The specific growth rate reached 0.371/h from 3 to 12 h, and the dry cell weight was 28.0 g/L at 22.5 h. From the cultivation, the cell yields against glucose, ammonium ion, phosphate ion, sulfate ion, potassium ion, and magnesium ion were calculated. The cell yield calculation was used to estimate the required amounts of each component, and magnesium was found to limit the cell growth. However, in the follow-up fed-batch cultivation, glucose and magnesium addition was required to achieve the high initial specific growth rate, while appropriate feeding of glucose and magnesium during cultivation resulted in maintaining the high specific growth rate, and obtaining a cell yield of 80 g/Lini.
Subject(s)
Corynebacterium glutamicum , Deep Learning , Corynebacterium glutamicum/genetics , Bayes Theorem , Magnesium , Glucose , Cell CountABSTRACT
Introduction: This study aimed to investigate the digestive function, urea utilization ability, and bacterial composition changes in rumen microbiota under high urea (5% urea in diet) over 23 days of continuous batch culture in vitro. Methods: The gas production, dry matter digestibility, and bacterial counts were determined for the continuously batch-cultured rumen fluid (CRF). The changes in fermentation parameters, NH3-N utilization efficiency, and microbial taxa were analyzed in CRF and were compared with that of fresh rumen fluid (RF), frozen rumen fluid (FRF, frozen rumen fluid at -80°C for 1 month), and the mixed rumen fluid (MRF, 3/4 RF mixed with 1/4 CRF) with in vitro rumen fermentation. Results: The results showed that the dry matter digestibility remained stable while both the microbial counts and diversity significantly decreased over the 23 days of continuous batch culture. However, the NH3-N utilization efficiency of the CRF group was significantly higher than that of RF, FRF, and MRF groups (p < 0.05), while five core genera including Succinivibrio, Prevotella, Streptococcus, F082, and Megasphaera were retained after 23 days of continuous batch culture. The NH3-N utilization efficiency was effectively improved after continuous batch culture in vitro, and Streptococcus, Succinivibrio, Clostridium_sensu_stricto_1, p.251.o5, Oxalobacter, Bacteroidales_UCG.001, and p.1088.a5_gut_group were identified to explain 75.72% of the variation in NH3-N utilization efficiency with the RandomForest model. Conclusion: Thus, core bacterial composition and function retained under high urea (5% urea in diet) over 23 days of continuous batch culture in vitro, and bacterial biomarkers for ammonia utilization were illustrated in this study. These findings might provide potential applications in improving the efficiency and safety of non-protein nitrogen utilization in ruminants.
ABSTRACT
In this study, an automatic control system is developed for the setpoint control of the cell biomass specific growth rate (SGR) in fed-batch cultivation processes. The feedback signal in the control system is obtained from the oxygen uptake rate (OUR) measurement-based SGR estimator. The OUR online measurements adapt the system controller to time-varying operating conditions. The developed approach of the PI controller adaptation is presented and discussed. The feasibility of the control system for tracking a desired biomass growth time profile is demonstrated with numerical simulations and fed-batch culture E.coli control experiments in a laboratory-scale bioreactor. The procedure was cross-validated with the open-loop digital twin SGR estimator, as well as with the adaptive control of the SGR, by tracking a desired setpoint time profile. The digital twin behavior statistically showed less of a bias when compared to SGR estimator performance. However, the adaptation-when using first principles-was outperformed 30 times by the model predictive controller in a robustness check scenario.
ABSTRACT
Spiculisporic acid (SA) is a fatty acid-type biosurfactant with one lactone ring and two carboxyl groups. It has been used in metal removers and cosmetics, because of its low propensity to cause irritation to the skin, its anti-bacterial properties, and high surface activity. In the present study, we report an effective method for producing SA by selecting a high-producing strain and investigating the effective medium components, conditions, and environments for its culture. Among the 11 kinds of Talaromyces species, T. trachyspermus NBRC 32238 showed the highest production of a crystalline substance, which was determined to be SA using NMR. The strain was able to produce SA under acidic conditions from hexoses, pentoses, and disaccharides, with glucose and sucrose serving as the most appropriate substrates. Investigation of nitrogen sources and trace metal ions revealed meat extract and FeCl3 as components that promoted SA production. Upon comparing the two types of cultures with glucose in a baffle flask or aeration bioreactor, SA production was found to be slightly higher in the flask than in the reactor. In the bioreactor culture, sucrose was found to be an appropriate substrate for SA production, as compared to glucose, because with sucrose, the lag time until the start of SA production was shortened. Finally, fed-batch culture with sucrose resulted in 60 g/L of SA, with a total yield of 0.22 g SA/g sucrose and a productivity of 6.6 g/L/day.
ABSTRACT
Abstract Food supplements have been increasingly investigated. Probiotics have several benefits for human and animal health and selenium (Se) is widely recommended against oxidative stress. In this context, the aim of this study was to develop a low-cost bioprocess to produce a functional food product comprising both probiotic and Se accumulation. Yeast cells of Saccharomyces boulardii CCT 4308 were cultivated using sugarcane molasses as substrate. Optimization studies were performed to evaluate the best medium composition for biomass production and Se-accumulation in batch and fed-batch systems. Optimized conditions were defined with a medium composed of 150 g L-1 sugarcane molasses and 12 g L-1 yeast extract, with feeding of 100 g L-1 sugarcane molasses and 100 μg mL-1 of Se incorporation after 4 h and 10 h of fermentation, respectively, during 48 h in STR (stirred tank reactor). Best biomass production reached 14.52 g L-1 with 3.20 mg Se g-1 biomass at 12 h. Process optimization led to 4.82-fold increase in biomass production compared to initial condition. A final Se-enriched S. boulardii CCT 4308 biomass was obtained, which is comparable to commercial products. An alternative probiotic yeast biomass was efficiently produced as a new food-form of Se supplement in a sustainable process using an inexpensive agro-industrial residue.
Subject(s)
Selenium , Molasses , Biomass , Probiotics , Saccharomyces boulardiiABSTRACT
BACKGROUND: Lawsonia intracellularis remains a problem for the swine industry worldwide. Previously, we designed and obtained a vaccine candidate against this pathogen based on the chimeric proteins: OMP1c, OMP2c, and INVASc. These proteins formed inclusion bodies when expressed in E. coli, which induced humoral and cellular immune responses in vaccinated pigs. Also, protection was demonstrated after the challenge. In this study, we established a production process to increase the yields of the three antigens as a vaccine candidate. RESULTS: Batch and fed-batch fermentations were evaluated in different culture conditions using a 2 L bioreactor. A fed-batch culture with a modified Terrific broth medium containing glucose instead of glycerol, and induced with 0.75 mM IPTG at 8 h of culture (11 g/L of biomass) raised the volumetric yield to 627.1 mg/L. Under these culture conditions, plasmid-bearing cells increased by 10% at the induction time. High efficiency in cell disruption was obtained at passage six using a high-pressure homogenizer and a bead mill. The total antigen recovery was 64% (400 mg/L), with a purity degree of 70%. The antigens retained their immunogenicity in pigs, inducing high antibody titers. CONCLUSIONS: Considering that the antigen production process allowed an increment of more than 70-fold, this methodology constitutes a crucial step in the production of this vaccine candidate against L. intracellularis.
Subject(s)
Animals , Swine Diseases/immunology , Bacterial Vaccines/immunology , Lawsonia Bacteria/immunology , Desulfovibrionaceae Infections/prevention & control , Swine , Swine Diseases/prevention & control , Bacterial Vaccines/administration & dosage , Vaccines, Synthetic , Cell Survival , Vaccination , Fermentation , Batch Cell Culture Techniques , ImmunityABSTRACT
Background The production of biofuels from renewable energy sources is one of the most important issues in biotechnology today. The process is known to generate various by-products, for example glycerol that is obtained in the making of biodiesel from rapeseed oil. Crude glycerol may be utilized in many ways, including microbial conversion to 1,3-propanediol. The main drawback of that technology is the use of high concentrations of glycerol, which inhibits the growth of bacterial cells. Results This study investigated the impact of crude glycerol on Clostridium butyricum DSP1 and its ability to adapt to an environment of high osmotic pressure. It was found that a crude glycerol concentration of up to 70 g/L did not have an inhibitory effect on C. butyricum DSP1. Adaptation procedures involving the passage of metabolically active biomass from a fermentation medium with a lower concentration of crude glycerol to one with a greater substrate concentration allowed breaking the barrier of high osmotic pressure (150 g/L crude glycerol) and receiving a 1,3-PD concentration of 74 g/L in a batch culture operation. The work looked into intracellular modifications shown by proteomic profiling in order to explain the mechanisms underlying the response and adaptation of bacterial cells exposed to unfavorable environmental conditions. Conclusions This study of the effect of glycerol on the growth and metabolism of C. butyricum DSP1 demonstrated that the maximum substrate concentrations that do not inhibit the metabolic activity of bacterial cells are 90 g/L and 70 g/L for pure and crude glycerol, respectively.
Subject(s)
Adaptation, Physiological , Clostridium butyricum/growth & development , Clostridium butyricum/metabolism , Glycerol/metabolism , Osmotic Pressure , Propylene Glycols , Stress, Physiological , Proteins/analysis , Environment , Biofuels , Fermentation , Batch Cell Culture Techniques , Glycerol/analysisABSTRACT
The production of lactic acid from date juice by
Subject(s)
Batch Cell Culture Techniques , Lactic Acid/metabolism , Lacticaseibacillus casei/metabolism , Lacticaseibacillus rhamnosus/metabolism , Phoeniceae/metabolism , Fermentation , Plant Extracts/metabolismABSTRACT
Bacillus amyloliquefaciens fmb50 produces a high yield of surfactin, a lipopeptide-type biosurfactant that has been widely studied and has potential applications in many fields. A foam overflowing culture has been successfully used in the combined production-enrichment fermentation of surfactin. In this study, the agitation and aeration rates were found to have relationships with foam formation and surfactin enrichment. A maximum surfactin concentration of 4.7 g/l of foam was obtained after 21 h of culture with an agitation rate of 150 rpm and an aeration rate of 1 vvm in fed-batch culture. By controlling the foam overflow rate (f out) of a fed-batch culture, surfactin concentration in the foam was continuously maintained above 4 g/l.
Bacillus amyloliquefaciens fmb50 produce gran cantidad de surfactina, un biosurfactante de tipo lipopeptídico que ha sido objeto de estudios pormenorizados y tiene aplicaciones en muchos campos. El cultivo en espuma desbordante se ha utilizado con éxito en la fermentación combinada de producción-enriquecimiento de surfactina. En este estudio, se halló que las tasas de aireación y agitación tienen relación con la formación de espuma y el enriquecimiento de la surfactina. Se obtuvo una concentración máxima de surfactina de 4,7 g/l de espuma después de 21 h de cultivo con una tasa de agitación de 150 rpm y una tasa de aireación de 1 vvm en un cultivo alimentado (fed-batch). Al controlar la tasa de espuma desbordante (f out) de un cultivo fed-batch, la concentración de surfactina en la espuma se mantuvo continua por encima de 4 g/l.