Optimization of the ellagic acid synthesis process at the bioreactor level using non-conventional yeasts

Abstract Ellagic acid (EA) is a phenolic biomolecule. For its biosynthesis, a source of ellagitannins is required, such as strawberries and yeasts, as precursors of the tannase and ß-glucosidase enzymes responsible for hydrolysis of ellagitannins. Two experimental mixture designs were applied., varying the yeast concentration and the number of ellagitannins in the culture medium, evaluating the enzymatic activity and ellagic acid biosynthesis. Aiming to find the optimal compositions of the non-conventional yeasts assessed in the research to biosynthesize ellagic acid feasibly and efficiently using a response surface performing the statistical analysis in the StatGraphics® program for obtaining a higher yield and optimizing the ellagic acid synthesis process, the results indicate that the strains Candida parapsilosis ITM LB33 and Debaryomyces hansenii ISA 1510 have a positive effect on the synthesis of ellagic acid, since as its concentration increases in the mixture the concentration of ellagic acid in the medium also increases; on the other hand, the addition of Candida utilis ITM LB02 causes a negative effect, resulting in the compositions of 0.516876, 0.483124 and 2.58687E-9 respectively, for a treatment under the same conditions, an optimal value of ellagic acid production would be obtained. With an approximate value of 7.33036 mg/mL

The Impact of Different Powdered Mineral Materials on Selected Properties of Aerobic Granular Sludge.

This study aimed to evaluate and compare the physical, chemical and biological properties of aerobic granular sludge from reactors with the addition of different powdered mineral materials. These properties have a significant impact on the efficiency of systems in which the biomass in granular form is used. Four identical granular sequencing batch reactors (GSBRs) were adopted for the research performed on a laboratory scale (R1-control reactor; R2, R3 and R4-with materials, PK, PG and PL respectively). The results indicate that the addition of powdered mineral materials improved the properties of biomass in reactors. The SVI5/SVI30 ratio values were significantly lower in the reactors with added materials (approx. 1.3 ± 0.3). The mean values of the sludge volume index at 30 min were the lowest in the R2 (39.8 ± 8.6 mL/g) and R4 (32.8 ± 10.7 mL/g) reactors. The settling velocity of biomass was the highest in the R2 reactor (15.4 ± 6.1 m/h). In the early days of the study, the highest extracellular polymeric substances (EPS) content was found in the biomass from the reactors to which the materials with higher Ca and Mg content were added (380.18-598.30 mg/g MLVSS). The rate of specific oxygen uptake (SOUR) by biomass indicated an insufficient biomass content in the R1 reactor-to 7.85 mg O2/(g MLVSS∙h)-while in the reactors with materials, the SOUR values were at the higher levels.

Feasibility of anaerobic packed and structured-bed reactors for sulfamethoxazole and ciprofloxacin removal from domestic sewage.

This study assessed the applicability of fixed bed bioreactors in two configurations - anaerobic structured bed reactor (ASBR) and anaerobic packed bed reactor (APBR) - in the removal of Sulfamethoxazole (SMX) and Ciprofloxacin (CIP), two antibiotics frequently detected in sanitary sewage. The problem of these pharmaceuticals as emerging contaminants in conventional sewage treatment systems is mainly because they encourage the development and spread of resistance genes in bacteria. Both reactors had similar performances, and the antibiotics were highly removed - APBR: 85 ±â€¯10% for SMX and 81 ±â€¯16% for CIP; ASBR: 83 ±â€¯12% for SMX and 81 ±â€¯15% for CIP. The ASBR showed to be potentially more feasible in operating and economic terms compared to the APBR, as the former presents a smaller amount of support material in the bed. SMX was completely biotransformed, while the influence of the sorption mechanism was observed for CIP, as its presence was detected sorbed onto biomass throughout the reaction bed of the reactors, with a partition coefficient (log KD) of around 2.8 L·kg-1TSS. The degradation kinetics of the pharmaceuticals were fitted using a first-order kinetic model, whereby the reactors behaved as plug flow ones, indicating the possibility of optimizing the operation for a hydraulic retention time of 6 h. The removal kinetics was more favorable for CIP (higher apparent constant kinetic - kCIPapp > kSMXapp), since its biodegradation is linked to the biomass, which is more concentrated in the bed bottom layer. The experimental results showed the potential of anaerobic fixed bed reactors in removing environmentally relevant concentrations of SMX and CIP found in sewage.

Mesenchymal stromal cell therapy: progress in manufacturing and assessments of potency.

Mesenchymal stromal cell (MSC) therapies have been pursued for a broad spectrum of indications but mixed reports on clinical efficacy have given rise to some degree of skepticism regarding the effectiveness of this approach. However, recent reports of successful clinical outcomes and regulatory approvals for graft-versus-host disease, Crohn's disease and critical limb ischemia have prompted a shift in this perspective. With hundreds of clinical trials involving MSCs currently underway and an increasing demand for large-scale manufacturing protocols, there is a critical need to develop standards that can be applied to processing methods and to establish consensus assays for both MSC processing control and MSC product release. Reference materials and validated, uniformly applied tests for quality control of MSC products are needed. Here, we review recent developments in MSC manufacturing technologies, release testing and potency assays. We conclude that, although MSCs hold considerable promise clinically, economies of scale have yet to be achieved although numerous bioreactor technologies for scalable production of MSCs exist. Additionally, rigorous disease-specific product testing and comprehensive understanding of mechanisms of action, which are linked to relevant process and product release potency assays, will be required to ensure that these therapies continue to be successful.

Biodiversity and technological potential of lactic acid bacteria isolated from spontaneously fermented amaranth sourdough.

UNLABELLED: Spontaneous fermented sourdoughs prepared from amaranth flour were investigated for the presence of autochthonous lactic acid bacteria (LAB) predominating microbiota. The doughs were fermented with daily backslopping on a laboratory scale at 30°C for 10 days. LAB counts ranged from 2·60 to 8·54 log CFU g(-1) with a pH declined from 6·2 to 3·8 throughout fermentation. The combined use of randomly amplified polymorphic DNA (RAPD)-PCR analysis and sequence analysis of 16S rRNA was applied for LAB intraspecies differentiation and taxonomic identification, respectively. Enterococcus, Pediococcus and Lactobacillus species were present in amaranth sourdoughs (AS). After the first refreshment step, Lactobacillus plantarum dominated AS until the end of fermentation. In coincidence, when DGGE analysis was performed, the occurrence of a progressive change in bacterial communities allowed the selection of Lact. plantarum as a dominant species. Moreover, technological, functional and safety characteristics of representative RAPD-biotypes were investigated. Lact. plantarum CRL1898 was selected as a potential candidate for gluten-free amaranth sourdough starter. SIGNIFICANCE AND IMPACT OF THE STUDY: Nowadays, there is an increasing interest in ancient noncereal gluten-free (GF) crops such as amaranth, due to their reported nutritional and health benefits. However, the use of these grains is still limited to traditional foods and bread making processes that are not yet well standardized. Results on the dynamics of autochthonous lactic acid bacteria (LAB) microbiota during laboratory spontaneous amaranth sourdoughs (AS) fermentation will contribute to overcome challenges for GF-fermented products development. In addition, knowledge about LAB diversity involving Enterococcus, Pediococcus and Lactobacillus species, with Lactobacillus plantarum predominating during AS fermentation, and their technological and functional properties provides the basis for the selection of autochthonous strains as starters cultures for novel gluten-free bakery products with enhanced nutritional, sensory and/or safety quality.

Nitrite reduction and methanogenesis in a single-stage UASB reactor.

In this study, nitrite reduction and methanogenesis in a single-stage upflow anaerobic sludge blanket (UASB) reactor was investigated, using high-strength synthetic domestic wastewater as substrate. To assess long-term effects and evaluate the mechanisms that allow successful nitrite reduction and methanogenesis in a single-stage UASB, sludge was exposed to relatively high nitrite loading rates (315 ± 13 mgNO(2)(-)-N/(l.d)), using a chemical oxygen demand (COD) to nitrogen ratio of 18 gCOD/gNO(2)(-)-N, and an organic loading rate of 5.4 ± 0.2 gCOD/(l.d). In parallel, the effects of sludge morphology on methanogenesis inhibition were studied by performing short-term batch activity tests at different COD/NO(2)(-)-N ratios with anaerobic sludge samples. In long-term tests, denitrification was practically complete and COD removal efficiency did not change significantly after nitrite addition. Furthermore, methane production only decreased by 13%, agreeing with the reducing equivalents requirement for complete NO(2)(-) reduction to N2. Apparently, the spatial separation of denitrification and methanogenesis zones inside the UASB reactor allowed nitrite reduction and methanogenesis to occur at the same moment. Batch tests showed that granules seem to protect methanogens from nitrite inhibition, probably due to transport limitations. Combined COD and N removal via nitrite in a single-stage UASB reactor could be a feasible technology to treat high-strength domestic wastewater.

Treatment of domestic wastewater with an anaerobic ceramic membrane bioreactor (AnCMBR).

In this study, a ceramic membrane with a pore size of 80 nm was incorporated into an anaerobic membrane bioreactor for excellent stability and integrity. Chemical oxygen demand (COD) removal efficiencies by biodegradation reached 78.6 ± 6.0% with mixed liquor suspended solids (MLSS) of 12.8 ± 1.2 g/L. Even though the total methane generated was 0.3 ± 0.03 L/g CODutilized, around 67.4% of it dissolved in permeate and was lost beyond collection. As a result, dissolved methane was 2.7 times of the theoretical saturating concentration calculated from Henry's law. When transmembrane pressure (TMP) of the ceramic membrane reached 30 kPa after 25.3 d, 95.2% of the total resistance was attributed to the cake layer, which made it the major contributor to membrane fouling. Compared to the mixed liquor, cake layer was rich in colloids and soluble products that could bind the solids to form a dense cake layer. The Methanosarcinaceae family preferred to attach to the ceramic membranes.

In vivo bioreactors for mandibular reconstruction.

Large mandibular defects are difficult to reconstruct with good functional and aesthetic outcomes because of the complex geometry of craniofacial bone. While the current gold standard is free tissue flap transfer, this treatment is limited in fidelity by the shape of the harvested tissue and can result in significant donor site morbidity. To address these problems, in vivo bioreactors have been explored as an approach to generate autologous prefabricated tissue flaps. These bioreactors are implanted in an ectopic site in the body, where ossified tissue grows into the bioreactor in predefined geometries and local vessels are recruited to vascularize the developing construct. The prefabricated flap can then be harvested with vessels and transferred to a mandibular defect for optimal reconstruction. The objective of this review article is to introduce the concept of the in vivo bioreactor, describe important preclinical models in the field, summarize the human cases that have been reported through this strategy, and offer future directions for this exciting approach.

Evolution of filamentous bacteria during urban wastewater treatment by MBR.

Evolution of filamentous bacteria in two full-scale experimental MBR systems (microfiltration and ultrafiltration) was studied during two years. Sludge Retention Time (SRT) and Hydraulic Retention Time (HRT) were modified and acted as variables, together with temperature and variation in loading. With SRT values between 20 and 35 d and HRT between 31 and 40 h, both MBR systems presented a high density of filamentous bacteria, according to the Filamentous Index (FI) and Simplified Technique of Filamentous Count (STFC). Highest density was achieved when contaminant loads were high and temperature was low. However, the elevated presence of filamentous bacteria did not affect the quality of effluent or the permeability of the membranes. Nocardioform bacteria showed a high degree of adaptation to the characteristics of the system. Predominance of Nocardioforms gave rise to isolated episodes of massive growth at temperatures between 15 and 20°C, which in turn caused episodes of intense foaming whose most significant consequence was a loss in biomass, leading to a slight increase in transmembrane pressure. In the light of these results, FI and STFC should not be considered as suitable tools for predicting operational problems deriving from filamentous bacteria in MBR systems, which could be prevented through identification.

[Cultivation of Panax ginseng adventitious roots in bubble bioreactors].

OBJECTIVE: To study cultivation of Panax ginseng adventitious roots in bubble bioreactors. METHOD: The adventitious roots were obtained through tissue culture different types of bioreactors. The contents of ginsenosides Re, Rb1 and Rg1 were determined by HPLC while the contents of polysaccharides were determined by ultraviolet spectrophotometry. RESULT: The results showed that of the three types tested, the most efficient bioreactor for cultivation of the ginseng adventitious roots was the cone-type bioreactors (with the 120 degrees ), in which, the growth curve of adventitious roots was S-shaped. The maximum biomass was obtained on the 40th day, with the fresh weight, dry weight and growth rate reaching the maximum, which were 113.15 g, 9.62 g and 63.13 times respectively, and the concomitant contents of polysaccharide and ginsenoside were 2.73% and 2.25 mg x g(-1). CONCLUSION: The results showed that the most efficient bioreactor for cultivation of the ginseng adventitious roots was the cone-type bioreactors (with the 120 degrees). These results provide a theoretical reference for developing an efficient production process of active metabolites of ginseng in the scale-up cultivation.

Bioreactor design for continuous dark fermentative hydrogen production.

Dark fermentative H2 production (DFHP) has received increasing attention in recent years due to its high H2 production rate (HPR) as well as the versatility of the substrates used in the process. For most studies in this field, batch reactors have been applied due to their simple operation and efficient control; however, continuous DFHP operation is necessary from economical and practical points of view. Continuous systems can be classified into two categories, suspended and immobilized bioreactors, according to the life forms of H2 producing bacteria (HPB) used in the reactor. This paper reviews operational parameters for bioreactor design including pH, temperature, hydraulic retention time (HRT), and H2 partial pressure. Also, in this review, various bioreactor configurations and performance parameters including H2 yield (HY), HPR, and specific H2 production rate (SHPR) are evaluated and presented.

Performance of a MBR for the treatment of blackwater.

The paper describes the experience gained in operating a membrane bioreactor (MBR) for the treatment of blackwater. Beside a complete characterization of blackwater, operational conditions and removal efficiency concerning parameters such as COD, BOD(5), nitrogen and phosphorus as well as microbiological parameters were determined. Furthermore the membrane performance was investigated. The results show that in blackwater treatment nitrogen removal is limited in the biological process, because of the blackwater matrix (BOD(5):TKN=1.1:1.0). Blackwater contains a high fraction of soluble, inert COD, which is not degradable by biological operation, only. Phosphorus elimination was negligible, probably induced by precipitation of cellular phosphorus. Although the released permeate was free of the fecal indicators E. coli and streptococcus and met guideline values, a direct reuse as service water is not recommended due to the yellowish coloration.

Mesophilic anaerobic digestion: first option for waste treatment in tropical regions.

Rural India derives its energy needs for cooking and heating through the use of fuel wood and for lighting and agricultural operations through kerosene and diesel. Use of fuel wood has aggravated the problem of de-forestation, while availability of kerosene and diesel cannot be guaranteed due to corrupt practices in the public distribution systems. In contrast, urban India derives its energy needs through LPG cylinders, petrol, and electricity. However, their cost and uncertainty rendered them beyond the reach of lower income population. This scenario is more or less true with many developing countries. To meet these objectives, biogas generation from biodegradable waste using anaerobic digestion (AD) appears to be a sustainable avenue as it could be used for (a) water and space heating of farmhouses, animal shelters, (b) generating steam for food processing plants, and (c) electricity generation, in addition to reducing the pollution/hazard potential of these wastes. Many of the underdeveloped and developing countries are in the temperate zone and thus mesophilic AD could provide a desired pathway to achieve a long delayed need of energy for comfortable living, farming, and industrial operations. Efforts made in this direction are reviewed in the present article.

Comparison of bioreactors with different kinds of submerged packed beds for domestic wastewater treatment.

The performance of aerobic submerged packed bed reactors was studied for the treatment of domestic wastewater using different kinds of packing materials with high specific areas (760-1,200 m(2)/m(3)). The tested materials were ceramic spheres, crushed tezontle, grains of high density polyethylene (HDPE), of low density polyethylene (LDPE) and of polypropylene (PP), cubes of polyurethane (PU) and polyethylene tape (SESSIL). The bioreactors were operated in continuous regime, applying organic loads in the range of 0.8-6.0 g COD.m(-2).d(-1). The obtained specific COD removal rates were very similar in all the reactors when they were operated at organic loads up to 2.0 g COD.m(-2).d(-1), after which differences in effectiveness appeared and the best results were determined in the reactors with SESSIL, LDPE and PU. Very low TSS, O&G and turbidity were obtained in all the effluents. The NH(3)-N and TN removals were dependent on the dissolved oxygen (DO) concentration and the removals at DO of 5 mg/l were 84-99% and 61-74% respectively. The best removals were determined in the reactors with PU, SESSIL and LDPE. The reactor with tezontle had also a good performance when operated with loads up to 1.0 g TN.m(-2).d(-1). The best phosphate removals (38-49%) were obtained in the reactors with PU, tezontle, ceramic sheres and SESSIL.

Fermentative hydrogen production in packed-bed and packing-free upflow reactors.

Fermentative hydrogen production from a synthetic wastewater containing 10 g/L of sucrose was studied in two upflow reactors at 26 degrees C for 400 days. One reactor was filled with packing rings (RP) and the other was packing free (RF). The effect of hydraulic retention time (HRT) from 2 h to 24 h was investigated. Results showed that, under steady state, the hydrogen production rate significantly increased from 0.63 L/L/d to 5.35 L/L/d in the RF when HRT decreased from 24 h to 2 h; the corresponding rates were 0.56 L/L/d to 6.17 L/L/d for the RP. In the RF, the hydrogen yield increased from 0.96 mol/mol-sucrose at 24 h of HRT to the maximum of 1.10 mol/mol-sucrose at 8 h of HRT, and then decreased to 0.68 mol/mol-sucrose at 2 h. In the RP, the yield increased from 0.86 mol/mol-sucrose at 24 h of HRT to the maximum of 1.22 mol/mol-sucrose at 14 h of HRT, and then decreased to 0.78 mol/mol-sucrose at 2 h. Overall, the reactor with packing was more effective than the one free of packing. In both reactors, sludge agglutinated into granules. The microbial community of granular sludge in RP was investigated using 16S rDNA based techniques. The distribution of bacterial cells and extracellular polysaccharides in hydrogen-producing granules was investigated by fluorescence-based techniques. Results indicated that most of the N-acetyl-galactosamine/galactose-containing extracellular polysaccharides were distributed on the outer layer of the granules with a filamentous structure.

Performance of an internal-loop airlift bioreactor for treatment of hexane-contaminated air.

Hexane is a toxic volatile organic compound that is quite abundant in gas emissions from chemical industries and printing press and painting centers, and it is necessary to treat these airstreams before they discharge into the atmosphere. This article presents a treatment for hexane-contaminated air in steady-state conditions using an internal-loop airlift bioreactor inoculated with a Pseudomonas aeruginosa strain. Bioprocesses were conducted at 20-mL/min, a load of 1.26 gm3 of C6H14, and a temperature of 28 degrees C. The results of hexane removal efficiencies were presented as a function of the inoculum size (approx 0.07 and 0.2 g/L) and cell reuse. Bioprocess monitoring comprises quantification of the biomass, the surface tension of the medium, and the hexane concentration in the fermentation medium as well as in the inlet and outlet airstreams. The steady-state results suggest that the variation in inoculum size from 0.07 to 0.2 g/L promotes hexane abatement from the influent from 65 to 85%, respectively. Total hydrocarbon removal from the waste gas was achieved during experiments conducted using reused cells at an initial microbial concentration of 0.2 g/L.

Modeling and simulation of oxygen-limited partial nitritation in a membrane-assisted bioreactor (MBR).

Combination of a partial nitritation process and an anaerobic ammonium oxidation process for the treatment of sludge reject water has some general cost-efficient advantages compared to nitrification-denitrification. The integrated process features two-stage autotrophic conversion of ammonium via nitrite to dinitrogen gas with lower demand for oxygen and no external carbon requirement. A nitrifying membrane-assisted bioreactor (MBR) for the treatment of sludge reject water was operated under continuous aeration at low dissolved oxygen (DO) concentrations with the purpose of generating nitrite accumulation. Microfiltration was applied to allow a high sludge retention time (SRT), resulting in a stable partial nitritation process. During start-up of the MBR, oxygen-limited conditions were induced by increasing the ammonium loading rate and decreasing the oxygen transfer. At a loading rate of 0.9 kg N m(-3) d(-1) and an oxygen concentration below 0.1 mg DO L(-1), conversion to nitrite was close to 50% of the incoming ammonium, thereby yielding an optimal effluent within the stoichiometric requirements for subsequent anaerobic ammonium oxidation. A mathematical model for ammonium oxidation to nitrite and nitrite oxidation to nitrate was developed to describe the oxygen-limited partial nitritation process within the MBR. The model was calibrated with in situ determinations of kinetic parameters for microbial growth, reflecting the intrinsic characteristics of the ammonium oxidizing growth system at limited oxygen availability and high sludge age. The oxygen transfer coefficient (K(L)a) and the ammonium-loading rate were shown to be the appropriate operational variables to describe the experimental data accurately. The validated model was used for further steady state simulation under different operational conditions of hydraulic retention time (HRT), K(L)a, temperature and SRT, with the intention to support optimized process design. Simulation results indicated that stable nitrite production from sludge reject water was feasible with this process even at a relatively low temperature of 20 degrees C with HRT down to 0.25 days.

A survey of computational and physical methods applied to solid-state fermentation.

During the last decade, significant effort has been made to apply computational and physical methods to solid-state fermentation (SSF). This had positive impact both on our understanding of the basic principles underlying this old technology, and on the latest progress made in industrial bioengineering. Guidelines on bioreactor design and operation including scale-up, new methods for biomonitoring and advanced control strategies are among the most important outcomes of practical use. Nevertheless, there still is a lack of experimental data, which hampers parameter identification and thus broader use of mathematical modeling. More attention should therefore be paid to combining and concentrating modern physical techniques and computational approaches in order to allow better model validation and thus further progress in rational bioengineering of SSF.

Comparison between a submerged membrane bioreactor and a conventional activated sludge system on treating ammonia-bearing inorganic wastewater.

A submerged membrane bioreactor (SMBR) and a conventional activated sludge system (CAS) were compared in parallel over a period of 210 days on treating synthetic ammonia-bearing inorganic wastewater under similar conditions. Except for a short period of pH control failure, almost complete conversion of NH(4)(+)?N to NO(3)(-)?N was constantly achieved over an NH(4)(+)?N concentration range from 180 to 1300mgl(-1) at a hydraulic retention time (HRT) of 24h in the SMBR, compared to an average conversion ratio of 95.0% in the CAS. Scanning electron micrographs (SEMs) demonstrated the accumulation of extracellular polymeric substances (EPSs) in the SMBR. Ubiquinone-8 (UQ-8), followed by UQ-10, UQ-7 and UQ-9, was the dominant ubiquinone in both the systems. The dominant menaquinone in the SMBR was menaquinone-6 (MK-6), while that in the CAS was MK-7, indicating that some differences existed between the two systems in terms of microbial community structure. Soluble microbial products (SMPs) tended to accumulate, and then biodegrade in SMBR.

Production of L-phenylacetyl carbinol by immobilized cells of Saccharomyces cerevisiae.

Conversion of benzaldehyde to L-phenylacetyl carbinol (L-PAC) was achieved with immobilized, growing cells of Saccharomyces cerevisiae in different reactors. Product formation increased (31%) with the subsequent initial reuses of the entrapped cells. Biomass production and PAC formation depleted (40 and 57%, respectively) after 4-5 continuous growth and biotransformation cycles. With the regeneration of the biocatalysts, catalytic activity of the cells was resumed. The highest yields were in a stirred tank reactor (29 g PAC) from 77 g benzeldehyde with 14 repeated uses of entrapped cells.