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The influences of the pH-gradient between the feed and stripping phases and of carrier concentration inside the membrane phase on the efficiency of facilitated pertraction of folic acid with Amberlite LA-2 in pseudosteady-state mode have been analyzed. The experiments have been carried out using a U-shaped pertraction cell which allowed to easily maintaining the liquid membrane between the two aqueous phases. On the basis of the experimental data and theoretical investigation on pseudosteady-state regime, two models describing the acid accumulation inside the liquid membrane by means of the permeability factor have been developed for two pH-values of feed phases (5.2 and 3, respectively). The proposed models offer good concordance with the experimental data and can be useful for facilitated pertraction optimization. [ABSTRACT FROM AUTHOR]

The complexity of downstream processes for biosynthetic products constitutes a particularity of industrial biotechnologies, especially because of the biosynthetic product high dilution in fermentation broth, their chemical and thermal liability and the presence of secondary products. For these reasons, new separation techniques have been developed and applied to bioseparations. Among them, reactive extraction, pertraction (extraction and transport through liquid membranes) and direct extraction from broths have considerable potential and are required for the further development of many biotechnologies. This review is structured on two parts and presents our original results of the studies on the separation of some biosynthetic products (antibiotics, carboxylic acids, amino acids, alcohols) by reactive extraction in the first part, and by pertraction and direct extraction from broths without biomass filtration in the second. For all the analyzed cases, these extraction techniques simplify the technologies by reducing material and energy consumption, by avoiding product inhibition, by increasing the separation selectivity, therefore decreasing the overall cost of the product.

The mixing time for bioreactors depends mainly on the rheoiogicai properties of the broths, the biomass concentration and morphology, mixing system characteristics and fermentation conditions. For quantifying the influence of these factors on the mixing efficiency for stirred bioreactors, aerated broths of bacteria (P. shermanii), yeasts (S. cerevisiae) and fungi (P. chrysogenum, free mycelia and mycelial aggregates) of different concentrations have been investigated using a laboratory bioreactor with a double turbine impeller. The experimental data indicated that the influence of the rotation speed, aeration rate and stirrer positions on the mixing intensity strongly differ from one system to another and must be correlated with the microorganism characteristics, namely: the biomass concentration and morphology. Moreover, compared with non-aerated broths, variations of the mixing time with the considered parameters are very different, due to the complex flow mechanism of gas-liquid dispersions. By means of the experimental data and using a multiregression analysis method some mathematical correlations for the mixing time of the general form: tm = a1*Cx2+a2*Cx+a3*IgVa+a4-N2+a5-N+a6/a7*L2+a8*L+a9 were established. The proposed equations offer good agreement with the experiments, the average deviation being ±6.7% - ±9.4 and are adequate for the flow regime Re < 25,000.

The second part of this review presents our original results on the separation of some biosynthetic products (antibiotics, carboxylic acids, alcohols) by pertraction and direct extraction from broths without biomass filtration. For the analyzed systems, the experimental conditions required for reaching maximum separation efficiency and the mathematical models describing the process have been established. For all the studied cases, these extraction techniques simplify the technologies and reduce the overall cost of the product.

Facilitated pertraction was applied for the selective separation of citric, maleic and succinic acids from a mixture obtained by citric fermentation. The pertraction equipment included a U-shaped cell containing 1,2-dichloro-ethane as the liquid membrane and Amberlite LA-2 as the carrier. The experimental data indicated that maleic and succinic acids can be initially selectively separated from citric acid, followed by the selectively separation of maleic acid from succinic acid. Using statistical analysis and a second order factorial experiment, two mathematical correlations describing the influence of the main process variables on pertraction selectivity were established. For both extraction systems, the considered variables controlled the extraction process to an extent of 92.9-99.9%, the carrier concentration inside the liquid membrane exhibiting the most important influence.

The study of rutin extraction from autochthonous hardhay flowers (Hyperici herba) and underbrush fruits (Hippophaes fructus) with methanol, ethanol and 1-propanol at different concentrations, temperatures and duration indicated that the most efficient solvent was 1-propanol, the optimum concentration being determined by the amount and type of natural compounds from the two plants.Using statistical analysis and a factorial second order experiment two mathematical correlations between the degree of rutin extraction and the main parameters influencing the process (1-propanol concentration temperature, duration) were established. For both extraction systems, the considered variables control the extraction process to an extent of 99.2 99.5%, the solvent concentration exhibiting the most important influence especially for rutin extraction from underbrush fruits (Hippophaes fructus).

The aim of these experiments is to study the oxygen mass transfer rate by means of the mass transfer coefficient, for a stirred bioreactor and different fermentation broths, using a large domain of operating variables. For quantifying the effects of the considered factors (concentration and morphology of the biomass, specific power input, superficial air velocity surface aeration) on ka, the experiments were carried out for non-respirating biomass suspensions of Propionibacterium shermanii Saccharomyces cerevisiae and Penicillium chrysogenum, mycelial aggregates (pellets) and free mycelia morphological structures.

Using the experimental data on the individual physical and reactive extraction of 6APA with 1,2-dichlororethane, D2EHPA and Amberlite LA-2 together with data on the influence of the pH value of the aqueous phase and the Amberlite LA-2 concentration on the selective separation of 6APA, PG and PAA from a mixture, a process flow sheet for the fractionation of the solution obtained by the enzymatic hydrolysis of PG has been elaborated and applied. Thus, at pH=10, 6APA was selectively separated by four extraction stages, the overall extraction degree being 98.8%. PG was selectively extracted from the raffinate at pH=6 by three extraction stages, adjusting the molar concentration of Amberlite LA-2 to the value of the PG molar concentration for each extraction stage. The antibiotic extraction degree was 99.6%.

This paper presents the experiments on mixing efficiency for aerated media for a laboratory stirred bioreactor with a double turbine impeller. The effects of stirrer rotation speed, air volumetric flow rate and stirrer position on the shaft on mixing time for aerated water and simulated broths (CMCNa solutions) were analyzed. Compared to non-aerated broths, the results indicated that the variation of mixing time with the considered parameters is very different, due to the complex flow mechanism of the gas-liquid dispersion, a mechanism which is changed by changing the broth properties or fermentation conditions. Using the Statistics Toolbox of MATLAB some correlations between the mixing time and rotation speed, air volumetric flow rate and stirrer position on the shaft were established. The proposed equations agree well with the experiments, the average deviation being ±9.02%.

In this paper the mixing distribution intos the non-aerated simulated and S. cerevisiae broths for a stirred bioreactors are studied, being recorded significant differences between the influences of viscosity, solid phase and rotation speed on the stagnant region position for the two analyzed broths. [ABSTRACT FROM AUTHOR]

In this paper the original studies on mechanism, influencing factors and kinetic of reactive extraction of vitamin B9 with Amberlite LA-2 dissolved in three different solvents are presented. [ABSTRACT FROM AUTHOR]

Muconic acid is a six-carbon dicarboxylic acid with conjugated double bonds that finds extensive use in the food (additive), chemical (production of adipic acid, monomer for functional resins and bio-plastics), and pharmaceutical sectors. The biosynthesis of muconic acid has been the subject of recent industrial and scientific attention. However, because of its low concentration in aqueous solutions and high purity requirement, downstream separation presents a significant problem. Artificial Neural Networks and Differential Evolution were used to optimize process parameters for the recovery of muconic acid from aqueous streams in a system with n-heptane as an organic diluent and ionic liquids as extractants. The system using 120 g/L tri-hexyl-tetra-decyl-phosphonium decanoate dissolved in n-heptane, pH of the aqueous phase 3, 20 min contact time, and 45 °C temperature assured a muconic acid extraction efficiency of 99,24 %. Low stripping efficiency compared to extraction efficiency was observed for the optimum conditions on the extraction step (120 g/L ionic liquids dissolved in heptane). However, re-extraction efficiencies obtained for the recycled organic phase in three consecutive stages were close to the first extraction stage. The mechanism analysis proved that the analysed phosphonium ionic liquids (PILSs) extracts only undissociated molecules of muconic acid through H-bonding., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

Known as the degenerative disease of the knee with the highest prevalence, knee osteoarthritis (KOA) is characterized by a gradual destructive mechanism that, in severe cases, can provoke the need for total knee substitution. As the disease progresses, various enzymatic, immunological, and inflammatory processes abnormally degrade hyaluronic acid (HA), SF's main component, and affect the concentrations of specific proteins, with the final results seriously endangering synovial fluid (SF)'s rheological and tribological features and characteristics. No effective treatments have been found to stop the progression of KOA, but the injection of HA-based viscoelastic gels has been considered (alone or combined with physiotherapy (PT)) as an alternative to symptomatic therapies. In order to evaluate the effect of viscosupplementation and PT on the characteristics of SF, SF aspirated from groups treated for KOA (HA Kombihylan ® and groups that received Kombihylan ® and complex PT) was analyzed and compared from analytical, spectrophotometrical, and rheological perspectives. In the patients treated with PT, the SF extracted 6 weeks after viscosupplementation had a superior elastic modulus (G') and viscous moduli (G″), as well as a homogeneous distribution of proteins and polysaccharides. The viscosupplementation fluid improved the bioadhesive properties of the SF, and the use of the viscosupplementation fluid in conjunction with PT was found to be favorable for the distribution of macromolecules and phospholipids, contributing to the lubrication process and the treatment of OA-affected joints.

The importance of Saccharomyces cerevisiae yeast cells is known worldwide, as they are the most used microorganisms in biotechnology for bioethanol and biofuel production. Also, they are analyzed and studied for their similar internal biochemical processes to human cells, for a better understanding of cell aging and response to cell stressors. The special ability of S. cerevisiae cells to develop in both aerobic and anaerobic conditions makes this microorganism a viable model to study the transformations and the way in which cellular metabolism is directed to face the stress conditions due to environmental changes. Thus, this review will emphasize the effects of oxidative, ethanol, and osmotic stress and also the physiological and genetic response of stress mitigation in yeast cells.

Folic acid (vitamin B9) is an essential micronutrient for human health. It can be obtained using different biological pathways as a competitive option for chemical synthesis, but the price of its separation is the key obstacle preventing the implementation of biological methods on a broad scale. Published studies have confirmed that ionic liquids can be used to separate organic compounds. In this article, we investigated folic acid separation by analyzing 5 ionic liquids (CYPHOS IL103, CYPHOS IL104, [HMIM][PF 6 ], [BMIM][PF 6 ], [OMIM][PF 6 ]) and 3 organic solvents (heptane, chloroform, and octanol) as the extraction medium. The best obtained results indicated that ionic liquids are potentially valuable for the recovery of vitamin B9 from diluted aqueous solutions as fermentation broths; the efficiency of the process reached 99.56% for 120 g/L CYPHOS IL103 dissolved in heptane and pH 4 of the aqueous folic acid solution. Artificial Neural Networks (ANNs) were combined with Grey Wolf Optimizer (GWO) for modelling the process, considering its characteristics.

Experimental studies on the reactive extraction of fumaric acid with Amberlite LA-2 from Rhizopus oryzae suspensions using three solvents with different dielectric constants varying from 9.08 to 1.90 (dichloromethane, n-butyl acetate, and n-heptane, respectively) underlined the particular behavior of the extraction system in the presence of fungal biomass. The interfacial mass flow of the reaction product was found to be significantly affected by the biomass, due mainly to its adsorption onto the phase separation interface, this leading to the appearance of a physical barrier against the solute's transfer. However, the magnitude of the adsorption phenomenon was found to depend on Rhizopus oryzae 's affinity for the solvent phase, which increased significantly from dichloromethane to n-heptane. The negative influence of the biomass on the interfacial mass transfer can be partially counteracted by adding 1-octanol into the organic phase, improving the solvent's ability to solve the fumaric acid-Amberlite LA-2 complex and simplifying the reactive extraction mechanism, effects that were found to be more important for low-polar solvents. Consequently, for the same mixing intensity, the maximum amplification factor was reached for n-heptane, its value being almost 5-6 times higher than that obtained for dichloromethane and over 2 times higher than that obtained with n-butyl acetate.

Combining artificial neural networks with evolutive/bioinspired approaches is a technique that can solve a variety of issues regarding the topology determination and training for neural networks or for process optimization. In this chapter, the main mechanisms used in neuroevolution are discussed and some biochemical separation examples are given to underline the efficiency of these tools. For the current case studies (reactive extraction of folic acid and pertraction of vitamin C), the bioinspired metaheuristic included in the neuroevolutive procedures is represented by Differential Evolution, an algorithm that has shown a great potential to solve a variety of problems from multiple domains.

The addition of n -dodecane (between 1-3%) to the Escherichia coli fermentation broth in a mechanically agitated and aerated bioreactor revealed improved DO (dissolved oxygen) levels induced during fermentation which lead to an increase in biomass productivity and faster glucose consumption. The maximum values for enzyme activity (increased with 43% compared with the control) and k L a (the volumetric mass transfer coefficient) were obtained for the addition of 2% v/v n-dodecane in the bioreactor, due to the fact that oxygen limitation during the exponential growth phase of the bacterium can repress β-galactosidase production. The oxygen vector addition increased the available dissolved oxygen and activated a redox-sensitive regulation and an elevated intracellular oxidative metabolism that lead to the enhancement in E. coli biomass accumulation and a more accurate protein folding of β-galactosidase that would increase its activity. In addition to the experimental analysis, a complex model, developed using an improved version of Bacterial Foraging Algorithm and Artificial Neural Networks, was proposed, with a good average absolute value (6.2% in the training phase and 7.28% in the testing phase) between the process dynamic and the predictions generated by the model., Competing Interests: Conflict of interestThe authors declare that they have no conflict of interest in the publication., (© King Abdulaziz City for Science and Technology 2020.)

Mupirocin is an antibiotic from monocarboxylic acid class used as antibacterial agent against methicillin-resistant Staphylococcus aureus (MRSA) and can be obtained as a mixture of four pseudomonic acids by Pseudomonas fluorescens biosynthesis. Nowadays improving antibiotics occupies an important place in the pharmaceutical industry as more and more resistant microorganisms are developing. Mupirocin is used to control the MRSA outbreaks, for infections of soft tissue or skin and for nasal decolonization. Due to its wide use without prescription, the microorganism's resistance to Mupirocin increased from up to 81%, thus becoming imperative its control or improvement. As the biotechnological production of Mupirocin has not been previously reviewed, in the present paper we summarize some consideration on the biochemical process for the production of pseudomonic acids (submerged fermentation and product recovery). Different strains of Pseudomonas, different culture medium and different conditions for the fermentation were analysed related to the antibiotics yield and the product recovery step is analysed in relation to the final purity. However, many challenges have to be overcome in order to obtain pseudomonic acid new versions with better properties related to antibacterial activity.

Erythromycin has been separated from simulated broths by free and facilitated pertraction with D2EHPA dissolved in dichloromethane using an U-shaped pertraction cell. The pertraction has been analyzed by means of initial and final mass flows and permeability factor. The main factors which control the pertraction efficiency were identified to be the pH-gradient between the feed and stripping phase, aqueous phases viscosities and mixing intensity.

The study on mixing distribution for an aerobic stirred bioreactor and simulated (solutions of carboxymethylcellulose sodium salt), yeasts (S. cerevisiae) and fungus (P. chrysogenum pellets and free mycelia) broths indicated the significant variation of mixing time on the bioreactor height. The experiments suggested the possibility to reach a uniform mixing in whole bulk of the real broths for a certain value of rotation speed or biomass concentration domain. For S. cerevisiae broths the optimum rotation speed increased to 500 rpm with the biomass accumulation from 40 to 150 g/l d.w. Irrespective of their morphology, for fungus cultures the existence of optimum rotation speed (500 rpm) has been recorded only for biomass concentration below 24 g/l d.w. The influence of aeration rate depends on the apparent viscosity/biomass concentration and on the impellers and sparger positions. By increasing the apparent viscosity for simulated broths, or biomass amount for real broths, the shape of the curves describing the mixing time variation is significantly changed for all the considered positions. The intensification of the aeration induced the increase of mixing time, which reached a maximum value, decreasing then, due to the flooding phenomena. This variation became more pronounced at higher viscosities for simulated broths, at higher yeasts concentration, and at lower pellets or filamentous fungus concentration, respectively. By means of the experimental data and using MATLAB software, some mathematical correlations for mixing time have been proposed for each broth and considered position inside the bioreactor. These equations offer a good agreement with the experiment, the maximum deviation being +/-7.3% for S. cerevisiae broths.

The previous works on simulated broths are continued and developed for Propionibacterium shermanii broths. The obtained results indicated the considerable increase of kLa in presence of n-dodecane as oxygen-vector and the existence of a certain value of hydrocarbon concentration that corresponds to the maximum mass transfer rate of oxygen. The magnitude of the positive effect of the oxygen-vector strongly depends on operational conditions of the bioreactor, on broth characteristics and on P. shermanii concentration.

Oxygen mass transfer represents the most important parameter involved in the design and operation of mixing-sparging equipment for bioreactors. It can be described and analyzed by means of the mass transfer coefficient, k(L) a. The k(L) a values are affected by many factors such as geometrical and operational characteristics of the vessels, media composition, type, concentration and microorganism morphology, and biocatalysts properties. The efficiency of oxygen transfer could be enhanced by adding oxygen-vectors in broths, such as hydrocarbons or fluorocarbons, without increasing the energy consumption for mixing or aeration. The experimental results obtained for simulated broths indicated a considerable increase of k(L) a in the presence of n-dodecane, and the existence of a certain value of n-dodecane concentration that corresponds to a maximum mass transfer rate of oxygen. The magnitude of the positive effect of n-dodecane depends both on the broths' characteristics and operational conditions of the bioreactor.