Standing wave design and experimental validation of a tandem simulated moving bed process for insulin purification

A tandem simulated moving bed (SMB) process for insulin purification has been proposed and validated experimentally. The mixture to be separated consists of insulin, high molecular weight proteins, and zinc chloride. A systematic approach based on the standing wave design, rate model simulations, and experiments was used to develop this multicomponent separation process. The standing wave design was applied to specify the SMB operating conditions of a lab-scale unit with 10 columns. The design was validated with rate model simulations prior to experiments. The experimental results show 99.9% purity and 99% yield, which closely agree with the model predictions and the standing wave design targets. The agreement proves that the standing wave design can ensure high purity and high yield for the tandem SMB process. Compared to a conventional batch SEC process, the tandem SMB has 10% higher yield, 400% higher throughput, and 72% lower eluant consumption. In contrast, a design that ignores the effects of mass transfer and nonideal flow cannot meet the purity requirement and gives less than 96% yield.     

Separation of amino acids by simulated moving bed using competitive Langmuir isotherm

The separation of two amino acids, phenylalanine and tryptophan, was carried out using laboratory simulated moving bed (SMB) chromatography. The SMB process consisted of four zones, with each zone having 2 columns. The triangle theory was used to obtain the operating conditions for the SMB. The mass transfer coefficients of the two amino acids were obtained from the best-fit values by comparing simulated and experimental pulse data. The competitive adsorption isotherms of the two amino acids were obtained by single and binary frontal analyses, taking into consideration the competition between the two components. A competitive Langmuir isotherm, obtained from single-component frontal chromatography, was used in the first run, and the isotherm from binary frontal chromatography in the second, with the flow rate of zone I modified to improve the purity. Compared to the first and second runs, the competitive Langmuir isotherm from the binary frontal chromatography showed good agreement with the experimental results. Also, adjusting the flow rate in zone I increased the purity of the products. The purities of the phenylalanine in the raffinate and the tryptophan in the extract were 99.84 and 99.99%, respectively.     

Continuous separation of succinic acid and lactic acid by using a three-zone simulated moving bed process packed with Amberchrom-CG300C

The issue of separating succinic acid and lactic acid in a continuous mode has been a major concern in the biotechnological process for production of succinic acid. To address this issue, both the optimal design and the experimental validation of a three-zone simulated moving bed (SMB) process for such separation were attempted in this article using the Amberchrom-CG300C resin and a self-assembled SMB unit with three zones. First, the intrinsic parameters of the two organic acids on the Amberchrom-CG300 resin were estimated at 40 °C from a series of multiple frontal experiments. The resulting intrinsic parameters were then used in optimizing the experimental setting points for pump flow rates and switching time of the three-zone SMB equipment, which was assisted by an up-to-date genetic algorithm. Based on the optimized conditions, the relevant SMB experiment was conducted at 40 °C and all the resultant samples from the product ports and column outlets of the SMB unit were assayed. It was confirmed from the assay results that the continuous separation of succinic acid and lactic acid was performed successfully. The experimental data for the product concentration profiles and the internal concentration profiles were also in reasonable agreement with the model predictions.     

Simulation and operation performance of simulated moving bed for enatioseparation of mandelic acid

Separation of mandelic acid enantiomer was carried out by using laboratory simulated moving bed (SMB) chromatography. The SMB process consists of four zones, with each zone having 1 column. The triangle theory was used to obtain the operating conditions for SMB. Adsorption isotherms of the D-/L-mandelic acids were obtained by pulse input method, considering linear isotherms of the two components. Flow rates of extract, raffinate, feed, and eluent streams were systematically changed to understand the effects of operation flow rates of SMB. Simulation results and experimental data from the SMB chromatography showed good agreements. Adjusting the flow rate in zone II increased the purity of D-mandelic acid from extract port. The highest purity of D-mandelic acid in the extract was obtained as 94% under the operating flow rates of Q_feed = 0.1 mL/min, Q_extract = 0.2 mL/min, Q_raffinate = 0.8 mL/min, and Q_eluent = 0.9 mL/min.     

Microbial production of d‐lactic acid from dried distiller's grains with solubles

d ‐Lactic acid production is gaining increasing attention due to the thermostable properties of its polymer, poly‐d ‐lactic acid . In this study, Lactobacillus coryniformis subsp. torquens, was evaluated for its ability to produce d ‐lactic acid using Dried Distiller's Grains with Solubles (DDGS) hydrolysate as the substrate. DDGS was first subjected to alkaline pretreatment with sodium hydroxide to remove the hemicellulose component and the generated carbohydrate‐rich solids were then subjected to enzymatic hydrolysis using cellulase mixture Accellerase® 1500. When comparing separate hydrolysis and fermentation and simultaneous saccharification and fermentation (SSF) of L. coryniformis on DDGS hydrolysate, the latter method demonstrated higher d ‐lactic acid production (27.9 g/L, 99.9% optical purity of d ‐lactic acid), with a higher glucose to d ‐lactic acid conversion yield (84.5%) compared to the former one (24.1 g/L, 99.9% optical purity of d ‐lactic acid). In addition, the effect of increasing the DDGS concentration in the fermentation system was investigated via a fed‐batch SSF approach, where it was shown that the d ‐lactic acid production increased to 38.1 g/L and the conversion yield decreased to 70%. In conclusion, the SSF approach proved to be an efficient strategy for the production of d ‐lactic acid from DDGS as it reduced the overall processing time and yielded high d ‐lactic acid concentrations.     

Optimal design and experimental validation of a modified four-zone SMB process for the separation of a ternary amino acid mixture

For a standard four-zone simulated moving bed (SMB) chromatography, several process modifications have been made in previous studies such that its application scope could be extended to a ternary separation. One of the effective modifications reported was to (1) replace its closed-loop configuration with an open-loop configuration and (2) utilize the extract port for collecting both the intermediate-affinity and the highest-affinity components in regular sequence in every switching period. Most of previous researches on such a modified four-zone SMB (MF-SMB) have been limited to process simulation and optimization. The experimental validation of the MF-SMB process with linear isotherms was attempted in this article for the first time using a ternary amino acid mixture as a model system. First, the intrinsic parameters of three amino acids were estimated from a series of multiple-frontal experiments. The estimated parameter values were then used in the stage of the MF-SMB optimization, which was assisted by an up-to-date genetic algorithm. Based on the optimized conditions, the MF-SMB experiment was conducted and the assay results for product samples verified the attainment of a ternary separation. The experimental purities and concentrations were also found to agree closely with the model predictions.     

Efficient homofermentative L-(+)-lactic acid production from xylose by a novel lactic acid bacterium, Enterococcus mundtii QU 25

Enterococcus mundtii QU 25, a newly isolated lactic acid bacterium, efficiently metabolized xylose into l-lactate. In batch fermentations, the strain produced 964 mM l-(+)-lactate from 691 mM xylose, with a yield of 1.41 mol/mol xylose consumed and an extremely high optical purity of ≥99.9% without acetate production.     

Green biorefinery: separation of lactic acid from grass silage juice by chromatography using neutral polymeric resin

The aim of this work was to recover lactic acid in undissociated form from grass silage juice. For this aim, chromatographic separation using neutral polymeric resin Amberlite XAD1600 was investigated. Up to now, there is no hint in the literatures about using neutral polymeric resin for lactic acid separation from a mixture. Important factors (flow-rate, concentration of feed and loaded volume) that affect separation performance were firstly investigated with model solutions. The obtained results showed that lactic acid solutions with the purity varying from 93.2% to 99.9% could be obtained at the recovery yields over 99.4%. After that, trials with silage juice were carried out. Due to the complex composition of the feed, the purity of products decreased to 94% at a recovery yield of 97%. Although 99% of inorganic salts and sugars were separated from lactic acid organic acids in general and acetic acid in particular caused a purity problem. It seems that organic acids could not be separated from lactic acid by neutral resin Amberlite XAD1600. Besides the organic acid problem, some amino acids were remained in the products as impurities.     

Development of a three-zone simulated moving bed process based on partial-discard strategy for continuous separation of valine from isoleucine with high purity,high yield,and high product concentration

The issue of separating valine from isoleucine has been a major concern in the biotechnological process for production of valine. To address this issue, an optimal three-zone simulated moving bed (SMB) process for continuous separation of valine was developed in this study. It was first found that an Amberchrom-CG161C resin was highly suitable for the adsorbent of such SMB process. The adsorption isotherm and mass-transfer parameters of valine and isoleucine on the Amberchrom-CG161C adsorbent were then determined through multiple frontal experiments. The determined parameters were used in the next stage of optimizing the SMB for valine separation, which was performed on the basis of genetic algorithm. For the optimized SMB process, a partial-discard strategy was applied to the raffinate port in order to make a further improvement in the valine product concentration. Finally, the optimized SMB based on the partial-discard strategy was tested experimentally using the self-assembled SMB equipment. The experimental results showed that the developed process in this study was highly effective in continuous separation of valine from isoleucine while ensuring the attainment of high product concentration. The experimental data for the SMB effluent histories and the SMB column profiles were also in close agreement with the model predictions.     

pH-Uncontrolled lactic acid fermentation with activated carbon as an adsorbent

In this paper, we presented a novel process involving activated carbon (AC) as an adsorbent for lactic acid fermentation, separation and oligomerization. It was found that pH has a significant effect on the adsorption of lactic acid on AC. The use of AC for in situ removal of lactic acid successfully decreased the inhibitory effect of lactic acid, resulting in significant increases in both productivity and yield. Acetone was used to desorb lactic acid and it was confirmed that the acetone treatment did not decrease the optical purity of the lactic acid, i.e., the optical purity was as high as 99.5% after desorption. Due to the presence of little materials influencing lactic acid oligomerization, oligomers with an optical purity of above 96% and a weight-average molecular weight (M(w)) of 2400 were obtained in the oligomerization process.     

Optimal design and experimental validation of a three-zone simulated moving bed process based on the Amberchrom-CG161C adsorbent for continuous removal of acetic acid from biomass hydrolyzate

In the production process of bio-ethanol from biomass, acetic acid is recognized as the key impurity to be removed from the sugar components that are generated by hydrolyzing biomass. In regard to this issue, it has recently been confirmed that the Amberchrom-CG161C resin was highly qualified as the adsorbent of a simulated moving bed (SMB) process for continuous separation of acetic acid from the biomass hydrolyzate, i.e., sugars. However, the previous study on the Amberchrom-CG161C SMB with the aforementioned separation goal has been limited to only a theoretical work, including some batch-chromatography tests. The experimental validation of such an Amberchrom-CG161C SMB process, including its optimal design, was attempted in this article. This task began by assembling the experimental unit of the SMB process with three zones. Its operating conditions were then optimized by using genetic algorithm. Under the optimized operating conditions, the relevant three-zone SMB experiment was conducted. The assay of all the resultant product samples verified that the SMB separation of interest was performed successfully as designed. The experimental data were also found to agree closely with the model predictions. Finally, a partial-discard strategy was applied to maintain the sugar product concentration as high as possible.     

Sensitivity analysis of amino acids in simulated moving bed chromatography

We conducted a sensitivity analysis of the simulated moving bed (SMB) chromatography with the case model of the separation of two amino acids phenylalanine and tryptophan. We consider a four-zone SMB chromatography where the triangle theory is used to determine the operating conditions. Competitive Langmuir isotherm model was used to determine the adsorption isotherm. The finite difference method is used to solve nonlinear partial differential equation (PDE) systems numerically. We examined the effects of alterations in the operating conditions (feed-extract, feed-raffinate, eluent-extract, eluent-raffinate, recycle, and switching time) and the adsorption isotherm parameters (Langmuir isotherm parametersa andb) on SMB efficiency. The variation range of operating conditions and Langmuir isotherma was between −50 and 50% of original value and the variation range of the Langmuir isothermb was between 2.25⁻⁵ and 2.25⁵ times of original value.     

Prediction of band profiles of mixtures of bradykinin and kallidin from data acquired by competitive frontal analysis

The competitive adsorption isotherms of two closely related peptides, bradykinin and kallidin, were measured by frontal analysis on a Zorbax SB-C18 microbore column. An aqueous soluton at 20% acetonitrile (0.1% TFA) was used as the mobile phase. The competitive isotherm data were fitted to four different models: Langmuir, Bilangmuir, Langmuir-Freundlich, and Toth. These data fitted best to a Bilangmuir isotherm model. The influence of the pressure on the retention factors of the two peptides was found to be small and was not investigated in detail. The band profiles of large samples of the single components and of their mixtures were recorded. The overloaded profiles calculated using either the equilibrium-dispersive or POR model are in excellent agreement with the experimental profiles in all cases. Our results confirm that the competitive isotherm data derived from mixtures may suffice for a reasonably accurate prediction of the band profiles of all mixtures of the two components, provided their composition is close to 1/1.     

Equilibrium and a two-stage batch adsorber design for reactive or disperse dye removal to minimize adsorbent amount

The adsorption of a reactive dye (Reactive Yellow K-4G) and a disperse dye (Disperse yellow brown S-2RFL) onto polyepicholorohydrin-dimethylamine (EPIDMA) cationic polymer modified bentonite (EPIDMA-bentonite) in batch adsorber was studied, respectively. Two equilibrium models, the Langmuir and Freundlich models were selected to follow the adsorption process. It was shown that the equilibrium experimental data for reactive dye adsorption could be well described by the Freundlich model, but for disperse dye the Langmuir model could be better. Based on the well correlated adsorption isotherm, an adsorption process design model was developed for the design of a two-stage batch adsorber to predict the minimum amount of adsorbent to achieve a specified percentage of dye removal at a given volume of wastewater effluents. The adsorption process design analysis indicated that compared with the single-stage batch adsorption, the two-stage process could significantly save adsorbent to meet the higher demands of dye removal efficiency.     

Genetically engineered wine yeast produces a high concentration of L-lactic acid of extremely high optical purity

For mass production of lactic acid, we newly constructed a transgenic wine yeast strain that included six copies of the bovine L-lactate dehydrogenase gene on the genome. On fermentation in inexpensive cane juice-based medium, L-lactate production of this recombinant reached 122 g/liter and the optical purity was 99.9% or higher.     

Modified reactive SMB for production of high concentrated fructose syrup by isomerization of glucose to fructose

This work presents modifications to the Hashimoto's hybrid simulated moving bed reactor (SMBR) system which was used to produce 55% high fructose syrup (HFS55). The purpose of this study is to develop a new SMBR system to overcome the disadvantages of Hashimoto system (3-zone SMB with seven reactors), i.e., low utility of reactors when feed being a 50/50 blend of glucose and fructose. Two different configurations of modified system were presented in this paper: the first configuration is 4-zone SMB with one reactor, while the other one consists of one additional reactor. Both of these configurations aim at improving the concentration and purity of glucose at the inlet of the reactor, which will lead to both high productivity and high purity of fructose in the product. A state-of-the-art optimization technique, viz., non-dominated sorting genetic algorithm (NSGA) is used in finding the optimal design and operating parameters for the modified reactive SMB and Varicol processes. Compared with the Hashimoto's system, high productivity and purity of fructose can be achieved in these new systems using less number of reactors.     

Production of <Emphasis Type="SmallCaps">l</Emphasis>-lactic acid from a mixture of xylose and glucose by co-cultivation of lactic acid bacteria

The production of optically pure lactic acid in a high yield from xylose or a mixture of xylose and glucose, which is a model hydrolysate of lignocellulose, is described. In a single cultivation, Enterococcus casseliflavus produced 38 g/l of lactic acid with an optical purity of 96% enantiomeric excess (ee) and 6.4 g/l of acetic acid from 50 g/l of xylose when MRS medium was used. When a mixture of 50 g/l of xylose and 100 g/l of glucose was used as the carbon source in a cultivation of E. casseliflavus alone, glucose was converted to lactic acid in the early phase of the cultivation but xylose was hardly consumed. In a co-cultivation where E. casseliflavus and Lactobacillus casei specific for glucose were simultaneously inoculated, little or no lactic acid was produced after the glucose was almost consumed. A co-cultivation with two-stage inoculation (in which E. casseliflavus was added at a cultivation time of 40 h after L. casei cells were inoculated) resulted in complete consumption of 50 g/l of xylose and 100 g/l of glucose. In the co-cultivation, 95 g/l of lactic acid with a high optical purity of 96% ee was obtained at 192 h. Such a co-cultivation using two microorganisms specific for each sugar is considered to be one promising cultivation technique for the efficient production of lactic acid from a sugar mixture derived from lignocellulose.     

米根霉发酵生产L-乳酸

报道了Ｌ－乳酸菌株的分离与筛选,探讨了不同碳源、氮源、通气量、温度等发酵条件对产Ｌ－乳酸的影响,从７８株米根霉中筛选出１３株产Ｌ－乳酸较高的菌株,其中米根霉（Ｒｈｉｚｏｐｕｓ ｏｒｙｚａｅ）Ｒｓ９２８产Ｌ－乳酸最高,产酸最稳定。试验结果表明,该菌株最适发酵培养组成（％）：淀粉水解糖１６,ＭｇＳＯ４ ０．０８,ＫＨ２ＰＯ４ ０．０５,ＺｎＳＯ４ ０．０１,ＣａＣＯ３ ７,ｐＨ自然。在６０ｔ发酵罐中,     

Improvement of lactic acid production from cellulose with the addition of Zn/Ni/C under alkaline hydrothermal conditions

The effect of Zn, Ni and activated carbon on the yield of lactic acid from cellulose was investigated to improve the lactic acid yield under alkaline hydrothermal conditions. The results showed that the lactic acid yield increased greatly in the presence of Zn, Ni and activated carbon. Central composite response surface method (RSM) design experimentation was used to find the optimal concentrations of Zn, Ni, activated carbon and NaOH, which indicated that 0.02 g Zn, 0.03 g Ni, 0.07 g activated carbon and 2.5 mol/L NaOH were the optimal concentrations. Under this condition, the highest lactic acid yield was 42%, which was much higher than previous results using only NaOH (15%). The confirmatory experiments on lactic acid yield proved that the proposed model of lactic acid yield can accurately predict the lactic acid yield from cellulose.     

Isolation and characterisation of lactic acid bacterium for effective fermentation of cellobiose into optically pure homo <Emphasis Type="SmallCaps">l</Emphasis>-(+)-lactic acid

Effective utilisation of cellulosic biomasses for economical lactic acid production requires a microorganism with potential ability to utilise efficiently its major components, glucose and cellobiose. Amongst 631 strains isolated from different environmental samples, strain QU 25 produced high yields of l-(+)-lactic acid of high optical purity from cellobiose. The QU 25 strain was identified as Enterococcus mundtii based on its sugar fermentation pattern and 16S rDNA sequence. The production of lactate by fermentation was optimised for the E. mundtii QU25 strain. The optimal pH and temperature for batch culturing were found to be 7.0°C and 43°C, respectively. E. mundtii QU 25 was able to metabolise a mixture of glucose and cellobiose simultaneously without apparent carbon catabolite repression. Moreover, under the optimised culture conditions, production of optically pure l-lactic acid (99.9%) increased with increasing cellobiose concentrations. This indicates that E. mundtii QU 25 is a potential candidate for effective lactic acid production from cellulosic hydrolysate materials.