Extractive fermentation of acetic acid

In this technoeconomic evaluation of the manufacture of acetic acid by fermentation, the use of the bacterium:Acetobacter suboxydans from the old vinegar process was compared with expected performance of the newerClostridium thermoaceticum bacterium. Both systems were projected to operate as immobilized cells in a continuous, fluidized bed bioreactor, using solvent extraction to recover the product. Acetobacter metabolizes ethanol aerobically to produce acid at 100 g/L in a low pH medium. This ensures that the product is in the form of a concentrated extractable free acid, rather than as an unextractable salt. Unfortunately, yields from glucose by way of the ethanol fermentation are poor, but near the biological limits of the organisms involved.     

Simultaneous saccharification and extractive fermentation of lignocellulosic materials into lactic acid in a two-zone fermentor-extractor system

Simultaneous saccharification and extractive fermentation of lignocellulosic materials into lactic acid was investigated using a two-zone bioreactor. The system is composed of an immobilized cell reactor, a separate column reactor containing the lignocellulosic substrate and a hollow-fiber membrane. It is operated by recirculating the cell free enzyme (cellulase) solution from the immobilized cell reactor to the column reactor through the membrane. The enzyme and microbial reactions thus occur at separate locations, yet simultaneously. This design provides flexibility in reactor operation as it allows easy separation of the solid substrate from the microorganism, in situ removal of the product and, if desired, different temperatures in the two reactor sections. This reactor system was tested using pretreated switchgrass as the substrate. It was operated under a fed-batch mode with continuous removal of lactic acid by solvent extraction. The overall lactic acid yield obtainable from this bioreactor system is 77% of the theoretical.     

An electrokinetic bioreactor: using direct electric current for enhanced lactic acid fermentation and product recovery

The microbiological production of organic acids by fermentation processes is growing in commercial importance. However, the removal of product and pH control are two main issues that limit the technical and commercial viability of such processes. A laboratory scale bioreactor combining conventional electrodialysis and bipolar membrane electrodialysis has been developed for in situ product removal and pH control in lactic acid fermentation. The electrokinetic process enabled removal of the biocatalytic product (lactic acid) directly from the bioreactor system, in a concentrated form, as well as enabling good pH control without generation of troublesome salts. Moreover, end-product inhibition of glucose catabolism was reduced, resulting in a greater generation of the end-product lactic acid. An automatic pH sensor and current application system was developed and successfully implemented for lactic acid fermentation in the electrokinetic bioreactor.     

Lactic acid production from cheese whey by immobilized bacteria

The performance of immobilized Bifidobacterium longum in sodium alginate beads and on a spiral-sheet bioreactor for the production of lactic acid from cheese whey was evaluated. Lactose utilization and lactic acid yield of B. longum were compared with those of Lactobacillus helveticus. B. longum immobilized in sodium alginate beads showed better performance in lactose utilization and lactic acid yield than L. helveticus. In the spiral-sheet bioreactor, a lactose conversion ratio of 79% and lactic acid yield of 0.84 g of lactic acid/g of lactose utilized were obtained during the first run with the immobilized L. helveticus. A lactose conversion ratio of 69% and lactic acid yield of 0.51 g of lactic acid/g of lactose utilized were obtained during the first run with immobilized B. longum in the spiral-sheet bioreactor. In producing lactic acid L. helveticus performed better when using the Spiral Sheet Bioreactor and B. longum showed better performance with gel bead immobilization. Because B. longum is a very promising new bacterium for lactic acid production from cheese whey, its optimum fermentation conditions such as pH and metabolic pathway need to be studied further. The ultrafiltration tests have shown that 94% of the cell and cheese whey proteins were retained by membranes with a mol wt cutoff of 5 and 20 KDa.     

Repeated Batch Cell-Immobilized System for the Biotechnological Production of Xylitol as a Renewable Green Sweetener

The present paper studies the biotechnological production of xylitol using sugarcane bagasse hydrolysate in a repeated batch fermentation system with immobilized cells of Candida guilliermondii FTI20037. Immobilized cell system is considered as an attractive alternative to reuse the well-grown and adapted yeast cells in a new fresh fermentation media, without the need of the inoculum stage. In this work, seven repeated batches were performed in a fluidized bed bioreactor using immobilized cells in calcium alginate beads. According to the obtained results it was observed that the immobilized cells of C. guilliermondii can be reused for six successive batches maintaining an average xylitol yield (Y _p/s) of 0.7 g/L and a volumetric productivity (Q _p) of 0.42 g/L?h at the end of 432 h of fermentation. On the other hand, in the seventh batch (504 h), a decrease of 44 % in the final concentration of xylitol was observed. This reduction can be explained by the possible diffusion and accumulation of insoluble substances, found in the hemicellulosic hydrolysate, in the interior of the immobilization support resulting in substrate mass transfer limitations.     

用于流化床燃烧脱硫的石灰石的反应活性评价和测试研究

对用于燃煤流化床燃烧脱硫的脱硫剂石灰石的反应活性进行了研究，提出了易于进行数学处理的石灰石硫盐化模型，得出了评价石灰石反应活性的两个指标－最大转化率和反应速率常数。研究方法除采用了传统的热天平法和鼓泡流化床外，还根据循环流化床燃烧技术的特点，发展了提出了湍流床法和石英棉法，并进行了不同试验条件下的实验研究，对不同反应活性温度技术进行了试验比较。     

Lactic acid fermentation in cell-recycle membrane bioreactor

Traditional lactic acid fermentation suffers from low productivity and low product purity. Cell-recycle fermentation has become one of the methods to obtain high cell density, which results in higher productivity. Lactic acid fermentation was investigated in a cell-recycle membrane bioreactor at higher substrate concentrations of 100 and 120 g/dm³. A maximum cell density of 145 g/dm³ and a maximum productivity of 34 g/(dm³…h) were achieved in cell-recycle fermentation. In spite of complete consumption of substrate, there was a continuous increase in cell density in cell-recycle fermentation. Control of cell density in cell-recycle fermentation was attempted by cell bleeding and reduction in yeast extract concentration.     

Production of Butyric Acid from Glucose and Xylose with Immobilized Cells of <Emphasis Type="Italic">Clostridium tyrobutyricum</Emphasis> in a Fibrous-bed Bioreactor

Butyric acid has many applications in chemical, food, and pharmaceutical industries. In the present study, Clostridium tyrobutyricum ATCC 25755 was immobilized in a fibrous-bed bioreactor to evaluate the performance of butyrate production from glucose and xylose. The results showed that the final concentration and yield of butyric acid were 13.70 and 0.46 g g⁻¹, respectively, in batch fermentation when 30 g L⁻¹ glucose was introduced into the bioreactor. Furthermore, high concentration 10.10 g L⁻¹ and yield 0.40 g g⁻¹ of butyric acid were obtained with 25 g L⁻¹ xylose as the carbon source. The immobilized cells of C. tyrobutyricum ensured similar productivity and yield from repeated batch fermentation. In the fed-batch fermentation, the final concentration of butyric acid was further improved to 24.88 g L⁻¹ with one suitable glucose feeding in the fibrous-bed bioreactor. C. tyrobutyricum immobilized in the fibrous-bed bioreactor would provide an economically viable fermentation process to convert the reducing sugars derived from plant biomass into the final bulk chemical (butyric acid).     

Lactic Acid Production from a Whole Slurry of Acid-Pretreated Spent Coffee Grounds by Engineered Saccharomyces cerevisiae

Spent coffee grounds (SCG) generated after coffee extraction are the main byproduct of the coffee industry. Valorization of the SCG has been increasingly focused following considerable attention in coffee consumption. Lactic acid bacteria fermentation is the primary source of generation of lactic acid, a monomer of polylactic acid that has various industrial applications; however, because of the low tolerance of lactic acid bacteria to toxic compounds, it is necessary to apply Saccharomyces cerevisiae to produce lactic acid whose tolerance to toxic compounds is higher. In this study, we evaluated the feasibility of using SCG as substrate for the production of lactic acid by S. cerevisiae strain expressing heterologous lactate dehydrogenase. The fermentation profiles of the engineered yeast showed that lactic acid production was promoted by xylose addition. From simultaneous saccharification and fermentation (SSF) using a whole slurry of acid-pretreated SCG, containing high amounts of hemicellulose fractions, lactic acid (0.11 g) and ethanol (0.10 g) per g SCG were obtained after 24 h of SSF, of which yields were 413% and 221% higher, respectively, than those of washed pretreated SCG. Thus, fermentation of whole slurry SCG by engineered S. cerevisiae is a suitable way of lactic acid production, selectively.

     

Technoeconomics of butanol extractive fermentation in a multiphase fluidized bed bioreactor

The design, performance, and projected economics of a new conceptual process for thein situ solvent extraction of butanol within a multiphase fluidized bed bioreactor are discussed. The process appears to have the potential to increase the effective concentration of product over tenfold while maintaining actual concentrations in contact with the organism at 10 g/L, i.e., below the threshold of inhibition. If this performance can be realized in commercial operation, the selling price to yield a 30% pretax return on investment could be reduced from over $1.25 per pound of butanol for the present process to $0.59 per pound for the new process.

     

Strategies in lipase production by immobilizedCandida rugosa cells

Growing cells ofCandida rugosa immobilized in polymethacrylamide-hydrazide and polyurethane foam were employed in fluidized and packed bed reactors, for discontinuous and continuous fermentations to obtain extracellular lipase. In spite of hydrodynamic problems, fermentation cultures using polyurethane foam showed higher lipolytic activity than cultures employing polymethacrylamide-hydrazide beads, which was probably owing to the high immobilized biomass concentration in polyurethane observed by direct microscopy enumeration. Different oleic acid concentrations were assayed. The maximum level of lipase was achieved at 4 g/L of oleic acid. These results reaffirm that lipase production is a direct function of cell-substrate contact and that the organic substrate dispersion is important in this system.     

Hydrodynamics and mixing in three-phase fluidized bed bioreactors

The effects of solid particles on hydrodynamics and mixing in a three-phase fluidized bed bioreactor were discussed. The gas holdup, bubble size, and liquid-phase axial dispersion coefficient were measured in a 0.25-m id bubble column bioreactors containing low-density particles. The presence of low-density solid particles slightly increased gas holdup. The decrease in average bubble diameter with solid presence was found. For the three-phase system, the liquid-phase axial dispersion coefficients were higher than for the two-phase system. We extended a model for a gas holdup developed for a gas-liquid two-phase bubble column bioreactor to a gas-liquid-solid three-phase fluidized bed bioreactor. Using the present data and available data in the literature, the predictions of the proposed model were examined. The proposed model agreed with a wide range of the experimental data. A theoretical correlation for liquid-phase axial dispersion coefficient was developed using Kolmogoroff's theory of isotropic turbulence. Reasonable agreement was obtained between the predicted and experimental values of axial dispersion coefficient.     

Particle circulation and oxygen mass transfer in an immobilized cell bioreactor

Two important considerations in the design of an aerobic particulate immobilized cell bioreactor are the provision of sufficient oxygen to maintain the desired metabolism of the immobilized organism, and the biomass holdup (which is proportional to the number of immobilized cell particles in the reactor). The Circulating Bed Reactor, a reactor developed for use with those forms of immobilization that result in particles of essentially neutral buoyancy, operates with an expanded bed of circulating particles. The particle number density attainable in such a reactor has been found to be dependent upon the circulation cell aspect ratio, the individual particle properties, the static bed voidage of the particles, and the superficial gas velocity. The oxygen mass transfer characteristics have been found to be dependent upon the circulatory nature of the system, the particle (solids) holdup, the particle porosity, and the superficial gas velocity.     

Production of Lactic Acid from Raw Sweet Potato Powders by Rhizopus Oryzae Immobilized in Sodium Alginate Capsules

Rhizopus oryzae immobilized in calcium alginate was applied in lactic acid fermentation with unhydrolyzed raw sweet potato powders as the sole carbon source. The effects of sodium alginate concentration, calcium chloride concentration, and the immobilized bead diameter on lactic acid production were investigated. Increase in sodium alginate concentration during the gelation process would harden the immobilized capsule, which led to a decrease in lactic acid production. The increase in calcium chloride would increase the thickness of the immobilized capsule, which would increase the mass transfer resistance. Nevertheless, while the calcium chloride was lower than 15%, it would not have obvious effects on lactic acid production. A larger bead could have more space for cell growth, which led to the maximum lactic acid production observed at the 5-mm bead diameter. Moreover, results of repeated-batch operation suggested that immobilized cells could have higher stability in lactic acid production than free cells. The total cumulative lactic acid in immobilized-cell operation could increase by 55% as compared with free-cell operation after 216 h (seven repeated-batches), and no loss of amylolytic activity was observed. The results indicated that immobilized R. oryzae by Ca-alginate could be suitable for lactic acid production from unhydrolyzed raw potato powders.     

An immobilized microbial heparinase for blood deheparinization

A new medical application of an immobilized microbial enzyme is described. Extracorporeal devices require systemic heparin administration to prevent thrombus formation; however, the use of heparin often leads to serious hemorrhagic complica tions. Heparinase isolated from Flavobacterium has been immobilized and used in a fluidized bed reactor to eliminate heparin from blood passing through an extracorporeal circuit both in vitro and in vivo. This paper discusses the stepwise de velopment of this heparinase reactor including: (1) improvements in the fermentation resulting in an inexpensive large-scale source of heparinase without the addition of the previously required inducer, heparin; (2) the use of batch processes to adapt previous purification schemes to large-scale heparinase production and the subsequent purifica tion of heparinase to a single SDS-PAGE banding protein; (3) the immobilization of heparinase with a 91% activity recovery and good stability, (4) the design and suc cessful testing of a fluidized bed reactor containing immobilized heparinase in the re moval of clinically used quantities of heparin from both human blood in vitro and ca nine blood in vivo; and (5) the initiation of animal studies focusing on the toxicology of heparinase-derived heparin degradation products and the short and long term effects of exposure to these products and to heparinase.     

Simulation of a hybrid fermentation-separation process for production of butyric acid

Simulation of a hybrid fermentation-separation process for the production of butyric acid (BA) based on published data was done. A unit consisting of a bioreactor with immobilized cells in the fibrous bed and of separation by pertraction through supported liquid membranes (SLM) was considered. Productivities of the unit volume of a fixed bed bioreactor in continuous and fed-batch fermentation at pH 5.5 and 6.0 were used. Concentration of BA in the bioreactor outlet stream was assumed to be in the interval from 0.11 kmol m⁻³ to 0.45 kmol m⁻³. Data on the pertraction through SLM with phosphonium ionic liquid (IL) and bulk liquid membrane with trioctylamine (TOA) as carriers were used. A strong increase in the required membrane area was found for both carriers at the pH of pertraction above 4. pH values of fermentation and pertraction should be optimized independently. It is advantageous to have pH of the feed into the pertraction unit of about 4. Dependences of the membrane area on the pertraction efficiency are nearly linear and not very sharp, especially for IL, what enables working at the pertractor efficiency exceeding 90 %. Application of phosphonium IL is promising compared to classical extractant TOA because of lower demand of the membrane area in a large interval of BA concentrations in the pertractor feed.     

Development of continuous fermentation using immobilized yeast cells for wine cooler process development

The continuous wine fermentation process, which employs a newly designed tapered column type bioreactor and immobilized yeast cells (Montrachet 522), was studied and its fermentation performance was compared with batch and suspended cell continuous wine fermentation systems. It was found that a stable continuous culture fermentation process could be maintained for a period of 2–3 mo when the new bioreactor system packed with immobilized yeast cells was employed. The new bioreactor containing immobilized yeast cells performed significantly better than the suspended cell culture system or batch culture. The effluent wine from the continuous fermentor system contained 7.1% (v/v) ethanol and 0.18% (w/v) residual sugar at 0.01 h^-1 dilution rate. The new continuous bioreactor system also gave 17–34 times higher maximum ethanol productivity compared to the conventional batch wine fermentation. At a low dilution rate, 0.01^-1, as high as 92% sugar to ethanol yield was achieved. Based on the results obtained from this study, the possibility of developing a continuous wine cooler fermentation process was demonstrated. A two-stage continuous wine fermentation system may be designed and operated. The grape juice can be fed into the first-stage that is operated at about 0.2 h^-1 dilution rate and the effluent from the first-stage is fed into the second-stage continuous fermentor operated at about 0.01 h^-1 dilution rate. By doing so, a wine cooler can be produced continuously and efficiently, by employing the newly designed tapered column type bioreactor charged with the immobilized yeast cells.