• Membrane Journal
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• v.2 no.2
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• pp.122-128
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• 1992

A study on the citric acid production was performed in various size dual hollow fiber bioreactors with immobilized Aspergillus niger (KCTC 1232). The final dry cell mass density reached 300g/l based on the space volume available for cell growth. Under air and oxygen aeration the volumethe productivity reached 0.63 and 1.02g/l.h, which cormsponded to 10 and 16 fold over those of batch fermentation, respectively. The initial pH of the medium was a critical factor and the lower value resulted in higher citric acid yield. The increase in the feeding rate of medium or the number of reactor unit resulted in the improvement of the productivity due to higher consumption rate of substrate.

• Journal of Microbiology and Biotechnology
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• v.25 no.12
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• pp.2058-2071
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• 2015

A comparative study was conducted to evaluate precision and accuracy in controlling the temperature dependence of encapsulated microbial time-temperature integrators (TTIs) developed using two different emulsification techniques. Weissela cibaria CIFP 009 cells, immobilized within 2% Na-alginate gel microbeads using homogenization (5,000, 7,000, and 10,000 rpm) and Shirasu porous glass (SPG) membrane technologies (10 μm), were applied to microbial TTIs. The prepared micobeads were characterized with respect to their size, size distribution, shape and morphology, entrapment efficiency, and bead production yield. Additionally, fermentation process parameters including growth rate were investigated. The TTI responses (changes in pH and titratable acidity (TA)) were evaluated as a function of temperature (20℃, 25℃, and 30℃). In comparison with conventional methods, SPG membrane technology was able not only to produce highly uniform, small-sized beads with the narrowest size distribution, but also the bead production yield was found to be nearly 3.0 to 4.5 times higher. However, among the TTIs produced using the homogenization technique, poor linearity (R²) in terms of TA was observed for the 5,000 and 7,000 rpm treatments. Consequently, microbeads produced by the SPG membrane and by homogenization at 10,000 rpm were selected for adjusting the temperature dependence. The E_a values of TTIs containing 0.5, 1.0, and 1.5 g microbeads, prepared by SPG membrane and conventional methods, were estimated to be 86.0, 83.5, and 76.6 kJ/mol, and 85.5, 73.5, and 62.2 kJ/mol, respectively. Therefore, microbial TTIs developed using SPG membrane technology are much more efficient in controlling temperature dependence.

• Microbiology and Biotechnology Letters
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• v.15 no.6
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• pp.430-435
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• 1987

A chitin-immobilized enzyme amyloglucosidase(AMG) and a bacterium Zymomonas mobilis were coentrapped in alginate gel beads. Ethanol production was performed in a packed bed column reactor in a simultaneous saccharification and fermentation(SSF) mode using liquefied sago starch as a substrate. It was found that this process eliminated product inhibition and reverse reaction of glucose enhancing the rate of saccharification and ethanol production. At a low dilution rate of D = 0.11 hr$^{-1}$, the steady-state ethanol concentration was 46.0g/$m\ell$ (96.8 % of theoretical yield). The maximum ethanol productivity was 17.7g/$m\ell$, h at D = 0.83 hr$^{-1}$ when the calculation was based on the total working volume. The continuous production of ethanol was maintained stably over 40 days without problems in this reactor system.

• Analytical Science and Technology
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• v.12 no.3
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• pp.218-223
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• 1999

A single use, screen-printed sensor for the measurement of liquid phase ethanol was developed and its electrochemical performance was investigated. Disposable type edthanol sensor was fabricated by serially screen printing the carbon paste, silverd pasted and insulator inlon a polyester substrate to pattern working and reference electrode sites and electrical contact. Alcohol dehydrogenase(ADH) or alcohol oxidase(AOD) together with appropriate electron transfer mediators was immobilized on the working electrode. To improve the sensitivity and reproducibility of carbon paste electrode, some pretreatment procedures were applied and their resultant electrochemical performance was examined. The disposable type electrochemical ethanol sensor developed in this study conveniently determines the ethanol in liquid samples such as blood and in fermentation process.

• Journal of Microbiology and Biotechnology
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• v.23 no.7
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• pp.942-952
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• 2013

Monoliths are functional porous materials with a three-dimensional continuous interconnected pore structure in a single piece. A monolith with uniform shape based on poly(${\gamma}$-glutamic acid) (PGA) has been prepared via a thermally induced phase separation technique using a mixture of dimethyl sulfoxide, water, and ethanol as solvent. The morphology of the obtained monolith was observed by scanning electron microscopy and the surface area of the monolith was evaluated by the Brunauer Emmett Teller method. The effects of fabrication parameters such as the concentration and molecular mass of PGA and the solvent composition have been systematically investigated. The PGA monolith was cross-linked with hexamethylene diisocyanate to produce the water-insoluble monolith. The addition of sodium chloride to the phase separation solvent affected the properties of the cross-linked monolith. The swelling ratio of the cross-linked monolith toward aqueous solutions depended on the buffer pH as well as the monolith fabrication condition. Copper(II) ion was efficiently adsorbed on the cross-linked PGA monolith, and the obtained copper-immobilized monolith showed strong antibacterial activity for Escherichia coli. By combination of the characteristic properties of PGA (e.g., high biocompatibility and biodegradability) and the unique features of monoliths (e.g., through-pore structure, large surface area, and high porosity with small pore size), the PGA monolith possesses large potentials for various industrial applications in the biomedical, environmental, analytical, and separation fields.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1979.04a
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• pp.113.2-114
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• 1979

$\beta-Galactosidase$ is an enzyme which catalizes hydrolysis of lactose, a natural substrate, to glucose and galctose and transferring some monosac-charide units to active acceptors as sugar or alcohol. The occurence of $\beta-Galactosidase$ is known in various microorganisms, animals and higher plants and has been studied by many investigatigators. Especially, a great deal of articles for the enzyme of E. coli have been presented in genetic control mechanism and induction-repression effects of proteins, On the other hand, in the dairly products industry, it is important to hydrolyes lactosd which is the principal sugar of milk and milk products. During the last few years, the interest in enzymatic hydrolysis of milk lactose has teen increased, because of the lactose intolerence in large groups of the population. Microbial $\beta-Galactosidases$ are considered potentially most suitable for processing milk to hydrolyse lactose and, in recent years, the immobilized enzyme from yeast has been examined. Howev, most of the microbial $\beta-Gal$ actosidase are intracellular enzymes, except a few fungal $\beta-Gala-$ ctosidases, and extracellular $\beta-Galactosidase$ which may be favorable to industrial applieation is not so well investigated. On this studies, a mold producing a potent extracellular $\beta-Galactosidase$ was isolated from soil and identified as an imperfect fungus, Beauveria bassians. In this strain, both extracellular and intracellular $\beta-Galactosidases$ were produced simultaneously and a great increase of the extracellular production was acheved by improving the cultural conditions. The extracellular enzyme was purified more than 1, 000 times by procedures including Phosphocellulose and Sephadex G-200 chromatographies. Several characteristics of the enzymewas clarified with this preparation. The enzyme has a main subunit of molecular weight of 80, 000 which makes an active aggregate. And at neutral pH range, it has optimum pH for activity and stability. The Km value was determined to be 0.45$\times$10$^{-3}$ M for $o-Nitrophenyl-\beta-Galactoside.$ In any event, it is interesting to sttudy the $\beta-Galactosidase$ of B. bassiana for the mechanism of secretion and conformational structure of enzyme.

• Korean Journal of Food Science and Technology
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• v.35 no.1
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• pp.103-110
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• 2003

Stable production of fermented kanjang containing 1.8% (v/v) ethanol was obtained within four days using traditional kanjang containing 4% added glucose in packed-bed bioreactor systems filled with immobilized Zygosaccharomyces rouxii and Candida versatilis on porous alumina ceramic bead carrier at $28{\pm}0.5^{\circ}C$ and aeration rate of 0.05 vvm. Specific rates of alcohol production for Z. rouxii and C. versatilis were 0.0033 and 0.0031/day, respectively, and those of glucose consumption were both -0.0087/day in the batch type of alcoholic fermentation. In semi-continuous alcoholic fermentation at a dilution rate of 0.25/day, specific rates of alcohol production for Z. rouxii and C. versatilis were 0.0045 and 0.0029/day, and those of glucose consumption were -0.01 and -0.008/day, respectively, using identical bioreactor system. Similar specific rates of alcohol production were observed both in the batch or semi-continuous process and in the continuous one at the dilution rate of 0.25/day. Sensory characteristics of all alcoholic-fermented kanjang by Z. rouxii, C. versatilis, and a mixture of both yeasts (2:1, w/w) were shown to be significantly superior to those of home-made kanjang as revealed through organoleptic evaluation tests (p<0.05).

• Korean Journal of Food Science and Technology
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• v.30 no.2
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• pp.425-433
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• 1998

The characteristics of the reaction of calcium carbonate $(CaCO_3)$ immobilized with alginate as buffer system for the high concentration cultivation of bifidobacteria in fermenter are described by the mathematical model, and tested for the reusing possibility of the used $CaCO_3$ beads. When$CaCO_3$ beads with the various diameters were reacted in 0.1 M of the mixed organic acids (0.6 M of acetic acid and 0.4 M lactic acid) and in fermenter inoculated Bifidobacterium longum ATCC 15707, the change of bead diameters can be calculated with the amount of the decreased $CaCO_3$ from the surface of bead using the mathematical model. These values was similar to the directly measured bead diameter by a micrometer. Therefore, it was considered that the mathematical model could be used for explaining the reaction charateristics of the $CaCO_3$ bead reacted with the organic acids. When Bifidobacterium longum was incubated at $37^{\circ}C$ for 20 hours in fermenter with $CaCO_3$ beads, the buffering effect of $CaCO_3$, the reduce rate of the bead diameter, and the growth rate of Bifidobacterium longum were higher at the smaller beads than beads with the larger diameters. Also, when Bifidobacterium longum was incubated in fermenter with the mixed beads which were added new beads to the recovered beads in order to equalize with the total surface area of initial beads, the buffering effect of $CaCO_3$ bead and the growth rate of Bifidobacterium longum were very corresponded with the results of the fermentation using the only initial beads. Therfore, it is expected that the used beads can be reused by adding the initial beads.

• Korean Journal of Food Science and Technology
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• v.21 no.1
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• pp.154-163
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• 1989

This studies were designed to improve the productivity of L-lysine by protoplast fusion and immobilized system of fusants using strains of Brevibacterium flavum ATCC 21528, Brevibacterium lactofermentum ATCC 21086 and Corynebacterium glutamicum 820. Mutants were isolated with concentration method of $300{\mu}g/ml$ penicillin-G after treatment of $250{\mu}g/ml$ N-methyl-N-nitro-N-nitrosoguanidine. B. flavum $37-2(Hos^-,\;Kan^r,\;AEC^r)$, B. lactofermentum $6-2(Ile^-,\;Val^-,\;Str^r,\;AEC^r)$ and C. glutamicum 57-5$(Met^-,\;Thr^-,\;Rif^r,\;AEC^r)$ were isolated from mutants. Protoplasts were induced by being incubated with $500{\mu}g/ml$ lysozyme of lysis solution for 6 hr and the ratio of protoplast formation and regeneration were ranging from 97-99% and 33-37%, respectively. Fusion frequencies of fusants of BBFL 21, BCFG 37 and BCLG 59 were shown in the range from $1.25{\times}10^{-6}\;to\;5.83{\times}10^{-7}$ under the optimum conditions. The fusant BBFL 21 showed the highest productivity of $411.1\;ng/ml{\cdot}hr$ L-lysine in the lysine productivity broth at $30^{\circ}C$ for 72hr. In the immobilization systems, fusant BBFL 21 was employed in various polymer matrices such as sodium alginate, polyacrylamide, agar and ${\alpha}-carrageena$. The immobilization of sodium alginate showed the highest productivity of $413\;ng/ml{\cdot}hr$ L-lysine in the batch system. Continuous fermentation of immobilization system by using tube fermentor was produced the highest productivity $416.7\;ng/ml{\cdot}hr $ L-lysine under optimum condition.

• Microbiology and Biotechnology Letters
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• v.9 no.3
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• pp.159-164
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• 1981

Reinforced Calcium-alginate gel entrappment method for enzyme immobilization is described with an example of penicillin amidase from Bacillus megaterium KFCC 10029, a partially constitutive mutant of B. megaterium ATCC 14945. Penicillin amidase recovered from the fermentation broth by adsorption on celite is mixed with alginate and gelatin solution, and cast into a pellet or noodle form by coagulation in calcium salt solution followed by crosslinking with glutaraldehyde. Optimum pH and temperature of the immobilized enzyme preparation were 8.0 and 6$0^{\circ}C$, respectively. Kinetic constants such as Km value and the inhibition constant of 6-APA and phenylacetic acid were 2.6 mM, 7.4 mM and 21.2 mM, respectively. The enzyme leakage from the adsorbent during operation was successfully prevented owing to the increase of physical strength of gel coat. The half lives in a column reactor were 6 and 30 days at the respective temperature of 4$0^{\circ}C$ and 3$0^{\circ}C$, which were the 6-8 fold increased values as compared with those of without entrappment. The results highly recommended the use of reinforced Calcium-alginate gel entrappment method for the enhancement of physical strength and the operational stability of alginate gel entrapped enzyme.