• Journal of Microbiology and Biotechnology
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• v.28 no.10
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• pp.1749-1759
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• 2018

Recombinant (rec) prion protein (PrP) is an extremely useful resource for studying protein misfolding and subsequent protein aggregation events. Here, we report mass production of high-purity rec-polypeptide encoding the C-terminal globular domain of PrP; (90-230) for human and (89-231) for murine PrP. These proteins were expressed as His-tagged fusion proteins in E. coli cultured by a high cell-density aerobic fermentation method. RecPrPs recovered from inclusion bodies were slowly refolded under reducing conditions. Purification was performed by a sequence of metal-affinity, cation-exchange, and reverse-phase chromatography. The current procedure yielded several dozens of milligrams of recPrP per liter with >95% purity. The purified recPrPs predominantly adopted an ${\alpha}$-helix-rich conformation and were functionally sufficient as substrates to measure the seeding activity of human and animal prions. Establishment of a procedure for high-level production of high-purity recPrP supports the advancement of in vitro investigations of PrP including diagnosis for prion diseases.

• Microbiology and Biotechnology Letters
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• v.32 no.2
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• pp.149-153
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• 2004

L-Threonine fermentation process was constructed on batch and fed-batch culture by using Escherichia coli MT201. The production type of L-threonine was observed as growth-associated production in batch culture. In fed-batch culture studying optimal concentration of yeast extract in feeding media, when 600 g/l of glucose and 60 g/l of yeast extract were added in feeding media, 87 g/$\ell$ of L-threonine was produced. To improve cell growth and L-threonine production, the culture of high cell density was performed in fed-batch culture with oxygen enriched air and feeding media containing L-methionine and phosphate. Under the conditions, we could achieve the highest L-threonine production of98 g/$\ell$ at 60 h. The highest productivity of L-threonine was about 3.85 g/$\ell$/h.

• Microbiology and Biotechnology Letters
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• v.21 no.4
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• pp.354-365
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• 1993

A novel control method with automatic tuning of PID controller parameters has been developed for efficient regulation of dissolved oxygen concentration in fed-batch fermentations of Escherichia coli. Agitation speed and oxygen partial pressure in the inlet gas stream were chosen to be the manipulated variables. A heuristic reasoning allowed improved tuning decisions from the supervision of control performance indices and it coule obviate the needs for process assumptions or disturbance patterns. The control input consisted of feedback and feedforword parts. The feedback part was determined by PID control and the feedforward part is determined from the feed rate. The proportional gain was updated on-line by a set of heuristics rules based on the supervision of three performance indices. These indices were output error covariance, the average value of output error, and input covariance, which were calculated on-line using a moving window. The integral and derivative time constants were determined from the period of output response. The specific growth rate was maintained at a low level to avoid acetic acid accumulation and thus to achieve a high cell density. The specific growthe rate was estimated from the carbon dioxide evolution rate. In fed-batch fermentation, the simutaneous control of dissolved oxygen concentration (at 0.2; fraction of saturated value) and specific growth rate (at 0.25$hr^{-1}$) was satisfactory for the entire culture period in spite of the changes in the feed rate and the switching of control input.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.268-272
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• 2003

Experiments were carried out to investigate the selection of industrial medium and optimization of fermentation process for the production of erythritol by Candida magnoliae SR101. In the batch fermentation, light steep water(LSW) was the best nitrogen source for the industrial production of erythritol. For the optimization of culture condition, the batch culture was performed. When the concentration of LSW was 65 mL/L in the defined medium containing 250 g/L of glucose, 44% of erythritol yield with 110 g/L of erythritol concentration and 0.66 g/L-hr of productivity, respectively were obtained. Two-stage fed-batch culture was performed to improve the volumetric productivity of erythritol. High density cell culture in the growth stage was performed by batch type with 100 g/L glucose and 500 mL/L LSW concentration, respectively. The cell yield was 0.72 g-cell/g-glucose. Productivity of erythritol was increased and concentration of organic acids such as gluconic acid and acetic acid were decreased when initial pH of 6.5 controlled by 28% ammonia water For increasing yield of erythritol, glucose concentration in the production stage was tested. 37% of total erythritol yield with 186 g/L of erythritol concentration and 1.66 g/L-hr of erythritol productivity were obtained when 820 g of glucose powder was directly added for making up 450 g/L of glucose at production stage.

• Korean Journal of Microbiology
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• v.37 no.1
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• pp.92-99
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• 2001

High-cell-density cultivation of Ralstonia eutopha KHB 8862 by fed-batch fermentation in a 200 l pilot plant was carried out for the mass production of poly(3-hydroxybutyrate) (PHB). After 80 h of cultivation, the dry cell weight (DCW), PHB concentration, and PHB yield from fructose syrup reached 168 g/l, 74%DCW, and 0.27 (w/w), respectively, resulting in a productivity of 1.6 g of PHB/L/h. Based on these results, the PHB production cost from bacterial fermentation was analyzed and economic evaluation was performed. In the case of new investment being implemented or not, the production cost of PHB was US$ 3.15/kg and US$ 2.41/kg, respectively. PHB productivity and PHB yield on a carbon substrate were both important factors to be optimized. The increase of PHB yield on a carbon sources significantly decreased the PHB production cost but the increase in productivity had a relatively slight effect on the decrease in PHB production cost because the cost of carbon sources (37%) for PHB was larger in proportion to total cost than the depreciation cost (17%). These results suggest that the increased PHB yield from carbon sources and the development of new cheaper substrates would be more effective in decreasing PHB production cost than the increase in productivity. It was demonstrated that PHB is not in competition with consumable plastics such as PET in present market. Therefore, it is essential to lower production cost to be used as a bulk product and desirable to develop new application fields for PHB such as biomedical and cosmeceuticals.

• Microbiology and Biotechnology Letters
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• v.44 no.3
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• pp.293-302
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• 2016

To determine the dominant bacterial species during the Saeu-jeotgal ripening process, the cultivable bacterial population was examined over a 135-day period using six different growth media. The greatest numbers of bacteria were identified when marine agar was used for culture, with maximum cell density identified at day 65 (2.51 × 10⁷ colony forming units/g). Over the course of 135 days, the bacterial diversity was analyzed eight times. A total of 467 isolates, comprising 87 species from 42 genera, as well as 16 isolates belonging to previously unknown species, were identified. The number of species detected decreased from 39 at day 1 to 13 at day 135. The order of dominance at the genus level was as follows: Staphylococcus, Salimicrobium, Kocuria, and Psychrobacter. Staphylococcus and Salimicrobium accounted for 2% of the diversity at day 1, and then increased to 39% and 36%, respectively, at day 135. The dominant species Staphylococcus equorum, Salimicrobium salexigens, and Kocuria palustris accounted for 23.6%, 16.1%, and 10.9% of all isolates, respectively. Importantly, both St. equorum and Sm. salexigens remained viable at a NaCl concentration of 21% (w/v), which indicates their strong involvement in the ripening of Saeu-jeotgal.

• KSBB Journal
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• v.11 no.6
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• pp.696-703
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• 1996

The bacterium Serratia marcescens QM Bl466 produces selectively large amount of chitinolytic enzymes(about 1mg/L medium). Enzymatic hydrolysis of chitin to N-acelyl-${\beta}$-D-glucosamine(NAG) is performed by a system consisting of two hydrolases : chitinase and chilobiase. Objectives of this study included optimization of a microbial host by using chitin particles for chitinase/chitobiase production and secretion and also development of batch fermentation system for high cell density cultivalion of S. marcescens QM B1466. Also, the influence of chitin source and carboxymethyl(CM) chitin on chitinase/chitobiase production and NAG production was investigated. When carboxymethyl chitin was substituted for colloidal and practical grade chitin, the chitinase activity was increased about 7∼10U/mL. In this case, the ratio of chitinase/chitobiase was 30.03U/3.44U(9:1). The highest amounts of NAG(3.0g/L) was obtained.

• KSBB Journal
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• v.9 no.2
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• pp.211-223
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• 1994

In fermentations with a 4-liter stirred tank bioreactor, a better than two-fold enhancement of the gas-liquid mass transfer coefficient$(k_La)$ in the celite-immobilized fungal cultures of Tolypocladium in flatum over the parallel conventional free-cell was observed at identical biomass concentrations, despite the higher specific oxygen uptake rate of the immobilized fungi during exponential growth. As a result oxygen sufficient conditions, i. e., dissolve oxygen(D.O.) concentrations exceeding 75% air saturation, could be maintained throughout exponential growth period of the immobilized culture, in contrast to the suspended fungal culture, whose D.O. levels fell below 50% air saturation. A linear monotonic dependence of $k_La$ upon impeller agitaion rate was found for both immobilized and conventional cultivation modes over a range of 250 to 550rpm, the slope being a function of biomass concentration for the free but not for the immobilized cell system In contrasts oxygen transfer rate was a much weaker function of aeration rate up to about 2.5 vvm for both culture configurations. Above this level, aeration rate had no further effect on the mass transfer. In addition, the immobilized cultures sustained good morphological and physiological states, leading to almost two times higher cyclosporln A (CyA) productivity overt the parallel free cell system. These experiments suggest that the celite-immobilized fungal system in a stirred tank reactor has considerable promise for scaling up cyclosporin A production in terms of high-density cultivation.

• Microbiology and Biotechnology Letters
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• v.49 no.4
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• pp.559-565
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• 2021

This study aimed to optimize gamma-aminobutyric acid (GABA) production by employing five strains of lactic acid bacteria (LAB) that were capable of high cell growth and GABA production using a modified synthetic medium. GABA production in the strains was qualitatively confirmed via detection of colored spots using thin layer chromatography. Lactobacillus plantarum SGL058 and Lactococcus lactis SGL027 were selected as the suitable strains for GABA production. The conditions of the carbon and nitrogen sources were determined as 5 g/l glucose (L. plantarum SGL058), 5 g/l lactose (L. lactis SGL027), 10 g/l yeast extract (L. plantarum SGL058), and 20 g/l yeast extract (L. lactis SGL027) for GABA production. The cell growth, monitored by optical density at 600 nm, was 5.93 for L. plantarum SGL058. This value was higher than the 3.04 produced by L. lactis SGL027 at 36 h using a 5-L fermenter. The highest concentration of GABA produced was 546.7 ㎍/ml by L. plantarum SGL058 and 404.6 ㎍/ml by L. lactis SGL027, representing a GABA conversion efficiency of (%, w/w) of 4.0% and 3.4%, respectively. The fermentation profiles of L. plantarum SGL058 and L. lactis SGL027 provide a basis for the utilization of LAB in GABA production using a basal synthetic medium.