• Title/Summary/Keyword: Mass culture conditions

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Optimal Culture Conditions for MK1 Strain Isolated from Soft-Rotten Tissue of Neungee Mushroom (Sarcodon aspratus) and the Physico-Chemical Properties of the Purified Exopolysaccharide of MK1 (능이버섯(Sarcodon aspratus) 무름병소에서 분리한 MK1 균주의 최적 성장조건과 정제된 균체외다당류의 특성)

  • Ryu, Jeong-Eun;Lee, Young-Nam
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.324-331
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    • 2009
  • MK1 strain, an obligate aerobic heterotrophic bacterium isolated from the rotten tissue of Neungee mushroom (Sarcodon aspratus), produces a copious amount of exopolysaccharide (EPS), which could evoke macrophage activation. Investigations on optimal culture conditions of MK1 and physical properties of MK1 EPS were made. Glucose, galactose, fructose, and sucrose supported well growth of MK1, but potato starch and dextrin did not. However, lactose seemed to be a less favorable carbon source. Optimal growth of MK1 was obtained at pH 7.0, $30^{\circ}C$, and 200 rpm with 2% glucose, and 0.2~0.05% $(NH_4)_2SO_4$. $EPS_{opt}$ obtained from an optimal growth condition constituted of carbon (37.1%), nitrogen (2.2%), oxygen (49.3%), and hydrogen (6.4%), but no sulfur. Paper chrogromatogram of the acid-hydrolysate of $EPS_{opt}$ suggested that MK1 EPS seemed to be hetropolysaccharide composed of a few number of monosaccharides including amino- and acidic-sugars. Its molecular mass determined by SDS-polyacrylamide gel electrophoresis varied from 14.8 to 47.9 kDa. Physical properties of $EPS_{glu}$ obtained from cell grown in glucose medium, such as relative viscosity ($_{rel}$) and crystalline morphology were rather affected by pH of the growth medium. Relative viscosity ($_{rel}$) of exopolysaccaride (0.1 g/ml) harvested from cells grown at medium pH ranging from 6.0 and 7.5 was 1.23 and 1.39, respectively. The freeze-dried exopolysaccharide obtained at low pH (6.0 and 6.5) was fine crystaloid and water-soluble, whereas those obtained at high pH (7.0 and 7.5) was rather gluey and less water-soluble.

Improvement of Peroxidase Productivity by Optimization of Medium Composition and Cell Inoculum Size in Suspension Cultures of Sweet Potato (Ipomoea batatas) (고구마(Ipomoea batatase)현탁배양에서 배지조성 및 세포접종량의 적정화에 의한 Pemxidase생산성 향상)

  • 곽상수;김수경;정경희;유순희;박일현;유장렬
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.2
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    • pp.91-97
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    • 1994
  • To improve the productivity of peroxidase (POD) of cell line SP-47 derived from cell suspension cultures of sweet potato (Ipomoea batatas (L) Lam.cv White Star), we optimized culture conditions including the composition and concentration of plant growth regulators and carbon source, and the cell inoculum size. When one g (fr wt) of cells was inoculated into 50 mL TL medium supplemented with l mg/L 2,4-D and 30g/L sucrose in 300 mL Erlenmeyer flask at 25$^{\circ}C$ in the dark (100rpm), the POD activity per g cell dry wt was maximized to be about 6,800 units after 25 days of subculture, which was about 30 times higher than that of intact roots of horseradish plants grown in the greenhouse, but the cell growth was maximum after 15 days of subculture. The protein content per g cell dry wt maintained almost plateau and after 25 days of subculture decreased as culture Proceeded further whereas the POD specific activity (unit/mg protein) was about two times higher after subculture and continuously increased from 12 days to the end of cultures (40 days). The POD isozyme patterns showed almost the same regardless of cell growth stage, but some acidic isozymes were slightly increased after 25 days of subculture. These results indicate that POD activity in suspension cultures of sweet potato is closely associated with cell growth and stresses derived from cell culture renditions and medium depletion. Due to its high POD activity the SPL47cell line seems to be suitable for the mass production of POD.

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Optimal conditions and effects of prebiotics for growth and antimicrobial substances production of Lactobacillus brevis BK11 (Lactobacillus brevis BK11의 증식과 항균물질 생산을 위한 최적 배양조건 및 prebiotics의 영향)

  • Lim, Eun-Seo
    • Korean Journal of Microbiology
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    • v.51 no.3
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    • pp.288-299
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    • 2015
  • Lactobacillus brevis BK11 obtained from Baikkimchi was selected to study the effects of culture medium, initial pH, atmosphere composition, incubation temperature and time, and prebiotics on growth and production of antimicrobial substances. Growth and antimicrobial substances production of L. brevis BK11 were significantly higher in MRS broth than in BHI or M17 broth. The production of cell mass, lactic acid, and bacteriocin by BK11 strain was at maximum in MRS broth adjusted to pH 6.0. Aerobic and microaerobic conditions were favored cell growth and antimicrobial substances production than anaerobic condition. Biomass and lactic acid production and antimicrobial substances activity of BK 11 were significantly better at 30 and $37^{\circ}C$ than at $25^{\circ}C$. Growth of the strain BK11 entered the stationary growth stage at 24 h after inoculation, and decreased after 36 h. Antimicrobial activities of cell-free culture supernatant and bacteriocin solution were highest when cultured in MRS broth with an initial pH 6.0 for 24-30 h at $37^{\circ}C$. In addition, the highest cell number and lactic acid and bacteriocin production were recorded in the presence of 1 and 2% (w/v) fructooligosaccharide (FOS), however, inulin and raffinose did not affect biological and physicochemical characteristics and antimicrobial activities of L. brevis BK11 cultures. According to these results, production of antimicrobial substances by L. brevis KB11 was closely associated with cell density. Under optimal conditions for antimicrobial substances production, L. brevis BK11 effectively inhibited the growth of Helicobacter pylori ATCC 43504.

Optimal Salinity and Temperature Conditions for the Growth of the Ultra-small Rotifer Synchaeta kitina (초소형 Rotifer Synchaeta kitina의 성장을 위한 최적 염분 및 수온 조건)

  • Park, Jin-Chul;Park, Heum-Gi
    • Journal of Aquaculture
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    • v.21 no.2
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    • pp.70-75
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    • 2008
  • We investigated the optimum salinity and temperature conditions for mass culture of ultra-small rotifer Synchaeta kitina. In the salinity experiment of ranging within 5 and 30 psu, the population growth of S. kitina increased continuously up to 20 psu, and then decreased over 20 psu. Their maximum density showed 390.1 inds./mL at 5 psu. A pre-reproductive phase was shortened in low salinity than high salinity. Also, the maximum offsprings and maximum lifespan and lifespan of the female were 13.4 inds. and 5.9 days, respctively at 5 psu. In the temperature experiments of ranging within 16 and $32^{\circ}C$, the population growth of S. kitina increased continuously up to $24^{\circ}C$, and then decreased over $24^{\circ}C$. The highest maximum density showed 492.8 inds./mL at $16^{\circ}C$. Their offsprings increased significantly with temperature decrease, and the maximum number of offsprings per female was 9.2 females. at $16^{\circ}C$. Their lifespan increased with temperatures decrease and the longest lifespan was to 5.5 days at $16^{\circ}C$. From these results, we conclude that the optimum culture conditions of S. kitina is 5 psu and $16^{\circ}C$.

Antifungal Activities of Pseudomonas spp. Strains Against Plant Pathogens and Optimization of Culture Conditions (식물병원성 진균에 항균 효과를 지닌 슈도모나스 균주의 항진균 활성 증진을 위한 배양조건의 최적화)

  • Chang, Seog-Won;Choi, Byung-Jin;Hong, Jeum-Kyu;Rho, Yong-Taek
    • Korean Journal of Microbiology
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    • v.46 no.3
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    • pp.248-254
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    • 2010
  • To define the optimum conditions for the mass production of four antifungal Pseudomonas spp. isolated from soil, we have investigated culture conditions and effects of various nutrient sources on the bacterial growth and evaluated antagonistic activity against Rhizoctonia solani and Sclerotinia homoeocarpa, plant pathogens. The optimum temperature and pH for the growth of these isolates were determined as pH 7.0 and $20^{\circ}$ or $25^{\circ}C$, respectively. Sucrose, tryptone, and $K_2HPO_4$ generally were more adequate for better growth as carbon, nitrogen and mineral source, respectively. The nutrient sources were also found to be very effective for high antifungal activities against R. solani and S. homoeocarpa. It was elucidated that YUD-F group (P. mandelii and P. fluorescens), which inhabit regions at relatively low temperature, had more broad spectrum and higher antifungal activity than YUD-O group (P. trivialis and P. jessenii) generally against R. solani and S. homoeocarpa. It is thought that the differences of the average temperature in the various habitats of Pseudomonas spp. influence the optimal growth temperature and antifungal activity. Especially, Pseudomonas spp. of YUD-O group showed the better antifungal activity against dollar spot caused by S. homoeocarpa, but showed relatively weaker antifungal activity against brown patch caused by R. solani.

Optimum Culture Density for the Intensive Mass Production in Cyclopoid Copepod, Paracyclopina nana (Cyclopoid 요각류, Paracyclopina nana의 대량배양을 위한 최적밀도)

  • Lee Kyun-Woo;Park Heum-Gi
    • Journal of Aquaculture
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    • v.18 no.1
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    • pp.19-25
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    • 2005
  • This study was performed to investigate the optimum density for the intensive mass production of cyclopoid copepod, Paracyclopina nana in terms of nauplii and adults production. Effect of three development stages on the fecundity of adult female for nauplii production, survival rate of P. nana nauplii with different initial nauplii culture densities for adults production and cannibalistic feeding behavior of P. nana was examined, respectively. The fecundity of adult female by different female, copepodite ana nauplii density in 2 ml water volume decreased with the density of adult female, but was not affected by the density of either copepodite or nauplii. The average daily nauplii production for a adult female in 8 L water volume was $2.3{\times}10^5$ individuals with the incubation density of 7 adult females/ml, and this average value was significantly higher than those values of 0.6 to $1.7\times10^5$ individuals with the incubation density of 1,3,5, 10 adult females/ml (P<0.05). Survival rate of P. nana nauplil with different initial nauplii culture densities in 5 L vessels for 15 days were 32.7, 30.7, 28.9 and $23.0\%$ with the culture density of 50, 100, 150 and 200 inds./ml, respectively, but these were significantly higher than those of values 19.7 and $18.4\%$ with the culture density of 250 and 300 inds./ml (P<0.05). Cannibalistic behavior of P. nana adults on their offspring was observed, but the behavior decreased when phytoplankton was supplemented though there was no statistical difference (P>0.05). These results may indicate that P. nana is adaptable to the hatchery conditions and this species is cultured at the high densities. Optimum culture density for nauplii and adults production of P. nana were 7 adult females/ml and 200 nauplii/ml, respectively.

Quantitative Proteomics Towards Understanding Life and Environment

  • Choi, Jong-Soon;Chung, Keun-Yook;Woo, Sun-Hee
    • Korean Journal of Environmental Agriculture
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    • v.25 no.4
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    • pp.371-381
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    • 2006
  • New proteomic techniques have been pioneered extensively in recent years, enabling the high-throughput and systematic analyses of cellular proteins in combination with bioinformatic tools. Furthermore, the development of such novel proteomic techniques facilitates the elucidation of the functions of proteins under stress or disease conditions, resulting in the discovery of biomarkers for responses to environmental stimuli. The ultimate objective of proteomics is targeted toward the entire proteome of life, subcellular localization biochemical activities, and the regulation thereof. Comprehensive analysis strategies of proteomics can be classified into three categories: (i) protein separation via 2-dimensional gel electrophoresis (2-DE) or liquid chromatography (LC), (ii) protein identification via either Edman sequencing or mass spectrometry (MS), and (iii) proteome quantitation. Currently, MS-based proteomics techniques have shifted from qualitative proteome analysis via 2-DE or 2D-LC coupled with off-line matrix assisted laser desorption ionization (MALDI) and on-line electrospray ionization (ESI) MS, respectively, toward quantitative proteome analysis. In vitro quantitative proteomic techniques include differential gel electrophoresis with fluorescence dyes. protein-labeling tagging with isotope-coded affinity tags, and peptide-labeling tagging with isobaric tags for relative and absolute quantitation. In addition, stable isotope-labeled amino acids can be in vivo labeled into live culture cells via metabolic incorporation. MS-based proteomics techniques extend to the detection of the phosphopeptide mapping of biologically crucial proteins, which ale associated with post-translational modification. These complementary proteomic techniques contribute to our current understanding of the manner in which life responds to differing environment.

Mass Cultivation and Secondary Metabolite Analysis of Rhodobacter capsulatus PS-2 (광합성세균 Rhodobater capsulatus PS-2의 대량배양 최적화 및 대사산물 분석)

  • Bong, Ki Moon;Kim, Jong Min;Yoo, Jae-Hong;Park, In Chul;Lee, Chul Won;Kim, Pyoung Il
    • KSBB Journal
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    • v.31 no.3
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    • pp.158-164
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    • 2016
  • Plant growth promoting (PGP) hormones, which are produced in a small quantity by bacteria, affect in plant growth and development. PGPs play an important role on the crop productivity in agricultural field. In this study, a photosynthetic bacterial strain producing the PGP was isolated from paddy soil. Bacterial isolate was gram negative, rod-shaped and motility positive. From the 16s rRNA gene sequence analysis, the isolate was identified as Rhodobacter capsulatus PS-2. The mass cultivation of R. capsulatus PS-2 was optimized by considering of the carbon, nitrogen and inorganic salt sources. Optimal medium composition was determined as Na-succinate 4.5 g, yeast extract 5 g, $K_2HPO_4$ 1 g, $MgSO_4$ 5 g, per liter. From the result of 500 L fermentation for 2 days using the optimal medium, the viable cells were $8.7{\times}10^9cfu/mL$. R. capsulatus PS-2 strain produced the carotenoid and indole-3-acetic acid (IAA). The carotenoid extraction and quantitative analysis were performed by HCl-assisting method. Total carotenoid contents from R. capsulatus PS-2 culture broth were measured as $7.02{\pm}0.04$ and $6.93{\pm}0.05mg/L$ under photoheterotrophic and chemoheterotrophic conditions, respectively. To measure the productivity of IAA, colorimetric method was employed using Salkowski reagent at optical density 535 nm. The results showed that the highest content of IAA was $197.44{\pm}5.92mg/L$ in the optimal medium supplemented with 0.3% tryptophan.

Isolation and Mycelial Cultivation Submerged of Phellinus sp. (Phellinus sp.의 분리 및 균사체의 액체배양)

  • Kang, Tae-Su;Lee, Dong-Gi;Lee, Shin-Young
    • The Korean Journal of Mycology
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    • v.25 no.4 s.83
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    • pp.257-267
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    • 1997
  • Fruit bodies similar to the Phellinus sp. residing on the mulberry were collected at Yang-yang in Kang-won-do province and one strain of Phellinus sp. was isolated from the fruit bodies. For mass production of the isolated mycelia in a submerged culture, the culture conditions, medium composition, and the effect of various culture systems on the mycelial growth, were investigated. The morphological characteristics of the fruit body were as follows: covered with blackish to black and rough, lower surface with yellowish-brown to dull-brown and smooth, 5-7 cm thick and hard woody. Also, the pure cultured mycelia showed yellowish-brown color, capability of purplish-brown pigment production on the PDA plate media, no-formation of clamp-connection, much binding branch, and enzyme activities such as laccase, tyrosinase and peroxidase. Therefore, pure cultured strain was identified to be Phellinus sp. In the flask culture, the optimum culture conditions for the mycelial production were obtained after cultivation of 8 days at inoculum level of 5%(v/v), media volume of 70 mL, 150 rpm, initial pH 6, and temperature of $30^{\circ}C$. Optimum medium composition from the response surface analysis were determined to be glucose 12.12 g/L, sucrose 12.12 g/L, yeast extract 11.15 g/L, malt extract 11.15 g/L, $KH_2PO_4$ 0.855 g/L and $CaCl_2$ 0.855 g/L. The production of the mycelia after 4 and 8 days of cultivation was 1.95 and 9.89 g/L, respectively. The maximum specific growth rate and productivity were $0.020\;hr^{-1}$ and 1.25 g/L/day, respectively. Among the three different culture systems for the growth of mycelia, the maximum mycelial dry weight of 7.5 g/L was obtained after cultivation of 4 days in the air-lift fermentor under aeration rate of 2.5 vvm. The maximum specific growth rate and productivity were $0.033\;hr^{-1}$ and 1.9 g/L/day, respectively, which were about 1.7 and 4.2 times higher than those of flask culture.

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Development of standard shoe-lasts that reflect BMI and foot characteristics of senior generation women (여성 시니어 세대 BMI와 발 특성에 따른 구두용 표준 라스트 개발)

  • Lee, Woong-Sub;Kim, Sun-Ah
    • The Research Journal of the Costume Culture
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    • v.25 no.2
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    • pp.175-191
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    • 2017
  • The purpose of this study is to analyze the body mass index(BMI) and foot characteristics of senior-generation women and to use those values to develop a standard shoe last for producing shoes that best fit the foot of senior-generation women. In order to develop a standard shoe last that reflects the BMI and foot characteristics of senior-generation women, I analyzed the morphometric characteristics of feet by conducting descriptive statistics for each item. The foot characteristics based on BMI were analyzed by conducting ANOVA and Duncan's tests via the foot's measured values. Subsequently, I developed a standard shoe last reflecting the BMI and the foot characteristics of senior-generation women by applying all those values. Senior-generation women showed significant differences among three groups based on BMI-normal weight, overweight, and obese-in all the items except for medial ball width and toe 5 angle. Through these values, I realized that as the value of the BMI becomes higher, the size of the foot becomes larger. This suggests that as a woman's weight increases or decreases, her shoes need to be replaced as well as her clothes. Deviations in the toe-side parts of the shoe could cause deformation of the foot. If someone keeps wearing shoes produced using a shoe last with a large deviation, she risks developing foot conditions like hallux valgus. This situation points to the need for shoe manufacture that utilizes a standard shoe last that reflects the BMI and foot characteristics of the senior generation.