• Title/Summary/Keyword: Optimization of cultivation

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Development of a Novel Medium with Chinese Cabbage Extract and Optimized Fermentation Conditions for the Cultivation of Leuconostoc citreum GR1 (폐배추 추출물을 이용한 Leuconostoc citreum GR1 종균 배양용 최적 배지 및 배양 조건 개발)

  • Moon, Shin-Hye;Chang, Hae-Choon;Kim, In-Cheol
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.7
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    • pp.1125-1132
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    • 2013
  • In the kimchi manufacturing process, the starter is cultured on a large-scale and needs to be supplied at a low price to kimchi factories. However, current high costs associated with the culture of lactic acid bacteria for the starter, have led to rising kimchi prices. To solve this problem, the development of a new medium for culturing lactic acid bacteria was studied. The base materials of a this novel medium consisted of Chinese cabbage extract, a carbon source, a nitrogen source, and inorganic salts. The optimal composition of this medium was determined to be 30% Chinese cabbage extract, 2% maltose, 0.25% yeast extract, and $2{\times}$ salt stock (2% sodium acetate trihydrate, 0.8% disodium hydrogen phosphate, 0.8% sodium citrate, 0.8% ammonium sulfate, 0.04% magnesium sulfate, 0.02% manganese sulfate). The newly developed medium was named MFL (medium for lactic acid bacteria). After culture for 24 hr at $30^{\circ}C$, the CFU/mL of Leuconostoc (Leuc.) citreum GR1 in MRS and MFL was $3.41{\times}10^9$ and $7.49{\times}10^9$, respectively. The number of cells in the MFL medium was 2.2 times higher than their number in the MRS media. In a scale-up process using this optimized medium, the fermentation conditions for Leuc. citreum GR1 were tested in a 2 L working volume using a 5 L jar fermentor at $30^{\circ}C$. At an impeller speed of 50 rpm (without pH control), the viable cell count was $8.60{\times}10^9$ CFU/mL. From studies on pH-stat control fermentation, the optimal pH and regulating agent was determined to be 6.8 and NaOH, respectively. At an impeller speed of 50 rpm with pH control, the viable cell count was $11.42{\times}10^9(1.14{\times}10^{10})$ CFU/mL after cultivation for 20 hr - a value was 3.34 times higher than that obtained using the MRS media in biomass production. This MFL media is expected to have economic advantages for the cultivation of Leuc. citreum GR1 as a starter for kimchi production.

Characteristic study and optimization of culture conditions for Bacillus amyloliquefaciens SRCM 100731 as probiotic resource for companion animal (Bacillus amyloliquefaciens SRCM 100731의 반려 동물용 프로바이오틱스 소재로서의 특성 규명 및 배양 조건 최적화)

  • Ryu, Myeong Seon;Yang, Hee-Jong;Jeong, Su-Ji;Seo, Ji Won;Ha, Gwangsu;Jeong, Seong-Yeop;Jeong, Do-Youn
    • Korean Journal of Microbiology
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    • v.54 no.4
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    • pp.384-397
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    • 2018
  • The aim of this study is to screen the strains of Bacillus spp. possessing safety, probiotic activity, and so on, which can be utilized as probiotic resource for using the feed and supplement food of companion animal. About 300 isolates were isolated from traditional Korean sauces, four isolates that did not have or produce the six kinds of B. cereus type vomiting and diarrhea toxin genes, ${\beta}$-hemolytic, and three kinds of carcinogenic enzymes were selected. Antibiotic gene retention, cell surface hydrophobicity, antibiotic sensitivity, and glucose utilization were analyzed for four isolates, and finally SRCM 100731 was selected. SRCM 100731 was named as Bacillus amyloliquefaciens SRCM 100731 16S rRNA sequencing analysis, and carried out optimization of cell growth for industrial applications such as pet food and feed. The effects of 14 different components on cell growth were investigated and three significant positive factors, molasses, sodium chloride, and potassium chloride were selected as the main factors based on a Plackett-Burman design. In order to find out optimal concentration on each constituent, we carried out central composite design. The predicted optimized concentrations were 7% molasses, 1.1% sodium chloride, 0.5% potassium chloride. Finally, an overall about 7-fold increase in dry cell weight yield ($12.6625{\pm}0.0658g/L$) was achieved using the optimized medium compared with the non-optimized medium ($1.8273{\pm}0.0214g/L$). This research is expected to be highly utilized in the growing pet industry by establishing optimal cultivation conditions for industrial application as well as screening Bacillus amyloliquefaciens SRCM 100731 as probiotic resource for companion animal.

Mass Screening of Lovastatin High-yielding Mutants through Statistical Optimization of Sporulation Medium and Application of Miniaturized Fungal Cell Cultures (Lovastatin 고생산성 변이주의 신속 선별을 위해 통계적 방법을 적용한 Sporulation 배지 개발 및 Miniature 배양 방법 개발)

  • Ahn, Hyun-Jung;Jeong, Yong-Seob;Kim, Pyeung-Hyeun;Chun, Gie-Taek
    • KSBB Journal
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    • v.22 no.5
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    • pp.297-304
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    • 2007
  • For large and rapid screening of high-yielding mutants of lovastatin produced by filamentous fungal cells of Aspergillus terreus, one of the most important stage is to test as large amounts of mutated strains as possible. For this purpose, we intended to develop a miniaturized cultivation method using $7m{\ell}$ culture tube instead of traditional $250m{\ell}$ flask (working volume $50m{\ell}$). For obtaining large amounts of conidiospores to be used as inoculums for miniaturized cultures, 4 components i.e., glucose, sucrose, yeast extract and $KH_2PO_4$ were intensively investigated, which had been observed to show positive effect on enhancement of spore production through Plackett-Burman design experimet. When optimum concentrations of these components that were determined through application of response surface method (RSM) based on central composite design (CCD) were used, maximum spore numbers amounting to $1.9\times10^{10}$ spores/plate were obtained, resulting in approximately 190 fold increase as compared to the commonly used PDA sporulation medium. Using the miniaturized cultures, intensive strain development programs were carried out for screening of lovastatin high-yielding as well as highly reproducible mutants. It was observed that, for maximum production of lovastatin, the producers should be activated through 'PaB' adaptation process during the early solid culture stage. In addition, they should be proliferated in condensed filamentous forms in miniaturized growth cultures, so that optimum amounts of highly active cells could be transferred to the production culture-tube as reproducible inoculums. Under these highly controlled fermentation conditions, compact-pelleted morphology of optimum size (less than 1 mm in diameter) was successfully induced in the miniaturized production cultures, which proved essential for maximal utilization of the producers' physiology leading to significantly enhanced production of lovastatin. As a result of continuous screening in the miniaturized cultures, lovastatin production levels of the 81% of the daughter cells derived from the high-yielding producers turned out to be in the range of 80%$\sim$120% of the lovastatin production level of the parallel flask cultures. These results demonstrate that the miniaturized cultivation method developed in this study is efficient high throughput system for large and rapid screening of highly stable and productive strains.

Selection of Filamentous Cyanobacteria and Optimization of Culture Condition for Recycling Waste Nutrient Solution (폐양액 활용을 위한 Filamentous Cyanobacteria의 선발 및 최적배양)

  • Yang, Jin-Chul;Chung, Hee-Kyung;Lee, Hyoung-Seok;Choi, Seung-Ju;Yun, Sang-Soon;Ahn, Ki-Sup;Sa, Tong-Min
    • Korean Journal of Soil Science and Fertilizer
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    • v.37 no.3
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    • pp.177-183
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    • 2004
  • The discharge of waste nutrient solution from greenhouse to natural ecosystem leads to the accumulation of excess nutrients that results in contamination or eutrophication. There is a need to recycle the waste nutrient solution in order to prevent the environmental hazards. The amount and kind of nutrients in waste nutrient solution might be enough to grow photosynthetic microorganisms. Hence in the present study, we examined the growth and mass cultivation of cyanobacteria in the waste nutrient solution with an objective of removing N and P and concomitantly, its mass cultivation. Four photosynthetic filamentous cyanobacteria (Anabaena HA101, HA701 and Nostoc HN601, HN701) isolated from composts and soils of the Chungnam province were used as culture strains. Among the isolates, Nostoc HN601 performed faster growth rate and higher N and P uptake in the BG-II ($NO_3{^-}$) medium when compared to those of other cyanobacterial strains. Finally, the selected isolate was tested under optimum conditions (airflow at the rate of $1L\;min^{-1}$. in 15 L reactor, initial pH 8) in waste nutrient solution from tomato hydroponic in green house condition. Results showed to remove 100% phosphate from the waste nutrient solution in the tomato hydroponics recorded over a period of 7 days. The growth rate of Nostoc HN601 was $16mg\;Chl-a\;L^{-1}$ in the waste nutrient solution from tomato hydroponics with optimum condition, whereas growth rate of Nostoc HN601 was only $9.8mg\;Chl-a\;L^{-1}$ in BG-11 media. Nitrogen fixing capacity of Nostoc HN601 was $20.9nmol\;C_2H_4\;mg^{-1}\;Chl-a\;h^{-1}$ in N-free BG-11. The total nitrogen and total phosphate concentration of Nostoc HN601 were 63.3 mg N gram dry weight $(GDW)^{-1}$ and $19.1mg\;P\;GDW^{-1}$ respectively. Collectively, cyanobacterial mass production using waste nutrient solution under green house condition might be suitable for recycling and cleaning of waste nutrient solution from hydroponic culture system. Biomass of cyanobacteria, cultivated in waste nutrient solution, could be used as biofertilizer.

Optimization of Betacyanin Production by Red Beet (Beta vulgaris L.) Hairy Root Cultures. (Red Beet의 모상근 배양을 이용한 천연색소인 Betacyanin 생산의 최적화)

  • Kim, Sun-Hee;Kim, Sung-Hoon;Lee, Jo-No;An, Sang-Wook;Kim, Kwang-Soo;Hwnag, Baik;Lee, Hyeong-Yong
    • Microbiology and Biotechnology Letters
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    • v.26 no.5
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    • pp.435-441
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    • 1998
  • Optimal conditions for the production of natural color, betacyanin were investigated by varying light intensity, C/N ratio, concentrations of phosphate and kinds of elicitors. Batch cultivation was employed to characterize cell growth and betacyanin production of 32 days. The maximum specific growth rate, ${\mu}$$\sub$max/, was 0.3 (1/day) for batch cultivation. The maximum specific production rate, q$\^$max/$\sub$p/, was enhanced 0.11 (mg/g-cell/day) at 3 klux. A light intensity of 3 klux was shown to the best for both cell growth and betacyanin production. The maximum specific production rate was 0.125 (mg/g-cell/day) at 0.242 (1/day), the maximum specific growth rate. The dependence of specific growth rate on the light lintensity is fit to the photoinhibition model. The correlation between ${\mu}$ and q$\sub$p/ showed that the product formation parameters, ${\alpha}$ and ${\beta}$$\sub$p/ were 0.3756 (mg/cell) and 0.001 (mg/g-cell/day), respectively. The betacyanin production was partially cell growth related process, which is different from the production of a typical product in plant cell cultures. In C/N ratio experiment, high carbon concentration, 42.1 (w/w) improved cell growth rate while lower concentration, 31.6 (w/w) increased the betacyanin production rate. The ${\mu}$$\sub$max/ and q$\^$max/$\sub$p/ were 0.26 (1/day) and 0.075 (mg/g-cell/day), respectively. Beta vulgaris L. cells under 1.25 mM phosphate concentration produced 10.15 mg/L betacyanin with 13.46 (g-dry wt./L) of maximum cell density. The production of betacyanin was elongated by adding 0.1 ${\mu}$M of kinetin. This also increased the cell growth. Optimum culture conditions of light intensity, C/N, phosphate concentration were obtained as 5.5 klux, 27 (w/w), 1.25 mM, respectively by the response surface methodology. The maximum cell density, X$\sub$max/, and maximum production, P$\sub$max/, in optimized conditions were 16 (g-dry wt./L), 12.5 (mg/L) which were higher than 8 (g-dry wt./L), 4.48 (mg/L) in normal conditions. The ${\mu}$$\sub$max/ and q$\^$max/$\sub$p/ were 0.376 (1/day) and 0.134 (mg/g-cell/day) at the optimal condition. The overall results may be useful in scaling up hairy root cell culture system for commercial production of betacyanin.

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Improvement of Energy Efficiency of Plants Factory by Arranging Air Circulation Fan and Air Flow Control Based on CFD (CFD 기반의 순환 팬 배치 및 유속조절에 의한 식물공장의 에너지 효율 향상)

  • Moon, Seung-Mi;Kwon, Sook-Youn;Lim, Jae-Hyun
    • Journal of Internet Computing and Services
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    • v.16 no.1
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    • pp.57-65
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    • 2015
  • As information technology fusion is accelerated, the researches to improve the quality and productivity of crops inside a plant factory actively progress. Advanced growth environment management technology that can provide thermal environment and air flow suited to the growth of crops and considering the characteristics inside a facility is necessary to maximize productivity inside a plant factory. Currently running plant factories are designed to rely on experience or personal judgment; hence, design and operation technology specific to plant factories are not established, inherently producing problems such as uneven crop production due to the deviation of temperature and air flow and additional increases in energy consumption after prolonged cultivation. The optimization process has to be set up in advance for the arrangement of air flow devices and operation technology using computational fluid dynamics (CFD) during the design stage of a facility for plant factories to resolve the problems. In this study, the optimum arrangement and air flow of air circulation fans were investigated to save energy while minimizing temperature deviation at each point inside a plant factory using CFD. The condition for simulation was categorized into a total of 12 types according to installation location, quantity, and air flow changes in air circulation fans. Also, the variables of boundary conditions for simulation were set in the same level. The analysis results for each case showed that an average temperature of 296.33K matching with a set temperature and average air flow velocity of 0.51m/s suiting plant growth were well-maintained under Case 4 condition wherein two sets of air circulation fans were installed at the upper part of plant cultivation beds. Further, control of air circulation fan set under Case D yielded the most excellent results from Case D-3 conditions wherein air velocity at the outlet was adjusted to 2.9m/s.

IN-LINE NIR SPECTROSCOPY AS A TOOL FOR THE CONTROL OF FERMENTATION PROCESSES IN THE FERMENTED MEATS INDUSTRY

  • Tamburini, Elena;Vaccari, Giuseppe;Tosi, Simona;Trilli, Antonio
    • Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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    • 2001.06a
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    • pp.3104-3104
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    • 2001
  • The research described here was undertaken with the aim of monitoring, optimizing and ultimately controlling the production of heterofermentative microbes used as starters in the salami industry. The use of starter cultures in the fermented meats industry is a well-established technique used to shorten and standardize the ripening process, and to improve and control the organoleptic quality of the final product. Starter cultures are obtained by the submerged cultivation of suitable microorganisms in stirred, and sometimes aerated, fermenters where monitoring of key physiological parameters such as the concentration of biomass, substrates and metabolites suffers from the general lack of real-time measurement techniques applicable to aseptic processes. In this respect, the results of the present work are relevant to all submerged fermentation processes. Previous work on the application of on-line NIR spectroscopy to the lactic acid fermentation (Dosi et al. - Monreal NIR1995) had successfully used a system based on a measuring cell included in a circulation loop external to the fermenter. The fluid handling and sterility problems inherent in an external circulation system prompted us to explore the use of an in-line system where the NIR probe is immersed in the culture and is thus exposed to the hydrodynamic conditions of the stirred and aerated fluid. Aeration was expected to be a potential source of problems in view of the possible interference of air bubbles with the measurement device. The experimental set-up was based on an in-situ sterilizable NIR probe connected to the instrument by means of an optical fiber bundle. Preliminary work was carried out to identify and control potential interferences with the measurement, in particular the varying hydrodynamic conditions prevailing at the probe tip. We were successful in defining the operating conditions of the fermenter and the geometrical parameters of the probe (flow path, positioning, etc.) were the NIR readings were reliable and reproducible. The system thus defined was then used to construct and validate calibration curves for tile concentration of biomass, carbon source and major metabolites of two different microorganisms used as salami starters. Real-time measurement of such parameters coupled with the direct interfacing of the NIR instrument with the PC-based measurement and control system of the fermenter enabled the development of automated strategies for the interactive optimization of the starter production process.

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Production of gamma-Aminobutyric Acid (GABA) by Lactobacillus plantarum subsp. plantarum B-134 Isolated from Makgeolli, Traditional Korean Rice Wine (한국전통주인 막걸리로부터 분리한 Lactobacillus plantarum subsp. plantarum B-134의 gamma-aminobutyric acid (GABA)의 생산)

  • Lee, Hyun-Ju;Son, Jae-Young;Lee, Sang-Jae;Lee, Han-Seung;Lee, Bae-Jin;Choi, In-Soon;Sohn, Jae Hak
    • Journal of Life Science
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    • v.27 no.5
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    • pp.567-574
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    • 2017
  • This study is to isolate and identify ${\gamma}$-amino butyric acid (GABA) producing lactic acid bacteria (LAB) from Makgeolii, traditional Korean rice wine and then establish the optimal culture conditions for GABA production. Sixty four LAB from Makgeolli were isolated according to the characteristics of the shape and color of the colony grown on MRS agar plate. The GABA production of the isolated strain cultured in MRS broth contained 1% MSG (mono-sodium glutamate) were determined and evaluated by TLC and HPLC analysis. Strain B-134 was selected for highest GABA production. From the analysis of 16S rRNA and glutamate decarboxylase B (gadB) gene sequences, strain B-134 was tentatively identified as a Lactobacillus plantarum subsp. plantarum B-134. Effects of culture parameters, including glutamic acid level, culture temperature, NaCl level, and pH on GABA production were investigated for culture optimization. The optimum culture condition for GABA production by B-134 were culture temperature of $37^{\circ}C$, pH of 5.7, NaCl content of 0% (w/v) and MSG content of 3% (w/v), which produced 25 mM of GABA during cultivation time of 48 hr. From these results, strain B-134 is expected to be utilized as useful microorganisms for GABA-enriched health beneficial food.

Development of Life Science and Biotechnology by Marine Microorganisms (해양 미생물을 활용한 생명과학 및 생명공학 기술 개발)

  • Yongjoon Yoon;Bohyun Yun;Sungmin Hwang;Ki Hwan Moon
    • Journal of Life Science
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    • v.33 no.7
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    • pp.593-604
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    • 2023
  • The ocean accounts for over 70% of the Earth's surface and is a space of largely unexplored unknowns and opportunities. Korea is a peninsula surrounded by the sea on three sides, emphasizing the importance of marine research. The ocean has an extremely complex environment with immense biological diversity. In terms of microbiology, the marine environment has varying factors like extreme temperature, pressure, solar radiation, salt concentration, and pH, providing ecologically unique habitats. Due to this variety, marine organisms have very different phylogenetic classifications compared with terrestrial organisms. Although various microorganisms inhabit the ocean, studies on the diversity, isolation, and cultivation of marine microorganisms and the secondary metabolites they produce are still insufficient. Research on bioactive substances from marine microorganisms, which were rarely studied until the 1990s, has accelerated in terms of natural products from marine Actinomycetes since the 2000s. Since then, industries for bioplastic and biofuel production, carbon dioxide capture, probiotics, and pharmaceutical discovery and development of antibacterial, anticancer, antioxidant, and anti-inflammatory drugs using bacteria, archaea, and algae have significantly grown. In this review, we introduce current research findings and the latest trends in life science and biotechnology using marine microorganisms. Through this article, we hope to create consumer awareness of the importance of basic and applied research in various natural product-related discovery fields other than conventional pharmaceutical drug discovery. The article aims to suggest pathways that may boost research on the optimization and application of future marine-derived materials.

Optimization of Culture Condition for Enhancing the Probiotics Functions (프로바이오틱스의 기능성 향상을 위한 배양법)

  • Chang, Bo Yoon;Han, Ji Hye;Cha, Bum-Suk;Ann, Sung-Ho;Kim, Sung Yeon
    • Journal of Food Hygiene and Safety
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    • v.30 no.3
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    • pp.295-301
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    • 2015
  • The functions of probiotics, particularly Lactic acid bacteria, have been studied in a range of human diseases, including cancer, infectious diseases, gastrointestinal disorders, and allergies. Among the many benefits associated with the consumption of probiotics, modulation of immune activity has received the most attention. This study aimed at investigating the improved immune stimulatory and stability of L. plantarum when cultivated on modified basal media supplemented with the Undaria pinnatifida co-cultured with L. plantarum. An in vitro test showed that U. pinnatifida media cultured L. plantarum is strong enough to survive in the gastric juice (gastric and bile acid). Mouse macrophage-derived cell lines RAW 264.7 was used to measured immune stimulating activity of L. plantarum. When U. pinnatifida media cultured by L. plantarum was NO and $TNF-{\alpha}$ production is significantly increased compared to basal media cultured L. plantarum. These results show that U. pinnatifida could be applied for a component for cultivation of L. plantarum. This optimized U. pinnatifida medium can be used the improving of stability and immune function on production of probiotics.