• Title/Summary/Keyword: fed-batch growth

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Effect of Ca^{++} and Cu^{++} Removal from Molasses on Yest cell Growth and RNA Accumulation. (당밀로부터 Ca^{++} 및 Cu^{++} 이온 제거가 효모 생육 및 RNA축적에 미치는 영향)

  • 김재범;허선연;김중균;남희섭;남수완
    • Microbiology and Biotechnology Letters
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    • v.31 no.3
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    • pp.211-215
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    • 2003
  • When Saccharomyces cerevisiae MTY62, a high-RNA content yeast, was cultivated by fed-batch mode feeding molasses and com steep liquor, the cell density less than 45g-DCW/L and the RNA content less than 140mg/g-cell were obtained, indicating that unknown compounds inhibiting the cell growth and RNA accumulation are contained in the molasses. Therefore, in order to obtain higher levels of cell density and RNA content, $Ca^{++}$, $Cu^{++}$and $K^{+}$ ions in molasses were removed by pretreatments of molasses with various agents such as IonClear BigBead, $Na_2$$HPO_4$, $H_2$$SO_4$, citric acid, $K_2$$HPO_4$, and EDT A. Among them, IonClear BigBead, $Na_2$$HPO_4$, and EDTA gave the highest $Ca^{++}$ removal efficiency of about 60-90%. In the batch culture with pretreated molasses, the cell concentration of 18.6g-DCW/I and RNA concentration of 3127 mg/I, maximum specific growth rate of 0.459$h^{-1}$ , and specific consumption rate of reducing sugar of 1.28g-sugar/g-cell-h were obtained, which are about 10%, 17%,47%, and 36% higher levels, respectively, over the batch culture with untreated molasses.

Production of the Fungal Lipid Containing ${\gamma}-Linolenic$ Acid from Mucor sp. KCTC 8405P (Mucor sp. KCTC 8405P에 의한 ${\gamma}-Linolenic$ Acid 함유 곰팡이 유지의 생산)

  • Park, Jong-Hyun;Shin, Hyun-Kyung
    • Korean Journal of Food Science and Technology
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    • v.24 no.4
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    • pp.326-329
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    • 1992
  • Mucor sp. KCTC 8405P was cultivated in a jar fermentor for the production of fungal lipid containing ${\gamma}-linolenic$ acid with feeding the glucose solution periodically. The transition of the fungal growth into the mycelial phase from yeast-like growth was achieved by pH shift after the first two day of cultivation in the low pH medium and then lipid accumulation was accelerated until the seven day of cultivation, when the glucose in the culture broth was almost consumed. With the culture conditions applied in this experiment, biomass of 99.3 g/l by the dry cell weight and the total extractable lipid of 38.0 g containing 3.5 g/l ${\gamma}-linolenic$ acid were obtained.

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In vitro and in vivo evaluation of kenaf (Hibiscus cannabinus L.) as a roughage source for beef cattle

  • Oh, Seongjin;Mbiriri, David Tinotenda;Ryu, Chaehwa;Lee, Kangheon;Cho, Sangbuem;Choi, Nag-Jin
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.10
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    • pp.1598-1603
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    • 2018
  • Objective: The goal of this study was to evaluate kenaf as a roughage source in vitro and its effects on meat quality of Hanwoo (Korean native) cattle. Methods: Three roughage materials, rice straw silage, ryegrass silage, and kenaf silage, were tested in a batch culture and feeding trial. Rumen fermentation parameters, including gas, pH, volatile fatty acid (VFA), and ammonia were analyzed. In the feeding trial, Hanwoo steers ($373.5{\pm}5.1kg$, n = 36, 11 month of age) were divided into three feeding groups (n = 12 each). Animals were fed with each silage and concentrate until the fattening stage. Results: Crude protein, ether extract, and non-structural carbohydrates were greater in kenaf silage. Total gas production was higher in ryegrass silage, followed by kenaf silage and rice straw silage (p<0.05). Total VFA and individual VFA (acetate, propionate, and n-butyrate) were greater in kenaf silage than rice straw silage (p<0.05). In vitro dry matter digestibility showed a similar trend to that of total gas and VFA production; it was higher in ryegrass silage and lower in rice straw (p<0.05). Throughout the feeding trial, the rice straw silage group showed significantly greater average daily gain than did the others (p<0.05). The feed conversion ratio in the group fed kenaf silage was significantly greater than that of others (p<0.05). No significant differences were observed in yield or quality traits, including carcass weight, ribeye area, backfat thickness, and scores for marbling, meat color, and fat color (p>0.05). Conclusion: The results indicated that no negative effects on growth performance and carcass characteristics occurred across treatments. Therefore, kenaf could be substituted for rice straw, which is most widely used as a roughage source in Korea.

Selection of plant oil as a supplemental energy source by monitoring rumen profiles and its dietary application in Thai crossbred beef cattle

  • Matsuba, Keiji;Padlom, Apirada;Khongpradit, Anchalee;Boonsaen, Phoompong;Thirawong, Prayad;Sawanon, Suriya;Suzuki, Yutaka;Koike, Satoshi;Kobayashi, Yasuo
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.10
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    • pp.1511-1520
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    • 2019
  • Objective: The present study was conducted to select a plant oil without inhibitory effects on rumen fermentation and microbes, and to determine the optimal supplementation level of the selected oil in a series of in vitro studies for dietary application. Then, the selected oil was evaluated in a feeding study using Thai crossbred beef cattle by monitoring growth, carcass, blood and rumen characteristics. Methods: Rumen fluid was incubated with substrates containing one of three different types of plant oil (coconut oil, palm oil, and soybean oil) widely available in Thailand. The effects of each oil on rumen fermentation and microbes were monitored and the oil without a negative influence on rumen parameters was selected. Then, the dose-response of rumen parameters to various levels of the selected palm oil was monitored to determine a suitable supplementation level. Finally, an 8-month feeding experiment with the diet supplemented with palm oil was carried out using 12 Thai crossbred beef cattle to monitor growth, carcass, rumen and blood profiles. Results: Batch culture studies revealed that coconut and soybean oils inhibited the most potent rumen cellulolytic bacterium Fibrobacter succinogenes, while palm oil had no such negative effect on this and on rumen fermentation products at 5% or higher supplementation level. Cattle fed the diet supplemented with 2.5% palm oil showed improved feed conversion ratio (FCR) without any adverse effects on rumen fermentation. Palm oil-supplemented diet increased blood cholesterol levels, suggesting a higher energy status of the experimental cattle. Conclusion: Palm oil had no negative effects on rumen fermentation and microbes when supplemented at levels up to 5% in vitro. Thai crossbred cattle fed the palm oil-supplemented diet showed improved FCR without apparent changes of rumen and carcass characteristics, but with elevated blood cholesterol levels. Therefore, palm oil can be used as a beneficial energy source.

Development of an Alcoholic Drink Using Onion Extract. (양파즙을 사용한 알코올 음료의 개발)

  • Kim, Sam-Woong;Oh, Eun-Hye;Jun, Hong-Ki
    • Journal of Life Science
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    • v.18 no.7
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    • pp.980-985
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    • 2008
  • This study was carried out to develope an alcoholic drink by fermentation of onion extract using Saccharomyces cerevisiae. The optimal conditions for ethanol production were obtained by standing culture at $25^{\circ}C$ for 5 days with 5% inoculum volume. At the results by flask culture, the growth curve of used S. cerevisiae reached to the stantionary phase at 48 hr and the death phase at 90 hr, whereas ethanol production reached maximum at 114 hr. Under the above conditions, a large scale production was carried out. A standing culture in 5 l fermenter showed the similar results to its flask culture, but progressed 24 hr rapidly more than that of the flask culture. A fed-batch culture was performed by addition of the onionic medium supplemented with 10% (v/v) sucrose after 72 hr from the fermenting start. The fed-batch culture could prevent S. cerevisiae from entering into the death phase and maintain constant level of alcohol production. A continuous culture was able to carry out by adding per every 24 hr the onionic medium supplemented with 10% (v/v) sucrose after 72 hr from the fermenting start. Although S. cerevisiae used showed a little decreased growth, alcohol production maintained roughly the constant level at the maximum yield. To enhance the quality of this alcoholic drink, $2-O-{\alpha}-D-glucopyranosyl$ L-ascorbic acid (AA-2G) was supplemented into the onion extract of the substrate for fermentation. As resulted at this study, this alcoholic drink containing AA-2G should be used as a functional fermented alcohol drink strengthened with vitamin C.

Fermentation Process Development of Recombinant Hansenula polymorpha for Gamma-Linolenic Acid Production

  • Khongto, B.;Laoteng, K.;Tongta, A.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.11
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    • pp.1555-1562
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    • 2010
  • Development of the strain and the fermentation process of Hansenula polymorpha was implemented for the production of ${\gamma}$-linolenic acid ($GLA,\;C18:3{\Delta}^{6,9,12}$), an n-6 polyunsaturated fatty acid (PUFA) that has been reported to possess a number of health benefits. The mutated ${\Delta}^6$-desaturase (S213A) gene of Mucor rouxii was expressed in H. polymorpha under the control of the methanol oxidase (MOX) promoter. Without the utilization of methanol, a high-cell-density culture of the yeast recombinant carrying the ${\Delta}^6$-desaturase gene was then achieved by fed-batch fermentation under glycerol-limited conditions. As a result, high levels of the ${\Delta}^6$-desaturated products, octadecadienoic acid ($C18:2{\Delta}^{6,9}$), GLA, and stearidonic acid ($C18:4{\Delta}^{6,9,12,15}$), were accumulated under the derepression conditions. The GLA production was also optimized by adjusting the specific growth rate. The results show that the specific growth rate affected both the lipid content and the fatty acid composition of the GLA-producing recombinant. Among the various specific growth rates tested, the highest GLA concentration of 697 mg/l was obtained in the culture with a specific growth rate of 0.08 /h. Interestingly, the fatty acid profile of the yeast recombinant bearing the Mucor ${\Delta}^6$-desaturase gene was similar to that of blackcurrant oil, with both containing similar proportions of n-3 and n-6 essential fatty acids.

Identification of Pseudomonas fluorescens antagonistic to Pseudomonas tolaasii and its cultivation (버섯의 갈변병 유발세균 Pseudomonas tolaasii의 길항세균인 Pseudomonas fluorescens의 분리동정 및 배양조건)

  • 박범식;조남철전억한
    • KSBB Journal
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    • v.7 no.4
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    • pp.296-301
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    • 1992
  • A Pseudomonas fluorescens was selected from mushrooms and studied in both batch and fed-batch cultures in order to get maximal biomass concentration. P. fluorescens is an aerobic bacterium and antagonistic to Pseudomonas tolaasii which causes blotch disease on the mushroom cap. P fluarescens and P. tolaasii were identified by Gram staining, gelatin liquefaction, oxidase test, etc. and were characterized by pigment production, temperature sensitivity, salt tolerance and rapid pitting test, etc., Celts of P. fluorescens well in medium containing 30g/L of glucose, whereas the growth was inhibited at the glucose levels at higher than 30g/L. The highest values of specific growth rate and productivity were obtained when using 10g/1 of yeast extract. Optimum concentrations of $NH_4Cl$ and ${(NH_4Cl)}_2SO_4$ for culture were found to be 1.0g/L and 0.1g/L respectively. Optimum concentration of $MgSO_4{\cdot}7H_2O$ used as a sulfursource was 1.0g/L. It was also found that the cell concentrations reached the maximum level when grown on the medium containing 1.0g/L of $KH_2PO_4$ and 0.1g/L of $CaCl_2$. Also, the optimum culture conditions were $30^{\circ}C$ and pH 6.0. Cultivation of P. fluarescens at high dissolved oxygen (DO) concentration led to a decrease of bacterial productivity in batch culture. Maximum productivity was achieved at 40% DO concentration.

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Bacterial growth and carbon-to-phosphorus consumption in drinking water with different carbon and phosphorus levels (수돗물의 탄소와 인 농도에 따른 세균의 생장과 C/P 소모율)

  • Choi, Sung-Chan;Park, e-Keun;Kim, Yeong-Kwan
    • Journal of Korean Society of Water and Wastewater
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    • v.27 no.6
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    • pp.689-701
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    • 2013
  • Bacterial growth and corresponding consumption of carbon and phosphorus were examined in which tap water samples containing a very low concentration of free chlorine were supplemented with organic carbon and/or phosphorus. The experiments were performed in a fed-batch mode under a controlled temperature of $20^{\circ}C$. In the phosphorus alone-added water, there was no significant increase in bacterial numbers measured as heterotrophic plate count (HPC) in the bulk water. However, bacterial growth was stimulated by the addition of carbon (e.g., bulk HPC levels increased to $10^3CFU/mL$) and further stimulated by the combined addition of carbon and phosphorus (e.g., bulk HPC to $10^5CFU/mL$). The same effects were observed in biofilm HPC and biomass formed on polyethylene (PE) slide surfaces. In the water where organic carbon and phosphorus were added together, the highest biofilm HPC and biomass (measured as extracellular polymeric substance components) densities were observed which were $7.6{\times}10^5CFU/cm^2$ and $5.3{\mu}g/cm^2$, respectively. In addition to the bacterial growth, additions of organic carbon and/or phosphorus resulted in different bacterial carbon-to-phosphorus (C/P) consumption ratios. Compared to a typical bacterial C/P consumption ratio of 100:1, a higher C/P ratio (590:1) occurred in the carbon alone-added water, while a lower ratio (40:1) in phosphorus alone-added water. Comparative value (80:1) of C/P ratio was also observed in the water where organic carbon and phosphorus were added together. At the given experimental conditions, bacterial growth was deemed to be more sensitive to microbially available organic carbon than phosphorus. The effect of phosphorus addition, which resulted in a lower C/P consumption ratio, seemed to be tightly associated with the presence of microbially available organic carbon. These results suggested that the control of extrinsic carbon influx seemed to be more important to minimize bacterial regrowth in drinking water system, since even low content of phosphorus naturally occurring in drinking water was enough to allow a bacterial growth.

Overexpression of Thermoalkalophilic Lipase from Bacillus stearothermophilus L1 in Saccharomyces cerevisiae

  • Ahn, Jung-Oh;Jang, Hyung-Wook;Lee, Hong-Weon;Choi, Eui-Sung;Haam, Seung-Joo;Oh, Tae-Kwang;Jung, Joon-Ki
    • Journal of Microbiology and Biotechnology
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    • v.13 no.3
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    • pp.451-456
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    • 2003
  • An expression vector system was developed for the secretory production of recombinant Bacillus stearothermophilus L1 lipase in Saccharomyces cerevisiae. The mature L1 lipase gene was fused to ${\alpha}-amylase$ signal sequence from Aspergillus oryzae for the effective secretion into the culture broth and the expression was controlled under GAL10 (the gene coding UDP-galactose epimerase of S. cerevisiae) promoter. S. cerevisiae harboring the resulting plasmid successfully secreted L1 lipase into the culture broth. To examine an optimum condition for L1 lipase expression in the fed-batch culture, L1 lipase expression was induced at three different growth phases (early, mid, and late-exponential growth phases). Maximum product on of L1 lipase (1,254,000 U/l, corresponding to 0.65/1) was found when the culture was induced at an early growth phase. Secreted recombinant L1 lipase was purified only through CM-Sepharose chromatography, and the purified enzyme showed 1,963 U/mg of specific activity and thermoalkalophilic properties similar to those reported for the enzyme expressed in Escherichia coli.

Two-Stage Fermentation for 2-Ketogluconic Acid Production by Klebsiella pneumoniae

  • Sun, Yuehong;Wei, Dong;Shi, Jiping;Mojovic, Ljiljana;Han, Zengsheng;Hao, Jian
    • Journal of Microbiology and Biotechnology
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    • v.24 no.6
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    • pp.781-787
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    • 2014
  • 2-Ketogluconic acid production by Klebsiella pneumoniae is a pH-dependent process, strictly proceeding under acidic conditions. Unfortunately, cell growth is inhibited by acidic conditions, resulting in low productivity of 2-ketogluconic acid. To overcome this deficiency, a two-stage fermentation strategy was exploited in the current study. During the first stage, the culture was maintained at neutral pH, favoring cell growth. During the second stage, the culture pH was switched to acidic conditions favoring 2-ketogluconic acid accumulation. Culture parameters, including switching time, dissolved oxygen levels, pH, and temperature were optimized for the fed-batch fermentation. Characteristics of glucose dehydrogenase and gluconate dehydrogenase were revealed in vitro, and the optimal pHs of the two enzymes coincided with the optimum culture pH. Under optimum conditions, a total of 186 g/l 2-ketogluconic acid was produced at 26 h, and the conversion ratio was 0.98 mol/mol. This fermentation strategy has successfully overcome the mismatch between optimum parameters required for cell growth and 2-ketogluconic acid accumulation, and this result has the highest productivity and conversion ratio of 2-ketogluconic and produced by microorganism.