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Fed-Batch Sorbose Fermentation Using Pulse and Multiple Feeding Strategies for Productivity Improvement

  • Giridhar, R.;Srivastava, A.K.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.5
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    • pp.340-344
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    • 2000
  • Microbial oxidation of D-sorbitol to L-sorbose by Acetobacter suboxydans is of commercial importance since it is the only biochemical process in vitamin C synthesis. The main bottleneck in the batch oxidation of sorbitol to sorbose is that the process is severely inhibited by sorbitol. Suitable fed-batch fermentation designs can eliminate the inherent substrate inhibition and improve sorbose productivity. Fed-batch sorbose fermentations were conducted by using two nutrient feeding strategies. For fed-batch fermentation with pulse feeding, highly concentrated sorbitor (600g/L) along with other nutrients were fed intermittently in four pulses of 0.5 liter in response to the increased DO signal. The fed-batch fermentation was over in 24h with a sorbose productivity of 13.40g/L/h and a final sorbose concentration of 320.48g/L. On the other hand, in fed-batch fermentation with multiple feeds, two pulse feeds of 0.5 liter nutrient medium containing 600g/L sorbitol was followed by the addition of 1.5 liter nutrient medium containing 600g/L sorbitol at a constant feed rate of 0.36L/h till the full working capacity of the reactor. The fermentation was completed in 24h with an enhanced sorbose productivity of 15.09g/L/h and a sorbose concentration of 332.60g/L. The sorbose concentration and productivity obtained by multiple feeding of nutrients was found to be higher than that obtained by pulse feeding and was therefore a better strategy for fed-batch sorbose fermentation.

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Characteristics of Disinfection By-Products Formation in Korea (국내 정수장의 소독부산물 생성 특성)

  • Kim, Jinkeun;Jeong, Sanggi;Shin, Changsoo;Cho, Hyukjin
    • Journal of Korean Society of Water and Wastewater
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    • v.19 no.3
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    • pp.301-311
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    • 2005
  • The characteristics of disinfection by-products (DBPs) formation at 28 water treatment plants in Korea were investigated. Investigated species of DBPs were trihalomethanes (THMs), haloacetic acids (HAAs) and chloral hydrate (CH). The maximum concentration of THMs was $84.1{\mu}g/L$, minimum and the averages were $6.9{\mu}g/L$ and $27.8{\mu}g/L$, respectively; the maximum concentration of $HAA_5$ was $90.8{\mu}g/L$, minimum and the averages were $3.8{\mu}g/L$ and $26.7{\mu}g/L$, respectively; while the maximum concentration of CH was $29.5{\mu}g/L$, minimum and the averages were $0.5{\mu}g/L$ and $7.4{\mu}g/L$, respectively. On the other hand, DBPs levels during summer months, when the water temperature was near $25^{\circ}C$, were nearly twice as great as DBPs levels during the winter season. The ratio of $THMs/HAA_5$ was 1.07, and $HAA_5$ and THMs were the dominant species of DBPS in the Kum-Sumjin river and Nakdong river, respectivley.

Optimization of Xylitol Production by Candida tropicalis in Two-stage Fed-batch Culture (Candida tropicalis의 2단계 유가식 배양에 의한 Xylitol 생산의 최적화)

  • 유연우;조영일;서진호
    • KSBB Journal
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    • v.17 no.1
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    • pp.93-99
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    • 2002
  • Two-stage fed-batch culture of Candide tropicalis that was designated primarily to cultivate the cell in the glucose medium (1st stage) and then produced the xylitol from xylose medium (2nd stage) was developed to improve a xylitol yield and productivity. In the growth stage, glucose was automatically supplied to the fermentor by pH-stat mode when the pH was up 5.7, When a feeding medium was added in order to reach the glucose and yeast extract concentrations up to 100 and 40 g/L, respectively, a high cell concentration and a relatively low ethanol concentration were obtained in 18.5 h culture. In the production stage, initial xylose concentration of 150 g/L was the most favorable for obtaining the final xylitol concentration and productivity. The addition of mineral salts was also enhanced a xylitol production. But the aeration rate was not significantly affected a xylitol production. When the addition of 16 g yeast extract and 232.5 g xylose powder at the production stage was used, xylitol yield and productivity were significantly increased. With these conditions, xylitol concentration, yield and productivity of 108.9 g/L, 74%) and 3.3 g/L·h, respectively, were obtained in a final volume of 1.58 L. The further addition of 16 g yeast extract and 232.5 g xylose powder increased the working volume partly (1.67 L) and resulted in a relatively high xylitol concentration, yield and productivity of 193 g/L, 70% and 3.6 g/L·h, respectively.

Enhancement of Ginsenoside Rg1 and Rg5 Contents in an Extract of Wood-cultivated Ginseng by Lactobacillus plantarum (Lactobacillus plantarum을 이용한 산양삼 추출물의 진세노사이드 Rg1 및 Rg5의 함량 증대)

  • Kwon, Hun-Joo;Cho, Yun-Ji;Kim, Myoung-Dong
    • Microbiology and Biotechnology Letters
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    • v.45 no.4
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    • pp.305-310
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    • 2017
  • Twelve lactic acid bacteria harboring ${\alpha}$-rhamnosidase (EC 3.2.1.40) activity were isolated from traditional Korean foods. The 6 strains (Weissella confuse [n = 1], Lactobacillus pentosus [n = 1], and Lactobacillus plantarum [n = 4]) with the highest rhamnosidase activity were selected for bioconversion of an extract of wood-cultivated ginseng. The L. plantarum MBE/L2990 strain increased ginsenoside content (0.58 mg for Rg1 and 0.24 mg for Rg5) and showed higher bioconversion activity than the control strain L. plantarum KCTC21004 (56% and 42% increase for Rg1 and Rg5, respectively). L. plantarum MBE/L2990 was deposited at the Korean Collection for Type Cultures as Lactobacillus plantarum KCTC18529P.

Comparing Compliance and Efficacy of Isocaloric Oral Nutritional Supplementation Using 1.5 kcal/mL or 1 kcal/mL Sip Feeds in Mildly to Moderately Malnourished Indonesian Children: A Randomized Controlled Trial

  • Devaera, Yoga;Syaharutsa, Danny Maesadatu;Jatmiko, Herwasto Kuncoroyakti;Sjarif, Damayanti Rusli
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.21 no.4
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    • pp.315-320
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    • 2018
  • Purpose: This study set out to evaluate the compliance to, and efficacy of oral supplementation, using a 1.5 kcal/mL or 1 kcal/mL sip feed, in children with mild to moderate malnutrition. Methods: This was a parallel, randomized, controlled open-label trial in children aged 3 to 6 years with a weight for height Z (WHZ) score <-1 and ${\geq}-3$, who were randomized to receive a total of 600 kcal/day from either a 1.5 kcal/mL or a 1.0 kcal/mL pediatric sip feed for 28 days. Assessments included daily study product intake, body weight, tolerance and dietary intake from solid food. Results: Of 110 children recruited, 98 ($mean{\pm}standard$ deviation of age $49{\pm}7months$) completed the study. Both sip feeds were well tolerated, with high compliance ($80{\pm}24%$ and $81{\pm}22%$ of prescribed volume in 1.5 kcal/mL and 1.0 kcal/mL groups respectively, p=0.79). Both study groups gained similar weight during the 28 days intervention period ($0.42{\pm}0.40kg$ in 1.5 kcal/mL group vs. $0.49{\pm}0.49kg$ in 1.0 kcal/mL group, p=0.43). There were no significant differences between the groups in weight gain and in the change in WHZ score over the intervention period. Dietary analysis at the end of the study did not show replacement of solid food by the oral nutritional supplements. Conclusion: In children with mild to moderate malnutrition, both 1.5 kcal/mL and 1 kcal/mL pediatric sip feeds had high compliance and were well tolerated, and were equally effective in promoting weight gain in the 28 days study period.

Analysis of Runoff Characteristics of NPS Pollution in different Slope from Upland field (경사도가 다른 고랭지 밭의 비점오염물질 유출특성 분석)

  • Lee, Su-In;Shin, Jae-Young;Shin, Min-Hwan;Choi, Yong-Hun;Choi, Joong-Dae
    • Proceedings of the Korea Water Resources Association Conference
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    • 2012.05a
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    • pp.906-906
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    • 2012
  • 밭에서 발생하는 비점오염물질은 강우량, 강우강도, 경사도, 토양, 시비량 등에 영향을 받는다. 따라서 효율적인 비점오염물질의 관리를 위해서는 영향인자에 대한 자세한 분석이 선행되어야 한다. 이에 본 연구에서는 강원도 평창군 대관령면의 경사도가 다른 고랭지 밭 두 지점을 선정하여 비점오염물질의 유출특성을 파악하고, 향후 비점오염원의 효율적인 관리를 위한 기초자료를 제공하고자 하였다. 연구기간은 2011년 4월부터 11월까지 유출이 발생한 8회의 강우사상에 대해 모니터링을 수행하였다. 연구결과 경사도 4.9% 밭에서의 유출율은 0.05~0.48이었으며, 16.8% 경사에서는 0.31~0.57로 나타났다. 유량과 채취된 수질 시료를 이용해 EMC를 산정한 결과 경사도 4.9%인 밭에서는 SS 762.1~2422.7 mg/L $BOD_5$ 4.5~14.9 mg/L, $COD_{Cr}$ 16.1~62.0 mg/L, $COD_{Mn}$ 7.5~43.2 mg/L, TN 9.602~21.021 mg/L, TP 2.544~6.763 mg/L의 범위로 나타났다. 반면 16.8%의 경사밭에서는 SS 116.0~2015.5 mg/L $BOD_5$ 6.0~9.5 mg/L, $COD_{Cr}$ 21.9~75.4 mg/L, $COD_{Mn}$ 4.3~33.1 mg/L, TN 10.937~46.295mg/L, TP 2.611~11.197 mg/L의 범위로 나타났다. 두 지점의 유출률과 EMC를 비교한 결과, 경사가 증가함에 따라 유출율이 증가하는 것으로 나타났으며, SS를 제외한 모든 수질항목의 EMC가 증가한 것으로 나타났다. SS의 경우 경사가 큰 고랭지 밭에서 낮은 값을 보였는데, 이는 16.8%의 경사밭에서 등고선 방향으로 경운을 하여 흙 입자의 유실이 적었기 때문인 것으로 판단된다. 본 연구의 결과와 같이 밭에서 발생하는 비점오염물질의 유출특성은 강우조건(강우강도, 선행무강우일수), 시비량, 토양, 지표의 피복상태 등에 따라 다르기 때문에 장기적인 모니터링을 통해 기초자료를 축적하는 것이 중요하다고 판단된다.

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Inhibition of Listeria monocytogenes by Low Concentrations of Ethanol (저농도의 Ethanol에 의한 Listeria monocytogenes의 증식억제)

  • 박찬성;김미림
    • Korean journal of food and cookery science
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    • v.11 no.4
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    • pp.379-385
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    • 1995
  • The effect of low concentrations of ethanol (3-7%, v/v) in tryptic soy broth (TSB) as an antibacterial agent against Listeria monocytogenes was tested at -20, 5, 35, 45, 50 and 55$^{\circ}C$. Increasing concentrations of ethanol progressively inhibited initial growth of L. monocytogenes at 35$^{\circ}C$. Growth occured at 5% ethanol, but only after a prolonged lag period. The number of viable cells of L. monocytogenes declined during incubation at 7% ethanol. TSB containing 3-7% ethanol was inoculated with 10$\^$5/-10$\^$6/ cells/$m\ell$ or L. monocytogenes and incubated at low temperatures (5$^{\circ}C$, -20$^{\circ}C$). In the presence of 3% of ethanol at 5 or -20$^{\circ}C$, bacterial growth was inhibited more than 90% of control cells. TSB containing 3-7% ethanol was inoculated with 10$\^$6/-10$\^$7/ cells/$m\ell$ of L. monocytogenes and incubated at high temperatures (45$^{\circ}C$, 50$^{\circ}C$, 55$^{\circ}C$). Decrease in viability of the cells incubated at 45 or 50$^{\circ}C$ was slow and the survival of L. monocytogenes was not affected so much in the presence of 3% of ethanol. The viability of L. monocytogenes was decreased with increasing concentration of ethanol and temperature. Decimal reduction times (D-values) based on tryptic soy agar plates at 55$^{\circ}C$ were 20.1, 12.6, 7.4 and 4.2 min in 0, 3, 5 and 7% ethanol, respectively.

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Isolation, Identification and Mutant Development of Butanol Tolerance Bacterium (부탄올 내성 미생물의 분리, 동정 및 변이주의 개발)

  • Jung, Hyesook;Lee, Jinho
    • Microbiology and Biotechnology Letters
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    • v.41 no.1
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    • pp.26-32
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    • 2013
  • Butanol-resistant bacteria were isolated from butanol solvent. The cell growth of isolated strains declined with increasing concentrations of butanol, and isolated strain BRS02 displayed more resistance to 12.5 g/L of butanol than other isolated strains. In addition, strain BRS251, which was resistant to even higher concentrations of butanol, was developed by the mutation of BRS02 using UV. BRS251 could grow in LB medium containing up to 17.5 g/L of butanol, 32.5 g/L of propanol, or 6 g/L of pentanol, whereas the control strain Escherichia coli was found to be tolerant to 7.5 g/L of butanol, 20 g/L of propanol, or 2 g/L of pentanol. The isolated BRS02, a Gram(+) bacterium seen to have a cocci form under the microscope, grew in 6.5% NaCl. According to biochemical tests, BRS02 can metabolize and produce acid with D-galactose, D-maltose, D-mannitol, D-mannose, methyl-${\beta}$-Dglucopyranoside, D-ribose, sucrose, or D-trehalose, as carbon sources. Also, this strain showed resistance to bacitracin, vibriostatic agent O/129, and optochin, alongside positive activities for arginine dihydrolase, ${\alpha}$-glucosidase, and urease. The BRS02 strain was identified as Staphylococcus sp. by analyses of the 16S rRNA gene, phylogenetic tree, and biochemical tests.

Induction and in vitro Proliferation of Adventitious Roots in Phyllanthus urinaria (여우구슬(Phyllanthus urinaria)의 부정근 유도 및 기내증식조건)

  • Bae, Kee-Hwa;Yun, Pil-Yong;Choi, Yong-Eui
    • Korean Journal of Plant Resources
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    • v.22 no.5
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    • pp.454-460
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    • 2009
  • Phyllanthus urinaria was an important species in Korea and distributed in all around of Korea. The roots and stems of this plant have been used for natural medicine for the treatment of diabetes, the hepatitis B virus and disturbances of the kidney and urinary bladder. Production of adventitious roots in P. urinaria by in vitro cultures could be used as alternatives materials. Shoot and root segments from P. urinaria seedling were cultured on Murashige and Skoog (MS) medium supplemented with 3.0 mg/L IBA and 30 g/L sucrose. After 4 weeks of culture, the highest induction of adventitious roots was obtained from the shoot part. Frequency of adventitious root formation on medium with various kinds of auxins (IAA, NAA, 2,4-D, and IBA) and various concentrations of IBA (0, 0.1, 0.5, 1.0, 3.0, and 5.0 mg/L) was tested. The maximun induction of adventitious root was obtained on medium with 0.5 mg/L IBA. In liquid culture, growth of root was best on medium supplemented with 30 g/L sucrose. Adventitious roots were cultured in 5 L bioreactor containing 1/2 MS medium supplemented with 0.5 mg/L IBA and 30 g/L sucrose and mass-production of adventitious roots was successfully achieved. These results revealed the first attempt for the production of adventitious roots in P. urinaria.

젖산균의 Plasmid DNA 분리방법 및 Electroporation에 의한 젖산균의 형질전환에 관한 연구

  • Kim, Seon-Gi
    • 한국유가공학회:학술대회논문집
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    • 1997.05a
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    • pp.41-61
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    • 1997
  • 젖산균의 유전자 연구를 촉진하기 위해 간단하고 신속한 plasmid DNA의 분리방법과 electro-poration을 이용하여 vector plasmid의 간단하고 신속한 전이방법을 얻기 위해 젖산균의 형질전환에 영향하는 요인에 대하여 연구하였으며 연구결과는 다음과 같다. 1. O'Sullivan과 Klaenhammer의 방법을 개선하여 젖산균 plasmid DNA의 분리에 좋은 결과를 얻을 수 있는 신속하고 쉬운 방법을 고안하였으며, genomic DNA 분리에 이용되는 guanidium thio-cyanate 처리방법을 plasmid의 분리에 적용할 수 있었다. 2. L. casei, L. acidophilus. L. delbruekii var. bulgaricus. L. brevis와 L. plantarum 균주에서 plasmid를 확인하였으며, 돼지 분에서 분리된 L. lactis ssp. lactis. L. fermentum과 L. plantarum에서도 plasmid를 분리 확인하였다. 3. Lactococci의 plasmid분리는 lactobacilli와는 달리 mutanolysin의 처리없이도 잘 되었으며, L. lactis ssp. lactis와 Ent. faecalis에서 plasmid를 확인하였다. 4. E. coli plasimd 분리에 이용되는 MPS membrane filter 방법으로 젖산균 plasmid pLZ12의 분리가 가능하였으나, 세포파편이 filter를 막아 사용에 어려움이 있는 것으로 확인되었다. 5. Plasmid 분리없이 electroporation을 이용한 세포 대 세포 전이법으로 간편하고 빠르게 E. coli DH5${\alpha}$에 E. coli Jm109의 plasmid pBX19, pBR322를 전이시켰다. 6. L. lactis ssp. lactis 균주에 lysozyme 처리시 30${\sim}$80%의 생존율을 보였으며, 대부분의 L. acidophilus 균주의 경우 약 70%의 생존율을 보였다. L. casei 102S의 경우는 45분간 처리 시에도 100%의 생존율을 보였다. 8. L. lactis ssp. lactis 균주에 pLZ12를 6.0kV에서 전이시킨 결과 12.5kV에서보다 형질전환 효율이 훨씬 높았으며 lysozyme 처리에 의해 형질전환 효율이 증가되었다. 9. L. acidophilus 균주에 pLZ12를 전이시 6.0kV에서는 전이가 모두 이루어졌으나, 12.5kV에서는 L. acidophilus WIESBY와 NCFM에서 전이가 이루어지지 않았으며, lysozyme 처리 후 pLZ12를 전이시켰을 때 12kV보다 6.0kV에서 형질전환 효율이 증가되었다. 10. Gene Pulser와 Progenitor II를 사용하여 pLZ12를 L. lactis ssp. lactis 균주에 전이하였을 때 Gene Pulser에 비해 Progenitor II의 형질전환 효율이 현저히 떨어졌다. L. acidophilus HY7008과 HY7001은 두 기기 모두 형질전환이 이루어졌으나, L. acidophilus WEISBY와 NCFM은 Progeni-tor II에서 전이가 일어나지 않았으며, Gene Pulser에서 전이균주를 얻어 두 electroporator간에 형질전환 효율의 차이를 보였다. 11. L. casei 102S에 pLZ12를 electroporation시 낮은 전압에서 형질전환 효율이 비교적 좋았으며, 배양 시기를 달리하여 전이시켰을 때 대수생장 말기의 세포가 형질전환 효율이 좋았다. 12. L. casei 102S세포를 각각 10% glycerol, EB, 2차 증류수 등에 녹여 electroporation을 실시하였을 때 각각 $3.8{\times}10^3$, $5.0{\times}10^2$,1.5${\times}10^2$cfu의 형질전환 효율을 보였으며, 1.0mM HEPES, TE buffer를 사용하였을 때에는 전이가 이루어지지 않았다. 13. Plasmid pLZ12의 농도를 달리하여 electroporation을 하였을 때 형질전환 효율이 농도에 비례하여 증가하였다. 14. L. casei 102S에 대수생장 말기의 세포를 채취하여 10% glycerol, 200 Ohms, 25 ${\mu}$FD, 10kV/cm로 plasmid pLZ12를 electroporation할 때 최대 형질전환 효율인 3.8${\times}$10$^{3}$cfu를 얻었으며, lysozyme 처리가 다른 젖산균과는 달리 형질전환 효율을 증가시키지 못하였다. 15. L. casei 102S 세포를 10% glycerol과 EB에 녹여 -20$^{\circ}C$에서 냉동시킨 다음 1일과 7일 후의 세포를 electroporation한 결과 냉동시 세포에 손상을 주는 것으로 인식되었다.

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